IP2007 Vol 2 (2) +

INDIAN
PHARMACOPOEIA
2007
Volume 2
THE INDIAN PHARMACOPOEIA COMMISSION

GHAZIABAD
INDIAN PHARMACOPOEIA 2007
Volume 2
CONTENTS
General Monographs on Dosage Forms .................................................................
Monographs on Drug Substances, Dosage Forms and Pharmaceutical Aids

Monographs A to .............................................................................................
INDIAN PHARMACOPOEIA 2007 GENERAL NOTICES
GENERAL NOTICES
General Statements ....

Name ....
Official and Official Articles ....
Official Standards ....
Added Substances ....
Alternative Methods ....
Meanings of Terms ....
Provisions Applicable to Monographs and Test Methods ....
Expression of Contents ....
Expression of Concentrations ....
Abbreviated Statements ....
Weights and Measures ....
Monographs ....
General Monographs ....
Production ....
Manufacture of Drug Products ....
Excipients ....
Individual Monographs ....
Titles ....
Chemical Formulae ....
Atomic and Molecular Weights ....
Definitions ....
Statement of Contents ....
Descriptions ....
Identification ....
Tests and Assay ....
Tests ....
Other tests ....
Limits ....
Quantities ....
5
GENERAL NOTICES INDIAN PHARMACOPOEIA 2007
Apparatus ....
Reagents and Solutions ....
Indicators ....
Reference Substances ....
Tests Animals ....
Calculation of Results ....
Storage ....
Storage Containers ....
Labelling ....
6
IP 2007 GENERAL NOTICES
General Notices use but not necessarily to articles that may be sold under the
same name for other purposes.
The active pharmaceutical ingredients (drug substances),
General Statements excipients (pharmaceutical aids), pharmaceutical preparations
The General Notices provide the basic guidelines for the (dosage forms) and other articles described in the monographs
interpretation and application of the standards, tests, assays, are intended for human and veterinary use (unless explicitly
and other specifications of the Indian Pharmacopoeia (IP), as restricted to one of these uses).
well as to the statements made in the monographs and other The requirements given in the monographs are not framed to
texts of the Pharmacopoeia. provide against all possible impurities, contaminants or
A monograph is to be constructed in accordance with any adulterants; they provide appropriate limitation of potential
general monograph or notice or any appendix, note or other impurities only.
explanatory material that is contained in this Pharmacopoeia A preparation must comply throughout the shelf-life assigned
and that is applicable to that monograph. All statements to it by the manufacturer; for opened or broached containers
contained in the monograph, except where a specific general the maximum period of validity for use may sometimes be
notice indicates otherwise and with the exceptions given stated in the individual monograph. Nevertheless, the
hereafter, constitute standards for the official articles. An article responsibility for assigning the period of validity shall be
is not of pharmacopoeial quality unless it complies with all of with the manufacturer.
the requirements stated. Added Substances. An official substance, as distinguished
Exceptions to the General Notices do exist, and where they from an official preparation, contains no added substances
do, the wording in the individual monograph or an appendix except when specifically permitted in the individual monograph.
takes precedence and specifically indicates directions or the Unless otherwise specified in the individual monograph, or
intent. Thus, the specific wording of standards, tests, assays elsewhere in the General Notices, suitable substances may be
and other specifications is binding wherever deviations from added to an official preparation to enhance its stability,
the General Notices exist. Likewise, where there is no specific usefulness or elegance, or to facilitate its preparation. Such
mention to the contrary, the General Notices apply. auxiliary substances shall be harmless in the amounts used,
shall not exceed the minimum quantity required to provide
Name. The full name or title of this book, including addenda
their intended effect, shall not impair the therapeutic efficacy
thereto, is Indian Pharmacopoeia 2007, abbreviated to IP 2007.
or the bioavailability or safety of the preparation and shall not
In the texts, the term “Pharmacopoeia” or “IP” without
interfere with the tests and assays prescribed for determining
qualification means the Indian Pharmacopoeia 2007 and any
compliance with the official standards. Particular care should
addenda thereto.
be taken to ensure that such substances are free from harmful
Official and Official Articles. The word ‘official’ wherever organisms. The freedom to the manufacturers to add auxiliary
used in this Pharmacopoeia or with reference thereto, is substances imposes on them the responsibility of satisfying
synonymous with ‘pharmacopoeial’, with ‘IP’ and with the licensing authorities on the purpose of the addition and
‘compendial’. The designation IP in conjunction with the the innocuity of such substances.
official title on the label of an article is an indication that the Alternative Methods. The tests and assays described are the
article purports to comply with IP standards. official methods upon which the standards of the
The following terms are used where the articles for which Pharmacopoeia are based. Alternative methods of analysis
monographs are provided are to be distinguished. may be used for control purposes, provided that the methods
used are shown to give results of equivalent accuracy and
An official substance is a single drug or a drug entity or a enable an unequivocal decision to be made as to whether
pharmaceutical aid for which the monograph title includes no compliance with the standards of the monographs would be
indication of the nature of a dosage form. achieved if the official methods were used. Automated
An official preparation is a drug product (dosage form) and is procedures utilising the same basic chemistry as the test
the finished or partially finished preparation or product of one procedures given in the monograph may also be used to
or more official substances formulated for use on the patient. determine compliance. Such alternative or automated
procedures must be validated.
An article is an item for which a monograph is provided,
whether an official substance or an official preparation. In the event of doubt or dispute, the methods of analysis of
the Pharmacopoeia are alone authoritative and only the result
Official Standards. The requirements stated in the obtained by the procedure given in this Pharmacopoeia is
monographs apply to articles that are intended for medicinal conclusive.
7
GENERAL NOTICES IP 2007
Meanings of Terms — per cent v/v (percentage, volume in volume) expressing

Alcohol. The term “alcohol” without qualification means the number of millilitres of substance in 100 millilitres of
ethanol (95 per cent). Other dilutions of ethanol are indicated final product.
by the term “alcohol” or “alcohol” followed by a statement of The expression “parts per million” refers to the weight in
the percentage by volume of ethanol (C2H6O) required. weight, unless otherwise stated.
Desiccator. A tightly-closed container of suitable size and Where the content of a substance is expressed in terms of the
design that maintains an atmosphere of low moisture content chemical formula for that substance an upper limit exceeding
by means of silica gel or phosphorus pentoxide or other 100 per cent may be stated. Such an upper limit applies to the
suitable desiccant. result of the assay calculated in terms of the equivalent content
of the specified chemical formula. For example, the statement
Drying and ignition to constant weight. Two consecutive
‘contains not less than 99.0 per cent and not more than 101.0
weighings after the drying or igniting operations do not differ
per cent of C7H6O2 implies that the result of the assay is not
by more than 0.5 mg, the second weighing following an
less than 99.0 per cent and not more than 101.0 per cent,
additional period of drying or of ignition respectively
calculated in terms of the equivalent content of C7H6O2.
appropriate to the nature and quantity of the residue.
Where the result of an assay or test is required to be calculated
Ethanol. The term “ethanol” without qualification means
with reference to the dried, anhydrous, ignited substance, or
anhydrous ethanol or absolute alcohol.
the substance free from solvent, the determination of loss on
Filtration. Unless otherwise stated, filtration is the passing of drying, water content, loss on ignition, content of the specified
a liquid through a suitable filter paper or equivalent device solvent, respectively is carried out by the method prescribed
until the filtrate is clear. in the relevant test in the monograph.
Freshly prepared. Made not more than 24 hours before it is Expression of Concentrations. The following expressions in
issued for use. addition to the ones given under Expression of Content are
also used:
Label. Any printed packing material, including package inserts
that provide information on the article. — per cent w/v (percentage, weight in volume) expressing
the number of grams of substance in 100 millilitres of
Negligible. A quantity not exceeding 0.50 mg. product
Solution. Where the name of the solvent is not stated, — per cent v/w (percentage, volume in weight) expressing
“solution” implies a solution in water. The water used complies the number of millilitres of substance in 100 grams of
with the requirements of the monograph on Purified Water. product.
The term ‘distilled water’ indicates Purified Water prepared by
distillation. Usually, the strength of solutions of solids in liquids is
expressed as percentage weight in volume, of liquids in liquids
Temperature. The symbol º used without qualification as percentage volume in volume, of solids in semi-solid bases
indicates the use of the Celsius thermometric scale. (e.g. creams) and of gases in liquids as percentage weight in
Water. If the term is used without qualification it means Purified weight.
Water of the Pharmacopoeia. The term ‘distilled water’ When the concentration of a solution is expressed as parts of
indicates Purified Water prepared by distillation. dissolved substance in parts of solution, it means parts by
weight (g) of a solid in parts by volume (ml) of the final solution;
Water-bath. A bath of boiling water unless water at another
as parts by weight (g) of a gas in parts by weight (g) of the
temperature is indicated. Other methods of heating may be
final solution.
used provided the required temperature is approximately
maintained but not exceeded. When the concentration of a solution is expressed in molarity
designated by the symbol M preceded by a number, it denotes
Provisions Applicable To Monographs and Test Methods the number of moles of the stated solute contained in sufficient
Purified Water (unless otherwise stated) to produce 1 litre of
Expression of Content. Where the content of a substance is solution.
defined, the expression “per cent” is used according to
circumstances with one of two meanings: Abbreviated Statements. Incomplete sentences are employed
in parts of the monographs for directness and brevity (for
— per cent w/w (percentage, weight in weight) expressing example, Iodine Value. Not more than ……; Relative Density.
the number of grams of substance in 100 grams of final …….to……..) Where the tests are abbreviated, it is to be
product, understood that the test method referred to in brackets
8
provides the method to be followed and that the values Excipients. Any substance added in preparing an official
specified are the applicable limits. preparation shall be innocuous, shall have no adverse influence
in the therapeutic efficacy of the active ingredients and shall
Weights and Measures. The metric system of weights and
not interfere with the tests and assays of the Pharmacopoeia.
measures is employed in the Pharmacopoeia. All measures are
Care should be taken to ensure that such substances are free
required to be graduated at 25º and all measurements in tests
from harmful organisms.
and assays, unless otherwise stated, are to be made at that
temperature. Graduated glass apparatus used in analytical Individual Monographs
operations shall comply with the requirements stated in
Chapter 2.1.6 Drug products that are the subject of an individual monograph
are also required to comply with the tests given in the general
monographs.
Monographs
Titles. The main title for a drug substance is the International
General Monographs Non-proprietary Name (INN) approved by the World Health
Organization. Subsidiary names and synonyms have also been
General monographs on dosage forms include requirements given in some cases; where included, they have the same
of general application and apply to all preparations within the significance as the main title.
scope of the Introduction section of the general monograph,
except where a preamble limits the application. The The main titles of drug products are the ones commonly
requirements are not necessarily comprehensive for a given recognised in practice. Synonyms drawn from the full non-
specific preparation; additional requirements may sometimes proprietary name of the active ingredient or ingredients have
be given in the individual monograph for it. also been given. Where, however, a product contains one or
the other of different salts of an active molecule, the main title
Production. Statements given under the heading Production is based on the full name of the active ingredient. For example,
relate to particular aspects of the manufacturing process and Chloroquine Phosphate Tablets and Chloroquine
are not necessarily comprehensive. However, they are SulphateTablets.
mandatory instructions to manufacturers. They may relate,
for example, to source materials, to the manufacturing process Chemical Formulae. When the chemical structure of an official
and its validation and control, to any in-process testing that substance is known or generally accepted, the graphic and
is to be carried out by the manufacturer on the final product molecular formulae are normally given at the beginning of the
either on selected batches or on each batch prior to release. monograph for information. This information refers to the
All this cannot be verified on a sample of the final product by chemically pure substance and is not to be regarded as an
an independent analyst. It is for the licensing authority to indication of the purity of the official material. Elsewhere, in
verify that the instructions have been followed. statement of purity and strength and in descriptions of
processes of assay, it will be evident from the context that the
The absence of a section on Production does not imply that formulae denote the chemically pure substances.
attention to features such as those given above is not required.
An article described in a monograph of the Pharmacopoeia is Where the absolute stereochemical configuration is specified,
to be manufactured in accordance with the principles of good the International Union of Pure and Applied Chemistry
manufacturing practice and in accordance with the (IUPAC) R/S and E/Z systems of designation have been used.
requirements of the Drugs and Cosmetics Rules, 1945. The If the substance is an enantiomer of unknown absolute
general principles applicable to the manufacture and quality stereochemistry, the sign of the optical rotation, as determined
assurance of drugs and preparations meant for human use in the solvent and under the conditions specified in the
apply equally to veterinary products as well. monograph, has been attached to the systematic name. An
indication of sign of rotation has also been given where this is
Manufacture of Drug Products. The opening definitive incorporated in a trivial name that appears on an IUPAC
statement in certain monographs for drug products is given in preferred list.
terms of the active ingredient(s) only. Any ingredient(s) other
than those included in the statement, must comply with the Atomic and Molecular Weights. The atomic weight or
general notice on Excipients and the product must conform to molecular weight is shown , as and when appropriate at the
the Pharmacopoeial requirements. top right hand corner of the monograph. The atomic and
molecular weights and graphic formulae do not constitute
Official preparations are prepared only from ingredients that analytical standards for the substances described.
comply with the requirements of the pharmacopoeial
monographs for those individual ingredients for which Definition. The opening statement of a monograph is one
monographs are provided. that constitutes an official definition of the substance,
9
preparation or other article that is the subject of the are not framed to take into account all possible impurities. It is
monograph. In certain monographs for pharmaceutical not to be presumed, for example, that an impurity that is not
preparations the statement is given in terms of the principal detectable by means of the prescribed tests is tolerated.
ingredient(s). Material found to contain such an impurity is not of
In monographs on vegetable drugs, the definition indicates pharmacopoeial quality if the nature or amount of the impurity
whether the subject of the monograph is, for example, the found is incompatible with good pharmaceutical practice.
whole drug or the drug in powdered form. Pharmacopoeial methods and limits should be used merely as
Certain pharmaceutical substances and other articles are compliance requirements and not as requirements to guarantee
defined by reference to a particular method of manufacture. A total quality assurance. Tests and assays are prescribed for
statement that a substance or article is prepared or obtained the minimum sample available on which the attributes of the
by a certain method constitutes part of the official definition article should be measured. Assurance of quality must be
and implies that other methods are not permitted. A statement ensured by the manufacturer by the use of statistically valid
that a substance may be prepared or obtained by a certain sampling and testing programmes.
method, however, indicates that this is one possible method Tests. Unless otherwise stated, the assays and tests are carried
and does not imply that other methods are not permissible. out at a temperature between 20º and 30º.
Statement of content. The limits of content stated are those Where it is directed that an analytical operation is to be carried
determined by the method described under Assay. out ‘in subdued light’, precautions should be taken to avoid
Description. The statements under the heading Description exposure to direct sunlight or other strong light. Where a
are not to be interpreted in a strict sense and are not to be procedure is directed to be performed ‘protected from light’
regarded as official requirements. precautions should be taken to exclude actinic light by the
Solubility. Statements on solubility are given in Chapter 2.4.26 use of low-actinic glassware, working in a dark room or similar
and are intended as information on the approximate solubility procedures.
at a temperature between 15º and 30º, unless otherwise stated, For preparations other than those of fixed strength, the
and are not to be considered as official requirements. However, quantity to be taken for a test or an assay is usually expressed
a test for solubility stated in a monograph constitutes part of in terms of the active ingredient. This means that the quantity
the standards for the substance that is the subject of that of the active ingredient expected to be present and the quantity
monograph. of the preparation to be taken are calculated from the strength
stated on the label.
Test Methods
Other Tests. In the monographs on dosage forms and certain
References to general methods of testing are indicated by test preparations, under the sub-heading ‘Other tests’ it is stated
method numbers in brackets immediately after the heading of that the article complies with the tests stated under the general
the test or at the end of the text. monograph of the relevant dosage form or preparation. Details
Identification. The tests given under the heading Identification of such tests are provided in the general monographs.
are not necessarily sufficient to establish absolute proof of Limits. The limits given are based on data obtained in normal
identity. They provide a means of verifying that the identity analytical practice. They take into account normal analytical
of the material under examination is in accordance with the errors, of acceptable variations in manufacture and of
label on the container. deterioration to an extent that is acceptable. No further
In certain monographs alternative series of identification tests tolerances are to be applied to the limits for determining whether
are given; compliance with either one or the other set of tests or not the article under examination complies with the
is adequate to verify the identity of the article. requirements of the monograph.
When tests for infrared absorption are applied to material Quantities. Unless otherwise stated, the quantities to be taken
extracted from formulated preparations, strict concordance for assays, limit tests and other tests are of the substance
with the specified reference spectrum may not always be under examination.
possible, but nevertheless a close resemblance between the In tests with numerical limits and assays, the quantity stated
spectrum of the extracted material and the specified reference to be taken for testing is approximate. The amount actually
spectrum should be achieved. used, which may deviate by not more than 10 per cent from
that stated, is accurately weighed or measured and the result
Tests and Assays
of analysis is calculated from this exact quantity. In tests where
The tests and assays are the official methods upon which the the limit is not numerical but usually depends upon
standards of the Pharmacopoeia depend. The requirements comparison with the behaviour of a reference in the same
10
conditions, the stated quantity is taken for testing. Reagents Indian Pharmacopoeia Commission (IPC). They are the official
are used in the prescribed amounts. standards to be used in cases of arbitration. Secondary
Quantities are weighed or measured with an accuracy Standards (Working Standards) may be used for routine
commensurate with the indicated degree of precision. For analysis, provided they are standardized at regular intervals
weighings, the precision is plus or minus 5 units after the last against the Reference Substances
figure stated. For example, 0.25 g is to be interpreted as 0.245 Biological Reference Substances, also abbreviated to IPRS
g to 0.255 g. For the measurement of volumes, if the figure and Standard Preparations of antibiotics are issued by
after the decimal point is a zero or ends in a zero, e.g. 10.0 ml 0r agencies authorised by the IPC. They are standardized against
0.50 ml, the volume is measured using a pipette, a volumetric the International Standards and Reference Preparations
flask or a burette, as appropriate; in other cases, a graduated established by the World Health Organization (WHO). The
measuring cylinder or a graduated pipette may be used. potency of these preparations is expressed in International
Volumes stated in microlitres are measured using a micropipette Units.
or microsyringe.
Reference spectra are published by the IPC and they are
The term ‘transfer’ is used generally to indicate a quantitative accompanied by information concerning the conditions used
operation. for sample preparation and recording of the spectra.
Apparatus. Measuring and weighing devices and other Test animals. Unless otherwise directed, animals used in a
apparatus are described in the chapter entitled ‘Apparatus for test or an assay shall be healthy and are drawn from a uniform
Tests and Assays’. A specification for a definite size or type stock, and have not previously been treated with any material
of container or apparatus in a test or assay is given merely as that will interfere with the test or the assay.
a recommendation. Calculation of results. In determining compliance with a
Unless otherwise stated, comparative tests are carried out numerical limit in assay or test, the result should be calculated
using identical tubes of colourless, transparent, neutral glass to one decimal place more than the significant figures stated
with a flat base, commonly known as Nessler cylinders. and then rounded up or down as follows: if the last figure
calculated is 5 to 9, the preceding figure is increased by 1; if it
Reagents and Solutions. The reagents required for the tests
is 4 or less, the preceding figure is left unchanged.
and assays of the Pharmacopoeia are defined in the various
chapters showing their nature, degree of purity and the Storage. Statements under the side-heading Storage constitute
strengths of the solutions to be made from them. The non-mandatory advice. The articles of the Pharmacopoeia are
requirements set out are not intended to imply that the materials to be stored under conditions that prevent contamination and,
are suitable for use in medicine; regents not covered by as far as possible, deterioration. Precautions that should be
monographs in the pharmacopoeia shall not be claimed to be taken in relation to the effects of the atmosphere, moisture,
of IP quality. heat and light are indicated, where appropriate, in the individual
monograph.
The term ‘analytical reagent grade of commerce’ implies that
the chemical is of a high degree of purity wherein the limits of Specific directions are given in some monographs with respect
various impurities are known. Where it is directed to use a to the temperatures at which Pharmacopoeial articles should
‘general laboratory reagent grade of commerce’ it is intended be stored, where it is considered that usage at a lower or
that a chemically pure grade material, not necessarily required higher temperature may produce undesirable results. The
to be tested for limiting or absence of certain impurities, is to storage conditions are defined by the following terms:
be used. — Store in a dry, well-ventilated place at a temperature not
Indicators. Where the use of an indicator solution is mentioned exceeding 30º
in an assay or test, approximately 0.1 ml of the solution shall — Store in a refrigerator (2º to 8º). Do not freeze
be added, unless otherwise directed.
— Store in a freezer (-2º to -18º)
Reference Substances. Certain monographs require the use — Store in a deep freezer (Below -18º)
of a chemical reference substance or a biological reference
preparation or a reference spectrum These are authentic Storage conditions not related to temperature are indicated in
specimens chosen and verified on the basis of their suitability the following terms:
for intended use as prescribed in the Pharmacopoeia and are — Store protected from light
not necessarily suitable in other circumstances. — Store protected from light and moisture
IP Reference Substances, abbreviated to IPRS (and referred Where no specific storage directions or limitations are given
to as RS in the individual monographs) are issued by the in the monograph or by the manufacturer, it is to be understood
11
that the storage conditions include protection from moisture, of being tightly closed, and re-closed after use.
freezing and excessive heat (any temperature above 40º).
In certain cases, special requirements of pack have been
Storage Containers. The requirements, guidance and indicated in some monographs under Storage, using
information on containers for pharmaceutical use are given in expressions that have been defined in chapter 6.1.
the chapter entitled Containers (6.1) Labelling. The labelling of drugs and pharmaceuticals is
In general, an article should be packed in a well-closed governed by the Drugs and Cosmetics Rules, 1945. The
container i.e. one that protects the contents from statements that are given in the monographs under the side-
contamination by extraneous solids, liquids or vapours and heading ‘Labelling’ are not comprehensive. Only those that
from loss of the article under normal conditions of handling are necessary to demonstrate compliance or otherwise with
and storage. the monograph have been given and they are mandatory. For
example, in the monograph on Betamethasone Sodium Tablets
Where, additionally, loss or deterioration of the article from
the labelling statement is “The label states the strength in
effervescence, deliquescence or evaporation under normal
terms of the equivalent amount of betamethasone”. Any other
conditions of storage is likely, the container must be capable
statements are included as recommendations.
12
INDIAN PHARMACOPOEIA 2007 GENERAL MONOGRAPHS
DOSAGE FORMS
General requirements ....

Capsules ....
Cream ....
Ear Drops ....
Eye Drops ....
Eye Ointments ....
Gels ....
Inhalation Preparations ....
Insulin Preparations ....
Nasal Preparations ....
Ointments ....
Oral Liquids ....
Oral Powders ....
Parenteral Preparations ....
Pessaries ....
Suppositories ....
Tablets ....
13
IP 2007 CAPSULES
General requirements Soft Gelatin Capsules. Soft gelatin capsules made from gelatin
(sometimes called softgels) or other suitable material require
The Pharmacopoeia provides monographs of dosage forms large-scale production methods. The soft gelatin shell is
for most of the pharmacopoeial drug substances. Additionally, somewhat thicker than that of hard-shell capsules and may be
the general requirements including the processes for the plasticized by the addition of a polyol such as sorbitol or
preparation of many of them and the tests of a general nature glycerin. The ratio of dry plasticizer to dry gelatin determines
applicable to each type of dosage form are given in the the “hardness” of the shell and may be varied to accommodate
following pages. In addition to defining the dosage forms, environmental conditions as well as the nature of the contents.
this section presents the general principles involved in the Like hard shells, the shell composition may include approved
production of some of them. dyes and pigments, opaquing agents such as titanium dioxide,
The requirement for compliance with the tests given under and preservatives. Flavors may be added and up to 5 per cent
each dosage form is indicated in each monograph of a drug sucrose may be included for its sweetness and to produce a
product under the heading ‘Other tests’. These tests are chewable shell. Soft gelatin shells normally contain 6 per cent
mandatory and are additional to the tests given in the individual to 13 per cent of water.
monograph. Soft gelatin capsules shells are usually formed, filled with
medicament and sealed in a combined operation on machines.
In some cases, shells for extemporaneous use may be
Capsules performed. The shells which are thicker than those of hard
capsules are formed to produce capsules which are spherical,
Capsules are solid dosage forms in which the drug or a mixture oval or cylindrical with hemispherical ends.
of drugs is enclosed in Hard Gelatin Capsule Shells, in soft,
Soft gelatin capsules also may be manufactured in a bubble
soluble shells of gelatin, or in hard or soft shells of any other
process that forms seamless spherical capsules. The shells
suitable material, of various shapes and capacities. They
may sometimes contain a medicament. They may contain a
usually contain a single dose of active ingredient(s) and are
preservative to prevent growth of fungi.
intended for oral administration. The consistency of soft shells
may be adjusted by the addition of substances such as The contents of soft capsules usually consist of liquids or
Glycerin and Sorbitol. Excipients such as opaque fillers, anti- solids dissolved or dispersed in suitable excipients to give a
microbial preservatives, sweetening agents, flavouring agents paste-like consistency. With suitable equipment, powders,
and one or more colouring agents permitted under the Drugs granules and other dry solids also may be filled into soft-shell
and Cosmetic Rules, 1945 may be added. Capsules may bear capsules. As soft gelatin shells contain appreciable amounts
surface markings. of water, migration of capsule contents, particularly of water-
soluble ingredients, may occur.
The contents of capsules may be of solid, liquid or paste-like
consistency. They consist of the medicament(s) with or without Modified-release Capsules. Modified-release (Sustained-
excipients such as vehicles, solvents, diluents, lubricants, release) Capsules are hard or soft capsules in which the
fillers, wetting agents and disintegrating agents. The contents contents or the shell, or both, contain auxiliary substances or
do not cause deterioration of the shell, but the capsules are are prepared by a special process designed to modify the rate
attacked by the digestive fluids thereby releasing the contents. at which the active ingredients are released.
The contents of capsules other than Modified-release Enteric Capsules (Gastro-resistant Capsules). Enteric
(Sustained-release) Capsules do not contain any added Capsules are hard or soft capsules prepared in such a manner
colouring agent. that the shell resists the action of the gastric fluid but is
attacked by the intestinal fluid to release the contents.
Hard Gelatin Capsules. Hard gelatin capsules contain the
medicament(s) in the solid form. Where two mutually During manufacture, packaging, storage and distribution of
incompatible drugs are present in the mixture, one of the drugs capsules, suitable means shall be taken to ensure their microbial
can be put as a tablet or pellet or in small capsule and then quality; acceptance criteria for microbial quality are given in
enclosed with the other drug in a large capsule. Chapter 5.9.
Production Tests
Hard gelatin capsules are made by a process that involves Content of active ingredients. Determine the amount of active
dipping shaped pins into gelatin solutions, after which the ingredient(s) by the method described in the Assay and
gelatin films are dried, trimmed, and removed from the pins, calculate the amount of active ingredient(s) in each capsule.
and the body and cap pieces are joined. The result lies within the range for the content of active
15
CAPSULES IP 2007
ingredient(s) stated in the monograph. This range is based on NOTE — The test is not applicable for capsules containing
the requirement that 20 capsules, or such other number as multivitamins and trace elements.
may be indicated in the monograph, are used in the Assay. Determine the content of active ingredient in each of 10
Where 20 capsules cannot be obtained, a smaller number, capsules taken at random using the method given in the
which must not be less than 5, may be used, but to allow for monograph or by any other suitable analytical method of
sampling errors the tolerances are widened in accordance with equivalent accuracy and precision. The capsules comply with
Table 1. The requirements of Table 1 apply when the stated the test if not more than one of the individual values thus
limits are between 90 and 110 per cent. For limits other than 90 obtained is outside the limits 85 to 115 per cent of the average
to 110 per cent, proportionately smaller or larger allowances value and none is outside the limits 75 to 125 per cent. If two
should be made or three individual values are outside the limits 85 to 115 per
Table 1 cent of the average value repeat the determination using
another 20 capsules. The capsules comply with the test if in
Weigh of Active Subtract from Add to the upper
the total sample of 30 capsules not more than three individual
ingredients in each the lower limit limit for samples
values are outside the limits 85 to 115 per cent and none is
Capsules for samples of of
outside the limits 75 to 125 per cent of the average value.
15 10 5 15 10 5
Disintegration. The disintegration test is not applicable to
0.12 g or less 0.2 0.7 1.5 0.3 0.8 1.8 Modified-release Capsules. For those Hard Capsules and Soft
More than 0.12 g Capsules for which the dissolution test (2.5.2) is included in
and less than 0.3 g 0.2 0.5 1.2 0.3 0.6 1.5 the individual monograph, the test for Disintegration is not
required.
0.3g or more 0.1 0.2 0.8 0.2 0.4 1.0
Hard Capsules. Comply with the disintegration test (2.5.1).
Uniformity of weight. This test is not applicable to capsules Unless otherwise directed in the individual monograph use
that are required to comply with the test for Uniformity of water as the medium. If the capsules float on the surface of
content for all active ingredients. the medium, a disc may be added. If the capsules adhere to the
Weigh an intact capsule. Open the capsule without losing discs, attach a removable piece of stainless steel woven gauze
any part of the shell and remove the contents as completely with mesh aperture of 2.00 mm to the upper plate of the basket
as possible. To remove the contents of a soft capsule the shell rack assembly and carry out the test omitting the discs. Operate
may be washed with ether or other suitable solvent and the the apparatus for 30 minutes unless otherwise directed.
shell allowed to stand until the odour of the solvent is no Soft Capsules. Comply with the disintegration test (2.5.1).
longer detectable. Weigh the shell. The weight of the contents Unless otherwise directed in the individual monograph use
is the difference between the weighings. Repeat the procedure water as the medium and add a disc to each tube. Operate the
with a further 19 capsules. Determine the average weight. Not apparatus for 60 minutes unless otherwise directed.
more than two of the individual weights deviate from the
Enteric Capsules. Use the apparatus described under
average weight by more than the percentage deviation shown
disintegration test (2.5.1), using one capsule in each tube.
in Table 2 and none deviates by more than twice that
Operate the apparatus for 2 hours without the discs in 0.1 M
percentage.
hydrochloric acid. No capsule shows signs of disintegration
Table 2 or of rupture permitting the escape of the contents. Replace
Average weight of capsule Percentage deviation the medium in the vessel with mixed phosphate buffer pH 6.8,
contents add a disc to each tube and operate the apparatus for a further
60 minutes. Remove the apparatus from the medium and
Less than 300 mg 10 examine the capsules. They pass the test if no residue remains
300mg or more 7.5 on the screen or on the underside of the discs, or, if a residue
remains, it consists of fragments of shell or of a soft mass with
Uniformity of content. This test is applicable to capsules that no palpable, unmoistened core.
contain less than 10 mg or less than 10 per cent w/w of active
ingredient. For capsules containing more than one active Storage. Store at a temperature not exceeding 30º.
ingredient carry out the test for each active ingredient that Labelling. The label states the name of any added antimicrobial
corresponds to the afore-mentioned conditions. preservative.
The test should be carried out only after the content of active
ingredient(s) in a pooled sample of the capsules has been
shown to be within accepted limits of the stated content.
16
IP 2007 EAR DROPS
Creams Sterility. When the cream is labelled as sterile, it complies

with the test for sterility (2.2.11).
Creams are homogeneous, semi-solid or viscous preparations
that possess a relatively fluid consistency and are intended Storage. Store at temperatures below 25º unless otherwise
for external application to the skin or certain mucous directed. Do not freeze.
membranes for protective, therapeutic or prophylactic Labelling. The label states (1) that the cream is sterile, where
purposes especially where an occlusive effect is not necessary. necessary; (2) the name and concentration of any added
They are semisolids usually consisting of solutions or antimicrobial preservative; (3) the storage conditions.
dispersions of one or more medicaments in suitable bases*. * The term basis as a synonym for base in some of the monographs
They are formulated using hydrophilic or hydrophobic bases means a carrier, composed of one or more excipients, for the active
to provide preparations that are essentially miscible with the pharmaceutical ingredient(s) in semi-solid and solid preparations.
skin secretion.
In recent times the term cream has been restricted to products
consisting of oil-in-water emulsions or aqueous Ear Drops
microcrystalline dispersions of long-chain fatty acids or
alcohols that are water-washable and more cosmetically and Otic Drops; Otic Solutions
aesthetically acceptable. Creams can be used for administering
Ear Drops are aqueous or oily solutions or suspensions of
drugs via the vaginal route.
one or more medicaments intended for instillation into the
The base should not produce irritation or sensitisation of the outer ear. They may contain suitable auxiliary substances such
skin, nor should it retard wound healing; it should be smooth, as buffers, stabilising agents, dispersing agents, solubilising
inert, odourless or almost odourless, physically and chemically agents and agents to adjust the tonicity or viscosity of the
stable and compatible with the skin and with incorporated preparation. However, if buffering agents are used in
medicaments. preparations intended for use in surgical procedures, care
Creams may contain suitable antimicrobial preservatives unless should be taken to ensure that the nature and concentration
the active ingredients or the bases themselves have sufficient of the selected agents are suitable. Where the active
bactericidal or fungicidal activity. They may contain other ingredients are susceptible to oxidative degradation, a suitable
suitable auxiliary substances such as antioxidants, stabilisers, antioxidant may be added but care should be taken to ensure
thickeners and emulsifiers. compatibility between the antioxidant and the other ingredients
of the preparations. Any additive in the preparation should
If a cream is specifically intended for use on large open wounds not adversely affect the intended medicinal action nor, at the
or on severely injured skin it should be sterile. concentrations used, cause undue local irritation. Certain Ear
Creams should not normally be diluted; should dilution be Drops may be supplied in dry, sterile form to be constituted in
necessary care should be taken to prevent instability and, in an appropriate sterile liquid immediately before use.
particular, microbial contamination. Aqueous preparations supplied in multiple application
containers contain suitable antimicrobial preservatives at
Production appropriate concentrations except when the product itself has
Creams should be packed in well-closed containers fitted with adequate antimicrobial properties. The antimicrobial
closures that minimise contamination with micro-organisms. preservatives should be compatible with the other ingredients
When practicable, creams should be packed in collapsible of the preparation and should be effective throughout the
tubes of suitable metal or plastic. period of use of the Ear Drops. Containers for multiple
application preparations should permit the withdrawal of
During manufacture, packaging, storage and distribution of
successive doses of the preparation. Such containers should
creams, suitable means shall be taken to ensure their microbial
normally hold not more than 10 ml.
quality; acceptance criteria for microbial quality are given in
Chapter 5.9. During development of a formulation of ear drops containing
an antimicrobial preservative, the need for and the efficacy of
Tests the chosen preservative shall be demonstrated by the test for
Creams comply with the requirements of tests stated under efficacy of antimicrobial preservation (2.2.2).
the individual monographs and with the following During manufacture, packaging, storage and distribution of
requirements. ear drops, suitable means shall be taken to ensure their
Uniformity of weight. Comply with the test for contents of microbial quality; acceptance criteria for microbial quality are
packaged dosage forms (2.5.6). given in Chapter 5.9.
17
EYE DROPS IP 2007
Ear Drops intended for use in surgical procedures or for Labelling. The label states (1) the names and concentrations
application to injured ear, are sterile. Such preparations should in percentages, or weight or volume per ml, of the active
not contain antimicrobial preservatives and should be packed ingredient(s); (2) the names and concentrations of any added
in single dose containers. antioxidant, stabilising agent or antimicrobial preservative;
(3) that, for multiple application containers, the contents
Production should not be used for more than 1 month after opening the
container; (4) that, for multiple application containers, care
Sterile Ear Drops are prepared using methods designed to
should be taken to avoid contamination of the contents during
ensure their sterility and to avoid the introduction of
use; (5) that the preparation is NOT FOR INJECTION; (6) that,
contaminants and growth of micro-organisms. Methods of
where applicable, the preparation is sterile; (7) the storage
sterilisation that may be used in the manufacture of Ear Drops
conditions.
are described in Chapter 5.3.
Description. Ear Drops that are solutions are practically clear
and practically free from particles when examined under Eye Drops
suitable conditions of visibility. Ear Drops that are suspensions
Ophthalmic Drops
may show a sediment that readily disperses when shaken.
The suspension remains sufficiently dispersed to enable the Eye Drops are sterile, aqueous or oily solutions or suspensions
correct dose to be removed from the container. of one or more medicaments intended for instillation into the
conjunctival sac. They may contain suitable auxiliary
Tests substances such as buffers, stabilising agents, solubilising
agents and agents to adjust the tonicity or viscosity of the
Uniformity of volume. Comply with the test for contents of
preparation. However, if buffering agents are used in
packaged dosage forms (2.5.6).
preparations intended for use in surgical procedures care
Particle size. This test is applicable only to Ear Drops that are should be taken to ensure that the nature and concentration
suspensions. Introduce a suitable volume of the Ear Drops of the selected agents are suitable. Where the active ingredient
into a counting cell or onto a microscope slide, as appropriate. is susceptible to oxidative degradation, a suitable antioxidant
Scan under a microscope an area corresponding to 10 µg of may be added but care should be taken to ensure compatibility
the solid phase. Scan at least 50 representative fields. Not between the antioxidant and the other ingredients of the
more than 20 particles have a maximum dimension greater than preparation. Any additive in the preparation should not
25 µm, not more than 10 particles have a maximum dimension adversely affect the intended medicinal action nor, at the
greater than 50 µm and none has a maximum dimension greater concentrations used, cause undue local irritation. Certain Eye
than 100 µm. Drops may be supplied in dry, sterile form to be constituted in
Sterility. Where the label indicates that the Ear Drops are an appropriate sterile liquid immediately before use.
sterile, it complies with the test for sterility (2.2.11). Droppers Aqueous preparations supplied in multiple application
supplied separately also comply with these tests. Remove the containers contain suitable antimicrobial preservatives at
dropper out of the package using aseptic precautions and appropriate concentrations except when the product itself has
transfer it to a tube containing suitable culture medium so that adequate antimicrobial properties. The antimicrobial
it is completely immersed. Incubate and carry out the tests for preservatives should be compatible with the other ingredients
sterility on the medium. of the preparation and should be effective throughout the
Storage. Ear Drops should be packed in well-closed containers. period of use of the Eye Drops.
If the preparation is sterile, store in sterile, tightly-closed, If the preparation does not contain an antimicrobial
tamper-evident containers. Containers should be made from preservative it should be packed in single application
materials that do not cause deterioration of the preparation as containers. Eye Drops intended for use in surgical procedures
a result of diffusion into or across the material of the container should not contain antimicrobial preservatives and should be
or by yielding foreign substances to the preparation. packed in single application containers.
The container and package of a single application preparation Eye Drops are prepared using methods designed to ensure
should be such as to maintain sterility of the contents and the their sterility and to avoid the introduction of contaminants
applicator up to the time of use. Containers for multiple and growth of micro-organisms. Methods of sterilisation that
application preparations should be fitted with an integral may be used in the manufacture of Eye Drops are described in
dropper or with a screw cap made of suitable material chapter 5.3.
incorporating a dropper and plastic or rubber teat. Containers. Eye Drops should be packed in tamper-evident
Alternatively, such a cap assembly may be packed separately. containers. Containers should be made from materials that do
18
IP 2007 GELS
not cause deterioration of the preparation as a result of Eye Ointments

diffusion into or across the material of the container or by
yielding foreign substances to the preparation. Ophthalmic Ointments
The container and package of a single dose preparation should Eye Ointments are sterile, semi-solid preparations of
be such as to maintain sterility of the contents and the homogenous appearance intended for application to the eye.
applicator up to the time of use. Containers for multiple They may contain one or more medicaments dissolved or
application preparations should be fitted with an integral dispersed in a suitable basis. Bases, which are usually non-
dropper or with a sterile screw cap of suitable materials aqueous, may contain suitable auxiliary substances such as
incorporating a dropper and plastic or rubber teat. stabilising agents, antimicrobial preservatives and
Alternatively, such a cap assembly may be packed separately antioxidants. The base selected must be non-irritant to the
after it is sterilised. Containers of multiple application conjunctiva, allow the drug to diffuse throughout the
preparations should permit the withdrawal of successive doses secretions of the eye and retain the activity of the medicaments
of the preparation. Such containers should normally hold not for a reasonable period of time under the stated conditions of
more than 10 ml. storage.
Description. Eye Drops that are solutions are practically clear Eye Ointments are prepared using methods designed to ensure
and practically free from particles when examined under their sterility and to avoid the introduction of contaminants
suitable conditions of visibility. Eye Drops that are and growth of micro-organisms. Methods of sterilisation that
suspensions may show a sediment that readily disperses when may be used in the manufacture of Eye Ointments are described
shaken. The suspension remains sufficiently dispersed to in Chapter 5.3.
enable the correct dose to be removed from the container. Containers. Eye Ointments should be packed in small,
sterilised collapsible tubes of metal or of suitable plastic fitted
Tests
or provided with a nozzle of suitable shape to facilitate the
Uniformity of volume. Comply with the test for contents of application of the product without contamination and with a
packaged dosage forms (2.5.6). cap. The content of such containers is not more that 5 g of the
preparation. Eye Ointments may also be packed in single
Particle size. This test is applicable only to Eye Drops that
application containers of such a shape as to facilitate
are suspensions. Introduce a suitable volume of the Eye Drops
administration without contamination; such containers may
into a counting cell or onto a microscope slide, as appropriate.
be individually wrapped. Other requirements concerning
Scan under a microscope an area corresponding to 10 µg of
containers are given in Chapter 6.2.
the solid phase. Scan at least 50 representative fields. Not
more than 20 particles have a maximum dimension greater than Tests
25 µm, not more than 10 particles have a maximum dimension
greater than 50 µm and none has a maximum dimension greater Uniformity of weight. Comply with the test for contents of
than 100 µm. packaged dosage forms (2.5.6).
Sterility. Comply with the test for sterility (2.2.11). Droppers Particle size. Gently spread a small quantity of the Eye
supplied separately also comply with these tests. Remove the Ointment as a thin layer on a microscope slide. Scan under a
dropper out of the package using aseptic precautions and microscope an area corresponding to 10 µg of the solid phase.
transfer it to a tube containing suitable culture medium so that Scan at least 50 representative fields. Not more that 20 particles
it is completely immersed. Incubate and carry out the test. have a maximum dimension greater than 25 µm, not more than
10 particles have a maximum dimension greater than 50 µm
Storage. Store in sterile containers sealed so as to protect
and none has a maximum dimension greater than 100 µm.
from micro-organisms.
Sterility (2.2.11). Comply with the test for sterility.
Labelling. The label states (1) the names and concentrations
in percentages, or weight or volume per ml, of the active Storage. Store at temperatures below 30º unless otherwise
ingredients; (2) the names and concentrations of any added directed. Do not freeze.
antimicrobial preservative; (3) that, for multiple application
containers, the contents should not be used for more than 1
month after opening the container; (4) that, for multiple
application containers, care should be taken to avoid Gels
contamination of the contents during use; (5) that the Gels are homogeneous, semi-solid preparations usually
preparation is NOT FOR INJECTION; (6) the conditions under consisting of solutions or dispersions of one or more
which the preparation should be stored. medicaments in suitable hydrophilic or hydrophobic bases.
19
INHALATION PREPARATIONS IP 2007
They are normally prepared with the aid of suitable gelling preservative selected, shall be determined as described in
agents. They are intended to be applied to the skin or certain chapter 2.2.2 (Efficacy of antimicrobial preservation).
mucous membranes for protective, prophylactic or therapeutic
The size of aerosol particles shall be controlled so that a
purposes. Gels may contain suitable added substances such
significant fraction is deposited in the lung.
as antioxidants, stabilisers and antimicrobial preservatives.
The most commonly used method of preparation involves
filling under pressure and sometimes by filling after
gels, suitable means shall be taken to ensure their microbial
refrigeration to temperatures below 0º. In filling under pressure,
the requisite volume of the concentrate of the active
Chapter 5.9.
ingredient(s) is filled in the container and either the propellant
Gels specifically intended for use on large open wounds or on is forced under pressure through the valve orifice after the
severely injured skin should be sterile. valve is sealed, or the propellant is allowed to flow under the
Containers. Gels should be packed in suitable well-closed or, valve cap and the valve assembly is sealed. In either case, the
if the preparation contains water or other volatile ingredients, air in the container must be evacuated by means of vacuum or
suitable tightly-closed containers. The containers should be displacement with a small amount of the propellant.
fitted with closures that minimise contamination with micro- During production, strict control should be exercised by
organisms. To the extent possible, collapsible tubes of suitable process controls that include propellant and medicament fill
metal or plastic should be used. weights, pressure test and leak test of the finished product.
Storage. Store at temperatures below 30º unless otherwise For preparations adversely affected by water present in
directed. Do not freeze. quantities beyond certain limits, care should be taken to protect
Labelling. The label states (1) that the gel is sterile, where the products from moisture.
necessary; (2) the storage conditions. Storage. Avoid storage under extremes of temperature and in
Tests an environment with undue fluctuations in temperature.
Uniformity of weight. Comply with the test for contents of Labelling. The label states (1) the name(s) of the active
packaged dosage forms (2.5.6). ingredient(s); (2) the total amount of the active ingredient(s)
in the container except in the case of metered-dose preparation
Sterility. Gels labelled as sterile comply with the test for sterility
for inhalation); (3) that the container should be shaken before
(2.2.11).
use; (4) the other instructions for use; (5) the date after which
the preparation is not intended to be used; (6) the conditions
under which it should be stored; (7) a warning that the
Inhalation Preparations container is under pressure and that it must not be punctured,
Inhalation Preparations are liquid or solid dosage forms broken or incinerated even when apparently empty; (8) the
intended for administration as vapours or aerosols to the lung statement. “Warning. Keep away from children”
in order to obtain a local or systemic effect. They contain In the case of metered-dose aerosols and pressurized metered
solutions or dispersions of one or more active ingredients dose inhalers, the label states in addition (1) the total number
which may be dissolved or dispersed in a suitable vehicle. of deliveries available from the container; (2) the amount of
Inhalation Preparations contain propellants, diluents, active ingredient(s) released each time the valve is actuated.
antimicrobial agents, solubilising and stabilising agents etc.
In the case of dry powder inhalers the label on the container
depending on the type of preparation. They are available in
states (1) the date after which the dry powder inhaler is not
single-dose or multidose containers.
intended to be used; (2) the conditions under which the powder
Inhalation Preparations intended to be administered as for Inhalation should be stored. Where the powder for
aerosols (dispersions of solid or liquid particles of active Inhalation is supplied in a capsule, the label also states (3)
ingredient(s) in a gas) are administered by pressurized the quantity of the active ingredient contained in each capsule;
metered-dose inhalers or by powder inhalers. (4) that the capsules are intended for use in an inhaler and are
not to be swallowed.
Production
Information on use of the preparation provided in the pack
Inhalation preparations should be manufactured in conditions shall include (1) the direction for correct use of the aerosol; (2)
designed to minimise microbial and particulate contamination. a warning that the container may explode if punctured, exposed
During the development of a preparation that contains an to excessive heat or direct sunlight; (3) the directions for the
antimicrobial preservative, the effectiveness of the disposal of the used or partly-used container.
20
IP 2007 INHALATION PREPARATIONS
Pressurised metered-dose preparations are solutions, Metered valves may need priming before use if the aerosol
suspensions or emulsions supplied in containers equipped packages have not been stored properly or have not been
with a metering valve and which are held under pressure with used for long periods of time.
suitable propellants or mixtures of liquefied propellants. Actuators. The actuator or adaptor which is fitted to the aerosol
Pressurised Metered Dose Inhalers are dosage forms valve stem is a device which on depression or any other
containing therapeutically active ingredients that are packaged required movement opens the valve and directs the spray to
under pressure in a sealed container and are released as a fine the desired area. The design of the actuator which incorporates
mist of spray upon activation of a suitable valve system. an orifice of varying size and shape and expansion chamber is
very important in influencing the physical characteristics of
The basic components of an aerosol system are the container,
the spray or foam, particularly in the case of inhalation
the propellant, the concentrate containing the active
aerosols, where the active ingredient(s) must be delivered in
ingredient(s), the valve and the actuator.
the proper particle size range. A proportion of the active
Pressurised metered dose preparations are of two types, the ingredient(s) is usually deposited on the inner surface of the
two-phase system consisting of gas and liquid or the three- actuator; the amount available is therefore less than the amount
phase system consisting of gas, liquid and solid or liquid. The released by actuation of the valve.
two-phase preparation comprises a solution of active
Containers. Aerosol containers are made of metal (stainless
ingredient(s) in liquefied propellant and the vaporised
steel, aluminum or tin-plated steel), glass or plastic or a
propellant. The solvent is usually the propellant or a mixture
combination of these materials. The containers must be so
of the propellant and co-solvents such as ethanol, propylene
designed that they provide the maximum in pressure safety
glycol and polyethylene glycols. The three-phase preparation
and impact resistance.
consists of a suspension or emulsion of the active ingredient(s)
and the vaporised propellants. In the suspension the Tests
ingredient(s) may be dispersed in the propellant system with
the aid of suitable pharmaceutical aids such as wetting agents, Pressurised Metered-dose Preparations
solubilising agents, emulsifying agents, suspending agents
and lubricating agents to prevent clogging of valves. Content of active ingredient delivered per actuation.
Active ingredients. For satisfactory bioavailability the active
Apparatus
ingredient(s) should have the majority of particles under 10
µm in size in the case of inhalation aerosols and not more than A small sample vessel suitable for shaking. The size of the
100 µm for other types of aerosols. vessel is such that when the aerosol is discharged into the
Propellants. For pressurised metered dose inhalations specified volume of solvent under the conditions described
propellants perform the essential function of expelling the in the Method below, the discharge takes place not less than
material from the container by supplying the necessary 25 mm below the surface of the solvent. A stainless steel base
pressure within the aerosol system. They are liquefied or plate with 3 legs and a central circular indentation with a hole
compounded gases having vapour pressures exceeding about 1.5 mm in diameter is placed in the sample vessel. The
atmospheric pressure. The commonly used propellants in arrangement should prevent particle entrapment and side-of-
aerosol systems are hydrocarbons, especially the fluorochloro- stem leakage during the delivery of the sample.
derivatives of methane and ethane, the butanes and pentanes Procedure
and compressed gases such as nitrogen and carbon dioxide.
Mixtures of propellants are often employed to obtain the Remove the pressurised container from the actuator and remove
necessary delivery and spray characteristics of the aerosol. all labels and markings which may be present on the container
with a suitable solvent. Dry the container, replace in its actuator,
Valves. The valve regulates the flow of the active ingredient(s)
shake for about 30 seconds and holding it in an inverted
and propellant from the container and determines the spray
position actuate the valve by discharging about 5 sprays to
characteristics of the aerosol. It must be manufactured from
waste. Remove the pressurised container from its actuator,
materials which are inert to the contents of the aerosol. The
clean the valve stem (internally and externally) and valve
commonly used materials are rubber, plastic, aluminium and
ferrule by washing with a suitable solvent. Dry the complete
stainless steel.
valve assembly using an air-supply line fitted with an
Products for oral or nasal inhalation require metered-dose appropriate narrow jet to ensure that all solvent is removed
valves which ensure delivery of a uniform quantity of spray from the inside of the valve stem. Wash the actuator after the
and an accurate dose of the active ingredient(s), both within initial discharge of 5 sprays to waste, with a suitable solvent
specified tolerances, with each activation of the valve. and allow it to dry.
21
For test solution add to the sample vessel a volume of solvent position, an equivalent test is applied using methods that
or solvent mixture specified in the monograph so that the final ensure the complete collection of the delivered dose.
concentration of the active ingredient in the test solutin For all the cases, prepare the inhaler as directed in the
corresponds to the reference solution. Shake the pressurised instructions to the patient and connect to a dose collection
container for about 30 seconds and place it inverted in the apparatus, which must be capable of quantitatively capturing
vessel. Discharge 10 deliveries below the surface of the solvent the delivered dose (see Fig.1).
actuating the valve at intervals of not less than 5 seconds,
maintaining the pressurised container in the vertical plane The apparatus consists of a filter-support base with an open-
and discharging the aerosol through the hole in the centre of mesh filter-support, such as a stainless steel screen, a sample
the base plate. With some preparations it may be necessary to collection tube that is clamped or screwed to the filter-support
shake the pressurised container between each actuation of base, and a mouthpiece adapter to ensure an airtight seal
the valve; in such cases shaking should be carried out without between the sample collection tube and the mouthpiece. Use
removing the pressurised container from its inverted position a mouthpiece adapter which ensures that the front face of the
in the vessel. Remove the pressurised container, wash it with inhaler mouthpiece fits with the front face or the 2.5
the specified solvent and dilute the combined solution and mm indented shoulder of the sample collection tube, as
washings to the volume specified in the monograph. Determine appropriate. The vacuum connector is connected to a system
the amount of active ingredient by the method described under comprising a vacuum source and a flow regulator. The source
Assay in the individual monograph This amount of active should be adjusted to draw air through the complete
ingredient is referred as metered dose assay (A) for metered assembly, including the filter and the inhaler to be tested, at
dose inhalers. 28.3 litres per minutes (± 5 per cent). Air should be drawn
continuously through the apparatus to avoid loss of the
Fit the washed and dried actuator to the pressurised container
active substance into the atmosphere. The filter-support base
and actuate the valve 10 times at intervals of not less than 5
is designed to accommodate 25 mm diameter filter disks.
seconds. Remove the actuator carefully from the pressurised
container and wash it with small quantities of the specified The filter disk and other materials used in the construction of
solvent or solvent mixture. Dilute the combined washings the apparatus must be compatible with the active substance
suitably and on the resulting solution determine the amount and solvents that are used to extract the active substance
of active ingredient as per the method given in the individual from the filter.
monograph under the test for ‘Content of active ingredient One end of the collection tube is designed to hold the filter
delivered per actuation’ and calculate the amount of active disk tightly against the filter-support base. When assembled,
ingredient per actuation of the valve. This amount of active the joints between the components of the apparatus are airtight
ingredient is referred to as actuator retention (B) for metered so that when a vacuum is applied to the base of the filter, all of
dose inhalers. the air drawn through the collection tube passes through the
Calculate the content of active ingredient delivered per inhaler.
actuation from the expression A – B.
Procedure
Uniformity of delivered dose Unless otherwise prescribed in the instructions to the patient,
The delivered dose is the dose delivered from the inhaler to shake the inhaler for 5 seconds and discharge one delivery to
the patient. For some preparations, the dose has been waste. Attach the inverted inhaler to the apparatus, depressing
established as a metered dose. The metered dose is determined the valve for a sufficient time to ensure complete discharge.
by adding the amount deposited on the inhaler device to the Repeat the procedure until the number of deliveries that
delivered dose. It may also be determined directly. constitute the minimum recommended dose have been
sampled. Quantitatively collect the contents of the apparatus
The test is applicable to inhalation preparations containing and determine the amount of active substance.
the drug formulation (e.g., solution, suspension, or powder)
either in reservoirs or in premetered dosage units, and for Repeat the procedure for a further 2 doses.
drug formulations packaged in reservoirs or in premetered Discharge the device to waste, waiting not less than 5 seconds
dosage units where these containers are labeled for use with between actuations until (n/2) +1 deliveries remain, where n is
a named inhalation device. the number of deliveries stated on the label. Collect 4 doses
using the procedure described above.
Apparatus
Discharge the device to waste, waiting not less than 5 seconds
Most of the containers usually operate in a valve-down between actuations until 3 doses remain. Collect these 3 doses
position. For those containers that operate in a valve-up using the procedure described above.
22
Dimensions in millimeters
Fig. 1: Dose collection apparatus for pressurized metered-dose inhalers
For preparations containing more than one active substance, Acceptance criteria
carry out the test for uniformity of delivered dose for each Unless otherwise justified and authorised, the preparation
active substance. complies with the test if 9 out of 10 results lie between 75 per
23
cent and 125 per cent of the average value and all lie between reduce the internal pressure, removing the valve and pouring.
65 per cent and 135 per cent. If 2 or 3 values lie outside the Remove any residual contents by rinsing with suitable
limits of 75 per cent to 125 per cent, repeat the test for 2 more solvents, then rinse with a few portions of methanol. Retain
inhalers. Not more than 3 of the 30 values lie outside the limits as a unit the container, the valve, and all associated parts, and
of 75 per cent to 125 per cent and no value lies outside the heat them at 100º for 5 minutes. Cool, weigh and record the
limits of 65 per cent to 135 per cent. weight as W3, and determine the net fill weight (W1-W3) for
each container tested.
Particle size
The requirements are met if the average leakage rate of the 12
NOTE — Carry out the test in a laminar flow cabinet. Filter containers is not more than 3.5 per cent of the net fill weight
all solvents through an appropriately sized filter before use. per year and none of the containers leaks more than 5.0 per
Assemble a suitable membrane filtration apparatus. Use a filter cent of the net fill weight per year. If 1 container leaks more
holder fitted with an input chamber designed to prevent any than 5.0 per cent per year, and if none of the containers leaks
loss of material when the actuator mouthpiece of the aerosol more than 7.0 per cent per year, determine the leakage rate of
is inserted and the valve actuated. Before assembly wash all an additional 24 containers as directed herein. Not more than
parts of the membrane filter holder with water and methanol 2 of the 36 containers leak more than 7.0 per cent of the net fill
and dry in a stream of nitrogen or allow to dry in a laminar flow weight per year.
cabinet. Use a membrane filter with a nominal pore size not Where the net fill weight is less than 15 g the requirements are
greater than 5 µm and with the filtering surface free from foreign met if the average leakage rate of the 12 containers is not more
particles when examined microscopically using a magnification than 525 mg per year and none of the container leaks more
of not less than × 40. than 750 mg per year. If 1 container leaks more than 750 mg per
Discharge 50 deliveries from the pressurised container into year but not more than 1.1 g per year, determine the leakage
the orifice of the input chamber, actuating the valve at intervals rate of an additional 24 containers as directed herein. Not
of not less than 5 seconds and washing down the particles more than 2 of the 36 containers leak more than 750 mg per
deposited in the input chamber with successive 10-ml year and none of the 36 containers leaks more than 1.1 g per
quantities of light petroleum (40º to 60º), ethanol (95 per year.
cent) and water after 20, 40 and 50 actuations of the valve.
Remove the pressurised container and dry the membrane filter. Deposition of the emitted dose
Examine its entire filtering surface microscopically using a The deposition of the emitted dose is a measure of the drug
magnification of not less than x40. Record the number and deposition during inhalation. This test is used to determine
size of all individual particles (not agglomerates) more than 10 the fine particle characteristics of the aerosol clouds generated
µm in length measured along the longest axis. The number of by preparations for inhalation and may be expected to correlate
particles longer than 20 µm does not exceed 50 and no particle with the drug dose or that fraction of the drug dose that
exceeds 100 µm in length. penetrates the lung during inhalation. Individual monographs
Number of deliveries per container. Take the pressurised may also define the emitted fractions of the delivered dose in
container used in the test for Particle size and discharge the more than one particle size range.
remaining contents to waste, actuating the valve at intervals Stage Mensuration. Manufacturers of cascade impaction
of not less than 5 seconds. Record the number of deliveries devices provide a definitive calibration for the separation
discharged. The total number of deliveries so discharged in characteristics of each impaction stage in terms of the
the test for Particle size is not less than the number stated on relationship between the stage collection efficiency and the
the label. aerodynamic diameter of particles and droplets passing
Leak test. Select 12 pressurised containers at random, and through it as an aerosol. Calibration is a property of the jet
record the date and time to the nearest half-hour. Weigh each dimensions, the spatial arrangement of the jet and its collection
container to the nearest mg, and record the weight, in mg, of surface, and the airflow rate passing through it. Because jets
each as W1. Allow the container to stand in an upright position can corrode and wear over time, the critical dimensions of
at room temperature for not less than 3 days, and again weigh each stage, which define that impaction stage’s calibration,
each container, recording the weight, in mg, of each as W2 and must be measured on a regular basis. This process, known as
recording the date and time to the nearest half-hour. Determine stage mensuration, replaces the need for repetitive calibration
the time, T, in hours, during which the containers were under (using standard aerosols) and ensures that only devices that
test. Calculate the leakage rate, in mg per year, of each container conform to specifications are used for testing inhaler output.
from the expression 365 x 24/T x (W1 - W2). The process involves the measurement and adjustment of the
critical dimensions of the instrument.
Empty the contents of each container tested by chilling to
Re-entrainment (for apparatus B). To ensure efficient particle
24
capture, coat each plate with glycerol, silicone oil or similar depth of about 10 mm, lines up along the horizontal axis of the
high viscosity liquid, typically deposited from a volatile throat and the open end of the actuator, which accepts
solvent. Plate coating must be part of method validation the pressurised container, is uppermost and in the same vertical
and may be omitted where justified and authorised. plane as the rest of the apparatus.
Mass balance. The total mass of the active substance is not Introduce 7 ml and 30 ml of a suitable solvent into the upper
less than 75 per cent and not more than 125 per cent of the and lower impingement chambers, respectively.
average delivered dose determined during testing for
Connect all the component parts. Ensure that the assembly is
uniformity of delivered dose. This is not a test of the inhaler
vertical and adequately supported and that the lower jet-spacer
but it serves to ensure that the results are valid.
peg of the lower jet assembly just touches the bottom of the
Unless otherwise specified, one of the following apparatus lower impingement chamber. Connect a suitable pump to the
and test procedures is used. outlet of the apparatus. Adjust the air flow through the
Apparatus A. Glass impinger apparatus, as measured at the inlet to the throat, to 60 ± 5
litres per minute.
The apparatus is shown in Fig. 2 and the dimensions are given
in Table 1. Prime the metering valve by shaking for 5 seconds and
discharging once to waste; after not less than 5 seconds, shake
Procedure and discharge again to waste. Repeat for further 3 times.
Place the actuator adapter in position at the end of the throat Shake for about 5 seconds, switch on the pump to the
so that the mouthpiece end of the actuator, when inserted to a apparatus and locate the mouthpiece end of the actuator in
Dimensions in millimeters (tolerances ± 1 mm, unless otherwise specified)

Fig. 2: Apparatus A. Glass impinger
25
Table 1
Code Item Description Dimensions
A Mouthpiece adaptor Moulded rubber adapter for actuator mouthpiece.
B. Throat Modified round-bottomed flask: 50 ml
ground-glass inlet socket 29/32
ground-glass outlet cone 24/29
C. Neck Modified glass adapter:
Lower outlet section of precision-bore glass tubing:
bore diameter 14
Selected bore light-wall glass tubing:
external diameter 17
D. Upper impingement chamber Modified round-bottomed flask 100 ml
E. Coupling tube Medium-wall glass tubing:
ground-glass cone 14/23
Bent section and upper vertical section:
external diameter 13
Lower vertical section:
external diameter 8
F. Screw thread, side-arm adaptor Plastic screw cap 28/13
Silicone rubber ring 28/11
PTFE washer 28/11
Glass screw thread:
thread size 28
Side-arm outlet to vacuum pump:
minimum bore diameter 5
G. Lower jet assembly Modified polypropylene filter holder See Figure1
connected to lower vertical section of
coupling tube by PTFE tubing
Acetal circular disc with the centres of four jets 10
arranged on a projected circle of diameter 5.3 mm
with an integral jet spacer peg:
peg diameter 2
peg protrusion 2
H. Lower impingement chamber Conical flask 250 ml
the adapter, discharge once immediately. Remove sequence. The number of discharges should be minimised
the assembled inhaler from the adapter, shake for not less and typically would not be greater than 10. After the final
than 5 seconds, relocate the mouthpiece end of the actuator discharge wait for not less than 5 seconds and then switch off
in the adapter and discharge again. Repeat the discharge the pump. Dismantle the apparatus.
26
Wash the inner surface of the inlet tube to the lower induction port (see Fig. 4). A suitable mouthpiece adapter is
impingement chamber and its outer surface that projects into used to provide an airtight seal between the inhaler and the
the chamber with a suitable solvent, collecting the washings induction port. The front face of the inhaler mouthpiece must
in the lower impingement chamber. Determine the content of be flush with the front face of the induction port.
active substance in this solution. Calculate the amount of Table 2 - Critical dimensions for Apparatus B
active substance collected in the lower impingement chamber
per discharge and express the results as a percentage of the Description Number Dimension (mm)
dose stated on the label. Stage 0 nozzle diameter 96 2.55 ± 0.025
Apparatus B. Andersen Cascade impactor Stage 1 nozzle diameter 96 1.89 ± 0.025
Stage 2 nozzle diameter 400 0.914 ± 0.0127
The Andersen 1 ACFM non-viable cascade impactor consists
of 8 stages together with a final filter. Material of construction Stage 3 nozzle diameter 400 0.711 ± 0.0127
may be aluminium, stainless steel or other suitable material. Stage 4 nozzle diameter 400 0.533 ± 0.0127
The stages are clamped together and sealed with O-rings. Stage 5 nozzle diameter 400 0.343 ± 0.0127
Critical dimensions applied by the manufacturer of apparatus
Stage 6 nozzle diameter 400 0.254 ± 0.0127
B are provided in Table 2. In use, some occlusion and wear of
holes will occur. In-use mensuration tolerances need to be Stage 7 nozzle diameter 201 0.254 ± 0.0127
justified. In the configuration used for pressurised inhalers In the configuration for powder inhalers, a pre-separator is
(Fig. 3) the entry cone of the impactor is connected to an placed above the top stage to collect large masses of non-
Fig. 3: Apparatus B. Anderson cascade impactor
27
respirable powder. It is connected to the induction port as Procedure

shown in Fig. 5. To accommodate high flow rates through
the impactor, the outlet nipple, used to connect the impactor Assemble the Andersen impactor with a suitable filter in place.
to the vacuum system is enlarged to have an internal diameter Ensure that the system is airtight. In that respect, follow the
of greater than or equal to 8 mm. manufacturer’s instructions. Place a suitable mouthpiece
Note:
1. Material may be aluminium, stainless steel or other suitable material.
2. Machine from 38 mm bar stock.
3. Bore 19 mm hole through bar.
4. Cut tube to exact 45º as shown.
5. The inner bores and tapers should be smooth – surface roughness Ra approx. 0.4 µm.
6. Mill joining cads of stock to provide a liquid tight leak-free seal.
7. Set up a holding fixture for aligning the inner 19 mm bore and for drilling and tapping M4 x 0.7 threads. There must
be virtually no mismatch of the inner bores in the miter joint.
Dimensions in millimeters unless otherwise stated

Fig.4: Induction port
28
adapter in position at the end of the induction port so that the end of the actuator in the adapter and discharge the inverted
mouthpiece end of the actuator, when inserted, lines up along inhaler into the apparatus, depressing the valve for a sufficient
the horizontal axis of the induction port and the inhaler unit is time to ensure complete discharge. Wait for 5 seconds before
positioned in the same orientation as the intended use. Connect removing the assembled inhaler from the adapter. Repeat the
a suitable pump to the outlet of the apparatus and adjust procedure. The number of discharges should be minimised
the air flow through the apparatus, as measured at the inlet to and typically would not be greater than 10. The number of
the induction port, to 28.3 litres per minute (± 5 per cent). discharges is sufficient to ensure an accurate and precise
Switch off the pump. determination of the fine particle dose. After the final
Unless otherwise prescribed in the patient instructions, shake discharge, wait for 5 seconds and then switch off the pump.
the inhaler for 5 seconds and discharge one delivery to waste. Dismantle the apparatus. Carefully remove the filter and extract
Switch on the pump to the apparatus, locate the mouthpiece the active substance into an aliquot of the solvent. Remove
(Dimensions are in millimeters unless otherwise stated)

Fig. 5: Connection of the induction port to the preseparator of the Andersen cascade impactor
29
the induction port and mouthpiece adapter from the apparatus the inhaler is loaded with powders pre-dispensed in capsules
and extract the active substance into an aliquot of the solvent. or other suitable pharmaceutical forms. For inhalers using a
Extract the active substance from the inner walls and the powder reservoir, the dose is created by a metering mechanism
collection plate of each of the stages of the apparatus into within the inhaler.
aliquots of solvent. They are intended either for inhalation for local action in the
Using a suitable method of analysis, determine the quantity lungs or for systemic absorption through the alveoli or for
of active substance contained in each of the aliquots of topical application to the skin or various body orifices.
solvent. Inhalation aerosols are metered dose preparations which
provide controlled amounts of the active ingredient(s).
Calculate the fine particle dose as described below.
Tests
Calculations
From the analysis of the solutions, calculate the mass of active Uniformity of delivered dose
substance deposited on each stage per discharge and the
mass of active substance per discharge deposited in the Procedure
induction port, mouthpiece adapter and when used, the pre-
Prepare the inhaler as directed in the instructions to the patient.
separator.
The dose collection apparatus must be capable of
Starting at the final collection site (filter or MOC), derive a quantitatively capturing the delivered dose. A dose collection
table of cumulative mass versus cut-off diameter of the apparatus similar to that described for the evaluation
respective stage (see Table 3). Calculate by interpolation of pressurised metered-dose inhalers may be used provided
the mass of the active substance less than 5 µm. This is the that the dimensions of the tube and the filter can accommodate
Fine Particle Dose (FPD). the measured flow rate. A suitable tube is defined in Table 4.
Connect the tube to a flow system according to the
If necessary, and where appropriate (e.g., where there is a log-
scheme specified in Fig. 6 and Table 4.
normal distribution), plot the cumulative fraction of active
substance versus cut-off diameter (see Table 4) on log Unless otherwise stated, determine the test flow rate and
probability paper, and use this plot to determine values for duration using the dose collection tube, the associated flow
the Mass Median Aerodynamic Diameter (MMAD) and system, a suitable differential pressure meter and a suitable
Geometric Standard Deviation (GSD) as appropriate. volumetric flowmeter, calibrated for the flow leaving the
Appropriate computational methods may also be used. meter, according to the following procedure.
Powders for Inhalation Prepare the inhaler for use and connect it to the inlet of the
apparatus using a mouthpiece adapter to ensure an airtight
Powders for inhalation are presented as single-dose powders seal. Use a mouthpiece adapter which ensures that the front
or multidose powders. To facilitate their use, active substances face of the inhaler mouthpiece fits with the front face of
may be combined with a suitable carrier. They are generally the sample collection tube. Connect one port of a differential
administered by powder inhalers. For pre-metered inhalers, pressure meter to the pressure reading point, P1, in Figure 6
Table 3 – Calculations for apparatus D when used at a flow rate of 28.3 litres/min
Cut-off diameter Mass of active substance Cumulative mass of Cumulative fraction of
deposited per discharge active substance active substance
(ì m) deposited per discharge (per cent)
d7 = 0.4 mass from stage 8, m8 c7 = m8 f7 = (c7/c) × 100
d6 = 0.7 mass from stage 7, m7 c6 = c7 + m7 f6 = (c6/c) × 100
mass from stage 0, m0 c = c0 + m0 100
30
Table 4 – Specifications of the apparatus shown in Fig. 6

Code Item Description
A Sample collection tube Capable of quantitatively capturing the delivered dose, e.g. dose collection
tube similar to that described in Figure A with dimensions of 34.85 mm ID
x 12 cm length (e.g. product number XX40 047 00, Millipore Corporation,
Bedford, MA 01732 with modified exit tube, ID > 8 mm, fitted with Gelman
product number 61631), or equivalent.
B Filter 47 mm filter, e.g. A/E glass fibre filter (Gelman Sciences, Ann Arbor, MI
48106), or equivalent.
C Connector ID > 8 mm, e.g. short metal coupling, with low-diameter branch to P3
D Vacuum tubing A length of suitable tubing having an ID > 8 mm and am internal volume of
25 ± 5 ml
E 2-way solenoid valve A 2-way, 2-port solenoid valve having a minimum airflow resistance orifice
with ID > 8 mm and an opening time < 100 ms (e.g. type 256-A08, Burkert
GmbH, D-74653 Ingelfingen), or equivalent.
F Vacuum pump Pump must be capable of drawing the required flow rate through the
assembled apparatus with the powder inhaler in the mouthpiece adapter
(e.g. product type 1023, 1423 or 2565, GAST Manufacturing Inc., Benton
Harbor, MI 49022), or equivalent. Connect the pump to the 2-way solenoid
valve using short and/or wide (> 10 mm ID) vacuum tubing and connectors
to minimize pump capacity requirements.
G Timer Timer capable of driving the 2-way solenoid valve for the required time
period (e.g. type G814, RS Components International, Corby, NN17 9 RS,
UK), or equivalent.
P1 Pressure tap 2.2 mm ID, 3.1 mm OD, flush with internal surface of the sample collection
tube, centred and burr-free, 59 mm from its inlet. The pressure tap P1 must
never be open to the atmosphere.
P1 Pressure measurements Differential pressure to atmosphere (P1) or absolute pressure (P2 and P3)
P2
P3
H Flow control valve Adjustable regulating valve with maximum Cv >1, (e.g. type 8FV12LNSS,
Parker Hannifin plc., Barnstaple, EX31 1NP, UK), or equivalent.
Fig. 6: Apparatus for measuring the uniformity of delivered dose for powders for inhalation
31
Table 5 – Component specification for set-up in Fig. 7

Code Item Description
A. Connector ID > 8 mm, e.g., short metal coupling with low-diameter branch to P3.
B. Vacuum tubing A length of suitable tubing having an ID > 8 mm and an internal volume
of 25 ± 5 ml.
C. 2-way solenoid valve A 2-way, 2-port solenoid valve having a minimum airflow resistance
orifice with ID > 8 mm and an opening time < 100 ms. (e.g. type 256 -
A08), Burkert GmbH, D-74653 Ingelfingen), or equivalent.
D. Vacuum pump Pump must be capable of drawing the required flow rate through the
assembled apparatus with the powder inhaler in the mouthpiece adapter
(e.g. product type 1023, 1423 or 2565, Gast Manufacturing Inc., Benton
Harbor, MI 49022), or equivalent. Connect the pump to the 2-way
solenoid valve using short and / or wide (ID > 10 mm) vacuum tubing
and connectors to minimize pump capacity requirements.
G. Timer Timer capable to drive the 2-way solenoid valve for the required duration
(e.g. type G814, RS components International, Corby, NN17 9RS, UK),
or equivalent.
P2 Pressure measurements Determine under steady-state flow condition with an absolute pressure
transducer.
P3
F Flow control valve Adjustable regulating valve with maximum C, > 1, (e.g. type 8FV12LNSS,
Parker Hannifin plc., Barnstaple, EX311 NP, UK), or equivalent.
and let the other be open to the atmosphere. Switch on the Ensure that critical flow occurs in the flow control valve by
pump, open the 2-way solenoid valve and adjust the the following procedure; with the inhaler in place and the test
flow control valve until the pressure drop across the inhaler is flow rate Qout, measure the absolute pressure on both sides
4.0 kPa (40.8 cm H2O) as indicated by the differential pressure of the control valve (pressure reading points P2 and P3 in
meter. Remove the inhaler from the mouthpiece adapter and Figure 6). A ratio P3/P2 of less than or equal to 0.5 indicates
without touching the flow control valve, connect a flowmeter critical flow. Switch to a more powerful pump and re-measure
to the inlet of the sampling apparatus. Use a flowmeter the test flow rate if critical flow is not indicated.
calibrated for the volumetric flow leaving the meter, or calculate Predispensed systems: Prepare the inhaler as directed in the
the volumetric flow leaving the meter (Qout) using the ideal instructions to the patient and connect it to the apparatus
gas law. For a meter calibrated for the entering volumetric flow using an adapter which ensures a good seal. Draw air through
(Qin), use the following expression: the inhaler using the predetermined conditions. Repeat
the procedure until the number of deliveries which constitute
Qin × P0 the minimum recommended dose have been sampled.
Qout =
P0 × ∆P Quantitatively collect the contents of the apparatus
and determine the amount of active substance.
P0 = Atmospheric pressure.
Repeat the procedure for a further 9 doses.
DP= Pressure drop over the meter.
Reservoir systems: Prepare the inhaler as directed in the
If the flow rate is above 100 litres per minutes adjust the flow instructions to the patient and connect it to the apparatus
control valve to obtain a flow rate of 100 litres per minute (± 5 using an adapter which ensures a good seal. Draw air through
per cent). Note the volumetric airflow rate exiting the meter and the inhaler under the predetermined conditions. Repeat the
define this as the test flow rate, Qout, in litres per minute. Define procedure until the number of deliveries which constitute the
the test flow duration, T, in seconds so that a volume of 4 minimum recommended dose have been sampled.
litres of air is drawn from the mouthpiece of the inhaler at the Quantitatively collect the contents of the apparatus and
test flow rate, Qout. determine the amount of active substance.
32
Repeat the procedure for a further 2 doses. other than 28.3 litres per minute. Users must justify and validate
Discharge the device to waste until (n/2)+1 deliveries remain, the use of the impactor in the chosen conditions, when flow
where n is the number of deliveries stated on the label. If rates different from 28.3 litres per minute are selected.
necessary, store the inhaler to discharge electrostatic charges. Assemble the Andersen impactor with the pre-separator and
Collect 4 doses using the procedure described above. a suitable filter in place and ensure that the system is airtight.
Discharge the device to waste until 3 doses remain. If Depending on the product characteristics, the pre-separator
necessary, store the inhaler to discharge electrostatic charges. may be omitted, where justified and authorised. Stages 6 and
Collect 3 doses using the procedure described above. 7 may also be omitted at high flow rates, if justified. The pre-
separator may be coated in the same way as the plates or may
For preparations containing more than one active substance, contain 10 ml of a suitable solvent. Connect the apparatus to
carry out the test for uniformity of delivered dose for each a flow system according to the scheme specified in Figure 7
active substance. and Table 5.
Acceptance criteria Unless otherwise defined, conduct the test at the flow rate,
The preparation complies with the test if 9 out of 10 results lie Qout, used in the test for uniformity of delivered dose drawing
between 75 per cent and 125 per cent of the average value and 4 litres of air from the mouthpiece of the inhaler and through
all lie between 65 per cent and 135 per cent. If 2 or 3 values lie the apparatus.
outside the limits of 75 per cent to 125 per cent, repeat the test Connect a flowmeter to the induction port. Use a flowmeter
for 2 more inhalers. Not more than 3 of the 30 values lie outside calibrated for the volumetric flow leaving the meter, or calculate
the limits of 75 per cent to 125 per cent and no value lies the volumetric flow leaving the meter (Qout) using the ideal gas
outside the limits of 65 per cent to 135 per cent. law. For a meter calibrated for the entering volumetric flow
In justified and authorised cases, these ranges may be (Qin), use the following expression:
extended but no value should be greater than 150 per cent or
Qin × P0
less than 50 per cent of the average value. Qout =
P0 × ∆P
Deposition of emitted dose and fine particle dose
Apparatus.Use the apparatus described under Pressurised P0 = atmospheric pressure,
metered-dose Preparations. DP = pressure drop over the meter.
Procedure Adjust the flow control valve to achieve steady flow through
the system at the required rate, Qout (± 5 per cent). Switch off
The aerodynamic cut-off diameters of the individual stages of the pump. Ensure that critical flow occurs in the flow control
this apparatus are currently not well-established at flow rates valve by the following procedure.
Fig. 7: Experimental set-up for testing powder inhalers
33
INSULIN PREPARATIONS IP 2007
With the inhaler in place and the test flow rate established, injection into the human or animal body. They are either
measure the absolute pressure on both sides of the control solutions or suspensions or they are prepared by combining
valve (pressure reading points P2 and P3 in Figure 7). A ratio solutions and suspensions. They contain not less than 90.0
P3/P2 of less than or equal to 0.5 indicates critical flow. Switch per cent and not more than the equivalent of 110.0 per cent of
to a more powerful pump and re-measure the test flow rate if the amount of insulin stated on the label.
critical flow is not indicated.
Production
Prepare the powder inhaler for use according to the patient
instructions. With the pump running and the 2-way solenoid Insulin preparations are made by methods that are designed
valve closed, locate the mouthpiece of the inhaler in the to ensure their sterility, to avoid the introduction of foreign
mouthpiece adapter. Discharge the powder into the apparatus contaminants, bacterial endotoxins and the growth of micro-
by opening the valve for the required time, T (± 5 per cent). organisms. The methods used should confer suitable
Repeat the discharge sequence. The number of discharges properties with respect to the onset and duration of therapeutic
should be minimised and typically would not be greater than action.
10. The number of discharges is sufficient to ensure an accurate
The use of excipients in the injections may be necessary, for
and precise determination of fine particle dose.
example to make the preparation isotonic with respect to blood,
Dismantle the apparatus. Carefully remove the filter and extract to adjust the pH to the appropriate value, to prevent
the active substance into an aliquot of the solvent. Remove deterioration of the active substances or to provide adequate
the pre-separator, induction port and mouthpiece adapter from antimicrobial properties. Where appropriate, suitable
the apparatus and extract the active substance into an aliquot substances may be added and suitable procedures carried
of the solvent. Extract the active substance from the inner out to confer the appropriate physical form on the insulin-
walls and the collection plate of each of the stages of the containing component or components. Irrespective of the
apparatus into aliquots of solvent. purpose for which additives are used, they should not to
Using a suitable method of analysis, determine the quantity adversely affect the intended therapeutic action of the
of active substance contained in each of the aliquots of preparation or, at the concentration used, cause toxicity or
solvent. undue local irritation.
Calculate the fine particle dose as given under Calculations In the course of production the strength of the insulin-
for Pressurised Metered-dose Preparations. containing component or components should be determined,
where necessary, by adjustment so that the final preparation
Uniformity of Content. For dry powder inhalers in premetered contains the required number of Units of insulin per ml.
dosage units, carry out the test for uniformity of content of
the contents as given in Capsules. Initial sterilisation of the insulin-containing component or
components is done by filtration and subsequent procedures
Number of deliveries per container. Discharge doses from are carried out aseptically using materials that have been
the inhaler until empty, at the predetermined flow rate. Record sterilised by suitable methods.
the deliveries discharged. The total number of doses delivered
is not less than the number stated on the label. The final preparation is distributed aseptically into sterile glass
or plastic containers or pre-filled syringes that are closed so
Microbial contamination (2.2.9). Total viable aerobic bacterial as to exclude microbial contamination.
count. Not more than 100 cfu per g of the powder.
E. coli. Absent in 10 g of the powder. Tests
Salmonella. absent in 50 g of the powder. Insulin in the supernatant - For preparations that are
suspensions
Staphylococcus aureus. Absent in 10 g of the powder.
Not more than 2.5 per cent of the total insulin content,
Psedomonas aeruginosa. Absent in 10 g of the powder.
determined in the following manner.
Centrifuge 10 ml of the suspension for 10 minutes and carefully
separate the supernatant liquid from the residue. Determine
Insulin Preparations the insulin content of the supernatant liquid (2.3.46) and
calculate as a percentage of the total insulin content
Introduction determined as described under Assay in the individual
Insulin preparations are sterile preparations of human Insulin, monograph.
bovine Insulin or porcine Insulin intended for subcutaneous Impurities with molecular masses greater than that of insulin
34
IP 2007 INSULIN PREPARATIONS
Determine by size-exclusion chromatography (2.4.16). 17.5 minutes, insulin monomer, about 20 minutes, salts, about
Test solution. Add 4 µl of 6 M hydrochloric acid per millilitre 22 min. If the sample solution contains preservatives, for
of the preparation under examination, whether a suspension example methyl paraben, m-cresol or phenol, these compounds
or a solution, to obtain a clear acid insulin solution. When elute later. The test is not valid unless the resolution, defined
sampling a suspension, agitate the material prior to sampling by the ratio of the height of the dimer peak to the height above
in order to obtain a homogeneous sample. If a suspension the baseline of the valley separating the monomer and dimer
does not turn clear within 5 minutes of the initial addition of peaks, is at least 2.0.
hydrochloric acid, add small aliquots of acid (less than 4 µl per Inject the test solution. Record the chromatogram for
millilitre) until a solution is obtained. Preparations with approximately 35 min. In the chromatogram obtained, the sum
concentrations higher than 100 Units per ml need to be diluted of the areas of any peak with a retention time less than that of
with 0.01M hydrochloric acid to avoid overloading the the insulin peak is not greater than 3.0 per cent (protamine-
column with insulin monomer. containing preparations) or 2.0 per cent (non-protamine
containing preparations) of the total area of the peaks. Ignore
Resolution solution. Use a solution of insulin (approximately
any peak with a retention time greater than that of the insulin
4 mg per ml), containing more than 0.4 per cent of high molecular
peak.
mass proteins. An injectable insulin preparation, whether a
solution or a suspension, that has been clarified with a Related proteins
sufficient amount of 6 M hydrochloric acid, containing the
indicated percentage of high molecular mass proteins, or a Determine by liquid chromatography (2.4.14) as described
solution prepared from insulin, dissolved in 0.01 M under Assay of Insulins (2.3.46), following the elution
hydrochloric acid, may be used. Insulin containing the conditions as described in the table below:
indicated percentage of high molecular mass proteins may be Time Mobile Mobile Comment
prepared by allowing insulin powder to stand at room phase (a) phase (b)
temperature for about ten days. (min) (per cent v/v) (per cent v/v)
Maintain the solutions at 2° to 10° and use within 30 hours 0-30 42 58 isocratic
(soluble insulin injection) or 7 days (other insulin preparations). 30-44 42 →11 58 → 89 linear gradient
If an automatic injector is used, maintain the temperature at 2°
to 10°. 44-50 11 89 isocratic
Chromatographic system Maintain the solutions at 2° to 10° and use within 24 hours.
– a stainless steel column 30 cm x 7.5 mm packed with Perform a system suitability check (resolution, linearity) as
hydrophilic silica gel (5 µm to 10 µm), of a grade suitable described under Assay of Insulins (2.3.46). If necessary, the
for the separation of insulin monomer from dimers and relative proportions of the mobile phases may be adjusted to
polymers, ensure complete elution of A21 desamido porcine insulin
– mobile phase: a filtered and degassed mixture of 15 before commencement of the gradient. The profile of the
volumes of glacial acetic acid, 20 volumes of gradient may also be adjusted to ensure complete elution of
acetonitrile and 65 volumes of a 1.0 g/l solution of all insulin related impurities.
arginine,
Inject 20 µl of the test solution and 20 µl of either reference
– flow rate. 0.5 ml per minute,
solution (a), for insulin preparations containing 100 IU/ml, or
– spectrophotometer set at 276 nm,
reference solution (b), for insulin preparations containing 40
– a 100 µl loop injector.
IU/ml. If necessary, adjust the injection volume to a volume
Before using a new column for chromatographic analysis, between 10 µl and 20 µl in accordance with the results obtained
equilibrate by repeated injections of an insulin solution in the test for linearity as described under Assay. Record the
containing high molecular mass proteins. This can be done chromatograms for approximately 50 min. If necessary, make
by at least three injections of the resolution solution. The further adjustments to the mobile phase in order to ensure
column is equilibrated when repeatable results are obtained that the antimicrobial preservatives present in the test solution
from two subsequent injections. If protamine-containing are well separated from the insulin and show a shorter retention
samples are to be analysed, the equilibration of the column is time. A small reduction in the concentration of acetonitrile
performed using a solution containing protamine. increases the retention time of the insulin peaks relatively
Inject the resolution solution. When the chromatograms are more than those of the preservatives. In the chromatogram
recorded under the prescribed conditions, the retention times obtained with either reference solution (a), or reference solution
are: polymeric insulin complexes or covalent insulin-protamine (b), as appropriate, A21 desamido insulin appears as a small
complex, about 13 to 17 minutes, covalent insulin dimmer, about peak after the principal peak and has a retention time of about
35
NASAL PREPARATIONS IP 2007
1.3 relative to the principal peak, due to insulin. In the porcine insulin; (4) where applicable, that the substance is
chromatogram obtained with the test solution the area of the produced by recombinant DNA technology; (5) where
peak due to A21 desamido insulin is not greater than 5.0 per applicable, the animal species of origin; (6) the preparation
cent of the total area of the peaks; the sum of the areas of any must not be frozen; (7) where applicable, that the preparation
other peaks, apart from those due to insulin and A21 desamido must be re-suspended before use.
insulin is not greater than 6.0 per cent of the total area of the
peaks. Disregard the peaks due to the preservatives and
protamine (early eluting peaks).
Total zinc. Not more than the amount stated in the individual Nasal Preparations
monograph, determined by either of the following methods. Nasal Preparations are liquid, semi-solid or solid preparations
A. To an accurately measured volume of the gently shaken containing one or more medicaments and are intended for
injection containing 200 Units add 10 ml of alkaline borate administration to the nostrils for local or systemic effects.
buffer pH 9.0, 0.3 ml of zincon solution and sufficient water to They should as far as possible be non-irritating and should
produce 50 ml. Allow to stand for 1 hour and measure the not affect the functions of the nasal mucosa and its cilia. They
absorbance of the resulting solution at about 620 nm, using are supplied in single dose or multiple dose containers of
as the blank a solution prepared by treating 5 ml of water glass VD or plastic with, if necessary, a suitable device for
instead of the substance under examination in a similar manner. administration. They may also be supplied in pressurised
Calculate the content of zinc from the absorbance obtained containers with a suitable adaptor and with or without a
by repeating the procedure using a suitable aliquot of a mixture metering dose valve.
of 4 volumes of zinc sulphate solution and 6 volumes of water. Aqueous nasal preparations are usually isotonic and, when
B. Determine by atomic absorption spectrometry (2.4.2). supplied in multiple dose containers, contain a suitable
antimicrobial preservative except when the product itself has
Test solution. Shake the preparation gently and dilute a volume adequate antimicrobial properties.
containing 200 Units of insulin to 25.0 ml with 0.01 M
hydrochloric acid. Dilute if necessary to a suitable During manufacture, packaging, storage and distribution of
concentration of zinc (for example 0.4 µg to 1.6 µg of Zn per nasal preparations, suitable means shall be taken to ensure
millilitre) with 0.01 M hydrochloric acid. their microbial quality; acceptance criteria for microbial quality
are given in Chapter 5.9.
Reference solutions. Use solutions containing 0.40 µg, 0.80
µg, 1.00 µg, 1.20 µg and 1.60 µg of Zn per millilitre, freshly Tests
prepared by diluting zinc solution AAS (5 mg/ml Zn) with
0.01 M hydrochloric acid. Uniformity of content. Comply with the test described under
Parenteral Preparations.
Measure the absorbance at 213.9 nm using a zinc hollow-
cathode lamp as source of radiation and an air-acetylene flame Uniformity of weight. Nasal Preparations supplied in single
of suitable composition (for example 11 litres of air and 2 litres dose containers comply with the test for contents of packaged
of acetylene per minute). dosage forms (2.5.6).
Bacterial endotoxins (2.2.3). Less than 80 Units per 100 Units Nasal Drops, Solutions and Sprays
of insulin.
These are solutions, emulsions or suspensions intended for
Sterility. Comply with the test for sterility (2.2.11). instillation or spraying into the nostrils. Emulsions should
Assay. Determine as described under Assay of Insulins have a uniform appearance after shaking and should not show
((2.3.46). evidence of phase separation. Suspensions should be readily
redispersible on shaking to give a smooth and stable
Storage. Unless otherwise prescribed, store in sterile, airtight, suspension. In suspensions, the size of the dispersed particles
tamper-proof containers, protected from light, at a temperature should be such as to localise their deposition in the nostril.
of 2° to 8°. Insulin preparations should not to be frozen.
Labelling. The label states (a) the potency in Units per Nasal Powders
millilitre; (2) the concentration in terms of the number of
These are powders intended for insufflation into the nostrils
milligrams of insulin per ml (for preparations containing both
by means of a suitable device. The size of the particles should
bovine insulin and porcine insulin the concentration is stated
be such as to localise their deposition in the nostril.
as the combined amount of both insulins); (3) where applicable,
that the substance is produced by enzymatic modification of Storage. Store protected from light and moisture.
36
IP 2007 ORAL LIQUIDS
Tests Labelling. The label states (1) that the ointment is sterile,
where necessary; (2) the name and concentration of any added
Uniformity of content. Comply with the test described under antimicrobial preservative; (3) the storage conditions.
Parenteral Preparations.
Uniformity of weight. Nasal Preparations supplied in single
application containers comply with the test for contents of
packaged dosage forms (2.5.6). Oral Liquids
Oral Liquids are homogeneous liquid preparations, usually
consisting of a solution, an emulsion or a suspension of one
or more medicaments in a suitable vehicle*. They are intended
Ointments for oral administration either undiluted or after dilution. They
Ointments are homogeneous, semi-solid preparations intended may contain auxiliary substances such as suitable dispersing,
for external application to the skin or certain mucous emulsifying, suspending, wetting, solubilising, thickening,
membranes for emollient, protective, therapeutic or stabilising agents and antimicrobial preservatives. They may
prophylactic purposes where a degree of occlusion is desired. also contain suitable sweetening, flavouring and permitted
They usually consist of solutions or dispersions of one or colouring agents. if saccharin, including its sodium and
more medicaments in suitable bases. They are formulated using potassium salts, is used as a sweetening agent, its
hydrophobic, hydrophillic or water-emulsifying bases to concentration in preparations meant for paediatric use should
provide preparations that are immiscible, miscible or be restricted so as to limit its intake to 5 mg per kg of body
emulsifiable with the skin secretion, respectively. The base weight.
should not produce irritation or sensitisation of the skin, nor Oral Liquids other than Oral Emulsions may be supplied as
should it retard wound healing; it should be smooth, inert, liquids or prepared just before use by dissolving or dispersing
odourless or almost odourless, physically and chemically granules or powder in the liquid stated on the label. The
stable and compatible with the skin and with incorporated granules or powder comply with the requirements stated under
medicaments. The proportions of the base ingredients should Oral Powders.
be such that the ointment is not too soft or too hard for
convenient use. The consistency should be such that the
oral liquids, suitable means shall be taken to ensure their
ointment spreads and softens when stress is applied.
microbial quality; acceptance criteria for microbial quality are
Ointments may contain suitable auxiliary substances such as given in Chapter 5.9.
antioxidants, stabilisers, thickeners and emulsifiers and, when
Oral Liquids should not be diluted and stored; where, however,
the base might support the growth of microbial contaminants,
the individual monograph directs dilution, the diluted Oral
suitable antimicrobial preservatives.
Liquid should be freshly prepared irrespective of the nature
During manufacture, packaging, storage and distribution of of the diluent. Diluted Oral Liquids may be less stable
ointments, suitable means shall be taken to ensure their physically and chemically than the corresponding undiluted
microbial quality; acceptance criteria for microbial quality are preparation and should be used within the period stated on
given in Chapter 5.9. the label.
If an ointment is specifically intended for use on large wounds Oral Liquids are variously known as Elixirs, Linctuses Mixtures,
or on severely injured skin it should be sterile. Oral Drops, Oral Emulsions, Oral Solutions, Oral Suspensions
Ointments should not normally be diluted; if dilution is and Syrups. These terms are defined below.
necessary care should be taken to choose the right diluent to Elixirs. Elixirs are clear, flavoured Oral Liquids containing one
avoid risk of instability or incompatibility. or more active ingredients dissolved in a vehicle that usually
contains a high proportion of Sucrose or a suitable polyhydric
Tests alcohol or alcohols and may also contain Ethanol (95 per cent)
or a dilute Ethanol.
Uniformity of weight. Comply with the test for contents of
packaged dosage forms (2.5.6). Linctuses. Linctuses are viscous Oral Liquids containing one
or more active ingredients dissolved in a vehicle that usually
Sterility. When the ointment is labelled as sterile, it complies
contains a high proportion of sucrose, other sugars or a
with the test for sterility (2.2.11).
suitable polyhydric alcohol or alcohols. Linctuses are intended
Storage. Store at a temperature not exceeding 30º unless for use in the treatment or relief of cough, and are sipped and
otherwise directed. Do not freeze. swallowed slowly without the addition of water.
37
ORAL LIQUIDS IP 2007
Mixtures. Mixtures are Oral Liquids containing one or more active ingredient, carry out the test for each active ingredient
active ingredients dissolved, suspended or dispersed in a that corresponds to the above conditions. Empty each
suitable vehicle. Suspended solids may separate slowly on container as completely as possible and carry out the test on
keeping but are easily redispersed on shaking. the individual contents of active ingredients.
Oral Drops. Oral Drops are Oral Liquids that are intended to The test for Uniformity of content should be carried out only
be administered in small volumes with the aid of a suitable after the content of active ingredient(s) in a pooled sample of
measuring device such as a dropper. the preparation has been shown to be within the accepted
limits of the stated content.
Oral Emulsions. Oral Emulsions are Oral Liquids containing
one or more active ingredients and are stabilised oil-in-water Determine the content of active ingredient(s) of each of 10
dispersions, either or both phases of which may contain containers taken at random using the method given in the
dissolved solids. Solids may also be suspended in Oral monograph or by any other suitable analytical method of
Emulsions. Emulsions may exhibit phase separation but are equivalent accuracy and precision. The preparation complies
easily reformed on shaking. The preparation remains with the test if the individual values thus obtained are all
sufficiently stable to permit a homogeneous dose to be between 85 to 115 per cent of the average value. The
withdrawn. preparation fails to comply with the test if more than one
individual value is outside the limits 85 to 115 per cent of the
Oral Solutions. Oral Solutions are Oral Liquids containing
average value or if any one individual value is outside the
one or more active ingredients dissolved in a suitable vehicle.
limits 75 to 125 per cent of the average value. If one individual
Oral Suspensions. Oral Suspensions are Oral Liquids value is outside the limits 85 to 115 per cent but within the
containing one or more active ingredients suspended in a limits 75 to 125 per cent of the average value, repeat the
suitable vehicle. Suspended solids may slowly separate on determination using another 20 containers taken at random.
keeping but are easily redispersed. The preparation complies with the test if in the total sample of
In the manufacture of oral suspensions containing dispersed 30 containers not more than 3 individual values are outside
particles, measures shall be taken to ensure a suitable and the limits 85 to 115 per cent and not more than one is outside
controlled particle size with regard to the intended use of the the limits 75 to 125 per cent of the average value.
product. Uniformity of weight/volume. Unless otherwise specified, Oral
Syrups. Syrups are viscous Oral Liquids that may contain Liquids comply with the test for contents of packaged dosage
one or more active ingredients in solution. The vehicle usually forms (2.5.6).
contains large amounts of Sucrose or other sugars to which Storage. Store Oral Liquids or powders and granules for the
certain polyhydric alcohols may be added to inhibit preparation of Oral Liquids in well-closed containers at
crystallisation or to modify solubilisation, taste and other temperatures not exceeding 30º.
vehicle properties. Sugarless syrups may contain sweetening
Labelling. For Oral Liquids that are supplied as drops, the
agents and thickening agents. Syrups may contain Ethanol
label states the number of drops per g of preparation if the
(95%) as a preservative or as a solvent to incorporate
dose is stated in drops or the number of drops per ml of
flavouring agents. Antimicrobial agents may also be added to
preparation if the dose is stated in volume. For oral liquids
Syrups.
supplied as granules or powder to be constituted before use,
Containers. Oral Liquids may be supplied in multiple dose or the label states (1) that the contents are meant for preparation
single dose containers. Oral Emulsions and Oral Suspensions of an Oral Liquid; (2) the directions for preparing the Oral
should be packed in bottles sufficiently wide-mouthed to liquid including the nature and quantity of the liquid to be
facilitate the flow of the contents. They are administered either used; (3) the conditions under which the constituted solution
in volumes such as 5 ml, or multiples of 5 ml, or in small volumes should be stored; (4) the period during which the constituted
(drops). Each dose of a multiple dose Oral Liquid is Oral Liquid may be expected to remain satisfactory for use
administered by means of a suitable measuring device which when prepared and stored in accordance with the
is usually provided with the container. manufacturer’s recommendations; (5) the strength in terms of
the active ingredient(s) in a suitable dose-volume of the
Tests constituted preparation.
Uniformity of content. Unless otherwise specified, single dose
liquids in suspension form or powders or granules presented * The term vehicle means a carrier, composed of one or more
in single dose containers and that contain less than 10 mg or excipients, for the active pharmaceutical ingredient(s) in a liquid
less than 10 per cent of active ingredient comply with the preparation.
following test. For Oral Liquids containing more than one
38
IP 2007 PARENTERAL PREPARATIONS
Oral Powders determination using another 20 containers taken at random.

The preparation complies with the test if in the total sample of
Oral Powders are finely divided powders that contain one or 30 containers not more than 3 individual values are outside
more medicaments with or without auxilliary substances the limits 85 to 115 per cent and not more than one is outside
including, where specified, flavouring and colouring agents. the limits 75 to 125 per cent of the average value.
However, addition of saccharin or its salts is not permitted in
the preparations meant for paediatric use. They are intended NOTE — The test for Uniformity of content is not applicable
to be taken internally with or without the aid of water or any to preparations containing multivitamins and trace elements.
other suitable liquid. Uniformity of weight. Unless otherwise specified, Oral
Oral Powders may be single dose or multiple dose preparations. Powders presented in single dose containers comply with the
For single dose powders, each dose is enclosed in a separate test for contents of packaged dosage forms (2.5.6).
container, e.g., a sachet, a paper packet or a vial. With multiple
dose powders it may be necessary to provide a measuring
device capable of delivering the quantity prescribed.
Parenteral Preparations
Effervescent Oral Powders are intended to be dissolved or
dispersed in water before administration. Injectable Preparations
In the manufacture of oral powders, means are taken to ensure NOTE — The provisions of this monograph do not necessarily
a suitable particle size with regard to the intended use of the apply to Blood Products or Immunological Products because
product. During manufacture, packaging, storage and of their special nature and licensing requirements.
distribution of oral powders, suitable means shall be taken to
ensure their microbial quality; acceptance criteria for microbial Introduction
quality are given in Chapter 5.9.
Parenteral Preparations are sterile products intended for
Storage. Store Oral Powders in containers protected from administration by injection, infusion or implantation into the
moisture. body. They may be preparations intended for direct parenteral
administration or they may be parenteral products for
Tests constituting or diluting prior to administration. There are five
Uniformity of content. Unless otherwise specified, Oral main types of Parenteral Preparations, namely, Injections,
Powders presented in single dose containers that contain less Infusions, Powders for Injection, Concentrated Solutions for
than 10 mg of active ingredient per dose or that contain less Injection and Implants.
than 10 per cent w/w of active ingredient comply with the
following test. For Oral Powders containing more than one Production
active ingredient carry out the test for each active ingredient Parenteral Preparations should be prepared by methods
that corresponds to the above conditions. Empty each designed to ensure their sterility and to avoid the introduction
container as completely as possible and carry out the test on of foreign contaminants, the presence of pyrogens or of
the individual contents of active ingredients. bacterial endotoxins and the growth of micro-organisms.
The test for Uniformity of content should be carried out only Parenteral Preparations which are solutions or suspensions
after the content of active ingredient(s) in a pooled sample of require vehicles in which the medicaments are incorporated.
the preparation has been shown to be within the accepted The most commonly used vehicle is Water for Injections that
limits of the stated content. complies with the requirements for water for injections in bulk
Determine the content of active ingredient(s) of each of 10 stated in the monograph on Water for injections. Any other
containers taken at random using the method given in the suitable vehicles may be used provided they are safe in the
monograph or by any other suitable analytical method of volume of injections administered and also do not interfere
equivalent accuracy and precision. The preparation complies with the therapeutic efficacy of the preparation or with its
with the test if the individual values thus obtained are all response to the prescribed tests and assays of the
between 85 to 115 per cent of the average value. The Pharmacopoeia. It may be necessary to include auxiliary
preparation fails to comply with the test if more than one substances to increase the stability or usefulness of the
individual value is outside the limits 85 to 115 per cent of the preparation, unless otherwise specified in the individual
average value or if any one individual value is outside the monograph. Such substances at the concentration at which
limits 75 to 125 per cent of the average value. If one individual they are used should not adversely affect the intended
value is out-side the limits 85 to 115 per cent but within the medicinal action of the preparation nor cause toxicity or local
limits 75 to 125 per cent of the average value, repeat the irritation and should not interfere with the responses to the
39
PARENTERAL PREPARATIONS IP 2007
specified tests and assays. No colouring agent may be added nominal dose using normal technique. They must be used for
solely for the purpose of colouring the finished preparation. all parenteral preparations administered at one time in volumes
Aqueous Parenteral Preparations for administration by the of 10 ml or more.
subcutaneous, intradermal, intramuscular, or in the case of Multiple dose containers permit the withdrawal of successive
large volumes, intravenous route, should if possible be made portions of the contents without removal or destruction of
isotonic with blood by the addition of Sodium Chloride or the closure and without changing the strength, quality or
other suitable substances. Buffering agents should not be purity of the remaining portion. They may be used for
used in preparations intended for intraocular or intracardiac intramuscular, subcutaneous or intracutaneous administration,
injection, or in products that may gain access to the but no multiple dose container may contain a total volume of
cerebrospinal fluid. injection sufficient to permit the withdrawal of more than ten
Parenteral Preparations that are packaged in multiple dose doses, unless otherwise stated in the individual monograph.
containers, regardless of the method of sterilisation employed, The period of time between the withdrawal of the first and
may contain suitable antimicrobial preservatives in appropriate final dose should not be unduly prolonged.
concentration, unless otherwise directed in the individual A multiple dose container for a sterile solid permits the addition
monograph, or unless the active ingredients themselves are of a suitable vehicle and withdrawal of portions of the resulting
bacteriostatic. The effectiveness of the chosen preservative preparation in such a manner that the sterility of the product
shall have been demonstrated during the development of a is maintained.
parenteral preparation.
Closures. Vials or bottles are fitted with suitable closures that
Precautions to be taken for administration and for storage ensure a good seal, prevent the access of micro-organisms
between successive withdrawls from such multiple dose and other contaminants and usually permit the withdrawal of
preparations should be indicated. Preservatives should not a part or the whole of the contents of the container without
be added when the volume to be injected as a single dose removal of the closure. The plastic or rubber materials of which
exceeds 15 ml, unless otherwise justified, or when the the closure is composed must be compatible with the
preparation is intended for administration by the intraocular, preparation and be sufficiently firm and elastic to allow the
intracardiac or intracisternal routes (or other route giving passage of a needle with minimal shedding of particles and to
access to the cerebrospinal fluid). ensure that the puncture is resealed when the needle is
Where the active ingredient is susceptible to oxidative withdrawn. Requirements concerning closures are given in
degradation a suitable antioxidant may be added and/or the Chapter 6.3.
air in the container may be evacuated or displaced by oxygen- Before use, closures should be washed with a suitable
free nitrogen or other suitable inert gas. detergent and rinsed with and boiled in several changes of
Sterilisation. Methods of sterilisation that may be used in the Purified Water. Closures made from rubber and synthetic
manufacture of Parenteral Preparations are described in materials are liable to absorb the ingredients of the parenteral
Chapter 5.3. preparation with which they are used, e.g., the preservative.
Containers. Containers for Parenteral Preparations are made When an antimicrobial preservative is used the closure, when
as far as possible from materials that (1) are sufficiently necessary, should be placed in a solution of that preservative
transparent to permit visual inspection of the contents, except in Purified Water containing at least twice the concentration
for implants; (2) do not adversely affect the quality of the to be used in the preparation; the quantity of solution used
preparation under the ordinary conditions of handling, should be sufficient to cover the closures and should be at
shipment, storage, sale and use; (3) do not permit diffusion least 2 ml for each g of the material. The vessel should then be
into or across the walls of the container or yield foreign closed and heated at an appropriate combination of time and
substances into the preparation. Parenteral Preparations may temperature. After heating, the closures should be kept in the
be supplied in glass ampoules, vials or bottles or in other sealed container until required for use.
containers such as plastic bottles or bags or in prefilled When the parenteral preparation with which the closures are
syringes the integrity of which is ensured by suitable means. to be used contains other added substances that are liable to
Requirements concerning containers are given in Chapter 6.2. be absorbed by the closure, these should be added to the
Single dose containers are used for administration of the solution in which the closures are to be heated in amounts
contents on one occasion only and are to be preferred for all equal to at least twice the concentration to be used in the
parenteral preparations. They may be used for intrathecal, parenteral preparation. Closures intended for containers of
intracardiac, intracisternal or intravenous injectable oily preparations should be made of oil-resistant materials.
preparations. They contain sufficient of the Parenteral Inspection. Good Manufacturing Practices require that each
Preparation to permit the withdrawal and administration of the final container of a Parenteral Preparation be subjected
40
IP 2007 PARENTERAL PREPARATIONS
individually to a physical inspection whenever the nature of Uniformity of content. Unless otherwise stated in the individual
the container permits and that every container the contents of monograph, suspensions for injection that are presented in
which show evidence of contamination with visible foreign single dose containers and that contain less than 10 mg or
material be rejected. less than 10 per cent of active ingredient comply with the
following test. For suspensions for injection containing more
Labelling. Containers of Parenteral Preparations should be
than one active ingredient carry out the test for each active
labelled in a manner that sufficient area of the container remains
ingredient that corresponds to the above conditions.
uncovered for its full length or circumference to permit
inspection of the contents. The label of a Parenteral Preparation The test for Uniformity of content should be carried out only
states (1) the name of the Parenteral Preparation; (2) the strength after the content of active ingredient(s) in a pooled sample of
in terms of the amount of active ingredient in percentage or in the preparation has been shown to be within accepted limits
a suitable dose-volume; (3) the name and proportion of or of the stated content.
antimicrobial preservative added; (4) the conditions under Determine the content of active ingredient(s) of each of 10
which the preparation should be stored. containers taken at random, using the method given in the
In the case of Parenteral Preparations like Powders for Injection monograph or by any other suitable analytical method of
and Concentrated Solutions for Injection wherein a diluent is equivalent accuracy and precision. The preparation under
intended to be added before use, the label also states (1) the examination complies with the test if the individual values
composition of the recommended diluent; (2) the conditions thus obtained are all between 85 and 115 per cent of the average
under which the constituted preparation should be stored; (3) value. The preparation under examination fails to comply with
the period within which the constituted solution should be the test if more than one individual value is outside the limits
used if it has been stored under the recommended conditions 85 to 115 per cent of the average value or if any one individual
of storage after constitution. In the case of Powders for value is outside the limits 75 to 125 per cent of the average
Injection, the label also states the amount of diluent to be value. If one individual value is outside the limits 85 to 115 per
used to attain a specific concentration of the active ingredient cent but within the limits 75 to 125 per cent of the average
in the solution or suspension so obtained whereas in the case value, repeat the determination using another 20 containers
of Concentrated Solutions for Injection, the amount of diluent taken at random. The preparation under examination complies
to be used to attain a specific concentration and the final with the test if in the total sample of 30 containers not more
volume of the solution or suspension so obtained. than one individual value is outside the limits 85 to 115 per
cent and none is outside the limits 75 to 125 per cent of the
average value.
Injections
NOTE — The test for Uniformity of content is not applicable
Injections are sterile solutions, emulsions or suspensions. to suspensions for injection containing multivitamins and
They are prepared by dissolving, emulsifying or suspending trace elements.
the active ingredient(s) and any added substances in Water
for Injection or in a suitable non-aqueous vehicle, or in a Extractable volume. Where the nominal volume does not
mixture of the two if they are miscible. exceed 5 ml, the containers comply with the requirements of
Method 1 and where the nominal volume is greater than 5 ml,
Injections that are emulsions should not show any evidence the containers comply with the requirements of Method 2.
of separation and show a uniform appearance after shaking. Suspensions should be shaken before the contents are
The diameter of the globules of the dispersed phase of withdrawn; oily injections may be warmed but should be cooled
emulsions intended for intravenous injection must be decided to 25º before carrying out the test.
with regard to the use of the preparation. Injections that are
Method 1 — Use 6 containers, 5 for the test and 1 for rinsing
suspensions may show a sediment which is readily dispersible
the syringe used. Inspect the 5 containers to be used in the
on shaking. The suspension remains sufficiently stable to
test visually and ensure that each contains approximately the
enable a homogenous dose to be withdrawn from the container.
same volume of the preparation.
Tests Using a syringe with a capacity not exceeding twice the volume
to be measured and fitted with a suitable needle, take up a
Particulate matter. Injections that are solutions, when small quantity of the liquid under examination from the
examined under suitable conditions of visibility, are clear and container reserved for rinsing the syringe, and discharge it
practically free from particles that can be observed on visual from the syringe whilst the needle is pointing upwards so as
inspection by the unaided eye. Injections that are supplied in to expel any air. Withdraw as much as possible the contents of
containers with a nominal content of 100 ml or more comply one of the containers reserved for the test and transfer, without
with the test for particulate contamination (2.5.9). emptying the needle, to a dry graduated cylinder of such
41
PARENTERAL PREPARATIONS IP 2007
capacity that the total combined volume to be measured Powders for injection
occupies not less than 40 per cent of the nominal volume of
the cylinder. Repeat the procedure until the contents of the 5 Powders for injection are sterile, solid substances (including
containers have been transferred and measure the volume. freeze-dried materials) which are distributed in their final
The average content of the 5 containers is not less than the containers and which, when shaken with the prescribed volume
nominal volume and not more than 115 per cent of the nominal of the appropriate sterile liquid, rapidly form clear and
volume. practically particle-free solutions or uniform suspensions.
Method 2 — Transfer the contents of not less than 3 containers Tests
separately to dry graduated cylinders such that the volume to
Powders for injection comply with the requirements of tests
be measured occupies not less than 40 per cent of the nominal
stated under individual monographs and with the following
volume of the cylinder and measure the volume transferred.
requirements.
The contents of each container are not less than the nominal
volume and not more than 110 per cent of the nominal volume. Uniformity of content. Unless otherwise stated in the individual
Multiple dose containers labelled to yield a specific number monograph, Powders for injection that contain 10 mg or less
of doses shall contain a sufficient excess to permit the than 10 mg or less than 10 per cent of active ingredient or that
withdrawal of the designated number of doses. have a unit weight equal to or less than 50 mg comply with the
test for Uniformity of content described under Injections. For
Sterility (2.2.11). Injections comply with the test for sterility. Powders for injection containing more than one active
Pyrogens. Unless otherwise stated in the individual ingredient carry out the test for each active ingredient that
monograph, when the volume to be injected in a single dose is corresponds to the above conditions. The test is not applicable
10 ml or more, Injections comply with the test for pyrogens to Powders for injection containing multivitamins and trace
(2.2.8), unless the test for bacterial endotoxins (2.2.3), is elements.
prescribed. The test for Uniformity of content should be carried out only
after the content of active ingredient(s) in a pooled sample of
Infusions the preparation has been shown to be within accepted limits
Infusions are sterile aqueous solutions or emulsions with water of the stated content.
as the continuous phase. They are free from pyrogens or Uniformity of weight. For Powders for injection that are
bacterial endotoxins, are usually made isotonic with blood required to comply with the test for Uniformity of content of
and do not contain any added antimicrobial preservatives. all active ingredients, the test for Uniformity of weight is not
Intravenous Infusions that are emulsions do not show any required.
evidence of phase separation. The diameter of the globules of
Remove any adherent labels from a container and wash and
the dispersed phase of emulsions must be decided with regard
dry the outside. Open the container and immediately weigh
to the use of the preparation.
the container and its contents. Empty the container as
Tests completely as possible by gentle tapping, rinse if necessary
with water and then with ethanol (95 per cent) and dry at 100º
Intravenous Infusions comply with the requirements of tests to 105º for 1 hour or, if the nature of the container precludes
stated under individual monographs and with the following such treatment, dry at a lower temperature to constant weight.
requirements. Allow to cool in a desiccator and weigh. The difference
Particulate contamination. Intravenous Infusions that are between the weights represents the weight of the contents.
solutions, when examined under suitable conditions of visibility, Repeat the procedure with a further 19 containers and
are clear and practically free from particles that can be observed determine the average weight. Not more than two of the
on visual inspection by the unaided eye. Intravenous individual weights deviate from the average weight by more
Infusions that are solutions and are supplied in containers than 10 per cent and none deviates by more than 20 per cent.
with a nominal content of 100 ml or more comply with the test
Clarity of solution. Constitute the injection as directed on the
for particulate contamination (2.5.9).
label.
Sterility (2.2.11). Intravenous Infusions comply with the test a) The solid dissolves completely, leaving no visible residue
for sterility. as undissolved matter.
Pyrogens. Where no test for bacterial endotoxins (2.2.3) is b) The constituted injection is not significantly less clear
prescribed, Intravenous Infusions comply with the test for than an equal volume of the diluent or of water for
pyrogens (2.2.8). Unless otherwise stated in the individual injections contained in a similar container and examined
monograph inject 10 ml per kg of body weight into each animal. in the same manner.
42
IP 2007 PESSARIES
Particulate matter. Constitute the injection as directed on the cooling in order to solidify the mass. Auxiliary substances
label; the solution is essentially free from particles of foreign normally used are mixtures of mono-, di- and triglycerides of
matter that can be seen on visual inspection. saturated fatty acids, macrogols, theobroma oil and gelatinous
Sterility (2.2.11). Powders for injection comply with the test mixtures consisting of Gelatin, Glycerin and Water.
for sterility. Moulded Pessaries are smooth and are usually ovoid in shape
but may also be of various other shapes and of various
Concentrated Solutions for injection volumes. When examined microscopically, their surfaces and
longitudinal sections are normally of uniform texture except
Concentrated Solutions for injection are sterile solutions that
where the pessary consists of many layers.
are intended to be administered by injection or by intravenous
infusion only after dilution with a suitable liquid. Storage. Store in ventilated containers.
Tests Shell Pessaries. Shell Pessaries, also known as Vaginal

Capsules, are similar to Soft Capsules, differing only in their
After dilution Concentrated Solutions for injection comply shape and size. They are commonly ovoid in shape, smooth
with the requirements of tests for Injections or Infusions as and have a uniform appearance.
appropriate. Storage. Store in well-closed containers.
Implants
Tests
Implants are sterile solid preparations of size and shape suitable Uniformity of container contents. Comply with the test for
for implantation into body tissues so as to release the active contents of packaged dosage forms (2.5.6).
ingredient over an extended period of time. They are normally
presented individually in sterile containers. Uniformity of content. The test is applicable to Pessaries that
contain less than 10 mg or less than 10 per cent of active
Tests ingredient. For Pessaries containing more than one active
ingredient carry out the test for each active ingredient that
Sterility (2.2.11). Implants comply with the test for sterility. corresponds to the above conditions.
The test for Uniformity of content should be carried out only
after the content of active ingredient(s) in a pooled sample of
Pessaries the pessaries has been shown to be within accepted limits of
the stated content.
Pessaries are solid preparations containing one or more active
ingredients and are suitable for vaginal insertion. They are Carry out the test for Uniformity of content described under
normally intended for use as a single dose. Capsules.
The active ingredients are dissolved or dispersed in a suitable Uniformity of weight. This test is not applicable to Pessaries
basis containing one or more auxiliary substances that may that are required to comply with the test for Uniformity of
be dispersible, soluble or insoluble in water. The auxiliary content for all active ingredients.
substances may be similar to the ones used for Suppositories Weigh individually 20 pessaries, taken at random, and
or Tablets; such substances must be innocuous and determine the average weight. Not more than two of the
therapeutically inert in the quantities present. individual weights deviate from the average weight by more
During manufacture, packaging, storage and distribution of than 5 per cent and none deviates by more than 10 per cent.
pessaries, suitable means shall be taken to ensure their Disintegration. This test is not necessarily applicable to
microbial quality; acceptance criteria for microbial quality are Pessaries intended for modified release or for prolonged
given in Chapter 5.9. local action.
Compressed Pessaries. Compressed Pessaries, also known Carry out the disintegration test (2.5.1). Disintegration occurs
as Vaginal Tablets, have the general characteristics of Uncoated in not more than 30 minutes for Compressed Pessaries and
Tablets but are usually large and of greater weight. Shell Pessaries and in not more than 60 minutes for Moulded
Storage. Store in well-closed containers, protected from Pessaries.
moisture and from being crushed.
Suppositories
Moulded Pessaries. Moulded Pessaries are manufactured by
pouring the liquefied mass containing the medicament(s) and Suppositories are solid preparations each containing one or
auxiliary substances into moulds of suitable volume and more active ingredients and are suitable for rectal
43
TABLETS IP 2007
administration. They are normally intended for use as a single Because of their composition, method of manufacture or
dose for local action or systemic absorption of the active intended use, tablets present a variety of characteristics and
ingredients. consequently there are several categories of tablets.
The active ingredients are ground and passed through a sieve, Unless otherwise stated in the individual monograph, tablets
if necessary, and dissolved or dispersed in a suitable basis are uncoated. Where coating is permitted, the monograph
that may be soluble or dispersible in water or that may melt at states “The tablets may be coated”. Where the monograph
body temperature. directs coating the statement reads “The tablets are coated”.
Unless otherwise directed, tablets may be coated in one of
Suppositories may contain suitable auxiliary substances such
different ways.
as adsorbents, diluents, lubricants, antimicrobial preservatives
and colouring agents permitted under the Drugs and Cosmetics Tablets are usually solid, right circular cylinders, the end
Rules, 1945. surfaces of which are flat or convex and the edges of which
may be bevelled. They may exist in other shapes like triangular,
Moulded Suppositories. Moulded Suppositories are
rectangular, etc also. They may have lines or break-marks and
manufactured by liquefying by heating the mass containing
may bear a symbol or other markings. They are sufficiently
the medicament(s) and auxiliary substances and then pouring
hard to withstand handling without crumbling or breaking.
the mass into moulds of suitable volume and cooling in order
to solidify the mass. In some cases, the solid medicated mass Production
may be cold-moulded by compression in a suitable matrix.
Tablets are obtained by compression of uniform volumes of
Moulded Suppositories have the characteristics of Moulded
powders or granules by applying high pressures and using
Pessaries.
punches and dies. The particles to be compressed consist of
Shell Suppositories. Shell Suppositories, also known as Rectal one or more medicaments, with or without auxiliary substances
Capsules, are generally similar to Soft Capsules except that such as diluents, binders, disintegrating agents, lubricants,
they may have lubricating coatings. glidants, permitted colours and substances capable of
Shell Suppositories have the characteristics of Shell Pessaries. modifying the behaviour of the medicaments in the digestive
tract. Such substances must be innocuous and therapeutically
During manufacture, packaging, storage and distribution of inert in the quantities present.
suppositories, suitable means shall be taken to ensure their
microbial quality; acceptance criteria for microbial quality are In the production of tablets, measures are taken to ensure that
given in Chapter 5.9. they have sufficient strength to avoid crumbling or breaking
on handling or subsequent handling. Chewing tablets are
Tests manufactured to ensure that they are easily crushed by
chewing.
Moulded Suppositories and Shell Suppositories comply with
the tests stated under Moulded Pessaries and Shell Pessaries
tablets, suitable means shall be taken to ensure their microbial
respectively.
Storage. Store in well-closed containers. Chapter 5.9.
Tests
Tablets NOTE — Unless otherwise stated below or in the individual
monograph, the following tests apply to all categories of
NOTE — The provisions of this monograph do not necessarily
tablets.
apply to tablets intended for use other than by oral
administration such as Vaginal preparations or Oromucosal Uniformity of container contents. Tablets comply with the
preparations, and to lozenges, oral pastes and oral gums. test for contents of packaged dosage forms (2.5.6).
Content of active ingredients. Determine the amount of active
Introduction
ingredient(s) by the method described in the Assay and
Tablets are solid dosage forms each containing a unit dose of calculate the amount of active ingredient(s) per tablet. The
one or more medicaments. They are intended for oral result lies within the range for the content of active
administration. Some tablets are swallowed whole or after being ingredient(s) stated in the monograph. This range is based on
chewed, some are dissolved or dispersed in water before the requirement that 20 tablets, or such other number as may
administration and some are retained in the mouth where the be indicated in the monograph, are used in the Assay. Where
active ingredient is liberated. 20 tablets cannot be obtained, a smaller number, which must
44
IP 2007 TABLETS
not be less than 5, may be used, but to allow for sampling individual monograph. Uncoated Tablets have the general
errors the tolerances are widened in accordance with Table 1. characteristics of tablets. When a broken section of an
The requirements of Table 1 apply when the stated limits are uncoated tablet is examined under a lens, either a relatively
between 90 and 110 per cent. For limits other than 90 to 110 per uniform texture (single-layer tablets) or a stratified structure
cent, proportionately smaller or larger allowances should be (multi-layer tablets) is seen; there are no signs of coating.
made.
Table 1
Tests
Disintegration (2.5.1). Use water as the liquid. Add a disc to
Weight of active Subtract from Add to the upper
each tube. Operate the apparatus for 15 minutes, unless
ingredients in each lower limit limit for
otherwise stated in the individual monograph. Examine the
tablet for samples of samples of
state of the tablets. If the tablets fail to comply because of
15 10 5 15 10 5 adherence to the discs, repeat the test on a further 6 tablets
0.12 g or less 0.2 0.7 1.6 0.3 0.8 1.8 omitting the discs. The tablets comply with the test if all 6
tablets have disintegrated.
More than 0.12 g 0.2 0.5 1.2 0.3 0.6 1.5
but less than 0.3 g The test does not apply to chewable tablets.
0.3 g or more 0.1 0.2 0.8 0.2 0.4 1.0
Coated Tablets
Uniformity of content (2.5.4). This test is applicable to tablets
Coated tablets are tablets covered with one or more layers of
that contain 10 mg or less than 10 mg or less than 10 per cent
mixtures of various substances such as resins, gums, inactive
w/w of active ingredient. For tablets containing more than
and insoluble fillers, sugars, plasticisers, polyhydric alcohols,
one active ingredient carry out the test for each active
waxes, etc. The coating may also contain medicaments. In
ingredient that corresponds to the aforementioned conditions.
compression-coated tablets, the coating is applied by
The test is also applicable to coated tablets other than film-
compressing around the tablets granules prepared from tablet
coated tablets, irrespective of their content of active
excipients such as lactose, calcium phosphate, etc. Substances
substance(s).
used as coatings are usually applied as a solution or
The test for Uniformity of content should be carried out only suspension in conditions in which evaporation of the vehicle
after the content of active ingredient(s) in a pooled sample of occurs. When the coating is thin, the tablets are described as
the tablets has been shown to be within accepted limits of the film-coated.
stated content.
Coated tablets may contain flavouring agents.
The test for Uniformity of content is not applicable to tablets
Coated tablets have a smooth, usually polished and often
containing multivitamins and trace elements.
coloured, surface; a broken section examined under a lens
Uniformity of weight (2.5.3). This test is not applicable to shows a core surrounded by one or more continuous layers
coated tablets other than film-coated tablets and to tablets of a different texture.
that are required to comply with the test for uniformity of
content for all active ingredients. Tests
Dissolution (2.5.2). Where required, the requirements for this Disintegration (2.5.1). For coated tablets other than film-
test are given in the individual monographs. Where a coated tablets.
dissolution test is prescribed, the disintegration test may not
be necessary. Use water as the liquid. Add a disc to each tube. Operate the
apparatus for 60 minutes, unless otherwise stated in the
Uncoated Tablets individual monograph. Examine the state of the tablets. If any
of the tablets has not disintegrated, repeat the test on a further
Uncoated tablets may be single-layer tablets resulting from a 6 tablets, replacing water with 0.1 M hydrochloric acid. The
single compression of particles or multi-layer tablets consisting tablets comply with the test if all 6 tablets have disintegrated
of parallel layers obtained by successive compression of in the acid medium.
particles of different compositions. No treatment is applied to
such tablets after compression. Any added substances are
For film-coated tablets.
not specifically intended to modify the release of their active
ingredient(s) in the digestive fluids. Carry out the test described above but operate the apparatus
The addition of flavouring agents to uncoated tablets other for 30 minutes, unless otherwise stated in the individual
than multi-layer tablets is not official unless permitted in the monograph.
45
TABLETS IP 2007
If coated tablets fail to comply because of adherence to the Enteric-coated Tablets

discs, repeat the test on a further 6 tablets omitting the discs.
The tablets comply with the test if all 6 tablets have Enteric-coated tablets (Gastro-resistant tablets) are delayed-
disintegrated. release tablets that are intended to resist the gastric fluid but
to release their active ingredient(s) in the intestinal fluid. For
The test does not apply to chewable tablets. this purpose substances such as cellulose acetate phthalate
and anionic copolymers of methacrylic acid and its ethers are
Dispersible Tablets used for providing tablets with a gastric-resistant coating or
for covering either granules or particles with gastric-resistant
Dispersible tablets are uncoated or film-coated tablets that
coating.
produce a uniform dispersion in water and may contain
permitted flavouring and sweetening agents. However, if Enteric-coated tablets have the characteristics of Coated
saccharin, including its sodium and potassium salts, is used Tablets.
as a sweetening agent, its concentration in dispersible tablets
meant for paediatric use should be restricted so as to limit its Tests
intake to 5 mg/kg of body weight. Disintegration (2.5.1). If the tablet has a soluble external
coating, immerse the basket in water at room temperature for
Tests 5 minutes. Suspend the assembly in the beaker containing 0.1
Disintegration (2.5.1). Determine at 24º to 26º and operate the M hydrochloric acid and operate without the discs for 120
apparatus for 3 minutes. minutes, unless otherwise stated in the individual monograph.
Remove the assembly from the liquid. No tablet shows signs
Uniformity of dispersion. Place 2 tablets in 100 ml of water
of cracks that would allow the escape of the contents of
and stir gently until completely dispersed. A smooth
disintegration, apart from fragments of coating. Replace the
dispersion is obtained which passes through a sieve screen
liquid in the beaker with mixed phosphate buffer pH 6.8, add
with a nominal mesh aperture of 710 mm (sieve number 22).
a disc to each tube and operate the apparatus for a further 60
minutes. Remove the assembly from the liquid. The tablets
Effervescent Tablets pass the test if all six have disintegrated.
Effervescent tablets are uncoated tablets generally containing Dissolution (2.5.2). For tablets prepared from granules or
acidic substances and either carbonates or bicarbonates which particles already covered with an enteric coating, the
react rapidly in the presence of water to release carbon dioxide. dissolution test is carried out to demonstrate the appropriate
They are intended to be dissolved or dispersed in water before release of the active substance(s).
administration.
Tests Prolonged- release Tablets

Disintegration (2.5.1). Place one tablet in a 250-ml beaker Prolonged-release tablets, also known as sustained-release
containing water at 20º to 30º; numerous gas bubbles are tablets or extended-release tablets are tablets formulated in
evolved. When the evolution of gas around the tablet or its such a manner as to make the contained active ingredient
fragments has ceased the tablet shall have disintegrated, being available over an extended period of time after ingestion.
either dissolved or dispersed in the water so that no
agglomerates of particles remain. Repeat the operation on a Tests
further 5 tablets. The tablets comply with the test if each of
Dissolution (2.5.2). The test should be designed to
the 6 tablets disintegrates in the manner prescribed within 5
demonstrate the appropriate release of the active substance(s).
minutes, unless otherwise stated in the individual monograph.
The manufacturer is expected to give specifications for drug
release at 3 or more test-time points. The first point should be
Modified-release Tablets set after a testing period corresponding to a dissolved amount
Modified-release tablets (Sustained-release tablets) are coated of typically 20 per cent to 30 per cent. The second point should
or uncoated tablets containing auxiliary substances or define the dissolution pattern and should be set at around 50
prepared by procedures that, separately or together, are per cent release. The final point should ensure almost complete
designed to modify the rate or the place at which the active release that is generally understood as more than 80 per cent
ingredient is released. release.
Modified-release tablets include enteric-coated tablets, Carry out the test for the test-times indicated on the label of
prolonged-release tablets and delayed-release tablets. the product.
46
IP 2007 TABLETS
Soluble Tablets Tablets for Use in the Mouth

Soluble tablets are uncoated tablets or film-coated tablets that Tablets for use in the mouth are usually uncoated tablets
are to be dissolved in water before use. The solution produced formulated to be chewed or to effect a slow release and local
may be slightly opalescent due to added substances used in action of the active ingredient (lozenges) or the release and
the manufacture of the tablets. absorption of the active ingredient under the tongue
(sublingual tablets). Chewable tablets and lozenges may
Tests contain flavouring agents.
Disintegration (2.5.1). Soluble tablets disintegrate within 3
minutes. The test is carried out using water at 15º to 25º.
47
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS
DRUG SUBSTANCES, DOSAGE FORMS

AND
PHARMACEUTICAL AIDS
A to .......................................................................................................................
49
A
Abacavir Sulphate ....
Abacavir Oral Solution ....
Abacavir Tablets ....
Abacavir and Lamivudine Tablets ....
Abacavir, Lamivudin and Zidovudine Tablets ....
Acarbose ....
Acarbose Tablets ....
Acebutolol Hydrochloride ....
Acebutolol Tablets ....
Aceclofenac ....
Aceclofenac Tablets ....
Acetazolamide ....
Acetazolamide Tablets ....
Glacial Acetic Acid ....
Acetic Acid Ear Drops ....
Aciclovir ....
Aciclovir Intravenous Infusion ....
Aciclovir Tablets ....
Adenine ....
Adrenaline ....
Adrenaline Tartrate ....
Adrenaline Injection ....
Albendazole ....
Albendazole Tablets ....
Alginic Acid ....
Allopurinol ....
Allopurinol Tablets ....
Aloes ....
Alprazolam ....
Alprazolam Tablets ....
51
MONOGRAPHS INDIAN PHARMACOPOEIA 2007 2007
Aluminium Acetate Ear Drops ....

Aluminium Hydroxide Gel ....
Dried Aluminium Hydroxide Gel ....
Aluminium Sulphate ....
Amantadine Hydrochloride ....
Amantadine Capsules ....
Ambroxol Hydrochloride ....
Amikacin ....
Amikacin Sulphate ....
Amikacin Injection ....
Amiloride Hydrochloride ....
Amiloride Tablets ....
Aminocaproic Acid ....
Aminocaproic Acid Injection ....
Aminocaproic Acid Tablets ....
Aminophylline ....
Aminophylline Injection ....
Aminophylline Tablets ....
Amiodarone Hydrochloride ....
Amiodarone Tablets ....
Amitriptyline Hydrochloride ....
Amitriptyline Tablets ....
Amlodipine Besilate ....
Amlodipine Tablets ....
Ammonium Chloride ....
Amodiaquine Hydrochloride ....
Amodiaquine Tablets ....
Amoxycillin Sodium ....
Amoxycillin Capsules ....
Amoxycillin Injection ....
Amoxycillin Oral Suspension ....
Amoxycillin Trihydrate ....
52
Amoxycillin Dispersible Tablets ....

Amoxycillin and Potassium Clavulanate Injection ....
Amoxycillin and Potassium Clavulanate Oral Suspension ....
Amoxycillin and Potassium Clavulanate Tablets ....
Amphotericin B ....
Amphotericin B Injection ....
Ampicillin ....
Ampicillin Capsules ....
Ampicillin Sodium ....
Ampicillin Injection ....
Ampicillin Oral Suspension ....
Ampicillin Dispersible Tablet ....
Ampicillin Trihydrate ....
Alpha Amylase ....
Analgin ....
Analgin Tablets ....
Anticoagulant Citrate Dextrose Solution ....
Anticoagulant Citrate Phosphate Dextrose Solution ....
Anticoagulant Citrate Phosphate Dextrose Adenine Solution ....
Arteether ....
Artemether ....
Artemisinin ....
Ascorbic Acid ....
Ascorbic Acid Injection ....
Ascorbic Acid Tablets ....
Aspartame ....
Aspirin ....
Aspirin Tablets ....
Soluble Aspirin Tablets ....
Aspirin And Caffeine Tablets ....
Atenolol ....
Atenolol Tablets ....
Atorvastatin Calcium ....
53
Atorvastatin Tablets ....

Atropine Methonitrate ....
Atropine Sulphate ....
Atropine Injection ....
Atropine Eye Ointment ....
Atropine Tablets ....
Azathioprine ....
Azathioprine Tablets ....
Azithromycin ....
Azithromycin Capsules ....
Azithromycin Oral Suspension ....
Azithromycin Tablets ....
54
IP 2007 ABACAVIR ORAL SOLUTION
Heavy metals (2.3.13). 1.0 g complies with the limit test for
Abacavir Sulphate heavy metals, Method B (20 ppm).
Sulphated ash (2.3.18). Not more than 0.3 per cent.
Water (2.3.43). Not more than 1.5 per cent, determined on 0.2 g.
HN
Assay. Determine by liquid chromatography (2.4.14).
N N
Test solution. A 0.01 per cent w/v solution of the substance
N , H2SO4 under examination in the mobile phase.
H2N N
Reference solution. A 0.01 per cent w/v solution of abacavir
HO sulphate RS in the mobile phase.
2 Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with
(C14H18N6O)2,H2SO4 Mol. Wt. 670.8 octadecylsilane chemically bonded to porous silica or
Abacavir Sulphate is {(1S,4R)-4-[2-amino-6- ceramic microparticles (5 µm),
(cyclopropylamino)9H-purin-9-yl]cyclopent-2- – mobile phase: a mixture of 10 volumes of methanol, 15
enyl}methanol sulphate. volumes of acetonitrile and 75 volumes of a buffer
prepared by dissolving 1.15 g of ammonium dihydrogen
Abacavir Sulphate contains not less than 98.0 per cent and
phosphate and 2 g of tetrabutylammonium hydrogen
not more than 102.0 per cent of (C14H18N6O)2, H2SO4, calculated
sulphate in 1000 ml of water and adjusting the pH to 6.0
on the anhydrous basis.
with triethylamine,
Description. A white or almost white, crystalline powder. – flow rate. 1.2 ml per minute,
Identification – a 20 µl loop injector.
A. Determine by infrared absorption spectrophotometry (2.4.6). Inject the reference solution.The test is not valid unless the
Compare the spectrum with that obtained with abacavir column efficiency determined from the abacavir sulphate peak
sulphate RS or with the reference spectrum of abacavir is not less than 3000 theoretical plates, the tailing factor is not
sulphate. more than 2.0 and the relative standard deviation of replicate
B. In the Assay, the principal peak in the chromatogram injections is not more than 2.0 per cent.
obtained with the test solution corresponds to the peak in the Inject alternately the test solution and the reference solution.
chromatogram obtained with the reference solution. Calculate the percentage content of (C14H18N6O)2,H2SO4.
C. It gives reaction A of sulphates (2.3.1). Storage. Store at a temperature not exceeding 30°.
Tests
Specific optical rotation (2.4.22). –32.0° to –38.0°, determined Abacavir Oral Solution
in a 0.5 per cent w/v solution in methanol.
Abacavir Sulphate Oral Solution
Related substances. Determine by liquid chromatography
(2.4.14), as described in the Assay but using the following Abacavir Oral Solution contains a quantity of Abacavir
solutions. Sulphate equivalent to not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of abacavir
Test solution. A 0.05 per cent w/v solution of the substance C14H18N6O. It may contain one or more suitable buffers,
under examination in the mobile phase. colours, flavours, preservatives, stabilizers, sweeteners, and
Reference solution. Dilute 1 ml of the test solution to 200 ml suspending agents.
with the mobile phase.
Identification
Inject the test solution and the reference solution. Calculate
the content of each impurity in the chromatogram obtained In the Assay, the principal peak in the chromatogram obtained
with the test solution by comparing the peak area of each with the test solution corresponds to the peak in the
peak with the area of the principal peak in the chromatogram chromatogram obtained with the reference solution.
obtained with the reference solution. The content of any
Tests
individual impurity is not greater than 0.5 per cent and the
sum of all the impurities is not greater than 1.5 per cent. pH (2.4.24). 4.6 to 5.0.
55
ABACAVIR TABLETS IP 2007
Related substances. Determine by liquid chromatography – mobile phase: a mixture of 85 volumes of a buffer solution
(2.4.14). prepared by dissolving 1.15 g of ammonium dihydrogen
phosphate and 2 g of tetrabutyl ammonium hydrogen
NOTE — Prepare the solutions immediately before use.
Test solution. Weigh a quantity of the oral solution containing with triethylamine and filtering, and 15 volumes of
50 mg of abacavir, dissolve in 100 ml of the mobile phase and acetonitrile,
mix. – flow rate. 1.5 ml per minute,
Reference solution (a). A solution of abacavir sulphate RS – spectrophotometer set at 214 nm,
containing 0.05 per cent w/v of abacavir in the mobile phase. – a 20 µl loop injector.
Inject the reference solution. The test is not valid unless the
Reference solution (b). Dilute 1 ml of reference solution (a) to column efficiency is not less than 3000 theoretical plates, the
100 ml with the mobile phase. tailing factor is not more than 2.0 and the relative standard
Chromatographic system deviation for replicate injections is not more than 2.0 per cent.
– a stainless steel column 25 cm x 4.6 mm, packed with Inject the test solution and the reference solution.
octadecylsilane chemically bonded to porous silica (5
Determine the weight per ml of the oral solution (2.4.29) and
µm),
calculate the content of C14H18N6O weight in volume.
– mobile phase: a mixture of 85 volumes of a buffer
solution prepared by dissolving 1.15 g of ammonium Storage. Store at a temperature not exceeding 30°. Do not
dihydrogen phosphate and 2 g of tetrabutyl ammonium freeze.
hydrogen sulphate in 1000 ml of water, adjusting the Labelling. The label states the strength in terms of the
pH to 6.0 with triethylamine and filtering, and 15 volumes equivalent amount of abacavir.
of acetonitrile,
Abacavir Tablets
Inject reference solution (a). The test is not valid unless the
column efficiency is not less than 3000 theoretical plates and Abacavir Sulphate Tablets
the tailing factor is not more than 2.0. Abacavir Tablets contain not less than 90.0 per cent and not
Inject the test solution and reference solution (b). In the more than 110.0 per cent of the stated amount of abacavir,
chromatogram obtained with the test solution, the area of any C14H18N6O. The tablets may be coated.
secondary peak is not more than the area of the peak in the
chromatogram obtained with the reference solution (b) Identification
(1.0 per cent) and the sum of areas of all the secondary peaks In the Assay, the principal peak in the chromatogram obtained
is not more than twice the area of the peak in the chromatogram with the test solution corresponds to the peak in the
obtained with the reference solution (b) (2.0 per cent). chromatogram obtained with the reference solution.
Other tests. Complies with the tests stated under Oral liquids.
Tests
Dissolution (2.5.2).
Apparatus. No 1
Test solution. Weigh accurately a quantity of the oral solution Medium. 900 ml of 0.1 M hydrochloric acid.
containing 60 mg of abacavir, dissolve in 100.0 ml of the mobile Speed and time. 75 rpm and 15 minutes.
phase and mix. Dilute 5.0 ml of the solution to 50.0 ml with the
mobile phase. Withdraw a suitable volume of the medium and filter, discarding
the first few ml of the filtrate.
Reference solution. A 0.06 percent w/v solution of abacavir
sulphate RS in the mobile phase and filter. Dilute 5.0 ml of the Determine by liquid chromatography (2.4.14).
solution to 50.0 ml with the mobile phase. Test solution. Dilute the filtrate, if necessary, with the
Chromatographic system dissolution medium.
– a stainless steel column 15 cm x 4.6 mm, packed with Reference solution. A 0.075 per cent w/v solution of abacavir
octadecylsilane chemically bonded to porous silica (5 sulphate RS in the dissolution medium. Dilute 5 ml of the
µm), solution to 10 ml with the dissolution medium.
56
IP 2007 ABACAVIR AND LAMIVUDINE TABLETS
Chromatographic system Test solution. Weigh and powder 20 tablets. Weigh accurately
– a stainless steel column 15 cm x 4.6 mm, packed with a quantity of the powder containing 50 mg of abacavir, disperse
octadecylsilane bonded to porous silica (5 µm), in 100.0 ml of the mobile phase and filter. Dilute 5.0 ml of the
– column temperature 40º, filtrate to 50.0 ml with the mobile phase.
– mobile phase: a mixture of 85 volumes of a buffer solution Reference solution. A 0.060 per cent w/v solution of abacavir
prepared by dissolving 1.15 g of ammonium dihydrogen sulphate RS in the mobile phase. Dilute 5.0 ml of the solution
phosphate and 2 g of tetrabutyl ammonium hydrogen to 50.0 ml with the mobile phase.
sulphate in 1000 ml of distilled water and adjusting the
pH to 6.0 with triethylamine, and 15 volumes of Chromatographic system
acetonitrile. – a stainless steel column 15 cm x 4.6 mm, packed with
– flow rate. 1.5 ml per minute, octadecylsilane bonded to porous silica (5 µm),
– spectrophotometer set at 214 nm, – column temperature 40º,
– a 10 µl loop injector. – mobile phase: a mixture of 85 volumes of a buffer solution
prepared by dissolving 1.15 g of ammonium dihydrogen
Inject alternatively the test solution and the reference solution.
phosphate and 2 g of tetrabutyl ammonium hydrogen
D. Not less than 80 per cent of the stated amount of C14H18N6O. sulphate in 1000 ml of distilled water and adjusting the
Related substances. Determine by liquid chromatography pH to 6.0 with triethylamine, and 15 volumes of
(2.4.14). acetonitrile.
Test solution. Weigh and powder 20 tablets. Weigh accurately
a quantity of the powder containing 50 mg of abacavir, disperse
in 100 ml of the mobile phase and filter.
Reference solution (a). A solution of abacavir sulphate RS tailing factor is not more than 2.0, the column efficiency in not
containing 0.05 per cent w/v of abacavir in the mobile phase. less than 2000 theoretical plates and the relative standard
Reference solution (b). Dilute 1 ml of reference solution (a) to deviation for replicate injections is not more than 2.0 per cent.
100 ml with the mobile phase. Inject the test solution and the reference solution.
Chromatographic system
Calculate the content of C14H18N6O.
octadecylsilane bonded to porous silica (5 µm), Storage. Store protected from moisture at a temperature not
– mobile phase: a mixture of 85 volumes of a buffer solution exceeding 30º.
prepared by dissolving 1.15 g of ammonium dihydrogen Labelling. The label states the strength in terms of the
phosphate and 2 g of tetrabutyl ammonium hydrogen equivalent amount of abacavir.
with triethylamine, and 15 volumes of acetonitrile.
– flow rate. 1.2 ml per minute, Abacavir and Lamivudine Tablets
– a 20 µl loop injector. Abacavir Sulphate and Lamivudine Tablets
Inject reference solution (a). The test is not valid unless the Abacavir and Lamivudine Tablets contain not less than 90.0
column efficiency is not less than 3000 theoretical plates and per cent and not more than 110.0 per cent of the stated amounts
the tailing factor is not more than 2.0. of abacavir, C14H18N6O and lamivudine, C8H11N3O3S.
Inject the test solution and reference solution (b). In the
Identification
chromatogram obtained with the test solution, the area of any
secondary peak is not more than the area of the peak in the In the Assay, the principal peaks in the chromatogram obtained
chromatogram obtained with the reference solution (b) with the test solution correspond to the principal peaks in the
(1.0 per cent) and the sum of all the secondary peaks is not chromatogram obtained with the reference solution.
more than twice the area of the peak in the chromatogram
obtained with the reference solution (b) (2.0 per cent). Tests
Other tests. Comply with the tests stated under Tablets. Dissolution (2.5.2).
Water (2.3.43). Not more than 5.0 per cent, determined on Apparatus. No 1
0.5 g. Medium. 900 ml of 0.1 M hydrochloric acid.
Assay. Determine by liquid chromatography (2.4.14). Speed and time. 75 rpm and 30 minutes.
57
ABACAVIR, LAMIVUDINE AND ZIDOVUDINE TABLETS IP 2007
Withdraw a suitable volume of the medium and filter. Inject reference solution (a). The test is not valid unless the
Determine by liquid chromatography (2.4.14) column efficiency is not less than 3000 theoretical plates and
the tailing factor is not more than 2.0.
Test solution. The filtrate obtained as given above. Dilute the
filtrate if necessary, with the dissolution medium.
Reference solution. Dissolve 75 mg of abacavir sulphate RS secondary peak is not more than the area of the peak in the
and 30 mg of lamivudine RS in 10 ml of methanol and dilute to chromatogram obtained with the reference solution (b)
100 ml with the dissolution medium. (1.0 per cent) and the sum of all the secondary peaks is not
Use the chromatographic system described under Assay. more than twice the area of the peak in the chromatogram
obtained with the reference solution (b) (2.0 per cent).
Inject the reference solution. The relative standard deviation
Other tests. Comply with the tests stated under Tablets.
for replicate injections is not more than 2.0 per cent.
Water (2.3.43). Not more than 3.0 per cent, determined on
Inject the test solution and the reference solution.
0.5 g.
D. Not less than 80 per cent of the stated amounts of C14H18N6O
Assay. Determine by liquid chromatography (2.4.14)
and C8H11N3O3S.
Related substances. Determine by liquid chromatography a quantity of the powder containing 60 mg of abacavir, dissolve
(2.4.14). in 20 ml of 0.1 M hydrochloric acid and dilute to 100.0 ml with
Solvent mixture. 95 volumes of mobile phase A and 5 volumes methanol. Dilute 5.0 ml of the solution to 50.0 ml with the
of mobile phase B. mobile phase.
Test solution. Weigh accurately a quantity of the powdered Reference solution. Dissolve 35 mg of abacavir RS and 15 mg
tablets containing 100 mg of abacavir and disperse in 100 ml of lamivudine RS in 15 ml of 0.1 M hydrochloric acid and
of the solvent mixture and filter. dilute to 50.0 ml with methanol. Dilute 5.0 ml of the solution to
50.0 ml with the mobile phase.
Reference solution (a). A solution containing 0.12 per cent w/
v of abacavir sulphate RS and 0.05 per cent w/v of lamivudine Chromatographic system
RS in the solvent mixture. – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilane bonded to porous silica (5 µm),
Reference solution (b). Dilute 1 ml of reference solution (a) to – column temperature 40º,
100 ml with the solvent mixture. – mobile phase: a mixture of 50 volumes of a buffer solution
Chromatographic system prepared by dissolving 7.66 g of ammonium acetate in
– a stainless steel column 25 cm x 4.6 mm, packed with 1000 ml of a 0.5 per cent w/v solution of glacial acetic
octadecylsilane bonded to silica (5 µm), acid and 50 volumes of methanol,
– column temperature 40º, – flow rate. 1 ml per minute,
– mobile phase: A. a buffer solution prepared by – spectrophotometer set at 282 nm,
dissolving 1.9 g of ammonium acetate in 900 ml of water, – a 10 µl loop injector.
adjusting the pH to 3.8 with glacial acetic acid and Inject the reference solution. The test is not valid unless the
diluting to 1000 ml with water, relative standard deviation for replicate injections is not more
B. methanol, than 2.0 per cent.
– flow rate. 1 ml per minute,
– a linear gradient programme using the conditions given
below, Calculate the contents of C14H18N6O and C8H11N3O3S in the
– spectrophotometer set at 277 nm, tablets.
– a 20 µl loop injector. Storage. Store protected from moisture, at a temperature not
Time Mobile phase A Mobile phase B exceeding 30º.
(in min.) (per cent v/v) (per cent v/v)
0 95 5
20 95 5 Abacavir, Lamivudine and Zidovudine
40 30 70 Tablets
45 95 5 Abacavir, Lamivudine and Zidovudine Tablets contain not
50 95 5 less than 90.0 per cent and not more than 110.0 per cent of the
58
IP 2007 ABACAVIR, LAMIVUDINE AND ZIDOVUDINE TABLETS
stated amounts of abacavir, C14H18N6O lamivudine, C8H11N3O3S NOTE — Prepare the solutions immediately before use.
and zidovudine, C10H13N5O4. Solvent mixture. A 0.2 per cent v/v solution of
Identification orthophosphoric acid in a mixture of 70 volumes of water and
30 volumes of methanol.
In the Assay, the principal peaks in the chromatogram obtained
Test solution. Weigh a quantity of the powdered tablets
with the test solution correspond to the peaks in the
containing 75 mg of Lamivudine, disperse in 100 ml of the
chromatogram obtained with the reference solution.
solvent mixture and filter.
Tests Reference solution (a). A solution containing 0.18 per cent
w/v of abacavir sulphate RS, 0.075 per cent w/v lamivudine
RS and 0.15 per cent w/v of zidovudine RS in the solvent
Apparatus No. 1 mixture.
Medium. 900 ml of 0.1 M hydrochloric acid.
Reference solution (b). Dilute 1 ml of the solution to 100 ml
Speed and time. 75 rpm and 30 minutes. with the solvent mixture.
Withdraw a suitable volume of the medium and filter. Chromatographic system
Determine by liquid chromatography (2.4.14). – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilane bonded to porous silica (5µm),
Test solution. Use the filtrate obtained as given above.
– mobile phase: A. a mixture of 70 volumes of methanol,
Reference solution. A solution containing 0.035 per cent w/v 30 volumes of acetonitrileand 0.4 volume of
of abacavir sulphate RS, 0.015 per cent w/v lamivudine RS tetrahydrofuran,
and 0.03 per cent w/v of zidovudine RS in the dissolution B. a buffer solution pH 3.0 prepared by
medium. dissolving 6.8 g of potassium dihydrogen
Chromatographic system orthophosphate in 1000 ml of water, adjusting the pH
– a stainless steel column 5 cm × 4.6 mm, packed with to 3.0 with orthophosphoric acid and filtering,
octadecylsilane bonded to porous silica (3 µm) (such – a linear gradient programme using the conditions given
as Restek’s Pinnacle II C-18), below,
– column temperature 50º, – spectrophotometer set at 225 nm,
– mobile phase: a mixture of 88 volumes of a buffer solution – a 10 µl loop injector.
prepared by dissolving 1 g of octanesulphonic acid Time Mobile Mobile Flow rate
and 1 ml of triethylamine in 1000 ml of water and (min.) phase A phase B ml per minute
adjusting the pH to 2.5 with orthophosphoric acid, and 0 2 98 1
12 volumes of acetonitrile,
10 2 98 1
– flow rate. 2.5 ml per minute.
– spectrophotometer set at 272 nm, 25 20 80 1
– a 10 µl loop injector. 28 20 80 1
Inject the reference solution. The test is not valid unless the 50 30 70 1
resolution between lamivudine and zidovudine peaks is not 60 35 65 1.3
less than 2.5, the column efficiency determined from 63 35 65 1.3
lamivudine, zidovudine and abacavir peaks is not less than
66 2 98 1
700, 1200 and 2000 theoretical plates respectively, the tailing
factor for lamivudine, zidovudine and abacavir peaks is not 80 2 98 1
more than 2.0 and the relative standard deviation of replicate Inject reference solution (a). The test is not valid unless the
injections is not more than 2.0 per cent for each component. column efficiency is not less than 3000 and the tailing factor is
Inject the test solution and the reference solution. not more than 1.5 for each component.
Calculate the contents of C 14H 18N 6O, C 8H 11N 3O 3S and Inject the test solution and reference solution (b). In the
C10H13N5O4. chromatogram obtained with the test solution, the area of any
secondary peak is not more than 3 times the area of any peak
D. Not less than 70 per cent of the stated amounts of in the chromatogram obtained with the reference solution
C14H18N6O, C8H11N3O3S and C10H13N5O4. (3.0 per cent) and the sum of all the secondary peaks is not
Related substances. Determine by liquid chromatography more than 5 times the area of any peak due to the reference
(2.4.14). solution (5.0 per cent).
59
ACARBOSE IP 2007
Other tests. Comply with the tests stated under Tablets. glucopyranosyl-(1?4)-O-α-D- glucopyranosyl-(1→4)-D-
Assay. Determine by liquid chromatography (2.4.14). glucopyranose, which is produced by certain strains of
Actinoplanes utahensis.
Solvent mixture. 50 volumes of water and 50 volumes of
Acarbose contains not less than 95.0 per cent and not more
methanol.
than 102.0 per cent of C25H43NO18, calculated on the anhydrous
Test solution. Weigh accurately a quantity of the powdered basis.
tablets containing 150 mg of abacavir, dissolve in 100 ml of
Description. A white or yellowish, amorphous powder,
water, add 80 ml of methanol and dilute to 200.0 ml with
hygroscopic.
methanol. Dilute 10.0 ml of the solution to 25.0 ml with the
solvent mixture and filter.
Identification
Reference solution. A solution containing 0.35 per cent w/v
of abacavir sulphate RS, 0.15 per cent w/v lamivudine RS A. Determine by infrared absorption spectrophotometry (2.4.6).
and 0.30 per cent w/v of zidovudine RS in the solvent mixture. Compare the spectrum with that obtained with acarbose RS
Dilute 5.0 ml of the solution to 50.0 ml with the solvent mixture. or with the reference spectrum of acarbose.
Chromatographic system B. In the Assay, the principal peak in the chromatogram
– a stainless steel column 25 cm x 4.6 mm, packed with obtained with test solution corresponds to the peak in the
octadecylsilane bonded to porous silica (5µm), chromatogram obtained with the reference solution.
(such as Kromasil C-18),
– column temperature 50º, Tests
– mobile phase: a mixture of 65 volumes of a buffer solution pH (2.4.24). 5.5 to 7.5, determined in 5.0 per cent w/v solution
prepared by dissolving 1 g of octane sulphonic acid in carbon dioxide-free water (solution A).
and 1 ml of triethylamine in 1000 ml of water, adjusting
the pH to 4.5 with orthophosphoric acid and filtering, Specific optical rotation (2.4.22). +168º to +183º, dilute 2 ml of
and 35 volumes of methanol, solution A to 10 ml with water.
– flow rate. 1 ml per minute, Light absorption (2.4.7). Absorbance of solution A at 425 nm,
– spectrophotometer set at 272 nm, not more than 0.15.
Inject the reference solution. The test is not valid unless the (2.4.14).
column efficiency determined from the peak due to lamivudine
is not less than 2000 theoretical plates, the tailing factor is not Test solution. Dissolve 0.2 g of the substance under
more than 2.0 for each component and the relative standard examination in 100 ml of water.
deviation of replicate injections is not more than 2.0 per cent Reference solution (a). A 0.2 per cent w/v solution of
for each component. acarbose RS in water.
Inject the test solution and the reference solution. Reference solution (b). Dilute 1 ml of reference solution (a) to
Calculate the contents of (C14H18N6O)2,H2SO4, C8H11N3O3S and 100 ml with water.
C10H13N5O4 in the tablets. Chromatographic system as described under Assay.
Storage. Store protected from moisture, at a temperature not Inject reference solution (a). Test is not valid unless the column
exceeding 30º. effciency is not less than 2000 theoretical plates and the tailing
factor is not more than 2.0.
Acarbose chromatogram obtained with the test solution, the area of any
secondary peak is not more than 0.5 times the area of the peak
HO HO HO
CH3 in the chromatogram obtained with reference solution (b)
O O O
OH
(0.5 per cent) and the sum of areas of all the secondary peaks
OH OH OH OH
OH N O O is not more than twice the area of the peak in the chromatogram
H
OH OH OH OH obtained with the reference solution (b) (2.0 per cent).
C25H43NO18 Mol. Wt. 646.0 Heavy metals (2.3.13). 1 g complies with limit test for heavy
metals, Method B (20 ppm).
Acarbose is O-4,6-dideoxy-4-[[(1S,4R,5R,5S,6S)-4,5,6-
trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino-α-D- Sulphated ash (2.3.18). Not more than 0.2 per cent.
60
IP 2007 ACEBUTOLOL HYDROCHLORIDE
Water (2.3.43). Not more than 4.0 per cent, determined on Test solution. The filtrate diluted to produce a 0.002 per cent
0.3 g. w/v solution.
Assay. Determine by liquid chromatography (2.4.14). Reference solution. A 0.002 per cent w/v solution of
acarbose RS in solvent mixture.
Test solution. Dissolve 10 mg of the substance under
examination in 50.0 ml of water. Dilute 5.0 ml of the solution to Chromatographic system as described under Assay.
50.0 ml with water.
Calculate the content of C25H43NO18.
Reference solution. A 0.002 per cent w/v solution of D. Not less than 70 per cent of the stated amount of
acarbose RS in water. C25H43NO18.
– a stainless steel column 25 cm x 4 mm packed with
aminopropylsilyl silica (5 µm), Assay. Determine by liquid chromatography (2.4.14).
– mobile phase: a mixture of 75 volumes of acetonitrile Solvent mixture. 15 volumes of phsophate buffer pH 3.0 and
and 25 volumes of a solution containing 0.06 per cent 85 volumes of acetonitrile.
w/v of potassium dihydrogen phosphate and 0.035 per
cent w/v of disodium hydrogen phosphate dihydrate, Test solution. Weigh accurately a quantity containing 20 mg
– flow rate. 2 ml per minute, of Acarbose, disperse in 100.0 ml of solvent mixture.
– spectrophotometer set at 210 nm, Reference solution. A 0.02 per cent w/v solution of
– a 10 µl loop injector. acarbose RS in solvent mixture.
Inject the reference solution. The test is not valid unless the Chromatographic system
relative standard deviation for replicate injections is not more – a stainless steel column 25 cm x 4.6 mm, packed with
than 2.0 per cent. porous silica particles ( 5 µm),
Inject the test solution and the reference solution. – column temperature 40°,
– mobile phase: a mixture of 30 volumes of phosphate
Calculate the content of C25H43NO18. buffer pH 3.0 and 70 volumes of acetonitrile ,
Storage. Store protected from moisture. – flow rate. 1.2 ml per minute,
Acarbose Tablets relative standard deviation is not less than 2.0 per cent.
Acarbose Tablets contain not less than 90.0 per cent and not Inject the test solution and the reference solution.
more than 110.0 per cent of the stated amount of acarbose,
C25H43NO18.
Storage. Store protected from light and moisture.
Identification
In the Assay, the chromatogram obtained with the test solution
corresponds to the chromatogram obtained with the reference Acebutolol Hydrochloride
solution.
Tests CH3
O
OH H
O N CH3 , HCl
Apparatus No. 1 O
Medium. 900 ml of water. CH3
H 3C N
Speed and time. 100 rpm for 30 minutes. H
Withdraw a suitable volume of the medium and filter.
Determine by liquid chromatography (2.4.14). C18H28N2O4,HCl Mol. Wt. 372.9
Solvent mixture. 15 volumes of phsophate buffer pH 3.0 and Acebutolol Hydrochloride is (RS)-3'-acetyl-4'-(2-hydroxy-3-
85 volumes of acetonitrile. isopropylaminopropoxy)butyranilide hydrochloride.
61
ACEBUTOLOL TABLETS IP 2007
Acebutolol Hydrochloride contains not less than 99.0 per cent Mobile phase (b). A mixture of 90 volumes of 2-propanol and
and not more than 101.0 per cent of C18H28N2O4, HCl, calculated 10 volumes of glacial acetic acid.
on the dried basis. Test solution. Shake 0.5 g of the substance under examination
Description. A white or almost white, crystalline powder. with 30 ml of methanol in a 50-ml volumetric flask for 15 minutes,
dilute to volume with methanol.
Identification
Reference solution (a). Dilute 1 ml of the test solution to 10 ml
Test A may be omitted if tests B, C and D are carried out. Tests
with methanol.
B, C and D may be omitted if test A is carried out.
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution (b). Dilute 3 ml of reference solution (a) to
Compare the spectrum with that obtained with acebutolol 100 ml with methanol.
hydrochloride RS or with the reference spectrum of acebutolol Reference solution (c). A 0.1 per cent w/v solution of
hydrochloride. acebutolol hydrochloride RS in methanol.
B. When examined in the range 220 nm to 360 nm (2.4.7), a Reference solution (d). Dilute 1 volume of reference solution
0.001 per cent w/v solution in 0.1 per cent v/v solution of (c) to 3 volumes with methanol.
hydrochloric acid shows absorption maxima at about 233 nm
and 322 nm; absorbance at 233 nm, 0.55 to 0.61. Apply 20 µl of each solution on each plate. Develop two
chromatograms using separately the two mobile phases. After
C. Determine by thin layer chromatography (2.4.17), coating development, dry the plates in a current of warm air and
the plate with silica gel GF254. examine in ultraviolet light at 254 nm. Any secondary spot in
Mobile phase. A mixture of 60 volumes of water, 40 volumes the chromatograms obtained with the test solution is not more
of methanol and 0.5 volumes of perchloric acid. intense than the spot in the chromatogram obtained with
Test solution. Dissolve 0.1 g of the substance under reference solution (b) and not more than two such spots are
examination in 100 ml of methanol. more intense than the spot in the chromatograms obtained
with reference solution (d). Ignore any spot at the point of
Reference solution (a). A 0.1 per cent w/v solution of application.
acebutolol hydrochloride RS in methanol.
Assay. Weigh and powder 20 tablets. Weigh accurately a
Reference solution (b). A mixture of equal volumes of reference quantity of the powder containing about 0.1 g of Acebutolol
solution (a) and a 0.1 per cent w/v solution of pindolol RS in Hydrochloride, shake with 40 ml of 0.1 M hydrochloric acid
methanol. and add sufficient water to produce 100.0 ml, filter and dilute
Apply to the plate 10 µl of each solution. After development, 10.0 ml of the filtrate to 100.0 ml with water. Dilute 10.0 ml of
dry the plate in a current of warm air and examine in ultraviolet this solution to 100.0 ml with water and measure the absorbance
light at 254 nm. The principal spot in the chromatogram of the resulting solution at the maximum at about 233 nm (2.4.7).
obtained with the test solution corresponds to that in the Calculate the content of C18H28N2O4,HCl taking 580 as the
chromatogram obtained with reference solution (a). The test specific absorbance at 233 nm.
is not valid unless the chromatogram obtained with reference
solution (b) shows two clearly separated spots. Storage. Store protected from light.
D. A 5 per cent w/v solution gives reaction A of chlorides
(2.3.1).
Acebutolol Tablets
Tests
Acebutolol HydrochlorideTablets
Appearance of solution. A 5.0 per cent w/v solution is not
Acebutolol Tablets contain Acebutolol Hydrochloride. The
more opalescent than opalescence standard OS2 (2.4.1) and
tablets may be coated.
not more intensely coloured than reference solution BYS5
(2.4.1). Acebutolol Tablets contain not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of acebutolol
pH (2.4.24). 5.0 to 7.0, determined in a 1.0 per cent w/v solution.
hydrochloride, C18H28N2O4, HCl.
Related substances. Determine by thin layer chromatography
(2.4.17), coating two plates with silica gel GF 254. Identification
Mobile phase (a). The upper layer obtained by shaking A. When examined in the range 220 nm to 360 nm (2.4.7), the
together 50 volumes of water, 40 volumes of 1-butanol and 10 solution obtained in the Assay, shows an absorption maximum
volumes of glacial acetic acid. at about 233 nm.
62
IP 2007 ACECLOFENAC
B. Determine by thin layer chromatography (2.4.17), coating the chromatograms obtained with the test solution is not more
the plate with silica gel GF254. intense than the spot in the chromatogram obtained with
Mobile phase. A mixture of 60 volumes of water, 40 volumes reference solution (b) and not more than two such spots are
of methanol and 0.5 volumes of perchloric acid. more intense than the spot in the chromatograms obtained
with reference solution (d). Ignore any spot at the point of
Test solution. Shake a quantity of the powdered tablets application.
containing 0.5 g of Acebutolol Hydrochloride with 30 ml of
methanol in a 50-ml volumetric flask for 15 minutes, dilute to
volume with methanol, centrifuge and use the clear Assay. Weigh and powder 20 tablets. Weigh accurately a
supernatant liquid. quantity of the powder containing about 0.1 g of Acebutolol
Hydrochloride, shake with 40 ml of 0.1M hydrochloric acid
Reference solution (a). A 0.1 per cent w/v solution of
and add sufficient water to produce 100.0 ml, filter and dilute
acebutolol hydrochloride RS in methanol.
10.0 ml of the filtrate to 100.0 ml with water. Dilute 10.0 ml of
Reference solution (b). A mixture of equal volumes of reference this solution to 100.0 ml with water and measure the absorbance
solution (a) and a 0.1 per cent w/v solution of pindolol RS in of the resulting solution at the maximum at about 233 nm (2.4.7).
methanol.
Calculate the content of C18 H28 N2O4,HCl taking 580 as the
Apply to the plate 10 µl of each solution. After development, specific absorbance at 233 nm.
dry the plate in a current of warm air and examine in ultraviolet
Storage. Store protected from light.
light at 254 nm. The principal spot in the chromatogram
obtained with the test solution corresponds to that in the
chromatogram obtained with reference solution (a). The test
is not valid unless the chromatogram obtained with reference Aceclofenac
solution (b) shows two clearly separated spots.
O COOH
Tests
O
Related substances. Determine by thin layer chromatography NH
(2.4.17), coating two plates with silica gel GF 254.
Cl Cl
Mobile phase (a). The upper layer obtained by shaking
together 50 volumes of water, 40 volumes of 1-butanol and 10
volumes of glacial acetic acid.
C16H13Cl2NO4 Mol. Wt. 354.2
Mobile phase (b). A mixture of 90 volumes of 2-propanol and
10 volumes of glacial acetic acid. Aceclofenac is [(2,6-dichlorophenyl)amino]
phenylacetoxyacetic acid.
Test solution. Shake a quantity of the powdered tablets
containing 0.5 g of Acebutolol Hydrochloride with 30 ml of Aceclofenac contains not less than 99.0 per cent and not
methanol in a 50-ml volumetric flask for 15 minutes, dilute to more than 101.0 per cent of C16H13Cl2NO4, calculated on the
volume with methanol, centrifuge and use the clear dried basis.
supernatant liquid. Description. A white or almost white, crystalline powder.
with methanol. Identification
Reference solution (b). Dilute 3 ml of reference solution (a) to Test A may be omitted if tests B and C are carried out. Tests B
100 ml with methanol. and C may be omitted if test A is carried out.
Reference solution (c). A 0.1 per cent w/v solution of A. Determine by infrared absorption spectrophotometry (2.4.6).
acebutolol hydrochloride RS in methanol. Compare the spectrum with that obtained with aceclofenac
RS or with the reference spectrum of aceclofenac.
Reference solution (d). Dilute 1 volume of reference solution
(c) to 3 volumes with methanol. B. When examined in the range 220 nm to 370 nm (2.4.7), the
0.002 per cent w/v solution in methanol shows an absorption
Apply 20 µl of each solution on each plate. Develop two
maximum at 275 nm .
chromatograms using separately the two mobile phases. After
development, dry the plates in a current of warm air and C. Dissolve about 10 mg in 10 ml of ethanol. To 1 ml of the
examine in ultraviolet light at 254 nm. Any secondary spot in solution, add 0.2 ml of a mixture, prepared immediately before
63
ACECLOFENAC TABLETS IP 2007
use, of equal volumes of a 0.6 per cent solution of potassium Heavy metals (2.3.13). 2.0 g complies with limit test for heavy
ferricyanide and a 0.9 per cent solution of ferric chloride. metals, Method B ( 10 ppm ).
Allow to stand protected from light for 5 minutes. Add 3 ml of Sulphated ash (2.3.18). Not more than 0.1 per cent.
a 1 per cent solution of hydrochloric acid. Allow to stand
protected from light for 15 minutes. A blue colour develops Loss on drying (2.4.19). Not more than 0.5 per cent, determined
and a precipitate is formed. on 1 g by drying in an oven at 105°.
Assay. Weigh accurately about 0.3 g and dissolve in 40 ml of
Tests methanol. Titrate with 0.1 M sodium hydroxide. Determine
Related substances. Determine by liquid chromatography the end point potentiometrically (2.4.25). Carry out a blank
(2.4.14). titration.
Solvent mixture. A mixture of 30 volumes of mobile phase A 1 ml of 0.1 M sodium hydroxide is equivalent to 0.03542 g of
and 70 volumes of mobile phase B. C16H13Cl2NO4.
examination in 25 ml in solvent mixture.
aceclofenac RS in solvent mixture.
Aceclofenac Tablets
Reference solution (b). Dilute 1 ml of reference solution (a) to
100 ml with solvent mixture. Aceclofenac Tablets contain not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of
Chromatographic system aceclofenac, C16H13Cl2NO4.
– a stainless steel column 25 cm x 4.6 mm packed with
spherical end-capped octadecylsilane bonded to Identification
porous silica (5 µm), with a pore size of 10 µm and carbon
loading of 19 per cent, In the Assay, the principal peak in the chromatogram obtained
– mobile phase: A. a 0.112 per cent w/v solution of with the test solution corresponds to the peak in the
orthophosphoric acid adjusted to pH 7.0 using a chromatogram obtained with the reference solution.
4.2 per cent solution of sodium hydroxide,
B. 1 volume of water and 9 volumes of Tests
acetonitrile, Dissoluton (2.5.2).
below, Apparatus. No. 1
– flow rate. 1 ml per minute, Medium: 900 ml of phosphate buffer pH 7.5.
– spectrophotometer set at 275 nm, Speed and time. 50 rpm for 45 minutes.
– a 10 µl loop injector. Withdraw a suitable volume of the medium and filter. Reject
Time Mobile phase A Mobile phase B the first few ml of the filtrate and dilute a suitable volume of
(in min) (per cent v/v) (per cent v/v) the filtrate with dissolution medium. Measure the absorbance
0 70 30 of the resulting solution at the maximum at about 273 nm (2.4.7).
25 50 50 Calculate the content of aceclofenac, C16H13Cl2NO4 in the
medium from the absorbance obtained from a solution of
30 20 80 known concentration of aceclofenac RS.
50 20 80
D. Not less than 70 per cent of the stated amount of
52 70 30 C16H13Cl2NO4.
65 70 30
Inject the test solution and reference solution (b). In the (2.4.14).
Test solution. Weigh accurately a quantity of powdered tablet
containing 100 mg of Aceclofenac, disperse in 100 ml of mobile
in the chromatogram obtained with reference solution (b) (0.5
phase and filter.
per cent) and the sum of areas of all the secondary peaks is
not more than twice the area of the peak in the chromatogram Reference solution (a). A 0.1 per cent w/v solution of
obtained with the reference solution (b) (2.0 per cent). aceclofenac RS in the mobile phase.
64
IP 2007 ACETAZOLAMIDE
Reference solution (b). Dilute 1 ml of reference solution (a) to tailing factor is not more than 2.0.The relative standard
100 ml with mobile phase. deviation for replicate injections is not more than 2.0 per cent.
Chromatographic system Inject the test solution and the reference solution.
– a stainless steel column 25 cm x 4.6 mm packed with Calculate the content of C16H13Cl2NO4.
dimethyloctylsilane (C8 alkyl chain) fully endcapped
stationary phase (5 µm) (such as Hypersil MOS), Storage. Store protected from moisture, at a temperature not
– mobile phase: a mixture of 55 volumes of buffer pH 3.5 exceeding 25°.
prepared by adding 1.2 ml of glacial acetic acid in 1000
ml of water. adjust the pH to 3.5 with dilute sodium
hydroxide and filter, 22.5 volumes of acetonitrile and Acetazolamide
22.5 volumes of tetrahydrofuran,
– flow rate. 1 ml per minute, O H
O S S N
– spectrophotometer set at 275 nm, CH3
– a 20 µl loop injector. H 2N N N O
Inject reference solution (a). Test is not valid unless the column
effciency is not less than 2000 theoretical plates and the tailing C4H6N4O3S2 Mol. Wt. 222.2
factor is not more than 2.0. Acetazolamide is N-(5-sulphamoyl-1,3,4-thiadiazol-2-yl)
Inject the test solution and reference solution (b). In the acetamide.
chromatogram obtained with the test solution, the area of any Acetazolamide contains not less than 98.5 per cent and not
secondary peak is not more than the area of the peak in the more than 101.0 per cent of C4H6N4O3S2, calculated on the
chromatogram obtained with reference solution (b) (1.0 per dried basis.
cent) and the sum of areas of all the secondary peaks is not
more than twice the area of the peak in the chromatogram Description. A white to faintly yellowish-white, crystalline
obtained with the reference solution (b) (2.0 per cent). powder; odourless.
Other tests. Comply with the tests stated under Tablets. Identification
Solvent mixture. A mixture of 55 volumes of acetonitrile and C and D may be omitted if tests A and B are carried out.
45 volumes of water. A. Determine by infrared absorption spectrophotometry (2.4.6).
Test solution. Weigh and powder 20 tablets. Weigh accurately Compare the spectrum with that obtained with acetazolamide
a quantity of powdered tablet containing 100 mg of RS or with the reference spectrum of acetazolamide.
Aceclofenac, add about 60 ml of acetonitrile and sonicate for B. When examined in the range 230 nm to 260 nm (2.4.7), a
10 minutes. Make up the volume to 100.0 ml with acetonitrile. 0.003 per cent w/v solution in 0.01 M sodium hydroxide shows
Dilute 5.0 ml of the solution to 50.0 ml with solvent mixture. an absorption maximum at about 240 nm; absorbance at about
Reference solution. Weigh about 25 mg of aceclofenac RS 240 nm, 0.49 to 0.53. When examined in the range 260 nm to
and add acetonitrile to dissolve and make up the volume to 360 nm (2.4.7), a 0.00075 per cent w/v solution in 0.01 M sodium
25.0 ml with solvent mixture. Dilute 5.0 ml of the above solution hydroxide shows an absorption maximum at about 292 nm;
to 50.0 ml with solvent mixture. absorbance at about 292 nm, 0.43 to 0.47.
Chromatographic system C. To about 20 mg in a test-tube add 4 ml of 2 M hydrochloric
– a stainless steel column 15 cm x 4.6 mm packed with acid and 0.2 g of zinc powder and immediately place a piece of
octadecylsilane stationary phase (5 µm) (such as lead acetate paper over the mouth of the tube; the paper
Hypersil ODS), exhibits a brownish-black colour.
– mobile phase: a mixture of 55 volumes of buffer solution D. To about 25 mg add 5 ml of water, 4 drops of 1 M sodium
prepared by adding 1.0 ml of glacial acetic acid in hydroxide and 2 drops of cupric sulphate solution; a bluish-
1000 ml of water and 45 volumes of acetonitrile, green colour or precipitate is produced.
– spectrophotometer set at 275 nm, Tests
Silver-reducing substances. Mix 5 g with 25 ml of ethanol
Inject the reference solution. The test is not valid unless the (95 per cent), add 125 ml of water, 10 ml of nitric acid and 5 ml
column efficiency is not less than 2500 theoretical plates. The of 0.1 M silver nitrate, stir for 30 minutes and filter. Wash the
65
ACETAZOLAMIDE TABLETS IP 2007
residue with water, mix the filtrate and washings and titrate thoroughly and filter. Neutralise the filtrate with glacial acetic
the excess of silver nitrate in the mixture with 0.05 M acid, filter and dry the resulting precipitate at 105°. The residue
ammonium thiocyanate using ferric ammonium sulphate complies with the following test.
solution as indicator; not less than 9.5 ml of 0.05 M ammonium
Determine by infrared absorption spectrophotometry (2.4.6).
thiocyanate is required.
Compare the spectrum with that obtained with acetazolamide
Related substances. Determine by thin-layer chromatography RS or with the reference spectrum of acetazolamide.
(2.4.17), coating the plate with silica gel GF254.
B. Triturate a quantity of the powdered tablets containing 0.5
Mobile phase. A freshly prepared mixture of 50 volumes of 2- g of Acetazolamide with a mixture of 5 ml of water and 1 ml of
propanol, 30 volumes of ethyl acetate and 20 volumes of 1 M sodium hydroxide, transfer to a test-tube, add 0.2 g of
strong ammonia solution. zinc powder and immediately place a piece of lead acetate
Test solution. Dissolve 0.5 g of the substance under paper over the mouth of the tube; the paper exhibits a
examination in 100 ml of a mixture of equal volumes of ethanol brownish-black colour.
(95 per cent) and ethyl acetate.
C. To a quantity of the powdered tablets containing 25 mg of
Reference solution. A 0.005 per cent w/v solution of the Acetazolamide add 5 ml of water, 3 drops of 1 M sodium
substance under examination in a mixture of equal volumes of hydroxide and 2 drops of cupric sulphate solution; a bluish-
ethanol (95 per cent) and ethyl acetate. green colour or precipitate is produced.
Apply to the plate 20 µl of each solution. Do not line the walls
of the tank. Allow to saturate for 1 hour before development. Tests
After development, dry the plate in a current of warm air and
Related substances. Determine by thin-layer chromatography
examine in ultraviolet light at 254 nm. Any secondary spot in
the chromatogram obtained with the test solution is not more
intense than the spot in the chromatogram obtained with the Mobile phase. A freshly prepared mixture of 50 volumes of 2-
reference solution. propanol, 30 volumes of ethyl acetate and 20 volumes of
Heavy metals (2.3.13). 1.0 g dissolved in a mixture of 10 ml of 1 strong ammonia solution.
M sodium hydroxide and 15 ml of water complies with the Solvent mixture. A mixture of equal volumes of ethanol (95
limit test for heavy metals, Method C (20 ppm). per cent) and ethyl acetate.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Test solution. Shake a quantity of the powdered tablets
Loss on drying (2.4.19). Not more than 0.5 per cent, determined containing 50 mg of Acetazolamide for 20 minutes with 10 ml
on 2.5 g by drying in an oven at 105°. of solvent mixture, filter and use the filtrate.
Assay. Weigh accurately about 0.4 g of the substance under Reference solution. Dilute 1 volume of the test solution to 100
examination and dissolve in 90 ml of dimethylformamide. volumes with the same solvent mixture.
Titrate with 0.1 M tetrabutylammonium hydroxide,
Apply to the plate 20 µl of each solution. Do not line the walls
determining the end-point potentiometrically (2.4.25). Take
of the tank. Allow to saturate for 1 hour before development.
precautions to prevent absorption of atmospheric carbon
After development, dry the plate in a current of warm air and
dioxide. Carry out a blank titration.
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to the chromatogram obtained with the test solution is not more
0.02222 g of C4H6 N4O3S2. intense than the spot in the chromatogram obtained with the
Storage. Store protected from light. reference solution.
Acetazolamide Tablets
quantity of the powder containing about 0.4 g of
Acetazolamide Tablets contain not less than 95.0 per cent and Acetazolamide and add 90 ml of dimethylformamide. Titrate
not more than 105.0 per cent of the stated amount of with 0.1 M tetrabutylammonium hydroxide, determining the
acetazolamide, C4H6N4O3S2. end-point potentiometrically (2.4.25). Carry out a blank titration.
Identification 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
0.02222 g of C4H6N4O3S2.
A. To a quantity of the powdered tablets containing 0.5 g of
Acetazolamide add 2 ml of 1 M sodium hydroxide, shake Storage. Store protected from light.
66
IP 2007 ACICLOVIR
Glacial Acetic Acid 1 ml of 1 M sodium hydroxide is equivalent to 0.06005 g of

C2H4O2.
O Storage. Store protected from light and moisture.
H3C OH
C2H4O2 Mol. Wt. 60.1 Acetic Acid Ear Drops
Glacial Acetic acid contains not less than 99.0 per cent w/w Acetic Acid Otic Solution
and not more than 100.5 per cent w/w of C2H4O2.
Acetic Acid Ear Drops is a solution of Glacial Acetic Acid in a
Description. A crystalline mass or clear, colourless, volatile suitable non-aqueous solvent.
liquid.
Acetic Acid Ear Drops contain not less than 85.0 per cent and
Identification not more than 130.0 per cent of the stated amount of acetic
acid, C2H4O2.
A. A 10 per cent w/v solution is strongly acidic.
B. To 0.03 ml add 3 ml of water and neutralize with 2 M sodium Identification
hydroxide; the solution gives reaction C of acetates (2.3.1).
A. Dilute 5 ml with 10 ml of water and adjust to a pH of about
Tests 7 with 1 M sodium hydroxide. Add ferric chloride test solution,
a deep red colour is produced, which is decolorized on the
Freezing point (2.4.11). Not less than 14.8°. addition of hydrochloric acid.
Residue on evaporation. Not more than 0.01 per cent, determined
B. Warm the solution with sulphuric acid and ethanol (95
on 20.0 g by evaporating to dryness on a water-bath and
per cent); a characteristic odour of ethyl acetate is evolved.
drying at 105°.
Reducing substances. To 5 ml add 10 ml of water and mix. To Tests
5 ml of the resulting solution add 6 ml of sulphuric acid and
cool. Add 2 ml of 0.0167 M potassium dichromate, allow to pH (2.4.24). 2.0 to 4.0, determined in a 50.0 per cent v/v solution.
stand for 1 minute and add 25 ml of water and 1 ml of freshly Other tests. Comply with the tests stated under Ear Drops.
prepared dilute potassium iodide solution.Titrate with 0.1 M
Assay. Transfer a volume containing about 0.1 g of Glacial
sodium thiosulphate using 1 ml of starch solution as indicator.
Acetic Acid to a conical flask, add 5 ml of sodium chloride
Not less than 1.0 ml of 0.1 M sodium thisulphate is required.
solution and about 40 ml of water. Titrate with 0.1 M sodium
Heavy metals (2.3.13). Dissolve the residue obtained in the hydroxide, using 0.15 ml of phenolphthalein solution as
test for Residue on evaporation by heating with two quantities, indicator.
each of 15 ml, of water and add sufficient water to produce 50
ml (solution A). The solution complies with the limit test for 1 ml of 0.1 M sodium hydroxide is equivalent to 0.006005 g of
heavy metals, Method D (5 ppm). Use 10 ml of lead standard C2H4O2.
solution (2 ppm Pb) to prepare the standard. Storage. Store protected from light and moisture.
Iron (2.3.14). 5 ml of solution A diluted to 10 ml with water
complies with the limit test for iron (5 ppm). Use 1.0 ml of iron
standard solution (10 ppm Fe) to prepare the standard. Aciclovir
Chlorides (2.3.12). To 20 ml add sufficient water to produce Acyclovir
100 ml (solution B). 10 ml of solution B diluted to 15 ml with
water complies with the limit test for chlorides (25 ppm). Use O
10 ml of chloride standard solution (5 ppm Cl) to prepare the
HN N
standard.
Sulphates (2.3.17). 15 ml of solution B complies with the limit N
H 2N N
test for sulphates (50 ppm).
OH
Assay. Weigh accurately a conical flask with a ground-glass O
stopper containing 25 ml of water, add 1 ml of the substance
C8H11N5O3 Mol. Wt. 225.2
under examination and reweigh. Titrate with 1 M sodium
hydroxide using 0.5 ml of phenolphthalein solution of Aciclovir is 2-amino-9-[2-hydroxyethoxy)methyl]-1,9-dihydro-
indicator. 6H-purin-6-one.
67
ACICLOVIR INTRAVENOUS INFUSION IP 2007
Aciclovir contains not less than 98.5 per cent and not more suitable alkali, with or without auxiliary substances. It is filled
than 101.0 per cent of C8H11N5O3, calculated on the anhydrous in a sealed container.
basis. The infusion is constituted by dissolving the contents of the
Description. A white or almost white, crystalline powder. sealed container in the requisite amount of sterile Water for
Injections, immediately before use.
Identification
The constituted solution complies with the requirements for
Determine by infrared absorption spectrophotometry (2.4.6). Clarity of solution and Particulate matter stated under
Compare the spectrum with that obtained with aciclovir RS. Parenteral Preparations (Injections).
Tests Storage. The constituted solution should be used immediately
after preparation but, in any case, within the period
Appearance of solution. A 1.0 per cent w/v solution in 0.1 M
recommended by the manufacturer.
sodium hydroxide is clear (2.4.1), and not more intensely
coloured than reference solution YS7 (2.4.1). Aciclovir Intravenous Infusion contains not less than 95.0
Related substances. Determine by thin-layer chromatography per cent and not more than 105.0 per cent of the stated amount
(2.4.17), coating the plate with silica gel GF254. of aciclovir, C8H11N5O3.
Mobile phase. A mixture of 80 volumes of dichloromethane, Description. A white or almost white, crystalline powder.
20 volumes of methanol and 2 volumes of strong ammonia The contents of the sealed container comply with the
solution. requirements stated under Parenteral Preparations
Prepare the following solutions immediately before use. (Powders for Injections) and with the following requirements.
Test solution. Dissolve 1.0 g of the substance under Identification
examination in 100 ml of dimethyl sulphoxide.
A. When examined in the range 230 nm to 360 nm (2.4.7), the
Reference solution. A 0.005 per cent w/v solution of aciclovir solution prepared in the Assay shows an absorption maximum
impurity RS in dimethyl sulphoxide. at about 255 nm and a broad shoulder at about 274 nm.
Apply to the plate 10 µl of each solution. Keep the spots B. In the test for Guanine, the principal spot in the
compact by drying in a current of warm air and allow the plate chromatogram obtained with test solution (b) corresponds to
to cool. Allow the mobile phase to rise 10 cm. Dry the plate in that in the chromatogram obtained with reference solution (a).
air and examine in ultraviolet light at 254 nm. Any secondary
spot with Rf value greater than that of the principal spot in the C. Gives reaction A of sodium salts (2.3.1).
chromatogram obtained with the test solution is not more
intense than the spot in the chromatogram obtained with the Tests
reference solution (0.5 per cent). Appearance of solution. Dissolve the contents of a sealed
Sulphated ash (2.3.18). Not more than 0.1 per cent. container in sufficient water for injection to produce a
solution containing the equivalent of 2.5 per cent w/v solution
of Aciclovir (solution A). The solution is not more opalescent
0.5 g.
than opalescence standard OS2 (2.4.1), and not more intensely
Assay. Weigh accurately about 0.15 g and dissolve in 60 ml of coloured than reference solution BYS5 (2.4.1).
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
pH (2.4.24). 10.7 to 11.7, determined in solution A.
acid, determining the end-point potentiometrically (2.4.25).
Carry out a blank titration. Guanine. Determine by thin-layer chromatography (2.4.17),
1 ml of 0.1 M perchloric acid is equivalent to 0.02252 g of coating the plate with cellulose F254. (Merck celllulose F
C8H11N5O3. plates are suitable).
Storage. Store protected from light and moisture. Mobile phase. A mixture of 10 volumes of 1-propanol, 30
volumes of strong ammonia solution and 60 volumes of a 5
per cent w/v solution of ammonium sulphate.
Aciclovir Intravenous Infusion Test solution (a). Dissolve a suitable quantity of the substance
Acyclovir Intravenous Infusion; Acyclovir Sodium under examination in sufficient 0.1 M sodium hydroxide to
Intravenous Infusion produce a solution containing 0.5 per cent of Aciclovir.
Aciclovir Intravenous Infusion is a sterile material consisting Test solution (b). Dilute 1 volume of test solution (a) to 10
of aciclovir sodium, prepared from Aciclovir with the aid of a volumes with 0.1 M sodium hydroxide.
68
IP 2007 ACICLOVIR TABLETS
Reference solution (a). A 0.05 per cent w/v solution of Aciclovir Tablets
aciclovir RS in 0.1 M sodium hydroxide.
Acyclovir Tablets
Reference solution (b). A 0.005 per cent w/v solution of
guanine in 0.1 M sodium hydroxide. Aciclovir Tablets contain not less than 95.0 per cent and not
more than 105.0 per cent of the stated amount of aciclovir,
Apply to the plate 10 µl of each solution. Allow the mobile
C8H11N5O3.
phase to rise 12 cm. Dry the pate in a current of warm air and
examine in ultraviolet light at 254 nm. Any secondary spot
Identification
corresponding to guanine in the chromatogram obtained with
test solution (a) is not more intense than the spot in the A. When examined in the range 230 nm to 360 nm (2.4.7), the
chromatogram obtained with reference solution (b) (1per cent). solution prepared in the Assay shows an absorption maximum
Related substances. Determine by thin-layer chromatography at about 255 nm and a broad shoulder at about 274 nm.
(2.4.17), coating the plate with silica gel GF254. B. In the test for Guanine, the principal spot in the
Mobile phase. A mixture of 80 volumes of dichloromethane, chromatogram obtained with test solution (b) corresponds to
20 volumes of methanol and 2 volumes of strong ammonia that in the chromatogram obtained with reference solution (a).
solution.
Tests
Prepare the following solutions immediately before use.
Guanine. Determine by thin-layer chromatography (2.4.17),
Test solution. Dissolve a suitable quantity of the substance coating the plate with cellulose F254. (such as Merck
under examination in dimethyl sulphoxide to produce a celllulose F plates).
solution containing 2.5 per cent of aciclovir.
Mobile phase. A mixture of 10 volumes of 1- propanol,
Reference solution. Dilute 1 volume of the test solution to 200 30 volumes of strong ammonia solution and 60 volumes of a
volumes with dimethyl sulphoxide. 5 per cent w/v solution of ammonium sulphate.
Apply to the plate 2 µl of each solution. Allow the mobile Test solution (a). Shake a quantity of the powdered tablets
phase to rise 10 cm. Dry the plate in a current of warm air and containing 0.25 g of Aciclovir with 25 ml of 0.1 M sodium
examine in ultraviolet light 254 nm. Any secondary spot with hydroxide for 10 minutes. Add a sufficient quantity of 0.1 M
an Rf value greater than that of the principal spot in the sodium hydroxide to produce 50 ml. Allow to stand and allow
chromatogram obtained with the test solution is not more any undissolved material to settle before application to the
intense than the spot in the chromatogram obtained with plate.
reference solution (0.5 per cent).
Test solution (b). Dilute 1 volume of test solution (a) to
Bacterial endotoxins (2.2.3). Not more than 0.174 Endotoxin 10 volumes with 0.1 M sodium hydroxide.
Units per mg of acyclovir.
Assay. Dissolve a quantity of the mixed contents of 10 aciclovir RS in 0.1 M sodium hydroxide.
containers containing 0.10 g of Aciclovir in sufficient 0.1 M
hydrochloric acid to produce 500.0 ml. Dilute 5.0 ml of the Reference solution (b). A 0.005 per cent w/v solution of
resulting solution to 100.0 ml with 0.1 M hydrochloric acid. guanine in 0.1 M sodium hydroxide.
Measure the absorbance of the resulting solution at the Apply to the plate 10 µl of each solution. Allow the mobile
maximum at about 255 nm (2.4.7). Calculate the content of phase to rise 12 cm. Dry the plate in a current of warm air and
C8H11N5O3 taking 560 as the specific absorbance at 255 nm. examine in ultraviolet light at 254 nm. Any secondary spot
Storage. Store protected from moisture, in a sterile, tamper- corresponding to guanine in the chromatogram obtained with
evident container sealed so as to exclude micro-organisms, at test solution (a) is not more intense than the spot in the
a temperature not exceeding 30°. chromatogram obtained with reference solution (b) (1 per
cent).
Labelling. The label states (1) the quantity of aciclovir sodium
in the sealed container in terms of the equivalent amount of Related substances. Determine by thin-layer chromatography
Aciclovir; (2) the strength of the constituted solution in terms (2.4.17), coating the plate with silica gel GF254.
of the equivalent amount of Aciclovir in a suitable dose-volume. Mobile phase. A mixture of 80 volumes of dichloromethane,
20 volumes of methanol and 2 volumes of strong ammonia
solution.
69
ADENINE IP 2007
Test solution. Shake a quantity of the powdered tablets B. In the test for Related substances, the principal spot in the
containing 0.25 g of Aciclovir with 10 ml of dimethyl sulphoxide chromatogram obtained with test solution (b) corresponds to
for 15 minutes and filter. that in the chromatogram obtained with reference solution (a).
Reference solution. Dilute 0.7 volume of the test solution to C. To 1 g add 3.5 ml of propionic anhydride, boil for 15 minutes
100 volumes with dimethyl sulphoxide. with stirring and cool. To the resulting crystalline mass add 15
ml of light petroleum (50° to 70°) and heat to boiling with
vigorous stirring. Cool and filter. Wash the precipitate with
phase to rise 10 cm. Dry the plate in a current of warm air and
two quantities, each of 5 ml, of light petroleum (50° to 70°).
examine in ultraviolet light at 254 nm. Any secondary spot
Dissolve the precipitate in 10 ml of water and boil for 1 minute.
with an Rf value greater than that of the principal spot in the
Allow the mixture to cool to 30° to 40° and filter. Dry the
precipitate at 105° for 1 hour. The precipitate melts at 237° to
intense than the spot in the chromatogram obtained with the
241° (2.4.21).
reference solution (0.7 per cent).
Other tests. Comply with the tests stated under Tablets. Tests
Assay. Weigh and powder 20 tablets. Weigh accurately a Appearance of solution. A 1.0 per cent w/v solution in dilute
quantity of the powder containing about 0.1g of Aciclovir, hydrochloric acid is clear (2.4.1) and colourless (2.4.1).
add 60 ml of 0.1 M sodium hydroxide and disperse with the
aid of ultrasound for 15 minutes. Add sufficient quantity of Related substances. Determine by thin-layer chromatography
0.1 M sodium hydroxide to produce 100.0 ml, mix well and (2.4.17), coating the plate with silica gel GF254.
filter. To 10.0 of the filtrate add 50 ml of water, 5.8 ml of 2 M Mobile phase. A mixture of 40 volumes of 1-propanol, 40
hydrochloric acid and sufficient water to produce 100.0 ml. volumes of ethyl acetate and 20 volumes of strong ammonia
To 5.0 ml of the resulting solution add sufficient 0.1 M solution.
hydrochloric acid to produce 50.0 ml and mix well. Measure
Test solution (a). Dissolve 0.1 g of the substance under
the absorbance of the solution at the maximum at about
examination in dilute acetic acid, with heating if necessary,
255 nm (2.4.7), using 0.1 M hydrochloric acid as the blank.
and dilute to 10 ml with the same solvent.
Calculate the content of C8H11N5O3 taking 560 as the specific
absorbance at 255 nm. Test solution (b). Dilute 1 ml of test solution (a) to 10.0 ml with
dilute acetic acid.
Reference solution (a). Dissolve 10 mg of adenine RS in dilute
acetic acid, with heating if necessary, and dilute to 10 ml with
the same solvent.
Adenine
Reference solution (b). Dilute 1 ml of test solution (b) to 20 ml
NH2 with dilute acetic acid.
N Reference solution (c). Dissolve 10 mg of adenine RS and 10

HN mg of adenosine RS in dilute acetic acid with heating if
N necessary and dilute to 10 ml with the same solvent.
N
C5H5N5 Mol. Wt. 135.1 phase to rise 12 cm. Dry in a current of warm air and examine in
Adenine is 1H-purin-6-amine. ultraviolet light at 254 nm. Any secondary spot in the
chromatogram obtained with test solution (a) is not more
Adenine contains not less than 98.5 per cent and not more
intense than the spot in the chromatogram obtained with
than 101.0 per cent of C5H5N5, calculated on the dried basis.
reference solution (b) (0.5 per cent). The test is not valid unless
Description. A white, crystalline powder; odourless. the chromatogram obtained with reference solution (c) shows
two clearly separated spots.
Identification
Test A may be omitted if tests B and C are carried out. Test B heavy metals, Method B (10 ppm).
may be omitted if tests A and C are carried out.
Chlorides (2.3.12). Suspend 2.5 g in 50 ml of water and boil for
A. Determine by infrared absorption spectrophotometry (2.4.6). 3 minutes. Cool and dilute to 50 ml with water and filter. To 10
Compare the spectrum with that obtained with adenine RS or ml of the filtrate (solution A) add 1 ml of strong ammonia
with the reference spectrum of adenine. solution and 3 ml of dilute silver nitrate solution. Filter, wash
70
IP 2007 ADRENALINE
the precipitate with a little quantity of water and dilute the an absorption maximum at about 280 nm; absorbance at about
filtrate to 15 ml with water. The resulting solution complies 280 nm, about 0.45.
with the limit test for chlorides (100 ppm).
C. To 1 ml of a neutral or faintly acid solution add dropwise a
Sulphates (2.3.17). Dilute 10 ml of solution A to 15 ml with 0.25 per cent w/v solution of ferric chloride until an emerald-
water. The solution complies with the limit test for sulphates green colour is produced. Add sodium bicarbonate solution
(300 ppm). gradually; the solution changes first to blue and then to red.
Sulphated ash (2.3.18). Not more than 0.1 per cent. D. To 1 ml of a 0.1 per cent w/v solution add 1 ml of a 1.0 per
Loss on drying (2.4.19). Not more than 0.5 per cent, determined cent v/v solution of 2,5-diethoxytetrahydrofuran in glacial
on 1.0 g by drying in an oven at 105°. acetic acid. Heat at 80° for 2 minutes, cool in ice and add 3 ml
of a 2.0 per cent w/v solution of 4-dimethylaminobenzaldehyde
Assay. Weigh accurately about 0.1 g, dissolve in a mixture of in a mixture of 19 volumes of glacial acetic acid and 1 volume
30 ml of anhydrous glacial acetic acid and 20 ml of acetic of hydrochloric acid. Mix and allow to stand for 2 minutes.
anhydride. Titrate with 0.1 M perchloric acid, determining The solution becomes yellow and is similar to the one obtained
the end-point potentiometrically (2.4.25). Carry out a blank by performing the test in the same manner but omitting the
titration. substance under examination (distinction from noradrenaline).
1 ml of 0.1 M perchloric acid is equivalent to 0.01351 g of
C5H5N5. Tests
Storage. Store protected from light and moisture. Specific optical rotation (2.4.22). –50.0° to –53.5°, determined
in a freshly prepared 4.0 per cent w/v solution in 1 M
hydrochloric acid.
Phenones. Absorbance of a 0.2 per cent w/v solution in 0.1 M
Adrenaline hydrochloric acid at the maximum at about 310 nm, not greater
Epinephrine than 0.20, calculated on the dried basis (2.4.7).
Noradrenaline. Dissolve 5 mg in 1 ml of a 0.5 per cent w/v
OH H solution of tartaric acid, add 4 ml of buffer pH 9.6, mix, add 1
HO N ml of a freshly prepared 0.5 per cent w/v solution of sodium
CH3 1,2-naphthaquinone-4-sulphonate, mix and allow to stand
for 30 minutes. Add 0.2 ml of a 1 per cent v/v solution of
HO
benzalkonium chloride solution, mix, add 15 ml of toluene
previously washed with buffer pH 9.6 and filtered through a
C9H13NO3 Mol. Wt. 183.3 dry filter paper, shake for 30 minutes and allow to separate,
Adrenaline is (R)-1-(3,4-dihydroxyphenyl)-2- centrifuging if necessary. Any red or purple colour in the
methylaminoethanol toluene layer is not more intense than that produced by
treating a solution of 0.40 mg of noradrenaline acid tartrate
Adrenaline contains not less than 98.5 per cent and not more
and 9 mg of noradrenaline-free adrenaline acid tartrate in 1
than 101.0 per cent of C9H13NO3, calculated on the dried basis.
ml of water in a similar manner.
Description. A white or creamy-white, microcrystalline powder
or granules. It gradually darkens on exposure to light and air, Sulphated ash (2.3.18). Not more than 0.1 per cent.
decomposition being faster in the presence of moisture and at Loss on drying (2.4.19). Not more than 1.0 per cent, determined
higher temperatures. on 1.0 g by drying over phosphorus pentoxide at a pressure
not exceeding 0.7 kPa for 18 hours.
Identification
Test A may be omitted if tests B, C and D are carried out. Test anhydrous glacial acetic acid, warming slightly, if necessary,
C may be omitted if tests A, B and D are carried out. to effect solution. Titrate with 0.1 M perchloric acid, using
crystal violet solution as indicator. Carry out a blank titration.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with adrenaline RS 1 ml of 0.1 M perchloric acid is equivalent to 0.01832 g of
or with the reference spectrum of adrenaline. C9H13NO3.
B. When examined in the range 230 nm to 360 nm (2.4.7), a Storage. Store protected from light in containers preferably
0.003 per cent w/v solution in 0.01 M hydrochloric acid shows filled with nitrogen.
71
ADRENALINE TARTRATE IP 2007
Adrenaline Tartrate pH (2.4.24). 2.8 to 4.0, determined in a 1.0 per cent w/v solution.
Phenones. Absorbance of a 0.2 per cent w/v solution in 0.1 M
Adrenaline Acid Tartrate; Adrenaline Bitartrate;
hydrochloric acid at the maximum at about 310 nm, not more
Epinephrine Bitartrate. than 0.10, calculated on the dried basis (2.4.7).
Noradrenaline. Determine by thin-layer chromatography
OH H OH (2.4.17), coating the plate with silica gel G.
H
HO N COOH
CH3 , HOOC Mobile phase. A mixture of 100 volumes of acetone, 100
OH volumes of dichloromethane and 1 volume of anhydrous
HO H formic acid.
Test solution. Dissolve 2.5 g of the substance under
C9H13NO3,C4H6O6 Mol. Wt. 333.3
examination in 100 ml of water.
Adrenaline tartrate is (R)-1-(3,4-dihydroxyphenyl)-2-
Reference solution (a). A freshly prepared 0.125 per cent w/v
methylaminoethanol hydrogen tartrate.
solution of noradrenaline bitartrate RS in water.
Adrenaline Tartrate contains not less than 98.0 per cent and Reference solution (b). A freshly prepared 0.025 per cent w/v
not more than 101.0 per cent of C9H13NO3,C4H6O6, calculated solution of noradrenaline bitartrate RS in water.
on the dried basis.
Reference solution (c). A mixture of equal volumes of the test
Description. A white or greyish-white, crystalline powder; solution and reference solution (b).
odourless. It darkens on exposure to air and light,
decomposition being faster in the presence of moisture and at Apply to the plate 6 µl of each of the test solution and reference
higher temperatures. solutions (a) and (b) and 12 µl of reference solution (c) as
bands 20 mm by 2 mm.
Identification Allow the applied bands to dry and spray them with a saturated
Dissolve about 1 g in 10 ml of water containing 0.1 g of sodium solution of sodium bicarbonate. Allow the plate to dry in air,
metabisulphite, add a slight excess of dilute ammonia solution spray the applied bands twice with acetic anhydride, drying
and allow to stand at about 4° for 1 hour, filter and reserve the the plate between the two sprayings and heat the plate at 50°
filtrate for test D. Wash the precipitate with three successive for 90 minutes and develop the chromatograms. After removal
quantities, each of 2 ml, of cold water, then with 5 ml of cold of the plate, allow it to dry in air and spray with a freshly
ethanol (95 per cent) and finally with 5 ml of cold ether and prepared mixture of 8 volumes of methanol, 2 volumes of
dry over silica gel at a pressure of 1.5 to 2.5 kPa for 3 hours. ethylenediamine and 2 volumes of a 0.5 per cent w/v solution
The residue comply with the following tests. of potassium ferricyanide. Dry the plate at 60° for 10 minutes
and examine in ultraviolet light at 254 nm and 365 nm. Any
A. Determine by infrared absorption spectrophotometry (2.4.6). band situated between the two most intense bands in the
Compare the spectrum with that obtained with adrenaline RS chromatogram obtained with the test solution is not more
or with the reference spectrum of adrenaline. intense than the corresponding band in the chromatogram
B. When examined in the range 230 nm to 360 nm (2.4.7), a obtained with reference solution (b). The test is not valid
0.005 per cent w/v solution in 0.01 M hydrochloric acid shows unless the chromatogram obtained with reference solution (c)
an absorption maximum only at about 279 nm; absorbance at exhibits between the two most intense bands a clearly
about 279 nm, about 0.4. separated band corresponding to the most intense band in
the chromatogram obtained with reference solution (a).
C. The filtrate reserved above gives the reactions of tartrates
(2.3.1). Sulphated ash (2.3.18). Not more than 0.1 per cent.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Tests on 1.0 g by drying over phosphorus pentoxide at a pressure
Appearance of solution. A 5.0 per cent w/v solution examined of 1.5 to 2.5 kPa for 18 hours.
immediately after preparation is not more opalescent than Assay. Weigh accurately about 0.3 g and dissolve in 50 ml of
opalescence standard OS2 (2.4.1) and not more intensely anhydrous glacial acetic acid, warming slightly, if necessary,
coloured than reference solution BYS4 (2.4.1). to effect solution. Titrate with 0.1 M perchloric acid, using
Specific optical rotation (2.4.22). –50.0° to –54.0 °, determined crystal violet solution as indicator. Carry out a blank titration.
in a freshly prepared 4.0 per cent w/v solution in 1 M 1 ml of 0.1 M perchloric acid is equivalent to 0.03333 g of
hydrochloric acid. C9H13NO3,C4H6O6.
72
IP 2007 ADRENALINE INJECTION
Storage. Store protected from light in containers preferably Chromatographic system

filled with nitrogen. – a stainless steel column 10 cm x 4.6 mm, packed with
octadecylsilane chemically bonded to porous silica or
ceramic microparticles 5 to 10 µm (such as Nucleosil
ODS),
Adrenaline Injection – mobile phase: Dissolve 4.0 g of tetramethylammonium
Adrenaline Bitartrate Injection; Adrenaline Acid Tartrate hydrogen sulphate, 1.1 g of sodium heptanesulphonate
Injection; Adrenaline Tartrate Injection; Epinephrine Tartrate and 2 ml of 0.1 M disodium edetate in 1000 ml of 5 per
Injection cent v/v solution of methanol, with pH adjusted to 3.5
to 3.6 with 1 M sodium hydroxide,
Adrenaline Injection is a sterile, isotonic solution containing – flow rate. 2 ml per minute,
0.18 per cent w/v of Adrenaline Tartrate in Water for Injection. – spectrophotometer set at 205 nm,
Adrenaline Injection contains the equivalent of not less than – a 20 µl loop injector.
0.09 per cent and not more than 0.115 per cent w/v of adrenaline,
Inject reference solution (b). The test is not valid unless the
C9H13NO3.
resolution factor between the two principal peaks is not less
Description. A clear, colourless or almost colourless solution. than 2.0.
Identification Inject the test solution and reference solution (a). The area of
any peak corresponding to noradrenaline is not greater than
A. To an appropriate quantity add sufficient 0.01M the area of the principal peak in the chromatogram obtained
hydrochloric acid to produce a solution containing 0.005 per with reference solution (a).
cent w/v of adrenaline. When examined in the range 230 nm to
Other tests. Complies with the tests stated under Parenteral
360 nm (2.4.7), the solution shows an absorption maximum at
Preparations (Injections).
about 279 nm; absorbance at about 279 nm, about 0.4.
Assay. Extract 30.0 ml in a separator with three quantities,
B. To 1 ml add dropwise a 0.25 per cent w/v solution of ferric
each of 25 ml, of carbon tetrachloride, shaking vigorously
chloride until an emerald-green colour is produced. Add
for 1 minute each time; reject the carbon tetrachloride extracts.
sodium bicarbonate solution gradually; the solution changes
Add 0.2 ml of starch solution and, dropwise with swirling, a
first to blue and then to red.
solution prepared by dissolving 0.5 g of iodine and 1.5 g of
C. To 10 ml add 2 ml of disodium hydrogen phosphate solution potassium iodide in 25 ml of water, until the blue colour
and sufficient iodine solution to produce a brown colour. persists. Immediately add just sufficient 0.1 M sodium
Add 0.1 M sodium thiosulphate dropwise until excess iodine thiosulphate to discharge the blue colour and proceed further
is removed; a red colour is produced. without delay. Add 2.1 g of sodium bicarbonate and swirl
until most of the sodium bicarbonate has dissolved. Using a
Tests syringe, rapidly inject 1.0 ml of acetic anhydride directly into
the contents of the separator, insert the stopper, and shake
Appearance of solution. Examine the injection in a clear glass
test-tube against a white background; it is not pinkish and vigorously until the evolution of carbon dioxide ceases (7 to
does not contain a precipitate. If any yellow colour is observed, 10 minutes), releasing the pressure when necessary through
it is not more intense than a reference solution prepared by the stop-cock. Allow to stand for 5 minutes and extract with
diluting 0.4 ml of 0.1 M iodine to 100 ml with water, when six successive quantities, each of 25 ml, of chloroform, filtering
viewed similarly. each extract into a beaker through a small plug of cotton wool
moistened with chloroform. Remove the chloroform, heat the
pH (2.4.24). 2.8 to 3.6. residue at 105° for 30 minutes, allow to cool and weigh. Dissolve
Noradrenaline. Determine by liquid chromatography (2.4.14). the residue in 5.0 ml of chloroform, swirling to assist solution,
and determine the specific optical rotation of the resulting
Test solution. Substance under examination.
solution (2.4.22), using a 2-dm tube. Calculate the percentage
Reference solution (a). A 0.0018 per cent w/v solution of content of C9H13NO3, in the injection from the expression 1.974
noradrenaline acid tartrate in the mobile phase. W (0.5 + 0.5 R/93), where W is the weight of the residue in g
Reference solution (b). A solution containing 0.0018 per cent and R is its specific optical rotation (in degrees without regard
w/v of noradrenaline-free adrenaline acid tartrate and to the sign).
0.0018 per cent w/v of noradrenaline acid tartrate in the Storage. Store protected form light, in a single dose or multiple
mobile phase. dose container.
73
ALBENDAZOLE IP 2007
Labelling. The label states (1) the quantity of active ingredient Apply to the plate 10 µl of each solution. After development,
in parts per 1000 or mg per ml in terms of equivalent amount of dry in a current of warm air and examine in ultraviolet light at
adrenaline; (2) that the injection should not be used if it is 254 nm. Any secondary spot in the chromatogram obtained
pinkish or darker than slightly yellow. with test solution (a) is not more intense than the principal
spot in the chromatogram obtained with reference solution
(a).
Albendazole heavy metals, Method B (10 ppm)
H O CH3 Sulphated ash (2.3.18). Not more than 0.2 per cent.
N O Loss on drying (2.4.19). Not more than 0.5 per cent, determined
NH on 1.0 g by drying in an oven at 105° for 4 hours.
H3C N
S
C12H15N3O2S Mol. Wt. 265.3
acid, using crystal violet solution as indicator. Carry out a
Albendazole is methyl 5-propylthio-1H-benzimidazol-2-yl- blank titration.
carbamate.
Albendazole contains not less than 98.0 per cent and not C12H15N3O2S
more than 102.0 per cent of C12H15N3O2S, calculated on the
dried basis.
Description. A white to pale buff-coloured powder.
Identification
Albendazole Tablets
Test A may be omitted if tests B and C are carried out. Test B
may be omitted if tests A and C are carried out. Albendazole Tablets contain Albendazole. The tablets may
contain permitted flavouring agents.
Compare the spectrum with that obtained with albendazole Albendazole Tablets contain not less than 92.5 per cent and
RS or with the reference spectrum of albendazole. not more than 107.5 per cent of the stated amount of
albendazole, C12H15N3O2S.
B. In the test for Related substances, the principal spot in the
chromatogram obtained with test solution (b) corresponds to Identification
that in the chromatogram obtained with reference solution
(b). A. Determine by thin-layer chromatography (2.4.17), coating
C. Melting point (2.4.21). 208° to 210°. the plate with silica gel GF254.
Mobile phase. A mixture of 60 volumes of chloroform, 10
Tests volumes of ether and 10 volumes of glacial acetic acid.
Related substances. Determine by thin-layer chromatography Test solution. Add a quantity of the powdered tablets
(2.4.17), coating the plate with silica gel GF254. containing 200 mg of Albendazole to 20 ml of a mixture of 18
Mobile phase. A mixture of 60 volumes of chloroform, 10 volumes of chloroform and 1 volume of formic acid, warm the
volumes of ether and 10 volumes of glacial acetic acid. suspension on a water-bath for 15 minutes, cool and filter.
Dilute 10 ml of the filtrate with an equal volume of glacial
Test solution (a). Dissolve 0.2 g of the substance under acetic acid.
examination in sufficient glacial acetic acid to produce 10 ml.
Reference solution. Dissolve 25 mg of albendazole RS in
Test solution (b). Dilute 1 ml of test solution (a) to 4 ml with sufficient glacial acetic acid to produce 5 ml.
glacial acetic acid.
Apply to the plate 10 µl of each solution. After development,
Reference solution (a). Dilute 1 ml of test solution (a) to
dry the plate in a current of warm air and examine in ultraviolet
200 ml with glacial acetic acid.
Reference solution (b). Dissolve 25 mg of albendazole RS in obtained with the test solution corresponds to that in the
sufficient glacial acetic acid to produce 5 ml. chromatogram obtained with the reference solution.
74
IP 2007 ALGINIC ACID
B. Extract a quantity of the powdered tablets containing 100 Tests

mg of Albendazole with 100 ml of 0.1 M methanolic
hydrochloric acid, filter and dilute 1 ml of the filtrate to 100 ml pH (2.4.24). 1.5 to 3.5, determined in a 3.0 per cent w/v
with 0.1 M sodium hydroxide. The absorbance of the resulting dispersion in water.
solution at the maximum at about 309 nm, about 0.74 (2.4.7). Arsenic (2.3.10). Mix 2.0 g with 5 ml of sulphuric acid, add a
few glass beads and digest at a temperature not exceeding
Tests 120° until charring begins. Additional sulphuric acid may be
Other tests. Comply with the tests stated under Tablets. added if necessary but the total volume of acid added should
not exceed 10 ml. Add cautiously, dropwise, hydrogen peroxide
Assay. Weigh and powder 20 tablets. Weigh accurately a solution (100 vol) allowing the reaction to subside and again
quantity of the powder containing about 0.1 g of Albendazole, heating between addition of drops. Discontinue heating if
add about 150 ml 0.1 M methanolic hydrochloric acid, shake foaming becomes excessive. When the reaction has abated,
for 15 minutes and dilute to 250.0 ml with 0.1 M methanolic heat cautiously rotating the flask occasionally. Maintain
hydrochloric acid. Mix, filter and dilute 5.0 ml of the filtrate to oxidising conditions at all times during the digestion by adding
250.0 ml with 0.1 M sodium hydroxide. Measure the small quantities of the hydrogen peroxide solution whenever
absorbance of the resulting solution at the maximum at about the mixture turns brown or darkens. Continue the digestion
309 nm (2.4.7). Calculate the content of C12H15N3O2S taking until the organic matter has been destroyed, gradually raising
742 as the specific absorbance at 309 nm. the temperature until fumes of sulphur trioxide are copiously
Storage. Store protected from light. evolved and the solution becomes colourless or has only a
light straw colour. Cool, add cautiously 10 ml of water, mix,
and again evaporate till there is strong fuming, repeating this
procedure to remove any trace of hydrogen peroxide. Cool,
Alginic Acid add cautiously 10 ml of water, wash the sides of the flask with
a few ml of water and dilute with water to 35 ml. The resulting
Polymannuronic Acid
solution complies with the limit test for arsenic (5 ppm).
Alginic acid is a hydrophilic colloidal mixture of polyuronic
acids, [(C6H8O6) n], composed of residues of D-mannuronic
heavy metals, Method B (40 ppm). Use nitric acid Sp. in
acid and L-guluronic acid extracted with dilute alkali from
place of sulphuric acid Sp. to wet the sample.
various species of brown seaweeds (Fam. Phaeophyceae).
Acid value. Not less than 230, calculated on the dried basis
Alginic Acid contains not less than 19.0 per cent and not more
and determined in the following manner. Weigh accurately
than 25.0 per cent of carboxylic acid groups (COOH), calculated
about 1.0 g and suspend in a mixture of 50 ml of water and 30
on the dried basis.
ml of a 4.4 per cent w/v solution of calcium acetate. Shake
Description. A white to yellowish-white, fibrous powder; vigorously, allow the mixture to stand for 1 hour, add
odourless. phenolphthalein solution and titrate the liberated acetic acid
with 0.1 M sodium hydroxide. Carry out a blank titration.
Identification
Calculate the acid value from the expression 5.611 A/W, where
A. To 5 ml of a 0.75 per cent w/v solution in 0.1 M sodium A is the volume, in ml, of 0.1 M sodium hydroxide consumed
hydroxide add 1 ml of calcium chloride solution; a gelatinous and W is the weight, in g, of the sample.
precipitate is formed. Microbial contamination (2.2.9). 1 g is free from Escherichia
B. To 5 ml of the solution obtained in test A add 1 ml of 2 M coli and 10 g is free from salmonellae.
sulphuric acid; a gelatinous precipitate is formed. Total ash (2.3.19). Not more than 4.0 per cent, determined on
C. To about 5 mg in a test-tube add 5 ml of water, 1 ml of a 0.5 g by Method B.
freshly-prepared 1 per cent w/v solution of 1,3- Loss on drying (2.4.19). Not more than 15.0 per cent, determined
naphthalenediol in ethanol (95 per cent) and 5 ml of on 0.1 g by drying in an oven at 105° for 4 hours.
hydrochloric acid. Heat the mixture to boiling, boil gently for
Assay. Weigh accurately about 0.25 g, add 25 ml of water and
3 minutes and cool to about 15°. Transfer the contents of the
25.0 ml of 0.1 M sodium hydroxide and titrate with 0.1 M
test-tube to a small separator with the aid of 5 ml of water and
hydrochloric acid using 0.2 ml of dilute phenolphthalein
extract with 15 ml of di-isopropyl ether; the di-isopropyl ether
solution as indicator.
extract exhibits a deep purple colour which is more intense
than that exhibited by a blank prepared in the same manner 1 ml of 0.1 M sodium hydroxide is equivalent to 0.004502 g of
without the substance under examination. carboxylic acid groups (COOH).
75
ALLOPURINOL IP 2007
Allopurinol Test solution. A 2.5 per cent w/v solution of the substance
under examination in strong ammonia solution.
OH Reference solution. A 0.005 per cent w/v solution of 5-
aminopyrazole-4-carboxamide hemisulphate RS in strong
N ammonia solution.
N
N N Apply to the plate 10 µl of each solution. After development,
H dry the plate in a current of warm air and examine in ultraviolet
C5H4N4O Mol. Wt. 136.1 light at 254 nm. Any secondary spot in the chromatogram
obtained with the test solution is not more intense than the
Allopurinol is a tautomeric mixture of 1H-pyrazolo[3,4-d] spot in the chromatogram obtained with the reference solution.
pyrimidin-4-ol and 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-
4-one. Heavy metals (2.3.13). Mix carefully 1.0 g in a silica crucible
with 4 ml of a 25 per cent w/v solution of magnesium sulphate
Allopurinol contains not less than 98.0 per cent and not more in 1 M sulphuric acid and heat cautiously to dryness. Ignite
than 101.0 per cent of C5H4N4O, calculated on the dried basis. the residue at a temperature not exceeding 800° and continue
Description. A white or almost white, crystalline powder. heating until a white or greyish residue is obtained. Allow to
cool, moisten with 0.2 ml of 1 M sulphuric acid, evaporate,
Identification ignite again and allow to cool. The total ignition period should
Test A may be omitted if tests B, C and D are carried out. Tests be less than 2 hours. Dissolve the residue with two quantities,
B, C and D may be omitted if test A is carried out. each of 5 ml, of 2 M hydrochloric acid. Add 2 drops of dilute
phenolphthalein solution and strong ammonia solution
A. Determine by infrared absorption spectrophotometry (2.4.6). dropwise until a pink colour is produced. Cool, add glacial
Compare the spectrum with that obtained with allopurinol acetic acid until the solution gets decolorised and add a
RS. further 0.5 ml. Filter, if necessary, and dilute the solution to 20
B. Dissolve 0.1 g in 10 ml of 0.1 M sodium hydroxide and add ml with water. The resulting solution complies with the limit
sufficient 0.1 M hydrochloric acid to produce 100.0 ml; dilute test for heavy metals, Method D (20 ppm).
10.0 ml to 100.0 ml with 0.1 M hydrochloric acid and dilute Sulphated ash (2.3.18). Not more than 0.1 per cent.
10.0 ml of this solution to 100.0 ml with 0.1 M hydrochloric
acid. When examined in the range 230 nm to 360 nm (2.4.7), Loss on drying (2.4.19). Not more than 0.5 per cent, determined
the resulting solution shows an absorption maximum at about on 1.0 g by drying in an oven at 105°.
250 nm and a minimum at about 231 nm; ratio of the absorbance Assay. Weigh accurately about 0.2 g and dissolve with gentle
at the minimum at about 231 nm to that at the maximum at heating, if necessary, in 50 ml of dimethylformamide. Titrate
about 250 nm, 0.52 to 0.62. with 0.1 M tetrabutylammonium hydroxide, determining the
C. Dissolve 50 mg in 5 ml of dilute sodium hydroxide solution, end-point potentiometrically (2.4.25). Carry out a blank titration.
add 1 ml of alkaline potassium mercuri-iodide solution, heat 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
to boiling and allow to stand; a flocculent yellow precipitate is 0.01361 g of C5H4N4O.
produced.
D. Shake about 0.1 g with 5 ml of dilute sodium hydroxide
solution, add 3 ml of lithium and sodium molybdo- Allopurinol Tablets
phosphotungstate solution and 5 ml of a 20 per cent w/v
solution of sodium carbonate; a grey-blue colour is produced. Allopurinol Tablets contain not less than 92.5 per cent and
Tests allopurinol, C5H4N4O.
Appearance of solution. A 5.0 per cent w/v solution in 2 M
Identification
sodium hydroxide is clear, (2.4.1), and not more intensely
coloured than reference solution YS6 or GYS4 (2.4.1). A. When examined in the range 230 nm to 360 nm (2.4.7), the
Related substances. Determine by thin-layer chromatography solution obtained in the Assay shows an absorption maximum
(2.4.17), coating the plate with silica gel GF254. only at about 250 nm.
Mobile phase. A mixture of 60 volumes of 2-butanone, 20 B. Shake a quantity of the powdered tablets containing about
volumes of 2-methoxyethanol and 20 volumes of strong 0.1 g of Allopurinol with 5 ml of dilute sodium hydroxide
ammonia solution. solution, add 3 ml of lithium and sodium molybdo-
76
IP 2007 ALOES
phosphotungstate solution and 5 ml of a 20 per cent w/v Description. Unground Curacao Aloes — Brownish-black,
solution of sodium carbonate; a grey-blue colour is produced. opaque masses; fractured surface uneven, waxy and somewhat
resinous; odour, strong and characteristic.
Tests
Unground Cape Aloes — Dark-brown or greenish-brown to
Related substances. Determine by thin-layer chromatography olive-brown masses; fractured surface shiny and conchoidal;
(2.4.17), coating the plate with silica gel GF254. odour, strong and characteristic.
Mobile phase. A mixture of 60 volumes of 2-butanone, 20 Identification
volumes of 2-methoxyethanol and 20 volumes of strong
Mix 0.5 g with 50 ml of water, boil until nearly dissolved, cool,
ammonia solution.
add 0.5 g of silica gel and filter. On the filtrate carry out the
Test solution. Shake a quantity of the powdered tablets following tests.
containing about 0.25 g of Allopurinol with 10 ml of strong
A. Heat 5 ml with 0.2 g of borax until dissolved, add a few
ammonia solution and filter.
drops of this solution to a test-tube nearly filled with water; a
Reference solution. A 0.005 per cent w/v solution of 5- green fluorescence is produced.
aminopyrazole-4-carboxamide hemisulphate RS in strong
B. Mix 2 ml with 2 ml of bromine water; a pale yellow precipitate
ammonia solution
is produced. The supernatant liquid is violet with Curacao
Apply to the plate 10 µl of each solution. After development, Aloes; no such violet colour appears with Cape Aloes.
dry the plate in a current of warm air and examine in ultraviolet C. Mix 5 ml with 2 ml of nitric acid; with Cape Aloes a reddish-
light at 254 nm. Any secondary spot in the chromatogram yellow colour is produced; with Socotrine Aloes a pale
obtained with the test solution is not more intense than the brownish-yellow colour is produced; with Cape Aloes a
spot in the chromatogram obtained with the reference solution. yellowish-brown colour passing rapidly to green is produced.
Disintegration (2.5.1). 30 minutes. D. Determine by thin-layer chromatography (2.4.17), coating
Other tests. Comply with the tests stated under Tablets. the plate with silica gel G.
Assay. Weigh and powder 20 tablets. Weigh accurately a Mobile phase. A mixture of 100 volumes of ethyl acetate,
quantity of the powder containing about 0.1 g of Allopurinol 17 volumes of methanol and 13 volumes of water.
and shake with 20 ml of 0.05 M sodium hydroxide for 15 to 20 Test solution. Heat 0.5 g, in powder, with 20 ml methanol to
minutes, add 75 ml of 0.1 M hydrochloric acid shake for 10 boiling on a water-bath, shake well, decant the supernatant
minutes, add sufficient 0.1 M hydrochloric acid to produce liquid, keep at 4° and use within 24 hours.
250.0 ml, filter and dilute 5.0 ml of the filtrate to 250.0 ml with Reference solution. Dissolve 50 mg of barbaloin in 10 ml
0.1 M hydrochloric acid. Measure the absorbance of the methanol.
resulting solution at the maximum at about 250 nm (2.4.7) using
0.1 M hydrochloric acid as the blank. Apply to the plate 5 µl of each solution as bands 20 mm x
3 mm. Allow the mobile phase to rise 15 cm. Dry the plate in a
Calculate the content of C5H4N4O, taking 563 as the specific current of air, spray with a 10 per cent w/v solution of potassium
absorbance at 250 nm. hydroxide in methanol and examine in ultraviolet light at 365
nm. The chromatogram obtained with the reference solution
shows a yellow band with an Rf value of 0.4 to 0.5. In the case
of Curacao Aloes, the chromatogram obtained with the test
Aloes solution shows a yellow fluorescent band corresponding to
that due to barbaloin in the chromatogram obtained with the
Aloes is the dried juice of the leaves of Aloe barbadensis
reference solution and in the lower part a light blue fluorescent
Miller (A. vera Linn), known in commerce as Curacao Aloes or
band (corresponding to aloesine). In the case of Cape Aloes,
Barbados Aloes, or of A. ferox Miller and hybrids of this species
the test solution shows a yellow fluorescent band
with A. africana Miller and A. spicata Baker, known in
corresponding to that due to barbaloin in the chromatogram
commerce as Cape Aloes (Fam. Liliaceae). Indian Aloes of
obtained with the reference solution and in the lower part two
commerce is obtained from A. barbadensis.
yellow fluorescent bands (due to aloinosides A and B) as well
Aloes contains not less than 50.0 per cent of water-soluble as a blue fluorescent band (due to aloesine). Heat the plate at
extractive. Curacao Aloes contains not less than 18.0 per cent 110° for 5 minutes. In the case of Curacao Aloes, with the test
and Cape Aloes not less than 28.0 per cent of solution a violet fluorescent band appears just below the yellow
hydroxyanthracene derivatives, calculated as anhydrous band corresponding to barbaloin while in the case of Cape
barbaloin. Aloes no such violet band appears.
77
ALPRAZOLAM IP 2007
Tests Alprazolam
Ethanol-insoluble substances. Weigh accurately about 1.0 g,
in fine powder, and add to 50 ml of ethanol (95 per cent) in a H3C N
flask. Reflux the mixture for 15 minutes. Remove the source of N
heat and set aside for 1 hour, shaking frequently, filter through N
a small dried and tared filter paper or suitable filtering crucible
and wash the residue on the filter with ethanol (95 per cent) N
Cl
till the washings are colourless. The residue after drying to
constant weight at 105° weighs not more than 0.1 g.
Water-soluble extractive. Weigh accurately about 2.0 g, in
fine powder, and macerate with about 60 to 70 ml of water in a
flask. Shake the mixture at 30-minute intervals for 8 hours and
C17H13ClN4 Mol. Wt. 308.8
allow to stand for a further 16 hours without shaking. Filter,
wash the flask and the residue with small portions of water, Alprazolam is 8-chloro-1-methyl-6-phenyl-4H-1,2,4-
passing the washings through the filter until the filtrate triazolo[4,3-a][1,4]benzodiazepine.
measures 100 ml. Evaporate 50 ml of this filtrate to dryness in Alprazolam contains not less than 98.0 per cent and not more
a tared dish on a water-bath and dry at 105° for 3 hours; the than 102.0 per cent of C17H13ClN4, calculated on the dried basis.
residue weighs not less than 0.5 g.
Description. A white to off-white, crystalline powder.
Total ash (2.3.19). Not more than 0.5 per cent, determined on
1.0 g by Method A. Identification
Loss on drying (2.4.19). Not more than 12 per cent, determined
on 1.0 g by drying in an oven at 105°.
Compare the spectrum with that obtained with alprazolam RS
Assay. Moisten 0.2 g, in fine powder, with 2 ml of methanol, or with the reference spectrum of alprazolam.
add 5 ml of water at about 60°, mix, add a further 75 ml of water
B. Dissolve 10.0 mg in water and dilute to 500.0 ml with the
at about 60°, shake for 30 minutes, cool, filter through a filter
same solvent. Dilute 20.0 ml of this solution to 100.0 ml with
paper, washing the flask with 20 ml of water and add sufficient
water. When examined in the range 210 nm to 360 nm (2.4.7),
water to the combined filtrate and washings to produce
the solution shows an absorption maximum at about 220 nm.
1000.0 ml. Transfer 10.0 ml of the solution to a flask containing
1 ml of a 60 per cent w/v solution of ferric chloride hexahydrate C. Melts at about 225° (2.4.21).
and 6 ml of hydrochloric acid, heat in a water-bath under a
reflux condenser for 4 hours so that the water level is always Tests
above that of the liquid in the flask, cool, transfer the solution
to a separating funnel, rinsing the flask successively with 4 ml Related substances. Determine by thin-layer chromatography
of 1 M sodium hydroxide and 4 ml of water and adding the (2.4.17), coating the plate with silica gel GF254.
rinsings to the contents of the separating funnel. Extract with Mobile phase. A mixture of 50 volumes of chloroform, 50
three quantities, each of 20 ml, of carbon tetrachloride and volumes of acetone, 50 volumes of ethyl acetate and 5 volumes
wash the combined carbon tetrachloride layers with two of methanol.
quantities, each of 100 ml, of water, discarding the washings.
Test solution. Dissolve 2 g in sufficient chloroform to produce
Dilute the organic phase to 100.0 ml with carbon tetrachloride,
50 ml.
evaporate 20.0 ml carefully to dryness on a water-bath and
dissolve the residue in 10.0 ml of 1 M sodium hydroxide. Reference solution. A 0.012 per cent w/v solution of
Immediately measure the absorbance of the resulting solution alprazolam RS in chloroform.
at the maximum at about 440 nm and at about 500 nm (2.4.7) Apply to the plate 10 µl of each solution. Allow the mobile
Calculate the content of anhydrous barbaloin, taking 200 as phase to rise 10 cm. Dry the plate in a current of air. Repeat the
the specific absorbance at 500 nm. The result of the Assay is development process a second time and examine in ultraviolet
not valid unless the ratio of the absorbance at about 500 nm to light at 254 nm. Any secondary spot in the chromatogram
that at about 440 nm is not less than 1.9. obtained with the test solution is not more intense than the
Storage. Store protected from light and moisture. spot in the chromatogram obtained with the reference solution.
Labelling. The label states whether the material is Curacao Heavy metals (2.3.13). 1.0 g complies with the limit test for
Aloes or Cape Aloes. heavy metals, Method B (20 ppm).
78
IP 2007 ALUMINIUM ACETATE EAR DROPS
Sulphated ash (2.3.18). Not more than 0.5 per cent. Reference solution. A solution containing 0.0025 per cent w/v
Loss on drying (2.4.19). Not more than 0.5 per cent, determined of alprazolam RS in acetonitrile.
on 1.0 g by drying in an oven at 60° at a pressure of 0.7 kPa for Chromatographic system
16 hours. – a stainless steel column 25 cm x 4.6 mm, packed with
porous silica particles, 5 to 10 µm in diameter,
– mobile phase: a mixture of 850 volumes of acetonitrile,
Test solution. Weigh accurately about 25 mg and dissolve in 80 volumes of chloroform, 50 volumes of 1-butanol, 20
sufficient acetonitrile to produce 100.0 ml. Dilute 10.0 ml of volumes of water and 0.5 volume of glacial acetic acid.
this solution to 100.0 ml with acetonitrile. – flow rate. 2 ml per minute,
Reference solution. A solution containing 0.0025 per cent w/v – spectrophotometer set at 254 nm,
of alprazolam RS in acetonitrile. – a 10 µl or 20 µl loop injector.
Chromatographic system Calculate the content of C17H13ClN4 in the tablet.

– a stainless steel column 25 cm x 4.6 mm, packed with Other tests. Comply with the tests stated under Tablets.
porous silica particles 3 to 10 µm,
80 volumes of chloroform, 50 volumes of 1-butanol, Test solution. Place 5 tablets in a flask, add 2 ml of water and
20 volumes of water and 0.5 volume of glacial acetic swirl to disperse the tablets. Add sufficient acetonitrile to
acid, produce 25.0 ml. Shake for 10 to 15 minutes and centrifuge if
– flow rate. 2 ml per minute, necessary. Dilute a portion of the clear solution with
– spectrophotometer set at 254 nm, acetonitrile to produce a solution containing 0.0025 per cent
– a 10 µl or 20 µl loop injector. w/v of alprazolam.
Inject the reference solution. The test is not valid unless the Reference solution. A 0.0025 per cent w/v solution of
relative standard deviation for replicate injections is not more alprazolam RS in acetonitrile.
than 2.0 per cent. Chromatographic system as described under Uniformity of
Inject alternately the test solution and the reference solution. content.
Calculate the percentage content of C17H13ClN4.
relative standard deviation for replicate injections is not
more than 2.0 per cent.
Inject alternately the test solution and the reference solution.
Alprazolam Tablets Calculate the content of C17H13ClN4 in the tablets.
Alprazolam Tablets contain not less than 90.0 per cent and Storage. Store protected from light.
alprazolam, C17H13ClN4
Identification Aluminium Acetate Ear Drops

In the Assay, the principal peak in the chromatogram obtained Aluminium Acetate Otic Drops; Aluminium Acetate
with the test solution corresponds to the peak in the Solution; Burow’s Solution.
Aluminium Sulphate 255 g
Tests Calcium Carbonate 100 g
Uniformity of content. Comply with the test stated under Tartaric Acid 45 g
Tablets. Glacial Acetic Acid 82.5 ml
Determine by liquid chromatography (2.4.14). Purified Water sufficient to produce 1000 ml
Test solution. Transfer one tablet to a container, add 0.4 ml of Dissolve the Aluminium Sulphate in 600 ml of Purified Water,
water on to the tablet, allow the tablet to stand for 2 minutes add Glacial Acetic Acid followed by Calcium Carbonate mixed
and swirl the container to disperse the tablet. Add sufficient with the remainder of the Purified Water and allow to stand for
acetonitrile to produce a solution containing 0.0025 per cent not less than 24 hours in a cool place, stirring occasionally.
w/v of alprazolam. Shake to mix and centrifuge, if necessary. Filter, add the Tartaric Acid to the filtrate and mix.
79
ALUMINIUM HYDROXIDE GEL IP 2007
Aluminium Acetate Ear Drops contain not less than 1.7 per water. The resulting solution complies with the limit test for
cent w/v and not more than 1.9 per cent w/v of aluminium, Al. heavy metals, Method A (10 ppm).
Description. A clear solution. Chlorides (2.3.12). Dissolve 0.5 g in 5 ml of dilute nitric acid,
boil, cool, dilute to 100 ml with water and filter. 20 ml of the
Tests filtrate complies with the limit test for chlorides (0.25 per cent).
Weight per ml (2.4.29). 1.06 g to 1.08 g. Sulphates (2.3.17). Dissolve 1.0 g in 5 ml of dilute hydrochloric
acid with the aid of heat. Cool and dilute to 100 ml with water.
Other tests. Comply with the tests stated under Ear Drops.
Mix well and filter, if necessary. To 5 ml of the filtrate add 2 ml
Assay. Dilute 10.0 ml to 100.0 ml with water. To 10.0 ml of the of dilute hydrochloric acid; the solution complies with the
resulting solution add 40.0 ml of 0.5 M disodium edetate, limit test for sulphates (0.3 per cent).
90 ml of water and 0.15 ml of methyl red solution. Neutralise
Neutralising capacity. Disperse 5.0 g in 100 ml of water, heat
by the addition of 1 M sodium hydroxide dropwise and warm
to 37°, add 100.0 ml of 0.1 M hydrochloric acid previously
on a water-bath for 30 minutes. Cool, add 1 ml of 2 M nitric
heated to 37°and stir continuously, maintaining the
acid and 5 g of hexamine and titrate with 0.05 M lead nitrate
temperature at 37°; the pH of the solution, at 37°, after 10, 15
using 0.5 ml of xylenol orange solution as indicator.
and 20 minutes, is not less than 1.8, 2.3 and 3.0 respectively
1 ml of 0.05 M disodium edetate is equivalent to 0.001349 g of and at no time is more than 4.5. Add 10.0 ml of 0.5 M
Al. hydrochloric acid previously heated to 37°, stir continuously
Storage. Store protected from light, in well-filled containers. for 1 hour maintaining the temperature at 37° and titrate with
0.1 M sodium hydroxide to pH 3.5.
Not more than 50.0 ml of 0.1 M sodium hydroxide is required.
Aluminium Hydroxide Gel Microbial contamination (2.2.9). Total viable aerobic count,
not more than 100 micro-organisms per ml, determined by plate
Aluminium Hydroxide Suspension; Aluminium Hydroxide count. 1 ml is free from Escherichia coli.
Mixture
Aluminium Hydroxide Gel is an aqueous suspension of hydrochloric acid by warming on a water-bath; cool to below
hydrated aluminium oxide together with varying quantities of 20° and dilute to 100.0 ml with water. To 20.0 ml of this solution,
basic aluminium carbonate and bicarbonate. It may contain add 40.0 ml of 0.05 M disodium edetate, 80 ml of water, and
Glycerin, Sorbitol, Sucrose or Saccharin as sweetening agents 0.15 ml of methyl red solution and neutralise by the dropwise
and Peppermint Oil or other suitable flavours. It may also addition of 1 M sodium hydroxide. Warm on a water-bath for
contain suitable antimicrobial agents. 30 minutes, add 3 g of hexamine and titrate with 0.05 M lead
Aluminium Hydroxide Gel contains not less than 3.5 per cent nitrate using 0.5 ml of xylenol orange solution as indicator.
and not more than 4.4 per cent w/w of Al2O3. 1 ml of 0.05 M disodium edetate is equivalent to 0.002549 g of
Description. A white, viscous suspension, translucent in thin Al2O3.
layers; small amounts of clear liquid may separate on standing. Storage. Store at a temperature not exceeding 30°. Do not
freeze.
Identification
A solution in dilute hydrochloric acid gives the reactions of
aluminium salts (2.3.1).
Dried Aluminium Hydroxide Gel
Tests
Dried Aluminium Hydroxide; Hydrated Aluminium Oxide
pH (2.4.24). 5.5 to 8.0.
Dried Aluminium Hydroxide Gel consists largely of hydrated
Arsenic (2.3.10). Dissolve 10.0 g in 18 ml of brominated aluminium oxide together with varying quantities of basic
hydrochloric acid, add 42 ml of water and remove the excess aluminium carbonate and bicarbonate.
bromine with a few drops of stannous chloride solution AsT.
The resulting solution complies with the limit test for arsenic Dried Aluminium Hydroxide Gel contains not less than
(1 ppm). 47.0 per cent and not more than 60.0 per cent of Al2O3.
Heavy metals (2.3.13). Dissolve 5.0 g in 10 ml of dilute Description. A white, light, amorphous powder containing
hydrochloric acid, filter if necessary, and dilute to 25 ml with some aggregates; odourless; tasteless.
80
IP 2007 ALUMINIUM SULPHATE
Identification Storage. Store protected from moisture.

A solution in dilute hydrochloric acid gives the reactions of
aluminium salts (2.3.1).
Tests Aluminium Sulphate

pH (2.4.24). Not more than 10.0, determined in a 4.0 per cent Al2(SO4)3,xH2O Mol. Wt. 342.1 (anhydrous)
w/v suspension in carbon dioxide-free water. Aluminium Sulphate contains not less than 51.0 per cent and
Arsenic (2.3.10). Dissolve 2 g in 18 ml of brominated not more than 59.0 per cent of Al2(SO4)3. It contains a variable
hydrochloric acid, add 42 ml of water and remove the excess quantity of water of crystallisation.
of bromine with a few drops of stannous chloride solution Description. Colourless, lustrous crystals or a white,
AsT. The resulting solution complies with the limit test for crystalline powder or masses; odourless.
arsenic (5 ppm).
Heavy metals (2.3.13). Dissolve 0.33 g in 10 ml of dilute Identification
hydrochloric acid with the aid of heat, filter if necessary, and
A. Dissolve 2.5 g in sufficient water to produce 50 ml (solution
dilute to 25 ml with water. The resulting solution complies
A). The solution gives reaction A of aluminium salts (2.3.1).
with the limit test for heavy metals Method A (60 ppm).
B. Solution A gives reaction A of sulphates (2.3.1).
Chlorides (2.3.12). Dissolve 0.1 g in 10 ml of dilute nitric acid,
boil, cool, dilute to 100 ml with water and filter. 20 ml of the
Tests
filtrate complies with the limit test for chlorides (1.25 per cent).
Sulphates (2.3.17). Dissolve 0.5 g in 5 ml of dilute hydrochloric Appearance of solution. Solution A is not more opalescent
acid, boil, cool, dilute to 200 ml with water and filter. 5 ml of than opalescence standard OS3 (2.4.1), and is colourless
the filtrate complies with the limit test for sulphates (1.2 per (2.4.1).
cent). pH (2.4.24). 2.5 to 4.0, determined in a 2.0 per cent w/v solution.
Neutralising capacity. Pass a sufficient quantity, triturated if Alkalis and alkaline-earth metals. To 20 ml of solution A add
necessary, through a sieve of nominal mesh aperture of 100 ml of water, heat and add 0.1 ml of methyl red solution.
150 µm. Weigh accurately 0.5 g of the sifted material and add Add 6 M ammonia until the colour changes to yellow, dilute
to 200.0 ml of 0.05 M hydrochloric acid previously heated to to 150 ml with water, heat to boiling and filter. Evaporate 75 ml
37° and stir continuously, maintaining the temperature at 37°; of the filtrate to dryness on a water-bath and ignite. The weight
the pH of the solution, at 37°, after 10, 15 and 20 minutes, is of the residue does not exceed 2 mg (0.4 per cent).
not less than 1.8, 2.3 and 3.0 respectively and at no time is
more than 4.5. Add 10.0 ml of 0.5 M hydrochloric acid Ammonium salts. Heat 1 g with 10 ml of sodium hydroxide
previously heated to 37°, stir continuously for 1 hour solution on a water-bath for 1 minute; the odour of ammonia
maintaining the temperature at 37° and titrate with 0.1 M sodium is not perceptible.
hydroxide to pH 3.5. Arsenic (2.3.10). Dissolve 3.3 g in 50 ml of water and add 10 ml
Not more than 35.0 ml of 0.1 M sodium hydroxide is required of stannated hydrochloric acid AsT. The resulting solution
and the pH of the solution at 37° at no time is more than 4.5. complies with the limit test for arsenic (3 ppm).
Microbial contamination (2.2.9). 1 g is free from Escherichia Heavy metals (2.3.13). 0.5 g dissolved in 1 ml of dilute acetic
coli. acid and sufficient water to produce 25 ml complies with the
limit test for heavy metals, Method A (40 ppm).
Assay. Weigh accurately about 0.4 g and dissolve in a mixture
of 3 ml of hydrochloric acid and 3 ml of water by warming on Iron (2.3.14). 8 ml of solution A diluted to 10 ml complies with
a water-bath, cool to below 20° and dilute to 100.0 ml with the limit test for iron, but using 0.3 ml in place of 0.1 ml of
water. To 20.0 ml of this solution, add 40.0 ml of 0.05 M thioglycollic acid (100 ppm).
disodium edetate, 80 ml of water, and 0.15 ml of methyl red Assay. Weigh accurately about 0.6 g and dissolve in 2 ml of
solution and neutralise by the dropwise addition of 1 M sodium 1 M hydrochloric acid and 50 ml of water. Add 50.0 ml of
hydroxide. Warm on a water-bath for 30 minutes, add 3 g of 0.05 M disodium edetate and neutralise to methyl red solution
hexamine and titrate with 0.05 M lead nitrate using 0.5 ml of with 1 M sodium hydroxide. Heat the solution to boiling,
xylenol orange solution as indicator. leave on a water-bath for 10 minutes, cool rapidly and add
1 ml of 0.05 M disodium edetate is equivalent to 0.002549 g of about 50 mg of xylenol orange mixture and 5 g of hexamine.
Al2O3. Titrate with 0.05 M lead nitrate. Carry out a blank titration.
81
AMANTADINE HYDROCHLORIDE IP 2007
1 ml of 0.05 M disodium edetate is equivalent to 0.008554 g of Related substances. Determine by gas chromatography
Al2 (SO4)3. (2.4.13).
examination in 2 ml of water, add 2 ml of a 20 per cent w/v
Amantadine Hydrochloride solution of sodium hydroxide and 2 ml of chloroform and
shake for 10 minutes. Separate the chloroform layer, dry over
NH2 anhydrous sodium sulphate and filter.
– a glass column 1.8 m x 2 mm, packed with material
, HCl
prepared in the following manner. Mix 19.5 g of silanised
diatomaceous support (such as Chromosorb G/AW/
DMCS) with 60 ml of a 0.33 per cent w/v solution of
C10H17N,HCl Mol. Wt. 187.7
potassium hydroxide in methanol and evaporate the
3,7
Amantadine Hydrochloride is tricyclo[3.3.1.1 ]dec-1- solvent under reduced pressure while slowly rotating
ylamine hydrochloride. the mixture. Dissolve over a 5-hour period 0.4 g of low-
Amantadine Hydrochloride contains not less then 98.5 per vapour pressure hydrocarbons (type L) (such as
cent and not more than 101.0 per cent of C10H17N,HCl, calculated Apiezon L) in 60 ml of toluene, add this solution to the
on the anhydrous basis. prepared silanised diatomaceous support and evaporate
the solvent under reduced pressure while slowly rotating
Description. A white or almost white, crystalline powder; the mixture,
sublimes when heated. – temperature:
column. Allow the temperature to increase from 100° to
Identification 200° at a constant rate of 6° per minute,
Test A may be omitted if tests B and C are carried out. Test B inlet port. 220°,
may be omitted if tests A and C are carried out. detector. 300 °,
– flow rate. 30 ml per minute of the carrier gas.
A. Dissolve 0.1 g in 5 ml of water, add 0.5 ml of 5 M sodium
Inject 1 µl or other suitable volume of the test solution. Record
hydroxide, extract with 5 ml of dichloromethane, filter the
the chromatogram for at least 2.5 times the retention time of
dichloromethane layer through anhydrous sodium sulphate
the principal peak.
with 2 ml of dichloromethane and evaporate the solution to
dryness. The residue complies with the following test. The area of any secondary peak is not greater than 0.3 per
cent and the sum of the areas of any secondary peaks is not
greater than 1 per cent by normalisation.
Compare the spectrum with that obtained with amantadine
hydrochloride RS treated in the same manner or with the Sulphated ash (2.3.18). Not more than 0.1 per cent.
reference spectrum of amantadine. Water (2.3.43). Not more than 0.5 per cent, determined on
B. Dissolve 0.2 g in 1 ml of 0.1 M hydrochloric acid and add 2.0 g.
1 ml of a 50 per cent w/v solution of sodium nitrite; a white Assay. Weigh accurately about 0.15 g, dissolve in a mixture of
precipitate is produced. 5.0 ml of 0.01 M hydrochloric acid and 50 ml of ethanol
C. 1 ml of a 10 per cent w/v solution in carbon dioxide-free (95 per cent) and titrate with 0.1 M sodium hydroxide
water gives reaction A of chlorides (2.3.1). determining the end-point potentiometrically (2.4.25). Record
the volume used between the two inflections.
Tests
1 ml of 0.1 M sodium hydroxide is equivalent to 0.01877 g of
Appearance of solution. A 10.0 per cent w/v solution in carbon C10H17N,HCl.
dioxide-free water is clear (2.4.1) and not more intensely
coloured than reference solution YS7 (2.4.1).
Amantadine Capsules
Heavy metals (2.3.13). A solution prepared by dissolving 1.0 g Amantadine Hydrochloride Capsules
in 1 ml of dilute acetic acid and sufficient water to produce Amantadine Capsules contain not less than 95.0 per cent and
25 ml complies with the limit test for heavy metals, Method A not more than 105.0 per cent of the stated amount of
(20 ppm). amantadine hydrochloride, C10H17N,HCl.
82
IP 2007 AMBROXOL HYDROCHLORIDE
Identification hydrochloride RS or with the reference spectrum of ambroxol

hydrochloride.
To the contents of three capsules add 6 ml of pentane and
shake well. Collect the undissolved solids on a sintered-glass B. Dissolve 25 mg in 2.5 ml of water, add 1.0 ml of dilute
filter, wash with two portions, each of 3 ml, of pentane and dry ammonia and allow to stand for 5 minutes. Acidify the aqueous
in air. The residue comply with the following tests. layer with dilute nitric acid and filter. The filtrate gives reaction
A of chlorides (2.3.1).
Compare the spectrum with that obtained with amantadine Tests
hydrochloride RS treated in the same manner or with the
pH (2.4.24). 4.5 to 6.0, determined in a 1.0 per cent w/v solution
reference spectrum of amantadine.
in carbon dioxide-free water.
B. Dissolve 0.2 g in 1 ml of 0.1 M hydrochloric acid and add
1 ml of a 50 per cent w/v solution of sodium nitrite; a white
(2.4.14).
precipitate is produced.
Tests examination in water and dilute to 50 ml with the same solvent.
Other tests. Comply with the tests stated under Capsules. Reference solution (a). Dissolve 5 mg of ambroxol
hydrochloride RS in 250 ml of water. Dilute 5 ml of the solution
Assay. Weigh accurately a quantity of the mixed contents of
to 100 ml with the mobile phase.
20 capsules containing about 0.12 g of Amantadine
Hydrochloride and warm in a mixture of 30 ml of anhydrous Reference solution (b). Dissolve 5 mg of the substance under
glacial acetic acid and 10 ml of mercuric acetate solution. examination in 0.2 ml of methanol and add 0.04 ml of a mixture
Titrate with 0.1 M perchloric acid, using crystal violet of 1 volume of formaldehyde solution and 99 volumes of water.
solution as indicator. Carry out a blank titration. Heat at 60º for 5 minutes. Evaporate to dryness under a current
of nitrogen. Dissolve the residue in 5 ml of water and dilute to
1 ml of 0.1 M perchloric acid is equivalent to 0.01877 g of 20 ml with the mobile phase.
C10H17N,HCl.
Storage. Store protected from moisture at a temperature not – a stainless steel column 25 cm x 4 mm, packed with
exceeding 30°. octadecylsilane bonded to porous silica (5 µm),
– mobile phase: a mixture of equal volumes of acetonitrile
and a buffer solution prepared by dissolving 1.32 g of
Ambroxol Hydrochloride ammonium phosphate in 900 ml of water, adjusting the
pH to 7.0 with phosphoric acid and diluting to 1000 ml
C13H18Br2N2O,HCl Mol. Wt. 414.6 with water,
OH – spectrophotometer set at 248 nm,
Br Inject reference solution (b). The test is not valid unless the
N
H , HCl resolution between the secondary peak (trans-4-(6,8-dibromo-
NH2 1,4-dihydroquinazolin-3(2H)-yl)cyclohexanol) and the
Br ambroxol peak is at least 4.0.
Inject the test solution and reference solution (a). Continue
Ambroxol hydrochloride is trans-4-[(2-amino-3,5- the chromatography for 3 times the retention time of the
dibromobenzyl)amino]cyclohexanol hydrochloride. principal peak in the chromatogram obtained with the test
Ambroxol Hydrochloride contains not less than 99.0 per cent solution. The area of any secondary peak in the chromatogram
and not more than 101.0 per cent of C13H18Br2N2O,HCl, obtained with the test solution is not more than 5 times the
calculated on the dried basis. area of the principal peak in the chromatogram obtained with
reference solution (a) (0.5 per cent). The sum of the areas of all
Description. A white or yellowish crystalline powder.
the secondary peaks is not more than 10 times the area of the
Identification principal peak in the chromatogram obtained with reference
solution (a) (1.0 per cent). Ignore any peak with an area 0.1
A. Determine by infrared absorption spectrophotometry (2.4.6). times the area of the principal peak in the chromatogram
Compare the spectrum with that obtained with ambroxol obtained with reference solution (a) (0.01 per cent).
83
AMIKACIN IP 2007
Heavy metals (2.3.13). 1.0 g complies with the limit test for Apply to the plate 3 µl of each solution. After development,
heavy metals, Method B (20 ppm). allow the plate to dry in air, heat it at 110° for 15 minutes and
Sulphated ash (2.3.18). Not more than 0.1 per cent. immediately spray it with a 1 per cent w/v solution of ninhydrin
in a mixture of 100 volumes of 1-butanol and 1 volume of
Loss on drying (2.4.19). Not more than 0.5 per cent, determined pyridine. The principal pink-coloured spot in the
on 1.0 g by drying in an oven at 105º. chromatogram obtained with the test solution corresponds to
Assay. Dissolve 0.3 g in 70 ml of ethanol. Titrate with 0.1 M those in the chromatograms obtained with reference solutions
sodium hydroxide, determining the end-point (a) and (b).
potentiometrically (2.4.25). Carry out a blank titration. B. To 1 ml of a 1 per cent w/v solution add 1 ml of 2 M sodium
1 ml of 0.1 M sodium hydroxide is equivalent to 0.04146 g of hydroxide, mix and add 2 ml of a 1 per cent w/v solution of
C13H18Br2N2O, HCl. cobalt nitrate; a violet colour is produced.
Storage. Store protected from light. C. To a solution of 50 mg in 5 ml of water add 4 ml of a 0.035 per
cent w/v solution of anthrone in sulphuric acid; a bluish-
violet colour is produced.
Amikacin Tests
HO in carbon dioxide-free water.
O Specific optical rotation (2.4.22). +97 ° to +105°, determined in
O a 2.0 per cent w/v solution.
NH2
OH NH2 Sulphated ash (2.3.18). Not more than 1.0 per cent, the charred
O HN
H2N residue being moistened with 2 ml of nitric acid and 5 drops
OH OH of sulphuric acid.
O HO
OH 0.2 g.
O NH2
OH
Assay. Determine by the microbiological assay of antibiotics,
OH Method B (2.2.10), and express the result in µg of Amikacin,
C22H43N5O13, per mg.
C22H43N5O13 Mol. Wt. 585.6
Amikacin is (S)-O-3-amino-3-deoxy-α-D-glucopyranosyl-
(1→ 6)-O-[6-amino-6-deoxy-α-D-glucopyranosyl(1→ 4)]- N1- Amikacin Sulphate
(4-amino-2-hydroxy-1-oxobutyl)-2-deoxy-D-streptamine.
HO
Amikacin contains not less than 900 µg of C22H43N5O13 per
mg, calculated on the anhydrous basis. O
O
Description. A white crystalline powder; almost odourless. NH2
OH NH2 , H2SO4
O HN
Identification H2N OH OH
O HO
A. Determine by thin-layer chromatography (2.4.17), coating
the plate with silica gel G. OH
O NH2
OH
Mobile phase. A mixture of 60 volumes of methanol, 30 volumes
of strong ammonia solution and 25 volumes of chloroform. OH
Test solution. A 0.6 per cent w/v solution of the substance C22H43N5O13,1.8H2SO4 Mol. Wt.762.1
under examination. C22H43N5O13,2H2SO4 Mol. Wt. 781.8
Reference solution (a). A 0.6 per cent w/v solution of Amikacin Sulphate is (S)-O-3-amino-3-deoxy-α-D-
amikacin RS. glucopyranosyl-(1→ 6)-O-[6-amino-6-deoxy-α-D-
Reference solution (b). A mixture of equal volumes of the test glucopyranosyl(1→4)]-N1-(4-amino-2-hydroxy-1-oxobutyl)-
solution and reference solution (a). 2-deoxy-D-streptamine sulphate (1:2 or 1:1.8)(salt).
84
IP 2007 AMIKACIN INJECTION
Amikacin Sulphate having a molar ratio of Amikacin to H2SO4 Assay. Determine by the microbiological assay of antibiotics,
of 1:2 contains the equivalent of not less than 674 µg and not Method B (2.2.10), and express the result in µg of amikacin,
more than 786 µg of C22H43N5O13 per mg, calculated on the C22H43N5O13, per mg.
dried basis. Amikacin Sulphate having a molar ratio of Amikacin Labelling. The label states (1) whether the molar ratio of
to H2SO4 of 1:1.8 contains the equivalent of not less than amikacin to H2SO4 of the contents is 1:2 or 1:1.8; (2) whether
691µg and not more than 806 µg of C22H43N5O13 per mg, the material is intended for use in the manufacture of parenteral
calculated on the dried basis. preparations.
Description. A white to yellowish-white crystalline powder;
almost odourless.
Amikacin Injection
Identification
Amikacin Sulphate Injection
the plate with silica gel G. Amikacin Injection is a sterile solution of Amikacin Sulphate
in Water for Injections or of Amikacin in Water for Injections
Mobile phase. A mixture of 60 volumes of methanol, 30 volumes prepared with the aid of Sulphuric Acid.
of strong ammonia solution and 25 volumes of chloroform.
Amikacin Injection contains not less than 90.0 per cent and
Test solution. A 0.6 per cent w/v solution of the substance not more than 120.0 per cent of the stated amount of amikacin,
under examination. C22H43N5O13.
amikacin RS. Identification
Reference solution (b). A mixture of equal volumes of the test Dilute the injection to obtain a solution containing 6 mg of
solution and reference solution (a). amikacin per ml (test solution). The test solution complies
with the following tests.
allow the plate to dry in air, heat it at 110° for 15 minutes and A. Determine by thin-layer chromatography (2.4.17), coating
immediately spray it with a 1 per cent w/v solution of ninhydrin the plate with silica gel G.
in a mixture of 100 volumes of 1-butanol and 1 volume of Mobile phase. A mixture of 60 volumes of methanol,
pyridine. The principal pink-coloured spot in the 30 volumes of strong ammonia solution and 25 volumes of
chromatogram obtained with the test solution corresponds to chloroform.
those in the chromatograms obtained with reference solutions
(a) and (b). Reference solution (a). A 0.6 per cent w/v solution of
amikacin RS.
B. To 1 ml of a 1 per cent w/v solution add 1 ml of 2 M sodium
Reference solution (b). A mixture of equal volumes of the test
hydroxide, mix and add 2 ml of a 1 per cent w/v solution of
solution and reference solution (a).
cobalt nitrate; a violet colour is produced.
C. To a solution of 50 mg in 5 ml of water add 4 ml of a 0.035 per
allow the plate to dry in air, heat it at 110° for 15 minutes and
cent w/v solution of anthrone in sulphuric acid; a bluish-
immediately spray it with a 1 per cent w/v solution of ninhydrin
violet colour is produced.
in a mixture of 100 volumes of 1-butanol and 1 volume of
Tests pyridine. The principal pink-coloured spot in the
chromatogram obtained with the test solution corresponds to
pH (2.4.24). 2.0 to 4.0 (1:2 salt), or 6.0 to 7.3 (1:1.8 salt), those in the chromatograms obtained with reference solutions
determined in a 1.0 per cent w/v solution in carbon dioxide- (a) and (b).
free water.
B. To 1.5 ml of the test solution add 1 ml of 2 M sodium
Specific optical rotation (2.4.22). +76.0° to +84.0°, determined hydroxide, mix and add 2 ml of a 1 per cent w/v solution of
in a 2.0 per cent w/v solution. cobalt nitrate; a violet colour is produced.
Sulphated ash (2.3.18). Not more than 1.0 per cent, the charred C. To 1.5 ml of the test solution add 3.5 ml of water mix and add
residue being moistened with 2 ml of nitric acid and 5 drops 4 ml of a 0.035 per cent w/v solution of anthrone in sulphuric
of sulphuric acid. acid; a bluish-violet colour is produced.
Tests
on 0.1 g by drying in an oven over phosphorus pentoxide at
110° at a pressure not exceeding 0.7 kPa for 3 hours. pH (2.4.24). 3.5 to 5.5.
85
AMILORIDE HYDROCHLORIDE IP 2007
Bacterial Endotoxins (2.2.3). Not more than 0.33 Endotoxin examine in ultraviolet light at 365 nm. The principal spot in the
unit per mg of amikacin. chromatogram obtained with the test solution corresponds to
Other tests. Complies with the tests stated under Parenteral that in the chromatogram obtained with the reference solution.
Preparations (Injections). C. Dissolve 10 mg in 10 ml of water and add 10 ml of a 20 per
cent w/v solution of cetrimide, 0.25 ml of 2 M sodium hydroxide
Assay. Dilute the injection to obtain a solution containing
and 1 ml of bromine water; a greenish-yellow colour is
1 mg of amikacin per ml. Determine by the microbiological
produced. Add 2 ml of 2 M hydrochloric acid; the solution
assay of antibiotics, Method B, (2.2.10) and express the result
becomes deep yellow and exhibits a blue fluorescence in
in mg of amikacin, C22H43N5O13 per ml.
ultraviolet light at 365 nm.
Labelling. The label states the quantity of Amikacin Sulphate
D. A 5 per cent w/v solution gives the reactions of chlorides
contained in the sealed container in terms of the equivalent
(2.3.1).
amount of amikacin.
Tests
Free acid. Dissolve 1.0 g in 100 ml of a mixture of equal volumes
Amiloride Hydrochloride of methanol and water and titrate with 0.1 M sodium
hydroxide determining the end-point potentiometrically
O NH (2.4.25); not more than 0.3 ml is required.
Cl N Related substances. Determine by liquid chromatography
N NH2
, HCl , 2H2O (2.4.14).
H
H2N N NH2 Solvent mixture. A mixture of 1 volume of acetonitrile and 3
volumes of water.
C6H8ClN7O,HCl,2H2O Mol. Wt. 302.1
Test solution (a) Dissolve 0.2 g of the substance under
Amiloride Hydrochloride is N-amidino-3,5-diamino-6- examination in 100 ml of solvent mixture.
chloropyrazine-2-carboxamide hydrochloride dihydrate.
Test solution (b). Dilute 1 ml of test solution (a) to 100 ml with
Amiloride Hydrochloride contains not less than 98.0 per cent the same solvent mixture.
and not more than 101.0 per cent of C6H8ClN7O,HCl, calculated
Test solution (c). Dilute 10 ml of test solution (b) to 100 ml
with the same solvent mixture.
Description. A pale yellow to greenish-yellow powder.
Reference solution. A 0.001 per cent w/v solution of methyl
Identification 3,5-diamino- 6-chloropyrazine-2-carboxylate RS in the same
solvent mixture.
Test A may be omitted if tests B, C and D are carried out. Tests Chromatographic system
B and C may be omitted if tests A and D are carried out. – a stainless steel column 25 cm x 4.6 mm, packed with
A. Determine by infrared absorption spectrphotometry (2.4.6). octadecylsilane chemically bonded to porous silica or
Compare the spectrum with that obtained with amiloride ceramic microparticles (5 µm) (such as Nucleosil C18),
hydrochloride RS or with the reference spectrum of amiloride – mobile phase: a mixture of 745 volumes of water,
hydrochloride. 250 volumes of acetonitrile and 5 volumes of
tetramethylammonium hydroxide solution (10 per cent),
B. Determine by thin-layer chromatography (2.4.17), coating
the pH of the mixture being adjusted to 7.0 with a mixture
the plate with a suitable silica gel.
of 1 volume of phosphoric acid and 9 volumes of water,
Mobile phase. A freshly prepared mixture of 88 volumes of – flow rate. 1 ml per minute,
dioxan, 6 volumes of dilute ammonia solution and 6 volumes – spectrophotometer set at 254 nm,
of water. – a 20 µl loop injector.
Test solution. Dissolve 0.2 g of the substance under Inject the reference solution and adjust the concentration of
examination in sufficient methanol to produce 50 ml. acetonitrile so that the retention time of methyl 3,5-diamino-
6-chloropyrazine-2-carboxylate is 5 to 6 minutes (an increase
Reference solution. A 0.4 per cent w/v solution of amiloride
in the concentration of acetonitrile reduces the retention time).
hydrochloride RS in methanol.
Inject 20 µl of test solution (b) and adjust the concentrations
Apply to the plate 5 µl of each solution. Allow the mobile of tetramethylammonium hydroxide and orthophosphoric
phase to rise 12 cm. Dry the plate in a current of warm air and acid so that the retention time of amiloride is 9 to 12 minutes
86
IP 2007 AMILORIDE TABLETS
keeping the pH at 7.0 (an increase in the concentrations reduces The principal spot in the chromatogram obtained with the test
the retention time). solution corresponds to that in the chromatogram obtained
Inject each of test solution (a) and the reference solution and with the reference solution.
allow the chromatography to proceed for 5 times the retention
Tests
time of amiloride. In the chromatogram obtained with test
solution (a) the sum of the areas of any secondary peaks is Related substances. Determine by thin-layer chromatography
not greater than the area of the peak due to methyl 3,5-diamino- (2.4.17), using a precoated silica gel plate (such as Merck
6-chloropyrazine-2-carboxylate in the chromatogram obtained silica gel 60 plates).
with the reference solution. Ignore any peak with an area less
than 10 per cent of the area of the peak due to methyl 3,5- Mobile phase. A freshly prepared mixture of 90 volumes of
diamino-6- chloropyrazine-2-carboxylate in the chromatogram dioxan and 12 volumes of 3 M ammonia.
obtained with the reference solution. Test solution. Shake a quantity of the powdered tablets
Inject test solution (c). The test is not valid if the signal-to- containing 17.5 mg of anhydrous amiloride hydrochloride with
noise ratio of the peak due to amiloride in the chromatogram 5 ml of methanol and centrifuge.
obtained with this solution is less than 5.0. Reference solution (a). A 0.002 per cent w/v solution of methyl
Sulphated ash (2.3.18). Not more than 0.1 per cent. 3,5-diamino-6-chloropyrazine-2-carboxylate RS in
methanol.
Water (2.3.43). 11.0 to 13.0 per cent, determined on 0.2 g.
Reference solution (b). A 0.0008 per cent w/v solution of methyl
Assay. Weigh accurately about 0.25 g and dissolve in a mixture 3,5-diamino-6-chloropyrazine-2- carboxylate RS in
of 100 ml of anhydrous glacial acetic acid and 15 ml of dioxan methanol.
and add 10 ml of mercuric acetate solution Titrate with 0.1 M
perchloric acid, determining the end-point potentiometrically Apply to the plate 5 µl of each solution. After development,
(2.4.25). Carry out a blank titration. dry the plate in air and examine in ultraviolet light at 365 nm.
Any spot corresponding to methyl 3,5-diamino-6-
1 ml of 0.1 M perchloric acid is equivalent to 0.02661 g of chloropyrazine-2-carboxylate in the chromatogram obtained
C6H8ClN7O,HCl. with the test solution is not more intense than the spot in
Storage. Store protected from light. chromatogram obtained with reference solution (a). Any other
secondary spot in the chromatogram obtained with the test
solution is not more intense than the spot in the chromatogram
obtained with reference solution (a) and not more than one
Amiloride Tablets such spot is more intense than the spot in the chromatogram
obtained with reference solution (b).
Amiloride Hydrochloride Tablets
Uniformity of content. Comply with the test stated under
Amiloride Tablets contain not less than 90.0 per cent and not Tablets.
more than 110.0 per cent of the stated amount of anhydrous
amiloride hydrochloride, C6H8ClN7O,HCl. Powder one tablet and transfer to a 100-ml volumetric flask,
add 60 ml of 0.1 M hydrochloric acid, and shake by
Identification mechanical means for 30 minutes. Dilute with 0.1 M
hydrochloric acid to volume, mix, and centrifuge a portion of
A. Extract a quantity of the powdered tablets containing the mixture. Dilute an accurately measured portion of the clear
0.5 mg of anhydrous amiloride hydrochloride with 100 ml of supernatant liquid quantitatively to obtain a solution
0.1 M hydrochloric acid and filter. When examined in the containing about 10 µg of amiloride hydrochloride per ml.
range 230 nm to 380 nm (2.4.7), the solution shows absorption Measure the absorbance of the resulting solution at the
maxima at about 285 nm and at about 363 nm. maximum at about 363 nm (2.4.7).
B. Carry out the method described under Related substances Calculate the content of C6H8ClN7O, HCl taking 692 as the
using the following solutions. specific absorbance at 363 nm.
Test solution. Shake a quantity of the powdered tablets Other tests. Comply with the tests stated under Tablets.
containing 10 mg of anhydrous amiloride hydrochloride with
Assay. Weigh and finely powder 20 tablets. Weigh accurately
10 ml of methanol and centrifuge.
a quantity of the powder containing about 10 mg of anhydrous
Reference solution. A 0.1 per cent w/v solution of amiloride amiloride hydrochloride, transfer to a 100-ml volumetric flask,
hydrochloride RS in methanol. add 60 ml of 0.1 M hydrochloric acid, and shake by
87
AMINOCAPROIC ACID IP 2007
mechanical means for 30 minutes. Dilute with 0.1 M Tests

hydrochloric acid to volume, mix, and centrifuge a portion of
the mixture. Dilute an accurately measured portion of the clear Appearance of solution. A 20.0 per cent w/v solution remains
supernatant liquid quantitatively to obtain a solution clear for 24 hours (2.4.1), and is colourless (2.4.1).
containing about 10 µg of amiloride hydrochloride per ml. pH (2.4.24). 7.5 to 8.0, determined in a 20.0 per cent w/v solution.
Measure the absorbance of the resulting solution at the Stability. Place 20.0 g evenly spread in a shallow dish about 9
maximum at about 363 nm (2.4.7). cm in diameter, cover and allow to stand at 100° ± 2° for
Calculate the content of C6H8ClN7O,HCl taking 692 as the 72 hours. Dissolve in sufficient water to produce 100.0 ml.
specific absorbance at 363 nm. Prepare a 20.0 per cent w/v solution of the substance under
examination but without the above treatment. Measure the
absorbances (2.4.7) of the two solutions at the maximum at
Labelling. The label states the strength in terms of the about 287 nm and at about 450 nm. Absorbance of the solution
equivalent amount of anhydrous amiloride hydrochloride. prepared from the exposed substance being examined at the
maximum at about 287 nm is not more than 0.15 and of the
solution of the substance under examination without the above
treatment, at the maximum at about 287 nm is not more than
Aminocaproic Acid 0.10. Absorbance of both solutions at the maximum at about
450 nm is not more than 0.03.
O
H2 N heavy metals, Method B (10 ppm).
OH
C6H13NO2 Mol. Wt. 131.2
Aminocaproic Acid is 6-aminohexanoic acid. on 1.0 g by drying in an oven at 105°.
Aminocaproic Acid contains not less than 98.5 per cent and Assay. Weigh accurately about 0.2 g, dissolve in about 100 ml
not more than 101.0 per cent of C6H13NO2, calculated on the of anhydrous glacial acetic acid with gentle heat to effect
dried basis. solution, cool and add 15 ml of mercuric acetate solution.
Titrate with 0.1 M perchloric acid, using crystal violet
Description. Colourless crystals or a white, crystalline powder.
solution as indicator. Carry out a blank titration.
Identification 1 ml of 0.1 M perchloric acid is equivalent to 0.01312 g of
C6H13NO2.
Compare the spectrum with that obtained with aminocaproic
acid RS. Aminocaproic Acid Injection
B. Determine by thin-layer chromatography (2.4.7), coating Aminocaproic Acid Injection is a sterile solution of
the plate with silica gel G. Aminocaproic Acid in Water for Injections.
Mobile phase. A mixture of 25 volumes of ethanol (95 per Aminocaproic Acid Injection contains not less than 92.5 per
cent), 3 volumes of water and 4 volumes of strong ammonia cent and not more than 107.5 per cent of the stated amount of
solution. aminocaproic acid, C6H13NO2.
examination in 100 ml of water. Identification
Reference solution. A 0.25 per cent w/v solution of To a volume containing 0.4 g of Aminocaproic Acid add 2 ml
aminocaproic acid RS. of ether, stir, add 2 ml of methanol, stir again and allow to
stand; the crystals after drying on a water-bath comply with
Apply to the plate 2 µl of each solution. After development, the following tests.
remove the plate, spray it with a 0.25 per cent w/v solution of
ninhydrin in a mixture of equal volumes of methanol and A. Determine by infrared absorption spectrophotometry (2.4.6).
pyridine and heat at 105° for 2 minutes. The principal spot in Compare the spectrum with that obtained with aminocaproic
the chromatogram obtained with the test solution corresponds acid RS.
to that in the chromatogram obtained with the reference B. Determine by thin-layer chromatography (2.4.17), coating
solution. the plate with silica gel G.
88
IP 2007 AMINOPHYLLINE
Mobile phase. A mixture of 25 volumes of ethanol (95 per B. Determine by thin-layer chromatography (2.4.7), coating
cent), 3 volumes of water and 4 volumes of strong ammonia the plate with silica gel G.
solution. Mobile phase. A mixture of 25 volumes of ethanol (95 per
Test solution. Dissolve 0.25 g of the substance under cent), 3 volumes of water and 4 volumes of strong ammonia
examination in 100 ml of water. solution.
Reference solution. A 0.25 per cent w/v solution of Test solution. Dissolve 0.25 g of the substance under
aminocaproic acid RS. examination in 100 ml of water.
Apply to the plate 2 µl of each solution. After development, Reference solution. A 0.25 per cent w/v solution of
remove the plate, spray it with a 0.25 per cent w/v solution of aminocaproic acid RS.
ninhydrin in a mixture of equal volumes of methanol and Apply to the plate 2 µl of each solution. After development,
pyridine and heat at 105° for 2 minutes. The principal spot in remove the plate, spray it with a 0.25 per cent w/v solution of
the chromatogram obtained with the test solution corresponds ninhydrin in a mixture of equal volumes of methanol and
to that in the chromatogram obtained with the reference pyridine and heat at 105° for 2 minutes. The principal spot in
solution. the chromatogram obtained with the test solution corresponds
Tests to that in the chromatogram obtained with the reference
solution.
pH (2.4.24). 6.0 to 7.6.
Tests
Bacterial endotoxins (2.2.3). Not more than 0.05 Endotoxin
Unit per mg of aminocaproic acid. Other tests. Comply with the tests stated under Tablets.
Other tests. Complies with the tests stated under Parenteral Assay. Weigh and powder 20 tablets. Weigh accurately a
Preparations (Injections). quantity of the powder containing about 0.2 g of Aminocaproic
Acid, add about 100 ml of anhydrous glacial acetic acid,
Assay. To a volume containing 0.2 g of Aminocaproic Acid
heat gently to effect solution, cool and add 15 ml of mercuric
add 10 ml of ethanol and evaporate to dryness on a water-
acetate solution. Titrate with 0.1 M perchloric acid, using
bath. Dissolve the residue in 100 ml of anhydrous glacial
acetic acid by gentle heating, if necessary, cool and add 15 ml
of mercuric acetate solution. Titrate with 0.1 M perchloric 1ml of 0.1 M perchloric acid is equivalent to 0.01312 g of
acid, using crystal violet solution as indicator. Carry out a C6H13NO2.
blank titration.
C6H13NO2. Aminophylline
Theophylline and Ethylenediamine
Aminocaproic Acid Tablets O

H
Aminocaproic Acid Tablets contain not less than 95.0 per H3C N NH2
N
cent and not more than 105.0 per cent of the stated amount of
aminocaproic acid, C6H13NO2. O N N
NH2
CH3
Identification 2
Triturate 2 tablets with 10 ml of water and filter into 100 ml of

(C7H8N4O2)2,C2H8N2 Mol. Wt. 420.4 (anhydrous)
acetone. Swirl the mixture and allow to stand for 15 minutes to
complete crystallisation. Filter through a medium porosity, Aminophylline is a stable mixture or combination of
sintered-glass filter and wash the crystals with 25 ml of acetone. theophylline and ethylenediamine. It may be anhydrous or
Apply vacuum to remove the solvent, dry at 105° for 30 minutes may contain not more than two molecules of water of
and cool. The residue complies with the following tests. hydration.
A. Determine by infrared absorption spectrophotometry (2.4.6). Aminophylline contains the equivalent of not less than
Compare the spectrum with that obtained with aminocaproic 84.0 per cent and not more than 87.4 per cent of theophylline,
acid RS. C7H8N4O2, and the equivalent of not less than 13.5 per cent
89
AMINOPHYLLINE INJECTION IP 2007
and not more than 15.0 per cent of ethylenediamine, C2H8N2, Apply to the plate 10 µl of each solution. After development,
both calculated on the anhydrous basis. dry the plate in air and examine in ultraviolet light at 254 nm.
Description. A white or slightly yellowish granules or powder; Any secondary spot in the chromatogram obtained with the
odour, slightly ammoniacal. On exposure to air it gradually test solution is not more intense than the spot in the
loses ethylenediamine and absorbs carbon dioxide with chromatogram obtained with the reference solution.
liberation of free theophylline. Even in the absence of light, it Heavy metals (2.3.13). A 8 per cent w/v solution complies with
is gradually decomposed on exposure to a humid environment, the limit test for heavy metals, Method A (20 ppm).
the degradation being faster at higher temperatures.
Identification Water (2.3.43). Not more than 1.5 per cent (for anhydrous),
Test A may be omitted if tests B, C and D are carried out. Tests determined on 2.0 g dissolved in 20 ml of pyridine. 3.0 to 8.0
B, C and D may be omitted if test A is carried out. per cent (for hydrate), determined on 0.5 g.
Dissolve 1 g in 10 ml of water and add 2 ml of dilute Assay. For theophylline — Weigh accurately about 0.25 g,
hydrochloric acid dropwise, with shaking. Separate the add 50 ml of water and 8 ml of dilute ammonia solution and
precipitate by filtration and reserve the filtrate for test D. Wash warm gently on a water-bath until complete solution is effected.
the precipitate with successive small quantities of cold water, Add 20.0 ml of 0.1 M silver nitrate, mix and boil for 15 minutes.
recrystallise from hot water and dry at 100° to 105°.The residue Cool to between 5° and 10° for 20 minutes, filter at a pressure
complies with the following tests. not exceeding 2.75 kPa and wash the precipitate with three
quantities, each of 10 ml, of water. Acidify the combined filtrate
and washings with nitric acid and add an excess of 3 ml of the
Compare the spectrum with that obtained with theophylline
acid. Cool, add 2 ml of ferric ammonium sulphate solution,
RS.
and titrate with 0.1 M ammonium thiocyanate.
B. To 10 mg of the residue obtained in test A add 1 ml of
hydrochloric acid in a porcelain dish and 0.1 g of potassium 1 ml of 0.1 M silver nitrate is equivalent to 0.01802 g of
chlorate and evaporate to dryness on a water-bath; invert the C7H8N4O2.
dish over a vessel containing a few drops of dilute ammonia For ethylenediamine — Weigh accurately about 0.25 g and
solution; the residue acquires a purple colour. Add a few drops dissolve in 30 ml of water. Titrate with 0.1 M hydrochloric
of dilute sodium hydroxide solution; the colour is discharged. acid using methyl orange solution as indicator.
C. Saturate in water a portion of the residue obtained in test A 1 ml of 0.1 M hydrochloric acid is equivalent to 0.003005 g of
and add tannic acid solution; a precipitate soluble in excess C2H8N2.
of the reagent is produced.
Storage. Store protected from light and from atmospheric
D. The filtrate complies with the following test. carbon dioxide.
To the filtrate reserved above add 0.2 ml of benzoyl chloride,
make alkaline with 2 M sodium hydroxide and shake vigorously.
Filter, wash the precipitate with 10 ml of water, dissolve in 5 ml
of hot ethanol (95 per cent) and add 5 ml of water. The
precipitate, after washing with water and drying at 100° to
Aminophylline Injection
105° melts at 248° to 252° (2.4.21). Theophylline and Ethylenediamine Injection
Tests Aminophylline Injection is a sterile solution of Aminophylline
in Water for Injections or is a sterile solution of Theophylline
Related substances. Determine by thin-layer chromatography in a solution of Ethylenediamine Hydrate in Water for
(2.4.17) coating the plate with silica gel GF254. Injections free from carbon dioxide. Aminophylline Injection
Mobile phase. A mixture of 40 volumes of 1-butanol, 30 may contain an excess of ethylenediamine but no other
volumes of acetone, 30 volumes of chloroform and 10 volumes substance may be added.
of strong ammonia solution.
Aminophylline Injection contains theophylline, C7H8N4O2,
Test solution. Dissolve 0.2 g of the substance under equivalent to not less than 73.25 per cent and not more than
examination in 2 ml of water with the aid of heat and dilute to 88.25 per cent of the stated amount of aminophylline, and not
10 ml with methanol. more than 0.295 g of ethylenediamine, C2H8N2 for each g of
Reference solution. Dilute 1 volume of the test solution to 200 anhydrous theophylline, C7H8N4O2, determined in the Assay
volumes with methanol. for theophylline.
90
IP 2007 AMINOPHYLLINE TABLETS
Identification Calculate the amount of C2H8N2 present for each g of C7H8N4O2

found.
Dilute a volume containing about 0.5 g of aminophylline with
water to about 25 ml and add 1 ml of dilute hydrochloric acid Storage. Store in single dose containers, from which carbon
with constant stirring. Separate the precipitate by filtration dioxide has been excluded. Do not allow contact with metals.
and reserve the filtrate for test D. Wash the precipitate with a Labelling. The label states (1) the strength in terms of the
small portion of cold water, recrystallise from hot water and equivalent amount of anhydrous aminophylline in a suitable
dry at 100° to 105°. The crystalline powder complies with the dose-volume; (2) the route of injection; (3) that the injection is
following tests. not to be used if crystals have separated.
Compare the spectrum with that obtained with theophylline
RS. Aminophylline Tablets
B. To 10 mg add 1 ml of hydrochloric acid in a porcelain dish Theophylline and Ethylenediamine Tablets
and 0.1 g of potassium chlorate and evaporate to dryness on
Aminophylline Tablets contain theophylline, C7H8N4O2,
a water-bath; invert the dish over a vessel containing a few
equivalent to not less than 80.6 per cent and not more than
drops of dilute ammonia solution; the residue acquires a
90.8 per cent of the stated amount of aminophylline, and
purple colour. Add a few drops of dilute sodium hydroxide
ethylenediamine, C2H8N2, equivalent to not less than 10.9 per
solution; the colour is discharged.
cent of the stated amount of aminophylline.
C. Saturate a portion in water and add tannic acid solution; a
precipitate soluble in excess of the reagent is produced. Identification
The filtrate complies with the following test. Shake a quantity of the powdered tablets containing 0.5 g of
Add 0.2 ml of benzoyl chloride, make alkaline with 2 M sodium aminophylline with 25 ml of water and filter. To the filtrate add
hydroxide and shake vigorously. Filter, wash the precipitate 1 ml of dilute hydrochloric acid with constant stirring. Separate
with 10 ml of water, dissolve in 5 ml of hot ethanol (95 per the precipitate by filtration and reserve the filtrate. Wash the
cent) and add 5 ml of water. The precipitate, after washing precipitate with a small portion of cold water, recrystallise
with water and drying at 100° to 105° melts at 248° to 252° from hot water and dry at 100° to 105°. The crystalline powder
(2.4.21). complies with the following tests.
Tests Compare the spectrum with that obtained with theophylline
pH (2.4.24). 8.8 to 10.0. RS.
Other tests. Complies with the tests stated under Parenteral B. To 10 mg add 1 ml of hydrochloric acid in a porcelain dish
Preparations (Injections). and 0.1 g of potassium chlorate and evaporate to dryness on
Assay. For theophylline — Measure accurately a volume a water-bath; invert the dish over a vessel containing a few
containing about 0.25 g of aminophylline and dilute with water drops of dilute ammonia solution; the residue acquires a
to 40 ml. Add 8 ml of dilute ammonia solution and 20.0 ml of purple colour. Add a few drops of dilute sodium hydroxide
0.1 M silver nitrate, mix and boil for 15 minutes. Cool to solution; the colour is discharged.
between 5° and 10° for 20 minutes, filter at a pressure not C. Saturate a portion in water and add tannic acid solution; a
exceeding 2.75 kPa and wash the precipitate with three precipitate soluble in excess of the reagent is produced.
quantities, each of 10 ml, of water. Acidify the combined filtrate The filtrate complies with the following test.
and washings with nitric acid and add an excess of 3 ml of the
acid. Cool, add 2 ml of ferric ammonium sulphate solution, Add 0.2 ml of benzoyl chloride, make alkaline with 2 M sodium
and titrate with 0.1 M ammonium thiocyanate. hydroxide and shake vigorously. Filter, wash the precipitate
with 10 ml of water, dissolve in 5 ml of hot ethanol (95 per
1 ml of 0.1 M silver nitrate is equivalent to 0.01802 g of cent) and add 5 ml of water. The precipitate, after washing
C7H8N4O2. with water and drying at 100° to 105° melts at 248° to 252°
For ethylenediamine — To a volume containing about 0.25 (2.4.21).
g of aminophylline, add sufficient water to produce 30 ml.
Tests
Titrate with 0.1 M hydrochloric acid using methyl orange
solution as indicator. Other tests. Comply with the tests stated under Tablets.
1 ml of 0.1 M hydrochloric acid is equivalent to 0.003005 g of Assay. For theophylline — Weigh and powder 20 tablets.
C2H8N2. Weigh accurately a quantity of the powdered tablets
91
AMIDARONE HYDROCHLORIDE IP 2007
containing about 0.5 g of aminophylline, transfer to a 200-ml C. Gives reaction A of chlorides (2.3.1).
volumetric flask with the aid of a mixture of 50 ml of water and
15 ml of dilute ammonia solution and allow to stand for Tests
30 minutes with frequent shaking, warming to about 50°, if
necessary. Cool, add water to volume and mix. Centrifuge the Appearance of solution. A 5.0 per cent w/v solution is clear
mixture, and pipette a volume of the clear supernatant liquid (2.4.1), and not more intensely coloured than reference solution
equivalent to about 0.25 g of aminophylline into a flask, dilute GY5 (2.4.1).
with sufficient water to produce 40 ml and add 8 ml of dilute pH (2.4.24). 3.2 to 3.8, determined in 5.0 per cent w/v solution,
ammonia solution. Add 20.0 ml of 0.1 M silver nitrate, mix prepared by dissolving in carbon dioxide-free water at 80°
and boil for 15 minutes. Cool to between 5° and 10° for 20 and cooling.
minutes, filter at a pressure not exceeding 2.75 kPa and wash
Related substances. Determine by thin layer chromatography
the precipitate with three quantities, each of 10 ml, of water.
Acidify the combined filtrate and washings with nitric acid
and add an excess of 3 ml of the acid. Cool, add 2 ml of ferric All the solutions should be protected from light and should
ammonium sulphate solution, and titrate with 0.1 M be used immediately after preparation.
ammonium thiocyanate. Mobile phase. A mixture of 5 volumes of anhydrous formic
1 ml of 0.1 M silver nitrate is equivalent to 0.01802 g of acid, 10 volumes of methanol and 85 volumes of
C7H8N4O2. dichloromethane.
Storage. Store protected from light. Test solution (a). Dissolve 10 g of the substance under
examination in 100 ml of dichloromethane.
Labelling. The label states the strength in terms of the
equivalent amount anhydrous aminophylline. Test solution (b). Dissolve 0.5 g of the substance under
Amiodarone Hydrochloride amiodarone hydrochloride RS in dichloromethane.
Reference solution (b). A 0.05 per cent w/v solution of the
H 3C O substance under examination in dichloromethane.
I CH3 Reference solution (c). A 0.025 per cent w/v solution of the
O , HCl substance under examination in dichloromethane.
N CH3
O
Reference solution (d). A 0.02 per cent w/v of (2-chloroethyl)
I diethylamine hydrochloride RS in dichloromethane.
C25H29I2NO3,HCl Mol. Wt. 681.8 Apply to the plate 5 µl of each solution. After development,
dry in a current of cold air and examine in ultraviolet light at
Amiodarone Hydrochloride is 2-butylbenzofuran-3yl-4-(2- 254 nm. Any secondary spot in the chromatogram obtained
diethylaminoethoxy)-3,5-diiodophenyl ketone hydrochloride. with test solution (a) is not more intense than the spot in the
Amiodarone Hydrochloride contains not less than 98.5 per chromatogram obtained with reference solution (c) (0.05 per
cent and not more than 101.0 per cent of C25H29I2NO3,HCl, cent) and not more than one such spot is more intense than
calculated on the dried basis. the spot in the chromatogram obtained with reference solution
(d) (0.25 per cent). Spray with potassium iodobismuthate
Description. A white or almost white, fine crystalline powder.
solution. Examine immediately in daylight. Any spot
Identification corresponding to (2-chloroethyl) diethylamine hydrochloride
in the chromatogram obtained with test solution (a) is not
Text B may be omitted if tests A and C are carried out. Test A more intense than the spot in the chromatogram obtained
may be omitted if tests B and C are carried out. with reference solution (d) (0.2 per cent).
A. Determine by infrared absorption spectrophotometry (2.4.6). Iodides. Dissolve 1.5 g in 40 ml of water at 80° by shaking until
Compare the spectrum with that obtained with amiodarone completely dissolved. Cool and dilute to 50 ml with water
hydrochloride RS. (Solution A).
B. In the test for Related substances the principal spot in the To 15 ml of solution A add 1 ml of 0.1 M hydrochloric acid
chromatogram obtained with test solution (b) corresponds to and 1ml of 0.05 M potassium iodate and dilute to 20 ml with
that in the chromatogram obtained with reference solution (a). water. Allow to stand protected from light for 4 hours (Solution
92
IP 2007 AMIODARONE TABLETS
1). To 15 ml of solution A add 1 ml of 0.1 M hydrochloric acid, Tests

1 ml of an 88.2 ppm solution of potassium iodide and 1 ml of
0.05 M potassium iodate and dilute to 20 ml with water. Allow
(2.4.17), coating the plate with silica gel GF254
to stand protected from light for 4 hours (Solution 2). Measure
the absorbances of solutions (1) and (2) at the maximum at Mobile phase. A mixture of 85 volumes of dichloromethane,
about 420 nm, using as the blank a mixture of 15 ml of solution 10 volumes of methanol and 5 volumes of anhydrous formic
A and 1 ml of 0.1 M hydrochloric acid diluted to 20 ml with acid.
water (2.4.7). The absorbance of solution (1) is not greater Test solution. Shake a quantity of the powdered tablets
than half the absorbance of solution (2) (150 ppm). containing 50 mg of Amiodarone Hydrochloride with 20 ml of
Heavy metals (2.3.13). 10 g complies with the limit test for methanol and filter.
heavy metals, Method C (20 ppm). Reference solution (a). Dilute 1 volume of the test solution to
Sulphated ash (2.3.18). Not more than 0.1 per cent. 200 volumes with methanol.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Reference solution (b). A 0.00125 per cent w/v solution of
on 1.0 g by drying at 100° at a pressure not exceeding 0.3 kPa 2-butyl-3-(4-hydroxy-3, 5-diiodobenzoyl) benzofuran RS in
for 4 hours. methanol.
Apply separately to the plate (pre-washed with the mobile
phase and dried in air before use) 10 µl of each solution After
of 5.0 ml of 0.01 M hydrochloric acid and 75 ml of ethanol
development, dry the plate in a current of warm air and examine
(95 per cent). Titrate with 0.1 M sodium hydroxide,
in ultraviolet light at 254 nm. In the chromatogram obtained
determining the end-point potentiometrically (2.4.25). Carry
with the test solution any spot corresponding to 2-butyl-3 (4-
out a blank titration.
hydroxy-3, 5-diiodobenzoyl)benzofuran is not more intense
1 ml of 0.1 M sodium hydroxide is equivalent to 0.06818 g of than the spot in the chromatogram obtained with reference
C25H29I2NO3, HCI. solution (b) (0.5 per cent) and any other secondary spot is not
Storage. Store protected from light, at a temperature not more intense than the spot in the chromatogram obtained
exceeding 30°. with reference solution (a) (0.5 per cent).
Assay. Weigh and powder 20 tablets.
Amiodarone Tablets Determine by liquid chromatography (2.4.14).
Amiodarone Hydrochloride Tablets Test solution. Weigh accurately a quantity of the powdered
tablets containing about 0.1g of Amiodarone Hydrochloride,
Amiodarone Tablets contain not less than 95.0 per cent and
add 70 ml of methanol, mix with the aid of ultrasound for
15 minutes, cool and dilute to 100.0 ml with the same solvent
amiodarone hydrochloride, C25H29I2NO3, HCI.
and filter. Dilute 10.0 ml of the filtrate to 100.0 ml with the
Identification mobile phase.
A. Shake a quantity of the powdered tablets containing about Reference solution. Dissolve 0.1g of amiodarone
0.3 g of Amiodarone Hydrochloride with 25 ml of hydrochloride RS in 70 ml of methanol, cool and dilute to
dichloromethane, filter and evaporate the filtrate to dryness. 100.0 ml with the same solvent. Dilute 10.0 ml of the resulting
To the residue, add 2 ml of 1 M sodium hydroxide and extract solution to 100.0 ml with the mobile phase.
with 25 ml of ether. Dry the extract over anhydrous sodium Chromatographic system
sulphate, filter and evaporate to dryness. Dry the residue – a stainless steel column 7.5 cm x 3.9 mm, packed with
obtained under reduced pressure over phosphorus pentoxide very finely divided silica gel consisting of porous
and dissolve in 2.5 ml of dichloromethane. The solution spherical particles with chemically bonded nitrile group
complies with the following test. (4 µm), (such as Nova-Pack CNHP),
– mobile phase: a mixture of 45 volumes of 0.01 M sodium
perchlorate and 55 volumes of acetonitrile, the pH of
Compare the spectrum with that obtained with amiodarone
the mixture being adjusted to 3.0 with 2 M phosphoric
hydrochloride RS.
acid,
B. In the Assay, the principal peak in the chromatogram – flow rate. 1 ml per minute,
obtained with the test solution corresponds to the peak in the – spectrophotometer set at 244 nm,
chromatogram obtained with the reference solution. – 20 µl loop injector.
93
AMITRIPTINE HYDROCHLORIDE IP 2007
Inject the reference solution. The relative standard deviation pH (2.4.24). 4.5 to 6.0, determined in a 1.0 per cent w/v solution.
for replicate injections is not more than 2.0 per cent. Heavy metals (2.3.13). 1.0 g complies with the limit test for
Inject the test solution and the reference solution. heavy metals, Method B (20 ppm).
Calculate the content of C25H29I2NO3, HCI in the tablets. Related substances. Determine by thin-layer chromatography
(2.4.17), protected from light, coating the plate with silica gel
G.
Amitriptyline Hydrochloride Mobile phase. A mixture of 85 volumes of cyclohexane,

15 volumes of ethyl acetate and 3 volumes of diethylamine.
Test solution. Weigh accurately about 0.2 g of the substance
under examination and dissolve in sufficient chloroform to
produce 10 ml.
CH3 Reference solution (a). A 0.001 per cent w/v solution of
N , HCl
dibenzosuberone RS in chloroform.
CH3
cyclobenzaprine hydrochloride RS in chloroform.
C20H23N,HCl Mol. Wt. 313.9 Apply to the plate 10 µl of each solution. Allow the mobile
phase to rise 14 cm in an unlined tank. Dry the plate in air until
Amitriptyline Hydrochloride is 3-(10,11-dihydro-5H- the odour of the solvent is no longer detectable, spray with a
dibenzo[a,d]cyclohept-5-ylidene)propyldimethylamine freshly prepared mixture of 4 volumes of formaldehyde
hydrochloride. solution and 96 volumes of sulphuric acid, heat at 105° for
Amitriptyline Hydrochloride contains not less than 99.0 per 10 minutes and examine in ultraviolet light at 365 nm. Any
cent and not more than 101.0 per cent of C20H23N, HCl, spots in the chromatogram obtained with the test solution
calculated on the dried basis. corresponding to dibenzosuberone and cyclobenzaprine
hydrochloride are not more intense than the spots in the
Description. Colourless crystals or a white or almost white
chromatograms obtained with reference solutions (a) and (b)
powder; almost odourless.
respectively and any other secondary spot is not more intense
Identification than the spot in the chromatogram obtained with reference
solution (b).
Test A may be omitted if tests B, C, and D are carried out.
Tests B and C may be omitted if tests A and D are carried out.
Compare the spectrum with that obtained with amitriptyline
hydrochloride RS or with the reference spectrum of Assay. Weigh accurately about 1.0 g and dissolve in 50 ml of
amitriptyline hydrochloride. anhydrous glacial acetic acid, warm slightly, if necessary, to
effect solution. Cool, add 10 ml of mercuric acetate solution
B. When examined in the range 230 nm to 360 nm (2.4.7), a
and titrate with 0.1 M perchloric acid, using crystal violet
solution as indicator to a green end-point. Carry out a blank
maximum only at about 239 nm; absorbance at about 239 nm,
titration.
about 0.55.
C. To about 50 mg dissolved in 3 ml of water add 1 drop of a
C20H23N,HCl.
2.5 per cent w/v solution of quinhydrone in methanol; no red
colour is produced within 15 minutes (distinction from
nortriptyline).
D. Gives the reactions of chlorides (2.3.1).
Amitriptyline Tablets
Amitriptyline HydrochlorideTablets
Tests
Amitriptyline Tablets contain not less than 90.0 per cent and
Appearance of solution. Dissolve 1.25 g in sufficient water to not more than 110.0 per cent of the stated amount of
produce 25 ml. The solution is clear (2.4.1) and not more amitriptyline hydrochloride, C20H23N, HCl. The tablets are
intensely coloured than reference solution BS8 (2.4.1). coated.
94
IP 2007 AMITRIPTINE TABLETS
Identification than the spot in the chromatogram obtained with reference

solution (b).
A. Shake a quantity of the powdered tablets containing about
5 mg of Amitriptyline Hydrochloride with 20 ml of methanol Uniformity of content (For tablets containing 10 mg or less).
and filter. To 1 ml of the filtrate add 1 ml of a 2.5 per cent w/v Comply with the test stated under Tablets.
solution of sodium bicarbonate, 1 ml of a 2 per cent w/v Determine by liquid chromatography (2.4.14)
solution of sodium periodate and 1 ml of a 0.3 per cent w/v Test solution. Powder one tablet, shake with 2.5 ml of 0.1 M
solution of potassium permanganate, allow to stand for hydrochloric acid until completely disintegrated, add 5 ml of
15 minutes, acidify with dilute sulphuric acid and extract with methanol, shake for 30 minutes, dilute the suspension to
10.0 ml of 2,2,4-trimethylpentane. When examined in the 10 ml with methanol, centrifuge and use the clear supernatant
range 230 nm to 360 nm (2.4.7), the resulting solution shows liquid.
an absorption maximum only at about 265 nm.
Reference solution. Weigh accurately 25.0 mg of amitriptyline
B. Triturate a quantity of the powdered tablets containing hydrochloride RS dissolve in 10 ml of methanol and dilute to
0.1 g of Amitriptyline Hydrochloride with 10 ml of chloroform, 25.0 ml with methanol (50 per cent).
filter and evaporate the filtrate to a low volume. Add ether
until a turbidity is produced and allow to stand. To about 50
mg of the precipitate dissolved in 3 ml of water add 1 drop of
a 2.5 per cent w/v solution of quinhydrone in methanol; no
ceramic microparticles (10 µm),
red colour is produced within 15 minutes (distinction from
– mobile phase: 0.03 M sodium hexanesulphonate in a
nortriptyline).
mixture of equal volumes of acetonitrile and water,
C. The precipitate obtained in test B gives reaction A of adjusted to pH 4.5 by the addition of glacial acetic
chlorides (2.3.1). acid,
Tests – spectrophotometer set at 239 nm,
Related substances. Determine by thin-layer chromatography – a 20 µl loop injector.
(2.4.17), protected from light, coating the plate with silica gel Calculate the content of C20H23N,HCl in the tablet.
G.
Mobile phase. A mixture of 85 volumes of cyclohexane,
Apparatus. No 2
15 volumes of ethyl acetate and 3 volumes of diethylamine.
Test solution. Extract a quantity of the powdered tablets Speed and time. 100 rpm and 45 minutes.
containing 20 mg of Amitriptyline Hydrochloride with 5 ml of
a mixture of 9 volumes of ethanol (95 per cent) and 1 volume Use one tablet in the vessel for each test.
of 2 M hydrochloric acid centrifuge and use the supernatant Withdraw a suitable volume of the medium and filter. Measure
liquid, evaporated to dryness and dissolve in 10 ml of the absorbance of the filtered solution, suitably diluted with
chloroform. the medium if necessary, at the maximum at about 239 nm
(2.4.7). Calculate the content of C20H23N, HCl in the medium
from the absorbance obtained from a solution of known
dibenzosuberone RS in chloroform.
concentration of amitriptyline hydrochloride RS in the same
Reference solution (b). A 0.004 per cent w/v solution of medium.
cyclobenzaprine hydrochloride RS in chloroform.
Apply to the plate 10 µl of each solution. Allow the mobile C20H23N,HCl.
phase to rise 14 cm in an unlined tank. Dry the plate in air until Other tests. Comply with the tests stated under Tablets.
the odour of the solvent is no longer detectable, spray with a
freshly prepared mixture of 4 volumes of formaldehyde Assay. Determine by liquid chromatography (2.4.14).
solution and 96 volumes of sulphuric acid, heat at 105° for 10 Test solution. When tablets are film-coated, shake 20 tablets
minutes and examine in ultraviolet light at 365 nm. Any spots with 50 ml of 0.1 M hydrochloric acid until completely
in the chromatogram obtained with the test solution disintegrated, add 100 ml of methanol, shake for 30 minutes,
corresponding to dibenzosuberone and cyclobenzaprine dilute the suspension to 200.0 ml with methanol, centrifuge
hydrochloride are not more intense than the spots in the and dilute a volume of the supernatant liquid equivalent to
chromatograms obtained with reference solutions (a) and (b) 25 mg of Amitriptyline Hydrochloride to 100.0 ml with methanol
respectively and any other secondary spot is not more intense (50 per cent).
95
AMLODIPINE BESILATE IP 2007
When tablets are sugar-coated, weigh and powder 20 tablets. maximum at about 360 nm. The specific absorbance at the
Weigh accurately a quantity of the powder containing about maximum is 113 to 121.
50 mg of Amitriptyline Hydrochloride, shake with 50 ml of
0.1 M hydrochloric acid for 30 minutes, add 100 ml of Tests
methanol, shake for 30 minutes, dilute the mixture to 200.0 ml
with water, centrifuge and use the supernatant liquid. Optical rotation (2.4.22). - 0.10º to + 0.10º, determined in a 1.0
per cent w/v solution in methanol.
Reference solution. Dissolve 50 mg of amitriptyline
hydrochloride RS in 10 ml of methanol and dilute to 200.0 ml Related substances. A. Determine by thin-layer
with methanol (50 per cent). chromatography (2.4.17), coating the plate with silica gel
GF254.
Follow the procedure described under Uniformity of content.
Mobile phase. The upper layer of a mixture of 25 volumes of
Calculate the content of C20H23N,HCl in the tablets. glacial acetic acid, 25 volumes of water and 50 volumes of
methyl isobutyl ketone.
Amlodipine Besilate examination in 2 ml of methanol.
H methanol.
H3C N NH2
O SO3H Reference solution (a). Dissolve 70 mg of amlodipine besilate
O O CH3 RS in 1 ml of methanol.
H3C ,
O O
Cl 10 ml with methanol.
Reference solution (c). Dilute 3 ml of test solution (b) to 100
ml with methanol.
Reference solution (d). Dilute 1 ml of test solution (b) to 100
C26H31ClN2O8S Mol. Wt. 567.1 ml with methanol.
Amlodipine Besilate is 3-ethyl 5-methyl (4RS)-2-[(2-
aminoethoxy)methyl]-4-(2-chlorophenyl)-6-methyl-1,4-
phase to rise 15 cm. Dry the plate at 80º for 15 minutes and
dihydropyridine-3,5-dicarboxylate benzene sulphonate.
examine in ultraviolet light at 254 nm and 365 nm. The
Amlodipine Besilate contains not less than 97.0 per cent and chromatogram obtained with reference solution (a) shows two
not more than 102.0 per cent of C26H31ClN2O8S, calculated on clearly separated minor spots with Rf values of about 0.18 and
the anhydrous basis. 0.22. In the chromatogram obtained with reference solution
Description. A white or almost white powder. (a) any spot, other than the principal spot, is not more intense
than the spot in the chromatogram obtained with reference
Identification solution (c) (0.3 per cent) and at most 2 spots are more intense
Test A may be omitted if tests B and C are carried out. Tests B solution (d) (0.1 per cent).
and C may be omitted if test A is carried out.
B. Determine by liquid chromatography (2.4.14).
Compare the spectrum with that obtained with amlodipine Test solution (a). Dissolve 50 mg of the substance under
besilate RS or with the reference spectrum of amlodipine examination in the mobile phase and dilute to 50.0 ml with the
besilate. mobile phase.
B. In test A for Related substances, the principal spot in the Test solution (b). Dilute 5.0 ml of test solution (a) to 100.0 ml
chromatogram obtained with test solution (b) corresponds to with the mobile phase.
that in the chromatogram obtained with reference solution Reference solution (a). A solution containing 0.005 per cent
(b). w/v of amlodipine besilate RS in the mobile phase.
C. When examined in the range 300 nm to 400 nm (2.4.7), a Reference solution (b). Dilute 3 ml of test solution (a) to 100
0.005 per cent w/v solution in a 1 per cent v/v solution of 0.1 ml with the mobile phase and dilute 5 ml of the solution to 50
M hydrochloric acid in methanol shows an absorption ml with the mobile phase.
96
IP 2007 AMLODIPINE TABLETS
Reference solution (c). Dissolve 5 mg of the substance under Identification

examination in 5 ml of strong hydrogen peroxide solution.
Heat at 70º for 45 minutes. In the Assay, the principal peak in the chromatogram obtained
with the test solution corresponds to the peak in the
Chromatographic system chromatogram obtained with the reference solution.
octadecylsilane bonded to porous silica (5 µm), Tests
35 volumes of methanol and 50 volumes of a solution Related substances. Determine by liquid chromatography
prepared by dissolving 7.0 ml of triethylamine in 1000 (2.4.14).
ml of water and adjusting the pH to 3.0 with phosphoric Test solution (a). Weigh and powder 20 tablets. Weigh
acid, accurately a quantity of the powder containing about 50 mg
– flow rate. 1 ml per minute, amlodipine, dissolve in the mobile phase, dilute to 50.0 ml with
– spectrophotometer set at 237 nm, the mobile phase and centrifuge.
Test solution (b). Dilute 5.0 ml of test solution (a) to 100.0 ml
The relative retention time between amlodipine and 3-ethyl 5- with the mobile phase.
methyl(4RS)-4-(2-chlorophenyl)-6-methyl-2-[[2-[[2-
(methylcarbamoyl)benzoyl]amino]ethoxy] methyl]-1,4- Reference solution (a). A solution containing 0.005 per cent
dihydropyridine-3,5-dicarboxylate (impurity A) is about 0.5. w/v of amlodipine besilate RS in the mobile phase.
Inject reference solution (c). The test is not valid unless the Reference solution (b). Dilute 5 ml of test solution (a) to 100
resolution between the peaks corresponding to amlodipine ml with the mobile phase and dilute 5 ml of the solution to 50
and impurity A is at least 4.5. ml with the mobile phase.
Inject test solution (a) and reference solutions (b) and Reference solution (c). Dissolve 5 mg of amlodipine besilate
(c).Continue the chromatography for 3 times the retention RS in 5 ml of strong hydrogen peroxide solution. Heat at 70°
time of amlodipine. The area of any peak obtained due to for 45 minutes and centrifuge.
impurity A multiplied by 2 is not more than the area of the Chromatographic system
principal peak in the chromatogram obtained with reference – a stainless steel column 15 cm x 3.9 mm, packed with
solution (b) (0.3 per cent). The sum of the areas of all the other octadecylsilane bonded to porous silica (5 µm),
secondary peaks is not more than the area of the principal – mobile phase: a mixture of 15 volumes of acetonitrile,
peak in the chromatogram obtained with reference solution 35 volumes of methanol and 50 volumes of a solution
(b) (0.3 per cent). Ignore any peak due to benzene sulphonate prepared by dissolving 7.0 ml of triethylamine in 1000
(relative retention about 0.2) and any peak with an area 0.1 ml of water and adjust to pH 3.0 with phosphoric acid,
times the area of the principal peak in the chromatogram – flow rate. 1 ml per minute,
obtained with reference solution (b) (0.03 per cent). – spectrophotometer set at 237 nm,
Sulphated ash (2.3.18). Not more than 0.2 per cent. – a 10 µl loop injector.
Water (2.3.43). Not more than 0.5 per cent, determined on 3.0 g. The relative retention time between amlodipine and 3-ethyl 5-
Assay. Determine by liquid chromatography (2.4.14) as methyl(4RS)-4-(2-chlorophenyl)-6-methyl-2-[[2-[[2-
described under Related substances. (methylcarbamoyl)benzoyl]amino]ethoxy] methyl]-1,4-
dihydropyridine-3,5-dicarboxylate (impurity A) is about 0.5.
Inject alternately test solution (b) and reference solution (a).
Inject reference solution (c). The test is not valid unless the
Calculate the content of C26H31ClN2O8S. resolution between the peaks corresponding to amlodipine
Storage. Store protected from moisture. and impurity A is at least 4.5.
Inject test solution (a) and reference solutions (b) and (c).
Continue the chromatography for 3 times the retention time of
Amlodipine Tablets amlodipine. The area of any peak corresponding to impurity A
Amlodipine Besilate Tablets multiplied by 2 is not more than the area of the principal peak
Amlodipine Tablets contain Amlodipine Besilate.
per cent). The sum of the areas of all the other secondary
Amlodipine Tablets contain not less than 90.0 per cent and peaks is not more than the area of the principal peak in the
not more than 110.0 per cent of the stated amount of chromatogram obtained with reference solution (b) (0.5 per
amlodipine, C20H25ClN2O5. cent) Ignore any peak due to benzene sulphonate (relative
97
AMMONIUM CHLORIDE IP 2007
retention about 0.2) and any peak with an area 0.1 times the Tests
area of the principal peak in the chromatogram obtained with
reference solution (b) (0.05 per cent). Appearance of solution. A 10.0 per cent solution is clear (2.4.1)
and colourless (2.4.1).
pH (2.4.24). 4.5 to 6.0, determined in a 5.0 per cent solution.
Apparatus No. 1
Medium. 900 ml of 0.01 M hydrochloric acid. Arsenic (2.3.10) Dissolve 2.5 g in 50 ml of water and add 10 ml
of stannated hydrochloric acid. The resulting solution
Speed and time. 75 rpm and 45 minutes.
complies with the limit test for arsenic (4 ppm).
Withdraw a suitable volume of the medium and filter. Measure
the absorbance of the filtered solution, suitably diluted with
heavy metals, Method A (10 ppm).
the dissolution medium if necessary, at the maximum at about
239 nm (2.4.7). Calculate the content of C20H25ClN2O5 in the Iron (2.3.14). 2.0 g complies with the limit test for iron
medium from the absorbance obtained from a solution of known (20 ppm).
concentration of amlodipine besilate RS in the same medium. Calcium. To 0.2 ml of ethanolic calcium standard solution
D. Not less than 70 per cent of the stated amount of (100 ppm Ca) add 1 ml of a 4 per cent w/v solution of
C20H25ClN2O5. ammonium oxalate. After 1 minute add 1 ml of 2 M acetic acid
and 15 ml of a solution made by diluting 5 ml of a 10 per cent
solution of the substance under examination with 10 ml of
Tablets.
water and shake. Compare any opalescence produced with
Determine by liquid chromatography (2.4.14) as described that of a standard prepared in a similar manner but using a
under Related substances using the following solutions. mixture of 10 ml of calcium standard solution (10 ppm Ca)
Test solution. Powder one tablet and dissolve in 50 ml of and 5 ml of water instead of the solution of the substance
methanol, dilute with sufficient methanol to get a solution under examination (200 ppm).
containing 0.002 per cent w/v of amlodipine, shake for 10 Sulphates (2.3.17). 1.0 g complies with the limit test for
minutes and filter through a glass-fibre filter paper. sulphates (150 ppm).
Reference solution. A solution of amlodipine besilate RS in Thiocyanate. Acidify 10 ml of a 10 per cent w/v solution with
methanol equivalent to 0.002 per cent w/v of amlodipine. hydrochloric acid and add a few drops of ferric chloride
Calculate the content of C20H25ClN2O5 in the tablet. solution; no red colour is produced.
Other tests. Comply with the tests stated under Tablets. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Assay. Determine by liquid chromatography (2.4.14) as Loss on drying (2.4.19). Not more than 1.0 per cent determined
described under Related substances. on 1.0 g by drying in an oven at 105°.
Inject alternately test solution (b) and reference solution (a). Assay. Weigh accurately about 0.1 g, dissolve in 20 ml of
water and add a mixture of 5 ml of formaldehyde solution,
Calculate the content of C20H25ClN2O5 in the tablets. previously neutralised to dilute phenolphthalein solution,
Storage. Store protected from moisture. and 20 ml of water. After 2 minutes, titrate slowly with 0.1 M
sodium hydroxide using a further 0.2 ml of dilute
phenolphthalein solution as indicator.
equivalent amount of amlodipine.
NH4Cl.
Ammonium Chloride
NH4Cl Mol. Wt. 53.5 Amodiaquine Hydrochloride
Ammonium Chloride contains not less than 99.0 per cent and
not more than 100.5 per cent of NH4Cl, calculated on the dried OH CH3
N
basis. , 2HCl , 2H2O
N CH3
Description. Colourless crystals or a white, crystalline powder. N
H
Identification Cl
Gives the reactions of ammonium salts and of chlorides (2.3.1). C20H22ClN3O, 2HCl, 2H2O Mol. Wt. 464.9
98
IP 2007 AMODIAQUINE TABLETS
Amodiaquine Hydrochloride is 4-(7-chloro-4-quinolylamino)- Reference solution (b). Dilute 1 volume of reference solution
2-(diethylaminomethyl)phenol dihydrochloride dihydrate. (a) with sufficient chloroform saturated with strong ammonia
Amodiaquine Hydrochloride contains not less than 98.0 per solution to obtain 200 volumes.
cent and not more than 101.5 per cent of C20H22ClN3O, 2HCl, Apply to the plate 10 µl of each solution. Allow the mobile
calculated on the anhydrous basis. phase to rise 10 cm. Dry the plate in air and examine in ultraviolet
Description. A yellow, crystalline powder; odourless or almost
odourless.
chromatogram obtained with reference solution (a) and no
Identification secondary spot in the chromatogram obtained with the test
solution is more intense than the principal spot in the
Test A may be omitted if tests B, C, D and E are carried out. chromatogram obtained with reference solution (b).
Tests C and D may be omitted if tests A, B and E are carried Sulphated ash (2.3.18). Not more than 0.l per cent.
out.
A. Dissolve 20 mg in 10 ml of water and add 1 ml of strong
Assay. Weigh accurately about 0.3 g and dissolve in sufficient
ammonia solution. Extract with two quantities, each of 25 ml,
0.1 M hydrochloric acid to produce 200.0 ml. Dilute 10.0 ml to
of chloroform, wash the combined chloroform extracts with
1000.0 ml with 0.1 M hydrochloric acid. Measure the
water, dry with anhydrous sodium sulphate, evaporate the
absorbance of the resulting solution at the maximum at about
chloroform and dry the residue at 105° for 2 hours. The residue
343 nm (2.4.7), using 0.1 M hydrochloric acid as the blank.
complies with the following test.
Calculate the content of C 20H 22ClN3O, 2HCl from the
Determine by infrared absorption spectrophotometry (2.4.6). absorbance obtained by carrying out the Assay simultaneously
Compare the spectrum with that obtained with amodiaquine on amodiaquine hydrochloride RS.
RS.
0.0015 per cent w/v solution in 0.1 M hydrochloric acid shows
an absorption maximum at about 343 nm; absorbance at 343
Amodiaquine Tablets
nm, about 0.55. Amodiaquine Hydrochloride Tablets
C. To 1 ml of a 2 per cent w/v solution add 0.5 ml of cobalt Amodiaquine Tablets contain not less than 95.0 per cent and
thiocyanate solution; a green precipitate is produced. not more than 105.0 per cent of the stated amount of
D. To 20 ml of a 2 per cent w/v solution, add 1 ml of dilute amodiaquine, C20H22ClN3O.
ammonia solution. Shake and filter; the filtrate gives the
Identification
reactions of chlorides (2.3.1).
E. The undried material melts at about 158° (2.4.21). A. Extract the powdered tablets with water and filter. To 1 ml
of the filtrate add 0.5 ml of cobalt thiocyanate solution; a
Tests green precipitate is produced.
pH (2.4.24). 3.6 to 4.6, determined in a 2.0 per cent w/v solution. B. The powdered tablets give the reactions of chlorides (2.3.1).
Related substances. Determine by thin-layer chromatography Tests

Mobile phase. A mixture of 90 volumes of chloroform saturated (2.4.17), coating the plate with silica gel GF254.
with strong ammonia solution and 10 volumes of ethanol. Mobile phase. A mixture of 90 volumes of chloroform saturated
Test solution. Add to 200 mg of the substance under with strong ammonia solution and 10 volumes of ethanol.
examination in a glass-stoppered test-tube 10 ml of chloroform
saturated with strong ammonia solution, shake vigorously
containing 40 mg of Amodiaquine Hydrochloride with 20 ml of
for 2 minutes, allow the solids to settle and decant the
water for 1 minute, add 25 ml of chloroform and 1 ml of strong
supernatant liquid.
ammonia solution and shake vigorously for 2 minutes. Filter
Reference solution (a). Prepare in the same manner as the test the chloroform extract through a cotton plug previously soaked
solution but using 200 mg of amodiaquine hydrochloride RS in chloroform, evaporate the filtrate to dryness and dissolve
and 10 ml of chloroform saturated with strong ammonia the residue in 2 ml of chloroform saturated with strong
solution. ammonia solution.
99
AMOXYCILLIN SOIDUM IP 2007
Reference solution (a). Prepare in the same manner as the test Amoxycillin Sodium is sodium (6R)-6-(α -D-4-
solution but using 200 mg of amodiaquine hydrochloride RS hydroxyphenylglycylamino)penicillanate.
and 10 ml of chloroform saturated with strong ammonia Amoxycillin Sodium contains not less than 85.0 per cent and
solution not more than 100.5 per cent of C16H18N3NaO5S, calculated on
Reference solution (b). Dilute 1 volume of reference solution the anhydrous basis.
(a) with sufficient chloroform saturated with strong ammonia Description. A white or almost white powder; very
solution to obtain 200 volumes. hygroscopic.
phase to rise 10 cm. Dry the plate in air and examine in ultraviolet Identification
light at 254 nm. The principal spot in the chromatogram A. Determine by infrared absorption spectrophotometry (2.4.6).
obtained with the test solution corresponds to that in the Compare the spectrum with that obtained with amoxycillin
chromatogram obtained with reference solution (a) and no sodium RS or with the reference spectrum of amoxycillin
secondary spot in the chromatogram obtained with the test sodium.
solution is more intense than the principal spot in the
chromatogram obtained with reference solution (b). B. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak in the
Other tests. Comply with the tests stated under Tablets. chromatogram obtained with the reference solution.
Assay. Weigh and powder 20 tablets. Weigh accurately a C. A 5 per cent w/v solution gives the reactions of sodium
quantity of the powder containing about 0.3 g of amodiaquine, salts (2.3.1).
add 100 ml of 0.1 M hydrochloric acid and heat on a water-
bath for about 15 minutes with occasional stirring. Cool, transfer Tests
to a 200-ml graduated flask and dilute to volume with 0.1 M
hydrochloric acid. To 10.0 ml of the clear supernatant liquid
more opalescent than opalescence standard OS2 (2.4.1) when
in a separator, add 10 ml of 0.1 M hydrochloric acid and
examined immediately after preparation. The solution may
extract with 20 ml of chloroform. Discard the chloroform extract.
initially show a pink colour and its absorbance after 5 minutes
Add 4.5 ml of 1 M sodium hydroxide and extract with four
at about 430 nm is not more than 0.20 (2.4.7).
quantities, each of 25 ml, of chloroform. Extract the combined
chloroform solutions with three quantities, each of 50 ml, of pH (2.4.24). 8.0 to 10.0, determined in a 10.0 per cent w/v
0.1 M hydrochloric acid and dilute with sufficient 0.1 M solution.
hydrochloric acid to produce 200.0 ml. Dilute 10.0 ml with Specific optical rotation (2.4.22). +240° to +290°, determined
sufficient 0.1 M hydrochloric acid to produce 100.0 ml. in a 0.25 per cent w/v solution in a 0.4 per cent w/v solution of
Measure the absorbance of the resulting solution at the potassium hydrogen phthalate.
maximum at about 343 nm (2.4.7), using 0.1 M hydrochloric
acid as the blank.
heavy metals, Method B (20 ppm).
Calculate the content of C 20H 22ClN3O, 2HCl from the
N,N-Dimethylaniline (2.3.21). Not more than 20 ppm,
absorbance obtained by carrying out the Assay simultaneously
determined by Method A.
on amodiaquine hydrochloride RS. Multiply the result by
0.830 to get the equivalent quantity of C20H22ClN3O. Sodium chloride. Not more than 2.0 per cent, calculated on
the anhydrous basis, determined by the following method.
Weigh accurately about 1.0 g, dissolve in 50 ml of distilled
equivalent amount of amodiaquine.
water; add 10 ml of 2 M nitric acid and titrate with 0.1 M
silver nitrate, determining the end-point potentiometrically
(2.4.25) using a silver indicator electrode and a mercury-
Amoxycillin Sodium mercurous sulphate reference electrode or any other suitable
electrode.
H COONa
O 1 ml of 0.1 M silver nitrate is equivalent to 0.005845 g of NaCl.
NH2 N CH3
H 2-Ethylhexanoic acid. Not more than 2.0 per cent, determined
N S CH3 by the following method.
H H
O Determine by gas chromatography (2.4.13).
HO
Test solution. Prepare a 1.0 per cent w/v solution of valeric
C16H18N3NaO5S Mol. Wt. 387.4 acid (internal standard) in hexane (solution A). Dissolve 1.0 g
100
IP 2007 AMOXYCILLIN CAPSULES
of the substance under examination in 5 ml of water in a glass- Inject alternately the test solution and the reference solution.
stoppered flask, add 3 ml of 2 M hydrochloric acid, 1 ml of Calculate the percentage content of C16H18N3NaO5S by
solution A and 5 ml of hexane, shake vigorously for 1 minute, multiplying the percentage content of C16H19N3O5S by 1.060.
centrifuge if necessary and use the clear supernatant layer.
Amoxycillin Sodium intended for use in the manufacture of
Reference solution (a). Prepare in the same manner as the test parenteral preparations without a further appropriate
solution but using an extra 1 ml of hexane in place of solution procedure for the removal of bacterial endotoxins complies
A. with the following additional requirement.
Reference solution (b). Prepare in the same manner as the test Bacterial endotoxins (2.2.3). Not more than 0.25 Endotoxin
solution but using 20 mg of 2-ethylhexanoic acid suspended Unit per mg of amoxycillin.
in 5 ml of water in place of the substance under examination.
Amoxycillin Sodium intended for use in the manufacture of
Chromatographic system parenteral preparations without a further appropriate
– a glass column 1.8 m x 4 mm, packed with a support sterilisation procedure complies with the following
impregnated with a stationary phase suitable for the additional requirement.
separation of free fatty acids (such as a column Sterility (2.2.11). Complies with the test for sterility.
containing 10 per cent of SP 1200 and 1 per cent of
phosphoric acid on Chromosorb W AW, 80-100 mesh), Storage. Store protected from moisture, at a temperature not
– temperature: exceeding 30°. If it is intended for use in the manufacture of
column. 145°, parenteral preparations, the container should be sterile, tamper-
– inlet port and detector. 150°, evident and sealed so as to exclude micro-organisms.
– flow rate. 45 ml per minute of the carrier gas. Labelling. The label states whether or not the material is
Water (2.3.43). Not more than 4.0 per cent, determined on intended for use in the manufacture of parenteral preparations.
0.4 g.
Assay. Determine by liquid chromatography (2.4.14). Amoxycillin Capsules
Solvent mixture. Dissolve 6.8 g of monobasic potassium Amoxycillin Trihydrate Capsules; Amoxicillin Trihydrate
phosphate in 100 ml of water and adjust the pH to about 5.0
Capsules; Amoxicillin Capsules
with a 4.5 per cent w/v solution of potassium hydroxide.
Amoxycillin Capsules contain not less than 90.0 per cent and
Test solution. Transfer an accurately weighed quantity
containing about 120 mg of Amoxycillin to a 100-ml volumetric
amoxycillin, C16H19N3O5S.
flask, dissolve in the solvent mixture and dilute to 100.0 ml
with the solvent mixture. Use this solution within 6 hours. Identification
Reference solution. Weigh accurately a suitable quantity of Shake a quantity of the contents of the capsules containing
amoxycillin trihydrate RS, dissolve in the solvent mixture by 0.5 g of amoxycillin with 5 ml of water for 5 minutes, filter,
shaking and mixing if necessary, with the aid of ultrasound wash the residue first with ethanol and then with ether and
and dilute to obtain a solution having a known concentration dry at a pressure not exceeding 0.7 kPa for 1 hour. The residue
of about 1.2 mg per ml. Use this solution within 6 hours. complies with the following tests.
Chromatographic system Test A may be omitted if test B is carried out.
Compare the spectrum with that obtained with amoxycillin
trihydrate RS or with the reference spectrum of amoxycillin
– mobile phase: a mixture of 96 volumes of acetonitrile
trihydrate.
and 4 volumes of the solvent mixture,
– flow rate. 1.5 ml per minute, B. In the Assay, the principal peak in the chromatogram
– spectrophotometer set at 230 nm, obtained with the test solution corresponds to the peak in the
– a 10 µl loop injector. chromatogram obtained with the reference solution.
Inject the reference solution. The test is not valid unless the Tests
capacity factor is between 1.1 and 2.8, the column efficiency is
not less than 1700 theoretical plates, the tailing factor is not Dissolution (2.5.2).
more than 2.5 and the relative standard deviation for replicate Apparatus. No 1
injections is not more than 2.0 per cent. Medium. 900 ml of water.
101
AMOXYCILLIN INJECTION IP 2007
Speed and time. 100 rpm and 60 minutes. Amoxycillin Injection

Use one capsule in the vessel for each test.
Amoxicillin Sodium Injection; Amoxycillin Sodium
Withdraw a suitable volume of the medium and filter promptly Injection
through a membrane filter disc having an average pore diameter
not greater than 1.0 µm, rejecting the first 1 ml of the filtrate. Amoxycillin Injection is a sterile material consisting of
Dilute the filtrate, if necessary, with the same solvent. Measure Amoxycillin Sodium with or without excipients. It is filled in a
the absorbance of the resulting solution at the maximum at sealed container.
about 272 nm (2.4.7). Similarly measure the absorbance of a The injection is constituted by dissolving the contents of the
standard solution of known concentration of amoxicillin sealed container in the requisite amount of sterile Water for
tryhydrate RS at about 272 nm and calculate the content of Injections, immediately before use.
C16H19N3O5S.
D. Not less than 80 per cent of the stated amount of Clarity of solution and Particulate matter stated under
C16H19N3O5S. Parenteral Preparations (Injections).
Other tests. Comply with the tests stated under Capsules. Storage. The constituted solution should be used immediately
Assay. Determine by liquid chromatography (2.4.14). after preparation but, in any case, within the period
Solvent mixture. Dissolve 6.8 g of monobasic potassium
phosphate in 100 ml of water and adjust the pH to about Amoxycillin Injection contains not less than 90.0 per cent
5.0 with a 4.5 per cent w/v solution of potassium hydroxide. and not more than 120.0 per cent of the stated amount of
Test solution. Weigh accurately a quantity of the mixed amoxycillin, C16H19N3O5S.
contents of 20 capsules containing about 100 mg of amoxicillin, Description. A white or almost white powder; very
add about 80 ml of the solvent mixture and dissolve by shaking hygroscopic.
for 15 minutes and mixing if necessary, with the aid of
The contents of the sealed container comply with the
ultrasound. Dilute to 100.0 ml with the solvent mixture and
requirements stated under Parenteral Preparations
filter. Use this solution within 6 hours.
(Powders for Injection) and with the following requirements.
Reference solution. Weigh accurately a suitable quantity of
amoxycillin trihydrate RS, dissolve in the solvent mixture by Identification
shaking and mixing if necessary, with the aid of ultrasound
and dilute to obtain a solution having a known concentration A. Determine by infrared absorption spectrophotometry (2.4.6).
of about 1.2 mg per ml. Use this solution within 6 hours. Compare the spectrum with that obtained with amoxycillin
sodium RS or with the reference spectrum of amoxycillin
Chromatographic system sodium.
octadecylsilane bonded to porous silica particles or B. In the Assay, the principal peak in the chromatogram
ceramic microparticles (5 µm), obtained with the test solution corresponds to the peak in the
– mobile phase: a mixture of 96 volumes of acetonitrile chromatogram obtained with the reference solution.
and 4 volumes of the solvent mixture, C. A 5 per cent w/v solution gives the reactions of sodium
– flow rate. 1.5 ml per minute, salts (2.3.1).
– a 10 µl loop injector. Tests
more opalescent than opalescence standard OS2 (2.4.1) when
not less than 1700 theoretical plates, the tailing factor is not
examined immediately after preparation. The solution may
more than 2.5 and the relative standard deviation for replicate
initially show a pink colour and its absorbance after 5 minutes
injections is not more than 2.0 per cent.
at about 430 nm is not more than 0.20 (2.4.7).
pH (2.4.24). 8.0 to 10.0, determined in a 10.0 per cent w/v
Calculate the content of C16H19N3O5S in the capsules. solution.
Storage. Store protected from moisture. Specific optical rotation (2.4.22). +240° to +290°, determined
Labelling. The label states the quantity of the active ingredient in a 0.25 per cent w/v solution in a 0.4 per cent w/v solution of
in terms of the equivalent amount of amoxycillin. potassium hydrogen phthalate.
102
IP 2007 AMOXYCILLIN ORAL SUSPENSION
Heavy metals (2.3.13). 1.0 g complies with the limit test for Calculate the percentage content of C16H19N3O5S in the
heavy metals, Method A (20 ppm). injection.
N,N-Dimethylaniline (2.3.21). Not more than 20 ppm, Storage. Store protected from moisture, in a sterile, tamper-
determined by Method A. evident container sealed so as to exclude micro-organisms, at
a temperature not exceeding 30°.
Sodium chloride. Not more than 2.0 per cent, calculated on
the anhydrous basis, determined by the following method. Labelling. The label states the quantity of Amoxycillin Sodium
Weigh accurately about 1.0 g, dissolve in 50 ml of distilled contained in the sealed container in terms of the equivalent
water; add 10 ml of 2 M nitric acid and titrate with 0.1 M amount of amoxycillin.
silver nitrate, determining the end-point potentiometrically
(2.4.25) using a silver indicator electrode and a mercury-
mercurous sulphate reference electrode or any other suitable
electrode. Amoxycillin Oral Suspension
1 ml of 0.1 M silver nitrate is equivalent to 0.005845 g of NaCl. Amoxicillin Oral Suspension
Bacterial endotoxins (2.2.3). Not more than 0.25 Endotoxin Amoxycillin Oral Suspension is a mixture consisting of
Unit per mg of amoxycillin. Amoxycillin Trihydrate with buffering agents and other
Water (2.3.43). Not more than 4.0 per cent, determined on 0.4 excipients. It contains a suitable flavouring agent. It is filled in
g. a sealed container.
Assay. Determine by liquid chromatography (2.4.14). The suspension is constituted by dispersing the contents of
the sealed container in the specified volume of Water just
Solvent mixture. Dissolve 6.8 g of monobasic potassium before issue.
phosphate in 100 ml of water and adjust the pH to about 5.0
with a 4.5 per cent w/v solution of potassium hydroxide. Amoxycillin Oral Suspension contains not less than 90.0 per
Test solution. Determine the weight of the contents of 10 amoxicillin C16H19N3O5S.
containers. Transfer an accurately weighed quantity of the
mixed contents of the 10 containers containing about 100 mg When stored at the temperature and for the period stated on
of amoxycillin to a 100-ml volumetric flask, add 80 ml of the the label during which the constituted suspension may be
solvent mixture and dissolve by shaking and mixing if expected to be satisfactory for use, it contains not less than
necessary, with the aid of ultrasound. Dilute to 100.0 ml with 80.0 per cent of the stated amount of amoxycillin C16H19N3O5S.
the solvent mixture and filter. Use this solution within 6 hours. Storage. Store protected from moisture at a temperature not
Reference solution. Dissolve an accurately weighed quantity exceeding 30°.
of amoxycillin trihydrate RS in the solvent mixture by shaking
Identification
and mixing if necessary, with the aid of ultrasound and dilute
to obtain a solution having a known concentration of about In the Assay, the principal peak in the chromatogram obtained
1.2 mg per ml. Use this solution within 6 hours. with the test solution corresponds to the peak in the
– a stainless steel column 25 cm x 4.0 mm, packed with The constituted suspension complies with the tests stated
octadecylsilane bonded to porous silica or ceramic under Oral liquids and with the following tests.
microparticles (5 µm),
– mobile phase: a mixture of 96 volumes of acetonitrile Tests
and 4 volumes of the solvent mixture,
– flow rate. 1.5 ml per minute, pH (2.4.24). 4.0 to 7.0.
– spectrophotometer set at 230 nm, Assay. Determine by liquid chromatography (2.4.14).
Solvent mixture. Dissolve 6.8 g of monobasic potassium
Inject the reference solution. The test is not valid unless the phosphate in 100 ml of water and adjust the pH to about 4.5
capacity factor is between 1.1 and 2.8, the column efficiency is with a 4.5 per cent w/v solution of potassium hydroxide.
containing 120 mg of amoxicillin to a 100-ml volumetric flask,
dissolve in the solvent mixture and dilute to 100.0 ml with the
Inject alternately the test solution and the reference solution. solvent mixture and filter.
103
AMOXYCILLIN TRIHYDRATE IP 2007
Reference solution. Dissolve an accurately weighed quantity Identification

of amoxycillin trihydrate RS in the solvent mixture by shaking
and mixing if necessary, with the aid of ultrasound and dilute Test A may be omitted if tests B and C are carried out. Tests B
to obtain a solution having a known concentration of about and C may be omitted if test A is carried out.
1.2 mg per ml. Use this solution within 6 hours. A. Determine by infrared absorption spectrophotometry (2.4.6).
Chromatographic system Compare the spectrum with that obtained with amoxycillin
– a stainless steel column 25 cm x 4.0 mm, packed with trihydrate RS or with the reference spectrum of amoxycillin
octadecylsilane chemically bonded to porous silica or trihydrate.
ceramic microparticles (5 µm), B. In the Assay, the principal peak in the chromatogram
– mobile phase: a mixture of 96 volumes of acetonitrile obtained with the test solution corresponds to the peak in the
and 4 volumes of the solvent mixture, chromatogram obtained with the reference solution.
– spectrophotometer set at 230 nm, C. Place about 2 mg in a test-tube. Moisten with 0.05 ml of
– a 10 µl loop injector. water and add 2 ml of sulphuric acid-formaldehyde solution.
Mix the contents of the tube by swirling; the solution is
Inject the reference solution. The test is not valid unless the practically colourless. Place the tube in a water-bath for
capacity factor is between 1.1 and 2.8, the column efficiency is 1 hour; a dark yellow colour develops.
more than 2.5 and the relative standard deviation for replicate Tests
Appearance of solution. Dissolve 1.0 g in 10 ml of 0.5 M
hydrochloric acid, and a further 1.0 g in a mixture of 3 ml of
Determine the weight per ml of the oral suspension (2.4.29) dilute ammonia solution and 7 ml of water. Both solutions
and calculate the content of C16H19N3O5S weight in volume. when freshly prepared are not more opalescent than
Repeat the procedure using a portion of the constituted opalescence standard OS2 (2.4.1).
suspension that has been stored at the temperature and for pH (2.4.24). 3.5 to 5.5, determined in a 0.2 per cent w/v solution.
the period stated on the label.
Specific optical rotation (2.4.22). +290° to +315°, determined
Labelling. The label states (1) the quantity of active ingredient in a 0.2 per cent w/v solution in carbon dioxide-free water.
in terms of the equivalent amount of amoxicillin; (2) the
temperature of storage and the period during which the
determined by Method A.
constituted suspension may be expected to be satisfactory
for use. Heavy metals (2.3.13). 1.0 g complies with the limit test for
Amoxycillin Trihydrate Water (2.3.43). 11.5 to 14.5 per cent, determined on 0.1 g.
H COOH
O Solvent mixture. Dissolve 6.8 g of monobasic potassium
NH2 H N CH3 phosphate in 100 ml of water and adjust the pH to about
N , 3H2O 4.5 with a 4.5 per cent w/v solution of potassium hydroxide.
S CH3
H H Test solution. Transfer an accurately weighed quantity of about
O
HO 120 mg of the substance under examination to a 100-ml
volumetric flask, dissolve in the solvent mixture by shaking
C16H19N3O5S,3H2O Mol. Wt 419.5 and mixing if necessary, with the aid of ultrasound and dilute
to 100.0 ml with the solvent mixture. Use this solution within
Amoxycillin Trihydrate is (6R)-6-(α-4-hydroxyphenyl-D- 6 hours.
glycylamino)penicillanic acid trihydrate.
Amoxycillin Trihydrate contains not less than 95.0 per cent
amoxycillin trihydrate RS, dissolve in the solvent mixture by
and not more than 100.5 per cent of C16H19N3O5S, calculated
and dilute to obtain a solution having a known concentration
Description. A white or almost white, crystalline powder. of about 1.2 mg per ml. Use this solution within 6 hours.
104
IP 2007 AMOXYCILLIN AND POTASSIUM CLAVULANATE INJECTION
Chromatographic system Solvent mixture. Dissolve 6.8 g of monobasic potassium

– a stainless steel column 25 cm x 4.0 mm, packed with phosphate in 100 ml of water and adjust the pH to about
octadecylsilane chemically bonded to porous silica or 4.5 with a 4.5 per cent w/v solution of potassium hydroxide.
ceramic microparticles (5 µm), Test solution. Weigh accurately a quantity of the powdered
– mobile phase: a mixture of 96 volumes of acetonitrile tablets containing about 100 mg of amoxicillin and dissolve in
and 4 volumes of the solvent mixture, the solvent mixture by shaking for 15 minutes and mixing if
– flow rate. 1.5 ml per minute, necessary, with the aid of ultrasound. Dilute to 100.0 ml with
– spectrophotometer set at 230 nm, the solvent mixture and filter. Use this solution within 6 hours.
amoxycillin trihydrate RS, dissolve in the solvent mixture by
and dilute to obtain a solution having a known concentration
of about 1.2 mg per ml. Use this solution within 6 hours.
Calculate the percentage content of C16H19N3O5S. octadecylsilane chemically bonded to porous silica or
Storage. Store at a temperature not exceeding 30°. ceramic microparticles (5 µm),
Labelling. The label states (1) the strength in terms of the and 4 volumes of the solvent mixture,
equivalent amount of amoxycillin; (2) that the tablets should – flow rate. 1.5 ml per minute,
be dispersed in water immediately before use. – spectrophotometer set at 230 nm,
Amoxycillin Dispersible Tablets not less than 1700 theoretical plates, the tailing factor is not
Amoxycillin Trihydrate Dispersible Tablets; Dispersible more than 2.5 and the relative standard deviation for replicate
Amoxicillin Tablets injections is not more than 2.0 per cent.
Amoxycillin Dispersible Tablets contain Amoxycillin Inject alternately the test solution and the reference solution.
Trihydrate in a suitable dispersible base. Calculate the content of C16H19N3O5S in the tablets.
Amoxycillin Dispersible Tablets contain not less than 90.0 per Storage. Store protected from moisture at a temperature not
cent and not more than 120.0 per cent of the stated amount of exceeding 30°.
amoxycillin, C16H19N3O5S.
Identification
Shake a quantity of the powdered tablets containing 0.5 g of Amoxycillin and Potassium
amoxycillin with 5 ml of water for 5 minutes, filter, wash the
residue first with ethanol and then with ether and dry for Clavulanate Injection
1 hour at a pressure not exceeding 0.7 kPa. The residue Amoxicillin and Potassium Clavulanate injection
complies with the following test.
Amoxycillin and Potassium Clavulanate injection is a sterile
Determine by infrared absorption spectrophotometry (2.4.6). material consisting of Amoxycillin Sodium and Potassium
Compare the spectrum with that obtained with amoxycillin Clavulanate with or without excipients. It is filled in a sealed
trihydrate RS or with the reference spectrum of amoxycillin container.
trihydrate.
The injection is constituted by dissolving the contents of the
Tests sealed container in the requisite amount of sterile Water for
Assay. Weigh and powder 20 tablets. Clarity of solution and Particulate matter stated under
Determine by liquid chromatography (2.4.14). Parenteral Preparations (Injections).
105
AMOXYCILLIN AND POTASSIUM CLAVULANATE ORAL SUSPENSION IP 2007
Storage. The constituted solution should be used immediately Assay. Determine by liquid chromatography (2.4.14).
after preparation but, in any case, within the period Test solution. Determine the weight of the contents of 10
recommended by the manufacturer. containers. Dissolve, with shaking, a quantity of the mixed
Amoxycillin and Potassium Clavulanate Injection contains not contents of the 10 containers containing about 60 mg of
less than 90.0 per cent and not more than 107.5 per cent of the amoxycillin in water and dilute to 100.0 ml with the same
stated amounts of amoxycillin,C16H19N3O5S and of clavulanic solvent, mix and filter.
acid, C8H9NO5.
Reference solution. A solution containing 0.06 per cent w/v of
The contents of the sealed container comply with the amoxycillin trihydrate RS and 0.012 per cent w/v of clavulanic
requirements stated under Parenteral Preparations acid RS in water.
(Powders for Injection) and with the following requirements. Chromatographic system
– a stainless steel column 30 cm × 3.9 mm, packed with
Identification octadecylsilane bonded to porous silica (10 µm),
A. Determine by thin-layer chromatography (2.4.17), coating – mobile phase: a mixture of 95 volumes of buffer solution
the plate with silica gel F254 (such as Merck silica gel 60 prepared by dissolving 7.8 g of monobasic sodium
F254 plates). phosphate in 900 ml of water, adjust the pH to 4.4 with
10 M sodium hydroxide or orthophosphoric acid and
Mobile phase. A mixture of 1 volume of butan-1-ol , 2 volumes 5 volumes of methanol,
of a 0.1 per cent w/v solution of disodium edetate in mixed – flow rate. 2 ml per minute,
phosphate buffer pH 4.0, 6 volumes of glacial acetic acid – spectrophotometer set at 220 nm,
and 10 volumes of butyl acetate. – a 20 µl loop injector.
Test solution. Shake a quantity of the contents of the sealed Inject the reference solution. The test is not valid unless the
container containing 0.4 g of clavulanic acid in 100 ml of a resolution between the peaks due to amoxycillin and clavulanic
mixture of 4 volumes of methanol and 6 volumes of 0.1 M acid is not less than 3.5, the tailing factor is not more than 1.5,
mixed phosphate buffer pH 7.0 and filter. the column efficiency is not less than 550 theoretical plates
Reference solution. A solution containing 0.4 per cent w/v of for both component and the relative standard deviation is not
lithium clavulanate RS and 0.8 per cent w/v of amoxycillin more than 2.0.
trihydrate RS in a mixture of 4 volumes of methanol and 6 Inject alternately the test solution and the reference solution.
volumes of 0.1 M mixed phosphate buffer pH 7.0. Calculate the content of C16H19N3O5S and C8H9NO5.
Apply to the plate 1 µl of each of the solutions after 1 mg of C8H8LiNO5 is equivalent to 0.9711 mg of C8H9NO5.
impregnating the plate by spraying it with a 0.1 per cent w/v
Labelling. The label states the quantity of Amoxycillin Sodium
solution of disodium edetate in mixed phosphate buffer pH
contained in it, in terms of the equivalent amount of
4.0 and allowing to dry overnight and activating the plate by
amoxycillin, and the quantity of Potassium Clavulanate, in
heating at 105º for 1 hour just before use. After development,
terms of the equivalent amount of clavulanic acid.
allow it to dry in air and examine in ultraviolet light at 254 nm.
The principal spots in the chromatogram obtained with the
test solution correspond to those in the chromatogram
obtained with the reference solution. Amoxycillin and Potassium
B. In the Assay, the retention time of the two principal peaks Clavulanate Oral Suspension
in the chromatogram obtained with the test solution Amoxicillin and Potassium Clavulanate oral suspension
correspond to those in the chromatogram obtained with the
reference solution. Amoxycillin and Potassium Clavulanate oral suspension is a
mixture of Amoxycillin Tryhydrate and Potassium Clavulanate
Tests with buffering agents and other excipients. It contains a
suitable flavouring agent.
pH (2.4.24). 8.0 to 10.0, determined in a solution containing 10
The suspension is constituted by dispersing the contents of
per cent w/v of amoxycillin.
Bacterial endotoxins (2.2.3). Not more than 0.25 EU per mg of before use.
amoxycillin.
Amoxycillin and Potassium Clavulanate Oral Suspension
Water (2.3.43). Not more than 3.5 per cent, determined on contains not less than 90.0 per cent and not more than 120.0
0.5 g. per cent of the stated amount of amoxycillin, C16H19N3O5S and
106
IP 2007 AMOXYCILLIN AND POTASSIUM CLAVULANATE TABLETS
not less than 90.0 per cent and not more than 125.0 per cent of resolution between the amoxycillin and clavulanic acid peaks
the stated amount of clavulanic acid, C8H9NO5. is not less than 3.5. The test is not valid unless the column
When stored at the temperature and for the period stated on efficiency determined from each analyte peak is not less than
the label during which the constituted suspension may be 550 theoretical plates, the tailing factor for each analyte peak
expected to be satisfactory for use, it contains not less than is not more than 1.5 and the relative standard deviation for
80.0 per cent of the stated amounts of amoxycillin, C16H19N3O5S replicate injections is not more than 2.0 per cent.
and clavulanic acid, C8H9NO5. Inject alternately the test solution and the reference solution.
Determine the weight per ml of the oral suspension (2.4.29)
Identification
and calculate the content of C16H19N3O5S and C8H9NO5 weight
In the Assay, the retention time of the two principal peaks in in volume.
the chromatogram obtained with the test solution correspond 1 mg of C8H8LiNO5 is equivalent to 0.9711 mg of C8H9NO5.
to those in the chromatogram obtained with the reference
solution. Repeat the procedure using a portion of the constituted
suspension that has been stored at the temperature and for
Tests the period stated on the label.
Water (2.3.43). Not more than 7.5 per cent where the label Storage. Store protected from moisture.
indicates that after reconstitution as directed, the suspension Labelling. The label states the quantity of Amoxycillin
contains an amount of amoxycillin that is less than 40 mg per Trihydrate contained in it, in terms of the equivalent amount
ml; not more than 8.5 per cent where the label indicates that of amoxycillin, and the quantity of Potassium Clavulanate, in
after reconstitution as directed, the suspension contains an terms of the equivalent amount of clavulanic acid.
amount of amoxicillin that is equal to or more than 40 mg per ml
and is less than or equal to 50 mg per ml; not more than 11.0
per cent where the label indicates that after reconstitution as
directed, the suspension contains an amount of amoxycillin
Amoxycillin and Potassium
that is more than 50 mg per ml and is less than or equal to 80 Clavulanate Tablets
mg per ml; not more than 12.0 per cent where the label indicates
that after reconstitution as directed, the suspension contains Amoxicillin and Potassium Clavulanate Tablets
an amount of amoxycillin that is more than 80 mg per ml. Amoxycillin and Potassium Clavulanate Tablets contain
The constituted suspension complies with the tests stated Amoxycillin Trihydrate and Potassium Clavulanate.
under Oral liquids and with the following tests. Amoxycillin and Potassium Clavulanate Tablets contain not
pH (2.4.24). 3.8 to 6.6. less than 90.0 per cent and not more than 120.0 per cent of the
stated amounts of amoxycillin, C16H19N3O5S and clavulanic
Assay. Determine by liquid chromatography (2.4.14). acid, C8H9NO5.
Identification
containing about 50 mg of amoxycillin to a 100-ml volumetric
flask, dissolve in water, dilute to 100.0 ml with the same solvent In the Assay, the retention time of the two principal peaks in
and filter. Use the filtrate as the test solution within 1 hour. the chromatogram obtained with the test solution correspond
Reference solution. A solution containing 0.05 per cent w/v of to those in the chromatogram obtained with the reference
amoxycillin trihydrate RS and 0.02 per cent w/v of lithium solution.
clavulanate RS in water.
Tests
Disintegration (2.5.1). 30 minutes, for tablets labelled for
– a stainless steel column 30 cm x 4 mm, packed with
veterinary use only, simulated gastric fluid being substituted
octadecylsilane bonded to porous silica (3 to 10 µm),
for water in the test.
– mobile phase: a mixture of 95 volumes of pH 4.4 sodium
phosphate buffer and 5 volumes of methanol, Dissolution (2.5.2). (Tablets labelled for veterinary use only
– flow rate. 2 ml per minute, are exempt from this requirement).
– spectrophotometer set at 220 nm, Apparatus. No 1
– a 20 µl loop injector. Medium. 900 ml of water.
Inject the reference solution. The relative retention times are Speed and time. 75 rpm and 30 minutes or 45 minutes where
about 0.5 for clavulanic acid and 1.0 for amoxycillin. The the Tablets are labelled as chewable.
107
AMPHOTERICIN B IP 2007
Withdraw a suitable volume of the medium and filter. Carry Inject the reference solution. The relative retention times are
out the method described under Assay. about 0.5 for clavulanic acid and 1.0 for amoxycillin. The
D. Not less than 85 per cent of the stated amount of resolution between the amoxycillin and clavulanic acid peaks
C16H19N3O5S and not less than 80 per cent of the stated is not less than 3.5. The test is not valid unless the column
amount of C8H9NO5. efficiency determined from each analyte peak is not less than
550 theoretical plates, the tailing factor for each analyte peak
For tablets labelled as chewable. Not less than 80 per cent of is not more than 1.5 and the relative standard deviation for
the stated amount of the C 16H19N 3O 5S and C 8H9NO 5 is replicate injections is not more than 2.0 per cent.
dissolved in 45 minutes.
Calculate the content of C16H19N3O5S and C8H9NO5 in the
Tablets, determining the content of clavulanic acid in the
tablets.
tablets.
Follow the chromatographic procedure described under Assay 1 mg of C8H8LiNO5 is equivalent to 0.9711 mg of C8H9NO5.
using the following test solution. Storage. Store protected from moisture.
Powder one tablet and transfer to a 100 ml flask. Dissolve in Labelling. The label includes the word “chewable” in
water and dilute to 100.0 ml with the same solvent and filter. juxtaposition to the official name in the case of Chewable
Further dilute to obtain a solution containing 0.05 per cent w/ Tablets. The label also indicates that Chewable Tablets may
v of amoxycillin. Use the solution within 1 hour. be chewed before being swallowed or may be swallowed
whole. Tablets intended for veterinary use only are so labelled.
Calculate the content of C8H9NO5 in the tablet.
Water (2.3.43). Not more than 7.5 per cent, where the labelled
amount of amoxycillin in each tablet is 250 mg or less; not
more than 10.0 per cent where the labelled amount of Amphotericin B
amoxycillin in each tablet is more than 250 mg but less than or
equal to 500 mg; not more than 11.0 per cent where the labelled
OH
amount of amoxycillin in each tablet is more than 500 mg. OH
H3C O OH
Where the tablets are labelled as chewable, not more than 6.0
per cent where the labelled amount of amoxycillin in each HO O OH OH OH OH O
CH3 COOH
tablet is 125 mg or less; not more than 8.0 per cent where the H
labelled amount of amoxycillin in each tablet is more than 125 H3C
mg. Where the tablets are labelled for veterinary use only, not O O CH3
OH
more than 10.0 per cent. NH2
OH
Assay. Determine by liquid chromatography (2.4.14). C47H73NO17 Mol. Wt. 924.1
Test solution. Weigh and powder 20 tablets. Weigh accurately Amphotericin B is a mixture consisting mainly of
a quantity of the powdered tablet containing about 50 mg of amphotericin B which is
amoxycillin, dissolve in water, dilute to 100.0 ml with water (3R,5R,8R,9R,11S,13R,15S,16R,17S,19R,34S,35R,36R,37S)-19-
and filter. Use the filtrate as the test solution within 1 hour. (3-amino-3,6-dideoxy-β-D-mannopyranosyloxy)-16-carboxy-
3,5,8,9,11,13,15, 35-octahydroxy-34,36-dimethyl-13,17-
epoxyoctatriaconta- 20,22,24,26,28,30,32-heptaen-37-olide
of amoxycillin trihydrate RS and 0.02 per cent w/v of lithium
and other antifungal polyenes produced by the growth of
clavulanate RS in water.
certain strains of Streptomyces nodosus or by any other
Chromatographic system means.
Amphotericin B has a potency of not less than 750 Units per
octadecylsilane bonded to porous silica (3 to 10 µm),
mg, calculated on the dried basis.
– mobile phase: a mixture of 95 volumes of 0.78 per cent
w/v solution of sodium phosphate, adjusted to pH 4.4 Description. A yellow to orange powder; practically odourless.
with orthophosphoric acid and 5 volumes of methanol, Even in the absence of light, it is gradually decomposed in a
– flow rate. 2 ml per minute, humid environment, degradation being faster at higher
– spectrophotometer set at 220 nm, temperatures. In solutions, it is inactivated in the presence of
– a 20 µl loop injector. light and at low pH values.
108
IP 2007 AMPHOTERICIN B INJECTION
Identification Sulphated ash (2.3.18). Not more than 3.0 per cent; for
parenteral use, not more than 0.5 per cent.
Compare the spectrum with that obtained with amphotericin Loss on drying (2.4.19). Not more than 5.0 per cent, determined
B RS or with the reference spectrum of amphotericin B. on 1.0 g by drying in an oven at 60° at a pressure not exceeding
0.7 kPa.
B. Dissolve 25 mg in 5 ml of dimethyl sulphoxide, add sufficient
methanol to produce 50 ml, and dilute 2 ml to 200 ml with Assay. Determine by the microbiological assay of antibiotics,
methanol. When examined in the range 300 nm to 450 nm Method A (2.2.10) on a solution prepared in the following
(2.4.7), the resulting solution shows absorption maxima at about manner. Weigh accurately about 60 mg, triturate with
362 nm, 381 nm, and 405 nm. The ratio of the absorbance at the dimethylformamide and add, with shaking, sufficient
maximum at about 362 nm to the absorbance at the maximum at dimethylformamide to produce 100.0 ml. Dilute 10.0 ml to
about 381 nm, 0.5 to 0.6; the ratio of the absorbance at the 100.0 ml with dimethylformamide. Express the result in Units
maximum at about 381 nm to the absorbance at the maximum at per mg.
about 405 nm, about 0.9. Amphotericin B intended for use in the manufacture of
C. To 1 ml of a 0.05 per cent w/v solution in dimethyl sulphoxide parenteral preparations without a further appropriate
add 5 ml of phosphoric acid to form a lower layer; a blue ring procedure for removal of bacterial endotoxins complies with
is immediately formed at the junction of the liquids. Mix; the the following additional requirement.
mixture becomes intensely blue. Add 15 ml of water and mix; Bacterial endotoxins (2.2.3). Not more than 1.0 Endotoxin Unit
the solution becomes pale straw-coloured. per mg, using the supernatant liquid obtained after shaking
50 mg with 25 ml of water BET and centrifuging.
Tests
Amphotericin B intended for use in the manufacture of
pH (2.4.24). 6.0 to 8.0, determined in a 3.0 per cent w/v parenteral preparations without a further appropriate
suspension in water; for parenteral use, 3.5 to 6.0. sterilisation procedure complies with the following
Tetraenes. Not more than 15.0 per cent (for parenteral use, not additional requirement.
more than 10.0 per cent), determined by the following method. Sterility (2.2.11). Complies with the test for sterility, using
Weigh accurately about 50 mg, dissolve in 5 ml of dimethyl 50 mg from each container.
sulphoxide, dilute to 50.0 ml with methanol and dilute 4.0 ml
of the resulting solution to 50.0 ml with methanol (solution 1). Storage. Store protected from light in a refrigerator (2° to 8°).
Prepare solution (2) in a similar manner using 50 mg of Do not freeze.
amphotericin B RS, accurately weighed, instead of the Labelling. The label states (1) the number of Units per mg; (2)
substance under examination. For solution (3) dissolve 25 mg whether the material is intended for use in the manufacture of
of nystatin RS, accurately weighed, in 25 ml of dimethyl parenteral preparations.
sulphoxide, dilute to 250.0 ml with methanol and dilute 4.0 ml
to 50.0 ml with methanol. Using as the blank a 0.8 per cent
v/v solution of dimethyl sulphoxide in methanol, measure Amphotericin B Injection
the absorbances of solutions (1), (2) and (3) at the maxima at
Amphotericin B Injection is a sterile freeze dried mixture of
about 282 nm and about 304 nm (2.4.7).
Amphotericin B and deoxycholate sodium with one or more
Calculate the specific absorbances for the substance under buffering agents. It is filled in a sealed container.
examination, amphotericin B RS and nystatin RS at both
wavelengths and calculate the content of tetraenes from the
sealed container in the requisite amount of sterile Water for
expression
25 WN [(A B282 × A U304 ) − (A B304 × A U282 ) The constituted solution complies with the requirements for
[(A B282 × A N304 ) − (A B304 × A N282 )] WU Clarity of solution and Particulate Matter stated under
Parenteral Preparations (Injections)
where WN is the weight, in mg, of nystatin RS, AB282 and AB304
are the specific absorbances of amphotericin B RS at about Storage. The constituted solution should be used immediately
282 nm and 304 nm, respectively, AN282 and AN304 are the specific after preparation but, in any case within the period
absorbances of nystatin RS at about 282 nm and 304 nm recommended by the manufacturer.
respectively, AU282 and AU304 are the specific absorbances of Amphotericin B Injection contains not less than 90.0 per cent
the substance under examination at about 282 nm and 304 nm and not more than 120.0 per cent of the stated amount of
respectively and WU is the weight in mg of the sample taken. amphotericin B, C47H73NO17.
109
AMPICILLIN IP 2007
The contents of the sealed container comply with the B. In the Assay, the principal peak in the chromatogram
requirements stated under Parenteral Preparations obtained with the test solution corresponds to the peak in the
(Powders for Injection) and with the following requirements. chromatogram obtained with reference solution (a).
Tests Tests
pH (2.4.24). 7.2 to 8.0 determined in a solution containing 50
Appearance of solution. Dissolve 1.0 g in 10 ml of 1 M
mg per ml of Amphotericin B.
hydrochloric acid and a further 1.0 g in a mixture of 3 ml of
Bacterial Endotoxins (2.2.3). Not more than 5.0 Endotoxin unit dilute ammonia solution and 7 ml of water. Both solutions
per mg of amphotericin B. For products used or labelled for when freshly prepared are not more opalescent than
intrathecal injection, not more than 0.9 Endotoxin unit per mg. opalescence standard OS2 (2.4.1).
Loss on drying (2.4.19). Not more than 8.0 per cent, determined pH (2.4.24). 3.5 to 5.5, determined in a 0.25 per cent w/v solution.
on 0.1 g by drying in an oven at 60° at a pressure not exceeding
0.7 kPa.
in a 0.25 per cent w/v solution.
N, N-Dimethylaniline (2.3.21). Not more than 20 ppm.
Method A (2.2.10) on a solution prepared in the following
determined by Method B.
manner.
Mix the contents of 10 containers, dissolve in
dimethylformamide. Express the results in mg per vial, taking
each 1000 units found to be equivalent to 1 mg of amphotericin Sulphated ash (2.3.18). Not more than 0.5 per cent.
B. Water (2.3.43). Not more than 2.0 per cent, determined on
Storage. Store in tightly closed containers between 2° to 8°, 0.3 g.
protected from light.
Assay. Determine by liquid chromatography (2. 4.14).
Labeling. Label it to state that it is intended for use by
Solvent mixture. Mix 10 ml of 1 M monobasic potassium
intravenous infusion to hospitalised patients only, and that
phosphate and 1 ml of 1 M acetic acid and dilute to 1000 ml
the solution should be protected from light during
with water.
administration.
containing about 100 mg of ampicillin to a 100-ml volumetric
Ampicillin flask, add about 80 ml of the solvent mixture, shake and mix
with the aid of ultrasound if necessary to achieve complete
H COOH dissolution and dilute to 100.0 ml with the solvent mixture.
O Use this solution promptly after preparation.
NH2 N CH3
H
N Reference solution (a). Weigh accurately a suitable quantity
S CH3
of ampicillin RS, dissolve in the solvent mixture by shaking
H H
O and mixing if necessary, with the aid of ultrasound to obtain a
solution having a known concentration of about 1 mg per ml.
C16H19N3O4S Mol. Wt. 349.4 Use this solution promptly after preparation.
Ampicillin is (6R)-6-(α-phenyl-D-glycylamino)penicillanic Reference solution (b). Dissolve caffeine in reference solution
acid. (a) to obtain a solution containing about 0.12 mg per ml.
Ampicillin contains not less than 96.0 per cent and not more Chromatographic system
than 100.5 per cent of C16H 19N 3O 4S, calculated on the – a stainless steel column 30 cm x 4.0 mm, packed with
anhydrous basis. octadecylsilane chemically bonded to porous silica or
Description. A white, crystalline powder.
– mobile phase: a mixture of 900 volumes of water, 80
Identification volumes of acetonitrile, 10 volumes of 1 M monobasic
potassium phosphate, and 1 ml of 1 M acetic acid,
A. Determine by infrared absorption spectrophotometry (2.4.6). – flow rate. 2 ml per minute,
Compare the spectrum with that obtained with ampicillin RS – spectrophotometer set at 254 nm,
or with the reference spectrum of ampicillin. – a 20 µl loop injector.
110
IP 2007 AMPICILLIN SODIUM
Inject reference solution (b). The resolution between the Test solution. Weigh accurately a quantity of the mixed
caffeine and ampicillin peaks is not less than 2.0. The relative contents of 20 capsules containing about 100 mg of ampicillin,
retention times are about 0.5 for ampicillin and 1.0 for caffeine. add about 80 ml of the solvent mixture and dissolve by shaking
Inject reference solution (a). The test is not valid unless the for 15 minutes and mixing if necessary, with the aid of
capacity factor is not more than 2.5, the tailing factor is not ultrasound. Dilute to 100.0 ml with the solvent mixture and
more than 1.4 and the relative standard deviation for replicate filter. Use this solution promptly after preparation.
injections is not more than 2.0 per cent. Reference solution (a). Weigh accurately a suitable quantity
Inject alternately the test solution and reference solution (a).
and mixing if necessary, with the aid of ultrasound to obtain a
Calculate the percentage content of C16H19N3O4S. solution having a known concentration of about 1 mg per ml.
Storage. Store protected from moisture at a temperature not Use this solution promptly after preparation.
exceeding 30°. Reference solution (b). Dissolve caffeine in reference solution
(a) to obtain a solution containing about 0.12 mg per ml.
Ampicillin Capsules – a stainless steel column 30 cm x 4.0 mm, packed with
Ampicillin Capsules contain Ampicillin or Ampicillin Trihydrate octadecylsilane chemically bonded to porous silica or
equivalent to not less than 92.5 per cent and not more than ceramic micro particles (5 µm),
107.5 per cent of the stated amount of ampicillin, C16H19N3O4S. – mobile phase: a mixture of 900 volumes of water,
80 volumes of acetonitrile, 10 volumes of 1 M
Identification monobasic potassium phosphate, and 1 ml of 1 M acetic
acid,
The contents of the capsules comply with the following tests.
A. Determine by infrared absorption spectrophotometry (2.4.6). – spectrophotometer set at 254 nm,
Compare the spectrum with that obtained with ampicillin RS – a 20 µl loop injector.
or with the reference spectrum of ampicillin. Inject reference solution (b). The resolution between the
B. In the Assay, the principal peak in the chromatogram caffeine and ampicillin peaks is not less than 2.0. The relative
obtained with the test solution corresponds to the peak in the retention times are about 0.5 for ampicillin and 1.0 for caffeine.
chromatogram obtained with reference solution (a). Inject reference solution (a). The capacity factor is not more
than 2.5 and the tailing factor is not more than 1.4. The test is
Tests
not valid unless the relative standard deviation for replicate
Dissolution (2.5.2). injections is at most 2.0 per cent.
Apparatus. No 1 Inject alternately the test solution and reference solution (a).
Medium. 900 ml of water. Calculate the content of C16H19N3O4S in the capsules.
Speed and time. 100 rpm and 45 minutes. Storage. Store protected from moisture at a temperature not
Use one capsule in the vessel for each test. exceeding 30°.
Withdraw a suitable volume of the medium and filter promptly. Labelling. The label states the strength in terms of the
Dilute the filtrate, if necessary, with the same solvent. Measure equivalent amount of ampicillin (when Ampicillin Trihydrate
the absorbance (2.4.7) of the resulting solution at the maximum is used).
at about 272 nm. Calculate the content of C16H19N3O4S in the
medium from the absorbance obtained from a solution of
known concentration of ampicillin RS. Ampicillin Sodium
D: Not less than 80 per cent of the stated amount of
C16H19N3O4S. H COONa
O
Other tests. Comply with the tests stated under Capsules. NH2 N CH3
H
Assay. Determine by liquid chromatography (2. 4.14). N S CH3
H H
Solvent mixture. Mix 10 ml of 1 M monobasic potassium O
with water. C16H18N3NaO4S Mol. Wt. 371.4
111
AMPICILLIN SODIUM IP 2007
Ampicillin Sodium is sodium (6R)-6-(α-phenyl-D-glycyl- – inlet port and detector.150°,

amino)penicillinate – flow rate. 40 ml per minute of the carrier gas.
Ampicillin Sodium contains not less than 92.5 per cent and Calculate the percentage w/w of dichloromethane, assuming
not more than 100.5 per cent of C16H18N3NaO4S, calculated on its relative density (2.4.29) to be 1.325 g.
the anhydrous basis. Heavy metals (2.3.13). 1.0 g complies with the limit test for
Description. A white, crystalline powder; hygroscopic. heavy metals, Method B (20 ppm).
Identification
Assay. Determine by liquid chromatography (2. 4.14).
A. Determine by infrared absorption spectophotometry (2.4.6).
Compare the spectrum with that obtained with ampicillin Solvent mixture. Mix 10 ml of 1 M monobasic potassium
sodium RS or with the reference spectrum of ampicillin sodium. phosphate and 1 ml of 1 M acetic acid and dilute to 1000 ml
with water.
B. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak in the Test solution. Transfer an accurately weighed quantity
chromatogram obtained with reference solution (a). containing about 100 mg of ampicillin to a 100-ml volumetric
flask and dissolve in the solvent mixture by shaking and mixing
C. A 5 per cent w/v solution gives the reactions of sodium if necessary, with the aid of ultrasound and dilute to 100.0 ml
salts (2.3.1). with the solvent mixture. Use this solution promptly after
preparation.
Tests
Reference solution (a). Weigh accurately a suitable quantity
Appearance of solution. A 10.0 per cent w/v solution is clear, of ampicillin RS, dissolve in the solvent mixture by shaking
when examined immediately after preparation (2.4.1), and the and mixing if necessary, with the aid of ultrasound to obtain a
absorbance of the solution at about 430 nm is not more than solution having a known concentration of about 1 mg per ml.
0.15 (2.4.7). Use this solution promptly after preparation.
pH (2.4.24). 8.0 to 10.0, determined 10 minutes after dissolution Reference solution (b). Dissolve caffeine in reference solution
in a 10.0 per cent w/v solution. (a) to obtain a solution containing about 0.12 mg per ml.
Specific optical rotation (2.4.22). +258° to +287°, determined Chromatographic system
in a 0.25 per cent w/v solution in a 0.4 per cent w/v solution of – a stainless steel column 30 cm x 4.0 mm, packed with
potassium hydrogen phthalate. octadecylsilane chemically bonded to porous silica or
N,N-Dimethylaniline (2.3.21). Not more than 20 ppm, ceramic microparticles (5 µm),
determined by Method B. – mobile phase: a mixture of 900 volumes of water,
Dichloromethane. Not more than 0.2 per cent w/w, determined
monobasic potassium phosphate, and 1 ml of 1 M acetic
in the following manner.
acid,
Determine by gas chromatography (2.4.13). – flow rate. 2 ml per minute,
Internal standard solution. A 0.02 per cent v/v solution of – spectrophotometer set at 254 nm,
1,2-dichloroethane in water. – a 20 µl loop injector.
Test solution. Dissolve 1.0 g of the substance under Inject reference solution (b). The resolution between the
examination in sufficient water to produce 10 ml. caffeine and ampicillin peaks is not less than 2.0. The relative
retention times are about 0.5 for ampicillin and 1.0 for caffeine.
Reference solution (a). Dissolve 1.0 g of the substance under
examination in 10 ml of the internal standard solution. Inject reference solution (a). The test is not valid unless the
capacity factor is not more than 2.5, the tailing factor is not
Reference solution (b). Mix equal volumes of the internal more than 1.4 and the relative standard deviation for replicate
standard solution and a 0.02 per cent v/v solution of injections is not more than 2.0 per cent.
dichloromethane in water.
– a glass column 1.5 m x 5 mm, packed with acid-washed Calculate the percentage content of C16H19N3O4S.
silanised diatomaceous support (100 to 120 mesh) coated Ampicillin Sodium intended for use in the manufacture of
with 10 per cent w/w of polyethylene glycol 1000, parenteral preparations without a further appropriate
– temperature: procedure for the removal of bacterial endotoxins complies
column. 60°, with the following additional requirement.
112
IP 2007 AMPICILLIN INJECTION
Bacterial endotoxins (2.2.3). Not more than 0.15 Endotoxin Tests

Unit per mg.
Appearance of solution. A 10 per cent w/v solution is clear,
Ampicillin Sodium intended for use in the manufacture of when examined immediately after preparation (2.4.1), and the
parenteral preparations without a further appropriate absorbance of the solution at about 430 nm is not more than
sterilization procedure complies with the following 0.15.
additional requirement.
pH (2.4.24). 8.0 to 10.0, determined 10 minutes after dissolution
Sterility (2.2.11). Complies with the test for sterility. in a 10 per cent w/v solution.
Storage. Store protected from moisture at a temperature not Specific optical rotation (2.4.22). +258° to +287°, determined
exceeding 30°. If it is intended for use in the manufacture of in a 0.25 per cent w/v solution in a 0.4 per cent w/v solution of
parenteral preparations, the container should be sterile, tamper- potassium hydrogen phthalate.
evident and sealed so as to exclude micro-organisms.
Labelling. The label states whether or not the material is determined by Method B.
intended for use in the manufacture of parenteral preparations.
Dichloromethane. Not more than 0.2 per cent w/w, determined
in the following manner.
Ampicillin Injection Determine by gas chromatography (2.4.13).

Internal standard solution. A 0.02 per cent v/v solution of
Ampicillin Sodium Injection
1,2-dichloroethane in water.
Ampicillin Injection is a sterile material consisting of Ampicillin
Sodium with or without buffering agents and other excipients.
examination in sufficient water to produce 10 ml.
It is filled in a sealed container.
examination in 10 ml of the internal standard solution.
Injections, immediately before use. Reference solution (b). Mix equal volumes of the internal
standard solution and a 0.02 per cent v/v solution of
dichloromethane in water.
Clarity of solution and Particulate matter stated under
Parenteral Preparations (Injections). Chromatographic system
Storage. The constituted solution should be used immediately – a glass column 1.5 m x 5 mm, packed with acid-washed
after preparation but, in any case, within the period silanised diatomaceous support (100 to 120 mesh) coated
recommended by the manufacturer. with 10 per cent w/w of polyethylene glycol 1000,
– temperature:
Ampicillin Injection contains not less than 95.0 per cent and column. 60°,
not more than 105.0 per cent of the stated amount of ampicillin, – inlet port and detector.150°,
C16H19N3O4S. – flow rate. 40 ml per minute of the carrier gas.
Description. A white or almost white powder; hygroscopic. Calculate the percentage w/w of dichloromethane, assuming
The contents of the sealed container comply with the its relative density (2.4.29) to be 1.325 g.
requirements stated under Parenteral Preparations Heavy metals (2.3.13). 1.0 g complies with the limit test for
(Powders for Injections) and with the following requirements. heavy metals, Method B (20 ppm).
Identification Bacterial endotoxins (2.2.3). Not more than 0.15 Endotoxin
Unit per mg of ampicillin.
A. Determine by infrared absorption spectophotometry (2.4.6).
Compare the spectrum with that obtained with ampicillin Sterility (2.2.11). Complies with the test for sterility.
sodium RS or with the reference spectrum of ampicillin sodium. Water (2.3.43). Not more than 2.0 per cent, determined on
B. In the Assay, the principal peak in the chromatogram 0.3 g.
obtained with the test solution corresponds to the peak in the Assay. Determine by liquid chromatography (2. 4.14).
chromatogram obtained with reference solution (a).
C. A 5 per cent w/v solution gives the reactions of sodium phosphate and 1 ml of 1 M acetic acid and dilute to 1000 ml
salts (2.3.1). with water.
113
AMPICILLIN ORAL SUSPENSION IP 2007
Test solution. Determine the weight of the contents of The suspension is constituted by dispersing the contents of
10 containers. Transfer an accurately weighed quantity of the the sealed container in the specified volume of Water just
mixed contents of the 10 containers containing about 100 mg before issue.
of ampicillin to a 100-ml volumetric flask, add about 80 ml of Ampicillin oral suspension contains not less than 90.0 per
the solvent mixture and dissolve by shaking and mixing if cent and not more than 120.0 per cent of the stated amount of
necessary, with the aid of ultrasound. Dilute to 100.0 ml with ampicillin, C16H19N3O4S.
the solvent mixture and filter. Use this solution promptly after
preparation. The constituted suspension, when stored at the temperature
and for the period stated on the label during which it may be
Reference solution (a). Weigh accurately a suitable quantity expected to be satisfactory for use, contains not less than
of ampicillin RS, dissolve in the solvent mixture by shaking 80.0 per cent of the stated amount of ampicillin, C16H19N3O4S.
solution having a known concentration of about 1 mg per ml. Storage. Store protected from moisture at a temperature not
Use this solution promptly after preparation. exceeding 30°.
Reference solution (b). Dissolve caffeine in reference solution Identification

(a) to obtain a solution containing about 0.12 mg per ml. In the Assay, the principal peak in the chromatogram obtained
Chromatographic system with the test solution corresponds to the peak in the
– a stainless steel column 30 cm x 4.0 mm, packed with chromatogram obtained with reference solution (a).
octadecylsilane chemically bonded to porous silica or The constituted suspension complies with the tests stated
ceramic microparticles (5 µm), under Oral liquids and with the following tests.
– mobile phase: a mixture of 900 volumes of water,
pH (2.4.24). 4.0 to 7.0.
monobasic potassium phosphate, and 1 ml of 1 M acetic Assay. Determine by liquid chromatography (2. 4.14).
acid, Solvent mixture. Mix 10 ml of 1 M monobasic potassium
– flow rate. 2 ml per minute, phosphate and 1 ml of 1 M acetic acid and dilute to 1000 ml
– spectrophotometer set at 254 nm, with water.
Inject reference solution (b). The resolution between the containing about 100 mg of ampicillin to a 100-ml volumetric
caffeine and ampicillin peaks is not less than 2.0. The relative flask and dilute to 100.0 ml with the solvent mixture and filter.
retention times are about 0.5 for ampicillin and 1.0 for caffeine. Use this solution promptly after preparation.
Inject reference solution (a). The test is not valid unless the Reference solution (a). Weigh accurately a suitable quantity
capacity factor is not more than 2.5, the tailing factor is not of ampicillin RS, dissolve in the solvent mixture by shaking
more than 1.4 and the relative standard deviation for replicate and mixing with the aid of ultrasound if necessary, to obtain a
injections is not more than 2.0 per cent. solution having a known concentration of about 1 mg per ml.
Inject alternately the test solution and reference solution (a). Use this solution promptly after preparation.
Reference solution (b). Dissolve caffeine in reference solution
Calculate the content of C16H19N3O4S in the injection.
Storage. Store protected from moisture, in a sterile, tamper-
evident container sealed so as to exclude micro-organisms, at
a temperature not exceeding 30°.
Labelling. The label states the quantity of Ampicillin Sodium ceramic microparticles (5 µm),
contained in the sealed container in terms of the equivalent – mobile phase: a mixture of 900 volumes of water,
amount of anhydrous ampicillin. 80 volumes of acetonitrile, 10 volumes of 1 M
acid,
Ampicillin Oral Suspension – spectrophotometer set at 254 nm,
Ampicillin Oral Suspension is a mixture consisting of Ampicillin – a 20 µl loop injector.
or Ampicillin Trihydrate with buffering agents and other Inject reference solution (b). The resolution between the
excipients. It contains a suitable flavouring agent. It is filled in caffeine and ampicillin peaks is not less than 2.0. The relative
a sealed container. retention times are about 0.5 for ampicillin and 1.0 for caffeine.
114
IP 2007 AMPICILLIN TRIHYDRATE
Inject reference solution (a). The test is not valid unless the solvent mixture and filter. Use this solution promptly after
capacity factor is not more than 2.5, the tailing factor is not preparation.
more than 1.4 and the relative standard deviation for replicate Reference solution (a). Weigh accurately a suitable quantity
injections is not more than 2.0 per cent. of ampicillin RS, dissolve in the solvent mixture by shaking
Inject alternately the test solution and reference solution (a). and mixing if necessary, with the aid of ultrasound to obtain a
Determine the weight per ml (2.4.29) of the suspension and
Use this solution promptly after preparation.
calculate the content of C16H19N3O4S, weight in volume.
Reference solution (b). Dissolve caffeine in reference solution
Repeat the procedure using a portion of the constituted
suspension that has been stored at the temperature and for
the period stated on the label. Chromatographic system
Labelling. The label states (1) the quantity of active ingredient – a stainless steel column 30 cm x 4.0 mm, packed with
in terms of the equivalent amount of ampicillin when the active octadecylsilane chemically bonded to porous silica or
ingredient is Ampicillin Trihydrate; (b) the temperature of ceramic microparticles (5 µm),
storage and the period during which the constituted – mobile phase: a mixture of 900 volumes of water,
suspension may be expected to be satisfactory for use. 80 volumes of acetonitrile, 10 volumes of 1 M
acid,
Ampicillin Dispersible Tablets – spectrophotometer set at 254 nm,
Dispersible Ampicillin Tablets – a 20 µl loop injector.
Ampicillin Dispersible Tablets contain Ampicillin or Ampicillin Inject reference solution (b). The resolution between the
Trihydrate in a suitable dispersible base. caffeine and ampicillin peaks is not less than 2.0. The relative
retention times are about 0.5 for ampicillin and 1.0 for caffeine.
Ampicillin Dispersible Tablets contain Ampicillin or Ampicillin
Trihydrate equivalent to not less than 90.0 per cent and not Inject reference solution (a). The test is not valid unless the
more than 120.0 per cent of the stated amount of ampicillin, capacity factor is not more than 2.5, the tailing factor is not
C16H19N3O4S. more than 1.4 and the relative standard deviation for replicate
Identification Inject alternately the test solution and reference solution (a).
In the Assay, the principal peak in the chromatogram obtained Calculate the percentage content of C16H19N3O4S in the tablets.
Storage. Store protected from moisture at a temperature not
exceeding 30°.
Tests Labelling. The label states (1) the strength in terms of the
Uniformity of dispersion. Place 2 tablets in 100 ml of water equivalent amount of ampicillin (when Ampicillin Trihydrate
and stir until completely dispersed. A smooth dispersion is is used); (2) that the tablets should be dispersed in water
produced, which passes through a sieve screen with a nominal immediately before use.
mesh aperture of 710 µm.
Ampicillin Trihydrate
Assay. Weigh and powder 20 tablets.
Determine by liquid chromatography (2. 4.14). H COOH
O
Solvent mixture. Mix 10 ml of 1 M monobasic potassium NH2 N CH3
H , 3H2O
phosphate and 1 ml of 1 M acetic acid and dilute to 1000 ml N CH3
S
with water. H H
O
Test solution. Transfer an accurately weighed quantity of the
powdered tablets containing about 100 mg of ampicillin to a
100-ml volumetric flask, add about 80 ml of the solvent mixture, C16H19N3O4S,3H2O Mol. Wt. 403.5
shake for 15 minutes and mix with the aid of ultrasound to Ampicillin Trihydrate is (6R)-6-(α-phenyl-D-glycyl-
achieve complete dissolution. Dilute to 100.0 ml with the amino)penicillanic acid trihydrate.
115
ALPHA AMYLASE IP 2007
Ampicillin Trihydrate contains not less than 96.0 per cent and Chromatographic system
not more than 100.5 per cent of C16H19N3O4S, calculated on the – a stainless steel column 30 cm x 4.0 mm, packed with
anhydrous basis. octadecylsilane chemically bonded to porous silica or
Description. A white, crystalline powder. ceramic microparticles (5 µm),
Identification 80 volumes of acetonitrile, 10 volumes of 1 M
A. Determine by infrared absorption spectrophotometry (2.4.6). acid,
Compare the spectrum with that obtained with ampicillin – flow rate. 2 ml per minute,
trihydrate RS or with the reference spectrum of ampicillin – spectrophotometer set at 254 nm,
trihydrate. – a 20 µl loop injector.
B. In the Assay, the principal peak in the chromatogram Inject reference solution (b). The resolution between the
obtained with the test solution corresponds to the peak in the caffeine and ampicillin peaks is not less than 2.0. The relative
chromatogram obtained with reference solution (a). retention times are about 0.5 for ampicillin and 1.0 for caffeine.
Tests Inject reference solution (a). The test is not valid unless the
capacity factor is not more than 2.5, the tailing factor is not
Appearance of solution. Dissolve 1.0 g in 10 ml of 1 M more than 1.4 and the relative standard deviation for replicate
hydrochloric acid and a further 1.0 g in a mixture of 3 ml of injections is not more than 2.0 per cent.
dilute ammonia solution and 7 ml of water. Both solutions
when freshly prepared are not more opalescent than
opalescence standard OS2 (2.4.1). Calculate the percentage content of C16H19N3O4S.
pH (2.4.24). 3.5 to 5.5, determined in a 0.25 per cent w/v solution. Storage. Store at a temperature not exceeding 30°.
Alpha Amylase
determined by Method B. Diastase
Heavy metals (2.3.13). 1.0 g complies with the limit test for Alpha Amylase is an amylolytic enzyme or a mixture of enzymes
heavy metals, Method B (20 ppm). obtained from fungi such as Aspergillus oryzae or from a
non-pathogenic variant of bacteria such as Bacillus subtilis
Sulphated ash (2.3.18). Not more than 0.5 per cent. and with the specific activity for converting starch into dextrin
Water (2.3.43). 12.0 per cent to 15.0 per cent, determined on and maltose. It may contain suitable harmless diluents such
0.1 g. as Lactose or Dibasic Calcium Phosphate.
Assay. Determine by liquid chromatography (2. 4.14). Alpha Amylase has amylase activity of not less than 800 Units
which represents the number of grams of dry, soluble maize or
corn starch digested by 1.0 g of Alpha Amylase under the
conditions of the Assay.
with water.
Description. A cream to light brown-coloured powder; almost
Test solution. Transfer an accurately weighed quantity odourless or with faint characteristic odour; hygroscopic.
containing about 100 mg of ampicillin to a 100-ml volumetric
flask and dissolve in the solvent mixture by shaking and mixing Tests
if necessary, with the aid of ultrasound and dilute to 100.0 ml
with the solvent mixture. Use this solution promptly after
on 1.0 g by drying in an oven at 105° for 1 hour.
preparation.
Assay. Weigh accurately a quantity containing 100 Units of
Reference solution (a). Weigh accurately a suitable quantity
amylase activity and triturate with 200 ml of buffer solution
pH 6.0 (for bacterial amylase) or of acetate buffer pH 5.0 (for
fungal amylase) and add sufficient buffer solution pH 6.0 or
acetate buffer pH 5.0, as appropriate, to produce 1000.0 ml.
Use this solution promptly after preparation.
Dilute 10.0 ml to 100.0 ml with buffer solution pH 6.0 or acetate
Reference solution (b). Dissolve caffeine in reference solution buffer pH 5.0, as appropriate, to give the test solution; filter if
(a) to obtain a solution containing about 0.12 mg per ml. necessary (1 ml of the test solution should be capable of
116
IP 2007 ANALGIN TABLETS
digesting about 10 mg of dry soluble maize or corn starch). Tests

Into each of six stoppered test-tubes add 5.0 ml of starch
substrate without touching the sides of the test-tube. Place Appearance of solution. A 5.0 per cent w/v solution in water is
the test-tubes in a water-bath maintained at 40° ± 0.1°. When clear (2.4.1).
the temperature of the solution in the tubes has reached 40°, Acidity or alkalinity. Dissolve 0.1 g in 10 ml of freshly boiled
add 0.35 ml, 0.4 ml, 0.45 ml, 0.5 ml, 0.55 ml and 0.6 ml of the test and cooled water and add a few drops of bromothymol blue
solution to each of the test-tubes marked 1 to 6 respectively solution. Not more than 0.05 ml of 0.01 M hydrochloric acid
and record the time of addition. Mix thoroughly and replace or 0.01 M sodium hydroxide is required to change the colour
the tubes in the water-bath. After exactly 60 minutes remove of the solution.
the tubes and cool rapidly in cold water. Add to each tube
Aminoantipyrine. Wet about 0.2 g with a few drops of water
0.05 ml of 0.02 M iodine and mix well. Note the tube containing
in a test-tube and add 3 ml of ethanol (95 per cent); shake
the lowest volume of test solution that does not show a bluish
until dissolved and add successively with shaking, 2 drops of
or violet tinge (if there is doubt, warm the solution slightly,
dilute ammonia solution, 5 drops of potassium ferricyanide
when the colour distinction is prominent). From this volume
solution, 2 drops of liquified phenol and 5 ml of water. The
calculate the number of grams of dry soluble maize or corn
solution acquires a green colour gradually but not an orange
starch digested by 1.0 g of the substance under examination.
or pink colour.
This represents the number of Units of amylase activity per g.
Arsenic (2.3.10). To 1.0 g in a long-necked, round-bottomed
flask add 5 ml of sulphuric acid and bring to the boil gently
and continue boiling on a low flame until the solution acquires
a light brown colour. Cool, add dropwise about 5 ml of
Analgin hydrogen peroxide solution (100 vol) and heat gently until
the solution just boils and continue the heating until the
Metamizol, Dipyrone
solution becomes colourless. Cool and add cautiously about
20 ml of water and mix. The resulting solution complies with
H3C the limit test for arsenic, but using 0.1 ml of arsenic standard
N solution (10 ppm As) and dipping the stained mercuric chloride
H3C N
, H2O papers from the test and standard solutions in a 10 per cent
w/v solution of potassium iodide before comparison of the
O
N stains (1 ppm).
Na O3S CH3 Heavy metals (2.3.13). Ignite 1.0 g until completely ashed.
Dissolve the residue in a mixture of 23 ml of water and 2 ml of
C13H16N3NaO4S,H2O Mol. Wt. 351.4 dilute acetic acid. The solution complies with the limit test
Analgin is sodium [N-(2,3-dihydro-1,5-dimethyl-3-oxo-2- for heavy metals, Method A (20 ppm).
phenyl-1H-pyrazol-4-yl)-N-methylamino]methanesulphonate Loss on drying (2.4.19). Not more than 5.5 per cent, determined
monohydrate. on 1.0 g by drying in an oven at 105°.
Analgin contains not less than 99.0 per cent and not more Assay. Weigh accurately about 0.4 g, dissolve in a mixture of
than 100.5 per cent of C13H16N3NaO4S, calculated on the dried 40 ml of ethanol (95 per cent) and 10 ml of 0.01 M
basis. hydrochloric acid and titrate with 0.05 M iodine until a yellow
Description. A white or almost white, crystalline powder with colour stable for 30 seconds is produced.
a scarcely perceptible yellowish tinge. 1 ml of 0.05 M iodine is equivalent to 0.01667 g of
C13H16N3NaO4S.
Identification
A. Wet about 0.1 g with two drops of water, add 5 ml of
ethanol (95 per cent) and 0.5 ml of dilute hydrochloric acid.
To the solution add 5 ml of potassium iodate solution; a Analgin Tablets
crimson colour is produced which deepens on further addition
of potassium iodate solution. Metamizol Tablets
B. Heat about 0.2 g with 2 ml of dilute hydrochloric acid; the Analgin Tablets contain not less than 95.0 per cent and not
characteristic odour of sulphur dioxide is produced followed more than 105.0 per cent of the stated amount of analgin,
by that of formaldehyde. C13H16N3NaO4S,H2O.
117
ANTICOAGULANT CITRATE DEXTROSE SOLUTION IP 2007
Identification NOTE — 15 ml of solution A or 25 ml of solution B are to be

used for 100 ml of whole blood.
Powder a few tablets and shake a quantity of the powder
containing about 0.5 g of Analgin with 10 ml of water and Description. A clear, colourless or faintly straw-coloured liquid;
filter. The filtrate complies with the following tests. odourless.
A. To 2 ml of the filtrate, add 5 ml of ethanol (95 per cent) and Identification

0.5 ml of dilute hydrochloric acid. To the solution add 5 ml of
potassium iodate solution; a crimson colour is produced A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
which deepens on further addition of potassium iodate the solution remains blue and clear. Heat to boiling, a copious
solution. red precipitate is formed.
B. Heat 4 ml of the filtrate with 2 ml of dilute hydrochloric B. Gives the reactions of sodium salts (2.3.1).
acid; the characteristic odour of sulphur dioxide is produced
C. To 2 ml (for Solution A) add 3 ml of water or to 4 ml (for
followed by that of formaldehyde.
Solution B) add 1 ml of water. The resulting solution gives
Tests reaction A of citrates (2.3.1).
Assay. Weigh and powder 20 tablets. Weigh accurately a pH (2.4.24). 4.5 to 5.5.
quantity of the powder containing about 0.5 g of Analgin and
transfer to a 50-ml volumetric flask. Add 10 ml of water and Bacterial endotoxins (2.2.3). Not more than 5.56 Endotoxin
shake for 1 minute. Dilute to volume with ethanol (95 per Units per ml.
cent), shake well and filter. Titrate 25.0 ml of the filtrate with Sterility (2.2.11). Complies with the test for sterility.
0.05 M iodine until a yellow colour stable for 30 seconds is
produced. Other tests. Complies with the tests stated under Parenteral
1 ml of 0.05 M iodine is equivalent to 0.01757 g of
C13H16N3NaO4S,H2O. Assay. For sodium citrate — Pipette 50.0 ml into a beaker and
titrate with 1.3 M hydrochloric acid to a pH of 1.98 ± 0.02,
Storage. Store protected from light and moisture. determining the end-point potentiometrically (2.4.25). Carry
out a blank titration with 50 ml of water.
1 ml of 1.3 M hydrochloric acid is equivalent to 0.1274 g of
Anticoagulant Citrate Dextrose C6H5Na3O7,2H2O.
Solution For free citric acid — Pipette 20.0 ml into a conical flask and
titrate with 0.1 M sodium hydroxide using phenolphthalein
ACD Solution solution as indicator.
Anticoagulant Citrate Dextrose Solution is a sterile solution 1 ml of 0.1 M sodium hydroxide is equivalent to 0.006404 g of
of Sodium Citrate, Citric Acid and Dextrose in Water for C6H8O7 or 0.007005 g of C6H8O7,H2O.
Injections.
For dextrose — Determine the optical rotation in a 2-dm tube
Anticoagulant Citrate Dextrose Solution contains not less than (2.4.22). The observed rotation multiplied by 1.0425, represents
95.0 per cent and not more than 105.0 per cent of the stated the weight of C6H12O6,H2O in 100 ml of the solution.
amounts of Sodium Citrate, C6H5Na3O7,2H2O, Citric Acid
monohydrate, C6H8O7,H2O (or Anhydrous Citric Acid, C6H8O7), Storage. Store protected from light in a single dose, tamper-
and Dextrose, C6H12O6,H2O. It contains no antimicrobial agent. evident container of colourless, transparent glass or of a
It is usually of two strengths as indicated below. suitable plastic material.
Solution A Solution B Labelling. The label states (1) whether the contents are
Sodium Citrate 2.20 g 1.32 g Solution A or Solution B; (2) volume of the solution required
per 100 ml of whole blood or the volume of the solution required
Citric Acid (Anhydrous) 0.73 g 0.44 g per volume of whole blood to be collected; (3) where applicable,
or Citric Acid (Monohydrate) 0.80 g 0.48 g the maximum amount of blood to be collected in the container.
Dextrose (Monohydrate) 2.45 g 1.47 g
Water for Injection 100 ml 100 ml
118
IP 2007 ANTICOAGULANT CITRATE PHOSPHATE DEXTROSE SOLUTION
Anticoagulant Citrate Phosphate 425 nm (2.4.7) using as the blank solution 1 ml of water treated
in the same manner. Prepare a calibration curve by measuring
Dextrose Solution the absorbance of solutions prepared by treating in the same
CPD Solution manner 1 ml quantities of suitable dilutions of a solution in
water containing 2.5 mg per ml of C6H8O7, prepared by using
Anticoagulant Citrate Phosphate Dextrose Solution is a sterile anhydrous citric acid, previously dried for 3 hours at 90°.
solution of Sodium Citrate, Citric Acid, Sodium Dihydrogen Calculate the total citrate content, as C6H8O7, in mg per ml of
Phosphate Dihydrate and Dextrose in Water for Injection. the solution under examination from the expression 0.2 C, where
Anticoagulant Citrate Phosphate Dextrose Solution contains C is the concentration in µg per ml of C6H8O7, read from the
not less than 95.0 per cent and not more than 105.0 per cent of curve.
the stated amounts of Sodium Citrate, C6H5Na3O7,2H2O, Citric Calculate the quantity, in mg, of C6H5Na3O7,2H2O in 1 ml of the
Acid, C6H8O7,H2O, Sodium Dihydrogen Phosphate Dihydrate, solution under examination from the expression 1.53 (A – B),
NaH2PO4,2H2O and Dextrose, C6H12O6,H2O. It contains no where A is the concentration in mg per ml of total citrate as
antimicrobial agent. It usually has the following composition: C6H8O7 and B is the concentration in mg per ml of free citric
Sodium Citrate 2.630 g acid in the solution.
Citric Acid (Monohydrate) 0.327 g For free citric acid — Pipette 20.0 ml into a conical flask and
Dextrose (Monohydrate) 2.550 g titrate with 0.1 M sodium hydroxide using phenolphthalein
Sodium Dihydrogen Phosphate
(Dihydrate) 0.251 g From the volume of 0.1 M sodium hydroxide required subtract
Water for Injection to 100 ml a volume, in ml, equal to 1.28 times the number of mg of
NaH2PO4,2H2O present, as determined in the Assay for sodium
NOTE – 14 ml are to be used for 100 ml of whole blood.
acid phosphate.
Description. A clear, colourless or faintly straw-coloured liquid;
1 ml of the remainder is equivalent to 0.007005 g of C6H8O7,H2O.
odourless.
For sodium dihydrogen phosphate dihydrate — Dilute 5.0 ml
Identification to 100.0 ml with water. Transfer 5.0 ml to a 25-ml graduated
flask and add 10.0 ml of a 2.8 per cent w/v solution of sulphuric
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution; acid followed by 2.0 ml of a 2.5 per cent w/v solution of
the solution remains blue and clear. Heat to boiling, a copious ammonium molybdate, mixing after each addition. Add 1.0 ml
red precipitate is formed. of aminohydroxynaphthalenesulphonic acid solution and
B. Gives the reactions of sodium salts and reaction B of sufficient water to produce 25.0 ml, mix and keep aside at 25°
phosphates (2.3.1). for 10 minutes. Measure the absorbance (A1) of the resulting
solution at the maximum at about 660 nm (2.4.7) using as the
C. To 2 ml add 3 ml of water. The resulting solution gives blank 5 ml of water treated in the same manner. Calculate the
reaction A of citrates (2.3.1). content of NaH2PO4,2H2O in each ml of the solution under
examination from the absorbance (A 2 ) obtained by
Tests simultaneously carrying out the operation using 5.0 ml of a
pH (2.4.24). 5.0 to 6.0. solution of potassium dihydrogen phosphate containing 0.11
mg of KH2PO4 per ml (C) and from the expression
Units per ml. 22.92 C (A1/A2).
Sterility (2.2.11). Complies with the test for sterility. For dextrose — Weigh a clean, medium- porosity sintered-
glass crucible containing a few glass beads. To 50 ml of
Other tests. Complies with the tests stated under Parenteral potassium cupri-tartrate solution add the glass beads from
Preparations (Injections). the weighed crucible, 45 ml of water and 5.0 ml of the solution
Assay. For sodium citrate — Dilute 25.0 ml to 100.0 ml with under examination. Heat the solution at such a rate that it
water and mix. Dilute 5.0 ml of the resulting solution to begins to boil in 3.5 to 4 minutes, boil the solution for exactly
100.0 ml with water and mix. Transfer 1.0 ml of this solution to 2 minutes and filter immediately through the weighed crucible,
a test-tube, add 1.3 ml of pyridine, swirl to mix, add 5.7 ml of taking care to transfer all the glass beads to the crucible, along
acetic anhydride, mix and immediately place in a water-bath with the precipitate. Wash the precipitate with hot water and
at 31° ± 0.5°. Allow the colour to develop for 35 minutes and then with 10 ml of ethanol (95 per cent) and dry it to constant
measure the absorbance of the resulting solution at about weight at 110°. Carry out a blank determination.
119
ANTICOAGULANT CITRATE PHOSPHATE DEXTROSE ADENINE SOLUTION IP 2007
1 mg of the precipitate is equivalent to 0.000496 g of D. In the test for adenine in the Assay, the principal peak in
C6H12O6,H2O. the chromatogram obtained with the test solution corresponds
Storage. Store in a single dose, tamper-evident container of to the peak in the chromatogram obtained with reference
colourless, transparent glass or of a suitable plastic material, solution (c).
Tests
Labelling. The label states (1) the composition and volume of
the solution; (2) volume of the solution required per 100 ml of pH (2.4.24). 5.0 to 6.0.
whole blood or the volume of the solution required per volume Bacterial endotoxins (2.2.3). Not more than 5.56 Endotoxin
of whole blood to be collected; (3) where applicable, the Units per ml.
maximum amount of blood to be collected in the container.
Assay. For total sodium — Dilute suitably with water and
Anticoagulant Citrate Phosphate determine by Method A for flame photometry (2.4.4), or by
Dextrose Adenine Solution Method A for atomic absorption spectrophotometry (2.4.3),
measuring at 589 nm and using sodium solution FP or sodium
CPDA Solution solution AAS respectively, suitably diluted with water for the
Anticoagulant Citrate Phosphate Dextrose Adenine Solution standard solutions.
is a sterile solution of Citric Acid, Sodium Citrate, Sodium For total citrate — Dilute 5.0 ml of the solution under
Dihydrogen Phosphate Dihydrate, Dextrose and Adenine in examination to 1000.0 ml with water and mix. Transfer 1.0 ml of
Water for Injection. this solution to a test-tube, add 1.3 ml of pyridine, swirl to mix,
Anticoagulant Citrate Phosphate Dextrose Adenine Solution add 5.7 ml of acetic anhydride, mix and immediately place in a
contains not less than 95.0 per cent and not more than 105.0 water-bath at 31° ± 1°. Allow the colour to develop for 33 ± 1
per cent of the stated amounts of total Sodium, Na, total Citrate, minutes and measure the absorbance of the resulting solution
C 6 H 5 O 7 , Sodium Dihydrogen Phosphate Dihydrate, at about 425 nm (2.4.7), using as the blank 1 ml of water treated
NaH2PO4,2H2O, Adenine,C5H5N5 and Dextrose Monohydrate, in the same manner. Prepare a calibration curve by measuring
C6H12O6,H2O. It contains no antimicrobial agent. It usually the absorbance of the solutions prepared by treating in the
has the following composition: same manner 1 ml quantities of suitable dilutions of a solution
in water containing 1.0 mg per ml of C6H8O7, prepared by
Citric Acid (Anhydrous) 0.2990 g using anhydrous citric acid, previously dried for 3 hours at
Sodium Citrate (Dihydrate) 2.6300 g 90°. Calculate the total citrate content, as C6H8O7, in mg per ml
Sodium Dihydrogen Phosphate of the solution under examination from the expression 0.2 C,
where C is the concentration in µg per ml of C6H8O7, read from
(Dihydrate) 0.2510 g
the curve.
Adenine 0.0275 g
For sodium dihydrogen phosphate dihydrate — Dilute 5.0 ml
Dextrose (Monohydrate) 3.1900 g
to 100.0 ml with water. Transfer 5.0 ml of this solution to a
Water for Injection to 100 ml 25-ml volumetric flask and add 10.0 ml of a 2.8 per cent w/v
NOTE — 14 ml are to be used for 100 ml of whole blood. solution of sulphuric acid followed by 2.0 ml of a 2.5 per cent
Description. A clear, colourless or faintly straw-coloured liquid; w/v solution of ammonium molybdate, mixing after each
odourless. addition. Add 1.0 ml of aminohydroxynaphthalenesulphonic
acid solution and sufficient water to produce 25.0 ml. Mix
Identification and keep aside at 25° for 10 minutes. Measure the absorbance
(A1) of the resulting solution at about 660 nm (2.4.7), using as
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution; the blank 5 ml of water treated in the same manner. Calculate
the solution remains blue and clear. Heat to boiling, a copious the content of NaH2PO4,2H2O in each ml of the solution under
red precipitate is formed. examination from the absorbance (A2) obtained by
B. Gives the reaction B of phosphates and the reactions of simultaneously carrying out the operation using 5.0 ml of a
sodium salts (2.3.1). solution of potassium dihydrogen phosphate containing
0.11 mg of KH2PO4 per ml (C) using the expression
C. To 2 ml add 3 ml of water. The resulting solution gives
reaction A of citrates (2.3.1). 25 C (A1/A2).
120
IP 2007 ARTEETHER
For adenine — Determine by liquid chromatography (2.4.14). Storage. Store protected from light, in single dose, tamper-
Test solution. Substance under examination. evident containers made of a suitable plastic material in a cool
place.
Reference solutions (a), (b) and (c) are prepared by dissolving
accurately weighed quantities of adenine RS in dilute Labelling. The label states (1) the composition and volume of
hydrochloric acid in three separate volumetric flasks, diluting the solution; (2) volume of the solution required per 100 ml of
with the same solvent to volume and mixing to obtain reference whole blood or the volume of the solution required per volume
solutions having known concentrations of about 0.25 mg, of whole blood to be collected; (3) where applicable, the
0.275 mg and 0.30 mg of adenine per ml respectively. maximum amount of blood to be collected in the container.
Reference solution (d). A solution containing 0.0275 per cent
w/v each of adenine RS and purine in dilute hydrochloric
acid. Arteether
– a stainless steel column 30 cm x 4 mm, packed with CH3
irregular or spherical, totally porous silica gel (10 µm) H
having a chemically bonded strongly acidic cation- OO
H3C
exchange coating, O
– mobile phase: dissolve 3.45 g of ammonium dihydrogen H
H H
phosphate in 950 ml of water in a 1000-ml volumetric O
flask, add 10 ml of glacial acetic acid, dilute to volume H CH3
with water and mix, O CH3
– spectrophotometer set at 254 nm, C17H28O5 Mol. Wt. 312.4
Arteether is dihydroartemisinin ethyl ether.
Inject solution (d) at least four times and record the
chromatograms. The test is not valid unless the relative Arteether contains β-isomer not less than 25.0 per cent and
standard deviation of the peak response of adenine is not not more than 35.0 per cent and β-isomer not less than 65.0 per
more than 2.5 per cent, the relative standard deviation of the cent and not more than 75.0 per cent and total arteether is not
retention time of adenine is not more than 2.0 per cent and the less than 95.0 per cent and not more than 105.0 per cent of
resolution factor of adenine and purine is not less than 3.0. C17H28O5, calculated on the dried basis.
Inject separately the test solution and reference solutions (a), Description. A light yellow coloured semi-solid, lipophylic
(b) and (c). Record the chromatograms and measure the powder.
responses for the major peaks. Plot the responses against the
Identification
concentrations in mg of adenine per ml of reference solutions
(a), (b) and (c). A. Determine by infrared absorption spectrophotometry (2.4.6).
Calculate the quantity, in mg, of C5H5N5 in each ml of the Compare the spectrum with that
solution under examination as the value read directly from the obtained with arteether RS or with the reference spectrum of
standard curve corresponding to the response obtained with arteether.
the test solution.
For dextrose — Weigh a clean, medium porosity sintered- obtained with test solution corresponds to the
glass crucible containing a few glass beads. To 50 ml of
potassium cupri-tartrate solution add the glass beads from peak obtained in the chromatogram with reference solution.
the weighed crucible, 45 ml of water and 5.0 ml of the solution C. Determine by thin layer chromatography (2.4.17), coating
under examination. Heat the solution at such a rate that it the plate with silica gel GF254.
begins to boil in 3.5 to 4 minutes, boil the solution for exactly
Mobile phase. A mixture of 10 volumes of ethyl acetate and 90
2 minutes and filter immediately through the weighed crucible,
volumes of hexane.
taking care to transfer all the glass beads with the precipitate
to the crucible. Wash the precipitate with hot water and then Test solution. Dissolve 20 mg of the substance under
with 10 ml of ethanol (95 per cent) and dry it to constant examination in 10 ml of ethyl acetate.
weight at 110°. Carry out a blank determination. Reference solution. A solution containing 0.2 per cent w/v
1 mg of the precipitate is equivalent to 0.000496 g of C6H12O6. each of á-arteether RS and ß–arteether RS in ethyl acetate.
121
ARTEMETHER IP 2007
Apply to the plate 5 µl of each solution. Allow the mobile – spectrophotometer set at 235 nm,
phase to rise 8 cm. Dry the plate in air, spray with a mixture of – a 20 µl loop injector.
50 volumes of glacial acetic acid, 1 volume of sulphuric acid Inject the reference solution. The test is not valid unless the
and 0.5 volume of anisaldehyde and heat at 100° for 15 minutes. tailing factor is not more than 1.5 for both component. The
The principal spots in the chromatogram obtained with the relative standard deviation for replicate injections is not more
test solution corresponds to that in the chromatogram obtained than 2.0 per cent.
with reference solution. Inject the test solution and the reference solution.
Tests Calculate the content of C17H28O5.
Appearance of solution. A 40.0 per cent w/v solution in Storage. Store protected from light and moisture.
methanol is clear (2.4.1).
Specific optical rotation (2.4.22). +92.9º to +93.7º, determined
in a 1.8 per cent w/v solution in chloroform.
Artemether
(2.4.14).
CH3
Test solution. Dissolve 100 mg of the substance under H
examination in 100 ml of acetonitrile. O O
H3C
Reference solution (a). A solution containing 0.1 per cent w/ O
H
v each of á –arteether RS and ß –arteether RS in acetonoitrile. H H
O
Reference solution (b). Dilute 1 ml of reference solution (a) to H CH3
100 ml with acetonoitrile. OCH3
Chromatographic system as described under Assay.
C16H26O5 Mol. Wt. 298.4
column efficiency is not less than 2000 theoretical plates and Arteether is dihydroartemisinin methyl ether.
the tailing factor is not more than 2.0 for both component. Artemether contains not less than 98.0 per cent and not more
Inject the test solution and reference solution (b). In the than 102.0 per cent of C16H26O5, calculated on the dried basis.
chromatogram obtained with the test solution, the area of any Description. A white crystalline, odourless, slightly bitter,
secondary peak is not more than 0.5 times the area of the lipophylic substance.
peaks in the chromatogram obtained with the reference
solution (b) (0.5 per cent) and the sum of areas of all the Identification
secondary peaks is not more than twice the area of the peaks
in the chromatogram obtained with the reference solution (b) A. Determine by infrared absorption spectrophotometry (2.4.6).
(2.0 per cent). Compare the spectrum with that obtained with artemether RS
or with the reference spectrum of artemether.
on 1.0 g by drying in an oven at 80º.
C. Determine by thin layer chromatography (2.4.17), coating
Test solution. Dissolve 100 mg of the substance under the plate with silica gel GF254.
examination in 100 ml of acetonitrile.
Mobile phase. A mixture of 10 volumes of ethyl acetate and
Reference solution. A solution containing 0.1 per cent w/v 90 volumes of hexane.
each of β –arteether RS and β –arteether RS in acetonoitrile.
Chromatographic system examination in 10 ml of ethyl acetate.
Reference solution. A 0.2 per cent w/v solution of artemether
RS in ethyl acetate.
32 volumes of water and 3 volumes of 1,4-dioxane Apply to the plate 5 µl of each solution. Allow the mobile
– flow rate.1.2 ml per minute, phase to rise 8 cm. Dry the plate in air, spray with a mixture of
122
IP 2007 ARTEMISININ
50 volumes of glacial acetic acid, 1 volume of sulphuric acid Chromatographic system

and 0.5 volume of anisaldehyde and heat at 100° for 15 minutes. – a stainless steel column 25 cm x 4.0 mm, packed with
The principal spot in the chromatogram obtained with the test octadecylsilane bonded to porous silica (5 µm),
solution corresponds to that in the chromatogram obtained – mobile phase: a mixture of 65 volumes of acetonitrile,
with reference solution. 32 volumes of water and 3 volumes of 1,4-dioxane,
D. Dissolve 5 mg in 1 ml of ethanol anhydrous and add 20 mg – flow rate.1.2 ml per minute,
of potassium iodide. Heat the mixture on a water-bath. A yellow – spectrophotometer set at 235 nm,
colour is produced. – a 20 µl loop injector.
E. Dissolve 5 mg in 1 ml of ethanol anhydrous. Add a few
tailing factor is not more than 1.5. The relative standard
drops on a white porcelain dish and add 1 drop
deviation for replicate injections is not more than 2.0 per cent.
of vanillin sulphuric acid TS. A pink colour is produced. Inject the test solution and the reference solution.
Appearance of solution. Dissolve 0.5 g in 0.5 ml of methanol, Storage. Store protected from light and moisture.
this solution is clear (2.4.1) and colourless (2.4.1).
Specific optical rotation (2.4.22). +159.30 to +160.20 at 34º,
determined in a 1.0 per cent w/v solution in chloroform.
Artemisinin
Related substances. Determine by liquid chromatography CH3
(2.4.14). H
Test solution. Dissolve 100 mg of the substance under OO
H3C
examination in 100 ml of acetonitrile. O
H
Reference solution (a). A 0.1 per cent w/v solution of H H
O
artemether RS in acetonoitrile. CH3
Reference solution (b). Dilute 1 ml of reference solution (a) to O
100 ml with acetonoitrile. C15H22O5 Mol.Wt. 282.3
Chromatographic system as described under Assay. Artemesine is (3R,5aS,6R,8aS,12S,12aR)-octahydro-3,6,9-
Inject reference solution (a). The test is not valid unless the trimethyl-3,12-epoxy-12H-pyrano[4,3-j]-1,2-benzodioxepin-
column efficiency is not less than 2000 theoretical plates and 10(3H)-one.
the tailing factor is not more than 2.0. Artemisinin contains not less than 98.0 per cent and not more
Inject the test solution and reference solution (b). In the than 103.0 per cent of artemisinin, C15H22O5, calculated on the
chromatogram obtained with the test solution, the area of any dried basis.
secondary peak is not more than 0.5 times the area of the peak Description. A white crystalline powder.
in the chromatogram obtained with the reference solution (b)
(0.5 per cent) and the sum of areas of all the secondary peaks Identification
is not more than twice the area of the peak in the chromatogram
obtained with the reference solution (b)
Compare the spectrum with that obtained with artemisinin RS
(2.0 per cent).
or with the reference spectrum of artemisinin.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined obtained with the test solution corresponds to the peak in the
on 1.0 g by drying over phosphorous pentaoxide under chromatogram obtained with the reference solution.
vacuum at 2.67 kPa.
C. Determine by thin layer chromatography (2.4.17), coating
Assay. Determine by liquid chromatography (2.4.14). the plate with silica gel GF254.
Test solution. Dissolve 100 mg of the substance under Mobile phase. A mixture of 15 volumes of ethyl acetate and 85
examination in 100.0 ml in acetonitrile. volumes of hexane.
Reference solution. A 0.1 per cent w/v solution of artemether Test solution. Dissolve 2 mg of the substance under
RS in acetonitrile. examination in 1 ml of ethyl acetate.
123
ASCORBIC ACID IP 2007
Reference solution. A 0.2 per cent w/v solution of artemisinin – mobile phase: a mixture of 65 volumes of acetonitrile,
RS in ethyl acetate. 32 volumes of water and 3 volumes of 1,4-dioxane.
Apply to the plate 5 µl of each solution as bands 10 mm by 2 – flow rate.1.2 ml per minute,
mm. Allow the mobile phase to rise 8 cm. Dry the plate in air, – spectrophotometer set at 235 nm,
spray with a mixture of 50 volumes of glacial acetic acid, 1 – a 20 µl loop injector.
volume of sulphuric acid and 0.5 volume of anisaldehyde Inject the reference solution. The test is not valid unless the
and heat at 100° for 15 minutes. The chromatogram obtained tailing factor is not more than 1.5. The relative standard
with test solution shows pink band corresponding to the band deviation for replicate injections is not more than 2.0 per cent.
in the chromatogram obtained with reference solution. Inject the test solution and the reference solution.
Appearance of solution. Dissolve 0.5 g in 0.5 ml of chloroform,
the solution is clear (2.4.1) and colourless (2.4.1).
Specific optical rotation (2.4.22). +64.70 to +65.40, determined Ascorbic Acid
Vitamin C; L-Ascorbic Acid
(2.4.14). CH2OH
Test solution. Dissolve 100 mg of the substance under H COH
examination in 100 ml of acetonitrile. O O
artemisinin RS in acetonoitrile. HO OH
C6H8O6 Mol. Wt. 176.1
100 ml with acetonoitrile.
Ascorbic Acid is (R)-5-[(S)-1,2-dihydroxyethyl)-3,4-dihydroxy-
Chromatographic system as described in the Assay.
5(H)-furan-2-one.
Ascorbic Acid contains not less than 99.0 per cent and not
column efficiency is not less than 2000 theoretical plates and
more than 100.5 per cent of C6H8O6.
Description. A white to very pale yellow crystalline powder or
colourless crystals; odourless. On exposure to light it gradually
darkens.
in the chromatogram obtained with the reference solution (b) Identification
is not more than twice the area of the peak in the chromatogram Test A may be omitted if tests B, C and D are carried out. Tests
obtained with the reference solution (b) C and D may be omitted if tests A and B are carried out.
(2.0 per cent). A. Determine by infrared absorption spectrophotometry (2.4.6).
Sulphated ash (2.3.18). Not more than 0.1 per cent w/w. Compare the spectrum with that obtained with ascorbic acid
RS or with the reference spectrum of ascorbic acid.
on 1.0 g by drying in an oven at 80º. B. Add 2 ml of a 2 per cent w/v solution to a few ml of 2,6-
dichlorophenolindophenol solution; the solution is
Assay. Determine liquid chromatography (2.4.14). decolorised.
Test solution. Dissolve 100 mg of the substance under C. Dilute 1 ml of a 2 per cent w/v solution with 5 ml of water
examination in 100 ml of acetonitrile. and add 1 drop of a freshly prepared 5 per cent w/v solution of
Reference solution. A 0.1 per cent w/v solution of artemisinin sodium nitroprusside and 2 ml of dilute sodium hydroxide
RS in acetonoitrile. solution. Add 0.6 ml of hydrochloric acid dropwise and stir;
the yellow colour turns blue.
– a stainless steel column 25 cm x 4.0 mm, packed with D. To 2 ml of a 2 per cent w/v solution add 2 ml of water, 0.1 g
octadecylsilane bonded to porous silica (5 µm), of sodium bicarbonate and about 20 mg of ferrous sulphate,
124
IP 2007 ASCORBIC ACID TABLETS
shake and allow to stand; a deep violet colour is produced. solution followed immediately by 1 ml of 0.1 M sodium
Add 5 ml of 1 M sulphuric acid; the colour disappears. hydroxide; a transient blue colour is produced.
Tests B. To a volume containing 40 mg of Ascorbic Acid add 4 ml of

0.1 M hydrochloric acid and 4 drops of methylene blue
Appearance of solution. A 5.0 per cent w/v solution in water is solution and warm to 40°; the deep blue colour becomes
clear (2.4.1), and not more intensely coloured than reference appreciably lighter or is completely discharged within
solution BYS7 (2.4.1). 3 minutes.
pH (2.4.24). 2.2 to 2.5, determined in a 5.0 per cent w/v solution. C. The solution responds to the flame test for sodium salts
Specific optical rotation (2.4.22). +20.5° to +21.5°, determined (2.3.1).
Tests
Light absorption. Absorbance (2.4.7) of a 0.001 per cent w/v
solution in 0.01 M hydrochloric acid at the maximum at about pH (2.4.24). 5.5 to 7.0.
244 nm, about 0.56.
Oxalic acid. Dilute a volume containing 0.25 g of Ascorbic
Oxalic acid. Dissolve 0.25 g in 5 ml of water and neutralise to Acid in 5 ml of water and neutralise to litmus paper with 2 M
litmus paper with 2 M sodium hydroxide. Add 1 ml of 2 M sodium hydroxide. Add 1 ml of 2 M acetic acid and 0.5 ml of
acetic acid and 0.5 ml of 0.5 M calcium chloride. Any 0.5 M calcium chloride. Any opalescence, after 60 minutes, is
opalescence, after 60 minutes, is not more intense than that not more intense than that produced by treating 5 ml of a
produced by treating 5 ml of a solution prepared by dissolving solution prepared by dissolving 70 mg of oxalic acid in 500 ml
70 mg of oxalic acid in 500 ml of water in a similar manner (0.3 of water in a similar manner (0.3 per cent).
per cent).
Heavy metals (2.3.13). 1.0 g dissolved in 25 ml of water complies Preparations (Injections).
with the limit test for heavy metals, Method A (20 ppm).
Assay. Measure accurately a volume containing about 50 mg
Sulphated ash (2.3.18). Not more than 0.1 per cent. of Ascorbic Acid and transfer to a 250-ml volumetric flask.
Assay. Weigh accurately about 0.1 g and dissolve in a mixture Add 20 ml of metaphosphoric-acetic acids solution, dilute
of 100 ml of freshly boiled and cooled water and 25 ml of 1 M with water to 250.0 ml and mix. Pipette 10.0 ml into a 50-ml
sulphuric acid. Immediately titrate with 0.05 M iodine, using Erlenmeyer flask, add 5 ml of metaphosphoric-acetic acids
starch solution as indicator until a persistent blue-violet colour solution and titrate with standard 2,6-dichlorophenolindo-
is obtained. phenol solution, until the pink colour persists for at least 10
1 ml of 0.05 M iodine is equivalent to 0.008806 g of C6H8O6. seconds, the titration occupying not more than 2 minutes.
Repeat the operation with a mixture of 5.5 ml of
Storage. Store protected from light and moisture avoiding metaphosphoric-acetic acids solution and 15 ml of water
contact with metals. It undergoes rapid decomposition in omitting the preparation being examined. From the difference
solutions in contact with air. calculate the ascorbic acid in each ml of the injection from the
ascorbic acid equivalent of the standard 2,6-
dichlorophenolindophenol solution.
Ascorbic Acid Injection
Storage. Store protected from light, in a single dose container.
Vitamin C Injection; L-Ascorbic Acid Injection
Ascorbic Acid Injection is a sterile solution of Sodium
Ascorbate or of Ascorbic Acid prepared with the aid of Sodium Ascorbic Acid Tablets
Hydroxide or Sodium Carbonate or Sodium Bicarbonate in
Water for Injections. Vitamin C Tablets; L-Ascorbic Acid Tablets
Ascorbic Acid Injection contains not less than 95.0 per cent Ascorbic Acid Tablets contain not less than 95.0 per cent and
and not more than 115.0 per cent of the stated amount of not more than 115.0 per cent of the stated amount of ascorbic
ascorbic acid, C6H8O6. acid, C6H8O6. The tablets may contain permitted flavouring
Description. A clear, colourless liquid. agents.
Identification Identification
A. To a volume containing 5 mg of Ascorbic Acid add 0.5 ml of A. Shake a quantity of the powdered tablets with sufficient
0.1 M hydrochloric acid and 3 drops of sodium nitroprusside water to make approximately the equivalent of a 2 per cent
125
ASPARTAME IP 2007
w/v solution of Ascorbic Acid and filter. The filtrate (solution B. When examined in the range 230 nm to 300 nm (2.4.7), a
A) is acid to litmus solution. 0.1 per cent w/v solution in ethanol (95 per cent) shows
B. To solution A add a few ml of 2,6-dichlorophenolindo- absorption maxima at about 247 nm, 252 nm, 258 nm and
phenol solution; the solution is decolorised. 264 nm.
C. To 1 ml of solution A, add about 0.1 ml of 2 M nitric acid Tests

and 0.05 ml of silver nitrate solution; a grey precipitate is
produced. pH (2.4.24). About 5.0, determined in a 0.8 per cent w/v solution.
Specific optical rotation (2.4.22). +14.5° to +16.5°, determined
Tests at 20° in a 4.0 per cent w/v solution in 15 M formic acid within
Disintegration. The test does not apply to Ascorbic Acid 30 minutes of preparing the solution.
Tablets containing 500 mg or more of Ascorbic Acid. Light absorption (2.4.7). Absorbance of a 1.0 per cent w/v
solution in 2 M hydrochloric acid, prepared with the aid of
ultrasound, at the maximum at about 430 nm, not more than
Assay. Weigh and powder 20 tablets. Weigh accurately a 0.022.
quantity of the powder containing about 0.15 g of Ascorbic
5-Benzyl-3,6-dioxo-2-piperazineacetic acid. Determine by
Acid and dissolve as completely as possible in a mixture of
liquid chromatography (2.4.14).
30 ml of water and 20 ml of 1 M sulphuric acid. Titrate with
0.1 M ceric ammonium sulphate using ferroin sulphate Test solution. Dissolve 0.5 g of the substance under
solution as indicator. examination in 100 ml of a mixture of 10 volumes of methanol
1 ml of 0.1 M ceric ammonium sulphate is equivalent to and 90 volumes of water.
0.008806 g of C6H8O6. Reference solution. A 0.0075 per cent w/v solution of 5-benzyl-
3,6-dioxo-2-piperazineacetic acid RS in a mixture of 10
Storage. Store protected from light and moisture avoiding
volumes of methanol and 90 volumes of water.
contact with metals.
Labelling. For tablets containing 500 mg or more of Ascorbic
Acid the label states, where applicable, that the tablets should
be chewed before swallowing.
ceramic microparticles (3 to 10 µm),
– mobile phase: dissolve 5.6 g of potassium dihydrogen
phosphate in 820 ml of water, adjust to pH 4.3 with
Aspartame phosphoric acid and dilute to 1000 ml with methanol,
O H H NH2 – a 20 µl loop injector.
H3 C N COOH
O Inject the test solution and reference solution. Record the
H O chromatograms. The test is not valid if the relative standard
deviations for replicate injections is more than 4.0 per cent
and the symmetry factor of the principle peak in the
chromatogram obtained with the reference solution is more
than 2.0.
C14H18N2O5 Mol. Wt. 294.3
In the chromatogram obtained with the test solution the
Aspartame is N-L-α -aspartyl-L-phenylalanine 1-methyl ester. response obtained for any peak at a retention time
Aspartame contains not less than 98.0 per cent and not more corresponding to that of 5-benzyl-3,6-dioxo-2-
than 102.0 per cent of C14H18N2O5, calculated on the dried piperazineacetic acid RS is not greater than the response
basis. obtained for the peak in the chromatogram of the reference
solution corresponding to not more than 1.5 per cent of 5-
Description. A white, crystalline powder; odourless. benzyl-3,6-dioxo-2-piperazineacetic acid.
Identification Other Related substances. Carry out the test for 5-Benzyl-
3,6-dioxo-2-piperazineacetic acid, using reference solution (b)
A. Determine by infrared absorption spectrophotometry (2.4.6). prepared by diluting 2.0 ml of the test solution to 100 ml with
Compare the spectrum with that obtained with aspartame RS. a mixture of 10 volumes of methanol and 90 volumes of water.
126
IP 2007 ASPIRIN
Inject 20 µl of the test solution and reference solution (b), Identification

record the chromatograms and measure the peak responses.
Continue elution of the test solution for twice the retention Test A may be omitted if tests B and C are carried out. Tests B
time of the aspartame peak. The sum of the areas of any peaks and C may be omitted if test A is carried out.
observed in the chromatogram obtained with the test solution, A. Determine by infrared absorption spectrophotometry (2.4.6)
other than the peaks for aspartame and 5-benzyl-3,6-dioxo-2- Compare the spectrum with that obtained with aspirin RS or
piperazineacetic acid, is not more than the area of the with the reference spectrum of aspirin.
aspartame peak obtained with reference solution (b)(2.0 per
B. Boil about 0.5 g with 10 ml of sodium hydroxide solution
cent).
for 3 minutes, cool and add 10 ml of dilute sulphuric acid; a
Arsenic (2.3.10). Mix 3.3 g with 3 g of anhydrous sodium white, crystalline precipitate is produced and the odour of
carbonate, add 10 ml of bromine solution and mix thoroughly. acetic acid is perceptible. Filter, dissolve the precipitate in
Evaporate to dryness on a water-bath, gently ignite, dissolve about 2 ml of water and add ferric chloride test solution; a
the cooled residue in 16 ml of brominated hydrochloric acid deep violet colour is produced.
AsT and add 45 ml of water. Remove the excess of bromine
C. To the filtrate obtained in test B add 3 ml of ethanol (95 per
with 2 ml of stannous chloride AsT. The resulting solution
cent) and 3 ml of sulphuric acid and warm; the odour of ethyl
acetate is perceptible.
heavy metals, Method B (10 ppm). Tests
Sulphated ash (2.3.18). Not more than 0.2 per cent, determined Appearance of solution. A 1.0 per cent w/v solution in ethanol
on 2.0 g. (95 per cent) is clear (2.4.1) and not more intensely coloured
than reference solution BS8 (2.4.1).
on 1.0 g by drying in an oven at 105° for 4 hours. Clarity of solution in alkali. A 5.0 per cent w/v solution in a
warm 5 per cent w/v solution of sodium carbonate is clear
Assay. Weigh accurately about 0.3 g, dissolve in 1.5 ml of
(2.4.1).
anhydrous formic acid, add 60 ml of anhydrous glacial acetic
acid. Titrate with 0.1 M perchloric acid, using crystal violet Arsenic (2.3.10). Mix 5.0 g with 3 g of anhydrous sodium
solution as indicator. Carry out a blank titration. A blank carbonate, add 10 ml of bromine solution and mix thoroughly.
titration of more than 0.1 ml may be indicative of excessive Evaporate to dryness on a water-bath, gently ignite, and
water content. In such a case, repeat the test after taking dissolve the cooled residue in 16 ml of brominated
precautions to maintain anhydrous conditions throughout. hydrochloric acid and 45 ml of water. Remove the excess of
bromine with 2 ml of stannous chloride AsT. The resulting
C14H18N2O5.
Heavy metals. Not more than 10 ppm, determined by the
following method. Dissolve 2.0 g in 25 ml of acetone, add 1 ml
of water and 10 ml of hydrogen sulphide solution; any colour
produced is not more intense than that produced by mixing 25
Aspirin ml of acetone, 1.0 ml of lead standard solution (20 ppm Pb)
and 10 ml of hydrogen sulphide solution.
Acetylsalicylic Acid
Chlorides (2.3.12). Boil 1.75 g with 75 ml of water for 5 minutes,
COOH cool, add sufficient water to restore the original volume and
O CH3 filter. 25 ml of the filtrate complies with the limit test for chlorides
(430 ppm).
O Sulphates (2.3.17). 10 ml of the filtrate obtained in the test for
Chlorides complies with the limit test for sulphates (600 ppm).
C9H8O4 Mol. Wt. 180.2
Readily carbonisable substances. Dissolve 0.5 g in 5 ml of
Aspirin is 2-acetoxybenzoic acid. sulphuric acid (containing 94.5 per cent to 95.5 per cent w/w
Aspirin contains not less than 99.5 per cent and not more than of H2SO4); any colour produced is not more intense than that
100.5 per cent of C9H8O4, calculated on the dried basis. of reference solution BYS4 (2.4.1).
Description. Colourless crystals or a white, crystalline powder; Salicylic acid. Dissolve 2.5 g in sufficient ethanol (95 per
odourless or almost odourless. cent) to produce 25.0 ml (test solution). To each of two matched
127
ASPIRIN TABLETS IP 2007
Nessler cylinders add 48 ml of water and 1 ml of a freshly salicylic acid, 2 ml of ethanol (95 per cent) and sufficient
prepared acid ferric ammonium sulphate solution. Into one water to produce 50 ml contained in a second Nessler cylinder
cylinder add 1.0 ml of a freshly prepared 0.01 per cent w/v (0.3 per cent).
solution of salicylic acid and into the other pipette 1.0 ml of Other tests. Comply with the tests stated under Tablets.
the test solution. Mix the contents of the cylinders; after
30 seconds, the colour in the cylinder containing the test Assay. Weigh and powder 20 tablets. Weigh accurately a
solution is not more intense than that in the cylinder containing quantity of the powder containing about 0.5 g of Aspirin, add
the standard solution (0.1 per cent). 30.0 ml of 0.5 M sodium hydroxide, boil gently for 10 minutes,
cool and titrate the excess of alkali with 0.5 M hydrochloric
acid using phenol red solution as indicator. Repeat the
Loss on drying (2.4.19). Not more than 0.5 per cent, determined operation without the substance under examination. The
on 1.0 g by drying over phosphorus pentoxide at a pressure difference between the titrations represents the amount of
of 1.5 to 2.5 kPa. sodium hydroxide required.
Assay. Weigh accurately about 1.5 g, dissolve in 15 ml of 1 ml of 0.5 M sodium hydroxide is equivalent to 0.04504 g of
ethanol (95 per cent), add 50.0 ml of 0.5 M sodium hydroxide, C9H8O4.
boil gently for 10 minutes, cool and titrate the excess of alkali Storage. Store protected from moisture at a temperature not
with 0.5 M hydrochloric acid using phenol red solution as exceeding 30°.
indicator. Repeat the operation without the substance under
examination. The difference between the titrations represents
the amount of sodium hydroxide required. Soluble Aspirin Tablets
C9H8O4.
Dispersible Aspirin Tablets; Calcium Aspirin Tablets
Soluble Aspirin Tablets contain not less than 90.0 per cent
and not more than 110.0 per cent of the stated amount of
exceeding 30°.
aspirin, C9H8O4.
Identification
Aspirin Tablets
A. The tablets effervesce on the addition of water.
Acetylsalicylic Acid Tablets
B. Boil 0.1 g of the powdered tablets with 10 ml of water and
Aspirin Tablets contain not less than 95.0 per cent and not 0.5 ml of ferric chloride test solution; a violet-red colour is
more than 105.0 per cent of the stated amount of aspirin, C9H8O4. produced.
Identification Tests
Boil a quantity of the powdered tablets containing 0.5 g of Salicylic acid. To a quantity of the powdered tablets containing
Aspirin with 10 ml of sodium hydroxide solution for 3 minutes, 0.5 g of Aspirin add 25.0 ml of chloroform, shake vigorously
cool and add 10 ml of dilute sulphuric acid; a white, crystalline for 2 minutes and filter through a dry filter paper. Evaporate
precipitate is produced and the odour of acetic acid is 5.0 ml of the filtrate rapidly to dryness in a dish in a current of
perceptible. Filter, dissolve the precipitate in about 2 ml of dry air at room temperature. Dissolve the residue in 2 ml of
water and add ferric chloride test solution; a deep violet ethanol (95 per cent), transfer to a Nessler cylinder, using a
colour is produced. further 1 ml of ethanol (95 per cent) to rinse the dish, dilute to
50 ml with water, add 1 ml of acid ferric ammonium sulphate
Tests solution, mix, and allow to stand for 1 minute; the violet colour
produced is not more intense than that produced by adding 1
Salicylic acid. Shake a quantity of the powdered tablets ml of acid ferric ammonium sulphate solution to a mixture of
containing 0.2 g of Aspirin with 4 ml of ethanol (95 per cent), 2.0 ml of a freshly prepared 0.15 per cent w/v solution of
dilute to 100.0 ml with water, filter immediately, transfer 50 ml salicylic acid, 3 ml of ethanol (95 per cent) and sufficient
of the filtrate to a Nessler cylinder, add 1.0 ml of freshly water to produce 50 ml contained in a second Nessler cylinder
prepared acid ferric ammonium sulphate solution, mix and (3 per cent).
allow to stand for 1 minute; the violet colour produced is not
more intense than that produced by adding 1 ml of freshly Other tests. Comply with the tests stated under Tablets.
prepared acid ferric ammonium sulphate solution to a mixture Assay. Weigh and powder 20 tablets. Weigh accurately a
of 3.0 ml of a freshly prepared 0.01 per cent w/v solution of quantity of the powder containing about 0.3 g of Aspirin,
128
IP 2007 ATENOLOL
dissolve in 10 ml of 0.5 M sulphuric acid and boil under a ml of water and allow to separate. Filter the chloroform layer
reflux condenser for 1 hour. Cool, transfer to a separating through a dry filter paper and evaporate 10 ml of the filtrate to
funnel with the aid of small quantities of water, and extract the dryness at room temperature using a rotary evaporator. To the
liberated salicylic acid with four quantities, each of 20 ml, of residue add 4 ml of ethanol (95 per cent), stir well, dilute to
ether. Wash the combined ether extracts with two quantities, 100 ml with water at a temperature not exceeding 10°, filter
each of 5 ml, of water, remove the ether in a current of air at a immediately, rapidly transfer 50 ml to a Nessler cylinder, add 1
temperature not exceeding 30°, dissolve the residue in 20 ml of ml of freshly prepared acid ferric ammonium sulphate
0.5 M sodium hydroxide, and dilute to 200.0 ml with water. solution, mix and allow to stand for 1 minute; the violet colour
Transfer 50.0 ml to a stoppered flask, add 50.0 ml of 0.05 M produced is not more intense than that produced by adding 1
bromine and 5 ml of hydrochloric acid, protect the mixture ml of acid S ammonium sulphate solution to a mixture of 3.0 ml
from light and shake repeatedly during 25 minutes. Add 20 ml of a freshly prepared 0.01 per cent w/v solution of salicylic
of potassium iodide solution, shake thoroughly and titrate acid, 2 ml of ethanol (95 per cent) and sufficient water to
with 0.1 M sodium thiosulphate using starch solution, added produce 50 ml contained in a second Nessler cylinder (0.6 per
towards the end of the titration, as indicator. cent).
1 ml of 0.05 bromine is equivalent to 0.003003 g of C9H8O4. Other tests. Comply with the tests stated under Tablets.
Storage. Store protected from moisture at a temperature not Assay. Weigh and powder 20 tablets.
exceeding 30°. For aspirin — Weigh accurately a quantity of the powder
Labelling. The label states that the tablets should be dispersed containing about 0.7 g of Aspirin, add 20 ml of water and 2 g
in water immediately before use of sodium citrate and heat under a reflux condenser for
30 minutes. Cool, wash the condenser with 30 ml of warm
water and titrate with 0.5 M sodium hydroxide using
Aspirin And Caffeine Tablets phenolphthalein solution as indicator.
Acetylsalicylic Acid and Caffeine Tablets 1 ml of 0.5 M sodium hydroxide is equivalent to 0.04504 g of
C9H8O4.
Aspirin and Caffeine Tablets contain not less than 330 mg and
not more than 370 mg of aspirin, C9H8O4, and not less than For caffeine — Weigh accurately a quantity of the powder
27.5 mg and not more 32.5 mg of caffeine, C8H10N4O2. containing about 30 mg of Caffeine add 200 ml of water and
shake for 30 minutes. Add sufficient water to produce 250.0
Identification ml and filter. To 10.0 ml of the filtrate add 10 ml of 1 M sodium
hydroxide and extract immediately with five quantities, each
A. Boil 1 g of the powdered tablets with 10 ml of 1 M sodium
of 30 ml, of chloroform, washing each extract with the same
hydroxide, cool and filter. Acidify the filtrate with 1 M sulphuric
10 ml of water. Filter the combined chloroform extracts, if
acid; a white precipitate is produced. Dissolve the precipitate
necessary, through absorbent cotton previously moistened
in about 2 ml of water and add ferric chloride test solution; a
with chloroform. Evaporate the solution to dryness and
deep violet colour is produced.
dissolve the residue as completely as possible in water,
B. Shake 0.5 g of the powdered tablets with 10 ml of water for warming gently if necessary. Cool, add sufficient water to
5 minutes, filter and add 10 ml of 1 M sodium hydroxide. produce 100.0 ml, mix and filter if necessary. Measure the
Extract with three quantities, each of 30 ml, of chloroform, absorbance of the resulting solution at the maximum at about
washing each extract with the same 10 ml of water. Filter the 273 nm (2.4.7).
combined extracts through absorbent cotton and evaporate
Calculate the content of C8H10N4O2 taking 504 as the specific
the filtrate to dryness. Reserve a quantity of the residue for
absorbance at 273 nm.
test C. Dissolve 10 mg of the residue in 1 ml of hydrochloric
acid, add 0.1 g of potassium chlorate and evaporate to Storage. Store protected from moisture.
dryness in a porcelain dish; a reddish residue remains which
becomes purple on exposure to ammonia vapour.
Atenolol
C. When examined in the range 230 nm to 360 nm (2.4.7), a
0.001per cent w/v solution of the residue reserved in Test B OH H
shows an absorption maximum at about 273 nm. O N CH3
O
Tests CH3
H2 N
Salicylic acid. Shake a quantity of the powdered tablets
containing 0.5 g of Aspirin with 50.0 ml of chloroform and 10 C14H22N2O3 Mol. Wt. 266.3
129
ATENOLOL IP 2007
Atenolol is (RS)-4-(2-hydroxy-3- Reference solution (b). Dissolve 50 mg of atenolol impurity

isopropylaminopropoxy)phenylacetamide. standard RS in 0.1 ml of dimethyl sulphoxide, if necessary
Atenolol contains not less than 99.0 per cent and not more heating the mixture by placing the container in a water-bath
than 101.0 per cent of C14H22N2O3, calculated on the dried for a few seconds and dilute to 25 ml with the mobile phase.
basis. Chromatographic system
Description. A white or almost white powder.
Identification ceramic microparticles (5 µm),
– mobile phase: dissolve 1.0 g of sodium octane-
Test A may be omitted if tests B and C are carried out. Tests B sulphonate and 0.4 gm of tetrabutyl ammonium
and C may be omitted if test A is carried out. hydrogen sulphate in 1000 ml of a mixture of 20 volumes
A. Determine by infrared absorption spectrophotometry (2.4.6). of tetrahydrofuran, 180 volumes of methanol and 800
Compare the spectrum with that obtained with atenolol RS or volumes of a 0.34 per cent w/v solution of potassium
with the reference spectrum of atenolol. dihydrogen phosphate and adjust the pH to 3.0 with
phosphoric acid,
0.01 per cent w/v solution in methanol shows absorption
maxima at about 275 nm and 282 nm. The ratio of the absorbance
at the maximum at about 275 nm to that at the maximum at
about 282 nm is 1.15 to 1.20. Equilibrate the column with the mobile phase for about
30 minutes and adjust the sensitivity of the system so that the
C. Determine by thin-layer chromatography (2.4.17), coating
height of the principal peak in the chromatogram obtained
the plate with silica gel GF254.
with reference solution (a) is at least 50 per cent of the full
Mobile phase. A mixture of 99 volumes of methanol and scale of the recorder.
1 volume of strong ammonia solution.
Inject reference solution (b). The resulting chromatogram is
Test solution. Dissolve 1.0 g of the substance under similar to that of the specimen chromatogram provided with
examination in sufficient methanol to produce 100 ml. atenolol impurity standard RS in that the peak due to bis-
Reference solution. A 1.0 per cent w/v solution of atenolol RS ether precedes and is separated from that due to tertiary amine,
in methanol. which normally appears as a doublet. If necessary, adjust the
concentration of sodium octanesulphonate; if its
Apply to the plate 10 µl of each solution. Allow the mobile concentration is increased, the retention time of the tertiary
phase to rise 10 cm. Dry the plate in a current of warm air and amine is prolonged.
examine in ultraviolet light at 254 nm. The principal spot in the
chromatogram obtained with the test solution corresponds to Inject separately test solution (a) and reference solution (a).
that in the chromatogram obtained with the reference solution. Continue the chromatography for four times the retention time
of the principal peak. The area of any secondary peak in the
Tests chromatogram obtained with test solution (a) is not greater
than half the area of the principal peak obtained with reference
Appearance of solution. A 1.0 per cent w/v solution is clear solution (a) (0.25 per cent); the sum of the areas of any
(2.4.1), and not more intensely coloured than degree 6 of the secondary peaks is not greater than the area of the principal
appropriate range of reference solutions. peak in the chromatogram obtained with reference solution
Related substances. Determine by liquid chromatography (a) (0.5 per cent). Ignore any peak with an area less than
(2.4.14). 10 per cent of that of the principal peak in the chromatogram
obtained with reference solution (a) (0.05 per cent).
Test solution (a). Dissolve 50 mg of the substance under
examination in 20 ml of the mobile phase and dilute to 25 ml If the substance under examination is found to contain more
with the mobile phase. than 0.15 per cent of bis-ether, its compliance is confirmed by
repeating the chromatography using 10 µl of test solution (b).
Test solution (b). Dissolve 50 mg of the substance under
examination in 0.1 ml of dimethyl sulphoxide, if necessary Chlorides (2.3.12). Dissolve 0.25 g in a mixture of 1 ml of 2 M
heating the mixture by placing the container in a water-bath nitric acid and 15 ml of water. The solution complies with the
for a few seconds and dilute to 25 ml with the mobile phase. limit test for chlorides without further addition of 2 M nitric
acid (0.1 per cent).
Reference solution (a). Dilute 0.5 ml of the test solution (a) to
100 ml with the mobile phase. Sulphated ash (2.3.18). Not more than 0.1 per cent.
130
IP 2007 ATORVASTATIN CALCIUM
Loss on drying (2.4.19). Not more than 0.5 per cent, determined – mobile phase: dissolve 0.8 g of sodium
on 1.0 g by drying in an oven at 105°. octanesulphonate and 0.4 gm of tetrabutylammonium
Assay. Weigh accurately about 0.2 g and dissolve in 80 ml of hydrogen sulphate in 1000 ml of a mixture of 20 volumes
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric of tetrahydrofuran, 180 volumes of methanol and 800
acid, determining the end-point potentiometrically (2.4.25). volumes of a 0.34 per cent w/v solution of potassium
Carry out a blank titration. dihydrogen phosphate and adjust the pH to 3.0 with
phosphoric acid,
1 ml of 0.1 M perchloric acid is equivalent to 0.02663 g of – flow rate. 1 ml per minute,
C14H22N2O3. – spectrophotometer set at 226 nm,
Inject each solution. The test is not valid unless the
Atenolol Tablets chromatogram obtained with reference solution (b) resembles
Atenolol Tablets contain not less than 92.5 per cent and not the reference chromatogram supplied with the atenolol impurity
more than 107.5 per cent of the stated amount of atenolol, standard RS in that the peak due to bis-ether precedes and is
C14H22N2O3. The tablets may be coated. separated from that due to tertiary amine, which is normally a
doublet. If necessary, adjust the concentration of sodium
Identification octanesulphonate in the mobile phase; if its concentration is
A. Heat a quantity of the powdered tablets containing about increased, the retention time of the tertiary amine is prolonged.
0.1 g of Atenolol with 15 ml of methanol to 50°, shake for In the chromatogram obtained with the test solution, the area
5 minutes, filter (Whatman No. 42 paper is suitable) and of any peak corresponding to 4-(2-hydroxy-3-isopropylamino-
evaporate the filtrate to dryness on a water-bath. Warm the propoxy)phenylacetic acid (blocker acid) is not greater than
residue with 10 ml of 0.1 M hydrochloric acid, shake and the area of the peak in the chromatogram obtained with
filter. Add to the filtrate sufficient 1 M sodium hydroxide to reference solution (a) (0.5 per cent) and the area of any peak
make it alkaline, extract with 10 ml of chloroform, dry by shaking corresponding to either tertiary amine or bis-ether is not greater
with anhydrous sodium sulphate, filter, evaporate the filtrate than half the area of the peak in the chromatogram obtained
to dryness on a water-bath and dry the residue at 105° for with reference solution (a) (0.25 per cent).
1hour. The residue complies with the following test. Other tests. Comply with the tests stated under Tablets.
Determine by infrared absorption spectrophotometry (2.4.6). Assay. Weigh and powder 20 tablets. Weigh accurately a
Compare the spectrum with that obtained with atenolol RS or quantity of the powder containing about 0.2 g of Atenolol,
with the reference spectrum of atenolol. transfer to a 500-ml volumetric flask using 300 ml of methanol,
B. When examined in the range 230 nm to 360 nm (2.4.7), the heat the resulting suspension to 60° and shake for 15 minutes.
solution obtained in the Assay shows absorption maxima at Cool, dilute to 500.0 ml with methanol, filter through a sintered-
about 275 nm and 282 nm. glass funnel (Porosil G3) and dilute a suitable volume of the
filtrate with sufficient methanol to produce a solution
Tests containing 0.01 per cent w/v of Atenolol. Measure the
Related substances. Determine by liquid chromatography absorbance of the resulting solution at the maximum at about
(2.4.14). 275 nm (2.4.7). Calculate the content of C14H22N2O3 taking 53.7
as the value of the specific absorbance at 275 nm.
containing 25 mg of Atenolol with 25 ml of the mobile phase
and mix with the aid of ultrasound for 20 minutes, filter (such
as Whatman GF/C filter) and use the filtrate. Atorvastatin Calcium
Reference solution (a). Dilute 1 volume of the test solution to
200 volumes with the mobile phase. H3C CH3
O OH OH
Reference solution (b). Dissolve 10 mg of atenolol impurity COO
standard RS in 0.1 ml of dimethyl sulphoxide with the aid of N N
Ca H , 3H2O
gentle heat, dilute to 10 ml with the mobile phase and mix.
– a stainless steel column 15 cm x 4.6 mm, packed with F
2
ceramic microparticles (5 µm), C66H68CaF2N4O10 Mol. Wt. 1155.36
131
ATORVASTATIN CALCIUM IP 2007
Atorvastatin Calcium is calcium salt of (βR,8R)-2-(4- Time Flow rate Mobile Mobile
fluorophenyl)-α,δ-dihydroxy-5-(1-methylethyl)-3-phenyl-4- phase B phase C
[(phenylamino)carbonyl]-1H-pyrrole-1-heptanoic acid (in min.) (ml per minute) (per cent v/v) (per cent v/v)
trihydrate. 0 1.8 100 0
Atorvastatin Calcium contains not less than 98.0 per cent and 20 1.8 100 0
not more than 102.0 per cent of C66H68 CaF2N4O10, calculated 35 1.5 25 75
on the anhydrous basis. 40 1.5 25 75
Description. A white to off-white, crystalline powder. 55 1.5 0 100
60 1.8 100 0
Identification
Determine by infrared absorption spectrophotometry (2.4.6). column efficiency is not less than 10000 theoretical plates and
Compare the spectrum with that obtained with atorvastatin the tailing factor is not more than 1.5.
calcium RS or with the reference spectrum of atorvastatin Inject alternatively the test solution and reference solution
calcium. (b). In the chromatogram obtained with the test solution, the
area of any individual secondary peak is not more than half
Tests the area of the peak in the chromatogram obtained with
Specific optical rotation (2.4.22). - 6.0º to -12.0º, determined in reference solution (b) (0.5 per cent) and the sum of the areas
a 1.0 per cent w/v solution in dimethylsulphoxide. of all the secondary peaks is not more than 2 times the area of
the peak in the chromatogram obtained with reference solution
Related substances. Determine by liquid chromatography (b) (2.0 per cent). Ignore any peak with an area less than 0.05
(2.4.14). times the area of the peak obtained in the chromatogram
Solvent mixture. A mixture of 40 volumes of acetonitrile and obtained in the chromatogram obtained with reference solution
60 volumes of water. (b) (0.05 per cent)
Test solution. Dissolve 50 mg of the substance under Heavy metals (2.3.13). 1.0 g complies with the limit test for
examination in 10 ml of methanol and dilute to 100 ml with the heavy metals, Method A (20 ppm).
solvent mixture. Water (2.3.43). 3.0 per cent to 7.0 per cent, determined on 0.3
g.
atorvastatin calcium RS in methanol. Dilute 5 ml of the Assay. Determine by liquid chromatography (2.4.14).
solution to 50 ml with the solvent mixture. Solvent mixture. A mixture of 40 volumes of acetonitrile and
Reference solution (b). Dilute 1 ml of reference solution (a) to 60 volumes of water.
100 ml with the solvent mixture. Test solution. Dissolve 80 mg of the substance under
Chromatographic system examination in 20 ml of methanol and dilute to 200 ml with the
– a stainless steel column 25 cm x 4.6 mm, packed with solvent mixture. Dilute this solution with the solvent mixture
octadecylsilane bonded to porous silica (5 µm) to produce a solution containing 0.008 per cent w/v of
– mobile phase: A. a mixture of 92.5 volumes of Atorvastatin Calcium.
acetonitrile and 7.5 volumes tetrahydrofuran, Reference solution. Dissolve 20 mg of atorvastatin calcium
B. a mixture of 58 volumes of a buffer RS in 5 ml of methanol and dilute to 50 ml with the solvent
solution prepared by dissolving 5.75 g of ammonium mixture. Dilute the solution with the solvent mixture to produce
dihydrogen orthophosphate in 1000 ml of water and 42 a solution containing 0.008 per cent w/v of Atorvastatin
volumes of mobile phase A, Calcium.
C. a mixture of 20 volumes of the buffer Chromatographic system
solution, 20 volumes of mobile phase A and 60 volumes – a stainless steel column 25 cm x 4.6 mm, packed with
of methanol, octadecylsilane bonded to porous silica (5 µm),
– a linear gradient programme using the conditions given – mobile phase: a mixture of 58 volumes of a buffer solution
below, prepared by dissolving 5.75 g of ammonium dihydrogen
– spectrophotometer set at 246 nm, orthophosphate in 1000 ml of water and 42 volumes of
– 20 µl loop injector. a mixture of 92.5 volumes of acetonitrile and 7.5 volumes
– injection delay 10 minutes, of tetrahydrofuran.,
132
IP 2007 ATORVASTATIN TABLETS
– flow rate. 1.8 ml per minute, Related substances. Determine by liquid chromatography
– spectrophotometer set at 246 nm, (2.4.14).
Solvent mixture. A mixture of 40 volumes of acetonitrile and
60 volumes of the buffer solution (see below).
relative standard deviation for replicate injections is not more
than 2.0 per cent, Test solution. Weigh accurately a quantity of the powdered
Inject alternatively the test solution and the reference solution. tablets containing 50 mg of atorvastatin, disperse in 10 ml of
methanol, add 20 ml of the solvent mixture, disperse with the
Calculate the content of C66H68 CaF2N4O10. aid of ultrasound, if required, and dilute to 100 ml with the
Storage. Store protected from light at a temperature not solvent mixture and filter.
exceeding 30º. Reference solution (a). Weigh accurately a suitable quantity
of atorvastatin calcium RS, dissolve in 5 ml of methanol and
dilute to 50 ml with the solvent mixture, to produce 0.05 percent
of atorvastatin
Atorvastatin Tablets Reference solution (b). Dilute 1 ml of reference solution (a)
Atorvastatin Calcium Tablets to 100 ml with the solvent mixture
Atorvastatin Tablets contains not less than 90.0 per cent and Chromatographic system
not more than 110.0 per cent of the stated amount of – a stainless steel column 25 cm x 4.6 mm, packed with
atorvastatin, C66H68 F2N4O10. octadecylsilane bonded to porous silica (5 µm),
– mobile phase: A. a mixture of 92.5 volumes of
Identification acetonitrile and 7.5 volumes tetrahydrofuran,
B. a mixture of 58 volumes of a buffer
In the Assay, the principal peak in the chromatogram obtained
solution prepared by dissolving 5.75 g of ammonium
dihydrogen orthophosphate in 1000 ml of water and 42
volumes of mobile phase A,
Tests C. a mixture of 20 volumes of the buffer
solution, 20 volumes of mobile phase A and 60 volumes
Dissolution (2.5.2). of methanol,
Apparatus No. 1 – a linear gradient programme using the conditions given
below,
Medium. 900 ml of phosphate buffer pH 6.8 – spectrophotometer set at 246 nm,
Speed and time. 75 rpm for 30 minutes. – 20 µl loop injector.
– injection delay 10 minutes.
Withdraw a suitable volume of the medium and filter. Determine
by liquid chromatography (2.4.14). Time Flow rate Mobile Mobile
phase B phase C
Test solution. Use the filtrate, diluted if necessary, with the
(in min.) (ml per minute) (per cent v/v) (per cent v/v)
dissolution medium.
0 1.8 100 0
Reference solution. Weigh a suitable quantity of atorvastatin 20 1.8 100 0
calcium RS and dissolve in sufficient methanol to produce a
solution containing 0.088 per cent of atorvastatin. Dilute 10.0 35 1.5 25 75
ml of the resulting solution to 100.0 ml with the medium. 40 1.5 25 75
Use the chromatographic system described under the Assay, 55 1.5 0 100
Inject the reference solution. The test is not valid unless the 60 1.8 100 0
column efficiency is not less than 7000 theoretical plates, the Inject reference solution (a). The test is not valid unless the
tailing factor is not more than 1.5 and the relative standard column efficiency is not less than 10000 theoretical plates and
deviation for replicate injections is not more than 2.0 per cent, the tailing factor is not more than 1.5,
Calculate the content of C66H68 F2N4O10. Inject alternatively the test solution and reference solution
D. Not less than 70 per cent of the stated amount of C66H68 (b). In the chromatogram obtained with the test solution, the
F2N4O10. area of any secondary peak is not more than the area of the
133
ATROPINE METHONITRATE IP 2007
peak in the chromatogram obtained with reference solution Atropine Methonitrate

(b) (1.0 per cent) and the sum of the areas of all the secondary
peaks is not more than 4 times the area of the peak in the Methylatropine Nitrate
chromatogram obtained with reference solution (b) (4.0 per
cent). Ignore any peak with an area less than 0.05 times the H3C
area of the peak obtained with reference solution (b) (0.05 per N CH3
cent).
Other tests. Comply with the tests stated under Tablets. H
OH
O NO3
Solvent mixture. A solution prepared by dissolving 6.8 g of
potassium dihydrogen orthophosphate and 0.9 g of sodium O
hydroxide in 1000 ml of water and adjusting the pH to 6.8 with
phosphoric acid or sodium hydroxide.
C18H26N2O6 Mol. Wt. 366.4
a quantity of the powdered tablets containing about 80 mg of Atropine Methonitrate is (RS)-(1R,3r,5S)-8-methyl-3-
atorvastatin and disperse in sufficient methanol to produce a tropoyloxytropanium nitrate.
solution containing 0.016 per cent w/v of atorvastatn. Disperse Atropine Methonitrate contains not less than 99.0 per cent
with the aid of ultrasound, if required, and filter. Dilute the and not more than 101.0 per cent of C18H26N2O6, calculated on
filtrate with sufficient of the solvent mixture to produce a the dried basis.
solution containing 0.008 per cent w/v of atorvastatin.
Description. Colourless crystals or a white, crystalline powder.
atorvastatin calcium RS and dissolve in sufficient methanol Identification
to produce a solution containing 0.08 per cent of atorvastatin.
To 5 ml of this solution, add 20 ml of methanol and dilute to 50 Test A may be omitted if tests B, C, and D are carried out.
ml with the solvent mixture to produce a solution containing Tests B and C may be omitted if tests A and D are carried out.
0.008 per cent w/v of atorvastatin. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with atropine
methonitrate RS.
B. To 0.05 ml of a 1 per cent w/v solution add 0.05 ml of a
0.1 per cent w/v solution of diphenylamine in nitrogen-free
– mobile phase: a mixture of 50 volumes of a buffer solution
sulphuric acid; an intense blue colour is produced.
prepared by dissolving 1.54 g of ammonium acetate in
1000 ml of water and adjusting the pH to 4.0 with glacial C. To 2.5 ml of a 10 per cent w/v solution add 2.5 ml of water
acetic acid, and 50 volumes of a mixture of 92.5 volumes and 2 ml of dilute sodium hydroxide solution; no precipitate
of acetonitrile and 7.5 volumes of tetrahydrofuran, is produced.
– flow rate. 2 ml per minute, D. Add about 1 mg to 4 drops of fuming nitric acid and
– spectrophotometer set at 246 nm, evaporate to dryness on a water-bath; a yellow residue is
– a 20 µl loop injector, obtained. To the cooled residue add 2 ml of acetone and
Inject the reference solution. The test is not valid unless the 4 drops of a 3 per cent w/v solution of potassium hydroxide in
column efficiency is not less than 7000 theoretical plates, the methanol; a violet colour is produced.
tailing factor is not more than 1.5 and the relative standard
deviation for replicate injections is not more than 1.0 per cent. Tests
Inject alternatively the test solution and the reference solution. Appearance of solution. A 5.0 per cent w/v solution in carbon
dioxide-free water is clear (2.4.1) and not more intensely
Calculate the content of C66H68F2N4O10, in the tablets. coloured than reference solution BS8 (2.4.1).
Storage. Store protected from moisture at a temperature not pH (2.4.24). 6.0 to 7.5, determined in a 5.0 per cent w/v solution.
exceeding 30º.
Specific optical rotation (2.4.22). –0.25° to +0.05°, determined
Labelling. The label states the strength in terms of the in a 10.0 per cent w/v solution, using a 2-dm tube (distinction
equivalent amount of atorvastatin. from hyoscyamine).
134
IP 2007 ATROPINE SULPHATE
Silver. To 10 ml of a 10.0 per cent w/v solution add 0.1 ml of Atropine Sulphate
sodium sulphide solution. The solution is not more intensely
coloured than reference solution BS8 (2.4.1).
Halides (2.3.12). 15 ml of a 5.0 per cent w/v solution in carbon
dioxide-free water complies with the limit test for chlorides, CH3
N
using 0.3 ml of chloride standard solution (25 ppm Cl ) for
preparing the standard.
H
Apomethylatropine. A 0.1 per cent w/v solution in 0.01 M , H2SO4, H2O
OH
hydrochloric acid shows absorption maxima at about 252 nm
and 257 nm (2.4.7). The ratio of the absorbance at about 257 O
nm to that at about 252 nm is not less than 1.17.
O
2
(2.4.17) coating the plate with silica gel G.
Mobile phase. A mixture of 60 volumes of ethyl acetate, 15 (C17H23NO3) 2,H2SO4, H2O Mol. Wt. 694.8
volumes of anhydrous formic acid, 15 volumes of water and
Atropine Sulphate is (RS)-(1R,3r,5S)-3-tropoyloxytropanium
sulphate monohydrate.
Atropine Sulphate contains not less than 99.0 per cent and
under examination in methanol (90 per cent).
not more than 101.0 per cent of atropine sulphate, (C17H23NO3)2,
Reference solution. Dilute 5 ml of the test solution to 100 ml H2SO4, calculated on the anhydrous basis.
with methanol (90 per cent), mix and dilute 10 ml of the
Description. Colourless crystals or a white, crystalline powder;
resulting solution to 100 ml with methanol (90 per cent).
odourless.
phase to rise 10 cm. Dry the plate at 105° until the odour of the Identification
solvent is not detectable. Allow it to cool to room temperature
Test A may be omitted if tests B and C are carried out. Test B
and spray with dilute potassium iodobismuthate solution
may be omitted if tests A and C are carried out.
until spots appear. Any secondary spot in the chromatogram
obtained with test solution is not more intense than the spot A. Determine by infrared absorption spectrophotometry (2.4.6).
in the chromatogram obtained with the reference solution. Compare the spectrum with that obtained with atropine
sulphate RS or with the reference spectrum of atropine
sulphate.
B. To a 2 per cent w/v solution add sodium hydroxide solution,
filter and transfer the precipitate with water. Dry the precipitate
Assay. Weigh accurately about 0.3 g and dissolve in 50 ml of at 60°. To 5 mg of the residue add 5 drops of fuming nitric acid
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric and evaporate to dryness on a water-bath. Cool the faintly
acid, determining the end-point potentiometrically (2.4.25). yellow coloured residue and add 2 ml of acetone and 4 drops
Carry out a blank titration. of a 3 per cent w/v solution of potassium hydroxide in
1 ml of 0.1 M perchloric acid is equivalent to 0.03664 g of methanol; a violet colour is produced.
C18H26N2O6. C. A 5 per cent w/v solution gives the reactions of sulphates
Storage. Store protected from light. (2.3.1).
Tests
Specific optical rotation (2.4.22). –0.50° to +0.05°, determined
in a 10.0 per cent w/v solution, using a 2-dm tube (distinction
from hyoscyamine).
Apoatropine. Absorbance of a 0.1 per cent w/v solution in
0.01 M hydrochloric acid at about 245 nm, not more than 0.4
(2.4.7).
135
ATROPINE INJECTION IP 2007
Foreign alkaloids and decomposition products. Determine by Test solution. Evaporate a volume of the injection containing
thin-layer chromatography (2.4.17), coating the plate with 5 mg of Atropine Sulphate to dryness on a water-bath, triturate
silica gel G. the residue with 1 ml of ethanol (95 per cent), allow to stand
Mobile phase. A mixture of 90 volumes of acetone, 7 volumes and use the supernatant liquid.
of water and 3 volumes of strong ammonia solution. Reference solution. A 0.5 per cent w/v solution of atropine
sulphate RS in ethanol (95 per cent).
under examination in methanol. Apply to the plate 5 µl of each solution. Allow the mobile
phase to rise 10 cm. Dry the plate at 105° for 20 minutes, allow
Reference solution (a). Dilute 1 ml of the test solution to
it to cool to room temperature and spray with potassium
100 ml with methanol.
iodobismuthate solution. The principal spot in the
Reference solution (b). Dilute 25 ml of reference solution (a) chromatogram obtained with the test solution corresponds to
to 50 ml with methanol. that in the chromatogram obtained with the reference solution.
Apply to the plate 10 µl of each solution. Allow the mobile B. In the Assay, the principal peak in the chromatogram
phase to rise 10 cm. Dry the plate at 105° for 15 minutes. Allow obtained with reference solution (a) corresponds to the peak
it to cool to room temperature and spray with dilute potassium in the chromatogram obtained with reference solution (b).
iodobismuthate solution. Any secondary spot in the
chromatogram obtained with the test solution is not more Tests
intense than the spot in the chromatogram obtained with pH (2.4.24). 3.0 to 5.5.
reference solution (a) and not more than one such spot is
more intense than the spot in the chromatogram obtained Other tests. Complies with the tests stated under Parenteral
with reference solution (b). Preparations (Injections).
Sulphated ash (2.3.18). Not more than 0.2 per cent. Assay. Determine by gas chromatography (2.4.13).
Water (2.3.43). Not more than 4.0 per cent, determined on Test solution. Add 1.0 ml of a 0.25 per cent w/v solution of
0.5 g. homatropine hydrobromide RS (internal standard) in methanol
(solution A), 1 ml of 5 M ammonia to a volume of the injection
Assay. Weigh accurately about 0.5 g, dissolve in 30 ml of containing 2.5 mg of Atropine Sulphate, diluted if necessary
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric to 5 ml with water and extract with two quantities, each of
acid, determining the end-point potentiometrically (2.4.25). 10 ml, of 0.1 M hydrochloric acid. Wash the combined extracts
Carry out a blank titration. with 5 ml of chloroform, shake the combined extracts with 1 g
1 ml of 0.1 M perchloric acid is equivalent to 0.06768 g of of anhydrous sodium sulphate, filter and evaporate the filtrate
(C17H23NO3)2, H2SO4. to dryness. Dissolve the residue in 5.0 ml of dichloromethane.
To 1.0 ml of this solution add 0.2 ml of a mixture of 4 volumes
of N,O-bis(trimethylsilyl)acetamide and 1 volume of
trimethylchlorosilane, mix and allow to stand for 30 minutes.
Reference solution (a). Prepare in the same manner as the test
Atropine Injection solution but omitting the addition of solution A.
Reference solution (b). Add 1 ml of solution A and 1 ml of 5 M
Atropine Sulphate Injection
ammonia to 5.0 ml of a 0.05 per cent w/v solution of atropine
Atropine Injection is a sterile solution of Atropine Sulphate in sulphate RS.
Water for Injections.
Atropine Injection contains not less than 90.0 per cent and – a glass column 1.5 m x 4 mm, packed with acid-washed,
not more than 110.0 per cent of the stated amount of atropine silanised diatomaceous support (80 to 100 mesh) coated
sulphate, (C17H23NO3)2,H2SO4,H2O. with 3 per cent w/w of phenyl methyl silicone fluid
(50 per cent phenyl) ( such as OV-17),
Identification – temperature:
column. 220°,
A. Determine by thin-layer chromatography (2.4.17), coating inlet port and detector. 260°,
the plate with silica gel G. – flow rate. 30 ml per minute of the carrier gas.
Mobile phase. A mixture of 50 volumes of chloroform, Calculate the content of (C17H23NO3)2,H2SO4,H2O in the
40 volumes of acetone and 10 volumes of diethylamine. injection.
136
IP 2007 ATROPINE TABLETS
Storage. Store protected from light. filter and evaporate the filtrate to dryness. Dissolve the residue
in 5.0 ml of dichloromethane. To 1.0 ml of this solution add
0.2 ml of a mixture of 4 volumes of N,O-bis (trimethylsilyl)
acetamide and 1 volume of trimethylchlorosilane, mix and
Atropine Eye Ointment allow to stand for 30 minutes.
Atropine Sulphate Eye Ointment Reference solution (a). Prepare in the same manner as the test
solution but omitting the addition of solution A.
Atropine Eye Ointment is a sterile preparation of Atropine
Reference solution (b). Add 2.0 ml of solution A and 2.0 ml of
Sulphate in an eye ointment base.
5 M ammonia to 20.0 ml of a 0.05 per cent w/v solution of
Atropine Eye Ointment contains not less than 92.5 per cent atropine sulphate RS and complete the procedure described
and not more than 107.5 per cent of the stated amount of under the test solution beginning at the words “Extract with
atropine sulphate, (C17H23NO3)2,H2SO4,H2O. two quantities, each of 10.0 ml, of chloroform,.....”.
Identification Chromatographic system
– a glass column 1.5 m x 4mm, packed with acid-washed,
Determine by thin- layer chromatography (2.4.17), coating the silanised diatomaceous support (80 to 100 mesh) coated
plate with silica gel G. with 3 per cent w/w of phenyl methyl silicone fluid
Mobile phase. A mixture of 50 volumes of chloroform, (50 per cent phenyl) (such as OV-17),
40 volumes of acetone and 10 volumes of diethylamine. – temperature:
column. 220°,
Test solution. Dissolve a quantity of the ointment containing inlet port and detector. 260°,
10 mg of Atropine Sulphate as completely as possible in 10 ml – flow rate. 30 ml per minute of the carrier gas.
of light petroleum (40° to 60°) and extract with two quantities,
Calculate the content of (C17H23NO3)2,H2SO4,H2O in the
each of 10 ml, of 0.05 M sulphuric acid, washing each acid
ointment.
solution with the same 5 ml of light petroleum (40° to 60°).
Mix the acid solutions, make alkaline with dilute ammonia Storage. Store at a temperature not exceeding 30°.
solution, and extract with two quantities, each of 15 ml, of
chloroform. Remove the chloroform and dissolve the residue
in 2 ml of ethanol (95 per cent). Atropine Tablets
Reference solution. A 0.5 per cent w/v solution of atropine Atropine Sulphate Tablets
sulphate RS in ethanol (95 per cent).
Atropine Tablets contain not less than 90.0 per cent and not
Apply to the plate 5 µl of each solution. Allow the mobile more than 110.0 per cent of the stated amount of atropine
phase to rise 10 cm. Dry the plate at 105° for 20 minutes, allow sulphate, (C17H23NO3)2,H2SO4,H2O.
it to cool to room temperature and spray with potassium
iodobismuthate solution. The principal spot in the Identification
that in the chromatogram obtained with the reference solution. A. Determine by thin- layer chromatography (2.4.17), coating
the plate with silica gel G.
Tests Mobile phase. A mixture of 50 volumes of chloroform,
Other tests. Complies with the tests stated under Eye 40 volumes of acetone and 10 volumes of diethylamine.
Ointments. Test solution. Shake a quantity of the powdered tablets
Assay. Determine by gas chromatography (2.4.13). containing 5 mg of Atropine Sulphate with 1 ml of ethanol
(95 per cent), centrifuge and use the supernatant liquid.
Test solution (a). Dissolve a quantity of the ointment
containing about 10 mg of Atropine Sulphate in 15 ml of Reference solution. A 0.5 per cent w/v solution of atropine
chloroform, add 2 ml of 0.5 per cent w/v solution of sulphate RS in ethanol (95 per cent).
homatropine hydrobromide RS (internal standard) in Apply to the plate 5 µl of each solution. Allow the mobile
methanol (solution A) and extract with two quantities, each of phase to rise 10 cm. Dry the plate at 105° for 20 minutes, allow
10.0 ml, of 0.1 M hydrochloric acid. Wash the combined it to cool to room temperature and spray with potassium
extracts with 10 ml of chloroform, add 2 ml of 5 M ammonia. iodobismuthate solution. The principal spot in the
Extract with two quantities, each of 10 ml, of chloroform, shake chromatogram obtained with the test solution corresponds to
the combined extracts with 2 g of anhydrous sodium sulphate, that in the chromatogram obtained with the reference solution.
137
AZATHIOPRINE IP 2007
B. The powdered tablets give the reactions of sulphates (2.3.1). test solution beginning at the words “shake the combined
extracts with 2 g of....”.
Tests
Reference solution (b). Prepare in the same manner as
Uniformity of content. Comply with the test stated under reference solution (a) but omitting the addition of solution A.
Tablets.
Determine by gas chromatography (2.4.13). – a glass column 1.5 m x 4 mm, packed with acid-washed,
Test solution. Powder one tablet and shake in a centrifuge silanised diatomaceous support (80 to 100 mesh) coated
tube with 5 ml of 0.1 M hydrochloric acid. Add 1 ml of a with 3 per cent w/w of phenyl methyl silicone fluid
0.06 per cent w/v solution of homatropine hydrobromide RS (50 per cent phenyl) ( such as OV-17),
(internal standard) in methanol (solution A), extract with two – temperature:
quantities, each of 5 ml, of chloroform and discard the column. 220°,
chloroform extracts. Add 1 ml of 5 M ammonia. Extract with inlet port and detector. 260°,
two quantities, each of 5 ml, of chloroform, shake the combined – flow rate. 30 ml per minute of the carrier gas.
extracts with 1 g of anhydrous sodium sulphate, filter and Calculate the content of (C17H23NO3)2,H2SO4,H2O in the tablets.
evaporate the filtrate to dryness. Dissolve the residue in
0.5 ml of a mixture of 20 volumes of dichloromethane
4 volumes of N, O-bis (trimethylsilyl)- acetamide and 1 volume
of trimethylchlorosilane, mix and allow to stand for 30 minutes. Azathioprine
Reference solution (a). Add 1 ml of solution A and 1 ml of 5 M NO2
ammonia to 5.0 ml of a 0.012 per cent w/v solution of atropine N
sulphate RS. Extract with two quantities, each of 5 ml, of
chloroform and complete the procedure described under N S
H
solution (1) beginning at the words “shake the combined H 3C N
N
extracts with 1 g of anhydrous sodium sulphate,.....”.
Reference solution (b). Prepare in the same manner as N N
reference solution (a) but omitting the addition of solution A.
C9H7N7O2S Mol. Wt. 277.3
Carry out the chromatographic procedure described under
Azathioprine is 6-[(1-methyl-4-nitro-1H-imidazol-5yl)
Assay.
sulphanyl]-7H-purine.
Calculate the content of (C17H23NO3)2,H2SO4,H2O in the tablet.
Azathioprine contains not less than 98.5 per cent and not
Other tests. Comply with the tests stated under Tablets. more than 101.0 per cent of C9H7N7O2S, calculated on the dried
Assay. Determine by gas chromatography (2.4.13). basis.
Test solution. Weigh and powder 20 tablets. Shake a quantity Description. A pale-yellow powder.
of the powdered tablets containing about 5 mg of Atropine
Sulphate with 10 ml of 0.1 M hydrochloric acid, add 1 ml of a Identification
0.5 per cent w/v solution of homatropine hydrobromide RS A. Determine by infrared absorption spectrophotometry (2.4.6).
(internal standard) in methanol (solution A), extract with two Compare the spectrum with that obtained with azathioprine
quantities, each of 10 ml, of chloroform and discard the RS.
chloroform extracts. Add 1 ml of 5 M ammonia. Extract with
two quantities, each of 10 ml, of chloroform, shake the B. Dissolve 0.15 g in 30 ml of dimethyl sulphoxide and dilute
combined extracts with 2 g of anhydrous sodium sulphate, to 500 ml with 0.1 M hydrochloric acid. Dilute 25 ml of this
filter and evaporate the filtrate to dryness. Dissolve the residue solution to 1000 ml with 0.1 M hydrochloric acid.
in 2.0 ml of dichloromethane. To 1.0 ml of this solution, add When examined in the range 230 nm to 350 nm (2.4.7), the
0.2 ml of a mixture of 4 volumes of N,O-bis(trimethylsilyl) resulting solution shows an absorption maximum at about 280
acetamide and 1 volume of trimethylchlorosilane, mix and nm; absorbance at 280 nm, about 0.600 to 0.660.
allow to stand for 30 minutes. C. To about 20 mg add 100 ml of water, heat and filter. To 5 ml
Reference solution (a). Add 1 ml of solution A and 1 ml of 5 M of the filtrate add 1 ml of hydrochloric acid and about 10 mg
ammonia to 10 ml of a 0.05 per cent w/v solution of atropine of zinc powder, stand for 5 minutes. The solution becomes
sulphate RS. Extract with two quantities, each of 10 ml, of yellow. Filter, cool in iced water, add 0.1 ml of sodium nitrite
chloroform and complete the procedure described under the solution and 0.1 g of sulphamic acid and shake until the
138
IP 2007 AZATHIOPRINE TABLETS
bubbles disappear. Add 1 ml of á-naphthol solution. A pale- Identification

pink precipitate is formed.
Tests the plate with cellulose F 254.
Acidity or alkalinity. To 0.5 g add 25 ml of carbon dioxide Mobile phase. A mixture of butan-1-ol saturated with 6 M
free water, shake for 15 minutes and filter. To 20 ml of the ammonia.
filtrate add 0.1 ml of methyl red solution. Not more than 0.2 ml Test solution. Shake a quantity of the powdered tablets
of 0.01 M hydrochloric acid or 0.01 M sodium hydroxide is containing 0.2 g of Azathioprine with 50 ml of 6 M ammonia,
required to change the colour of the indicator. filter through a glass micro fibre paper (such as Whatman GF/
Chloromethylnitroimidazole and mercaptopurine. Determine C) and use the filtrate.
by thin-layer chromatography (2.4.17), coating the plate with Reference solution. A 0.4 per cent w/v solution of
cellulose GF254. azathioprine RS in 6 M ammonia.
Mobile phase. Butanol saturated with dilute ammonia Apply to the plate 5 µl of each solution. After removal of the
solution. plate, dry the plate at 50° and examine in ultraviolet light at 254
NOTE – Prepare the following solutions immediately before nm. The principal spot in the chromatogram obtained with the
use. test solution corresponds to that in the chromatogram obtained
with the reference solution.
examination in?dilute ammonia solution and add sufficient B. Heat a quantity of the powdered tablets containing 20 mg
dilute ammonia solution to produce 10 ml. of Azathioprine with 100 ml of water and filter. To 5 ml of the
filtrate add 1 ml of hydrochloric acid and 10 mg of zinc powder
Reference solution (a). A 0.02 per cent w/v solution of and allow to stand for 5 minutes; a yellow colour is produced.
chloromethylnitroimidazole RS in dilute ammonia solution. Filter, cool in ice, add 0.1 ml of a 10 per cent w/v solution of
Reference solution (b). A 0.02 per cent w/v solution of sodium nitrite and 0.1 g of sulphamic acid and shake until the
mercaptopurine in dilute ammonia solution. bubbles disappear. Add 1 ml of 2-naphthol solution; a pale
pink precipitate is produced.
dry the plate at 50º and examine in ultraviolet light at 254 nm.
Tests
In the chromatogram obtained with the test solution, any spots
corresponding to chloromethylnitroimidazole and 5-Chloro-1-methyl-4-nitroimidazole and 6-mercaptopurine.
mercaptopurine are not more intense than the spots in the Determine by thin-layer chromatography (2.4.17), coating
chromatograms obtained with reference solution (a) (1.0 per the plate with cellulose F 254.
cent) and reference solution (b) (1.0 per cent).
Mobile phase. A mixture of butan-1-ol saturated with 6 M
Sulphated ash (2.3.18). Not more than 0.1 per cent. ammonia.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Test solution. Shake a quantity of the powdered tablets
on 0.5 g by drying in an oven at 105º. containing 0.2 g of Azathioprine with 10 ml of 6 M ammonia
Assay. Weigh accurately about 0.25 g and dissolve in 25 ml of and filter through a glass micro fibre filter paper (such as
dimethylformamide. Titrate with 0.1 M tetrabutylammonium Whatman GF/C).
hydroxide, determining the end-point potentiometrically Reference solution (a). A solution containing 2.0 per cent w/
(2.4.25). Carry out a blank titration. v of azathioprine RS and 0.02 per cent w/v of 6-mercapto-
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to purine in 6 M ammonia.
0.02773 g of C9H7N7O2S. Reference solution (b). A 0.02 per cent w/v solution of 6-
Storage. Store protected from light. mercaptopurine in 6 M ammonia.
Reference solution (c). A 0.02 per cent w/v solution of
chloromethylnitroimidazole RS in 6 M ammonia.
Azathioprine Tablets Apply to the plate 5 µl of each solution. After removal of the
plate, dry the plate at 50º and examine in ultraviolet light at 254
Azathioprine Tablets contain not less than 92.5 per cent and nm. Any spot in the chromatogram obtained with the test
not more than 107.5 per cent of the stated amount of solution corresponding to 6-mercaptopurine in the
azathioprine, C9H77O2S. chromatogram obtained with reference solution (a) is not more
139
AZITHROMYCIN IP 2007
intense than the spot in the chromatogram obtained with B. In the Assay, the principal peak in the chromatogram
reference solution (b). Any spot corresponding to 5-chloro-1- obtained with the test solution corresponds to the peak in the
methyl-4-nitroimidazole in the chromatogram obtained with chromatogram obtained with the reference solution.
the test solution is not more intense than the spot in the
chromatogram obtained with reference solution (c). Tests
Other tests. Comply with the tests stated under Tablets. Appearance of solution. Dissolve 0.5 g in anhydrous ethanol
Assay. Weigh and powder 20 tablets. Shake a quantity of the and dilute to 50.0 ml with the same solvent (solution A).
powder containing about 0.15 g of Azathioprine with 20 ml of Solution A is clear (2.4.1) and colourless (2.4.1).
dimethyl sulphoxide for 15 minutes and dilute to 500.0 ml with pH (2.4.24). 9.0 to 11.0 determined in a solution prepared by
0.1 M hydrochloric acid, filter. Dilute 25.0 ml of the filtrate to dissolving 0.1 g in 25.0 ml of methanol and further diluting to
1000.0 ml with 0.1 M hydrochloric acid. Measure the 50.0 ml with carbon dioxide- free water.
Specific optical rotation (2.4.22). – 45.0º to – 49.0º, determined
280 nm (2.4.7) using 0.1 M hydrochloric acid as the blank.
in solution A.
Calculate the content of C9H7N7O2S using a solution of
azathioprine RS of the same concentration in 0.1 M Related substances. Determine by liquid chromatography
hydrochloric acid. (2.4.14).
Storage. Store protected from light. Solvent mixture. 40 volumes acetonitrile and 60 volumes
water.
Azithromycin examination in the solvent mixture and dilute to 25.0 ml with
the solvent mixture.
Test solution (b). Dilute 5.0 ml of test solution (a) to 20.0 ml
CH3
with the solvent mixture.
H3C N
CH3
H3C OH
HO azithromycin RS in the solvent mixture.
CH3 HO
CH3 CH3
O O Reference solution (b). Dilute 1.0 ml of test solution (a) to
H3C CH3 O
O CH3 100.0 ml with the solvent mixture.
N OH O
CH3 O
OCH3 Reference solution (c). A solution containing 0.01 per cent w/
OH CH3
v of azithromycin RS and 6-demethyl-azithromycin RS
CH3 (azithromycin impurity A ) in the solvent mixture.
Reference solution (d). Dissolve the contents of a vial of 3-
C38H72N2O12 Mol. Wt. 749.0 deoxyazithromycin RS (azithromycin impurity B ) in 1.0 ml of
the solvent mixture. Use this solution for identification of the
Azithromycin is (2R,3S,4R,5R,8R,10R,11R,12S,13R,14R)-13-
peak due to impurity B.
[2,6-dideoxy-3-C-methyl-3-O-methyl-α-L-ribo-
hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy- Chromatographic system
3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3- – a stainless steel column 15 cm x 4.6 mm, packed with
(dimethylamino)-β-D-xylo- hexopyranosyl]oxy]-1-oxa-6- octadecylsilane chemically bonded to porous silica or
azacyclopentadecan-15-one. ceramic microparticles (5 µm),
– column temperature 70º,
Azithromycin contains not less than 94.0 per cent and not
– mobile phase: a mixture of 10 volumes of a 3.484 per cent
more than 102.0 per cent of C38H72N2O12, calculated on the
w/v solution of dipotassium hydrogen phosphate
anhydrous basis.
previously adjusted to pH 6.5 with phosphoric acid, 35
Description. A white or almost white powder. volumes of acetonitrile and 55 volumes of water,
Identification – spectrophotometer set at 215 nm,
A. Determine by infrared absorption spectrophotometry (2.4.6). – a 100 µl loop injector.
Compare the spectrum with that obtained with azithromycin The relative retention times with reference to azithromycin :
RS. impurity A, about 0.42; impurity B, about 1.7.
140
IP 2007 AZITHROMYCIN CAPSULES
Inject reference solution (c). The chromatogram obtained Test solution (a). Dissolve a suitable quantity of the mixed
shows peaks corresponding to azithromycin and azithromycin contents of 20 capsules containing about 0.1 g of Azithromycin
impurity A. The test is not valid unless the resolution between in the solvent mixture by shaking mechanically, dilute to 25.0
these peaks is at least 7.0. ml with the solvent mixture and filter.
Inject test solution (a) and reference solutions (b) and (d). Test solution (b). Dilute 5.0 ml of test solution (a) to 20.0 ml
Record the chromatogram of the test solution for 4.5 times the with the solvent mixture.
retention time of azithromycin. Reference solution (a). A 0.1 per cent w/v solution of
In the chromatogram obtained with test solution (a) the area azithromycin RS in the solvent mixture.
of any peak corresponding to impurity B is not more than Reference solution (b). Dilute 1.0 ml of test solution (a) to
twice the area of the principal peak in the chromatogram 100.0 ml with the solvent mixture.
obtained with reference solution (b) (2.0 per cent). The area of
Reference solution (c). A solution containing 0.01 per cent w/
any other secondary peak is not more than the area of the
principal peak in the chromatogram obtained with reference
(azithromycin impurity A) in the solvent mixture.
solution (b) (1.0 per cent). The sum of the areas of all the
secondary peaks is not more than 5 times the area of the Reference solution (d). Dissolve the contents of a vial of 3-
principal peak in the chromatogram obtained with reference deoxyazithromycin RS (azithromycin impurity B) in 1.0 ml of
solution (b) (5.0 per cent). Ignore any peak with an area 0.1 the solvent mixture. Use this solution for identification of the
times the area of the principal peak in the chromatogram peak due to impurity B.
obtained with reference solution (b) (0.1 per cent). Chromatographic system
Heavy metals (2.3.13). 0.8 g complies with the limit test for – a stainless steel column 15 cm x 4.6 mm, packed with
heavy metals, Method B (25 ppm). octadecylsilane chemically bonded to porous silica or
Sulphated ash (2.3.18). Not more than 0.2 per cent. – column temperature 70º,
Water (2,3,43). 1.8 per cent to 6.5 per cent, determined on 0.2 – mobile phase: a mixture of 10 volumes of a 3.484 per cent
g. w/v solution of dipotassium hydrogen phosphate with
the pH previously adjusted to 6.5 with phosphoric acid,
Assay. Determine by liquid chromatography (2.4.14) as
35 volumes of acetonitrile and 55 volumes of water,
described under Related Substances.
Inject alternately test solution (b) and reference solution (a). – spectrophotometer set at 215 nm,
Calculate the content of C38H72N2O12. – a 100 µl loop injector.
The relative retention times with reference to azithromycin:
Storage. Store protected from moisture.
impurity A, about 0.42; impurity B, about 1.7.
Inject reference solution (c). The chromatogram obtained
shows peaks corresponding to azithromycin and azithromycin
impurity A. The test is not valid unless the resolution between
Azithromycin Capsules these peaks is at least 7.0.
Azithromycin Capsules contain not less than 90.0 percent Inject test solution (a) and reference solutions (b) and (d).
and not more than 110.0 percent of the stated amount of Record the chromatogram of the test solution for 4.5 times the
azithromycin, (C38H72N2OI2). retention time of azithromycin.
Identification In the chromatogram obtained with test solution (a) the area
of any peak corresponding to impurity B is not more than 2
In the Assay, the principal peak in the chromatogram obtained times the area of the principal peak in the chromatogram
with the test solution corresponds to the peak in the obtained with reference solution (b) (2.0 per cent). The area of
chromatogram obtained with the reference solution. any other impurity peak is not more than the area of the
Tests solution (b) (1.0 per cent). The sum of the areas of all the
impurity peaks is not more than 5 times the area of the principal
Related substances. Determine by liquid chromatography peak in the chromatogram obtained with reference solution
(2.4.14). (b) (5.0 per cent). Ignore any peak with an area 0.1 times the
Solvent mixture. 40 volumes acetonitrile and 60 volumes area of the principal peak in the chromatogram obtained with
water. reference solution (b) (0.1 per cent).
141
AZITHROMYCIN ORAL SUSPENSION IP 2007
Dissolution (2.5.2). expected to be satisfactory for use, it contains not less than
Apparatus No. 1 80.0 per cent of the stated amount of azithromycin,
(C38H72N2OI2).
Medium. 900 ml of a buffer solution prepared by adding to 6
litres of 0.1 M dibasic sodium phosphate about 40 ml of The contents of the sealed container comply with the
hydrochloric acid to adjust the pH to 6.0, adding 600 mg of following test.
trypsin, and mixing. Water (2.3.43). Not more than 1.5 per cent, determined on
Speed and time. 100 rpm and 45 minutes. 0.5 g.
Withdraw a suitable volume of the medium and filter through
a filter having a porosity of 0.5 ìm or less. Storage. Store protected from moisture.
The constituted suspension complies with the tests stated
Determine by liquid chromatography (2.4.14) as described
under Oral liquids and with the following tests.
under Related substances using the following solutions.
Test solution. The filtrate from the dissolution vessel suitably Identification
diluted with the mobile phase if necessary.
In the Assay, the retention time of the principal peak in the
Reference solution. A solution of azithromycin RS in the chromatogram obtained with the test solution corresponds to
dissolution medium suitably diluted with the mobile phase to the peak in the chromatogram obtained with the reference
obtain a solution having the same concentration as that of the solution.
test solution.
Tests
Calculate the content of C38H72N2OI2 in the medium.
D. Not less than 75 per cent of the stated amount of pH (2.4.24). 8.5 to 11.0
C38H72N2OI2. Related substances. Determine by liquid chromatography
Water (2.3.43). Not more than 5.0 per cent determined on 0.2 g (2.4.14).
of the contents of the capsules. Solvent mixture. 40 volumes acetonitrile and 60 volumes
Other tests. Comply with the tests stated under Capsules. water.
Assay. Determine by liquid chromatography (2.4.14) as Test solution (a). Transfer an accurately weighed quantity of
described under Related Substances. the oral suspension containing about 0.1 g of Azithromycin to
a 25-ml volumetric flask, dissolve in the solvent mixture, dilute
Inject alternately test solution (b) and reference solution (a). to 25.0 ml with the solvent mixture and filter.
Calculate the content of C38H72N2O12 in the capsules. Test solution (b). Dilute 5.0 ml of test solution (a) to 20.0 ml
Storage. Store protected from moisture. Where packaged in with the solvent mixture.
unit-of-use containers, each container contains six 250-mg Reference solution (a). A 0.1 per cent w/v solution of
capsules and the label indicates the intended sequential day azithromycin RS in the solvent mixture.
of use for each capsule.
Reference solution (b). Dilute 1.0 ml of test solution (a) to
100.0 ml with the solvent mixture.
Azithromycin Oral Suspension v of azithromycin RS and 6-demethyl-azithromycin RS
Azithromycin Oral Suspension is a dry mixture of Azithromycin
with buffering agents and other excipients. It contains a Reference solution (d). Dissolve the contents of a vial of 3-
suitable flavouring agent. deoxyazithromycin RS (azithromycin impurity B) in 1.0 ml of
the solvent mixture. Use this solution for identification of the
peak due to impurity B.
before use. Chromatographic system
Azithromycin Oral Suspension contains not less than 90.0 per octadecylsilane chemically bonded to porous silica or
cent and not more than 110.0 per cent of the stated amount of ceramic microparticles (5 µm),
azithromycin, (C38H72N2OI2). – column temperature 70º,
When stored at the temperature and for the period stated on – mobile phase: a mixture of 10 volumes of a 3.484 per cent
the label during which the constituted suspension may be w/v solution of dipotassium hydrogen phosphate with
142
IP 2007 AZITHROMYCIN TABLETS
the pH previously adjusted to 6.5 with phosphoric acid, Test solution (a). Weigh and powder 20 tablets. Dissolve a
35 volumes of acetonitrile and 55 volumes of water, quantity of the powder containing about 0.1 g of Azithromycin
– flow rate. 1 ml per minute, in the solvent mixture by shaking mechanically, dilute to 25.0
– spectrophotometer set at 215 nm, ml with the solvent mixture and filter.
– a 100 µl loop injector. Test solution (b). Dilute 5.0 ml of test solution (a) to 20.0 ml
The relative retention times with reference to azithromycin: with the solvent mixture.
impurity A, about 0.42; impurity B, about 1.7. Reference solution (a). A 0.1 per cent w/v solution of
Inject reference solution (c). The chromatogram obtained azithromycin RS in the solvent mixture.
impurity A. The test is not valid unless the resolution between Reference solution (b). Dilute 1.0 ml of test solution (a) to
these peaks is at least 7.0. 100.0 ml with the solvent mixture.
Inject test solution (a) and reference solutions (b) and (d).
Record the chromatogram of the test solution for 4.5 times the
retention time of azithromycin.
Reference solution (d). Dissolve the contents of a vial of 3-
In the chromatogram obtained with test solution (a) the area deoxyazithromycin RS (azithromycin impurity B) in 1.0 ml of
of any peak corresponding to impurity B is not more than the solvent mixture. Use this solution for identification of the
twice the area of the principal peak in the chromatogram peak due to impurity B.
obtained with reference solution (b) (2.0 per cent). The area of
any other impurity peak is not more than the area of the Chromatographic system
principal peak in the chromatogram obtained with reference – a stainless steel column 15 cm x 4.6 mm, packed with
solution (b) (1.0 per cent). The sum of the areas of all the octadecylsilane chemically bonded to porous silica or
impurity peaks is not more than 5 times the area of the principal ceramic microparticles (5 µm),
peak in the chromatogram obtained with reference solution – column temperature 70º,
(b) (5.0 per cent). Ignore any peak with an area 0.1 times the – mobile phase: a mixture of 10 volumes of a 3.484 per cent
area of the principal peak in the chromatogram obtained with w/v solution of dipotassium hydrogen phosphate with
reference solution (b) (0.1 per cent). the pH previously adjusted to 6.5 with phosphoric acid,
35 volumes of acetonitrile and 55 volumes of water,
Assay. Determine by liquid chromatography (2.4.14) as – flow rate. 1 ml per minute,
described under Related Substances. – spectrophotometer set at 215 nm,
Inject alternately test solution (b) and reference solution (a). – a 100 µl loop injector.
Calculate the content of C38H72N2O12 in the suspension. The relative retention times with reference to azithromycin:
impurity A, about 0.42; impurity B, about 1.7.
Inject reference solution (c). The chromatogram obtained
Azithromycin Tablets impurity A. The test is not valid unless the resolution between
Azithromycin Tablets contain not less than 90.0 percent and these peaks is at least 7.0.
not more than 110.0 percent of the stated amount of Inject test solution (a) and reference solutions (b) and (d).
azithromycin, (C38H72N2OI2). Record the chromatogram of the test solution for 4.5 times the
retention time of azithromycin.
Identification
In the chromatogram obtained with test solution (a) the area
In the Assay, the retention time of the principal peak in the of any peak corresponding to impurity B is not more than
chromatogram obtained with the test solution corresponds to twice the area of the principal peak in the chromatogram
the peak in the chromatogram obtained with the reference obtained with reference solution (b) (2.0 per cent). The area of
solution. any other impurity peak is not more than the area of the
Tests solution (b) (1.0 per cent). The sum of the areas of all the
impurity peaks is not more than 5 times the area of the principal
peak in the chromatogram obtained with reference solution
(2.4.14).
(b) (5.0 per cent). Ignore any peak with an area 0.1 times the
Solvent mixture. 40 volumes acetonitrile and 60 volumes area of the principal peak in the chromatogram obtained with
water. reference solution (b) (0.1 per cent).
143
AZITHROMYCIN TABLETS IP 2007
Dissolution (2.5.2). obtain a solution having the same concentration as that of the
Apparatus No. 1 test solution.
Medium. 900 ml of a buffer solution prepared by adding to 6 Calculate the content of C38H72N2OI2 in the medium.
litres of 0.1 M dibasic sodium phosphate about 40 ml of D. Not less than 75 per cent of the stated amount of
hydrochloric acid to adjust the pH to 6.0, adding 600 mg of C38H72N2OI2.
trypsin, and mixing.
Speed and time. 100 rpm and 45 minutes. Water (2.3.43). Not more than 6.0 per cent determined on 0.2 g
of the powdered tablets.
a filter having a porosity of 0.5 ìm or less. Other tests. Comply with the tests stated under Tablets.
Determine by liquid chromatography (2.4.14) as described Assay. Determine by liquid chromatography (2.4.14) as
under Related substances using the following solutions. described under Related Substances.
Test solution. The filtrate from the dissolution vessel suitably Inject alternately test solution (b) and reference solution (a).
diluted with the mobile phase if necessary. Calculate the content of C38H72N2O12 in the tablets.
Reference solution. A solution of azithromycin RS in the
dissolution medium suitably diluted with the mobile phase to
144
B
Bacitracin ....
Bacitracin Zinc ....
Baclofen ....
Baclofen Oral Suspension ....
Baclofen Tablets ....
Barium Sulphate ....
Barium Sulphate For Suspension ....
Beclomethasone Dipropionate ....
Beclomethasone Inhalation ....
White Beeswax ....
Yellow Beeswax ....
Bentonite ....
Benzalkonium Chloride ....
Benzalkonium Chloride Solution ....
Benzathine Penicillin ....
Benzathine Penicillin Injection ....
Fortified Benzathine Penicillin Injection ....
Benzathine Penicillin Tablets ....
Benzhexol Hydrochloride ....
Benzhexol Tablets ....
Benzocaine ....
Benzoic Acid ....
Compound Benzoic Acid Ointment ....
Benzoin ....
Compound Benzoin Tincture ....
Benzyl Alcohol ....
Benzyl Benzoate ....
Benzyl Benzoate Application ....
Benzylpenicillin Potassium ....
Benzylpenicillin Sodium ....
145
Benzylpenicillin Injection ....

Betahistine Dihydrochloride ....
Betahistine Tablets ....
Betamethasone ....
Betamethasone Tablets ....
Betamethasone Sodium Phosphate ....
Betamethasone Eye Drops ....
Betamethasone Injection ....
Betamethasone Sodium Phosphate Tablets ....
Betamethasone Valerate ....
Betamethasone Valerate Ointment ....
Biperiden Hydrochloride ....
Biperiden Tablets ....
Bisacodyl ....
Bisacodyl Suppositories ....
Bisacodyl Tablets ....
Bismuth Subcarbonate ....
Bleomycin Sulpahte ....
Bleomycin Injection ....
Boric Acid ....
Bromhexine Hydrochloride ....
Bromhexine Tablets ....
Bromocriptine Mesylate ....
Bromocriptine Capsules ....
Bromocriptine Tablets ....
Bronopol ....
Budesonide ....
Bupivacaine Hydrochloride ....
Bupivacaine Injection ....
Buprenorphine Hydrochloride ....
Buprenorphine Injection ....
Buprenorphine Tablets ....
146
Busulphan ....
Busulphan Tablets ....
Butylated Hydroxyanisole ....
Butylated Hydroxytoluene ....
147
IP 2007 BACITRACIN ZINC
Bacitracin on 0.5 g by drying in an oven over phosphorus pentoxide at
Bacitracin is a mixture of polypeptides produced by the growth 60° at a pressure not exceeding 0.1 kPa for 3 hours.
of certain strains of Bacillus licheniformis and B. subtilis Assay. Determine by the microbiological assay of antibiotics,
(Fam. Bacillaceae). Its main components are Bacitracin A1, B1 Method A (2.2.10), and express the results in Units per mg.
and B2.
Bacitracin intended for administration as a spray in internal
Bacitracin has a potency of not less than 60 Units of bacitracin body cavities without a further appropriate procedure for
activity per mg, calculated on the dried basis. removal of bacterial endotoxins complies with the following
Description. A white or almost white powder; odourless or additional requirement.
with a faint odour; hygroscopic. Bacterial endotoxins (2.2.3). Not more than 0.01 Endotoxin
Identification Unit per unit.
Bacitracin intended for use in the manufacture of parenteral
A. Determine by thin-layer chromatography (2.4.17), coating preparations without a further appropriate sterilization
the plate with silica gel G. procedure complies with the following additional
Mobile phase. A mixture of 75 parts of phenol and 25 parts of requirement.
water. Sterility (2.2.11). Complies with the test for sterility.
Test solution. Dissolve 5 mg of the substance under Storage. Store protected from moisture at a temperature not
examination in a mixture of 0.5 ml of hydrochloric acid and exceeding 30°. If it is intended for use in the manufacture of
0.5 ml of water, heat in a sealed tube at 135° for 5 hours, ophthalmic preparations, the container should be sterile and
evaporate to dryness on a water-bath, continue to heat until sealed so as to exclude micro-organisms.
the odour of hydrogen chloride is no longer detectable and
dissolve the residue in 0.5 ml of water. Labelling. The label states (1) the number of Units per mg; (2)
whether or not the contents are intended for use in the
Reference solution. Prepare in the same manner as the test manufacture of ophthalmic preparations.
solution but using bacitracin zinc RS in place of the substance
under examination.
Apply to the plate 5 µl of each solution as bands 10 mm wide.
Place the plate in the tank so that it is not in contact with the Bacitracin Zinc
mobile phase and allow to stand for at least 12 hours before
Bacitracin Zinc is the zinc complex of Bacitracin.
development. Allow the mobile phase to rise 10 cm. Dry the
plate at 105°, spray with ethanolic ninhydrin solution and Bacitracin Zinc has a potency of not less than 60 Units of
heat at 110° for 5 minutes. The bands in the chromatogram bacitracin activity per mg, calculated on the dried basis.
obtained with the test solution correspond to those in the Description. A white or light yellowish-grey powder;
chromatogram obtained with the reference solution. odourless or with a faint odour; hygroscopic.
B. Shake 5 mg with 1 ml of water, add 1 ml of a 0.2 per cent
w/v solution of ninhydrin in 1-butanol and 0.5 ml of pyridine Identification
and heat at 100° for 5 minutes; a deep purple colour is
produced.
Tests Mobile phase. A mixture of 75 volumes of phenol and
25 volumes of water.
Appearance of solution. A 1.0 per cent w/v solution in carbon
dioxide-free water is clear, (2.4.1), and not more intensely Test solution. Dissolve 5 mg of the substance under
coloured than reference solution BYS6 (2.4.1). examination in a mixture of 0.5 ml of hydrochloric acid and
0.5 ml of water, heat in a sealed tube at 135° for 5 hours,
evaporate to dryness on a water-bath, continue to heat until
Bacitracin F and related substances. The ratio of the the odour of hydrogen chloride is no longer detectable and
absorbance (2.4.7) at the maximum at about 290 nm to that at dissolve the residue in 0.5 ml of water.
the maximum at about 252 nm of a 0.03 per cent w/v solution in
Reference solution. Prepare in the same manner as the test
0.05 M sulphuric acid is not more than 0.20.
solution but using bacitracin zinc RS in place of the substance
Sulphated ash (2.3.18). Not more than 3.0 per cent. under examination.
149
BACLOFEN IP 2007
Apply to the plate 5 µl of each solution as bands 10 mm wide. sterilization procedure complies with the following
Place the plate in the tank so that it is not in contact with the additional requirement.
mobile phase and allow to stand for at least 12 hours before Sterility (2.2.11). Complies with the test for sterility.
development. Allow the mobile phase to rise 10 cm. Dry the
plate at 105°, spray with ethanolic ninhydrin solution and Storage. Store protected from moisture at a temperature not
heat at 110° for 5 minutes. The bands in the chromatogram exceeding 30°. If it is intended for administration as a spray in
obtained with the test solution correspond to those in the internal body cavities, the container should be sterile and
chromatogram obtained with the reference solution. sealed so as to exclude micro-organisms.
B. Shake 5 mg with 1 ml of water, add 1 ml of a 0.2 per cent Labelling. The label states (1) the number of Units per mg; (2)
w/v solution of ninhydrin in 1-butanol and 0.5 ml of pyridine whether or not the contents are intended for administration as
and heat at 100° for 5 minutes; a deep purple colour is a spray in internal body cavities.
produced.
C. Ignite 0.15 g and allow to cool. The residue on dissolving in
1 ml of 2 M hydrochloric acid and diluting with 4 ml of water Baclofen
gives the reactions of zinc salts (2.3.1).
NH2
Tests
COOH
pH (2.4.24). 6.0 to 7.5, determined on the filtrate obtained by
shaking 1.0 g with 10 ml of carbon dioxide-free water.
Cl
Bacitracin F and related substances. The ratio of the
absorbance (2.4.7) at the maximum at about 290 nm to that at C10H12ClNO2 Mol. Wt. 213.7
the maximum at about 252 nm of a 0.03 per cent w/v solution in Baclofen is (3RS)-4-amino-3-(4-chlorophenyl)butanoic acid.
0.05 M sulphuric acid is not more than 0.15.
Baclofen contains not less than 98.0 per cent and not more
Zinc content. 4.0 per cent to 8.0 per cent, calculated on the than 101.0 per cent of C 10H12ClNO 2, calculated on the
dried basis, determined by the following method. Weigh anhydrous basis.
accurately about 0.2 g and dissolve in 20 ml of water and 3 ml
of strong ammonia-ammonium chloride solution and titrate Description. A white or almost white powder.
with 0.01 M disodium edetate using mordant black 11 mixture
as indicator.
Identification
1 ml of 0.01 M disodium edetate is equivalent to 0.000654 g of A. Determine by infrared absorption spectrophotometry (2.4.6).
Zn. Compare the spectrum with that obtained with baclofen RS.
Loss on drying (2.4.19). Not more than 5.0 per cent, determined B. When examined in the range 220 nm to 320 nm (2.4.7), a 0.07
on 0.5 g by drying in an oven over phosphorus pentoxide at per cent w/v solution shows three absorption maxima, at 259
60° at a pressure not exceeding 0.1 kPa for 3 hours. nm, 266 nm and 275 nm. The specific absorbances at these
maxima are 9.8 to 10.8, 11.5 to 12.7 and 8.4 to 9.3, respectively.
Assay. Weigh accurately about 0.1 g, suspend in 10 ml of
water and 0.5 ml of 2 M hydrochloric acid and add sufficient C. Determine by thin-layer chromatography (2.4.17), coating
water to produce 200.0 ml. Allow to stand at room temperature the plate with silica gel G..
for 30 minutes. Determine by the microbiological assay of Mobile phase. A mixture of 5 volumes of anhydrous formic
antibiotics, Method A (2.2.10), and express the results in Units acid, 5 volumes of water, 20 volumes of methanol, 30 volumes
per mg. of chloroform and 40 volumes of ethyl acetate.
Bacitracin Zinc intended for administration as a spray in Test solution. Dissolve 10 mg of the substance under
internal body cavities without a further appropriate examination in the mobile phase and dilute to 10 ml with the
procedure for removal of bacterial endotoxins complies with mobile phase.
the following additional requirement.
Reference solution. A 0.1 per cent w/v solution of baclofen
Bacterial endotoxins (2.2.3). Not more than 0.01 Endotoxin RS in the mobile phase.
Unit per unit. Apply to the plate 5 µl of each solution. Allow the mobile
Bacitracin Zinc intended for administration as a spray in phase to rise 12 cm. Dry the plate and spray with ninhydrin
internal body cavities without a further appropriate solution until the plate is slightly wet. Place the plate in an
150
IP 2007 BACLOFEN ORAL SOLUTION
oven maintained at 100º for 10 minutes. Examine in daylight. peak in the chromatogram obtained with reference solution
The principal spot in the chromatogram obtained with the test (c) (2.0 per cent).
solution corresponds to that in the chromatogram obtained Sulphated ash (2.3.18). Not more than 0.1 per cent.
Tests 1.0 g.
Appearance of solution. Dissolve 0.5 g in 1 M sodium
anhydrous acetic acid. Titrate with 0.1 M perchloric acid,
hydroxide and dilute to 25 ml with the same solvent. The
freshly prepared solution is not more intensely coloured than
reference solution BY5 (2.4.1).
C10H12ClNO2.
(2.4.14).
examination in the mobile phase and dilute to 10 ml with the
mobile phase.
Reference solution (a). A 0.25 per cent w/v solution of (4RS)-
Baclofen Oral Solution
4-(4-chlorophenyl)pyrrolidin-2-one RS (baclofen impurity Baclofen Oral Solution is a solution of Baclofen in a suitable
A) in the mobile phase. aqueous vehicle.
Reference solution (b). Dilute 1 ml of reference solution (a) to Baclofen Oral Solution contains not less than 95.0 per cent
100 ml with the mobile phase. and not more than 105.0 per cent of the stated amount of
baclofen, C10H12ClNO2.
Reference solution (c). Dilute 2 ml of the test solution to 100
ml with the mobile phase. Identification
Reference solution (d). Dilute 2 ml of the test solution and 2
ml of reference solution (a) to 100 ml with the mobile phase.
Chromatographic system Solvent mixture. 35 volumes of acetonitrile and 65 volumes
– a stainless steel column 25 cm x 4.0 mm, packed with of water.
octadecylsilyl silica gel (10 µm),
– mobile phase: a solution of 1.822 g of sodium Mobile phase. A mixture of 20 volumes of glacial acetic acid,
hexanesulphonate in 1000 ml of a mixture of 560 volumes 20 volumes of water and 80 volumes of butan-1-ol.
of water, 440 volumes of methanol and 5 volumes of Test solution. Dilute a volume of the oral solution containing
glacial acetic acid, 5 mg of Baclofen to 100 ml with the solvent mixture.
RS in the solvent mixture.
Adjust the sensitivity of the system so that the height of the
phase to rise 10 cm. Dry the plate in air. Place an evaporating
dish containing a mixture of 4 ml of water, 1 ml of 7 M
solution (c) is at least 50 per cent of the full scale of the recorder.
hydrochloric acid and 0.5 g of potassium permanganate in a
Inject reference solution (d). The test is not valid unless the
chromatography tank, close the tank and allow to stand for 2
resolution between the peaks corresponding to baclofen and
minutes. Place the plate in the tank, close the tank and leave
impurity A is at least 2.0.
the plate in contact with the vapour for 1 minute. After removal
Inject the test solution, reference solution (b) and reference of the plate, place it in a current of cold air until an area of
solution (c). Continue the chromatography for 5 times the coating below the line of application shows only a faint blue
retention time of the principal peak. In the chromatogram colour on the addition of 0.05 ml of potassium iodide and
obtained with the test solution, the area of the peak starch solution. Spray the plate with potassium iodide and
corresponding to baclofen impurity A is not greater than the starch solution and examine in daylight. The chromatogram
area of the principal peak in the chromatogram obtained with obtained with the test solution exhibits a spot that corresponds
reference solution (b) (1.0 per cent) and the sum of the areas to the spot in the chromatogram obtained with the reference
of all such peaks is not greater than the area of the principal solution.
151
BACLOFEN TABLETS IP 2007
B. In the Assay, the principal peak in the chromatogram Inject reference solution (b). The test is not valid unless the
obtained with the test solution corresponds to the peak in the resolution between the peaks due to baclofen impurity A and
chromatogram obtained with the reference solution. propyl-4-hydroxybenzoate is at least 5.0.
Tests Determine the weight per ml (2.4.29) of the oral solution and
Lactam. Determine by liquid chromatography (2.4.14) as calculate the content of C10H12ClNO2, weight in volume.
described under Assay using the following solutions. Storage. Store protected from light in a refrigerator (2º and 8º).
Test solution. Use the test solution prepared for the Assay. Do not freeze.
Reference solution (a). A solution containing 0.0002 per cent

w/v of (4RS)-4-(4-chlorophenyl)pyrrolidin-2-one RS
(baclofen impurity A) in the mobile phase. Baclofen Tablets
Reference solution (b). A solution containing 0.01 per cent w/
Baclofen tablets contain not less than 90.0 per cent and not
v of baclofen RS, 0.0003 per cent w/v of propyl 4-hydroxy-
more than 110.0 per cent of the stated amount of baclofen,
benzoate, 0.0003 per cent w/v of methyl 4-hydroxybenzoate
C10H12ClNO2.
and 0.0002 per cent w/v of baclofen impurity A in the mobile
phase.
Identification
resolution between the peaks due to methyl-4-hydroxybenzoate A. Determine by thin-layer chromatography (2.4.17), coating
and baclofen impurity A and between the peaks due to the plate with silica gel G.
baclofen impurity A and propyl-4-hydroxybenzoate is at least Solvent mixture. 4 volumes of absolute ethanol and 1 volume
5.0. of glacial acetic acid.
Inject the test solution and reference solution (b). In the Mobile phase. A mixture of 80 volumes of butan-1-ol, 20
chromatogram obtained with the test solution the area of any volumes of glacial acetic acid and 20 volumes of water.
peak corresponding to baclofen impurity A (lactam) is not
greater than the area of the peak in the chromatogram obtained
containing 20 mg of Baclofen with 20 ml of the solvent mixture
with reference solution (a) (2.0 per cent).
for 30 minutes and filter.
Other tests. Comply with the tests stated under Oral Liquids. Reference solution. A 0.1 per cent w/v solution of baclofen
Assay. Determine by liquid chromatography (2.4.14). RS in the solvent mixture.
Test solution. Dilute a weighed quantity of the oral solution Apply to the plate 5 µl of each solution. After development,
containing about 5 mg of Baclofen to 50.0 ml with the mobile dry the plate in air, spray with ninhydrin solution and heat at
phase. 100º for 10 minutes. The principal spot in the chromatogram
baclofen RS in the mobile phase.
Reference solution (b). A solution containing 0.01 per cent w/ obtained with the test solution corresponds to that in the
v of baclofen RS, 0.0003 per cent w/v of propyl 4-hydroxy- chromatogram obtained with the reference solution.
benzoate and 0.0002 per cent w/v of baclofen impurity A RS
in the mobile phase. Tests
Chromatographic system Lactam. Determine by liquid chromatography (2.4.14).
octadecylsilyl silica gel (10 µm) (such as Nucleosil C18), Test solution. Mix with the aid of ultrasound a quantity of the
– mobile phase: a solution prepared by dissolving 5 g of powdered tablets containing 0.1 g of Baclofen with 50 ml of
sodium dodecyl sulphate in a mixture of 5 ml of the mobile phase for 30 minutes, shaking occasionally to
orthophosphoric acid and 650 ml of water and diluting disperse the sample, and filter through a glass-fibre filter (such
to 1000 ml with acetonitrile, as Whatman GF/C).
– flow rate. 1.5 ml per minute, Reference solution (a). A solution containing 0.004 per cent
– spectrophotometer set at 218 nm, w/v of (4RS)-4-(4-chlorophenyl)pyrrolidin-2-one RS
– a 20 µl loop injector. (baclofen impurity A) in the mobile phase.
152
IP 2007 BARIUM SULPHATE
Reference solution (b). A solution containing 0.2 per cent w/ – mobile phase: 0.01 M sodium hexanesulphonate in a
v of baclofen RS and 0.004 per cent w/v of (4RS)-4-(4- mixture of 100 volumes of methanol, 100 volumes of
chlorophenyl)pyrrolidin-2-one RS (baclofen impurity A) in water and 1 volume of glacial acetic acid,
the mobile phase. – flow rate. 2 ml per minute,
Chromatographic system – spectrophotometer set at 265 nm,
– a stainless steel column 25 cm × 4.6 mm, packed with – a 20 µl loop injector.
octadecylsilyl silica gel (10 µm) (such as Spherisorb Inject alternately the test solution and the reference solution.
ODS 1), Calculate the content of C10H12ClNO2, in the tablets.
– mobile phase: a mixture of 5 volumes of glacial acetic
acid, 440 volumes of methanol and 560 volumes of
water containing 1.822 g per litre of sodium Barium Sulphate
hexanesulphonate,
BaSO4 Mol. Wt. 233.4
– spectrophotometer set at 266 nm, Barium Sulphate contains not less than 97.5 per cent and not
– a 20 µl loop injector. more than 100.5 per cent of BaSO4.
Inject reference solution (b). The test is not valid unless the Description. A fine, heavy, white powder, free from gritty
resolution between the peaks due to baclofen and baclofen particles; odourless.
impurity A is at least 2.0.
Inject the test solution and reference solution (a). In the Identification
chromatogram obtained with the test solution the area of any
A. Boil 0.2 g with 5 ml of a 50 per cent w/v solution of sodium
peak corresponding to baclofen impurity A (lactam) is not
carbonate for 5 minutes, add 10 ml of water and filter. Reserve
greater than the area of the peak in the chromatogram obtained
the residue for test B. Acidify the filtrate with dilute
with the reference solution (2.0 per cent).
hydrochloric acid; the solution gives the reactions of
Dissolution (2.5.2) sulphates (2.3.1).
Apparatus No 1 B. Wash the residue obtained in test A three times with
Medium. 900 ml of 0.1 M hydrochloric acid successive small quantities of water. To the residue add 5 ml
Speed and time. 50 rpm and 45 minutes. of dilute hydrochloric acid, filter and add to the filtrate 0.3 ml
of dilute sulphuric acid; a white precipitate is formed which
Withdraw a suitable volume of the medium and filter. is insoluble in dilute sodium hydroxide solution.
Determine by liquid chromatography (2.4.14) as described
under Assay using the following solutions. Tests
Test solution. Use the filtrate as given above. Acidity or alkalinity. Heat 5.0 g with 20 ml of carbon dioxide-
free water on a water-bath for 5 minutes and filter. To 10 ml of
the filtrate add 1 drop of bromothymol blue solution. Not
RS in the dissolution medium.
more than 0.5 ml of 0.01 M hydrochloric acid or 0.01 M sodium
Calculate the content of C10H12ClNO2 in the medium. hydroxide is required to change the colour of the solution.
Other tests. Comply with the tests stated under Tablets. Arsenic (2.3.10). Disperse 5.0 g in 50 ml of water and add 10 ml
Assay. Determine by liquid chromatography (2.4.14). of stannated hydrochloric acid AsT. The resulting solution
Test solution. Add a quantity of whole tablets containing 0.1
Heavy metals (2.3.13). Boil 4.0 g with a mixture of 2 ml of glacial
g of Baclofen to 25 ml of a mixture of 100 volumes of water and
acetic acid and 48 ml of water for 10 minutes. Add water to
1 volume of glacial acetic acid and disperse with the aid of
make upto 50 ml, filter and reject the first 5 ml of the filtrate.
ultrasound. Dilute to 50.0 ml with methanol, filter and use the
25 ml of the filtrate complies with the limit test for heavy metals,
filtrate.
Method A (10 ppm).
Phosphate. Boil 1 g with a mixture of 3 ml of nitric acid and
baclofen RS in a mixture of 100 volumes of methanol, 100
5 ml of water for 5 minutes and add water to restore the original
volumes of water and 1 volume of glacial acetic acid.
volume. Filter through a filter paper previously washed with
Chromatographic system dilute nitric acid. Add to the warm filtrate an equal volume of
– a stainless steel column 20 cm × 4.6 mm, packed with ammonium molybdate solution; no yellow precipitate is
octadecylsilyl silica gel (10 µm) (such as Nucleosil C18), formed.
153
BARIUM SULPHATE SUSPENSION IP 2007
Sulphide. Boil 10 g with a mixture of 10 ml of dilute of dilute hydrochloric acid, filter and add to the filtrate 0.3 ml
hydrochloric acid and 90 ml of water for 10 minutes. Expose of dilute sulphuric acid; a white precipitate is formed which
a lead acetate paper to the vapours; the paper does not is insoluble in dilute sodium hydroxide solution.
darken.
Tests
Acid-soluble substances. Cool the mixture obtained in the
test for Sulphide, add water to restore the original volume and pH (2.4.24). 4.0 to 8.0, determined in a 75.0 per cent w/v
filter through a filter paper previously washed with a mixture suspension in water.
of 10 ml of dilute hydrochloric acid and 90 ml of water,
returning the first portions, if necessary, to obtain a clear
on 1.0 g by drying in an oven at 105° for 4 hours.
filtrate. Evaporate 50 ml of the filtrate to dryness on a water-
bath and add 2 drops of hydrochloric acid and 10 ml of hot Assay. Weigh accurately about 0.6 g in a platinum crucible,
water. Filter again through acid-washed paper, prepared as add 5 g of sodium carbonate and 5 g of potassium carbonate
directed above, wash the filter paper with 10 ml of hot water and mix. Heat to 1000° and maintain at this temperature for
and evaporate the combined filtrate and washings. Dry the 15 minutes. Allow to cool and suspend the residue in 150 ml of
residue at 105°, cool and weigh (0.3 per cent). water. Wash the crucible with 2 ml of acetic acid and add to
the suspension. Cool in ice and filter by decantation,
Soluble barium salts. Digest the residue obtained in the test
transferring as little of the solid matter as possible to the filter.
for Acid-soluble substances with 10 ml of water and filter
Wash the residue with successive quantities of a 2 per cent
through a filter paper previously washed with a mixture of
w/v solution of sodium carbonate until the washings are free
10 ml of dilute hydrochloric acid and 90 ml of water. Add
from sulphate and discard the washings. Add 5 ml of dilute
0.5 ml of dilute sulphuric acid to the clear filtrate and set
hydrochloric acid to the filter and wash through into the
aside for 30 minutes; no turbidity is produced.
vessel containing the bulk of the solid matter with water. Add
Bulkiness. Place 5.0 g in a glass-stoppered 50-ml graduated 5 ml of hydrochloric acid and dilute to 100 ml with water. Add
cylinder having the 50-ml graduation mark 14 cm from the 10 ml of a 40 per cent w/v solution of ammonium acetate, 25 ml
base. Add water to 50 ml, shake the mixture for 5 minutes and of a 10 per cent w/v solution of potassium dichromate and
allow to stand for 15 minutes; it does not settle below the 10 g of urea. Cover, digest in an oven at 80° to 85° for 16 hours
15 ml mark. and filter while still hot through a sintered-glass filter (porosity
Loss on ignition. Not more than 2.5 per cent, determined on No. 4), washing the precipitate initially with a 0.5 per cent w/v
1.0 g at 600°. solution of potassium dichromate and finally with 2 ml of
water. Dry to constant weight at 105°.
1 g of the residue is equivalent to 0.9213 g of BaSO4.
Barium Sulphate Suspension
Barium Meal Beclomethasone Dipropionate
Barium Sulphate Suspension is a dry mixture of Barium
Sulphate with suitable flavours, colours, preservatives and O
suspending/dispersing agents. H3 C OCOC2H5
HO OCOC2H5
Barium Sulphate Suspension contains not less than 90.0 per H3C H CH3
barium sulphate, BaSO4. Cl H
Description. A white or coloured, fine powder or granules. O
Identification C28H37ClO7 Mol. Wt. 521.1

Beclomethasone Dipropionate is 9α-chloro-11β-hydroxy-16β-
A. Ignite 1 g to constant weight. Cool, boil 0.2 g of the residue
methyl-3,20-dioxopregna-1,4-diene-17,21-diyldipropionate.
with 5 ml of a 50 per cent w/v solution of sodium carbonate
for 5 minutes, add 10 ml of water and filter. Reserve the residue Beclomethasone Dipropionate contains not less than 96.0 per
for test B. Acidify the filtrate with dilute hydrochloric acid; cent and not more than 103.0 per cent of C28H37ClO7, calculated
the solution gives the reactions of sulphates (2.3.1). on the dried basis.
B. Wash the residue obtained in test A three times with Description. A white to creamy-white, crystalline powder;
successive small quantities of water. To the residue add 5 ml odourless.
154
IP 2007 BECLOMETHASONE INHALATION
Identification solution to 50.0 ml with the same solvent. Absorbance of the

resulting solution at the maximum at about 238 nm, 0.57 to 0.60
A. Determine by infrared absorption spectrophotometry (2.4.6). (2.4.7).
Compare the spectrum with that obtained with beclomethasone
dipropionate RS or with the reference spectrum of Sulphated ash (2.3.18). Not more than 0.1 per cent.
beclomethasone dipropionate. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
B. Determine by thin-layer chromatography (2.4.17), coating on 1.0 g by drying in an oven at 105° for 3 hours.
the plate with silica gel G. Assay. Determine by liquid chromatography (2.4.14).
Solvent mixture. A mixture of 90 volumes of acetone and Test solution. Weigh accurately about 70 mg of the substance
10 volumes of 1,2-propanediol. under examination, dissolve in methanol and dilute to 50.0 ml
Mobile phase. A mixture of 40 volumes of cyclohexane and 10 with same solvent. To 4.0 ml of this solution add 4.0 ml of a
volumes of toluene. 0.12 per cent w/v solution of testosterone propionate RS
(internal standard).
Reference solution. Dissolve an accurately weighed quantity
examination in 10 ml of the solvent mixture.
of beclomethasone dipropionate RS in methanol and dilute
Reference solution (a). Dissolve 25 mg of beclomethasone to obtain a solution having a known concentration of about
dipropionate RS in 10 ml of the solvent mixture. 1.4 mg per ml. To 4.0 ml of this solution add 4.0 ml of a 0.12
Reference solution (b). Mix equal volumes of the test solution per cent w/v solution of testosterone propionate RS (internal
and reference solution (a). standard).
Place the dry plate in a tank containing a shallow layer of the Chromatographic system
solvent mixture, allow the solvent mixture to ascend to the – a stainless steel column 25 cm x 4 mm, packed with
top, remove the plate from the tank and allow the solvent to octadecylsilyl silica gel (3 to 10 µm),
evaporate. Use within 2 hours, with the flow of the mobile – mobile phase: a mixture of 3 volumes of acetonitrile
phase in the direction in which the aforementioned treatment and 2 volumes of water, or such that the retention time
was done. of beclomethasone dipropionate is approximately 6
minutes and that of testsosterone propionate is
Apply to the plate 2 µl of each solution. Allow the mobile approximately 10 minutes,
phase to rise 12 cm. Dry the plate in a current of warm air, allow – flow rate. 1.5 ml per minute,
the solvent to evaporate, heat at 120° for 15 minutes and spray – spectrophotometer set at 254 nm,
the hot plate with ethanolic sulphuric acid (20 per cent v/v). – a 20 µl loop injector.
Heat at 120° for a further 10 minutes, allow to cool and examine
in daylight and in ultraviolet light at 365 nm. The principal Inject the reference solution. The test is not valid unless the
spot in the chromatogram obtained with the test solution relative standard deviation for replicate injections is not more
corresponds to that in the chromatogram obtained with than 3.0 per cent.
reference solution (a). The principal spot in the chromatogram Inject alternately the test solution and the reference solution.
obtained with reference solution (b) appears as a single,
Calculate the percentage content of C28H37ClO7.
compact spot.
C. Determine by the oxygen flask method (2.3.34), on 25 mg
and use a mixture of 20 ml of water and 1 ml of 1 M sodium
hydroxide as the absorbing liquid. The liquid gives reaction A
of chlorides (2.3.1). Beclomethasone Inhalation
D. In the Assay, the principal peak in the chromatogram Beclomethasone Dipropionate Inhalation;
obtained with the test solution corresponds to that in the Beclomethasone Inhalation Aerosol
Beclomethasone Inhalation is a suspension of Beclomethasone
Tests Dipropionate in a suitable liquid in a suitable pressurised
container.
Beclomethasone Inhalation delivers not less than 80.0 per
in a 1.0 per cent w/v solution in dioxan.
cent and not more than 120.0 per cent of the stated amount per
Light absorption. Dissolve 50.0 mg in sufficient ethanol inhalation of beclomethasone dipropionate, C28H37ClO7, by
(95 per cent) to produce 100.0 ml and dilute 2.0 ml of this actuation of the valve.
155
BECLOMETHASONE INHALATION IP 2007
Identification Assay. Carry out the test for Content of active ingredient
delivered per actuation stated under Inhalation Preparations
A. Discharge the container a sufficient number of times at low (Pressurised metered-dose Preparations).
relative humidity into a mortar to obtain about 2 mg of
anhydrous Beclomethasone Dipropionate. Heat at 110º for 2 Use 40 ml of dehydrated methanol as the solvent. Discharge
hours at a pressure of 2kPa, cool, grind the residue thoroughly the number of deliveries that constitute the minimum
with 0.1 g of potassium bromide, add a further 0.2 g of potassium recommended dose, keep the solution on a water-bath for 5
bromide and mix thoroughly. minutes to expel the propellants. Transfer the solution and
washings to a flask containing sufficient testosterone
On the resultant dispersion determine by infrared absorption
propionate RS (internal standard) in methanol that, on dilution
spectrophotometry (2.4.6). Compare the spectrum with that
to a suitable volume with appropriate amounts of water and
obtained with beclomethasone dipropionate RS or with the
methanol, the final solution contains 0.00015 per cent w/v
reference spectrum of beclometasone dipropionate.
each of testosterone propionate and beclomethasone
B. In the Assay, the principal peak in the chromatogram dipropionate in the methanol-water mixture in the proportions
obtained with the test solution corresponds to the peak due 70:30 by volume.
to beclometasone dipropionate in the reference solution.
Determine by liquid chromatography (2.4.14).
Tests Test solution. The diluted solution obtained as given above.
Related substances. Determine by thin-layer chromatography Reference solution. A solution containing 0.00015 per cent
(2.4.17), coating the plate with silica gel G. w/v each of the internal standard and beclomethasone
dipropionate RS in the mobile phase.
Mobile phase. A mixture of 3 volumes of methanol and 97
volumes of dichloroethane. Chromatographic system
Test solution. Discharge from the container into a small, dry – a stainless steel column 10 cm x 4.6 mm, packed with
flask a sufficient number of times to obtain 0.5 g of octadecylsilane bonded to porous silica (5 µm),
Beclometasone Dipropionate and dissolve the residue in 2 ml – column temperature. 50º
of acetone. Evaporate the solution to a volume such that the – mobile phase: a mixture of 70 volumes of methanol and
whole solution can be applied to the plate. 30 volumes of water, adjusted if necessary so that the
resolution between the peaks due to beclomethasone
Reference solution (a). A 0.1 per cent w/v solution of dipropionate and the internal standard is not less than
beclomethasone dipropionate RS in acetone. 2.0,
Reference solution (b). Dilute 5 ml of reference solution (a) to – flow rate. 2 ml per minute,
10 ml with acetone. – spectrophotometer set at 239 nm,
Reference solution (c). Dilute 5 ml of reference solution (a) to – a 20 µl loop injector.
20 ml with acetone. Inject the test solution and the reference solution. The test is
Apply to the plate 10 µl of each solution. After development, not valid unless the resolution between the two principal
dry the plate in air, spray with alkaline tetrazolium blue peaks in the chromatogram obtained with the reference
solution and heat at 50º for 5 minutes. Cool and spray again solution is at least 2.0.
with alkaline tetrazolium blue solution. Any secondary spot Calculate the amount of C28H37ClO7 delivered per actuation of
in the chromatogram obtained with the test solution is not the valve.
more intense than the spot in the chromatogram obtained
with reference solution (a), not more than one such spot is Determine the content of active ingredient a second and third
more intense than the spot in the chromatogram obtained time by repeating the procedure on the middle ten and on the
with reference solution (b) (1 per cent) and any other secondary last ten successive combined actuations of the valve. For
spot is not more intense than the spot in the chromatogram each of the three determinations the average content of
obtained with reference solution (c) (0.5 per cent). Ignore any C28H37ClO7 delivered per actuation of the valve meets the
spot with an Rf value of more than 0.85. requirements.
Other tests. Complies with the tests stated under Inhalation Storage. Store protected from moisture at a temperature not
Preparations (Pressurised metered-dose Preparations). exceeding 30º.
Follow the procedure described under Assay wherever the Labelling. The label states the amount of active ingredient
amount of active substance is to be determined in any test. delivered per inhalation.
156
IP 2007 YELLOW BEESWAX
White Beeswax Glycerin and other polyhydric alcohols. To 0.2 g add 10 ml of

ethanolic potassium hydroxide solution, heat under a reflux
White Beeswax is obtained by bleaching Yellow Beeswax. condenser in a water-bath for 30 minutes, add 50 ml of 1 M
Description. Yellowish-white pieces or plates, translucent when sulphuric acid, cool and filter. Rinse the flask and filter with
thin, with a fine-grained, matt, non-crystalline fracture; 1 M sulphuric acid, combine the filtrate and washings and
becomes soft and pliable when warmed by hand. Odour, faint dilute to 100 ml with 1 M sulphuric acid (solution A). Into two
and characteristic and similar to that of yellow beeswax. matched test-tubes introduce, respectively, 1 ml of solution A
and 1 ml of a 0.001 per cent w/v solution of glycerin in 1 M
Tests sulphuric acid (solution B). Add 0.5 ml of a 1.07 per cent w/v
solution of sodium periodate to each tube, mix, allow to stand
Melting range (2.4.21). 61° to 65°, determined by Method IV.
for 5 minutes, add to each tube 1 ml of decolorised fuchsin
Acid value (2.3.23). 5 to 15, determined by the following method. solution and mix; any precipitate disappears. Place the tubes
Weigh accurately about 5.0 g in a 250-ml conical flask fitted in a beaker containing water at 40° and observe for 10 to 15
with a reflux condenser, add 40 ml of xylene and a few glass minutes during cooling. Any bluish violet colour in the tube
beads, heat until dissolved, add 20 ml of ethanol (95 per containing solution A is not more intense than that in the tube
cent) and 0.5 ml of phenolphthalein solution and titrate the containing solution B (0.5 per cent w/w, calculated as glycerin).
hot solution with 0.5 M ethanolic potassium hydroxide until
a red colour persists for at least 10 seconds (n1 ml). Repeat the
procedure omitting the substance under examination (n2 ml). Yellow Beeswax
Calculate the Acid value from the expression 28.05(n1 - n2)/w,
where w is the weight, in g, of the substance taken. Yellow beeswax is the wax obtained by melting the walls of the
honeycomb of the bee, Apis mellifera Linn. with hot water
Ester value (2.3.26). 75 to 95, determined by subtracting the and removing the foreign matter.
Acid value from the Saponification value.
Description. Yellow or light brown pieces or plates, with a
Ratio number. The Ester value divided by the Acid value is fine-grained, matt, non-crystalline fracture; becomes soft and
between 5 and 19. pliable when warmed by hand. Odour, faint and characteristic.
Saponification value (2.3.37). 87 to 104, determined by the It is tasteless and does not stick to the teeth.
following method. Weigh accurately about 2.0 g, add 30 ml of
a mixture of equal volumes of xylene and ethanol (95 per Tests
cent) and a few glass beads, heat until dissolved, add 25.0 ml Melting range (2.4.21). 61° to 65°, determined by Method IV.
of 0.5 M ethanolic potassium hydroxide and heat under a
reflux condenser for 3 hours. Titrate the hot solution Acid value (2.3.23). 5 to 15, determined by the following method.
immediately with 0.5 M hydrochloric acid using 1 ml of Weigh accurately about 5.0 g in a 250-ml conical flask fitted
phenolphthalein solution as indicator, bringing the solution with a reflux condenser, add 40 ml of xylene and a few glass
back to boil several times during the titration (n1 ml). Repeat beads, heat until dissolved, add 20 ml of ethanol (95 per
the procedure omitting the substance under examination cent) and 0.5 ml of phenolphthalein solution and titrate the
(n2 ml). Calculate the Saponification value from the expression hot solution with 0.5 M ethanolic potassium hydroxide until
28.05(n2 - n1)/w, where w is the weight, in g, of the substance a red colour persists for at least 10 seconds (n1 ml). Repeat the
taken. procedure omitting the substance under examination (n2 ml).
Calculate the Acid value from the expression 28.05(n2 - n1)/w,
Fats, fatty acids, Japan wax and resin. Boil 5.0 g for 10 minutes where w is the weight, in g, of the substance taken.
with 80 ml of a 10 per cent w/v solution of sodium hydroxide,
replace the water lost by evaporation, cool, filter the solution Ester value (2.3.26). 75 to 95, determined by subtracting the
through a plug of glass wool and acidify with hydrochloric Acid value from the Saponification value.
acid; no precipitate is produced. Ratio number. The Ester value divided by the Acid value is
Ceresin, paraffin and other waxes. To 3.0 g in a 100-ml round- between 5 and 19.
bottomed flask add 30 ml of a 4 per cent w/v solution of Saponification value (2.3.37). 87 to 104, determined by the
potassium hydroxide in aldehyde-free ethanol (95 per cent) following method. Weigh accurately about 2.0 g, add 30 ml of
and boil gently under a reflux condenser for 2 hours. Remove a mixture of equal volumes of xylene and ethanol (95 per
the condenser and immediately insert a thermometer, place cent) and a few glass beads, heat until dissolved, add 25.0 ml
the flask in a water-bath at 80° and allow to cool with of 0.5 M ethanolic potassium hydroxide and heat under a
continuous swirling. The solution may be opalescent, but no reflux condenser for 3 hours. Titrate the hot solution
precipitate is formed before the temperature reaches 65°. immediately with 0.5 M hydrochloric acid using 1 ml of
157
BENTONITE IP 2007
phenolphthalein solution as indicator, bringing the solution gives the reactions of aluminium salts, (2.3.1). Add to the
back to boil several times during the titration (n1 ml). Repeat reserved filtrate and washings 3 ml of hydrochloric acid; a
the procedure omitting the substance under examination gelatinous precipitate is produced.
(n2 ml). Calculate the Saponification value from the expression
28.05(n2 - n1)/w, where w is the weight, in g, of the substance Tests
taken. pH (2.4.24). 9.0 to 10.5, determined in a 2.0 per cent w/v
Fats, fatty acids, Japan wax and resin. Boil 5 g for 10 minutes suspension in water.
with 80 ml of a 10 per cent w/v solution of sodium hydroxide, Heavy metals (2.3.13). To 5.0 g add 7.5 ml of 2 M hydrochloric
replace the water lost by evaporation, cool, filter the solution acid and 27.5 ml of water, boil for 5 minutes, centrifuge and
through a plug of glass wool and acidify with hydrochloric filter the supernatant liquid. Wash the residue with water,
acid; no precipitate is produced. filter, combine the filtrates and dilute to 50 ml with water. To
Ceresin, paraffin and other waxes. To 3.0 g in a 100-ml round- 5 ml of the solution add 5 ml of water, 10 ml of hydrochloric
bottomed flask add 30 ml of a 4 per cent w/v solution of acid and 25 ml of 4-methyl-2-pentanone, shake for 2 minutes,
potassium hydroxide in aldehyde-free ethanol (95 per cent) allow the layers to separate and evaporate the aqueous layer
and boil gently under a reflux condenser for 2 hours. Remove to dryness on a water-bath. Dissolve the residue in 1 ml of 5 M
the condenser and immediately insert a thermometer, place acetic acid, dilute to 25 ml and filter. The resulting solution
the flask in a water-bath at 80° and allow to cool with complies with the limit test for heavy metals, Method D
continuous swirling. The solution may be opalescent, but no (50 ppm). Prepare the standard using lead standard solution
precipitate is formed before the temperature reaches 65°. (1 ppm Pb).
Glycerin and other polyhydric alcohols. To 0.2 g add 10 ml of Sedimentation volume. In a mortar, mix 6.0 g with 0.3 g of light
ethanolic potassium hydroxide solution, heat under a reflux magnesium oxide, freshly calcined. Mix the powder
condenser in a water-bath for 30 minutes, add 50 ml of 1 M progressively with 200 ml of water. Shake for 1 hour and place
sulphuric acid, cool and filter. Rinse the flask and filter with 100 ml of the suspension in a 100-ml graduated cylinder. After
1 M sulphuric acid, combine the filtrate and washings and 24 hours the volume of the clear supernatant liquid is not
dilute to 100 ml with 1 M sulphuric acid (solution A). Into two greater than 2 ml.
matched test-tubes introduce, respectively, 1 ml of solution A Swelling power. Add 2.0 g in twenty portions at intervals of
and 1 ml of a 0.001 per cent w/v solution of glycerin in 1 M 2 minutes to 100 ml of a 1 per cent w/v solution of sodium
sulphuric acid (solution B). Add 0.5 ml of a 1.07 per cent w/v lauryl sulphate in a 100-ml graduated cylinder about 3 cm in
solution of sodium periodate to each tube, mix, allow to stand diameter. Allow each portion to settle before adding the next
for 5 minutes, add to each tube 1 ml of decolorised fuchsin and let it stand for 2 hours. The apparent volume of the
solution and mix; any precipitate disappears. Place the tubes sediment at the bottom of the cylinder is not less than 24 ml.
in a beaker containing water at 40° and observe for 10 to 15 Coarse particles. To 20 g add 1000 ml of water and mix for
minutes during cooling. Any bluish violet colour in the tube 15 minutes at not less than 5000 rpm. Transfer to a wet sieve of
containing solution A is not more intense than that in the tube nominal aperture of 75 mm, previously dried at 100° to 105°
containing solution B (0.5 per cent w/w, calculated as glycerin). and weighed, and wash with three quantities, each of 500 ml,
Storage. Store in well-closed containers. of water, ensuring that any agglomerates are dispersed. Dry
at 100° to 105° and weigh. The weight of the matter on the
sieve is not more than 0.1 g (0.5 per cent).
Bentonite Microbial contamination (2.2.9). 1g is free from Escherichia
coli.
Bentonite is a natural, colloidal, hydrated aluminium silicate
that has been processed to remove grit and non-swelling Loss on drying (2.4.19). Not more than 15.0 per cent,
components of the ore. determined on 1.0 g by drying in an oven at 105°.
Description. A very fine, pale buff or cream-coloured to
greyish-white powder, free or almost free from gritty particles. Benzalkonium Chloride
Identification Benzalkonium Chloride is a mixture of alkylbenzyl-
Fuse 1 g with 2 g of anhydrous sodium carbonate, warm the dimethylammonium chlorides, the alkyl groups having chain
residue with 10 ml of water, filter, wash the filter with 5 ml of lengths of C8 to C18.
water and reserve the combined filtrate and washings. Dissolve Benzalkonium Chloride contains not less than 95.0 per cent
the residue in 10 ml of dilute hydrochloric acid; the solution and not more than 104.0 per cent of alkylbenzyldimethyl-
158
IP 2007 BENZALKONIUM CHLORIDE SOLUTION
ammonium chlorides, calculated as C22H40ClN on the dried hydrochloric acid with 0.05 M potassium iodate in a similar
basis. manner; the difference between the titrations represents the
Description. A white or yellowish-white powder or gelatinous, amount of 0.05 M potassium iodate required.
yellowish-white fragments, hygroscopic, soapy to the touch. 1 ml of 0.05 M potassium iodate is equivalent to 0.0354 g of
C22H40ClN.
Identification
Storage. Avoid contact with metals.
A. Dilute 0.1 g with 10 ml of water. To 5 ml add 1.5 ml of dilute
nitric acid; a white precipitate is produced which is soluble in
ethanol (95 per cent). To the remainder add 1.5 ml of mercuric Benzalkonium Chloride Solution
chloride solution; a white precipitate is produced which is
soluble in ethanol (95 per cent). Benzalkonium Chloride Solution is a solution of a mixture of
alkylbenzyldimethylammonium chlorides, the alkyl groups
B. Dissolve 0.25 g in 1 ml of sulphuric acid, add 0.1 g of having chain lengths of C8 to C18. It may contain ethanol
potassium nitrate, heat on a water-bath for 5 minutes, cool, (95 per cent). In making Benzalkonium Chloride Solution, the
dilute with water to 10 ml, add 0.5 g of zinc powder, and heat ethanol (95 per cent) may be replaced by Industrial
on a water-bath for 5 minutes. To 2 ml of the clear supernatant Methylated Spirit, diluted so as to be of equivalent strength.
liquid add 0.5 ml of sodium nitrite solution, cool in ice and
add to 3 ml of 2-naphthol solution; an orange red colour is Benzalkonium Chloride Solution contains not less than
produced. 49.0 per cent w/v and not more than 51.0 per cent w/v of
alkylbenzyldimethylammonium chlorides, calculated as
C. To 25 mg add 1 ml of 2 M nitric acid; a white precipitate is C22H40ClN. It may contain not more than 16.0 per cent v/v of
produced which dissolves on addition of 5 ml of ethanol ethanol, C2H6O.
(95 per cent). The resulting solution gives reaction A of
chlorides (2.3.1). Description. A clear, colourless or slightly yellow, syrupy
liquid; odour, aromatic.
Tests
Identification
Acidity or alkalinity. Dissolve 0.5 g in 50 ml of carbon dioxide-
free water, add 0.1 ml of bromocresol purple solution and A. Dilute 0.2 ml with 10 ml of water. To 5 ml add 1.5 ml of dilute
titrate with 0.1 M hydrochloric acid or with 0.1 M sodium nitric acid; a white precipitate is produced which is soluble in
hydroxide. Not more than 0.1 ml is required to change the ethanol (95 per cent). To the remainder add 1.5 ml of mercuric
colour of the solution. chloride solution; a white precipitate is produced which is
soluble in ethanol (95 per cent).
Ammonia compounds. Boil 0.1 g with 3 ml of sodium hydroxide
solution; no odour of ammonia is produced. B. Evaporate 0.5 ml to dryness on a water-bath, dissolve the
residue in 1 ml of sulphuric acid, add 0.1 g of potassium
Foreign amines. Dissolve 0.1 g in 5 ml of water and add 3 ml nitrate, heat on a water-bath for 5 minutes, cool, dilute with
of 1 M sodium hydroxide; no precipitate is formed. Heat to water to 10 ml, add 0.5 g of zinc powder, and heat on a water-
boiling; the odour of amines is not perceptible. bath for 5 minutes. To 2 ml of the clear supernatant liquid add
Sulphated ash (2.3.18). Not more than 0.2 per cent. 0.5 ml of sodium nitrite solution, cool in ice and add to 3 ml of
2-naphthol solution; an orange red colour is produced.
Water (2.3.43). Not more than 10 per cent, determined on 0.3 g.
C. To 0.05 ml add 1 ml of 2 M nitric acid; a white precipitate is
Assay. Weigh accurately about 2.0 g, dissolve in sufficient produced which dissolves on addition of 5 ml of ethanol
water to produce 100.0 ml. Transfer 25.0 ml to a separating (95 per cent). The resulting solution gives reaction A of
funnel, add 25 ml of chloroform, 10 ml of 0.1 M sodium chlorides (2.3.1).
hydroxide and 10.0 ml of a freshly prepared 5 per cent w/v
solution of potassium iodide. Shake well, allow to separate Tests
and discard the chloroform layer. Shake the aqueous solution
Acidity or alkalinity. Dissolve 1.0 g in 50 ml of carbon dioxide-
with three further quantities, each of 10 ml, of chloroform and
free water, add 0.1 ml of bromocresol purple solution and
discard the chloroform layer. Add 40 ml of hydrochloric acid,
titrate with 0.1 M hydrochloric acid or with 0.1 M sodium
cool and titrate with 0.05 M potassium iodate until the solution
hydroxide. Not more than 0.1 ml is required to change the
becomes pale brown in colour. Add 2 ml of chloroform and
colour of the solution.
continue the titration until the chloroform becomes colourless.
Titrate a mixture of 20 ml of water, 10.0 ml of a freshly prepared Ammonia compounds. Boil 0.2 ml with 3 ml of sodium hydroxide
5 per cent w/v solution of potassium iodide and 40 ml of solution; no odour of ammonia is produced.
159
BENZATHINE PENICILLIN IP 2007
Foreign amines. To a volume containing 0.1 g of benzalkonium not less than 24.0 per cent and not more than 27.0 per cent of
chloride add sufficient water to produce 5 ml and add 3 ml of C16H20N2, both calculated on the anhydrous basis.
1 M sodium hydroxide; no precipitate is formed. Heat to Description. A white, crystalline powder; almost odourless.
boiling; the odour of amines is not perceptible.
Ethanol (if present) (2.3.45). Not more than 16.0 per cent v/v, Identification
determined by Method I or II, as applicable.
Sulphated ash (2.3.18). Not more than 0.2 per cent. B, C and D may be omitted if test A is carried out.
Assay. Weigh accurately about 4.0 g, dissolve in sufficient A. Determine by infrared absorption spectrophotometry (2.4.6).
water to produce 100.0 ml. Transfer 25.0 ml to a separating Compare the spectrum with that obtained with benzathine
funnel, add 25 ml of chloroform, 10 ml of 0.1 M sodium penicillin RS.
hydroxide and 10.0 ml of a freshly prepared 5 per cent w/v
solution of potassium iodide. Shake well, allow to separate B. Shake 0.1 g with 1 ml of 1 M sodium hydroxide for 2 minutes,
and discard the chloroform layer. Shake the aqueous solution add 2 ml of ether, shake for 1 minute and allow to separate.
with three further quantities, each of 10 ml, of chloroform and Evaporate 1 ml of the ether layer to dryness, dissolve the
discard the chloroform layer. Add 40 ml of hydrochloric acid, residue in 2 ml of glacial acetic acid and add 1 ml of potassium
cool and titrate with 0.05 M potassium iodate until the solution dichromate solution; a golden yellow precipitate is formed.
becomes pale brown in colour. Add 2 ml of chloroform and C. Shake 0.1 g with 2 ml of 1 M sodium hydroxide for 2 minutes,
continue the titration until the chloroform becomes colourless. extract the mixture with two quantities, each of 3 ml, of ether,
Titrate a mixture of 20 ml of water, 10.0 ml of a freshly prepared evaporate the combined extracts and dissolve the residue in
5 per cent w/v solution of potassium iodide and 40 ml of 1 ml of ethanol (50 per cent). Add 5 ml of picric acid solution,
hydrochloric acid with 0.05 M potassium iodate in a similar heat at 90° for 5 minutes and allow to cool slowly; the
manner; the difference between the titrations represents the precipitate, after recrystallisation from ethanol (25 per cent)
amount of 0.05 M potassium iodate required. containing a small quantity of picric acid, melts at about 214°
1 ml of 0.05 M potassium iodate is equivalent to 0.0354 g of (2.4.21).
C22H40ClN. Determine the relative density (2.4.29), and calculate D. In the Assay, the principal peak in the chromatogram
the amount of C22H40ClN, weight in volume. obtained with the test solution corresponds to the peak in the
Storage. Avoid contact with metals. chromatogram obtained with the reference solution.
Labelling. The label states, where appropriate, the content of Tests
ethanol (95 per cent) or Industrial Methylated Spirit.
pH (2.4.24). 5.0 to 7.5, determined in a saturated solution.
Benzathine Penicillin (2.4.14).
Benzathine Benzylpenicillin; Benzathine Penicillin G Prepare the solutions immediately before use. Avoid any
overheating during the preparation of the solutions.
Test solution. Dissolve an accurately weighed quantity of
H COO about 70 mg of the substance under examination in 25 ml of
NH2 O
H N CH3 methanol with the aid of ultrasound (for about 2 minutes).
N CH 3
Dilute to 50.0 ml with a solution containing 6.8 g per litre of
NH2 S
H H potassium dihydrogen phosphate and 1.02 g per litre of
O disodium hydrogen phosphate.
2
Reference solution (a). Dissolve an accurately weighed
quantity of about 70 mg of benzathine penicillin RS in 25 ml
C16H20N2,(C16H18N2O4S)2 Mol. Wt. 909.1 of methanol with the aid of ultrasound (for about 2 minutes).
Benzathine Penicillin is N,N’-dibenzylethylenediammonium Dilute to 50.0 ml with a solution containing 6.8 g per litre of
bis[(6R)-6-(2 phenylacetamido)penicillanate] containing a potassium dihydrogen phosphate and 1.02 g per litre of
variable amount of water. disodium hydrogen phosphate.
Benzathine Penicillin contains not less than 96.0 per cent and Reference solution (b).Dilute 1 ml of reference solution (a) to
not more than 100.5 per cent of C16H20N2,(C16H18N2O4S)2 and 100 ml with mobile phase A.
160
IP 2007 BENZATHINE PENICILLIN INJECTION
Chromatographic system C16H20N2,(C16H18N2O 4S)2 by multiplying the percentage

– a stainless steel column 25 cm x 4 mm, packed with content of benzylpenicillin by 1.36.
Benzathine Penicillin intended for use in the manufacture of
– mobile phase: A. a mixture of 10 volumes of a 34 g per
parenteral preparations without a further appropriate
litre solution of potassium dihydrogen phosphate
procedure for the removal of bacterial endotoxins complies
adjusted to pH 3.5 with phosphoric acid, 30 volumes of
with the following additional requirement.
methanol and 60 volumes of water,
B. a mixture of 10 volumes of a 34 g per Bacterial endotoxins (2.2.3). Not more than 0.13 Endotoxin
litre solution of potassium dihydrogen phosphate Unit per ml of a solution prepared in the following manner.
adjusted to pH 3.5 with phosphoric acid, 30 volumes of Suspend 20 mg of the substance under examination in 20 ml of
water and 60 volumes of methanol, 0.1 M sodium hydroxide, dilute 1 ml to 100 ml and use the
– temperature 40°, supernatant liquid.
– flow rate. 1 ml per minute, Sterility (2.2.11). Complies with the test for sterility.
below, Storage. Store protected from moisture at a temperature not
– spectrophotometer set at 220 nm, exceeding 30°. If the material is intended for use in the
– a 20 µl loop injector. manufacture of parenteral preparations the container should
be sterile and sealed so as to exclude micro-organisms.
Time Mobile phase A Mobile phase B
(min) (per cent v/v) (per cent v/v) Labelling The label states whether or not the contents are
0 – 10 75 25 intended for use in the manufacture of parenteral preparations.
10 – 20 75 → 0 25 →100
20 – 55 0 100
55 – 70 75 25 Benzathine Penicillin Injection
Inject reference solution (a). Relative retention time with
reference to benzylpenicillin: benzathine = 0.3 to 0.4; Benzathine Benzylpenicillin Injection; Benzathine
benzylpenicilloic acids benzathide = about 2.4. If necessary, Penicillin G Injection
adjust the concentration of methanol in the mobile phase. Benzathine Penicillin Injection is a sterile material consisting
Inject the test solution and reference solution (b). The area of of Benzathine Penicillin with or without suspending agents,
any secondary peak obtained with the test solution buffering agents and other excipients. It is filled in a sealed
corresponding to benzylpenicilloic acids benzathide is not container.
more than twice the sum of the areas of the two principal The injection is constituted by suspending the contents of
peaks in the chromatogram obtained with reference solution the sealed container in the requisite amount of sterile Water
(b) (2 per cent).The area any other secondary peak obtained for Injections, immediately before use.
with the test solution is not more than the sum of the areas of
the two principal peaks in the chromatogram obtained with Storage. The constituted suspension should be used
reference solution (b) (1 per cent). Disregard any peak with an immediately after preparation but, in any case, within the period
area 0.05 times the sum of the areas of the two principal peaks recommended by the manufacturer.
in the chromatogram obtained with reference solution (b) (0.05 Benzathine Penicillin Injection contains not less than 95.0 per
per cent). cent and not more than 110.0 per cent of the stated amount of
Water (2.3.43). 5.0 to 8.0 per cent, determined on 0.3 g. benzathine penicillin, C16H30N2, (C16H18N2O4S)2.
Assay. Determine by liquid chromatography, (2.4.14) as given Description. A white crystalline powder, almost odourless.
under the test for Related substances using the following
mobile phase.
Mobile phase. a mixture of 10 volumes of phosphate buffer (Powders for Injection) and with the following requirements.
solution pH 3.5, 35 volumes of methanol, and 55 volumes of
water. Identification
Inject alternately the test solution and reference solution (a). A. Shake 0.1 g with 1 ml of 1 M sodium hydroxide for 2 minutes,
Calculate the percentage contents of C 16H 20N 2 and of add 2 ml of ether, shake for 1 minute and allow to separate.
C 16 H 20 N 2 ,(C 16 H 18 N 2 O 4 S) 2 . Calculate the content of Evaporate 1 ml of the ether layer to dryness, dissolve the
161
BENZATHINE PENICILLIN INJECTION IP 2007
residue in 2 ml of glacial acetic acid and add 1 ml of potassium – temperature 40°,

dichromate solution; a golden yellow precipitate is formed. – flow rate. 1 ml per minute,
B. Shake 0.1 g with 2 ml of 1 M sodium hydroxide for 2 minutes, – a linear gradient programme using the conditions given
extract the mixture with two quantities, each of 3 ml, of ether, below,
evaporate the combined extracts and dissolve the residue in – spectrophotometer set at 220 nm,
1 ml of ethanol (50 per cent). Add 5 ml of picric acid solution, – a 20 µl loop injector.
heat at 90° for 5 minutes and allow to cool slowly; the Time Mobile phase A Mobile phase B
precipitate, after recrystallisation from ethanol (25 per cent) (min) (per cent v/v) (per cent v/v)
containing a small quantity of picric acid, melts at about 214°
(2.4.21). 0 – 10 75 25
10 – 20 75 → 0 25 →100
C. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak in the 20 – 55 0 100
chromatogram obtained with the reference solution. 55 – 70 75 25
Tests Inject reference solution (a). Relative retention time with
reference to benzylpenicillin: benzathine = 0.3 to 0.4;
pH (2.4.24). 5.0 to 7.5, determined in a suspension obtained by benzylpenicilloic acids benzathide = about 2.4. If necessary,
reconstituting as directed on the label. adjust the concentration of methanol in the mobile phase.
Consistency. To a quantity containing 60,000 Units add 2 ml Inject the test solution and reference solution (b). The area of
of water and shake thoroughly. The resulting suspension any secondary peak obtained with the test solution
passes through a 23G hypodermic needle. corresponding to benzylpenicilloic acids benzathide is not
Related substances. Determine by liquid chromatography more than twice the sum of the areas of the two principal
(2.4.14). peaks in the chromatogram obtained with reference solution
(b) (2 per cent).The area any other secondary peak obtained
Prepare the solutions immediately before use. Avoid any with the test solution is not more than the sum of the areas of
overheating during the preparation of the solutions. the two principal peaks in the chromatogram obtained with
Test solution. Dissolve an accurately weighed quantity reference solution (b) (1 per cent). Disregard any peak with an
containing about 70 mg of Benzathine Penicillin in 25 ml of area 0.05 times the sum of the areas of the two principal peaks
methanol with the aid of ultrasound (for about 2 minutes). in the chromatogram obtained with reference solution (b)
Dilute to 50.0 ml with a solution containing 6.8 g per litre of (0.05 per cent).
potassium dihydrogen phosphate and 1.02 g per litre of Bacterial endotoxins (2.2.3) Not more than 0.13 Endotoxin
disodium hydrogen phosphate. Unit per ml of a solution prepared by suspending 20 mg of the
Reference solution (a). Dissolve an accurately weighed substance under examination in 20 ml of 0.1 M sodium
quantity of about 70 mg of benzathine penicillin RS in 25 ml hydroxide, diluting 1 ml to 100 ml and using the supernatant.
of methanol with the aid of ultrasound (for about 2 minutes). Water (2.3.43). 5.0 to 8.0 per cent, determined on 0.3 g.
Dilute to 50.0 ml with a solution containing 6.8 g per litre of
potassium dihydrogen phosphate and 1.02 g per litre of Assay. Determine by liquid chromatography, (2.4.14) as given
disodium hydrogen phosphate. under the test for Related substances using the following
mobile phase.
Reference solution (b).Dilute 1 ml of reference solution (a) to
100 ml with mobile phase A. Mobile phase. a mixture of 10 volumes of phosphate buffer
water.
octadecylsilyl silica gel (5 µm), Inject alternately the test solution and reference solution (a).
Calculate the content of C 16 H 20 N 2 ,(C 16 H 18 N 2 O 4S) 2 by
multiplying the percentage content of benzylpenicillin by
adjusted to pH 3.5 with phosphoric acid, 30 volumes of
1.36.
B. a mixture of 10 volumes of a 34 g per Labelling. The label states (1) the directions for constituting
litre solution of potassium dihydrogen phosphate the suspension; (2) the names of any added buffering agents
adjusted to pH 3.5 with phosphoric acid, 30 volumes of or other pharmaceutical aids; (3) that the preparation is meant
water and 60 volumes of methanol, for intramuscular injection only.
162
IP 2007 FORTIFIED BENZATHINE PENICILLIN INJECTION
Fortified Benzathine Penicillin Consistency. To a quantity containing 600,000 Units of

Benzathine Penicillin, 300,000 Units each of Procaine Penicillin
Injection and Benzylpenicillin add 2 ml of water and shake thoroughly.
Fortified Benzathine Benzylpenicillin Injection; The resulting suspension passes readily through a 22G
Fortified Benzathine Penicillin G Injection hypodermic needle.
Benzathine Penicillin Injection is a sterile material consisting
Unit per ml of a solution prepared by suspending 20 mg of the
of Benzathine Penicillin and Procaine Penicillin with or without
substance under examination in 20 ml of 0.1 M sodium
suspending agents, buffering agents and other excipients. It
hydroxide, diluting 1 ml to 100 ml and using the supernatant.
is filled in a sealed container.
The injection is constituted by suspending the contents of Water (2.3.43). Not more than 7.5 per cent, determined on
the sealed container in the requisite amount of sterile Water 0.3 g.
for Injections containing Benzylpenicillin Sodium immediately Assay. For benzathine penicillin — Shake a quantity of the
before use. mixed contents of 10 containers containing 1 g of Benzathine
Storage. The constituted suspension should be used Penicillin with 30 ml of a saturated solution of sodium chloride
immediately after preparation but, in any case, within the period and 10 ml of 5 M sodium hydroxide and extract with four
and under the conditions recommended by the manufacturer. successive quantities, each of 50 ml, of ether. Wash the
combined ether extracts with three successive quantities, each
Fortified Benzathine Penicillin Injection contains not less than
of 5 ml, of water, extracting each aqueous washing with the
90.0 per cent and not more than 125.0 per cent of the stated
same 25 ml of ether. Combine the ether extracts, evaporate to
amount of benzathine penicillin, not less than 95.0 per cent
a low bulk, add 2 ml of ethanol and evaporate to dryness.
Dissolve the residue in 50 ml of glacial acetic acid and titrate
procaine penicillin, not less than 90.0 per cent and not more
with 0.1 M perchloric acid using 1 ml of 1-naphtholbenzein
than 130.0 per cent of the stated amount of benzylpenicillin
sodium, all in terms of Units of penicillin.
C18H50N6O8S2. Calculate the apparent content of Benzathine
Penicillin.
Calculate the content of procaine penicillin, as determined by
Identification the method given below in the weight of the sample used in
A. Give the reaction for penicillins (2.3.1). this assay, multiply this content by a factor of 1.544 and deduct
the figure from the apparent content of benzathine penicillin;
B. Give reaction B of penicillins and cephalosporins (2.3.1). the result is the content of benzathine penicillin. (1 mg of
C. Shake 0.1 g with 1 ml of 1 M sodium hydroxide for 2 minutes, benzathine penicillin is approximately equivalent to 1330 Units
add 2 ml of ether, shake for 1 minute and allow to separate. of penicillin).
Evaporate 1 ml of the ether layer to dryness, dissolve the For procaine penicillin — To a quantity of the mixed contents
residue in 2 ml of glacial acetic acid and add 1 ml of potassium of 10 containers containing 0.25 g of Procaine Penicillin add
dichromate solution; a golden yellow precipitate is formed. 100 ml of water, shake well, dilute to 200.0 ml with water, mix
D. Give the reactions of sodium salts (2.3.1). and filter. Dilute 5.0 ml of the filtrate to 250.0 ml with buffer
solution pH 7.0 and measure the absorbance of the resulting
Tests solution at the maximum at about 290 nm, using buffer solution
Stability. Using an aseptic technique prepare the suspension pH 7.0 as the blank (2.4.7). Calculate the content of procaine
as directed on the label in an individual unopened container penicillin taking 310 as the specific absorbance at 290 nm.
and determine the concentration of benzylpenicillin sodium (1 mg of procaine penicillin is equivalent to 1009 Units of
by the method described below using an accurately measured penicillin).
quantity of the suspension, withdrawn aseptically from the For benzylpenicillin sodium — Shake a quantity of the mixed
container. Store the remainder of the suspension in the closed contents of 10 containers containing 0.15 g of Benzylpenicillin
container at 4° for 7 days and then repeat the determination of Sodium with water until dissolved and dilute to 500.0 ml with
benzylpenicillin sodium. water. Dilute 25.0 ml of the resulting solution to 100.0 ml with
The concentration of benzylpenicillin sodium in the stored phosphate buffer pH 6.8. Place two quantities, each of 2.0 ml,
injection is not less than 80 per cent of the concentration of the resulting solution in separate stoppered tubes. To one
found in the freshly prepared suspension. tube add 10.0 ml of imidazole-mercury reagent, mix, stopper
163
BENZATHINE PENICILLIN TABLETS IP 2007
the tube and immerse in a water-bath at 60° for 35 minutes, Tests

swirling occasionally. Remove from the water-bath and cool
rapidly to 20° (solution A). Add 10.0 ml of imidazole solution Water (2.3.43). Not more than 8.0 per cent, determined on the
to the second tube, mix, stopper the tube and allow to stand at powdered tablets.
20° for 35 minutes, swirling occasionally (solution B). Without Other tests. Comply with the tests stated under Tablets.
delay measure the absorbance of solutions A and B at about
325 nm (2.4.7), using as the blank a mixture of 2.0 ml of water
(2.4.14).
and 10.0 ml of imidazole-mercury reagent for solution A and
a mixture of 2.0 ml of water and 10.0 ml of imidazole solution Prepare the solutions immediately before use. Avoid any
for solution B. Calculate the content of total penicillins as overheating during the preparation of the solutions.
C16H17N2NaO4S from the difference between the absorbances Test solution. Weigh and powder 20 tablets. Dissolve an
of solutions A and B, from the difference obtained by repeating accurately weighed quantity containing about 70 mg of
the procedure using 0.15 g of benzylpenicillin sodium RS in Benzathine Penicillin in 25 ml of methanol with the aid of
place of the contents of the sealed containers. Calculate the ultrasound (for about 2 minutes) and allow to stand for
content of benzylpenicillin sodium by subtracting the contents 15 minutes. Dilute to 50.0 ml with a solution containing 6.8 g
of benzathine penicillin and procaine penicillin, both expressed per litre of potassium dihydrogen phosphate and 1.02 g per
as benzylpenicillin sodium, C16H17N2NaO4S. litre of disodium hydrogen phosphate mix and filter.
Labelling. The label on the sealed container states (1) the Reference solution (a). Dissolve an accurately weighed
quantity of Benzathine Penicillin, Benzylpenicillin Sodium and quantity of about 70 mg of benzathine penicillin RS in 25 ml
Procaine Penicillin contained in it; (2) the directions for of methanol with the aid of ultrasound (for about 2 minutes).
reconstituting the suspension; (3) the names of the added Dilute to 50.0 ml with a solution containing 6.8 g per litre of
suspending agent, buffering agent and any other potassium dihydrogen phosphate and 1.02 g per litre of
pharmaceutical aid; (4) that the preparation is intended for disodium hydrogen phosphate.
intramuscular injection only.
Reference solution (b).Dilute 1 ml of reference solution (a) to
100 ml with mobile phase A.
Benzathine Penicillin Tablets Chromatographic system
Benzathine Benzylpenicillin Tablets; Benzathine octadecylsilyl silica gel (5 µm),
Penicillin G Tablets – temperature 40°,
Benzathine Penicillin Tablets contain Benzathine Penicillin – mobile phase: A. a mixture of 10 volumes of a 34 g per
equivalent to not less than 90.0 per cent and not more than litre solution of potassium dihydrogen phosphate
110.0 per cent of the stated number of Units of penicillin. adjusted to pH 3.5 with phosphoric acid, 30 volumes of
Identification B. a mixture of 10 volumes of a 34 g per
A. Shake 0.1 g with 1 ml of 1 M sodium hydroxide for 2 minutes, adjusted to pH 3.5 with phosphoric acid, 30 volumes of
add 2 ml of ether, shake for 1 minute and allow to separate. water and 60 volumes of methanol,
Evaporate 1 ml of the ether layer to dryness, dissolve the – flow rate. 1 ml per minute,
residue in 2 ml of glacial acetic acid and add 1 ml of potassium – a linear gradient programme using the conditions given
dichromate solution; a golden yellow precipitate is formed. below,
B. Shake 0.1 g with 2 ml of 1 M sodium hydroxide for 2 minutes, – spectrophotometer set at 220 nm,
extract the mixture with two quantities, each of 3 ml, of ether, – a 20 µl loop injector.
evaporate the combined extracts and dissolve the residue in Time Mobile phase A Mobile phase B
1 ml of ethanol (50 per cent). Add 5 ml of picric acid solution, (min) (per cent v/v) (per cent v/v)
heat at 90° for 5 minutes and allow to cool slowly; the 0 – 10 75 25
precipitate, after recrystallisation from ethanol (25 per cent)
containing a small quantity of picric acid, melts at about 214° 10 – 20 75 → 0 25 → 100
(2.4.21). 20 – 55 0 100
C. In the Assay, the principal peak in the chromatogram 55 – 70 75 25
obtained with the test solution corresponds to the peak in the Inject reference solution (a). Relative retention time with
chromatogram obtained with the reference solution. reference to benzylpenicillin: benzathine = 0.3 to 0.4;
164
IP 2007 BENZHEXOL TABLETS
benzylpenicilloic acids benzathide = about 2.4. If necessary, hydrochloride RS or with the reference spectrum of benzhexol
adjust the concentration of methanol in the mobile phase. hydrochloride.
Inject the test solution and reference solution (b). The area of B. Dissolve 0.5 g in 5 ml of warm methanol and make just
any secondary peak obtained with the test solution alkaline to litmus paper with 5 M sodium hydroxide; a
corresponding to benzylpenicilloic acids benzathide is not precipitate is produced, which, after recrystallisation from
more than twice the sum of the areas of the two principal methanol melts at about 114° (2.4.21).
peaks in the chromatogram obtained with reference solution
C. Gives the reactions of chlorides (2.3.1).
(b) (2 per cent). The area any other secondary peak obtained
with the test solution is not more than the sum of the areas of Tests
the two principal peaks in the chromatogram obtained with
reference solution (b) (1 per cent). Disregard any peak with an pH (2.4.24). 5.2 to 6.2, determined in a solution prepared by
area 0.05 times the sum of the areas of the two principal peaks dissolving 1.0 g in 50 ml of carbon dioxide-free water with the
in the chromatogram obtained with reference solution (b) aid of heat, cooling and diluting to 100.0 ml with the same
(0.05 per cent). solvent.
Assay. Determine by liquid chromatography, (2.4.14) as given Piperidylpropiophenone. Dissolve 0.1 g in a mixture of 40 ml
under the test for Related substances using the following of water and 1 ml of 1 M hydrochloric acid with the aid of
mobile phase. heat, cool and add sufficient water to produce 100.0 ml. The
absorbance of the resulting solution at about 247 nm is not
Mobile phase: a mixture of 10 volumes of?phosphate buffer
more than 0.5 (2.4.7).
water. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Inject alternately the test solution and reference solution (a). Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Calculate the percentage content of C16H20N2,(C16H18N2O4S)2
by multiplying the percentage content of benzylpenicillin by Assay. Weigh accurately about 0.7 g and dissolve in 50 ml of
1.36. anhydrous glacial acetic acid previously neutralised using
Storage. Store at a temperature not exceeding 30°. 1-naphtholbenzein solution as indicator, warming and cooling,
if necessary. Add 15 ml of mercuric acetate solution. Titrate
with 0.1 M perchloric acid to the full colour change of the
indicator. Carry out a blank titration.
Benzhexol Hydrochloride
Trihexyphenidyl Hydrochloride C20H31NO,HCl.
N
,HCl Benzhexol Tablets
OH Benzhexol HydrochlorideTablets; Trihexyphenidyl
Hydrochloride Tablets
C20H31NO,HCl Mol. Wt. 337.9 Benzhexol Tablets contain not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of benzhexol
Benzhexol Hydrochloride is (RS)-1-cyclohexyl-1-phenyl-3- hydrochloride, C20H31NO,HCl.
piperidinopropan-1-ol hydrochloride.
Benzhexol Hydrochloride contains not less than 98.0 per cent Identification
and not more than 101.0 per cent of C20H31NO,HCl, calculated A. Shake a quantity of the powdered tablets with 20 ml of
on the dried basis. water and filter. The filtrate yields a yellow precipitate with
Description. A white or creamy-white, crystalline powder; trinitrophenol solution and a white precipitate with 5 M
odourless or almost odourless. sodium hydroxide.
Identification the plate with silica gel G.
A. Determine by infrared absorption spectrophotometry (2.4.6). Mobile phase. A mixture of 90 volumes of chloroform and
Compare the spectrum with that obtained with benzhexol 10 volumes of methanol.
165
BENZOCAINE IP 2007
Test solution. Shake a quantity of the powdered tablets with Carry out the chromatographic procedure described under
sufficient chloroform to produce a solution containing 0.2 per Uniformity of content.
cent w/v of Benzhexol Hydrochloride and filter. Calculate the content of C20H31NO,HCl in the tablets.
Reference solution. A 0.2 per cent w/v solution of benzhexol
hydrochloride RS in chloroform.
Apply to the plate 10 µl of each solution. After development Benzocaine
remove the plate, allow it to dry in air and spray with dilute
potassium iodobismuthate solution. The principal spot in the
chromatogram obtained with the test solution corresponds to COOC2H5
that in the chromatogram obtained with the reference solution.
Tablets.
Determine by liquid chromatography (2.4.14). NH2
Test solution. Disperse well one tablet in 5.0 ml of water in an C9H11NO2 Mol. Wt. 165.2
ultrasonic bath, add 10 ml of methanol, shake for 15 minutes, Benzocaine is ethyl 4-aminobenzoate.
dilute to 25.0 ml with methanol, mix and filter through a filter
with a maximum pore size of 0.2 mm. Benzocaine contains not less than 99.0 per cent and not more
of benzhexol hydrochloride RS and 0.004 per cent w/v of Description. Colourless crystals or a white, crystalline powder;
3-piperidylpropiophenone hydrochloride RS in the mobile odourless.
phase.
Identification
– a stainless steel column 15 cm x 3.9 mm, packed with Test A may be omitted if tests B, C and D are carried out. Tests
octadecylsilyl silica gel (5 µm) (such as Resolve C18), B, C and D may be omitted if test A is carried out.
– mobile phase: 800 volumes of acetonitrile, 200 volumes A. Determine by infrared absorption spectrophotometry (2.4.6).
of water and 0.2 volume of triethylamine, the pH of the Compare the spectrum with that obtained with benzocaine RS
mixture being adjusted to 4.0 with phosphoric acid, or with the reference spectrum of benzocaine.
– spectrophotometer set at 210 nm, B. Dissolve 10 mg in 1 ml of water with the aid of one drop of
– a 20 µl loop injector. dilute hydrochloric acid and add 2 drops of a 10 per cent
w/v solution of sodium nitrite and 2 drops of a solution of 10
mg of 2-naphthol in 5 ml of sodium hydroxide solution; a
resolution factor between the two principal peaks in the
deep red colour is produced. On setting aside the solution for
chromatogram obtained with the reference solution is greater
some time, a scarlet precipitate is produced.
than 4.0.
C. Dissolve 0.2 g in 10 ml of water with the aid of dilute
hydrochloric acid (solution A) and divide into 2 parts. To one
Calculate the content of C20H31NO,HCl in the tablet. part of solution A add iodine solution; a precipitate is obtained
Other tests. Comply with the tests stated under Tablets. (distinction from orthocaine).
Assay. Weigh and powder 20 tablets. Determine by liquid D. To the other part of solution A add potassium mercuri-
chromatography (2.4.14), using the following solutions. iodide solution; no precipitate is obtained (distinction from
procaine).
Test Solution. Disperse well a quantity of the powdered tablets
containing about 5 mg of Benzhexol Hydrochloride in 5.0 ml of Tests
water in an ultrasonic bath, add 10 ml of methanol, shake for
15 minutes, dilute to 25.0 ml with methanol mix and filter Appearance of solution. A 5.0 per cent w/v solution in ethanol
through a filter with a maximum pore size of 0.2 µm. (95 per cent) is clear (2.4.1), and colourless (2.4.1).
Reference solution. A solution containing 0.02 per cent w/v of Acidity or alkalinity. Dissolve 0.5 g in 5 ml of ethanol (95 per
benzhexol hydrochloride RS and 0.01 per cent w/v of 3- cent), add 10 ml of water and one drop of phenolphthalein
piperidylpropiophenone hydrochloride RS in the mobile solution; no pink colour is produced. Add 0.5 ml of 0.01 M
phase. sodium hydroxide; the solution develops a pink colour.
166
IP 2007 COMPOUND BENZOIC ACID OINTMENT
Heavy metals (2.3.13). 2.0 g complies with the limit test for dissolve the cooled residue in 16 ml of brominated
heavy metals, Method B (10 ppm). hydrochloric acid and 45 ml of water. Remove the excess of
Chlorides. Dissolve 0.2 g in 5 ml of ethanol (95 per cent) bromine with 2 ml of stannous chloride AsT. The resulting
previously acidified with a few drops of dilute nitric acid and solution complies with the limit test for arsenic (2 ppm).
add few drops of silver nitrate solution; no turbidity is Heavy metals. Not more than 10 ppm, determined by the
produced immediately. following method. Dissolve 2.0 g in 25 ml of acetone and add
2 ml of water and 10 ml of hydrogen sulphide solution; any
colour produced is not more intense than that of a solution
Loss on drying (2.4.19). Not more than 0.5 per cent, determined prepared with 25 ml of acetone, 2.0 ml of lead standard solution
on 1.0 g by drying over phosphorus pentoxide at a pressure (10 ppm Pb) and 10 ml of hydrogen sulphide solution.
of 1.5 to 2.5 kPa.
Readily oxidisable substances. Add 1 ml of sulphuric acid to
Assay. Weigh accurately about 0.4 g and dissolve in a mixture 100 ml of water, heat to boiling and add dropwise 0.1 M
of 25 ml of hydrochloric acid and 50 ml of water. Cool to 10°. potassium permanganate until the pink colour persists for
Determine by the nitrite titration (2.3.31). 30 seconds. Dissolve exactly 1 g in the hot solution and titrate
1 ml of 0.1 M sodium nitrite is equivalent to 0.01652 g of with 0.1 M potassium permanganate to a pink colour that
C9H11NO2. persists for 15 seconds; not more than 0.5 ml of 0.1 M
potassium permanganate is required.
Readily carbonisable substances. Dissolve 0.5 g in 5 ml of
sulphuric acid and allow to stand for 5 minutes. The colour of
the solution is not more intense than that of reference solution
Benzoic Acid YS5 (2.4.1).
COOH Cinnamic acid. Warm 0.1 g with 0.1 g of potassium
permanganate and 5 ml of dilute sulphuric acid; no odour of
benzaldehyde is developed.
Chlorinated compounds. Dissolve 0.33 g in 5 ml of 0.5 M
sodium carbonate, evaporate to dryness and heat the residue
C7H6O2 Mol. Wt. 122.1 until completely charred, keeping the temperature below 400°.
Benzoic Acid contains not less than 99.5 per cent and not Extract the residue with a mixture of 10 ml of water and 12 ml of
more than 100.5 per cent of C7H6O2, calculated on the dilute nitric acid and filter; the filtrate complies with the limit
anhydrous basis. test for chlorides (2.3.12).
Description. Colourless, light crystals, scales or needles; Sulphated ash (2.3.18). Not more than 0.1 per cent.
odour, slight and characteristic. Water (2.3.43). Not more than 0.7 per cent, determined on
0.25 g and using a mixture of 1 volume of methanol and
Identification 2 volumes of pyridine as the solvent.
A. Warm gently 0.2 g with 20 ml of water, add 1 ml of 1 M Assay. Weigh accurately about 1.0 g and dissolve in 15 ml of
sodium hydroxide and filter. To the filtrate add ferric chloride warm ethanol (95 per cent) previously neutralised to
test solution; a buff coloured precipitate is produced. phenolphthalein solution. Add 20 ml of water and titrate with
B. When examined in the range 220 nm to 360 nm, a 0.001 per 0.5 M sodium hydroxide using phenolphthalein solution as
cent w/v solution in methanol shows an absorption maximum indicator.
only at about 225 nm; absorbance at about 225 nm, about 0.8 1 ml of 0.5 M sodium hydroxide is equivalent to 0.06106 g of
(2.4.7). C7H6O2.
C. A 1 per cent w/v solution is acid to methyl red solution.
Tests Compound Benzoic Acid Ointment

Appearance of solution. A 5.0 per cent w/v solution in ethanol Benzoic and Salicylic Acids Ointment; Whitfield’s
(95 per cent) is clear (2.4.1), and colourless (2.4.1). Ointment
Arsenic (2.3.10). Mix 5.0 g with 3 g of anhydrous sodium Compound Benzoic Acid Ointment is an ointment containing
carbonate, add 10 ml of bromine solution and mix thoroughly. 6.0 per cent w/w of Benzoic Acid and 3.0 per cent w/w of
Evaporate to dryness on a water-bath, gently ignite and Salicylic Acid in a suitable ointment base. Other strengths
167
BENZOIN IP 2007
may also be prepared with Benzoic Acid and Salicylic Acid extract with the same 50 ml of ether. Combine the aqueous
being in the ratio of about 2 to 1. extracts, cautiously add hydrochloric acid until the solution
is distinctly acid to litmus paper and extract with 4 quantities,
Compound Benzoic Acid Ointment contains not less than
each of 25 ml, of ether; combine the extracts and evaporate
5.7 per cent and not more than 6.3 per cent w/w of benzoic
the ether at a temperature below 40°. Dissolve the residue in
acid, C7H6O2, and not less than 2.85 per cent and not more
5 ml of 0.5 M sodium hydroxide, add 50.0 ml of 0.1 M bromine
than 3.15 per cent w/w of salicylic acid, C7H6O3.
and 5 ml of hydrochloric acid, shake repeatedly during
15 minutes and allow to stand for 15 minutes. Add 10 ml of
Identification potassium iodide solution and titrate with 0.1 M sodium
Carry out the method for thin-layer chromatography (2.4.17), thiosulphate using starch solution, added towards the end
coating the plate with silica gel GF254. of the titration, as indicator. Repeat the operation without the
substance under examination. The difference between the
Mobile phase. A mixture of 80 volumes of toluene and titrations represents the amount of bromine required.
20 volumes of glacial acetic acid.
1 ml of 0.1 M bromine is equivalent to 0.002302 g of C7H6O3.
Test solution. Warm 1 g of the ointment with 10 ml of chloroform,
Storage. Store at a temperature not exceeding 30°
cool and filter.
benzoic acid and 0.3 per cent w/v of salicylic acid in
chloroform. Benzoin
Apply to the plate 2 µl of each solution. After development, Benzoin is the balsamic resin obtained from Styrax benzoin
dry the plate in a current of air and examine in ultraviolet light Dryander or Styrax paralleloneurus Perkins, known in
at 254 nm. The two principal spots in the chromatogram commerce as Sumatra Benzoin or from Styrax tonkinensis
obtained with the test solution correspond to those in the (Pierre) Craib ex Hartwich, or other species of the Section
chromatogram obtained with the reference solution. Examine Anthostyrax of the genus Styrax, known in commerce as Siam
the plate in ultraviolet light at 365 nm. A blue fluorescent spot Benzoin (Fam. Styraceae).
in the chromatogram obtained with the test solution
corresponds in colour and position to the one in the Benzoin contains not less than 25.0 per cent of total balsamic
chromatogram obtained with the reference solution. Spray acids, calculated as cinnamic acid, C9H8O2, in Sumatra Benzoin
the plate with ferric chloride test-solution. The chromatogram and as benzoic acid, C7H6O2, in Siam Benzoin, calculated on
obtained with the test solution shows a purple spot the dried basis.
corresponding in position to the blue fluorescent spot Description. Unground Sumatra Benzoin — Blocks or lumps
observed in ultraviolet light at 365 nm and corresponding in of varying size, made up of tears compacted together, with a
colour and position to the spot in the chromatogram obtained reddish-brown, reddish-grey or greyish-brown resinous mass,
with the reference solution. known in commerce as block benzoin. It also occurs in the
form of tears with cream-coloured to yellowish surfaces; when
Tests fractured they exhibit milky-white surfaces; odour, balsamic
which accentuates on digestion with boiling water.
Assay. For benzoic acid — Weigh accurately about 2.5 g,
dissolve with the aid of gentle heat, as completely as possible, Unground Siam Benzoin — Pebble-like tears of variable size
in 50 ml of a mixture of equal volumes of ethanol (95 per cent) and shape, compressed, yellowish-brown to rusty-brown
and ether, previously neutralised to phenolphthalein solution externally, milky white on fracture, hard and brittle at ordinary
and titrate with 0.1 M sodium hydroxide using temperatures but softened by heat; odour, balsamic.
phenolphthalein solution as indicator.
Identification
1 ml of 0.1 M sodium hydroxide, after deducting 1 ml for each
0.01381 g of C7H6O3 in the weight of the ointment taken A. To a solution in ethanol (95 per cent) add water; the
(calculated from the result of the Assay for salicylic acid) is solution becomes milky, and the mixture is acid to litmus paper.
equivalent to 0.01221 g of C7H6O2. B. Heat 0.5 g in a dry test-tube; it melts and evolves white
For salicylic acid — Weigh accurately about 2.5 g, dissolve fumes, which form a white needle-shaped crystalline sublimate.
with the aid of gentle heat, as completely as possible, in C. Heat 0.5 g in a test-tube with 5 ml of potassium
50 ml of ether, and extract with 5 quantities, each of 10 ml, of a permanganate solution; a strong odour of benzaldehyde is
saturated solution of sodium bicarbonate, washing each obtained with Sumatra Benzoin.
168
IP 2007 BENZOIN
D. Determine by thin-layer chromatography (2.4.17), coating Benzoin, determined by the following method. Weigh
the plate with kieselguhr. accurately about 2 g, in coarse powder, in a tared extraction
thimble and insert the thimble in a Soxhlet or other suitable
Mobile phase. A mixture of 93 volumes of toluene and
continuous extraction apparatus. Place 0.1 g of sodium
7 volumes of ethyl acetate.
hydroxide in the receiving flask of the apparatus, extract with
Test solution. Dissolve 2.0 g of the substance under ethanol (95 per cent) until extraction is complete (about
examination in 100 ml of ethanol (95 per cent). 5 hours), dry the thimble to constant weight at 105° and
calculate the ethanol-soluble extractive from the increase in
Reference solution (a). A 0.05 per cent w/v solution of benzoic
weight of the thimble.
acid RS in chloroform.
Acid-insoluble ash (2.3.19). Not more than 1.0 per cent in
Sumatra Benzoin and not more than 0.5 per cent in Siam
cinnamic acid RS in chloroform.
Benzoin, determined on 2.0 g.
Reference solution (c). A 0.05 per cent w/v solution of coniferyl
Loss on drying (2.4.19). Not more than 10.0 per cent,
benzoate RS in chloroform. determined on 2.0 g, in coarse powder, by drying over
Reference solution (d). A 0.05 per cent w/v solution of phosphorus pentoxide at a pressure not exceeding 2.7 kPa for
cinnamoyl cinnamate RS in chloroform. 4 hours.
Reference solution (e). A 0.05 per cent w/v solution of propyl Assay. Weigh accurately about 1.25 g and boil with 25 ml
cinnamate RS in chloroform. of dilute ethanolic potassium hydroxide solution under a
reflux condenser for 1 hour. Remove the ethanol and digest
Reference solution (f). A 0.05 per cent w/v solution of the residue with 50 ml of hot water until diffused. Cool the
cinnamoyl benzoate RS in chloroform. liquid, add 150 ml of water and 1.5 g of magnesium
After development, dry the plate in air until the odour of the sulphate dissolved in 50 ml of water. Mix thoroughly and
solvent is no longer detectable and spray with anisaldehyde- set aside for 10 minutes. Filter, wash the residue on the
sulphuric acid reagent. Heat the plate at 110° for 5 minutes filter with 20 ml of water, acidify the combined filtrate and
and examine in ultraviolet light at 254 nm. In the case of Sumatra washings with hydrochloric acid and extract with
Benzoin, the chromatogram obtained with the test solution successive quantities of 50, 40, 30, 30 and 30 ml of ether.
exhibits four intense spots corresponding to spots in the Combine the ether extracts and discard the aqueous portion.
chromatograms obtained with reference solutions (b), (c), (d) Extract with successive quantities of 20, 20, 10, 10 and 10
and (e). In the case of Siam Benzoin, it exhibits intense spots ml of sodium bicarbonate solution, washing each aqueous
corresponding to spots in the chromatograms obtained with extract with the same 20 ml of ether. Discard the ether
reference solutions (a), (c), (d) and (f). layers, acidify the combined aqueous extracts with
hydrochloric acid and extract with successive quantities
Tests of 30, 20, 20 and 10 ml of chloroform, filtering each
chloroform extract through a plug of cotton wool on which
Dammar gum. Determine by thin-layer chromatography a layer of anhydrous sodium sulphate is placed. Evaporate
(2.4.17), coating the plate with aluminium oxide G. the chloroform on a water-bath until about 10 ml remains
and remove the remainder in a current of air stopping
Mobile phase. A mixture of 60 volumes of ether and 40 volumes
immediately when the last trace of solvent is removed.
of light petroleum (80° to 100° ).
Dissolve the residue by warming with 10 ml of ethanol (95
Test solution. Dissolve by warming 0.2 g of the substance per cent), previously neutralised to phenol red solution,
under examination in 10 ml of ethanol (90 per cent) and cool and titrate with 0.1 M sodium hydroxide using phenol
centrifuge. red solution as indicator.
Apply to the plate 5 µl of the test solution. Allow the mobile 1 ml of 0.1 M sodium hydroxide is equivalent to 0.01482 g of
phase to rise 10 cm. Dry the plate in air, spray with total balsamic acids, calculated as cinnamic acid, C9H8O2, in
anisaldehyde-sulphuric acid reagent and heat at 100° to 105° Sumatra Benzoin and 0.01221 g of total balsamic acids,
for 5 minutes. The chromatogram does not show any calculated as benzoic acid, C7H6O2, in Siam Benzoin.
prominent spot with an Rf value between 0.4 and 1.0.
Storage. Store protected from light at a temperature not
Foreign organic matter (2.6.1). Not more than 1.0 per cent. exceeding 30°.
Ethanol-soluble extractive. Not less than 75.0 per cent in Labelling. The label states whether the material is Sumatra
Sumatra Benzoin and not less than 90.0 per cent in Siam Benzoin or Siam Benzoin.
169
COMPOUND BENZOIN TINCTURE IP 2007
Compound Benzoin Tincture to spots in the chromatograms obtained with reference

solutions (a), (c), (d) and (f).
Friars’ Balsam
Benzoin, in moderately coarse powder 100 g the plate with kieselguhr.
Prepared Storax 75 g Mobile phase. A mixture of 93 volumes of toluene and
Tolu Balsam 25 g 7 volumes of ethyl acetate.
Aloes, in moderately coarse powder 20 g Test solution. Dilute 1 ml of the tincture with 4 ml of ethanol
Ethanol (90 per cent) sufficient to (95 per cent).
produce 1000 ml Reference solution. A 0.05 per cent w/v solution of styrene RS
Macerate the Benzoin, Prepared Storax, Tolu Balsam and Aloes in chloroform.
with 800 ml of Ethanol (90 per cent) in a closed vessel for not
less than 2 days with occasional shaking. Filter and pass
dry the plate in air until the odour of the solvent is no longer
sufficient Ethanol (90 per cent) through the filter to produce
detectable and spray with anisaldehyde-sulphuric acid
the required volume.
reagent. Heat the plate at 110° for 5 minutes and examine in
Compound Benzoin Tincture contains not less than 4.5 per ultraviolet light at 254 nm. The chromatogram obtained with
cent w/v of total balsamic acids, calculated as cinnamic acid, the test solution exhibits an intense spot corresponding to
C9H8O2. the spot in the chromatogram obtained with the reference
solution (Prepared Storax).
Identification
A. Determine by thin-layer chromatography (2.4.17), coating the plate with kieselguhr.
the plate with kieselguhr.
Mobile phase. A mixture of 93 volumes of toluene and 7 volumes of ethyl acetate.
Test solution. Dilute 1 ml of the tincture with 4 ml of ethanol
Test solution. Dilute 1 ml of the tincture with 4 ml of ethanol (95 per cent).
(95 per cent). (The chromatographic profile may vary
depending on the variety of Benzoin used). Reference solution (a). A 0.05 per cent w/v solution of benzoyl
benzoate RS in chloroform.
Reference solution (a). A 0.05 per cent w/v solution of benzoic
acid RS in chloroform. Reference solution (b). A 0.05 per cent w/v solution of benzoyl
cinnamate RS in chloroform.
cinnamic acid RS in chloroform. Reference solution (c). A 0.05 per cent w/v solution of eugenol
RS in chloroform.
Reference solution (c). A 0.05 per cent w/v solution of coniferyl
benzoate RS in chloroform. Reference solution (d). A 0.05 per cent w/v solution of vanillin
RS in chloroform.
Reference solution (d). A 0.05 per cent w/v solution of
cinnamoyl cinnamate RS in chloroform. Apply to the plate 20 µl of each solution. After development,
Reference solution (e). A 0.05 per cent w/v solution of propyl detectable and spray with anisaldehyde-sulphuric acid
cinnamate RS in chloroform. reagent. Heat the plate at 110° for 5 minutes and examine in
Reference solution (f). A 0.05 per cent w/v solution of ultraviolet light at 254 nm. The chromatogram obtained with
cinnamoyl benzoate RS in chloroform. the test solution exhibits spots corresponding to the spots in
Apply to the plate 20 µl of each solution. After development, the chromatogram obtained with reference solutions, (a), (b),
dry the plate in air until the odour of the solvent is no longer (c) and (d) (Tolu Balsam).
detectable and spray with anisaldehyde-sulphuric acid D. Carry out the method for thin-layer chromatography (2.4.17),
reagent. Heat the plate at 110° for 5 minutes and examine in coating the plate with silica gel G.
ultraviolet light at 254 nm. In the case of Sumatra Benzoin, the
Mobile phase. A mixture of 100 volumes of ethyl acetate,
chromatogram obtained with the test solution exhibits four
13.5 volumes of methanol and 10 volumes of water.
intense spots corresponding to spots in the chromatograms
obtained with reference solutions (b), (c), (d) and (e). In the Test solution. Dilute 1 ml of the tincture with 4 ml of ethanol
case of Siam Benzoin, it exhibits intense spots corresponding (95 per cent).
170
IP 2007 BENZYL ALCOHOL
Reference solution. A 0.5 per cent w/v solution of barbaloin Labelling. The label states that it is flammable.
RS in methanol.
Apply to the plate 50 µl of each solution as bands 20 mm long
and not more than 3 mm wide. Allow the mobile phase to rise Benzyl Alcohol
10 cm. Dry the plate in air until the odour of the solvent is no
longer detectable, spray with a 10 per cent w/v solution of C7H8O Mol. Wt. 108.1
potassium hydroxide in methanol and examine in ultraviolet Benzyl AIcohol contains not less than 97.0 per cent of C7H8O.
light at 365 nm. The chromatogram obtained with the test
solution exhibits a yellow fluorescent band corresponding to Description. A colourless liquid; almost odourless; taste,
the band obtained in the chromatogram obtained with the sharp and burning.
reference solution and a light blue fluorescent band with a
lower Rf value due to aloesine. Heat the plate at 110° for Identification
5 minutes; a violet fluorescent band just below the band Add three drops to a strong potassium permanganate
corresponding to barbaloin may also be seen in the solution, acidified with sulphuric acid; benzaldehyde,
chromatogram obtained with the test solution (Aloes). recognizable by its odour, is produced.
Tests
Tests
Weight per ml (2.4.29). 0.870 g to 0.885 g.
Wt. per ml (2.4.29). 1.64 g to 1.05 g.
Ethanol content. 70.0 to 77.0 per cent v/v, determined by
Method II (2.3.45). Distillation range (2.4.8). None distils below 200° and not
less than 94 per cent distils between 202° and 208°.
Total solids. Not less than 13.5 per cent w/v, determined on
1 ml by drying in an oven at 105° for 4 hours. Refractive index (2.4.27). 1.536 to 1.542.
Assay. Evaporate 10 ml to a thick consistency on a water- Acid Value (2.3.23). Not more than 0.5.
bath. Boil the residue with 25 ml of ethanolic potassium Chlorinated compounds. Mix 2.0 g with 50 ml of amyl alcohol
hydroxide solution under a reflux condenser for 1 hour. in a dry flask, add in small quantities 3 g of sodium, connect
Remove the ethanol and digest the residue with 50 ml of hot the flask to a reflux air condenser, warm gently until the
water until diffused. Cool the liquid, add 150 ml of water and evolution of hydrogen ceases, and boil gently for one hour.
1.5 g of magnesium sulphate dissolved in 50 ml of water. Mix Cool the liquid to a little below 100° add 50 ml of water, 5.0 ml
thoroughly and set aside for 10 minutes. Filter, wash the residue of 0.1M silver nitrate, and 20 ml of nitric acid, and titrate the
on the filter with 20 ml of water, acidify the combined filtrate excess of silver nitrate with 0.1 M ammonium thiocyanate,
and washings with hydrochloric acid and extract with using ferric ammonium sulphate solution as indicator. Repeat
successive quantities of 50, 40, 30, 30 and 30 ml of ether. the operation without the sample; the difference between the
Combine the ether extracts and discard the aqueous portion. titrations does not exceed 0.3 ml.
Extract with successive quantities of 20, 20, 10, 10 and 10 ml of
sodium bicarbonate solution, washing each aqueous extract Benzaldehyde. Mix in a stoppered cylinder 10 ml with 10 ml of
with the same 20 ml of ether. Discard the ether layers, acidify aldehyde-free alcohol and 20 ml of hydroxylamine
the combined aqueous extracts with hydrochloric acid and hydrochloride solution. Allow to stand for five minutes and
extract with successive quantities of 30, 20, 20 and 10 ml of titrate with 0.1 M sodium hydroxide to the same green colour
chloroform, filtering each chloroform extract through a plug as that shown by 20 ml of hydroxylamine hydrochloride
of cotton wool on which a layer of anhydrous sodium sulphate solution contained in a similar cylinder, both solutions being
is placed. Evaporate the chloroform on a water-bath until about viewed down the axes of the cylinders; not more than 1.4 ml of
10 ml remains and remove the remainder in a current of air 0.1 M sodium hydroxide is required.
stopping immediately when the last trace of solvent is Assay. To 1.5 g add 25 ml of a mixture of 1 volume of acetic
removed. Dissolve the residue by warming with 10 ml of anhydride and 7 volumes of pyridine and heat on a water-
ethanol (95 per cent), previously neutralised to phenol red bath for thirty minutes. Cool, add 25 ml of water, and titrate
solution, cool and titrate with 0.1 M sodium hydroxide using with 1 M sodium hydroxide, using phenolphthalein solution
phenol red solution as indicator. as indicator. Repeat the operation without the substance under
1 ml of 0.1 M sodium hydroxide is equivalent to 0.01482 g of examination; the difference between the titrations represents
total balsamic acids, calculated as cinnamic acid, C9H8O2. the amount of alkali required by the benzyl alcohol.
Storage. Store protected from light in tightly-closed containers 1 ml of 1 M sodium hydroxide is equivalent to 0.1081 g of
and avoid exposure to direct sunlight and to excessive heat. C7H8O.
171
BENZYL BENZOATE IP 2007
Storage. Store protected from moisture in a container with using 0.2 ml of phenolphthalein solution as indicator. Add
minimum space above the level of the liquid. 40 ml of 0.5 M ethanolic potassium hydroxide and boil under
a reflux condenser on a water-bath for 1 hour. Add 20 ml of
water and titrate the excess of alkali with 0.5 M hydrochloric
Benzyl Benzoate acid using a further 0.2 ml of phenolphthalein solution as
indicator. Repeat the operation without the substance under
O examination. The difference between the titrations represents
the alkali required to saponify the benzyl benzoate.
O
1 ml of 0.5 M ethanolic potassium hydroxide is equivalent to
0.1061 g of C14H12O2.
C14H12O2 Mol. Wt. 212.6 Storage. Store protected from light and air in well-filled
containers.
Benzyl Benzoate is the benzyl ester of benzoic acid.
Benzyl Benzoate contains not less than 99.0 per cent and not
more than 100.5 per cent w/w of C14H12O2.
Benzyl Benzoate Application
Description. Colourless crystals or a clear, colourless, oily
Benzyl Benzoate Application contains 25 per cent w/w of
liquid; odour, faintly aromatic.
Benzyl Benzoate in a suitable oil-in-water emulsified basis.
Identification Benzyl Benzoate Application contains not less than 22.5 per
A. Boil 2 g with 25 ml of ethanolic potassium hydroxide cent and not more than 27.5 per cent w/w of benzyl benzoate,
solution for 2 hours in a flask fitted with a reflux condenser. C14H12O2.
Remove the ethanol on a water-bath, add 50 ml of water to the Assay. Weigh accurately about 8.0 g and dissolve in 10 ml of
liquid remaining in the flask and distil until the liquid distilling ethanol (95 per cent) previously neutralised with 0.1 M
is no longer turbid. Preserve the distillate for test B. To the sodium hydroxide contained in a hard-glass flask and
liquid remaining in the flask add dilute hydrochloric acid till neutralise the free acid in the solution with 0.5 M ethanolic
it is neutral and divide the mixture into two parts. To one part potassium hydroxide using 0.2 ml of phenolphthalein solution
add ferric chloride test solution; a buff-coloured precipitate as indicator. Add 40 ml of 0.5 M ethanolic potassium
is produced. To the other part add hydrochloric acid; a white hydroxide and boil under a reflux condenser on a water-bath
crystalline precipitate of benzoic acid is produced. for 1 hour. Add 20 ml of water and titrate the excess of alkali
B. To the distillate obtained in test A, add 2.5 g of potassium with 0.5 M hydrochloric acid using a further 0.2 ml of
permanganate and 2 ml of sodium hydroxide solution, boil phenolphthalein solution as indicator. Repeat the operation
for 15 minutes in a flask fitted with a reflux condenser, cool without the substance under examination. The difference
and filter. To the filtrate add dilute hydrochloric acid till it is between the titrations represents the alkali required to saponify
neutral and divide the mixture into two parts. To one part add the benzyl benzoate.
ferric chloride test solution; a buff-coloured precipitate is 1 ml of 0.5 M ethanolic potassium hydroxide is equivalent to
produced. To the other part add hydrochloric acid; a white 0.1061 g of C14H12O2.
crystalline precipitate of benzoic acid is produced.
Labelling. The label states that the contents should be shaken
Tests before use.
Congealing temperature (2.4.10). Not below 17.0°.

Relative density (2.4.29). 1.113 g to 1.118 g. Benzylpenicillin Potassium
Refractive index (2.4.27). 1.567 to 1.569. Penicillin G Potassium
Sulphated ash (2.3.18). Not more than 0.1 per cent. H COOK
O
Assay. Boil a convenient quantity of ethanol (95 per cent) N CH3
thoroughly to expel carbon dioxide and neutralise to H
N S CH3
phenolphthalein solution. Weigh accurately about 2 g of the
substance under examination, dissolve in 5 ml of the neutralised H H
O
ethanol contained in a hard-glass flask and neutralise the free
acid in the solution with 0.5 M ethanolic potassium hydroxide C16H17KN2O4S Mol. Wt. 372.5
172
IP 2007 BENZYLPENICILLIN POTASSIUM
Benzylpenicillin Potassium is potassium (6R)-6-(2- Prepare the solutions immediately before use.
phenylacetamido)penicillanate, produced by the growth of Test solution (a). Dissolve 50.0 mg of the substance under
certain strains of Penicillium notatum or related organisms, examination in water and dilute to 50.0 ml with the same solvent.
or obtained by any other means.
Test solution (b). Dissolve 80.0 mg of the substance under
Benzylpenicillin Potassium contains not less than 96.0 per
examination in water and dilute to 20.0 ml with the same solvent.
cent and not more than 100.5 per cent of penicillins, calculated
as C16H17KN2O4S on the dried basis. Reference solution (a). Dissolve 50.0 mg of benzylpenicillin
sodium RS in water and dilute to 50.0 ml with the same solvent.
Description. A white or almost white, crystalline powder.
Reference solution (b). Dissolve 10 mg of benzylpenicillin
Identification sodium RS and 10 mg of phenylacetic acid RS in water and
dilute to 50.0 ml with the same solvent.
Reference solution (c). Dilute 1.0 ml of reference solution (a)
Compare the spectrum with that obtained with benzylpenicillin
to 20.0 ml with water. Dilute 1.0 ml of the solution to 50.0 ml
potassium RS.
with the same solvent.
B. Gives reaction A of potassium salts (2.3.1).
Reference solution (d). Dilute 4.0 ml of reference solution (a)
Tests to 100.0 ml with water.
Specific optical rotation (2.4.22). +270° to +300°, determined octadecylsilyl silica gel (5 µm),
in a 2.0 per cent w/v solution in carbon dioxide-free water. – mobile phase: A. a mixture of 10 volumes of a 68 g per
Light absorption (2.4.7). Dissolve 94 mg in sufficient water to litre solution of potassium dihydrogen phosphate
produce 50.0 ml. Measure the absorbance of the solution at adjusted to pH 3.5 with a 500 g per litre solution of
about 325 nm, at about 280 nm and at the maximum at about dilute phosphoric acid, 30 volumes of methanol and
264 nm, diluting the solution, if necessary, for the measurement 60 volumes of water,
at the maximum at about 264 nm. Absorbances at about B. a mixture of 10 volumes of a 68 g per
325 nm and 280 nm, not more than 0.10 and that at the maximum litre solution of potassium dihydrogen phosphate
at about 264 nm, calculated on the basis of the undiluted adjusted to pH 3.5 with a 500 g per litre solution of
solution (0.188 per cent w/v), 0.80 to 0.88. dilute phosphoric acid, 40 volumes of water and
50 volumes of methanol,
Related substances. Determine by liquid chromatography – flow rate. 1 ml per minute,
(2.4.14) as given under Assay. Inject reference solution (d) – spectrophotometer set at 225 nm,
and elute isocratically using the chosen mobile phase. Inject – a 20 µl loop injector.
test solution (b) and start the elution isocratically. Immediately
after elution of the benzylpenicillin peak start the following Equilibrate the column with a mobile phase ratio A:B of 70:30.
linear gradient: Inject reference solution (b). The test is not valid unless the
Time Mobile Mobile Comment resolution between the two principal peaks is at least 6.0
phase A phase B (if necessary, adjust the ratio A:B of the mobile phase) and the
(min) (per cent v/v) (per cent v/v) capacity factor for the second peak (benzylpenicillin) is 4.0 to
6.0.
0 – 20 70 → 0 30 → 100 linear gradient
Inject reference solution (c). Adjust the system to obtain a
20 – 35 0 100 Isocratic peak with a signal-to-noise ratio of at least 3.
35 – 50 70 30 re-equilibration Inject alternately test solution (a) and reference solution (a).
Inject water and use the same elution pattern to obtain a
blank. In the chromatogram obtained with test solution (b) the Calculate the percentage content of benzylpenicillin potassium
area of any peak, other than the principal peak, is not greater by multiplying the percentage content of benzylpenicillin
than the area of the principal peak in the chromatogram sodium by 1.045.
obtained with reference solution (d) (1.0 per cent). Benzylpenicillin Potassium intended for use in the
manufacture of parenteral preparations without a further
appropriate procedure for the removal of bacterial
endotoxins complies with the following additional
Assay. Determine by liquid chromatography (2.4.14). requirement.
173
BENZYLPENICILLIN SODIUM IP 2007
Bacterial endotoxins (2.2.3). Not more than 0.16 Endotoxin Related substances. Determine by liquid chromatography
Unit per mg. (2.4.14) as given under Assay. Inject reference solution (d)
Benzylpenicillin Potassium intended for use in the and elute isocratically using the chosen mobile phase. Inject
manufacture of parenteral preparations without a further test solution (b) and start the elution isocratically. Immediately
appropriate sterilisation procedure complies with the after elution of the benzylpenicillin peak start the following
following additional requirement. linear gradient:
Time Mobile Mobile Comment
Sterility (2.2.11). Complies with the test for sterility.
phase A phase B
Storage. Store protected from moisture at a temperature not (min) (per cent v/v) (per cent v/v)
exceeding 30°. If it is intended for use in the manufacture of 0 – 20 70 → 0 30 → 100 linear gradient
parenteral preparations, the container should be sterile and
sealed so as to exclude micro-organisms. 20 – 35 0 100 Isocratic
35 – 50 70 30 re-equilibration
Labelling. The label states whether or not the contents are
intended for use in the manufacture of parenteral preparations. Inject water and use the same elution pattern to obtain a
blank. In the chromatogram obtained with test solution (b) the
area of any peak, other than the principal peak, is not greater
than the area of the principal peak in the chromatogram
obtained with reference solution (d) (1.0 per cent).
Benzylpenicillin Sodium
Penicillin G Sodium on 1.0 g by drying in an oven at 105°.
C16H17N2NaO4S Mol. Wt. 356.4 Assay. Determine by liquid chromatography, (2.4.14).
Benzylpenicillin Sodium is sodium (6R)-6-(2- Prepare the solutions immediately before use.
phenylacetamido) penicillanate, produced by the growth of Test solution (a). Dissolve 50.0 mg of the substance under
certain strains of Penicillium notatum or related organisms, examination in water and dilute to 50.0 ml with the same solvent.
or obtained by any other means.
Teat solution (b). Dissolve 80.0 mg of the substance under
Benzylpenicillin Sodium contains not less than 96.0 per cent examination in water and dilute to 20.0 ml with the same solvent.
and not more than 100.5 per cent of penicillins, calculated as
C16H17N2NaO4S on the dried basis. Reference solution (a). Dissolve 50.0 mg of benzylpenicillin
sodium RS in water and dilute to 50.0 ml with the same solvent.
Identification sodium RS and 10 mg of phenylacetic acid RS in water and
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution (c). Dilute 1.0 ml of reference solution (a)
Compare the spectrum with that obtained with benzylpenicillin to 20.0 ml with water. Dilute 1.0 ml of the solution to 50.0 ml
sodium RS. with the same solvent.
B. Gives reaction A of sodium salts (2.3.1). Reference solution (d). Dilute 4.0 ml of reference solution (a)
to 100.0 ml with water.
Tests
pH (2.4.24). 5.5 to 7.5, determined in a 10.0 per cent w/v solution. – a stainless steel a column 25 cm x 4.6 mm, packed with
in a 2.0 per cent w/v solution in carbon dioxide-free water.
Light absorption (2.4.7). Dissolve 90 mg in sufficient water to adjusted to pH 3.5 with a 500 g per litre solution of
produce 50.0 ml. Measure the absorbance of the solution at dilute phosphoric acid, 30 volumes of methanol and
about 325 nm, at about 280 nm and at the maximum at about 60 volumes of water,
264 nm, diluting the solution, if necessary, for the measurement B. a mixture of 10 volumes of a 68 g per
at about 264 nm. Absorbances at about 325 nm and 280 nm, litre solution of potassium dihydrogen phosphate
not more than 0.10 and that at the maximum at about 264 nm, adjusted to pH 3.5 with a 500 g per litre solution of
calculated on the basis of the undiluted solution (0.18 per dilute phosphoric acid, 40 volumes of water and
cent w/v), 0.80 to 0.88. 50 volumes of methanol,
174
IP 2007 BENZYLPENICILLIN INJECTION
– flow rate. 1 ml per minute, Storage. The constituted solution should be used immediately
– spectrophotometer set at 225 nm, after preparation but, in any case, within the period
– a 20 µl loop injector. recommended by the manufacturer.
Equilibrate the column with a mobile phase ratio A:B of Benzylpenicillin Injection contains not less than 95.0 per cent
70:30. and not more than 110.0 per cent of the stated amount of
penicillins, calculated as C16H18N2O4S.
resolution between the two principal peaks is at least 6.0 Description. A white or almost white crystalline powder.
(if necessary, adjust the ratio A:B of the mobile phase) and the
capacity factor for the second peak (benzylpenicillin) is 4.0 to
6.0.
peak with a signal-to-noise ratio of at least 3. Identification
Inject alternately test solution (a) and reference solution (a). A. Determine by infrared absorption spectrophotometry (2.4.6).
Calculate the percentage content of C16H17N2NaO4S. Compare the spectrum with that obtained with benzylpenicillin
potassium RS or benzylpenicillin sodium RS.
Benzylpenicillin Sodium intended for use in the manufacture
of parenteral preparations without a further appropriate B. Gives reaction A of potassium or sodium salts (2.3.1).
procedure for the removal of bacterial endotoxins complies Tests
Bacterial endotoxins (2.2.3). Not more than 0.16 Endotoxin pH (2.4.24). 5.5 to 7.5, determined in a 10.0 per cent w/v solution.
Unit per mg. Related substances. Determine by liquid chromatography
Benzylpenicillin Sodium intended for use in the manufacture (2.4.14) as given under Assay. Inject reference solution (d)
of parenteral preparations without a further appropriate and elute isocratically using the chosen mobile phase. Inject
sterilisation procedure complies with the following test solution (b) and start the elution isocratically. Immediately
additional requirement. after elution of the benzylpenicillin peak start the following
linear gradient:
Storage. Store protected from moisture at a temperature not phase A phase B
exceeding 30°. If it is intended for use in the manufacture of (min) (per cent v/v) (per cent v/v)
parenteral preparations, the container should be sterile and 0 – 20 70 → 0 30 → 100 linear gradient
sealed so as to exclude micro-organisms.
20 – 35 0 100 Isocratic
35 – 50 70 30 re-equilibration
Inject water and use the same elution pattern to obtain a
blank. In the chromatogram obtained with test solution (b) the
Benzylpenicillin Injection obtained with reference solution (d) (1.0 per cent).
Penicillin G Injection Bacterial endotoxins (2.2.3). Not more than 0.16 Endotoxin
Benzylpenicillin Injection is a sterile material consisting of Unit per mg.
Benzylpenicillin Potassium or Benzylpenicillin Sodium with or Loss on drying (2.4.19). Not more than 1.0 per cent, determined
without buffering agents and other excipients. It is filled in a on 1.0 g by drying in an oven at 105°.
sealed container.
Assay. Determine by liquid chromatography, (2.4.14).
sealed container in the requisite amount of sterile Water for Prepare the solutions immediately before use.
Injections, immediately before use. Determine the weight of the contents of 10 containers.
The constituted solution complies with the requirements for Test solution (a). Dissolve 50.0 mg of the mixed contents of
Clarity of solution and Particulate matter stated under the 10 containers in water and dilute to 50.0 ml with the same
Parenteral Preparations (Injections). solvent.
175
BETAHISTINE HYDROCHLORIDE IP 2007
Teat solution (b). Dissolve 80.0 mg of the substance under Betahistine Hydrochloride
examination in water and dilute to 20.0 ml with the same solvent.
Reference solution (a). Dissolve 50.0 mg of benzylpenicillin H
sodium RS in water and dilute to 50.0 ml with the same solvent. N N
CH3 , 2HCl
sodium RS and 10 mg of phenylacetic acid RS in water and
dilute to 50.0 ml with the same solvent C8H12N2, 2HCl Mol. Wt. 209.12
Reference solution (c). Dilute 1.0 ml of reference solution (a) Betahistine Dihydrochloride is N-methyl-2-(2-pyridyl)
to 20.0 ml with water. Dilute 1.0 ml of the solution to 50.0 ml ethylamine dihydrochloride.
with water.
Betahistine Hydrochloride contains not less than 98.5 per cent
Reference solution (d). Dilute 4.0 ml of reference solution (a) and not more than 102.0 per cent of C8H14Cl2N2, 2HCl, calculated
to 100.0 ml with water. on the dried basis.
Chromatographic system Description. A white to off-white, crystalline powder;
– a stainless steel column 25 cm x 4.6 mm, packed with sometimes clumped, odourless or almost odourless, very
octadecylsilyl silica gel (5 µm), hygroscopic.
litre solution of potassium dihydrogen phosphate Identification
adjusted to pH 3.5 with a 500 g per litre solution of A. Determine by infrared absorption spectrophotometry (2.4.6).
dilute phosphoric acid, 30 volumes of methanol and Compare the spectrum with that obtained with betahistine
60 volumes of water, hydrochloride RS.
B. a mixture of 10 volumes of a 68 g per
litre solution of potassium dihydrogen phosphate B In the Assay, the principal peak in the chromatogram obtained
adjusted to pH 3.5 with a 500 g per litre solution of with the test solution corresponds to the peak in the
dilute phosphoric acid, 40 volumes of water and chromatogram obtained with the reference solution.
50 volumes of methanol, C. Gives the reaction A of chlorides (2.3.12).
Appearance of solution. A 10 per cent w/v solution in water is
Equilibrate the column with a mobile phase ratio A:B of 70:30. clear (2.4.1) and not more intensely coloured than reference
Inject reference solution (b). The test is not valid unless the solution B8 (2.4.1).
resolution between the two principal peaks is at least 6.0 pH (2.4.24). 2.0 to 3.0, determined in a 10 per cent w/v solution
(if necessary, adjust the ratio A:B of the mobile phase) and the in water.
capacity factor for the second peak (benzylpenicillin) is 4.0 to
6.0.
(2.4.14).
peak with a signal-to-noise ratio of at least 3.
examination in 100 ml of the mobile phase.
Inject alternately test solution (a) and reference solution (a).
Calculate the content of benzylpenicillin sodium in the betahistine hydrochloride RS in the mobile phase.
injection. 1mg of C16H17N2NaO4S is equivalent to 0.9383 mg of
C16H18N2O4S.
100 ml with the mobile phase.
exceeding 30°.
Labelling. The label states (1) whether the contents are octadecylsilane chemically bonded to porous silica
Benzylpenicillin Potassium or Benzylpenicillin Sodium; (2) the (5 µm),
name of any added buffering agents. – mobile phase: dissolve 0.45 g ammonium acetate and
0.4 ml glacial acetic acid in 650 ml of water, add 350 ml
of acetonitrile and add 2.88 g of sodium laurylsulphate
and mix,
176
IP 2007 BETAHISTINE TABLETS
– flow rate. 0.5 ml per minute, B. In the Assay, the principal peak in the chromatogram
– spectrophotometer set at 254 nm, obtained with the test solution corresponds to the peak in the
Inject reference solution (a). The test is not valid unless the Tests
tailing factor is not more than 2.0 and the column efficiency in
not less than 2000 theoretical plates. Dissolution (2.5.2).
Inject the test solution and reference solution (b). In the Apparatus. No 2
chromatogram obtained with the test solution, the area of any Medium. Phosphate citrate buffer pH 6.8.
secondary peak is not more than 0.5 times the area of the peak Speed and time. 50 rpm and 30 minutes.
in the chromatogram obtained with reference solution (b)
the medium if necessary, at the maximum at about 256 nm
(2.4.7). Calculate the content of C8H14Cl2N2,2HCl in the medium
Heavy metals (2.3.13). 1 g complies with the limit test for heavy from the absorbance obtained from a solution of known
metals, Method C ( 20 ppm). concentration of betahistine hydrochloride RS in the same
Sulphated ash (2.3.18). Not more than 0.1 per cent. medium.
D. Not less than 80 per cent of the stated amount of C8H14Cl2N2,
2HCl.
on 1 g by drying in an oven at 105º.
Assay. Determine by liquid chromatography (2.4.14). (2.4.14).
Test solution. Dissolve 40 mg of the substance under Test solution. Weigh and powder 20 tablets. Weigh accurately
examination in 100.0 ml of the mobile phase. a quantity of the powdered tablet containing 32 mg of
Reference solution. A 0.04 per cent w/v solution of Betahistine Dihydrochloride, disperse in 50 ml of mobile phase
betahistine hydrochloride RS in the mobile phase. and dilute to 100 ml with mobile phase and filter.
Chromatographic system as described under Related Reference solution (a). A 0.032 per cent w/v solution of
substances. betahistine hydrochloride RS in mobile phase.
than 2.0 per cent. Chromatographic system
octadecylsilyl silica (5 µm),
Calculate the content of C8H14Cl2N2,2HCl. – column temperature 50°,
Storage. Store protected from light. – mobile phase: dissolve 2.76 g of sodium dihydrogen
phosphate monohydrate and 1.60 g of sodium
dodecylsulphate in 600 ml of water, add 1.2 g of
hexylamine and 400 ml of acetonitrile, and mix, adjusted
pH to 3.5 with orthophosphoric acid,
Betahistine Tablets – flow rate. 1 ml per minute,
Betahistine Hydrochloride Tablets – spectrophotometer set at 254 nm,
Betahistine Tablets contain Betahistine Dihydrochloride.
Betahistine Tablets contain not less than 95.0 per cent and tailing factor is not more than 2.0 and the column efficiency in
not more than 105.0 per cent of the stated amount of not less than 2000 theoretical plates.
betahistine dihydrochloride, C8H14Cl2N2, 2HCl.
Identification chromatogram obtained with the test solution, the area of any
A. Determine by infrared absorption spectrophotometry (2.4.6). in the chromatogram obtained with reference solution (b)
Compare the spectrum with that obtained with betahistine (0.5 per cent) and the sum of areas of all the secondary peaks
hydrochloride RS. is not more than 1.5 times the area of the peak in the
177
BETAMETHASONE IP 2007
chromatogram obtained with the reference solution (b) Betamethasone is 9α-fluoro-11β,17α,21-trihydroxy-16β-

(1.5 per cent). methylpregna-1,4-diene-3,20-dione.
Uniformity of content. Comply with the tests stated under Betamethasone contains not less than 96.0 per cent and not
Tablets. more than 104.0 per cent of C22H29FO5, calculated on the dried
Determine by liquid chromatography (2.4.14), as described basis.
under Assay. Description. A white to creamy-white powder; odourless.
Test solution. Crush one tablet in 100 ml volumetric flask. Add
about 50 ml of mobile phase and swirl for 10 minutes, make up Identification
to volume with mobile phase and filter. Test A may be omitted if tests B, C and D are carried out. Tests
Calculate the content of C8H14Cl2N2,2HCl. C and D may be omitted if tests A and B are carried out.
Other tests. Comply with the tests stated under Tablets. A. Determine by infrared absorption spectrophotometry (2.4.6).
Assay. Determine by liquid chromatography (2.4.14). Compare the spectrum with that obtained with betamethasone
Test solution. Weigh and powder 20 tablets. Weigh accurately RS or with the reference spectrum of betamethasone.
a quantity of the powdered tablet containing 32 mg of B. Determine by thin-layer chromatography (2.4.17), coating
Betahistine Dihydrochloride, disperse in 50 ml of mobile phase the plate with a suitable silica gel containing a fluorescent
and dilute to 100.0 ml with mobile phase and filter. indicator with an optimal intensity at 254 nm (such as Merck
Reference solution. A 0.032 per cent w/v solution of silica gel 60 F254).
betahistine hydrochloride RS in mobile phase. Mobile phase. A mixture of 85 volumes of ether, 10 volumes of
Chromatographic system toluene and 5 volumes of 1-butanol saturated with water.
– a stainless steel column 25 cm x 4.6 mm packed with Test solution. Dissolve 25 mg of the substance under
octadecylsilyl silica (5 µm), examination in a mixture of 90 volumes of chloroform and
– column temperature 50º, 10 volume of methanol.
– mobile phase: dissolve 2.76 g of sodium dihydrogen
phosphate monohydrate and 1.6 g of sodium Reference solution (a). A 0.25 per cent w/v solution of
dodecylsulphate in 600 ml of water. add 0.4 g of betamethasone RS in a mixture of 90 volumes of chloroform
hexylamine and 400 ml of acetonitrile, adjust the pH to and 10 volumes of methanol.
3.5 with orthophosphoric acid, Reference solution (b). A 0.125 per cent w/v solution of each
– flow rate. 2 ml per minute, of the substance under examination and betamethasone RS in
– spectrophotometer set at 254 nm, the same solvent mixture.
Reference solution (c). A 0.125 per cent w/v solution of each
Inject the reference solution. The test is not valid unless the of the substance under examination and dexamethasone RS
tailing factor is not more than 2.0, the column efficiency in not in the same solvent mixture.
less than 2000 theoretical plates. The relative standard
deviation for replicate injections is not more than 2.0 per cent. Apply to the plate 2 µl of each solution. After development,
dry the plate in air and spray with ethanolic sulphuric acid
Inject the test solution and the reference solution. (20 per cent). Heat at 120° for 10 minutes or until spots are
Calculate the content of C8H14Cl2N2,2HCl. produced, allow to cool and examine in daylight and in
Storage. Store protected from light. ultraviolet light at 365 nm. The principal spot in the
chromatogram obtained with the test solution is similar in
colour in daylight, in fluorescence in ultraviolet light at
Betamethasone 365 nm, position and size to the principal spot in the
chromatogram obtained with reference solution (a) and the
O OH chromatogram obtained with reference solution (b) shows only
H3 C one spot. The test is not valid unless the chromatogram
HO OH obtained with reference solution (c) shows two principal spots
H3C H CH3 that are close to one another but separated.
F H C. Heat 0.5 ml of chromic-sulphuric acid in a test-tube (5 cm ×

about 6 mm) in a naked flame until white fumes are evolved;
O
the solution wets the sides of the tube readily and there is no
C22H29FO5 Mol. Wt. 392.5 greasiness. Add 2 or 3 mg of the substance under examination
178
IP 2007 BETAMETHASONE TABLETS
and again heat in a naked flame until white fumes appear; the Adjust the sensitivity of the system so that the height of the
solution does not wet the sides of the tube and does not pour principal peak in the chromatogram obtained with reference
easily from the tube. solution (b) is not less than 50 per cent of the full scale of the
D. Place 2 ml of a 0.01 per cent w/v solution in ethanol in a recorder.
stoppered tube, add 10 ml of phenylhydrazine solution, mix, Inject reference solution (a). When the chromatograms are
warm in a water-bath at 60° for 20 minutes and cool immediately; recorded in the conditions described above, the retention times
absorbance of the resulting solution at about 450 nm, not are: methylprednisolone, about 11.5 minutes and
more than 0.25, (2.4.7). betamethasone, about 12.5 minutes. The test is not valid unless
the resolution between the peaks corresponding to
Tests methylprednisolone and betamethasone is at least 1.5; if
necessary, adjust the concentration of acetonitrile in mobile
phase A.
Inject separately a mixture of equal volumes of acetonitrile
Light absorption (2.4.7). Absorbance of a 0.001 per cent w/v
and methanol as blank, the test solution and reference solution
solution in ethanol (95 per cent) at the maximum at about
(b). In the chromatogram obtained with the test solution: the
240 nm, 0.37 to 0.40.
Related substances. Determine by liquid chromatography than the area of the principal peak in the chromatogram
(2.4.14). obtained with reference solution (b) (1.0 per cent) and not
Test solution. Dissolve 25.0 mg of the substance under more than one such peak has an area greater than half the area
examination in a mixture of equal volumes of acetonitrile and of the principal peak in the chromatogram obtained with
methanol and dilute to 10.0 ml with the same solvent. reference solution (b) (0.5 per cent); the sum of the areas of all
the peaks, other than the principal peak, is not greater than
Reference solution (a). Dissolve 2 mg of betamethasone RS twice the area of the principal peak in the chromatogram
and 2 mg of methylprednisolone RS in mobile phase A and obtained with reference solution (b) (2.0 per cent). Disregard
dilute to 100.0 ml with the same mobile phase. any peak due to the blank and any peak with an area less than
Reference solution (b). Dilute 1.0 ml of the test solution to 0.05 times the area of the principal peak in the chromatogram
100.0 ml with mobile phase A. obtained with reference solution (b).
Chromatographic system Sulphated ash (2.3.18). Not more than 0.1 per cent.
– a stainless steel column 25 cm ξ 4.6 mm, packed with Loss on drying (2.4.19). Not more than 0.5 per cent, determined
octadecylsilyl silica gel (5 µm), on 1.0 g by drying at a pressure not exceeding 0.7 kPa for
temperature 45°, 3 hours.
– mobile phase: A. a mixture of 250 ml of acetonitrile and
700 ml of water, allowed to equilibrate, sufficient water Assay. Dissolve 0.1 g in alcohol and dilute to 100.0 ml with the
added to produce 1000 ml and mixed, same solvent. Dilute 2.0 ml of the solution to 100.0 ml with
B. acetonitrile, alcohol. Measure the absorbance of the resulting solution at
– flow rate. 2.5 ml per minute, the maximum at about 238.5 nm (2.4.7).
– a linear gradient programme using the conditions given Calculate the percentage content of C22H29FO5 taking 395 as
below, the specific absorbance at 238.5 nm.
phase A phase B
(min) (per cent v/v) (per cent v/v)
Betamethasone Tablets
0 – 15 100 → 0 0 isocratic Betamethasone Tablets contain not less than 90.0 per cent
15 – 40 0 → 100 100 linear gradient betamethasone, C22H29FO5.
41– 46 100 → 0 0 equilibration
Identification
Equilibrate the column with mobile phase B for at least
30 minutes and then with mobile phase A for 5 minutes. For Powder a few tablets and extract with chloroform. Evaporate
subsequent chromatograms, use the conditions described from the extract to dryness. The residue complies with the following
40 minutes to 46 minutes. tests.
179
BETAMETHASONE TABLETS IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). supernatant liquid into a glass-stoppered tube and
Compare the spectrum with that obtained with betamethasone evaporate the ethanol on a water-bath with the aid of a
RS or with the reference spectrum of betamethasone. current of air to about 0.5 ml, then evaporate without heat
to dryness. Pipette 1 ml of a mixture of 9 volumes of
B. Place 2 ml of a 0.01 per cent w/v solution in ethanol in a
chloroform and 1 volume of methanol, insert the stopper
stoppered tube, add 10 ml of phenylhydrazine solution, mix,
warm in a water-bath at 60° for 20 minutes and cool immediately; and mix. Centrifuge, if necessary, to remove any insoluble
absorbance of the resulting solution at about 450 nm, not material. Use this solution as the test solution.
more than 0.25 (2.4.7). Determine by thin-layer chromatography (2.4.17), coating the
C. Determine by thin-layer chromatography (2.4.17), coating plate with a suitable silica gel containing a fluorescent indicator
the plate with silica gel G. with an optimal intensity at 254 nm (such as Merck silica gel
60 F254).
Solvent mixture. A mixture of 90 volumes of acetone and
10 volumes of formamide. Mobile phase. A mixture of 77 volumes of dichloromethane,
15 volumes of ether, 8 volumes of methanol and 1.2 volumes
Mobile phase. Chloroform. of water.
Test solution. Dissolve 25 mg of the residue in 10 ml of the
Reference solution (a). A 0.02 per cent w/v solution of the
solvent mixture.
substance under examination in a mixture of 90 volumes of
Reference solution (a). Dissolve 25 mg of betamethasone RS chloroform and 10 volumes of methanol.
in 10 ml of the solvent mixture.
Reference solution (b). Mix equal volumes of the test solution substance under examination in the same solvent mixture.
and reference solution (a).
Reference solution (c). A 0.1 per cent w/v solution of each of
Reference solution (c). Mix equal volumes of the test solution the substance under examination and prednisone RS in the
and a 0.25 per cent w/v solution of dexamethasone RS in the same solvent mixture.
solvent mixture.
Place the dry plate in a tank containing a shallow layer of the dry the plate in air until the odour of solvents is no longer
solvent mixture, allow the solvent mixture to ascend to the detectable and examine in ultraviolet light at 254 nm. Any
top, remove the plate from the tank and allow the solvent to secondary spot in the chromatogram obtained with the test
evaporate. Use within 2 hours, with the flow of the mobile solution is not more intense than the spot in the chromatogram
phase in the direction in which the aforementioned treatment obtained with reference solution (a) and not more than one
was done. such spot is more intense than the spot in the chromatogram
Apply to the plate 2 µl of each solution. Allow the mobile obtained with reference solution (b). The test is not valid
phase to rise 12 cm. Dry the plate in a current of warm air, unless the chromatogram obtained with reference solution (c)
allow the solvent to evaporate, heat at 120° for 15 minutes shows two clearly separated principal spots.
and spray the hot plate with ethanolic sulphuric acid
(20 per cent v/v). Heat at 120° for a further 10 minutes, allow Dissolution (2.5.2).
to cool and examine in daylight and in ultraviolet light at Apparatus. No 1
365 nm. The principal spot in the chromatogram obtained
with the test solution corresponds to that in the chromatogram Medium. 900 ml of water and 1 ml of 0.05 per cent w/v solution
obtained with reference solution (a). The principal spot in the of testosterone RS (internal standard) in methanol.
chromatogram obtained with reference solution (b) appears Speed and time. 50 rpm and 45 minutes.
as a single, compact spot. The chromatogram obtained with
reference solution (c) shows two closely running spots. Use one tablet in the vessel for each test.
Withdraw a suitable volume of the medium and filter. Determine
Tests by liquid chromatography (2.4.14)
Related substances. Transfer a quantity of the powdered Test solution. The filtrate obtained as given above.
tablets containing about 2 mg of Betamethasone to a glass-
stoppered 50-ml centrifuge tube. Pipette 20 ml of ethanol Reference solution. Dilute a mixture of 1.0 ml each of a 0.05 per
(95 per cent) into the tube, shake for 2 minutes and allow cent w/v solution of betamethasone RS in methanol and 1 ml
to stand for 20 minutes with occasional shaking. Centrifuge of a 0.05 per cent w/v solution of testosterone RS in methanol
the mixture for 5 minutes. Pipette 10 ml of the clear to 900 ml with water.
180
IP 2007 BETAMETHASONE SODIUM PHOSPHATE
Chromatographic system Betamethasone Sodium Phosphate

ONa
– mobile phase: a mixture of 60 volumes of methanol and O O P O
40 volumes of water, H3C
– flow rate. 2 ml per minute, HO OH ONa
– spectrophotometer set at 254 nm, H3C H CH3
F H
D: Not less than 75 per cent of the stated amount of C22H29FO5.
O
Tablets. Determine by liquid chromatography (2.4.14). C22H28FNa2O8P Mol. Wt. 516.4
Test solution. Finely crush one tablet, add 20.0 ml of a 0.002 Betamethasone Sodium Phosphate is 9α-fluoro- 11β,17α,21-
per cent w/v solution of hydrocortisone (internal standard) trihydroxy-16β-methyl-pregna-1,4-diene-3,20-dione
in methanol (50 per cent), shake for 10 minutes and filter disodium phosphate.
through a glass-fibre filter paper.
Betamethasone Sodium Phosphate contains not less than 96.0
Reference solution. A solution containing 0.0025 per cent w/v per cent and not more than 103.0 per cent of C22H28FNa2O8P,
of betamethasone RS and 0.002 per cent w/v of calculated on the anhydrous basis.
hydrocortisone.
Description. A white or almost white powder; odourless; very
NOTE — Protect the solutions from light. hygroscopic.
Identification
octadecylsilyl silica gel (5 µm), A. To 2 ml of a 0.013 per cent w/v solution in ethanol (95 per
– mobile phase: a mixture of 53 volumes of water and 47 cent) in a stoppered tube add 10 ml of phenylhydrazine-
volumes of methanol. sulphuric acid solution, mix, warm in a water-bath at 60° for
– flow rate. 1.4 ml per minute, 20 minutes and cool immediately. Absorbance of the resulting
– spectrophotometer set at 238 nm, solution at the maximum at about 450 nm, not more than 0.13
– a 20 µl loop injector. (2.4.7).
Calculate the content of C22H29FO5 in the tablet. B. Determine by thin-layer chromatography (2.4.17), coating
Mobile phase. A freshly prepared mixture of 30 volumes of
Assay. Determine by liquid chromatography (2.4.14). isopropyl alcohol, 10 volumes of acetic acid and 10 volumes
Test solution. Weigh and powder 20 tablets. Weigh accurately of water.
a quantity of the powder containing 2.5 mg of Betamethasone, Test solution. Dissolve 0.25 g of the substance under
add 20.0 ml of methanol (50 per cent), shake for 10 minutes examination in 100 ml of water.
and filter through a glass-fibre paper.
Reference solution (a). A solution containing 0.0125 per cent betamethasone sodium phosphate RS.
w/v of betamethasone RS and 0.01 per cent w/v of
hydrocortisone RS (internal standard). Reference solution (b). A mixture of equal volumes of the test
Reference solution (b). Prepare in the same manner as the test
solution but use 20.0 ml of a 0.01 per cent w/v solution of Reference solution (c). A mixture of equal volumes of the test
hydrocortisone in methanol (50 per cent) in place of 20.0 ml solution and a 0.25 per cent w/v solution of prednisolone
of methanol (50 per cent). sodium phosphate RS.
NOTE — Protect the solutions from light. Apply to the plate 2 µl of each solution. After development,
dry the plate in air until the odour of solvents is no longer
Carry out the chromatographic procedure described under detectable, spray with ethanolic sulphuric acid (20 per cent),
Uniformity of content. Calculate the content of C22H29FO5 in heat at 120° for 10 minutes, allow to cool, and examine in
the tablets. ultraviolet light at 365 nm. The principal spot in the
Storage. Store protected from light. chromatogram obtained with the test solution corresponds to
181
BETAMETHASONE EYE DROPS IP 2007
that in the chromatogram obtained with reference solution (a). Reference solution (a). A 1.0 per cent w/v solution of
The principal spot in the chromatogram obtained with betamethasone sodium phosphate RS in methanol.
reference solution (b) appears as a single, compact spot and Reference solution (b). A 0.02 per cent w/v solution of
the chromatogram obtained with reference solution (c) shows betamethasone RS in methanol.
two closely running spots.
C. Heat 0.5 ml of chromic-sulphuric acid in a test-tube (5 cm ´
dry the plate in air for 5 minutes and examine in ultraviolet
about 6 mm) in a naked flame until white fumes are evolved;
light at 254 nm. Any spot in the chromatogram obtained with
the solution wets the sides of the tube readily and there is no
the test solution other than that corresponding to
greasiness. Add 2 or 3 mg of the substance under examination
betamethasone sodium phosphate RS is not more intense
and again heat in a naked flame until white fumes appear; the
solution does not wet the sides of the tube and does not pour
solution (b).
easily from the tube.
D. Dissolve 2 mg in 2 ml of sulphuric acid and allow to stand Water (2.3.43). Not more than 8.0 per cent, determined on
for 5 minutes; no red colour or yellowish-green fluorescence 0.5 g.
is produced (distinction from prednisolone sodium phosphate Assay. Weigh accurately about 0.2 g and dissolve in sufficient
and hydrocortisone sodium phosphate). water to produce 200.0 ml. Dilute 5.0 ml to 250.0 ml with water
E. Heat gently 40 mg with 2 ml of sulphuric acid until white and measure the absorbance of the resulting solution at the
fumes are evolved, add nitric acid dropwise until oxidation is maximum at about 241 nm (2.4.7). Calculate the content of
complete and cool. Add 2 ml of water, heat until white fumes C22H28FNa2O8P, taking 297 as the specific absorbance at
are again evolved, cool, add 10 ml of water and neutralise to 241 nm.
litmus paper with dilute ammonia solution. The solution gives Storage. Store protected from light and moisture.
the reactions of sodium salts and of phosphates (2.3.1).
Tests
Betamethasone Eye Drops
Appearance of solution. A 2.0 per cent w/v solution is clear
(2.4.1), and colourless (2.4.1). Betamethasone Eye Drops are a sterile solution of
Betamethasone Sodium Phosphate in Purified Water.
Betamethasone Eye Drops contain not less than 90.0 per cent
Specific optical rotation (2.4.22). +98.0° to +104°, determined
betamethasone sodium phosphate, C22H28FNa2O8P.
Light absorption (2.4.7). Ratio of the absorbance of the solution
prepared as directed under Assay at the maximum at about Identification
241 nm to that at about 263 nm, 1.70 to 1.90.
Inorganic phosphate. Not more than 0.5 per cent, calculated
as PO4, determined by the following method. Weigh accurately
about 25 mg, dissolve in 10 ml of water, add 4 ml of dilute Mobile phase. A mixture of 60 volumes of butanol, 20 volumes
sulphuric acid, 1 ml of ammonium molybdate solution and of acetic anhydride and 20 volumes of water.
2 ml of methylaminophenol with sulphite solution and allow Test solution. Dilute the eye drops suitably with water to get
to stand for 15 minutes. Add sufficient water to produce a solution containing 0.1 per cent w/v of Betamethasone
25.0 ml, allow to stand for further 15 minutes and measure the Sodium Phosphate.
730 nm (2.4.7). Calculate the content of phosphate from a Reference solution (a). A 0.1 per cent w/v solution of
calibration curve prepared by treating suitable aliquots of a betamethasone sodium phosphate RS in water.
0.00143 per cent w/v solution of potassium dihydrogen Reference solution (b). A mixture of equal volumes of the test
phosphate in a similar manner. solution and reference solution (a).
Free betamethasone and other derivatives. Determine by thin- Reference solution (c). A mixture of equal volumes of reference
layer chromatography, (2.4.17), coating the plate with silica solution (a) and 0.1 per cent w/v of prednisolone sodium
gel GF254. phosphate RS in water.
Mobile phase. Methanol. Apply to the plate 10 µl of each solution. Allow the mobile
Test solution. Dissolve 1.0 g of the substance under phase to rise 10 cm. Dry the plate in air, heat at 110º for 10
examination in 10 ml of methanol. minutes and examine in ultraviolet light at 254 nm. The
182
IP 2007 BETAMETHASONE INJECTION
chromatograms obtained with the test solution, reference (a). Ignore any peak the area of which is less than 0.05 times
solution (a) and reference solution (b) show single principal the area of the principal peak in the chromatogram obtained
spots with similar Rf values. The chromatogram obtained with with reference solution (a).
reference solution (c) shows two principal spots with almost Other tests. Comply with the tests stated under Eye Drops.
identical Rf values.
obtained with the test solution corresponds to the principal Test solution (a). Mix a quantity of the eye drops containing
peak in the chromatogram obtained with the reference solution. 5 mg of Betamethasone Sodium Phosphate with 10 ml of
methanol and dilute to 25.0 ml with water.
C. To a volume containing 0.2 mg of Betamethasone Sodium
Test solution (b). Mix a quantity of the eye drops containing
Phosphate, add slowly 1 ml of sulphuric acid and allow to
5 mg of Betamethasone Sodium Phosphate with 10 ml of a 0.06
stand for 2 minutes. A brownish yellow colour but no red
per cent w/v solution of hydrocortisone (internal standard) in
colour or yellowish green fluorescence is produced.
methanol and dilute to 25.0 ml with water..
Tests Reference solution. Mix 5.0 ml of a 0.1 per cent w/v solution of
betamethasone sodium phosphate RS in water (solution A)
pH (2.4.24).7.0 to 8.5.
and 10 ml of the internal standard solution and dilute to 25.0
Related substances. Determine by liquid chromatography ml with water.
(2.4.14).
Test solution. Dilute the eye drops if necessary to obtain a – a stainless steel column 20 cm x 5 mm, packed with
solution containing 0.1 per cent w/v of Betamethasone Sodium octadecylsilyl silica gel (10 µm) (such as Spherisorb
Phosphate. ODS 1),
Reference solution (a). Dilute 1 volume of the test solution to – mobile phase: a mixture of 55 volumes of citro-
50 volumes with water. phosphate buffer pH 5.0 and 45 volumes of methanol,
– flow rate. 2 ml per minute.
Reference solution (b). A solution containing 0.006 per cent – spectrophotometer set at 241 nm,
w/v each of betamethasone sodium phosphate RS and – a 20 µl loop injector.
betamethasone RS.
Chromatographic system relative standard deviation for replicate injections is not more
– a stainless steel column 20 cm x 4.6 mm, packed with than 2.0 per cent.
octadecylsilyl silica gel (10 µm) (such as Spherisorb Inject test solutions (a), (b) and reference solution.
ODS 1),
– column temperature. 60º, Calculate the content of C22H28FNa2O8P in the eye drops.
– mobile phase: a mixture of 60 volumes of citro- Storage. Store protected from light.
phosphate buffer pH 5.0 and 40 volumes of methanol,
– spectrophotometer set at 241 nm, Betamethasone Injection
Betamethasone Sodium Phosphate Injection
Betamethasone Injection is a sterile solution of Betamethasone
resolution between the peaks due to betamethasone sodium
Sodium Phosphate in Water for Injections.
phosphate and betamethasone is at least 3.5.
Betamethasone Injection contains not less than 92.5 per cent
Inject the test solution and reference solution (a) and record
the chromatogram for three times the retention time of the
betamethasone, C22H29FO5.
principal peak. In the chromatogram obtained with the test
solution the area of any peak corresponding to betamethasone Description. A clear, colourless solution.
is not more than 1.3 times the area of the principal peak in the
Identification
chromatogram obtained with reference solution (a). The area
of any other secondary peak is not more than 1.5 times the A. Determine by thin-layer chromatography (2.4.17), coating
area of the principal peak in the chromatogram obtained with the plate with silica gel G.
reference solution (a). The sum of the areas of all the secondary Mobile phase. A freshly prepared mixture of 30 volumes of
peaks is not greater than 2.5 times the area of the principal 1-butanol, 10 volumes of acetic anhydride and 10 volumes of
peak in the chromatogram obtained with reference solution water.
183
BETAMETHASONE SODIUM PHOSPHATE TABLETS IP 2007
Test solution. Dilute the injection, if necessary, with water so Labelling. The label states the strength in terms of the
that it contains the equivalent of 2 mg of betamethasone per equivalent amount of betamethasone in a suitable dose-volume.
ml.
betamethasone sodium phosphate RS in water. Betamethasone Sodium Phosphate
Reference solution (b). A mixture of equal volumes of the test Tablets
solution and reference solution (a). Betamethasone Sodium Phosphate Tablets contain not less
Reference solution (c). A mixture of equal volumes of the test than 90.0 per cent and not more than 110.0 per cent of the
solution and a 0.25 per cent w/v solution of prednisolone stated amount of betamethasone, C22H29FO5.
sodium phosphate RS in water.
Identification
dry the plate in air until the odour of solvents is no longer A. Determine by thin-layer chromatography (2.4.17), coating
detectable, spray with ethanolic sulphuric acid (20 per cent), the plate with silica gel G.
heat at 120° for 10 minutes, allow to cool, and examine in Mobile phase. A freshly prepared mixture of 30 volumes of
ultraviolet light at 365 nm. The principal spot in the 1-butanol, 10 volumes of acetic anhydride and 10 volumes of
chromatogram obtained with the test solution corresponds to water.
that in the chromatogram obtained with reference solution (a).
Test solution. Dissolve a quantity of the powdered tablets
The principal spot in the chromatogram obtained with
containing 2 mg of betamethasone in 25 ml of water, add 2.5 g
reference solution (b) appears as a single, compact spot and
of sodium chloride and 1 ml of hydrochloric acid, extract
the chromatogram obtained with reference solution (c) shows
with 25 ml of chloroform and discard the chloroform layer.
two closely running spots. Secondary spots due to excipients
Extract with 2.5 ml of tributyl phosphate and discard the
may also be seen in the chromatograms obtained with the test
aqueous layer.
solution and reference solutions (b) and (c).
Reference solution (a). Prepare in the same manner as the
B. To a volume containing 4 mg of betamethasone, add 1 ml of test solution but using 2.5 mg of betamethasone sodium
water and sufficient ethanol to produce 40 ml. To 2 ml of this phosphate RS instead of the substance under examination.
solution in a stoppered tube add 10 ml of phenylhydrazine
solution, mix, warm in a water-bath at 60° for 20 minutes and Reference solution (b). A mixture of equal volumes of the test
cool immediately; absorbance of the resulting solution at the solution and reference solution (a).
maximum at about 450 nm, not more than 0.1 (2.4.7). Reference solution (c). A mixture of equal volumes of the test
solution and a solution prepared in the same manner as the
Tests test solution but using 2.5 mg of prednisolone sodium
pH (2.4.24). 7.5 to 9.0 phosphate RS instead of the substance under examination.
dry the plate in air until the odour of solvents is no longer
detectable, spray with ethanolic sulphuric acid (20 per cent),
Assay. Measure accurately a volume containing about 20 mg heat at 120° for 10 minutes, allow to cool, and examine in
of betamethasone and add sufficient water to produce 50.0 ultraviolet light at 365 nm. The principal spot in the
ml. To 5.0 ml add 20 ml of water and 2 ml of 0.1 M hydrochloric chromatogram obtained with the test solution corresponds to
acid and shake with two quantities, each of 25 ml, of ether. that in the chromatogram obtained with reference solution (a).
Wash the ethereal solutions separately with 2, 1 and 1 ml of The principal spot in the chromatogram obtained with
water, add the washings to the acid solution and discard the reference solution (b) appears as a single, compact spot and
ether solutions. To the combined acid solution and the the chromatogram obtained with reference solution (c) shows
washings add 2 ml of 0.1 M sodium hydroxide and sufficient two closely running spots. Secondary spots due to excipients
water to produce 200.0 ml. Measure the absorbance of the may also be seen in the chromatograms obtained with the test
resulting solution at the maximum at about 241 nm (2.4.7), solution and reference solutions (b) and (c).
using as the blank a solution prepared in a similar manner but
B. Mix a quantity of the powdered tablets containing 0.4 mg of
omitting the substance under examination. Calculate the
betamethasone with 1 ml of sulphuric acid and allow to stand
content of C22H29FO5 taking 391 as the specific absorbance at
for 5 minutes; a pale yellow colour is produced.
241 nm.
Tests
exceeding 30°. Disintegration (2.5.1). Maximum time, 5 minutes.
184
IP 2007 BETAMETHASONE VALERATE
Uniformity of content. Comply with the test stated under Betamethasone Valerate contains not less than 96.0 per cent
Tablets. Determine by liquid chromatography (2.4.14). and not more than 102.0 per cent of C27H37FO6, calculated on
Test solution. Powder one tablet and dissolve as completely the dried basis.
as possible in 5 ml of water and add 5 ml of a 0.006 per cent Description. A white to creamy-white powder.
w/v solution of hydrocortisone (internal standard) in
methanol. Identification
Reference solution. A mixture of equal volumes of a 0.0065 per
cent w/v solution of betamethasone sodium phosphate RS in
Compare the spectrum with that obtained with betamethasone
water and a 0.006 per cent w/v solution of hydrocortisone in
valerate RS or with the reference spectrum of betamethasone
methanol.
valerate.
octadecylsilyl silica gel (5 µm) (such as Spherisorb ODS),
– mobile phase: a mixture of 55 volumes of citro- Solvent mixture. A mixture of 90 volumes of acetone and
phosphate buffer pH 5.0 and 45 volumes of methanol, 10 volumes of formamide.
10 volumes of chloroform.
Calculate the content of C22H29FO5 in the tablet. Test solution. Dissolve 25 mg of the substance under
Assay. Determine by liquid chromatography (2.4.14) Reference solution (a). Dissolve 25 mg of betamethasone RS
in 10 ml of the solvent mixture.
Test solution. Weigh and powder 20 tablets. To a quantity of
the powder containing 1.25 mg of betamethasone add 25.0 ml Reference solution (b). Mix equal volumes of the test solution
of a 0.006 per cent w/v solution of hydrocortisone (internal and reference solution (a).
standard) in methanol and dilute to 50.0 ml with water. Place the dry plate in a tank containing a shallow layer of the
Reference solution. A mixture of equal volumes of a 0.0065 per solvent mixture, allow the solvent mixture to ascend to the
cent w/v solution of betamethasone sodium phosphate RS in top, remove the plate from the tank and allow the solvent to
water and a 0.006 per cent w/v solution of hydrocortisone in evaporate. Use within 2 hours, with the flow of the mobile
methanol. phase in the direction in which the aforementioned treatment
Carry out the chromatographic procedure described under was done.
Uniformity of content. Calculate the content of C22H29FO5 in Apply to the plate 1 µl of each solution. Allow the mobile
the tablets. phase to rise 12 cm. Dry the plate in a current of warm air, allow
Storage. Store protected from light. the solvent to evaporate, heat at 120° for 15 minutes and spray
Labelling. The label states the strength in terms of the the hot plate with ethanolic sulphuric acid (20 per cent v/v).
equivalent amount of betamethasone. Heat at 120° for a further 10 minutes, allow to cool and examine
in daylight and in ultraviolet light at 365 nm. The principal
spot in the chromatogram obtained with the test solution
Betamethasone Valerate corresponds to that in the chromatogram obtained with
reference solution (a). The principal spot in the chromatogram
O obtained with reference solution (b) appears as a single,
OH
H3 C compact spot.
HO OCO(CH2)3CH3
C. In the Assay, the retention time of the principal peak in the
H3C H CH3 chromatogram obtained with the test solution corresponds to
that of the peak due to betamethasone valerate RS in the
F H
O
D. Heat 50 mg with 2 ml of 0.5 M ethanolic potassium
C27H37FO6 Mol. Wt. 476.6 hydroxide in a water-bath for 5 minutes. Cool, add 2 ml of
Betamethasone Valerate is a 9α−fluoro-11β,17α,21-trihydroxy- sulphuric acid (50 per cent v/v) and boil gently for 1 minute;
16β-methyl-pregna-1,4-diene-3,20-dione-17-valerate. the odour of ethyl valerate is perceptible.
185
BETAMETHASONE VALERATE OINTMENT IP 2007
Tests – mobile phase: a mixture of 30 volumes of acetonitrile

and 20 volumes of water,
Specific optical rotation (2.4.22). +75.0° to +82.0°, determined – flow rate. 1.2 ml per minute,
in a 1.0 per cent w/v solution in dioxan. – spectrophotometer set at 254 nm,
Light absorption (2.4.7). Absorbance of a 0.002 per cent w/v – a 10 µl loop injector.
solution in ethanol at the maximum at about 240 nm, 0.63 to Inject reference solution (a). The relative retention times are
0.67. about 1.7 for beclomethasone dipropionate and 1.0 for
Related substances. Determine by liquid chromatography betamethasone valerate,The resolution between
(2.4.14). betamethasone valerate and beclomethasone dipropionate is
Test solution. Weigh accurately 4 mg of the substance under not less than 4.5 and the relative standard deviation for
examination add 10 ml of the mobile phase and shake well to replicate injections is not more than 2.0 per cent.
dissolve. Inject alternately the test solution and the reference solution.
Chromatographic system Calculate the percentage content of C27H37FO6.
– a stainless steel column 15 cm × 4.6 mm packed with Storage. Store protected from light.
octadecylsilyl silica gel (3 to 10 µm),
45 volumes of water and 0.1 volume of glacial acetic Betamethasone Valerate Ointment
acid,
– flow rate. 1 ml per minute, Betamethasone Valerate Ointment contains Betamethasone
– spectrophotometer set at 254 nm, Valerate in a suitable ointment base.
– a 10 µl loop injector. Betamethasone Valerate Ointment contains not less than 90.0
Inject the test solution. The resolution between betamethasone per cent and not more than 110.0 per cent of the stated amount
valerate and any impurity is not less than 1.5 and the column of betamethasone, C22H29FO5.
efficiency is not less than 9000 theoretical plates.
Identification
Inject the test solution. Measure all the peak responses.
Calculate the content of each impurity as a percentage of the A. Determine by thin-layer chromatography (2.4.17), coating
sum of all the peak responses (1.0 per cent). Not more than the plate with silica gel G.
2.0 per cent of total impurities is found. Mobile phase. A mixture of 20 volumes of chloroform, 2
Sulphated ash (2.3.18). Not more than 0.1 per cent. volumes of acetone and 1 volume of ethanol.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Test solution. Heat a quantity of the ointment containing 1 mg
on 1.0 g by drying in an oven at 105°. of betamethasone with 10 ml of methanol on a water-bath
until it boils, shake vigorously, cool in ice for 30 minutes, filter,
Assay. Determine by liquid chromatography (2.4.14). evaporate the filtrate to dryness in a current of nitrogen with
Test solution. Weigh accurately about 60 mg of the substance gentle heating and dissolve the residue in 0.5 ml of chloroform.
under examination, dissolve in a 0.1 percent v/v solution of Reference solution. A 0.24 per cent w/v solution of
glacial acetic acid in methanol and dilute to 100.0 ml with the betamethasone valerate RS in chloroform.
same solvent. To 5.0 ml of this solution add 10.0 ml of reference
solution (b) and mix. Apply to the plate 10 µl of each solution. After development,
dry the plate in air, heat at 105° for 5 minutes and spray while
Reference solution (a). Weigh accurately a suitable quantity hot with alkaline tetrazolium blue solution. The principal
of betamethasone valerate RS and dissolve in a 0.1 per cent spot in the chromatogram obtained with the test solution
v/v solution of glacial acetic acid in methanol to obtain a corresponds to that in the chromatogram obtained with the
solution containing a known concentration of about 0.6 mg reference solution.
per ml. To 5.0 ml of this solution add 10.0 ml of reference
solution (b) and mix. B. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak due
Reference solution (b). A 0.04 per cent w/v solution of to betamethasone valerate RS in the chromatogram obtained
betamethasone dipropionate RS in a 0.1 per cent v/v solution with the reference solution.
of glacial acetic acid in methanol.
Tests
– a stainless steel column 30 cm × 4.0 mm, packed with Microbial contamination (2.2.9). 1.0 g is free from
octadecylsilyl silica gel (3 to 10 µm), Staphylococcus aureus and Pseudomonas aeruginosa.
186
IP 2007 BIPERIDEN HYDROCHLORIDE
Other tests. Complies with the tests stated under Ointments. Biperiden Hydrochloride contains not less than 99.0 per cent
Assay. Determine by liquid chromatography (2.4.14). and not more than 101.0 per cent of C21H29NO,HCI, calculated
on the dried basis.
Test solution. Heat a quantity of the accurately weighed
ointment containing 2.5 mg of betamethasone with 10.0 ml of
0.04 per cent w/v solution of beclomethasone dipropionate
RS (internal standard) in methanol containing 0.1 per cent
Identification
v/v of glacial acetic acid and 5.0 ml of methanol containing Test A may be omitted if tests B, C and D are carried out. Tests
0.1 per cent v/v of glacial acetic acid on a water-bath until it B and C may be omitted if tests A and D are carried out.
boils, shake vigorously, cool in ice for 30 minutes, centrifuge
and decant the supernatant solution into a stoppered flask. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with biperiden
Reference solution. Mix 5 ml of a 0.06 per cent w/v solution of hydrochloride RS.
betamethasone valerate RS in methanol containing 0.1 per-
cent v/v of glacial acetic acid and 10.0 ml of a 0.04 per cent B. Determine by thin-layer chromatography (2.4.17), coating
w/v solution of beclomethasone dipropionate RS in methanol the plate with silica gel GF254.
containing 0.1 per cent v/v of glacial acetic acid. Mobile phase. A mixture of 100 volumes of toluene, 5 volumes
Chromatographic system of diethylamine and 5 volumes of methanol.
– a stainless steel column 30 cm × 4.0 mm, packed with Test solution. Dissolve 0.5 g of the substance under
octadecylsilyl silica gel (5 µm), examination in 100 ml of methanol.
and 40 volumes of water, Reference solution (a). A 0.5 per cent w/v solution of
– flow rate. 1.2 ml per minute, biperiden hydrochloride RS in methanol.
– spectrophotometer set at 240 nm, Reference solution (b). Dissolve 5 mg of (SR)-1-[(1RS, 2RS,
– a 20 µl loop injector. 4RS)-bicyclo [2.2.1] hept-5-en-2-yl]-1-phenyl-3-(piperidin-1-
Inject the reference solution. The relative retention times yl]-propan-1-ol (endo form) in reference solution (a) and dilute
should be about 1.7 for beclomethasone dipropionate and 1.0 to 2 ml with the same solution.
for betamethasone valerate. Apply to the plate 5 µl of each solution. After development,
Inject alternately the test solution and the reference solution. dry the plate in air and examine in ultraviolet light at 254 nm.
The principal spot in the chromatogram obtained with the test
Calculate the content of C22H29FO5 in the ointment. solution corresponds to that in the chromatogram obtained
Storage. Store protected from light. Avoid exposure to with reference solution (a). Spray with dilute iodobismuthate
excessive heat. solution and examine in daylight. The principal spot in the
equivalent amount of betamethasone.
The test is not valid unless the chromatogram obtained with
reference solution (b) shows two clearly separated spots.
C. To about 20 mg add 5 ml of phosphoric acid; a green colour
Biperiden Hydrochloride develops.
D. Gives reaction A of chlorides (2.3.1).
Tests
N
,HCl
HO Appearance of solution. A 0.2 per cent solution in carbon
dioxide-free water is not more opalescent than opalescence
standard OS2 (2.4.1), and is colourless (2.4.1).
C21H29NO,HCI Mol. wt. 347.9 Related substances. Determine by gas chromatography
Biperiden Hydrochloride is (RS)-1-[(RS,2RS,4RS)-bicyclo- (2.4.13).
[2.2.1]hept-5en-2-yl]-1-phenyl-3-(piperidin-1-yl)propan-1-ol Test solution. Dissolve 1.0 g of the substance under
hydrochloride. examination in 100 ml of methanol.
187
BIPERIDEN TABLETS IP 2007
Reference solution (a). Dilute 1.0 ml of the test solution to to effect solution and cool. Add 10 ml of mercuric acetate
100 ml with methanol and mix. Dilute 10 ml of the resulting solution and titrate with 0.1 M perchloric acid, using 0.1ml
solution to 100 ml with methanol. of crystal violet solution as indicator. Carry out a blank
titration.
Reference solution (b). To 1.0 ml of the test solution add 10 ml
of methanol and 10 mg of (SR)-1-[(1RS, 2RS, 4RS)- 1 ml of 0.1 M perchloric acid is equivalent to 0.03479 g of
bicyclo[2.2.1]hept-5-en-2-yl]-1-phenyl-3-(piperidin-1-yl) C21H29NO, HCl.
propan-l-ol (endo form)and sufficient methanol to produce Storage. Store protected from light.
100 ml.
– a fused-silica capillary column, 50 m × 0.25 mm coated
with poly (vinyl-phenylmethyl siloxane with thickness Biperiden Tablets
of 0.25 µm,
– flame ionisation detector,
Biperiden Hydrochloride Tablets
– temperature: Biperiden Tablets contain not less than 92.5 per cent and not
– column. 200° for 5 minutes, then raised at the rate of more than 107.5 per cent of the stated amount of biperiden
2° per minute to 270°, hydrochloride, C21H29NO, HCl.
– inlet port at 250° and detector at 300°,
– flow rate. 0.4 ml per minute of nitrogen (the carrier gas) Identification
and a split ratio of 1:250. Determine by thin-layer chromatography (2.4.17), coating the
Inject 2 µl of each solution. When using a recorder, adjust the plate with silica gel G.
sensitivity of the system so that the heights of the two principal
Mobile phase. A mixture of 100 volumes of methanol and
peaks in the chromatogram obtained with reference solution
1.5 volumes of strong ammonia solution.
(b) are not less than 50 per cent of the full scale of the recorder.
The test is not valid unless, in the chromatogram obtained Test solution. Shake a quantity of the powdered tablets
with reference solution (b), the resolution between the first containing about 10 mg of Biperiden Hydrochloride with 5 ml
peak due to biperiden and the second peak due to (SR)-1- of water and disperse the powder with the aid of ultra sound
[(1RS, 2RS,4RS)]-bicyclo [2.2.1] hept-5-en-2-yl]-phenyl-3- for a few minutes. Add 5 ml of methanol and mix again for
(piperidin-1-yl)propa-1-ol (endo form) is at least 2.5; the 15 minutes. Filter the solution into a separator, add 2 ml of 1 M
principal peak in the chromatogram obtained with reference sodium hydroxide and 10 ml of chloroform and shake for
solution (a) has a signal-to-noise ratio of at least 6. For peaks 3 minutes. Filter the chloroform layer into a stoppered flask
with a retention time of 0.95 to 1.05 relative to biperiden, the and use the filtrate.
Reference solution. Prepare in a similar manner using 10 mg of
than 0.5 per cent of the area of the principal peak and the sum
biperiden hydrochloride RS in place of the substance under
of the areas of any such peaks is not greater than 1.0 per cent
examination.
of the area of the principal peak. For peaks with relative
retention times outside the above-mentioned range, the area Apply to the plate 20 µl of each solution. After development,
of any peak is not greater than 0.1 per cent of the area of the dry the plate in air and expose it to iodine vapours till spots
principal peak and the sum of the areas of such peaks is not appear. The principal spot in the chromatogram obtained with
greater than 0.5 per cent of the area of the principal peak. the test solution corresponds to that in the chromatogram
Disregard any peak with an area less than 0.05 per cent of the obtained with the reference solution.
the test solution. Tests
Heavy metals (2.3.13). 1.0 g complies with the limit test for Dissolution (2.5.2).
heavy metals, Method D (20 ppm).
Apparatus. No 1
Sulphated ash (2.3.18). Not more than 0.1 per cent. Medium. 500 ml of 0.1 M hydrochloric acid.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Speed and time. 50 rpm and 45 minutes.
on 1.0 g by drying in an oven at 105° for 2 hours. Use one tablet in the vessel for each test.
Assay. Weigh accurately about 0.5 g, dissolve in 80 ml of Withdraw 75 ml of the solution and filter through a membrane
anhydrous glacial acetic acid, warming slightly, if necessary filter disc with an average pore diameter not greater than
188
IP 2007 BISACODYL
1.0 µm, rejecting the first few ml of the filtrate. Transfer 50.0 ml Bisacodyl
of the clear filtrate into a suitable container, adjust the pH to
5.3 with 0.1 M sodium hydroxide. Transfer this solution to a
100-ml volumetric flask and dilute with water to volume and
mix.
Prepare a reference solution by weighing accurately about N
80 mg of biperiden hydrochloride RS in sufficient methanol
to produce 100.0 ml. Dilute 5.0 ml of this solution to 500.0 ml O O
with 0.1 M hydrochloric acid and mix. Transfer 25.0 ml of the
resulting solution into a suitable container and adjust the pH H3C O O CH3
to 5.3 with 0.1 M sodium hydroxide and dilute to 100.0 ml with
water (2 µg per ml).
Prepare a blank solution by treating 50 ml of water in place of C22H19NO4 Mol. Wt. 361.4
the clear filtrate in the same manner as described for the test
solution beginning at the words “adjust the pH to 5.3…… Bisacodyl is bis(4-acetoxyphenyl)-2-pyridylmethane.
Transfer 20.0 ml of the solutions into individual separators, Bisacodyl contains not less than 98.0 per cent and not more
each containing 10.0 ml of phosphate-buffered bromocresol than 101.0 per cent of C22H19NO4, calculated on the dried basis.
purple solution. Add 40.0 ml of chloroform to each and shake Description. A white or almost white, crystalline powder;
for 10 minutes. After the layers have separated, filter each odourless.
chloroform extract through a filter paper into separate, glass-
stoppered flasks, discarding the first 10 ml of each filtrate.
Identification
Measure the absorbances of the solutions at the maximum at
about 408 nm against the blank solution (2.4.7). Calculate the A. Determine by infrared absorption spectrophotometry (2.4.6).
content of C21H29NO,HCl in the medium from the absorbance Compare the spectrum with that obtained with bisacodyl RS
obtained from the reference solution. or with the reference spectrum of bisacodyl.
D: Not less than 75 per cent of the stated amount of C21H29NO, B. When examined in the range 230 nm to 360 nm, a 0.001 per
HCl. cent w/v solution in 0.1 M potassium hydroxide in methanol
shows an absorption maximum only at about 248 nm;
Other tests. Comply with the tests stated under Tablets. absorbance at about 248 nm, about 0.65 (2.4.7).
quantity of the powder containing about 2 mg of Biperiden Tests
Hydrochloride and transfer to a 50-ml volumetric flask, add
12.5 ml of water and heat on a steam-bath for 15 minutes. Acidity or alkalinity. Shake 1.0 g with 20 ml of carbon dioxide-
Cool, dilute with methanol to volume and mix. Transfer 5.0 ml free water, boil, cool and filter. Add 0.2 ml of 0.01 M sodium
of the resulting solution to a separator, add 10.0 ml of hydroxide and 0.1 ml of methyl red solution. The resulting
phosphate-buffered bromocresol purple solution, extract with solution is yellow and not more than 0.4 ml of 0.01 M
two quantities, each of 20 ml, of chloroform and allow to hydrochloric acid is required to change the colour of the
separate. Filter the chloroform extracts into a 50-ml volumetric solution to red.
flask through filter paper and make to volume. Measure the
408 nm (2.4.7), using a reagent blank of a mixture of 3 volumes Loss on drying (2.4.19). Not more than 0.5 per cent, determined
of methanol and 1 volume of water and preparing the solution on 0.5 g by drying in an oven at 105°.
in a similar manner as that of the test solution omitting the
substance under examination. Calculate the content of
C21H29NO, HCl from the absorbance obtained by repeating
the operation using a solution prepared by adding 5.0 ml of a
Carry out a blank titration.
0.08 per cent w/v solution of biperiden hydrochloride RS in
methanol to 25 ml of water, diluting to 100.0 ml with methanol 1 ml of 0.1 M perchloric acid is equivalent to 0.03614 g of
and treating in the same manner as the test solution. C22H19NO4.
Storage. Store protected from light. Storage. Store protected from light.
189
BISACODYL SUPPOSITORIES IP 2007
Bisacodyl Suppositories w/v solution of sulphuric acid (solution A). To 2 ml of the

solution add one drop of potassium mercuri-iodide solution;
Bisacodyl Suppositories contain Bisacodyl in a suitable a white precipitate is produced.
suppository basis.
B. To 2 ml of solution A add sulphuric acid; a reddish-violet
Bisacodyl Suppositories contain not less than 90.0 per cent colour is produced.
C. Boil 2 ml of solution A with a few drops of nitric acid; a
bisacodyl, C22H19NO4.
yellow colour is produced. Cool and add 5 M sodium
Identification hydroxide; the colour becomes yellowish-brown.
A. Dissolve a quantity of the suppositories containing 0.15 g Tests

of Bisacodyl as completely as possible in 150 ml of light
petroleum (40° to 60°), filter, wash the residue with light Uniformity of content. Comply with the test stated under
petroleum (40° to 60°) until free from fatty material and dry at Tablets.
about 100°. Wash with a very small quantity of warm Powder one tablet, shake with 70 ml of chloroform for
chloroform and dissolve the residue in 10 ml of a 1 per cent 30 minutes and dilute with sufficient chloroform to produce
w/v solution of sulphuric acid (solution A). To 2 ml of the 100.0 ml. Mix well, filter and discard the first few ml of the
solution add one drop of potassium mercuri-iodide solution; filtrate. Measure the absorbance of the filtrate at the maximum
a white precipitate is produced. at about 264 nm (2.4.7). Calculate the content of C22H19NO4
taking 148 as the specific absorbance at 264 nm.
B. To 2 ml of the solution A add sulphuric acid; a reddish
violet colour is produced. Other tests. Comply with the tests stated under Tablets.
C. Boil 2 ml of the solution A with a few drops of nitric acid; a In the test for Disintegration, use a 1.5 per cent w/v solution of
yellow colour is produced. Cool and add 5 M sodium sodium bicarbonate in place of mixed phosphate buffer pH 6.8.
hydroxide; the colour becomes yellowish brown. Assay. Weigh and powder 20 tablets. Weigh accurately a
quantity of the powder containing about 40 mg of Bisacodyl,
Tests
shake with 70 ml of chloroform for 30 minutes, dilute to
Other tests. Comply with the tests stated under Suppositories. 100.0 ml with chloroform, mix, filter and dilute 10.0 ml of the
filtrate to 100.0 ml with chloroform. Measure the absorbance
Assay. Weigh accurately a quantity of the suppositories
of the resulting solution at the maximum at about 264 nm (2.4.7).
containing about 50 mg of Bisacodyl, add 80 ml of anhydrous
Calculate the content of C22H19NO4 taking 148 as the specific
glacial acetic acid previously neutralised with 0.02 M
perchloric acid to 1-naphtholbenzein solution and warm
gently until solution is complete. Immediately titrate with
0.02 M perchloric acid, determining the end-point
potentiometrically (2.4.25). Carry out a blank titration. Bismuth Subcarbonate
1 ml of 0.02 M perchloric acid is equivalent to 0.007228 g of Bismuth Carbonate
C22H19NO4.
Bismuth Subcarbonate contains not less than 80.0 per cent
Storage. Store protected from light at a temperature not and not more than 82.5 per cent of Bi, calculated on the dried
exceeding 30°. basis.
Description. A white or almost white powder; odourless.
Bisacodyl Tablets Identification

Bisacodyl Tablets contain not less than 95.0 per cent and not A. Gives the reactions of bismuth salts (2.3.1).
more than 105.0 per cent of the stated amount of bisacodyl, B. Gives reaction A of carbonates (2.3.1).
C22H19NO4. The tablets are rendered gastro-resistant by enteric
coating or by other means. Tests
Identification Appearance of solution. Shake 5.0 g with 10 ml of water, add
20 ml of nitric acid. Heat to dissolve, cool and dilute to 100 ml
A. Extract a quantity of the powdered tablets containing with water (solution A). Solution A is not more opalescent
50 mg of Bisacodyl with chloroform, filter, evaporate the filtrate than opalescence standard OS2 (2.4.1), and is colourless
to dryness and dissolve the residue in 10 ml of a 1 per cent (2.4.1).
190
IP 2007 BLEOMYCIN SULPHATE
Alkalis and alkaline-earth metals. Not more than 1.0 per cent, acid and heat to 70°, maintaining the solution at this
determined by the following method. To 1.0 g add 10 ml of temperature until the solution becomes completely clear. Add
water and 10 ml of 5 M acetic acid, boil for 2 minutes, cool, about 50 mg of xylenol orange mixture and titrate with 0.1 M
filter and wash the residue with 20 ml of water. To the combined disodium edetate until the colour changes from pinkish-violet
filtrate and washings add 2 ml of 2 M hydrochloric acid and to lemon yellow.
20 ml of water. Boil, pass hydrogen sulphide through the 1 ml of 0.1 M disodium edetate is equivalent to 0.02090 g of Bi.
boiling solution until no further precipitate is produced, filter
and wash the residue with water. Evaporate the combined Storage. Store protected from light.
filtrate and washings to dryness on a water-bath and add
0.5 ml of sulphuric acid, ignite gently and allow to cool.
Arsenic (2.3.10). To 0.5 g in a distillation flask add 5 ml of Bleomycin Sulphate
water and 7 ml of sulphuric acid, cool and add 5 g of hydrazine
reducing mixture and 10 ml of hydrochloric acid. Connect
the flask to an air-condenser, heat gradually to boiling during H2N O
H NH2 R
15 to 30 minutes and continue heating at such a rate that the N NH2
O
distillation proceeds steadily and until the volume in the flask
O N
is reduced by half, or until 5 minutes after the condenser has N N CH3 H O S
become full of steam. Discontinue distillation before fumes of O HO N ,H2SO4
H2N O NH N
sulphur trioxide are evolved. Collect the distillate in a tube H3C HN O
S
N CH3 HO CH3
containing 15 ml of water cooled in ice. Wash the condenser O H
N
with water and dilute the combined distillate and washings to HO OH
Bleomycin R = Terminal amine
25 ml with water. The resulting solution complies with the O N
CH3
H
limit test for arsenic (5 ppm). Use 2.5 ml of arsenic standard OH O OH S
A2 N CH3
solution (1 ppm As) diluted to 25 ml with water to prepare the OH H
OH O
standard. B2
O N
NH2
H NH
Copper. To 5 ml of solution A add 2 ml of 10 M ammonia,
HN CNH2
dilute to 50 ml with water and filter. To 10 ml of the filtrate add
1 ml of a 0.1 per cent w/v solution of sodium diethyldithio-
carbamate. Any colour produced is not more intense than C55H84N17O21S3,H2SO4 Mol. Wt. 1516.6
that produced by treating at the same time and in the same (Bleomycin A2 Sulphate)
manner a solution containing 0.25 ml of copper standard C55H84N20O21S2,H2SO4 Mol. Wt. 1523.6
solution (10 ppm Cu) diluted to 10 ml with water (50 ppm). (Bleomycin B2 Sulphate)
Lead. To 10 ml of solution A add 10 ml of 1 M sulphuric acid;
Bleomycin Sulphate is the sulphate salt of bleomycin, a mixture
the solution does not become cloudy.
of basic cytotoxic glycopeptides produced by the growth of
Silver. To 2.0 g add 1 ml of water and 4 ml of nitric acid. Heat Streptomyces verticillus or produced by other means. Its main
gently to dissolve and dilute to 11 ml with water. Cool, add components are bleomycin A2 and bleomycin B2. Bleomycin
2 ml of 1 M hydrochloric acid and allow to stand for 5 minutes A2 sulphate is N1-[3-(dimethylsulphonio)propyl]bleomycina-
protected from light. Any opalescence produced is not more mide hydrogen sulphate and Bleomycin B 2 is N 1 -
intense than that obtained by treating at the same time and in (guanidinobutyl)bleomycinamide sulphate.
the same manner a mixture of 10 ml of silver standard solution Bleomycin Sulphate contains not less than 1.5 and not more
(5 ppm Ag), 2 ml of 1 M hydrochloric acid and 1 ml of nitric than 2.0 Units of bleomycin per mg and the content of
acid (25 ppm). bleomycins is: bleomycin A2, between 55 per cent and 70 per
Chlorides (2.3.12). To 10 ml of solution A add 4 ml of nitric cent;bleomycin B2, between 25 per cent and 32 per cent; sum
acid and 20 ml of water; the resulting solution complies with of bleomycin A2 and bleomycin B2, not less than 85 per cent;
the limit test for chlorides (500 ppm). demethylbleomycin A2, not more than 5.5 per cent; other
related substances, not more than 9.5 per cent.
on 1.0 g by drying in an oven at 105°. Description. A white or cream-coloured, amorphous powder.
Assay. Weigh accurately about 0.5 g, dissolve in 3 ml of nitric CAUTION — Bleomycin Sulphate must be handled with care,
acid and dilute to 250 ml with water. Add strong ammonia avoiding contact with the skin and inhalation of airborne
solution until cloudiness is first observed, add 0.5 ml of nitric particles.
191
BLEOMYCIN INJECTION IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). After the final conditions are reached (about 60 minutes) allow
Compare the spectrum with that obtained with bleomycin the chromatography to proceed with the final gradient mixture
sulphate RS. for an additional 20 minutes or until demethylbleomycin A2 is
eluted.
B. Gives the reactions of sulphates (2.3.1).
Inject the test solution and proceed with gradient elution,
Tests pumping the mobile phase mixture under the conditions
mentioned above for about 80 minutes or until the
pH (2.4.24). 4.5 to 6.0, determined in a solution containing 10 demethylbleomycin A2 is eluted. The usual order of elution is
Units per ml. bleomycinic acid, bleomycin A2 (first principal peak), bleomycin
Copper. Not more than 0.02 per cent determined by Method A A5, bleomycin B2 (second principal peak), bleomycin B4 and
or by Method B. demethylbleomycin A2 (retention time relative to bleomycin
A2, between 1.5 and 2.5).
A. Weigh accurately about 50 mg, transfer to a 60-ml separator
and dissolve in 10.0 ml of 0.1 M hydrochloric acid. Add 10 ml Measure the peak responses of all peaks. Calculate the
of a 0.01 per cent w/v solution of zinc bis (diphenyl percentage contents of each bleomycin component by
dithiocarbamate) in carbon tetrachloride and shake comparing the ratios of the individual areas of the peaks with
vigorously for 1 minute. Allow the layers to separate, filter the that of the total area of all bleomycins.
lower layer through 1 g of anhydrous sodium sulphate. Treat Loss on drying (2.4.19). Not more than 6.0 per cent, determined
similarly 10.0 ml of copper standard solution (10 ppm Cu) on 0.5 g by drying in an oven over phosphorous pentoxide at
and measure the absorbances (2.4.7) of the two solutions at 60° at a pressure not exceeding 0.25 kPa for 3 hours.
the maximum at about 435 nm, using carbon tetrachloride as
the blank. Assay. Determine by the microbiological assay of antibiotics,
Method A or B (2.2.10), and express the result in Units per mg.
B. Determine by atomic absorption spectrophotometry (2.4.2)
measuring at 324.7 nm using an air-acetylene flame and a Bleomycin Sulphate intended for use in the manufacture of
solution prepared in the following manner: Dissolve about 50 parenteral preparations without a further appropriate
mg of the substance under examination in water and dilute to procedure for the removal of bacterial endotoxins complies
10 .0 ml with the same solvent. Use copper solution AAS with the following additional requirement.
suitably diluted with water, for preparing the reference Bacterial endotoxins (2.2.3). Not more than 10.0 Endotoxin
solutions. Units per unit of bleomycin.
Content of bleomycins. Determine by liquid chromatography Bleomycin Sulphate intended for use in the manufacture of
(2.4.14). parenteral preparations without a further appropriate
Test solution. Dissolve the substance under examination in sterilisation procedure complies with the following
freshly boiled and cooled water so as to give a solution additional requirement.
containing about 2.5 Units per ml. This solution should be Sterility (2.2.11). Complies with the test for sterility.
stored at 2° to 8° until just before use.
Storage. If the material is sterile, it should be stored in sterile,
Chromatographic system tamper-evident containers and sealed so as to exclude micro-
– a stainless steel column 25 cm × 4.6 mm, packed with organisms.
octadecylsilyl silica gel (5 µm) (such as Nucleosil C18),
Labelling. The label states (1) the strength with respect to
– mobile phase: Transfer 0.96 g of sodium
Bleomycin Sulphate as the number of bleomycin Units per
1-pentanesulphonate to a 1000-ml volumetric flask, add
mg; (2) whether or not the contents are intended for use in the
5.0 ml of glacial acetic acid and 900 volumes of water.
manufacture of parenteral preparations.
Mix and adjust the pH to 4.3 with strong ammonia
solution (1.86 g of disodium edetate may be included if
needed for satisfactory chromatography). Adjust the
volume with water, mix well, filter and degas before use. Bleomycin Injection
Use a linear gradient of 10 per cent to 40 per cent
methanol, which also is filtered and degassed before Bleomycin sulphate injection
use, mixed with this solution, Beomycin injection is a sterile freeze dried material consisting
– flow rate. 1.5 ml per minute, of Bleomycin sulphate with or without excipients. It is filled in
– spectrophotometer set at 254 nm, a sealed container.
192
IP 2007 BLEOMYCIN INJECTION
The injection is constituted by dissolving the contents of the Content of bleomycins. Determine by liquid chromatography
sealed container in the requisite amount of the liquid stated (2.4.14).
on the label before use. Test solution. Weigh accurately a suitable quantity dissolve
The constituted solution complies with the tests for Clarity in freshly boiled and cooled water and dilute to obtain a
of solution and Particulate matter stated under Parenteral solution containing about 2.5 Units per ml. This solution
Preparations (Injections). should be stored at 2° to 8° until just before use.
Storage. The constituted solution should be used immediately Chromatographic system
after preparation but, in any case, within the period – a stainless steel column 25 cm × 4.6 mm, packed with
recommended by the manufacturer. octadecylsilyl silica gel (5 µm) (such as Nucleosil 7C18),
– mobile phase: Transfer 0.96 g of sodium
Bleomycin injection contains not less than 90.0 per cent and
1-pentanesulphonate to a 1000-ml volumetric flask, add
not more than 120.0 per cent of the stated amount of bleomycin
5.0 ml of glacial acetic acid and 900 volumes of water.
and the content of bleomycins is: bleomycin A2, between
Mix and adjust the pH to 4.3 with strong ammonia
55 per cent and 70 per cent;bleomycin B2, between 25 and
solution (1.86 g of disodium edetate may be included if
32 per cent; sum of bleomycin A2 and bleomycin B2, not less
needed for satisfactory chromatography). Adjust the
than 85 per cent; demethylbleomycin A2, not more than 5.5 per
volume with water, mix well, filter and degas before use.
cent; other related substances, not more than 9.5 per cent.
Use a linear gradient of 10 per cent to 40 per cent
The contents of the sealed container comply with the tests methanol, which also is filtered and degassed before
stated under Parenteral Preparations (Powders for use, mixed with this solution,
Injection) and with the following requirements. – flow rate. 1.5 ml per minute,
After the final conditions are reached (about 60 minutes) allow
A. Determine by infra-red absorption spectrophotometry
the chromatography to proceed with the final gradient mixture
(2.4.6). Compare the spectrum with that obtained with
for an additional 20 minutes or until demethylbleomycin A2 is
bleomycin sulphate RS.
eluted.
B. Gives the reactions of sulphates (2.3.1).
Inject the test solution and proceed with gradient elution,
Tests pumping the mobile phase mixture under the conditions
mentioned above for about 80 minutes or until the
pH (2.4.24). 4.5 to 6.0, determined in a solution containing 10 demethylbleomycin A2 is eluted. The usual order of elution is
Units per ml. bleomycinic acid, bleomycin A2 (first principal peak), bleomycin
A5, bleomycin B2 (second principal peak), bleomycin B4 and
Copper. Not more than 0.02 per cent, determined by Method A
demethylbleomycin A2 (retention time relative to bleomycin
or Method B
A2, between 1.5 and 2.5).
A. Weigh accurately a quantity containing about 50 mg of
Measure the peak responses of all the peaks. Calculate the
bleomycin, transfer to a 60-ml separator and dissolve in 10.0
percentage contents of each bleomycin component by
ml of 0.1 M hydrochloric acid. Add 10 ml of a 0.01 per cent
comparing the ratios of the individual areas of the peaks with
w/v solution of zinc bis (diphenyl dithiocarbamate) in carbon
that of the total area of all bleomycins.
tetrachloride and shake vigorously for 1 minute. Allow the
layers to separate, filter the lower layer through 1 g of Bacterial endotoxins (2.2.3). Not more than 10.0 Endotoxin
anhydrous sodium sulphate. Treat similarly 10.0 ml of copper Units per unit of bleomycin.
standard solution (10 ppm Cu) and measure the absorbances
(2.4.7) of the two solutions at the maximum at about 435 nm,
by drying the combined contents of two containers in an
using carbon tetrachloride as the blank.
oven at 60° at a pressure not exceeding 0.7 kPa for 3 hours.
B. Determine by atomic absorption spectrophotometry (2.4.2)
Assay. Determine the weight of the contents of 10 containers.
measuring at 324.7 nm using an air-acetylene flame and a
Mix the contents of the containers and determine by the
solution prepared in the following manner: Weigh accurately
microbiological assay of antibiotics, Method A or B (2.2.10)
a quantity containing about 75 mg of bleomycin, dissolve in
and express the results in Units per vial.
water and dilute to 10 .0 ml with the same solvent. Use copper
solution AAS suitably diluted with water, for preparing the Storage. The sealed container should be protected from
reference solutions. light.
193
BORIC ACID IP 2007
Labelling. The label states the total number of units contained Bromhexine Hydrochloride
in the sealed container.
Br
Boric Acid N
,HCl
CH3
H3BO3 Mol. Wt. 61.8 NH2
Boric Acid contains not less than 99.5 per cent and not more Br
than 100.5 per cent of H3BO3, calculated on the dried basis.
Description. A white, crystalline powder or colourless shiny C14H20Br2N2,HCl Mol. Wt. 412.6
plates unctuous to the touch or white crystals; odourless. Bromhexine Hydrochloride is 2-amino-3,5-
dibromobenzyl(cyclohexyl)methylamine hydrochloride.
Identification
Bromhexine Hydrochloride contains not less than 98.5 per
A. Dissolve 0.1 g by gently warming with 5 ml of methanol to cent and not more than 101.5 per cent of C14H20Br2N2,HCl,
which a few drops of sulphuric acid have been added. Ignite calculated on the dried basis.
the solution; the flame has a green border.
Description. A white or almost white, crystalline powder;
B. Dissolve 3.0 g in 90 ml of boiling distilled water, cool; the odourless or almost odourless.
solution is faintly acid.
Identification
Tests
Test A may be omitted if tests B, C, D and E are carried out.
Appearance of solution. A 3.5 per cent w/v solution in boiling Tests B, C and D may be omitted if tests A and E are carried
water is clear (2.4.1), and colourless (2.4.1). out.
pH (2.4.24). 3.8 to 4.8, determined in the solution obtained in A. Determine by infrared absorption spectrophotometry (2.4.6).
Identification test B. Compare the spectrum with that obtained with bromhexine
Solubility in ethanol. Dissolve 1.0 g in 10 ml of boiling ethanol hydrochloride RS or with the reference spectrum of
(95 per cent); the solution is not more opalescent than bromhexine hydrochloride.
opalescence standard OS2 (2.4.1), and colourless (2.4.1). B. In the test for Related Substances, the principal spot in the
Arsenic (2.3.10). Dissolve 1.0 g in 50 ml of water containing chromatogram obtained with test solution (b) corresponds to
2 g of citric acid and add 0.1 ml of stannous chloride solution that in the chromatogram obtained with reference solution (c).
AsT and 10 ml of hydrochloric acid. The resulting solution C. Dissolve about 25 mg in a mixture of 1 ml of 1 M sulphuric
complies with the limit test for arsenic (10 ppm). acid and 50 ml of water, add 2 ml of dichloromethane and 5 ml
Heavy metals (2.3.13). A solution produced by dissolving of a freshly prepared 2 per cent w/v solution of chloramine T
1.0 g in 2 ml of dilute acetic acid and diluting with sufficient and shake; a brownish yellow colour is produced in the lower
water to produce 25 ml complies with the limit test for heavy layer.
metals, Method A (20 ppm). D. A solution prepared by dissolving about 1 mg in 3 ml of 0.1
Sulphates (2.3.17). Dissolve 0.33 g in 10 ml of boiling water M hydrochloric acid gives the reaction for primary aromatic
and dilute to 15 ml with water. The solution complies with the amines (2.3.1).
limit test for sulphates (450 ppm). E. Dissolve about 20 mg in 1 ml of methanol and add 1 ml of
Loss on drying (2.4.19). Not more than 0.5 per cent, determined water. The solution gives reaction A of chlorides (2.3.1).
on 1.0 g by drying over silica gel for 5 hours.
Tests
Assay. Weigh accurately about 2.0 g, dissolve in a mixture of
50 ml of water and 100 ml of glycerin, previously neutralised Related substances. Determine by thin-layer chromatography
to phenolphthalein solution. Titrate with 1 M sodium (2.4.17), coating the plate with silica gel GF254.
hydroxide using phenolphthalein solution as indicator. Mobile phase. A mixture of 66 volumes of 1-butanol, 17
1 ml of 1 M sodium hydroxide is equivalent to 0.06183 g of volumes of glacial acetic acid and 17 volumes of water.
H3BO3. Test solution (a). Dissolve 2.0 g of the substance under
Labelling. The label states that it is not meant for internal use. examination in 100 ml of methanol.
194
IP 2007 BROMHEXINE TABLETS
Test solution (b). Dilute 10 ml of test solution (a) to 100 ml crystallisation. Mix the residue with 1 g of sodium carbonate,
with methanol. heat at a dull red heat for 10 minutes, allow to cool, extract with
Reference solution (a). Dilute 2.5 ml of test solution (b) to 100 water and filter. The filtrate, after acidification with 2 M nitric
ml with methanol. acid, yields reaction A of bromides (2.3.1).
C. Shake a quantity of the powdered tablets containing 20 mg
Reference solution (b). Dilute 15 ml of reference solution (a)
of Bromhexine Hydrochloride with 10 ml methanol and filter.
to 20 ml with methanol and mix.
The filtrate gives reaction A of chlorides (2.3.1).
bromhexine hydrochloride RS in methanol. Tests
Apply to the plate in small quantities a total of 20 µl of each Related substances. Determine by thin-layer chromatography
solution After development, dry the plate in air and examine in (2.4.17), coating the plate with silica gel G.
ultraviolet light at 254 nm. Any secondary spot in the
chromatogram obtained with test solution (a) is not more Mobile phase. A mixture of 90 volumes of heptane and 10
intense than the spot in the chromatogram obtained with volumes of ethanol.
reference solution (a). The test is not valid unless there is a
clearly visible spot in the chromatogram obtained with
containing 0.1 g of Bromhexine Hydrochloride with 10 ml of
reference solution (b).
methanol for 5 minutes, centrifuge and mix 9 volumes of the
Sulphated ash (2.3.18). Not more than 0.1 per cent. supernatant liquid with 1 volume of strong ammonia solution.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Reference solution. Dilute 1 volume of the test solution to
on 1.0 g by drying in an oven at 105°. 40 volumes with methanol.
Assay. Weigh accurately about 0.3 g, dissolve in 70 ml of Apply to the plate 20µI of each solution. After development,
ethanol (95 per cent), add 1 ml of 0.1 M hydrochloric acid dry the plate in a current of warm air, place in a tank containing
and titrate with 0.1 M sodium hydroxide, determining the end- a freshly prepared 10 per cent w/v solution of sodium nitrite
point potentiometrically (2.4.25). Record the volume added in 5 M hydrochloric acid and allow to stand for 1 minute.
between the two inflections. Remove the plate and immediately spray with a 0.5 per cent
1ml of 0.1 M sodium hydroxide is equivalent to 0.04126 g of w/v solution of N-(1-naphthyl)ethylenediamine
C14H20Br2N2,HCl. dihydrochloride in methanol. Any secondary spot in the
Storage. Store protected from light. intense than the spot in the chromatogram obtained with the
reference solution.
Bromhexine Tablets Assay. Weigh and powder 20 tablets. Weigh accurately a
quantity of the powder containing about 8 mg of Bromhexine
Bromhexine Hydrochloride Tablets Hydrochloride, shake with 50 ml of 0.1 M methanolic
Bromhexine Tablets contain not less than 92.5 per cent and hydrochloric acid for 30 minutes, add sufficient 0.1 M
not more than 107.5 per cent of the stated amount of methanolic hydrochloric acid to produce 100.0 ml and filter.
bromhexine hydrochloride, C14H20Br2N2,HCl. Measure the absorbance of the filtrate at the maximum at about
317 nm (2.4.7). Calculate the content of C14H20Br2N2,HCl taking
Identification 87 as the specific absorbance at 317 nm.
solution obtained in the Assay shows an absorption maximum
only at about 317 nm.
B. Suspend a quantity of the powdered tablets containing 0.1
g of Bromhexine Hydrochloride in 5 ml of dilute ammonia
solution and extract with two quantities, each of 20 ml, of
chloroform. Wash the combined extracts with 5 ml of water,
filter through anhydrous sodium sulphate and evaporate the
filtrate to dryness using a rotary evaporator. If necessary,
scratch the inside of the flask with a glass rod to induce
195
BROMOCRIPTINE MESYLATE IP 2007
Bromocriptine Mesylate barium chloride to the filtrate; it remains clear. Mix another
0.1 g with 0.5 g of anhydrous sodium carbonate and ignite
until a white residue is obtained. After cooling, dissolve the
residue in 5 ml of water (solution A); solution A gives the
CH3 OH reactions of sulphates (2.3.1).
H3 C O
H E. Solution A gives reaction A of bromides (2.3.1).
HN
O O ,CH3SO3H Tests
O CH3
Appearance of solution. A 1.0 per cent w/v solution in
N CH CH3 methanol is clear (2.4.1), and not more intensely coloured
3
H than reference solution BS5, YS5 or BYS5 (2.4.1).
HN in a mixture of 2 volumes of methanol and 8 volumes of water.
Br Specific optical rotation (2.4.22). + 95° to + 105°, determined
in a 1.0 per cent w/v solution in a mixture of equal volumes of
methanol and dichloromethane.
C32H40BrN5O5,CH4O3S Mol. Wt. 750.7
Bromocriptine Mesylate is (5'S)-2-bromo-12'-hydroxy-2'-(1- (2.4.17), coating the plate with silica gel G.
methylethyl)-5'-(2-methylpropyl)ergotaman-3',6',18-trione
methanesulphonate Mobile phase. A mixture of 100 volumes of ether, 88 volumes
of dichloromethane, 3 volumes of 2-propanol, 1.5 volumes
Bromocriptine Mesylate contains not less than 98.0 per cent of water and 0.1 volume of strong ammonia solution.
and not more than 101.0 per cent of C32H40BrN5O5,CH4O3S,
calculated on the dried basis. Test solution (a). Prepare freshly a solution containing 1 g of
the substance under examination in 100 ml of a mixture of
Description. A white or slightly coloured, fine crystalline 4 volumes of dichloromethane, 3 volumes of ethanol (95 per
powder; very sensitive to light. cent) and 3 volumes of methanol.
NOTE — Carry out the tests as rapidly as possible without Test solution (b). Freshly dilute 10 ml of test solution (a) to
exposure to daylight and with minimum exposure to artificial 100 ml with the same solvent mixture.
light.
Reference solution (a). Dilute 2 ml of test solution (b) to 50 ml
Identification with the same solvent mixture.
Reference solution (b). Dilute 10 ml of reference solution (a)
to 20 ml with the same solvent mixture.
Tests B, C, D and E may be omitted if test A is carried out.
Reference solution (c). Dilute 10 ml of reference solution (b)
A. Determine by infrared absorption spectrophotometry in a
to 20 ml with the same solvent mixture.
mineral oil dispersion (2.4.6). Compare the spectrum with that
obtained with bromocriptine mesylate RS or with the reference Reference solution (d). A 0.1 per cent w/v solution of
spectrum of bromocriptine mesylate. bromocriptine mesylate RS in the same solvent mixture.
B. Dissolve 5 mg in 5 ml of methanol and dilute to 100 ml with Apply to the plate, as 1-cm bands, 10 µl of each solution.
0.01 M hydrochloric acid. The resulting solution, when Apply test solution (a) to the plate as the last solution and
examined in the range 230 nm to 360 nm (2.4.7) shows an develop the chromatogram immediately in an unsaturated tank.
absorption maximum at about 305 nm and a minimum at about After development, allow the plate to dry in a current of cold
270 nm; absorbance at about 305 nm, 0.60 to 0.68. air, spray with ethanolic ammonium molybdate solution and
heat at 100° until bands are visible (about 10 minutes). Any
C. In the test for Related substances, the principal band in the
secondary band in the chromatogram obtained with test
chromatogram obtained with test solution (b) corresponds to
solution (a) is not more intense than the principal band in the
chromatogram obtained with reference solution (a) and not
(d).
more than one such band is more intense than the principal
D. To about 0.1 g add 5 ml of 2 M hydrochloric acid, shake for band in the chromatogram obtained with reference solution
5 minutes, filter and add 1 ml of a 6 per cent w/v solution of (b) and not more than one other such band is more intense
196
IP 2007 BROMOCRIPTINE CAPSULES
than the principal band in the chromatogram obtained with Test solution (a). Shake a quantity of the contents of the
solution (c). capsules containing 20 mg of bromocriptine with 10 ml of
methanol for 20 minutes and centrifuge.
methanol.
80° at a pressure of 1.5 to 2.5 kPa for 5 hours. Reference solution (a). Dilute 3 ml of test solution (a) to
Assay. Weigh accurately about 0.5 g, dissolve in 80 ml of a
mixture of 10 volumes of anhydrous glacial acetic acid and Reference solution (b). Dilute 1ml of test solution (a) to 100 ml
70 volumes of acetic anhydride. Titrate with 0.1 M perchloric with methanol.
acid, determining the end-point potentiometrically (2.4.25) Reference solution (c). Dilute 1 ml of test solution (a) to
Carry out a blank titration. 200 ml with methanol.
1 ml of 0.1 M perchloric acid is equivalent to 0.07507 g of Reference solution (d). A 0.023 per cent w/v solution of
C32H40BrN5O5,CH4O3S. bromocriptine mesylate RS in methanol.
Storage. Store protected from light in a deep freezer Apply to the plate, as 1-cm bands, 50 µl of each solution.
(temperature not exceeding –15°). Apply test solution (a) to the plate as the last solution and
develop the chromatogram immediately in an unsaturated tank.
After development, allow the plate to dry in a current of cold
air, spray with ethanolic ammonium molybdate solution and
Bromocriptine Capsules heat at 100° until bands are visible (about 10 minutes). In the
chromatogram obtained with test solution (a) any secondary
Bromocriptine Mesylate Capsules band is not more intense than the band in the chromatogram
Bromocriptine Capsules contain not less than 90.0 per cent obtained with reference solution (a). Not more than one such
and not more than 110.0 per cent of the stated amount of band is more intense than the band in the chromatogram
bromocriptine, C32H40BrN5O5. obtained with reference solution (b) and not more than two
further such bands are more intense than the band in the
NOTE — Carry out the tests as rapidly as possible without chromatogram obtained with reference solution (c). Disregard
exposure to daylight and with minimum exposure to artificial any band within 20 mm of the line of application.
light.
Identification Capsules.
Empty the contents of one capsule, crush, if necessary, add
A. Shake a quantity of the contents of the capsules containing 10.0 ml of methanol, shake vigorously and centrifuge. If
10 mg of bromocriptine with 50 ml of methanol for 30 minutes, necessary, dilute the solution appropriately and carry out the
centrifuge and dilute 5 ml of the supernatant liquid to 20 ml procedure described under the Assay beginning at the words
with methanol. When examined in the range 230 nm to 360 nm “measure the absorbance.....”.
(2.4.7), the resulting solution shows an absorption maximum
at about 305 nm and a minimum at about 270 nm. Other tests. Comply with the tests stated under Capsules.
B. In the test for Related substances, the principal band in the Assay. Weigh accurately a quantity of the mixed contents of
chromatogram obtained with test solution (b) corresponds to 20 capsules containing about 25 mg of bromocriptine and
that in the chromatogram obtained with reference solution shake vigorously with 30 ml of methanol. Dilute to 100.0 ml
(d). with methanol and filter. Dilute further with methanol to yield
a final concentration of about 50 mcg per ml and measure the
Tests absorbance of the resulting solution at the maximum at about
305 nm (2.4.7). Calculate the content of C32H40BrN5O5 from the
Related substances. Determine by thin-layer chromatography absorbance obtained by repeating the operation using
(2.4.17), coating the plate with silica gel G. bromocriptine mesylate RS equivalent to 25 mg of
Mobile phase. A mixture of 100 volumes of ether, 88 volumes bromocriptine instead of the substance under examination.
of dichloromethane, 3 volumes of 2-propanol, 1.5 volumes Storage. Store protected from light.
of water and 0.1 volume of strong ammonia solution.
Prepare the following solutions freshly. equivalent amount of bromocriptine.
197
BROMOCRIPTINE TABLETS IP 2007
Bromocriptine Tablets Reference solution (b). Dilute 1ml of test solution (b) to 10 ml
Bromocriptine Mesylate Tablets Reference solution (c). Dilute 1 ml of test solution (b) to
Bromocriptine Tablets contain not less than 90.0 per cent and 20 ml with the same solvent mixture.
not more than 110.0 per cent of the stated amount of Reference solution (d). A 0.055 per cent w/v solution of
bromocriptine, C32H40BrN5O5. bromocriptine mesylate RS in the same solvent mixture.
NOTE — Carry out the tests as rapidly as possible without Apply to the plate, as 1-cm bands, 20 µl of each solution.
exposure to daylight and with minimum exposure to artificial Apply test solution (a) to the plate as the last solution and
light. develop the chromatogram immediately in an unsaturated tank.
After development, allow the plate to dry in a current of cold
Identification
air, spray with ethanolic ammonium molybdate solution and
A. Shake a quantity of powdered tablets containing about heat at 100° until bands are visible (about 10 minutes). Any
20 mg of bromocriptine with 20 ml of methanol, filter, evaporate secondary band in the chromatogram obtained with test
the filtrate to dryness on a water-bath and dry at 105° for solution (a) is not more intense than the band in the
1 hour.The residue complies with the following test. chromatogram obtained with reference solution (a). Not more
Determine by infrared absorption spectrophotometry (2.4.6). than one such band is more intense than the band in the
Compare the spectrum with that obtained with bromocriptine chromatogram obtained with reference solution (b) and not
mesylate RS or with the reference spectrum of bromocriptine more than a further two such bands are more intense than the
mesylate. band in the chromatogram obtained with solution (c). Disregard
any band within 20 mm of the line of application.
B. Shake a quantity of the powdered tablets containing about
10 mg of bromocriptine with 50 ml of methanol for 30 minutes, Uniformity of content. Comply with the test stated under
centrifuge and dilute 5 ml of the supernatant liquid to 20 ml Tablets.
with methanol. When examined in the range 230 nm to 360 nm Finely crush one tablet, add 10.0 ml of methanol, shake
(2.4.7), the resulting solution shows an absorption maximum vigorously and centrifuge. If necessary, dilute the solution
at about 305 nm and a minimum at about 270 nm. appropriately and carry out the procedure described under
C. In the test for Related substances, the principal band in the the Assay beginning at the words “Measure the
chromatogram obtained with test solution (b) corresponds to absorbance.....”.
that in the chromatogram obtained with reference solution Other tests. Comply with the tests stated under Tablets.
(d). Assay. Weigh and powder 20 tablets. Weigh accurately a
Tests quantity of the powder containing about 2.5 mg of
bromocriptine and shake vigorously with 30 ml of methanol.
Related substances. Determine by thin-layer chromatography Dilute to 50.0 ml with methanol and filter. Measure the
(2.4.17), coating the plate with silica gel G. absorbance of the resulting solution at the maximum at about
Mobile phase. A mixture of 100 volumes of ether, 88 volumes 305 nm (2.4.7). Calculate the content of C32H40BrN5O5 from the
of dichloromethane, 3 volumes of 2-propanol, 1.5 volumes absorbance obtained by repeating the operation with
of water and 0.1 volume of strong ammonia solution. bromocriptine mesylate RS equivalent to 25 mg of
bromocriptine in 50 ml methanol and diluting 5.0 ml to 50.0 ml
Prepare the following solutions freshly. with methanol.
Test solution (a). Shake a quantity of the powdered tablets Storage. Store protected from light.
containing 10 mg of bromocriptine with 25 ml of a mixture of
equal volumes of chloroform and methanol for 30 minutes, Labelling. The label states the strength in terms of the
filter through a sintered glass filter (porosity No. 4) and wash equivalent amount of bromocriptine.
the residue with two quantities, each of 5 ml, of the same
solvent mixture. Evaporate the filtrate and washings to dryness
at 25° at a pressure of 2 kPa, dissolve the residue in 2 ml of the Bronopol
same solvent mixture and centrifuge.
B r NO 2
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with HO OH
the same solvent mixture.
C3H6BrNO4 Mol. Wt. 200.0
Reference solution (a). Dilute 3 ml of test solution (a) to
100 ml with the same solvent mixture. Bronopol is 2-bromo-2-nitropropane-1,3-diol.
198
IP 2007 BUDESONIDE
Bronopol contains not less than 99.0 per cent and not more Inject the test solution, reference solution (a) and reference
than 101.0 per cent of C3H6BrNO4, calculated on the anhydrous solution (b). Continue the chromatography for 3 times the
basis. retention time of the principal peak. In the chromatogram
Description. White or almost white crystals or crystalline obtained with the test solution the area of any peaks
powder. corresponding to 2-methyl-2-nitropropane-1,3-diol and tris
(hydroxymethyl) nitromethane are not more than the area of
Identification the corresponding peaks in the chromatogram obtained with
reference solution (b) (0.5 per cent each) and the area of any
A. Determine by infrared absorption spectrophotometry (2.4.6). other secondary peak is not more than the area of the principal
Compare the spectrum with that obtained with bronopol RS. peak in the chromatogram obtained with reference solution
(b).
B. Dissolve 0.1 g in 10 ml of water, add 10 ml of 7.5 M sodium
hydroxide and carefully with constant stirring and cooling, Sulphated ash (2.3.18). Not more than 0.1 per cent.
0.5 g of nickel-aluminium alloy. Allow the reaction to subside, Water (2.3.43). Not more than 0.5 per cent, determined on
filter and carefully neutralise with nitric acid. The resulting 5.0 g.
solution gives reaction A of bromides (2.3.1).
Assay. In a flask fitted with a reflux condenser dissolve 0.4 g
Tests in 15 ml of water and add 15 ml of 7.5 M sodium hydroxide.
Slowly, with caution, add 2 g of nickel-aluminium alloy
pH (2.4.24). 5.0 to 7.0, determined on 1.0 per cent w/v solution. through the reflux condenser, agitating the flask whilst cooling
Related substances. Determine by liquid chromatography under running water. Allow the mixture to stand for 10 minutes
(2.4.14) and boil for 1 hour. Cool and filter under reduced pressure,
washing the condenser, flask and residue with 150 ml of water.
Test solution. Dissolve 0.2 g of the substance under Combine the filtrate and washings, add 25 ml of nitric acid
examination in 100 ml of mobile phase. and 40 ml of 0.1 M silver nitrate, shake vigorously and titrate
Reference solution (a). Dilute 5 ml of the test solution to 50 ml with 0.1 M ammonium thiocyanate using ammonium iron(III)
with the mobile phase. Further, dilute 1 ml of the solution to sulphate solution as indicator. Carry out a blank titration.
100 ml with the mobile phase. 1 ml of 0.1 M silver nitrate is equivalent to 0.020 g of
Reference solution (b). A solution containing 0.001 per cent C3H6BrNO4.
w/v each of 2-methyl-2-nitropropan-1,3-diol and Storage. Store protected from light.
tris(hydroxymethyl)nitromethane in the mobile phase.
Reference solution (c). A solution containing 0.0002 per cent
w/v each of 2-methyl-2-nitropropane-1,3-diol, 2-
nitroethanol, sodium bromide and tris(hydroxymethyl)- Budesonide
nitromethan in the mobile phase.
Chromatographic system OH
– a stainless steel column 15 cm x 4.6 mm, packed with O
octadecylsilane bonded to porous silica (5 µm), H3C
HO O CH3
– temperature 35°,
– mobile phase: a mixture of 189 volumes of water , H3C H O
H
10 volumes of acetonitrile and 1 volume of a 10 per
H H
cent v/v solution of orthophosphoric acid, adjusting
the pH to 3.0 using 2 M sodium hydroxide, O
– spectrophotometer set at 214 nm, C25H34O6 Mol. Wt. 430.5
– a 20 µl loop injector. Budesonide is a mixture of the C-22S (epimer A) and the C-
Inject reference solution (c).The test is not valid unless the 22R (epimer B) epimers of 16α,17-[(1RS)-
resolution between the peaks corresponding to sodium butylidenebis(oxy)]-11β,21-dihydroxypregna-1,4-diene-3,20-
bromide and tris (hydroxymethyl)nitromethane is at least dione.
1.0 and the resolution between the peaks corresponding to Budesonide contains not less than 98.0 per cent and not more
tris(hydroxymethyl)nitromethane and 2-nitroethanol is at than 102.0 per cent of a mixture of epimers A and B, C25H34O6,
least 1.5. calculated on the dried basis.
199
BUDESONIDE IP 2007
Description. A white or almost white, crystalline powder. pH 3.2 to 100 ml and allow to stand for at least 15 minutes
before use and filter.
Identification
Reference solution (a). Dissolve 50 mg of budesonide RS in
Test A may be omitted if tests B, C and D are carried out. Tests 30 ml of acetonitrile. Add about 60 ml of phosphate buffer pH
B, C and D may be omitted if test A is carried out. 3.2 and disperse, if necessary, with the aid of ultrasound to
A. Determine by infrared absorption spectrophotometry (2.4.6). dissolve. Dilute to 100 ml with phosphate buffer pH 3.2 and
Compare the spectrum with that obtained with budesonide allow to stand for at least 15 minutes before use and filter.
RS or with the reference spectrum of budesonide. Reference solution (b). Dilute reference solution (a) with the
B. Determine by thin-layer chromatography (2.4.17), coating mobile phase to get a 0.00025 per cent w/v solution of
the plate with silica gel GF254. budesonide.
Mobile phase. Add a mixture of 1.2 volumes of water and 8 Use the chromatographic system described in the Assay.
volumes of methanol to a mixture of 15 volumes of ether and Inject reference solution (a). The test is not valid unless the
77 volumes of dichloromethane. resolution between epimer B peak and epimer A peak is not
Solvent mixture. 1 volume of methanol and 9 volumes of less than 1.5, the tailing factor for epimer B peak is not more
methylene chloride. than 1.5 and the relative standard deviation of sum of epimer
A and epimer B peaks for replicate injections is not more than
2.0 per cent.
chromatogram obtained with the test solution: the area of any
budesonide RS in the solvent mixture.
peak, other than the principal peak, is not greater than the area
Reference solution (b). A solution containing 0.25 per cent w/ of the principal peak in the chromatogram obtained with
v of triamcinolone acetonide RS and 0.25 per cent w/v of reference solution (b) (0.5 per cent) and the sum of the areas
budesonide RS in the solvent mixture. of all the peaks, other than the principal peak, is not greater
Apply to the plate 5 µl of each solution. After development, than thrice the area of the principal peak in the chromatogram
dry the plate in air and examine in ultraviolet light at 254 nm. obtained with reference solution (b) (1.5 per cent).
The principal spot in the chromatogram obtained with the test Loss on drying (2.4.19). Not more than 0.5 per cent, determined
solution corresponds to the principal spot in the chromatogram on 1 g by drying in an oven at 105º.
obtained with reference solution (a). The test is not valid unless
the chromatogram obtained with reference solution (b) shows Assay. Determine by liquid chromatography (2.4.14).
two clearly separated spots. Test solution. Dissolve 50 mg of the substance under
C. Dissolve about 2 mg in 2 ml of sulphuric acid. A yellow examination in 30 ml of acetonitrile and dilute to 100.0 ml with
colour appears in 5 minutes and the colour changes to brown phosphate buffer solution pH 3.2 and filter.
or reddish-brown in 30 minutes. Add cautiously the solution Reference solution. Dissolve 50 mg of budesonide RS in 30 ml
to 10 ml of water and mix. The colour fades and a clear solution of acetonitrile and dilute to 100.0 ml with phosphate buffer
remains. solution pH 3.2.
D. Dissolve about 1 mg in 2 ml of a solution containing 2 g of Chromatographic system
phosphomolybdic acid in a mixture of 10 ml of dilute sodium – a stainless steel column 15 cm x 4.6 mm, packed with
hydroxide solution, 15 ml of water and 25 ml of glacial acetic octadecylsilyl silica (5 µm),
acid. Heat for 5 minutes on a water-bath. Cool in iced water for – mobile phase: a mixture of 34 volumes of acetonitrile
10 minutes and add 3 ml of dilute sodium hydroxide solution. and 66 volumes of a buffer solution prepared by adding
The solution turns blue. 100 ml of 0.25 per cent w/v solution of orthophosphoric
acid to 900 ml of 0.4 per cent w/v solution of sodium
Tests dihydrogen phosphate and adjusting the pH to 3.2, if
Related substances. Determine by liquid chromatography necessary.
(2.4.14). – flow rate. 1.5 ml per minute,
examination in 30 ml of acetonitrile. Add about 60 ml of
phosphate buffer pH 3.2 and, if necessary, disperse with the Inject the reference solution. The test is not valid unless the
aid of ultrasound to dissolve. Dilute with phosphate buffer resolution between epimer B peak and epimer A peak is not
200
IP 2007 BUPIVACAINE HYDROCHLORIDE
less than 1.5, the tailing factor for epimer B peak is not more D. In the test for Related substances, the principal spot in the
than 1.5, the column efficiency determined for epimer B peak chromatogram obtained with test solution (b) corresponds to
is not less than 4000 theoretical plates and the relative standard that in the chromatogram obtained with reference solution
deviation for the sum of epimer A and B peaks for replicate (b).
injections is not more than 2.0 per cent. E. A 10 per cent w/v solution gives reaction A of chlorides
Inject the test solution and the reference solution. (2.3.1).
Calculate the content of C25H34O6. Tests
Acidity or alkalinity. To 10 ml of a 2.0 per cent w/v solution in
carbon dioxide-free water add 0.2 ml of 0.01 M sodium
hydroxide; the pH is not less than 4.7. Add 0.4 ml of 0.01 M
hydrochloric acid; the pH is not more than 4.7 (2.4.24).
Bupivacaine Hydrochloride
dioxide-free water is clear (2.4.1), and colourless (2.4.1).
H3 C H3C
O
,HCl,H2O (2.4.17), coating the plate with silica gel G.
N
N Mobile phase. A mixture of 100 volumes of methanol and
H CH3 0.1 volume of strong ammonia solution.
C18H28N2O,HCl,H2O Mol. Wt. 342.9 examination in 100 ml of methanol.
Bupivacaine Hydrochloride is 1-butyl-N-(2,6-dimethylphenyl)- Test solution (b). Dilute 10 ml of test solution (a) to 100 ml
2-piperidinecarboxamide hydrochloride monohydrate. with methanol.
Bupivacaine Hydrochloride contains not less than 98.5 per Reference solution (a). Dilute 5 ml of test solution (b) to
cent and not more than 101.0 per cent of C18H28N2O,HCl, 100 ml with methanol.
calculated on the dried basis.
Description. A white, crystalline powder or colourless crystals; bupivacaine hydrochloride RS in methanol.
almost odourless.
Identification dry the plate in air and spray with dilute potassium
Test A may be omitted if tests B, C, D and E are carried out. chromatogram obtained with test solution (a) is not more
Tests B, C and D may be omitted if tests A and E are carried intense than the spot in the chromatogram obtained with
out. reference solution (a).
A. Determine by infrared absorption spectrphotometry (2.4.6). 2,6-Dimethylaniline. To 2.0 ml of a 5.0 per cent w/v solution
Compare the spectrum with that obtained with bupivacaine in methanol (solution A) add 1 ml of a freshly prepared 1 per
hydrochloride RS or with the reference spectrum of cent w/v solution of 4-dimethylaminobenzaldehyde in
bupivacaine hydrochloride. methanol and 2 ml of glacial acetic acid and allow to stand
for 10 minutes. Any yellow colour produced is not more intense
than that obtained with a solution prepared at the same time
and in the same manner using 2 ml of a 0.0005 per cent w/v
two absorption maxima at about 263 nm and 271 nm;
solution of 2,6-dimethylaniline in methanol in place of
absorbance at about 263 nm, about 0.70 and at about 271 nm,
solution A (100 ppm).
about 0.57.
Heavy metals (2.3.13). A 10.0 per cent w/v solution in a mixture
C. Dissolve 0.1 g in 10 ml of water, add 2 ml of 2 M sodium
of 85 volumes of methanol and 15 volumes of water complies
hydroxide and shake with two quantities, each of 15 ml, of
with the limit test for heavy metals Method D (10 ppm).Prepare
ether. Dry the combined ether extracts over anhydrous sodium
the standard using lead standard solution (1 ppm Pb) obtained
sulphate, filter, evaporate the ether, recrystallise the residue
by diluting lead standard solution (20 ppm Pb) with a mixture
from ethanol (90 per cent) and dry the residue at a pressure
of 85 volumes of methanol and 15 volumes of water.
of 1.5 to 2.5 kPa. The melting range (2.4.21) of the residue is
between 105° and 108° (2.4.21). Sulphated ash (2.3.18). Not more than 0.1 per cent.
201
BUPIVACAINE INJECTION IP 2007
Loss on drying (2.4.19). 4.5 to 6.0 per cent, determined on Mobile phase. A mixture of 100 volumes of methanol and
1.0 g by drying in an oven at 105°. 0.1 volume of strong ammonia solution.
Assay. Weigh accurately about 0.25 g, dissolve in a mixture of Test solution. Evaporate almost to dryness a volume
5.0 ml of 0.01 M hydrochloric acid and 50 ml of ethanol containing 0.1 g of anhydrous bupivacaine hydrochloride
(95 per cent) and titrate with 0.01 M ethanolic sodium using a rotary evaporator, add sufficient methanol to the
hydroxide, determining the end-point potentiometrically residue to produce 2 ml, mix well, centrifuge and use the
(2.4.25). Note the volume added between the inflections. supernatant liquid.
1 ml of 0.01 M ethanolic sodium hydroxide is equivalent to Reference solution. Dilute 1 volume of the test solution to
0.03249 g of C18H28N2O,HCl. 100 volumes with methanol.
Storage. Store protected from light. Apply to the plate 10 µl of each solution. After development,
dry the plate in air and spray with dilute potassium
Bupivacaine Injection intense than the spot in the chromatogram obtained with the
reference solution.
Bupivacaine Hydrochloride Injection
2,6-Dimethylaniline. To a volume containing 25 mg of
Bupivacaine Injection is a sterile solution of Bupivacaine anhydrous bupivacaine hydrochloride add water, if necessary,
Hydrochloride in Water for Injection. to produce 10 ml and sufficient 2 M sodium hydroxide to
Bupivacaine Injection contains not less than 92.5 per cent make the solution just alkaline. Extract with three quantities,
and not more than 107.5 per cent of the stated amount of each of 5 ml, of chloroform. Dry the combined extracts over
anhydrous bupivacaine hydrochloride, C18H28N2O,HCl. anhydrous sodium sulphate, filter, wash the filter with 5 ml of
chloroform and evaporate the filtrate to dryness using a rotary
Identification evaporator. Dissolve the residue in 2 ml of methanol. Add 1 ml
of a freshly prepared 1 per cent w/v solution of
A. To a volume containing 25 mg of anhydrous bupivacaine 4-dimethylaminobenzaldehyde in methanol and 2 ml of
hydrochloride add 2 ml of strong ammonia solution, shake glacial acetic acid and allow to stand for 10 minutes. Any
and filter. Wash the precipitate with water and dry at 60° at a yellow colour produced is not more intense than that obtained
pressure of 2 kPa for 16 hours. The residue complies with the with a solution prepared at the same time and in the same
following test. manner using 2 ml of a 0.0005 per cent w/v solution of 2,6-
Determine by infrared absorption spectrophotometry (2.4.6). dimethylaniline in methanol in place of the injection under
Compare the spectrum with that obtained with bupivacaine examination.
hydrochloride RS treated in the same manner or with the Other tests. Complies with the tests stated under Parenteral
reference spectrum of bupivacaine. Preparations (Injections).
B. To a volume containing 50 mg of anhydrous bupivacaine Assay. To an accurately measured volume containing about
hydrochloride add 15 ml of picric acid solution; the precipitate, 0.5 g of anhydrous bupivacaine hydrochloride add 5 ml of
after rapid washing with a small quantity of water followed by water and 2 ml of 1 M sodium hydroxide and extract with
successive quantities, each of 2 ml, of methanol and ether three quantities, each of 15 ml, of chloroform. Combine the
melts at about 194° (2.4.21). chloroform extracts, wash with two quantities, each of 5 ml, of
C. To a volume containing 50 mg of anhydrous bupivacaine water, extract the aqueous solutions with 5 ml of chloroform
hydrochloride add 2 ml of a 10 per cent w/v solution of and evaporate the combined chloroform extracts to dryness
disodium hydrogen phosphate and sufficient iodine solution on a water-bath. Add two successive quantities, each of 5 ml,
to produce a distinct brown colour. Remove the excess iodine of acetone and evaporate. Dissolve the residue in 50 ml of
by adding 0.1 M sodium thiosulphate; no pink colour is anhydrous glacial acetic acid. Add 15 ml of mercuric acetate
produced. solution. Titrate with 0.1 M perchloric acid, using crystal
violet solution as indicator. Carry out a blank titration.
Tests
pH (2.4.24). 4.0 to 6.5. C18H28N2O,HCl.
Related substances. Determine by thin-layer chromatography Storage. Store in single dose or multiple dose containers,
(2.4.17), coating the plate with silica gel G. preferably of Type 1 glass.
202
IP 2007 BUPRENORPHINE INJECTION
Labelling. The label states the strength in terms of the Mobile phase. A mixture of 85 volumes of toluene, 15 volumes
equivalent amount of anhydrous bupivacaine hydrochloride of methanol and 0.5 volume of strong ammonia solution.
in a suitable dose-volume. Prepare the following solutions freshly.
examination in 50 ml of methanol.
Buprenorphine Hydrochloride Reference solution. Dilute 1 ml of the test solution to 100 ml
with methanol. Mix well and dilute 10 ml of this solution to
HO
dry the plate in air and examine in ultraviolet light at 254 nm or
O expose to iodine vapours. Any secondary spot in the
, HCl chromatogram obtained with the test solution is not more
N
H3CO reference solution.
H
Heavy metals (2.3.13). Moisten the residue obtained in the
HO CH3 test for Sulphated ash with a few drops of hydrochloric acid
C(CH3)3
and evaporate almost to dryness on a water-bath. Dissolve
the residue in 10 ml of water by warming, cool, transfer to a
C29H41NO4,HCl Mol. Wt. 504.1 test-tube with the aid of 10 ml of water and add 2 ml of dilute
acetic acid. The solution complies with the limit test for heavy
Buprenorphine Hydrochloride is (6R,7R,14S)-17- metals, Method A (20 ppm). Prepare the standard using 5 ml of
cyclopropylmethyl-7,8-dihydro-7-[(1S)-1-hydroxy-1,2,2- lead standard solution (2 ppm Pb).
trimethylpropyl]-6-O-methyl-6,14-ethano-17-normorphine
hydrochloride. Sulphated ash (2.3.18). Not more than 0.2 per cent.
Buprenorphine Hydrochloride contains not less than 97.0 per Loss on drying (2.4.19). Not more than 1.0 per cent, determined
cent and not more than 102.0 per cent of C29H41NO4,HCl, on 0.5 g by drying in an oven at 105° for 4 hours.
calculated on the dried basis. Assay. Weigh accurately about 0.5 g, dissolve in 60 ml of
Description. A white to off-white, crystalline powder. anhydrous glacial acetic acid, add 10 ml of mercuric acetate
solution. Titrate with 0.1 M perchloric acid, using 0.1 ml of
Identification crystal violet solution as indicator to a green end-point. Carry
Compare the spectrum with that obtained with buprenorphine
C29H41NO4,HCl.
hydrochloride RS.
0.01 per cent w/v solution in 0.01 M hydrochloric acid shows Buprenorphine Injection
an absorption maximum at about 286 nm; absorbance at about
286 nm, about 0.33. Buprenorphine Hydrochloride Injection
C. Dissolve about 5 mg in 5 ml of hot water, add 2 ml of dilute Buprenorphine Injection is a sterile solution of Buprenorphine
hydrochloric acid and 2 ml of a 2 per cent w/v solution of Hydrochloride in Water for Injection.
sodium nitrite and allow to stand for 10 minutes; a yellow Buprenorphine Injection contains not less than 90.0 per cent
colour is produced. and not more than 110.0 per cent of the stated amount of
D. Dissolve 10 mg in 10 ml of hot water; add 2 ml of dilute buprenorphine, C29H41NO4.
nitric acid, shake and add 1 ml of silver nitrate solution; a
white precipitate is produced. Identification
Tests the plate with silica gel HF254.
Related substances. Determine by thin-layer chromatography Mobile phase. A mixture of 85 volumes of toluene, 15 volumes
(2.4.17), coating the plate with silica gel GF254. of methanol and 0.5 volume of strong ammonia solution.
203
BUPRENORPHINE TABLETS IP 2007
Test solution. Transfer a volume of the injection containing Buprenorphine Tablets

1.5 mg of Buprenorphine Hydrochloride to a 125-ml separator,
add 0.5 ml of dilute ammonia solution, shake and extract with Buprenorphine Hydrochloride Tablets
three quantities, each of 10 ml, of chloroform, washing each Buprenorphine Tablets contain not less than 90.0 per cent
chloroform extract with the same 10 ml of water and discard and not more than 110.0 per cent of the stated amount of
the water. Evaporate the combined chloroform extracts to buprenorphine, C29H41NO4.
dryness on a water-bath and dissolve the residue in 1.5 ml of
chloroform. Identification
Reference solution. Dissolve 1.5 mg of buprenorphine A. Determine by thin-layer chromatography (2.4.17), coating
hydrochloride RS in 5 ml of 0.01 M hydrochloric acid, transfer the plate with silica gel HF254.
the solution to a 125-ml separator and repeat the above
procedure beginning at the words “add 0.5 ml of dilute Mobile phase. A mixture of 85 volumes of toluene, 15 volumes
ammonia solution.....”. of methanol and 0.5 volume of strong ammonia solution.
Apply to the plate 10 µl of each solution. After development, Test solution. Extract a quantity of the powdered tablets
dry the plate in a current of air and examine in ultraviolet light containing 1 mg of Buprenorphine Hydrochloride with three
at 254 nm or expose to iodine vapours. The principal spot in quantities, each of 10 ml, of methanol, filtering each extract
the chromatogram obtained with the test solution corresponds through a sintered-glass filter (porosity No. 4). Evaporate the
to that in the chromatogram obtained with the reference filtrate to dryness and dissolve the residue in 1 ml of methanol.
solution. Reference solution. Dissolve 1 mg of buprenorphine
B. To a volume containing about 5 mg of Buprenorphine hydrochloride RS in 1 ml of methanol.
Hydrochloride in a 125-ml separator, add 1 ml of dilute Apply to the plate 10 µl of each solution. After development,
ammonia solution and shake with three quantities, each of dry the plate in a current of air and examine in ultraviolet light
10 ml, of chloroform. Wash each chloroform extract with the at 254 nm or expose to iodine vapours. The principal spot in
same 10 ml of water and discard the washings. Evaporate the the chromatogram obtained with the test solution corresponds
combined chloroform extracts to dryness on a water-bath and to that in the chromatogram obtained with the reference
dissolve the residue in 50 ml of 0.1 M hydrochloric acid. solution.
When examined in the range 230 to 360 nm (2.4.7) the resulting
B. Shake vigorously a quantity of the powdered tablets
solution shows an absorption maximum only at about 286 nm.
containing 2 mg of Buprenorphine Hydrochloride with 20 ml
of hot water, filter and cool. The filtrate, when examined in the
Tests range 230 nm to 360 nm (2.4.7), shows an absorption maximum
pH (2.4.24). 3.5 to 6.5. at about 286 nm; absorbance at about 286 nm, about 0.33.
Other tests. Complies with the tests stated under Parenteral Tests
Disintegration. The requirement of Disintegration does not
Assay. Measure accurately a volume containing 1.5 mg of apply.
buprenorphine and transfer to a 25-ml volumetric flask. Add
1 ml of 1 M hydrochloric acid, 2 ml of a 2 per cent w/v solution
of sodium nitrite and shake well. Stopper the flask and allow Assay. Weigh and powder 20 tablets. Weigh accurately a
to stand for 15 minutes. Dilute the solution to volume with quantity of the powder containing about 200 mcg of
dilute ammonia solution and measure the absorbance of the buprenorphine and transfer to a 125-ml separator. Add 10 ml
resulting solution at the maximum at about 460 nm (2.4.7), of hot water, shake, add 1 ml of a 10 per cent w/v solution of
using as the blank a solution prepared in the same manner by sodium bicarbonate and shake well. Add 3 ml of a 10 per cent
treating 5 ml of water instead of the preparation under v/v solution of acetic acid, shake, add 3 ml of a 0.2 per cent
examination. w/v solution of metanil yellow and again shake well. Shake
with 100 ml of chloroform for about 5 minutes and allow the
Calculate the content of C29H41NO4 from the absorbance
two layers to separate over a period of 45 minutes. Collect the
obtained by repeating the procedure with 5 ml of a solution
chloroform layer into another 250-ml separator and extract the
containing buprenorphine hydrochloride RS equivalent to
chloroform layer with 50.0 ml of 1 M hydrochloric acid.
0.03 per cent w/v of buprenorphine.
Discard the chloroform layer, centrifuge the red acid layer and
Labelling. The label states the strength in terms of the measure the absorbance at the maximum at about 530 nm (2.4.7),
equivalent amount of buprenorphine in a suitable dose-volume. using 1 M hydrochloric acid as the blank. Calculate the
204
IP 2007 BUSULPHAN TABLETS
content of C29H41NO4 from the absorbance obtained by D. Fuse 0.1 g with 0.1 g of potassium nitrate and 0.25 g of
repeating the procedure with 10.0 ml of a solution containing potassium hydroxide, cool and dissolve the residue in 5 ml of
buprenorphine hydrochloride RS equivalent to 0.002 per cent water. Acidify with dilute hydrochloric acid and add a few
w/v solution of buprenorphine beginning at the words drops of barium chloride solution; a white precipitate is
“transfer to a 125-ml separator....” . produced.
Labelling. The label states the strength in terms of the Tests
equivalent amount of buprenorphine.
Appearance of solution. Dissolve 0.25 g in 20.0 ml of
acetonitrile, dilute to 25 ml with water and examine
immediately. The solution is clear (2.4.1), and not more
Busulphan intensely coloured than reference solution BS6 (2.4.1).
Acidity. Dissolve 0.2 g in 50 ml of warm ethanol previously
O neutralised to methyl red solution and titrate with 0.1 M sodium
O CH3 hydroxide using methyl red solution as indicator; not more
S O
O S than 0.05 ml of 0.1 M sodium hydroxide is required.
H3C O
O
C6H14O6S2 Mol. Wt. 246.3 Loss on drying (2.4.19). Not more than 2.0 per cent, determined
Busulphan is 1,4-butanediol dimethanesulphonate. on 1.0 g by drying in an oven over phosphorus pentoxide at
60° at a pressure of 1.5 to 2.5 kPa.
Busulphan contains not less than 99.0 per cent and not more
than 100.5 per cent of C6H14O6S2, calculated on the dried basis. Assay. Weigh accurately about 0.25 g and shake with 50 ml of
water. Boil under a reflux condenser for 30 minutes and, if
necessary, restore the initial volume with water. Allow to cool
Identification and titrate with 0.1 M sodium hydroxide, using 0.3 ml of dilute
phenolphthalein solution as indicator, until a pink colour is
Test A may be omitted if tests B, C and D are carried out. Tests produced.
A. Determine by infrared absorption spectrophotometry (2.4.6). C6H14O6S2.
Compare the spectrum with that obtained with busulphan RS
or with the reference spectrum of busulphan.
Mobile phase. A mixture of equal volumes of acetone and Busulphan Tablets
toluene. Busulphan Tablets contain not less than 90.0 per cent and not
Test solution. Dissolve 1 g of the substance under examination more than 115.0 per cent of the stated amount of busulphan,
in 100 ml of acetone. C6H14O6S2. The tablets are coated.
Reference solution. A 1 per cent w/v solution of busulphan
Identification
RS in acetone.
Apply to the plate 5 µl of each solution. After development, A. Warm a quantity of the powdered tablets containing 10 mg
dry the plate in a current of hot air, spray with anisaldehyde of Busulphan with 10 ml of acetone, filter and evaporate the
solution and heat at 120°. The principal spot in the filtrate to dryness. Dry the residue at 60° at a pressure not
chromatogram obtained with the test solution corresponds to exceeding 0.7 kPa for 1 hour. The residue complies with the
that in the chromatogram obtained with the reference solution. following test.
C. Heat 0.1 g with 5 ml of 1 M sodium hydroxide until a clear Determine by infrared absorption spectrophotometry (2.4.6).
solution is obtained and allow to cool. To 2 ml of the solution Compare the spectrum with that obtained with busulphan RS
add 0.1 ml of a 3 per cent w/v solution of potassium or with the reference spectrum of busulphan.
permanganate; the purple colour changes to violet, then to B. In the Assay, the principal peak in the chromatogram
blue and finally to green. Filter and add 1 ml of ammoniacal obtained with the test solution corresponds to the peak in the
silver nitrate solution; a precipitate is produced. chromatogram obtained with the reference solution.
205
BUTYLATED HYDROXYANISOLE IP 2007
Tests of the supernatant liquid to 100.0 ml with acetone. To 5.0 ml of

the resulting solution add 5 ml of a 30 per cent solution of
Disintegration (2.5.1). Maximum time, 15 minutes. sodium iodide in acetone, stopper the flask lightly and heat in
Uniformity of content. Comply with the test stated under a water-bath at 50° for 90 minutes. Cool, add 10 ml of the
Tablets. internal standard solution, mix, add 10 ml of water and 20.0 ml
of hexane, shake vigorously for 1 minute and allow to separate.
Determine by gas chromatography (2.4.13).
Use the hexane layer.
Test solution. Add 1 ml of water to one tablet in a 50-ml
Calculate the content of C6H14O6S2 in the tablets.
volumetric flask and place in an ultrasonic bath until
completely dispersed. Add 30 ml of acetone, shake for Storage. Store protected from light.
15 minutes and dilute to 50.0 ml with acetone. Centrifuge and
dilute a quantity of the supernatant liquid with acetone to
produce a solution containing 0.0001 per cent w/v of
Busulphan. To 5.0 ml of the resulting solution add 5 ml of a Butylated Hydroxyanisole
30 per cent w/v solution of sodium iodide in acetone, stopper
BHA
the flask lightly and heat in a water-bath at 50° for 90 minutes.
Cool, add 10 ml of a 0.0001 per cent w/v solution of 1,5-di-
iodopentane (internal standard) in acetone, mix, add 10 ml of OH
water and 20.0 ml of hexane, shake vigorously for 1 minute C(CH3)3
and allow to separate. Use the hexane layer.
Reference solution (a). Add 5 ml of a 30 per cent w/v solution
of sodium iodide in acetone to 5.0 ml of a 0.0001 per cent w/v OCH3
solution of busulphan RS in acetone, stopper the flask lightly
and heat in a water-bath at 50° for 90 minutes. Cool, add 10 ml C11H16O2 Mol. Wt. 180.3
of the internal standard solution, mix, add 10 ml of water and
20.0 ml of hexane, shake vigorously for 1 minute and allow to Butylated Hydroxyanisole is 2-(1,1-dimethylethyl)-4-
separate. Use the hexane layer. methoxyphenol containing not more than 10 per cent of 3-
(1,1-dimethylethyl)-4-methoxyphenol.
Reference solution (b). Prepare in the same manner as
reference solution (a) but using 10 ml of acetone in place of Description. A white or almost white, crystalline powder or
internal standard solution. yellowish-white, waxy solid; odour, aromatic.
Chromatographic system Identification

– a glass column 1.5 m x 4 mm, packed with acid-washed,
diatomaceous support (80 to 100 mesh) coated with A. In the test for Related substances, the principal spot in the
3 per cent w/w of phenyl methyl silicone fluid (50 per chromatogram obtained with test solution (b) corresponds to
cent phenyl), that in the chromatogram obtained with reference solution (a).
– temperature: B. Dissolve about 0.1 g in 10 ml of ethanol (95 per cent), add
column. 140°, 2 ml of a 2.0 per cent w/v solution of sodium tetraborate and
inlet port and detector at 240°, a few crystals of 2,6-dichloroquinone-4-chlorimide; a blue
– electron capture detector, colour is produced (distinction from butylated
– flow rate. 30 ml per minute of the carrier gas. hydroxytoluene).
Calculate the content of C6H14O6S2 in the tablet. C. Dissolve a few crystals in 10 ml of ethanol (95 per cent),
Other tests. Comply with the tests stated under Tablets. add 0.5 ml of a 0.2 per cent w/v solution of potassium
ferricyanide and 0.5 ml of a 0.5 per cent w/v solution of ferric
Assay. Weigh and powder 20 tablets. Determine by gas ammonium sulphate in 0.5 M sulphuric acid; a green to blue
chromatography (2.4.13) as given under the test for Uniformity colour is produced.
of content using the following test solution.
Test solution. Weigh accurately a quantity of the powdered (2.4.17), coating the plate with silica gel G.
tablets containing about 2.5 mg of Busulphan, add 5 ml of
water and place in an ultrasonic bath until completely Mobile phase. Dichloromethane.
dispersed. Add 150 ml of acetone, shake for 15 minutes and Test solution (a). Dissolve 2.5 g of the substance under
dilute to 250.0 ml with acetone. Centrifuge and dilute 10.0 ml examination in 100 ml of dichloromethane.
206
IP 2007 BUTYLATED HYDROXYTOLUENE
Test solution (b). Dilute 1.0 ml of test solution (a) to 10 ml with A. Determine by infrared absorption spectrophotometry (2.4.6).
dichloromethane. Compare the spectrum with that obtained with butylated
Reference solution (a). A 0.25 per cent w/v solution of hydroxytoluene RS.
butylhydroxyanisole RS in dichloromethane. B. When examined in the range 230 nm to 360 nm (2.4.7), a
0.005 per cent w/v solution in ethanol shows an absorption
Reference solution (b). Dilute 1.0 ml of reference solution (a)
to 20 ml with dichloromethane.
between 0.40 and 0.45.
Reference solution (c). Dissolve 50 mg of hydroquinone in
C. Dissolve about 10 mg in 2 ml of ethanol (95 per cent), add
5 ml of ethanol (95 per cent) and dilute to 100 ml with
1 ml of a 0.1 per cent w/v solution of testosterone propionate
dichloromethane. Dilute 1 ml of this solution to 10 ml with
in ethanol (95 per cent) and 2 ml of 2 M sodium hydroxide,
dichloromethane.
heat in a water-bath at 80° for 10 minutes and allow to cool; a
Apply to the plate 5µl of each solution. Allow the mobile blue colour is produced.
phase to rise 10 cm. Dry the plate in air and spray with a
freshly prepared mixture of 10 volumes of potassium D. Dissolve about 0.1 g in 10 ml of ethanol (95 per cent), add
ferricyanide solution, 25 volumes of ferric chloride test 2 ml of a 2.0 per cent w/v solution of sodium tetraborate and
solution and 65 volumes of water. In the chromatogram a few crystals of 2,6-dichloroquinone-4-chlorimide; not more
obtained with test solution (a), any violet-blue spot with an Rf than a faint blue colour is produced (distinction from butylated
value of about 0.35 (due to 3-(1,1-dimethylethyl)-4- hydroxyanisole).
methoxyphenol) is not more intense than the principal spot in E. Dissolve a few crystals in 10 ml of ethanol (95 per cent),
the chromatogram obtained with reference solution (a) (10 per add 0.5 ml of a 0.2 per cent w/v solution of potassium
cent); any spot corresponding to hydroquinone is not more ferricyanide and 0.5 ml of a 0.2 per cent per cent w/v solution
intense than the principal spot in the chromatogram obtained of ferric ammonium sulphate in 0.5 M sulphuric acid; a green
with reference solution (c) (0.2 per cent); any spot, besides to blue colour is produced.
the principal spot and any spots corresponding to 3-(1,1-
dimethylethyl)-4-methoxyphenol and hydroquinone, is not Tests
more intense than the principal spot in the chromatogram
obtained with reference solution (b) (0.5 per cent).
methanol is clear (2.4.1), and not more intensely coloured
Sulphated ash (2.3.18). Not more than 0.05 per cent. than reference solution YS5 or BYS5 (2.4.1).
Storage. Store protected from light. Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel G.
Mobile phase. Dichloromethane.
Butylated Hydroxytoluene Test solution. Dissolve 2 g of the substance under examination
BHT in 100 ml of methanol.
Reference solution. Dilute 1 ml of the test solution to 200 ml
OH with methanol.
(H3C)3C C(CH3)3
dry the plate in air and spray with a freshly prepared mixture of
70 volumes of water, 20 volumes of a 10.5 per cent w/v solution
CH3 of ferric chloride and 10 volumes of potassium ferricyanide
solution. Any secondary spot in the chromatogram obtained
C15H24O Mol. Wt. 220.4 with the test solution is not more intense than the spot in the
Butylated Hydroxytoluene is 2,6-bis(1,1-dimethylethyl)- 4- chromatogram obtained with the reference solution.
methylphenol. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Description. A white to yellowish white, crystalline powder.
Identification
Tests B and C may be omitted if tests A, D and E are carried
out.
207
C
Caffeine ....
Calamine ....
Aqueous Calamine Cream ....
Calamine Lotion ....
Calamine Ointment ....
Calciferol Capsules ....
Calciferol Injection ....
Calciferol Oral Solution ....
Calciferol Tablets ....
Calcium Carbonate ....
Calcium Chloride ....
Calcium Folinate ....
Calcium Folinate Injection ....
Calcium Gluconate ....
Calcium Gluconate Injection ....
Calcium Gluconate Tablets ....
Calcium Lactate ....
Calcium Lactate Tablets ....
Calcium Levulinate ....
Calcium Levulinate Injection ....
Calcium Pantothenate ....
Dibasic Calcium Phosphate ....
Tribasic Calcium Phosphate ....
Calcium Stearate ....
Capreomycin Sulphate ....
Capreomycin Injection ....
Captopril ....
Captopril Tablets ....
Caramel ....
Carbamazepine ....
209
Carbamazepine Tablets ....

Carbenicillin Sodium ....
Carbenicillin Sodium Injection ....
Carbenoxolone Sodium ....
Carbenoxolone Sodium Tablets ....
Carbidopa ....
Carbimazole ....
Carbimazole Tablets ....
Carbomers ....
Carboprost Tromethamine ....
Carboprost Tromethamlne Injection ....
Sodium CarboxymethylCellulose ....
Carnauba Wax ....
Cefaclor ....
Cefaclor capsules ....
Cefaclor Oral Suspension ....
Cefaclor Sustained-release Tablets ....
Cefadroxil ....
Cefadroxil Capsules ....
Cefadroxil Oral Suspension ....
Cefadroxil Tablets ....
Cefazolin Sodium ....
Cefazolin Sodium Injection ....
Cefoperazone Sodium ....
Cefoperazone Injection ....
Cefotaxime Sodium ....
Cefotaxime Sodium Injection ....
Ceftazidime ....
Ceftazidime For Injection ....
Ceftazidime Injection ....
Ceftriaxone Sodium ....
Ceftriaxone Injection ....
210
Cefuroxime Axetil ....

Cefuroxime Axetil Tablets ....
Cefuroxime Sodium ....
Cefuroxime Injection ....
Cellulose Acetate Phthalate ....
Cephalexin ....
Cephalexin Capsules ....
Cephalexin Oral Suspension ....
Cephalexin Tablets ....
Cephaloridine ....
Cephaloridine Injection ....
Cetirizine Hydrochloride ....
Cetirizine Tablets ....
Cetostearyl Alcohol ....
Cetrimide ....
Cetyl Alcohol ....
Activated Charcoal ....
Chlorambucil ....
Chlorambucil Tablets ....
Chloramphenicol ....
Chloramphenicol Capsules ....
Chloramphenicol Eye Drops ....
Chloramphenicol Eye Ointment ....
Chloramphenicol Palmitate ....
Chloramphenicol Oral Suspension ....
Chloramphenicol Sodium Succinate ....
Chloramphenicol Sodium Succinate Injection ....
Chlorbutol ....
Chlorcyclizine Hydrochloride ....
Chlordiazepoxide ....
Chlordiazepoxide Tablets ....
Chlorhexidine Acetate ....
Chlorhaxidine Hydrochloride ....
211
Chlorhexidine Gluconate Solution ....

Chlorobutanol ....
Chlorocresol ....
Chloroform ....
Chloroquine Phosphate ....
Chloroquine Phosphate Injection ....
Chloroquine Phosphate Suspension ....
Chloroquine Phosphate Tablets ....
Chloroquine Sulphate ....
Chloroquine Sulphate Injection ....
Chloroquine Sulphate Tablets ....
Chloroquine Syrup ....
Chloroxylenol ....
Chloroxylenol Solution ....
Chlorpheniramine Maleate ....
Chlorpheniramine Injection ....
Chlorpheniramine Tablets ....
Chlorpromazine Hydrochloride ....
Chlorpromazine Injection ....
Chlorpromazine Tablets ....
Chlorpropamide ....
Chlorpropamide Tablets ....
Chlorthalidone ....
Chlorthalidone Tablets ....
Cholecalciferol ....
Chorionic Gonadotrophin ....
Chorionic Gonadotrophin Injection ....
Ciclesonide ....
Ciclesonide Inhalation ....
Cimetidine ....
Cimetidine Tablets ....
Cinnarizine ....
Cinnarizine Tablets ....
212
Ciprofloxacin ....
Ciprofloxacin Injection ....
Cirpofloxacin Hydrochloride ....
Ciprofloxacin Eye Drops ....
Ciprofloxacin Tablets ....
Cisplatin ....
Cisplatin Injection ....
Citric Acid ....
Citric Acid Monohydrate ....
Clarithromycin ....
Clarithromycin Tablets ....
Clobazam ....
Clobazam Capsules ....
Clofazimine ....
Clofazimine Capsules ....
Clomifene Citrate ....
Clomifene Tablets ....
Clomipramine Hydrochloride ....
Clomipramine Capsules ....
Clonazepam ....
Clonazepam Injection ....
Clonidine Hydrochloride ....
Clonidine Injection ....
Clonidine Tablets ....
Clotrimazole ....
Clotrimazole Cream ....
Clotrimazole Pessaries ....
Clove Oil ....
Cloxacillin Sodium ....
Cloxacillin Capsules ....
Cloxacillin Injection ....
Cloxacillin Syrup ....
Codeine Phosphate ....
213
Codeine Syrup ....

Colchicine ....
Colchicine Tablets ....
Colchicine And Probenecid Tablets ....
Cortisone Acetate ....
Cortisone Injection ....
Cortisone Tablets ....
Absorbent Cotton ....
Cresol ....
Cresol With Soap Solution ....
Croscarmellose Sodium ....
Crospovidone ....
Cyanocobalamin ....
Cyanocobalamin Injection ....
Cyclizine Hydrochloride ....
Cyclizine Tablets ....
Cyclophosphamide ....
Cyclophosphamide Injection ....
Cyclophosphamide Tablets ....
Cyclopropane ....
Cycloserine ....
Cycloserine Capsules ....
Cycloserine Tablets ....
Cyproheptadine Hydrochloride ....
Cyproheptadine Syrup ....
Cyproheptadine Tablets ....
Cytarabine ....
Cytarabine Injection ....
214
IP 2007 CAFFEINE
Mobile phase. A mixture of 40 volumes of 1-butanol,

Caffeine 30 volumes of chloroform, 10 volumes of strong ammonia
solution and 3 volumes of acetone.
O CH3 Test solution. A 2 per cent w/v solution of the substance
H3C N
N under examination in a mixture of 3 volumes of chloroform
and 2 volumes of methanol.
O N N
Reference solution. A 0.01 per cent w/v solution of the
CH3 substance under examination in a mixture of 3 volumes of
chloroform and 2 volumes of methanol.
C8H10N4O2 Mol. Wt. 194.2 (anhydrous) Apply to the plate 10 µl of each solution. After development,
C8H10N4O2,H2O Mol. Wt. 212.2 (monohydrate) dry in air and examine in ultraviolet light at 254 nm. Any
Caffeine is 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione or
its monohydrate.
obtained with the reference solution.
Caffeine contains not less than 98.5 per cent and not more Arsenic (2.3.10). Mix 3.3 g with 3 g of anhydrous sodium
than 101.5 per cent of C8H10N4O2, calculated on the dried basis. carbonate, add 10 ml of bromine solution and mix thoroughly.
Description. Silky white crystals, white glistening needles or Evaporate to dryness on a water-bath, gently ignite and dissolve
a white crystalline powder; odourless; sublimes readily. the cooled residue in 16 ml of brominated hydrochloric acid
and 45 ml of water. Remove the excess of bromine with 2 ml of
Identification stannous chloride solution AsT. The resulting solution
Test A may be omitted if tests B and D are carried out. Tests B,
C and D may be omitted if test A is carried out. Heavy metals (2.3.13). Mix 2.0 gm with 5 ml of 0.1 M
hydrochloric acid and 45 ml of water, warm gently until solution
A. Determine by infrared absorption spectrophotometry (2.4.6), is complete and cool to room temperature. The solution
after drying the substance under examination, at 100° for complies with the limit test for heavy metals, Method A
1 hour. Compare the spectrum with that obtained with caffeine (20 ppm).
RS or with the reference spectrum of caffeine.
B. To 10 mg in a porcelain dish, add 1 ml of hydrochloric acid
Loss on drying (2.4.19). Not more than 0.5 per cent (for the
and 0.1 g of potassium chlorate and evaporate to dryness on
anhydrous form) and 8.5 per cent (for the monohydrate form),
a water-bath. Expose the residue to the vapours of dilute
determined on 1.0 g by drying in an oven at 100° for 1 hour.
ammonia solution; a purple colour is produced which
disappears on addition of a solution of a fixed alkali. Assay. Weigh accurately about 0.18 g and dissolve with
warming in 5 ml of anhydrous glacial acetic acid. For Caffeine
C. To a saturated solution add a few drops of tannic acid Hydrate, use material previously dried at 100° to 105°. Cool,
solution; a white precipitate is produced which is soluble in add 10 ml of acetic anhydride and 20 ml of toluene. Titrate
excess of the reagent. with 0.1 M perchloric acid, determining the end-point
D. To 5 ml of saturated solution add 1.5 ml of 0.05 M iodine, potentiometrically (2.4.25).
the solution remains clear. Add a few drops of dilute 1 ml 0.1 M perchloric acid is equivalent to 0.01942 g of
hydrochloric acid; a brown precipitate is formed which C8H10N4O2.
dissolves on neutralisation with sodium hydroxide solution.
Tests Labelling. The label states whether it is anhydrous or
monohydrate.
(2.4.1) and colourless (2.4.1).
Acidity or alkalinity. Dissolve 0.2 g in 10 ml of boiling water Calamine
and cool. Add 0.1 ml of bromothymol blue solution. The
solution is coloured green or yellow. Titrate with 0.02 M sodium Prepared Calamine
hydroxide to a blue colour; not more than 0.1 ml is required. Calamine is Zinc Oxide with a small proportion of ferric oxide.
Related substances. Determine by thin-layer chromatography Calamine contains not less than 98.0 per cent and not more
(2.4.17), coating the plate with silica gel GF254. than 100.5 per cent of ZnO, calculated on the ignited basis.
215
AQUEOUS CALAMINE CREAM IP 2007
Description. A fine, amorphous, impalpable, pink or reddish- Loss on ignition (2.4.20). Not more than 2.0 per cent,
brown powder. determined on 2.0 g by igniting to constant weight at a
temperature not less than 900°.
Identification
Assay. Weigh accurately about 1.5 g and digest with 50.0 ml of
A. Shake 1 g with 10 ml of dilute hydrochloric acid and filter; 0.5 M sulphuric acid, applying gentle heat until no further
the filtrate gives the reactions of zinc salts (2.3.1). solution occurs. Filter and wash the residue with hot water
B. To 1 g add 10 ml of dilute hydrochloric acid, heat to boiling until the last washing is neutral to litmus paper. To the
and filter. To the filtrate add a few drops of ammonium combined filtrate and washings, add 2.5 g of ammonium
thiocyanate solution; a reddish colour is produced. chloride, cool and titrate with 1 M sodium hydroxide using
methyl orange solution as indicator.
Tests 1 ml of 0.5 M sulphuric acid is equivalent to 0.04068 g of ZnO.
Acid-insoluble substances. Not more than 1 per cent w/w, Storage. Store protected from light and moisture.
determined by the following method. Dissolve 1.0 g in 25 ml of
warm dilute hydrochloric acid. If any insoluble residue
remains, filter, wash with water, dry to constant weight at
105°, cool and weigh.
Aqueous Calamine Cream
Alkaline substances. Digest 1.0 g with 20 ml of warm water, Calamine 40 g
filter and add 2 drops of phenolphthalein solution to the Zinc Oxide 30 g
filtrate. If a red colour is produced, not more than 0.2 ml of Liquid Paraffin 200 g
0.05 M sulphuric acid is required to decolorise it. Self-Emulsifying Glyceryl Monostearate 50 g
Water-soluble dyes. Shake 1.0 g with 10 ml of water and filter; Cetostearyl Alcohol 40 g
the filtrate is colourless. Cetomacrogol 1000 10 g
Ethanol-soluble dyes. Shake 1.0 g with 10 ml of ethanol Phenoxyethanol 5g
(90 per cent) and filter; the filtrate is colourless. Purified Water, freshly boiled and cooled 625 g
Arsenic (2.3.10). Dissolve 1.25 g in 15 ml of brominated Melt together the Cetostearyl Alcohol and Cetomacrogol 1000,
hydrochloric acid AsT, add 45 ml of water and remove the stir until cold and dissolve this mixture and the Self-Emulsifying
excess of bromine with a few drops of stannous chloride Glyceryl Monostearate in the Liquid Paraffin at 60º. Add with
solution AsT. The resulting solution complies with the limit rapid stirring to a solution of the Phenoxyethanol in 450 g of
test for arsenic (8 ppm). the Purified Water at the same temperature and stir until cold.
Triturate the Calamine and the Zinc Oxide with the remainder
Lead. Dissolve 2.0 g in a mixture of 20 ml of water and 5 ml of
of the Purified Water and incorporate in the cream with stirring.
glacial acetic acid, filter and add 0.25 ml of potassium
chromate solution; the solution remains clear for 5 minutes. Aqueous Calamine Cream contains not less than 6.30 per cent
and not more than 7.67 per cent w/w of ZnO.
Calcium. Dissolve 0.5 g in a mixture of 10 ml of water and
2.5 ml of glacial acetic acid by warming on a water-bath, if Identification
necessary and filter. To 0.5 ml of the filtrate, add 15 ml of dilute
ammonia solution and 2 ml of a 2.5 per cent w/v solution of The residue obtained in the Assay is yellow when hot and
ammonium oxalate and allow to stand for 2 minutes; the white when cool.
solution remains clear.
Tests
Soluble barium salts. To the remainder of the filtrate obtained
in the test for Calcium add 2 ml of 1 M sulphuric acid and Other tests. Complies with the tests stated under Creams.
allow to stand for 5 minutes; the solution remains clear. Assay. Weigh accurately about 4 g. Heat carefully, taking care
Chlorides (2.3.12). Dissolve 0.33 g in water with the addition to avoid spurting, until the liquid is completely evaporated
of 1 ml of nitric acid and dilute to 30 ml with water. The and the solid is charred. Ignite the residue to constant weight
resulting solution complies with the limit test for chlorides at a temperature of not less than 900º.
(750 ppm). Storage. Store at a temperature not exceeding 30º. Do not
Sulphates (2.3.17). Dissolve 0.1 g in water with the addition of freeze.
3 ml of 2 M hydrochloric acid, filter and dilute to 60 ml with Labelling. The label states (1) the concentrations of Calamine
water .The resulting solution complies with the limit test for and Zinc Oxide in the preparation; (2) that the preparation is
sulphates (0.6 per cent). intended for external use only; (3) the storage conditions.
216
IP 2007 CALCIFEROL CAPSULES
Calamine Lotion Identification

Calamine 150 g The residue obtained in the Assay is yellow when hot and
Zinc Oxide 50 g white when cool.
Bentonite 30 g Tests
Sodium Citrate 5 g
Other tests. Complies with the tests stated under Ointments.
Liquefied Phenol 5 ml
Assay. Weigh accurately about 1.0 g. Heat gently until the
Glycerin 50 ml
base is completely volatalised or charred. Increase the heat
Purified Water, freshly boiled and until all the carbon is removed and ignite the residue until,
cooled sufficient to produce 1000 ml after further ignition, two successive weighings do not differ
Triturate the Calamine, the Zinc Oxide and the Bentonite with by more than 0.2 per cent of the weight of the residue.
a solution of the Sodium Citrate in about 700 ml of Purified 1g of the residue is equivalent to 0.8034 g of Zn.
Water and add the Liquified Phenol, the Glycerin and sufficient
Purified Water to produce 1000 ml. Storage. Store in well-closed containers, at a temperature not
exceeding 30°.
Identification Labelling. The label states (1) the concentration of Calamine
A. To 2 ml add 2 ml of periodic acid reagent, shake, centrifuge in the preparation; (2) that the preparation is intended for
and add 0.5 ml of the supernatant liquid to 2 ml of ammoniacal external use only; (3) the storage conditions.
silver nitrate solution in a test-tube; a silver mirror is produced
on the walls of the tube.
B. Mix 2 ml with 50 ml of water, centrifuge and decant the Calciferol Capsules
supernatant liquid. Suspend the residue in 20 ml of water, add
1 ml of hydrochloric acid, mix and filter. 5 ml of the filtrate, Calciferol Capsules contain Cholecalciferol or Ergocalciferol
after neutralisation by dropwise addition of 2 M sodium usually as a vegetable oil solution contained in soft gelatin
hydroxide, gives the reactions of zinc salts (2.3.1). capsules.
Calciferol Capsules contain not less than 90.0 per cent and
Tests not more than 125.0 per cent of the stated amount of
cholecalciferol, C27H44O or ergocalciferol, C28H44O.
Microbial contamination (2.2.9). 1 g is free from
Staphylococcus aureus and 10 g is free from Pseudomonas Identification
aeruginosa.
Storage. Store at a temperature not exceeding 30°. Do not Extract a capsule with 5 ml of ethanol-free chloroform, filter
freeze. and to 1 ml of the filtrate add 9 ml of antimony trichloride
solution. The light absorption of the resulting solution shows
Labelling. The label states (1) the concentrations of Calamine an absorption maximum at about 500 nm (2.4.7).
and Zinc Oxide in the preparation; (2) that the preparation is
intended for external use only; (3) that the contents should be Tests
shaken before use; (4) the conditions under which the
Other tests. Comply with the tests stated under Capsules.
preparation should be stored.
Assay. Carry out the following procedure in subdued light.
Empty the contents of 20 capsules, or more if required. Weigh
accurately a quantity of the contents of the capsules containing
Calamine Ointment about 6 mg of Cholecalciferol or Ergocalciferol, add 50 ml of
Calamine 150 g ethanol (95 per cent), 14 ml of glycerin and 20 ml of a 50 per
cent w/v solution of potassium hydroxide. Boil under a reflux
White Soft Paraffin 850 g
condenser for 30 minutes, stirring occasionally, add 110 ml of
Triturate the calamine with part of the White Soft Paraffin water and allow to stand for 10 minutes with occasional
until smooth and gradually incorporate the remainder of the stirring. Cool and add sufficient ethanol (95 per cent) to
White Soft Paraffin. produce 250.0 ml. Shake 5.0 ml of the resulting solution with
Calamine Ointment contains not less than 7.8 per cent and not 25.0 ml of light petroleum (40° to 60°) for 3 minutes and
more than 9.4 per cent w/w of Zn. evaporate 5.0-ml portions, accurately measured, in duplicate,
217
CALCIFEROL ORAL SOLUTION IP 2007
of the extract to dryness in a current of oxygen-free nitrogen. Calculate the per centage w/v of C27H44O or C28H44O taking
Dissolve each residue in 1.0 ml of ethanol-free chloroform, 0.87 g as the value of the weight per ml of the injection.
add rapidly 9.0 ml of antimony trichloride solution and Storage. Store in a single dose container protected from light
measure the absorbance of each solution at the maximum at at a temperature not exceeding 30°.
about 500 nm and 550 nm (2.4.7), 90 to 120 seconds after
adding the reagent. Repeat the operations using 1.0 ml portions, Labelling. The label states (1) that the preparation is for
in duplicate, of a solution containing a known amount of intramuscular use only; (2) the number of Units of antirachitic
cholecalciferol RS or ergocalciferol RS as appropriate, in activity (vitamin D) per ml.
ethanol-free chloroform and beginning at the words “add
rapidly 9.0 ml of.....”. Calculate the content of cholecalciferol
or ergocalciferol, in mg, from the difference between the
absorbances at the maximum at about 500 nm and 550 nm.
Calciferol Oral Solution
Calculate the content of C27H44O or C28H44O in the capsules. Calciferol Oral Drops; Calciferol Solution
Storage. Store protected from light and moisture at a Calciferol Oral Solution is a solution of Cholecalciferol or
temperature not exceeding 30°. Ergocalciferol in a suitable vegetable oil and may be prepared
by warming to 40° a 1 per cent w/v suspension of Cholecalciferol
or Ergocalciferol in a suitable vegetable oil, such as Arachis
Oil, carbon dioxide being bubbled through it to facilitate
solution, and adding a sufficient quantity of the oil to produce
Calciferol Injection a solution containing the stated amount of Cholecalciferol or
Calciferol Injection is a sterile solution of Cholecalciferol or Ergocalciferol.
Ergocalciferol in Ethyl Oleate.
Calciferol Oral Solution contains not less than 85.0 per cent
Calciferol Injection contains not less than 90.0 per cent and and not more than 120.0 per cent of the stated amount of
not more than 110.0 per cent of the stated amount of cholecalciferol, C27H44O or ergocalciferol, C28H44O.
cholecalciferol, C27H44O or ergocalciferol, C28H44O.
Description. A pale yellow, oily liquid; odour, slight but not
Description. A pale yellow, oily liquid. rancid.
To 1 ml of a 0.2 per cent v/v solution of the injection in ethanol- To 1 ml of a 20 per cent v/v solution in ethanol-free chloroform
free chloroform add 9 ml of antimony trichloride solution. add 9 ml of antimony trichloride solution. The light absorption
The resulting solution shows an absorption maximum at about of the resulting solution shows an absorption maximum at
500 nm (2.4.7). about 500 nm (2.4.7).
Tests Tests
Other tests. Comply with the tests stated under Parenteral Other tests. Complies with the tests stated under Oral Liquids.
Preparations (Injections). Assay. Carry out the following procedure in subdued light.
Assay. Carry out the following procedure in subdued light. Weigh accurately about 1.5 g, add 0.1 g of hydroquinone and
Weigh accurately about 0.1 g of the injection and dilute to 25 ml of 0.5 M ethanolic potassium hydroxide, boil under a
50.0 ml with dry 1,2-dichloroethane that has been purified by reflux condenser for 20 minutes, cool and add 50 ml of water.
passing it through a column of silica gel. To 1.0 ml of this Extract with three quantities, each of 30 ml, of ether, wash the
solution add rapidly 9.0 ml of antimony trichloride in 1,2- combined ether extracts with 20 ml of water, then with 20 ml of
dichloroethane solution and measure the absorbance of the 0.5 M potassium hydroxide and finally with successive
resulting solution at the maximum at about 500 nm and 550 nm quantities, each of 20 ml, of water until the washings are no
(2.4.7), 90 to 120 seconds after adding the reagent. Repeat the longer alkaline to phenolphthalein solution. Filter the ether
operation using 1.0 ml of a 0.002 per cent w/v solution of solution through absorbent cotton, wash with two quantities,
cholecalciferol RS or ergocalciferol RS in dry, purified 1,2- each of 10 ml, of ether and evaporate the combined extracts
dichloroethane beginning at the words “add rapidly 9.0 ml and washings to dryness under oxygen-free nitrogen by
of.....”. Calculate the content of C27H44O or C28H44O, in mg, immersion in a water-bath at 50°. Dissolve the residue in about
from the difference between the absorbances at the maximum 10 ml of hexane, transfer to a column (20 cm x 10 mm) packed
at about 500 nm and 550nm. with deactivated alumina and elute continuously with a 15 to
218
IP 2007 CALCIFEROL TABLETS
20 per cent v/v solution of ether in hexane, using a flow rate For tablets containing more than 250 µg, prepare the solution
of 1 to 2 ml per minute and collecting the fraction that contains in the same manner but using 4 ml of water, 12 ml of dimethyl
the calciferol (identified conveniently by testing aliquots of sulphoxide and 100 ml of hexane.
successive 10 ml fractions with antimony trichloride solution). Reference solution (a). A 0.001 per cent w/v solution of
Evaporate the solvent under oxygen-free nitrogen at a cholecalciferol RS or ergocalciferol RS, as appropriate.
temperature not exceeding 50° and dissolve the residue in
5.0 ml of ethanol-free chloroform. Using duplicate 1.0 ml Reference solution (b). Dissolve 50.0 mg of cholecalciferol
portions of this solution add rapidly 9.0 ml of antimony RS or ergocalciferol RS as appropriate in 10 ml of toluene
trichloride solution and measure the absorbance of each without heating and dilute with the mobile phase to 100.0 ml;
solution at the maximum at about 500 nm and 550 nm, 90 to 120 dilute 5.0 ml of this solution to 50.0 ml with the mobile phase.
seconds after adding the reagent (2.4.7). Repeat the operation Reflux 5.0 ml of this solution, under nitrogen, using a water-
using duplicate 1.0-ml portions of a solution containing a bath, for 60 minutes to obtain a solution of cholecalciferol,
known amount of cholecalciferol RS or ergocalciferol RS in precholecalciferol and trans-cholecalciferol. Cool and dilute
ethanol-free chloroform and beginning at the words “add the refluxed solution to 50.0 ml with the mobile phase.
rapidly 9.0 ml of antimony trichloride solution.....”. Calculate Chromatographic system
the content of C27H44O or C28H44O, in mg, from the difference – a stainless steel column 25 cm × 4.6 mm, packed with
between the absorbances at the maximum at about 500 nm and porous silica or ceramic microparticles (3 to 10 µm) (such
550 nm. as Nucleosil 50-S 5 µm),
Determine the weight per ml of the oral solution (2.4.29) and – mobile phase: a mixture of 997 volumes of hexane and
calculate the content of C27H44O or C28H44O, weight in volume. 3 volumes of 1-pentanol,
Storage. Store protected from light and moisture at a
temperature not exceeding 30°.
– a 10 or 20 µl loop injector.
Labelling. The label states the number of Units of antirachitic Inject a suitable volume of reference solution (b). Adjust the
activity (vitamin D) per ml. sensitivity so that the height of the peak due to cholecalciferol
is more than 50 per cent of the full-scale deflection. Record the
chromatograms after five more injections. The approximate
Calciferol Tablets relative retention times calculated with reference to
cholecalciferol are 0.4 for precholecalciferol and 0.5 for trans-
Calciferol Tablets contain Cholecalciferol or Ergocalciferol cholecalciferol. The resolution between precholecalciferol and
Calciferol Tablets contain not less than 90.0 per cent and not trans-cholecalciferol should be not less than 1.0; if necessary
more than 125.0 per cent of the stated amount of cholecalciferol, adjust the proportions of the constituents and flow rate of the
C27H44O or ergocalciferol, C28H44O. mobile phase to obtain the required resolution.
Inject reference solution (a). Adjust the sensitivity so that the
Identification height of the peak due to cholecalciferol or ergocalciferol is
Powder a tablet, extract with 5 ml of ethanol-free chloroform, more than 50 per cent of the full-scale deflection.
filter and to 1 ml of the filtrate add 9 ml of antimony trichloride Inject alternately the test solution and reference solution (a).
solution; a brownish-red colour is produced.
Calculate the content of cholecalciferol, C 27 H 44O, or
Tests ergocalciferol, C28H44O in the tablet.
Tablets. Assay. Carry out the following procedure as rapidly as
possible in subdued light and protected from air.
Weigh and powder 20 or more tablets as required. Weigh
Carry out the following procedure as rapidly as possible in accurately a quantity of the powder containing about 6 mg of
subdued light and protected from air. Ergocalciferol or Cholecalciferol, add 50 ml of ethanol (95 per
Test solution. For tablets containing less than 250 µg, add 2 ml cent), 14 ml of glycerin and 20 ml of a 50 per cent w/v solution
of water to one tablet in an amber-coloured flask and disperse of potassium hydroxide. Boil under a reflux condenser for
with the aid of ultrasound. Add 6 ml of dimethyl sulphoxide, 30 minutes, stirring occasionally, add 110 ml of water and
mix, extract with 25 ml of hexane by shaking for 30 minutes, allow to stand for 10 minutes with occasional stirring. Cool
centrifuge the hexane layer and use the clear supernatant liquid. and add sufficient ethanol (95 per cent) to produce 250.0 ml.
219
CALCIUM CARBONATE IP 2007
Shake 5.0 ml of the resulting solution with 25.0 ml of light Heavy metals (2.3.13). To 1.0 g add 5 ml of water, and 8 ml of
petroleum (40° to 60°) for 3 minutes and evaporate 5.0-ml dilute hydrochloric acid, the latter being added slowly, shake
portions, accurately measured, in duplicate, of the extract to and evaporate to dryness on a water-bath. Dissolve the residue
dryness in a current of oxygen-free nitrogen. Dissolve each in 20 ml of water, filter, add to the filtrate 3 ml of dilute acetic
residue in 1.0 ml of ethanol-free chloroform, add rapidly 9.0 ml acid and water to make 25 ml. The solution complies with the
of antimony trichloride solution and measure the absorbance limit test for heavy metals, Method A (20 ppm).
of each solution at the maximum at about 500 nm and 550 nm Barium. Dissolve 0.6 g in 10 ml of 2 M acetic acid by boiling,
(2.4.7), 90 to 120 seconds after adding the reagent. Repeat the cool and add 10 ml of calcium sulphate solution; the solution
operations using 1.0-ml portions, in duplicate, of a solution remains clear for not less than 15 minutes.
containing a known amount of cholecalciferol RS or
ergocalciferol RS as appropriate, in ethanol-free chloroform Iron (2.3.14). Dissolve 0.2 g in 5 ml water and 0.5 ml of iron-
and beginning at the words “add rapidly 9.0 ml of.....”. free hydrochloric acid, boil and dilute to 40 ml with water, the
Calculate the content of cholecalciferol or ergocalciferol, in solution complies with the limit test for iron (200 ppm).
mg, from the difference between the absorbances at the Magnesium and alkali metals. Dissolve 1.0 g in 10 ml of dilute
maximum at about 500 nm and 550 nm. hydrochloric acid, neutralise the solution by adding dilute
Storage. Store protected from light and moisture, at a ammonia solution, heat the solution to boiling and add 50 ml
temperature not exceeding 30°. of hot ammonium oxalate solution. Cool, dilute to 100 ml with
water and filter. To 50 ml of the filtrate add 1.5 ml of dilute
sulphuric acid, evaporate to dryness on a water-bath, heat
the residue to redness, allow to cool and weigh. The residue
weighs not more than 5 mg (1.0 per cent).
Calcium Carbonate
Chlorides (2.3.12). 1.0 g dissolved in water by the addition of
Precipitated Chalk 3 ml of nitric acid complies with the limit test for chlorides
CaCO3 Mol. Wt. 100.1 (250 ppm).
Calcium Carbonate contains not less than 98.0 per cent and Sulphates (2.3.17). Suspend 50.0 mg in 5 ml of water and add
not more than 100.5 per cent of CaCO3, calculated on the dried dropwise sufficient dilute hydrochloric acid to effect solution.
basis. Add 2 ml of dilute hydrochloric acid; the resulting solution
complies with the limit test for sulphates (0.3 per cent).
Description. A fine, white, microcrystalline powder.
Identification on 1.0 g by drying in an oven at 200°.
A. Dissolve 5.0 g in 80 ml of 2 M acetic acid. When dilute hydrochloric acid and 10 ml of water. Boil for
effervescence ceases, boil the solution for 2 minutes, allow to 10 minutes, cool, dilute to 50 ml with water. Titrate with
cool, dilute to 100 ml with 2 M acetic acid and filter, if 0.05 M disodium edetate to within a few ml of the expected
necessary, through a sintered-glass filter reserving any residue end-point, add 8 ml of sodium hydroxide solution and 0.1 g of
for the test for Substances insoluble in acetic acid; 0.2 ml of calcon mixture and continue the titration until the colour of
the filtrate (solution A) gives reactions A and B of calcium the solution changes from pink to a full blue colour.
salts (2.3.1).
1 ml of 0.05 M disodium edetate is equivalent to 0.005004 g of
B. Gives reaction A of carbonates (2.3.1). CaCO3.
Tests
Substances insoluble in acetic acid. Wash any residue
obtained in Identification test A with four quantities, each of
Calcium Chloride
5 ml, of hot water and dry at 100° for 1 hour; the residue Calcium Chloride Dihydrate
weighs not more than 10 mg (0.2 per cent).
CaCl2,2H2O Mol. Wt. 147.1
Arsenic (2.3.10). Dissolve 2.5 g in 15 ml of brominated
Calcium Chloride contains not less than 97.0 per cent and not
hydrochloric acid and 45 ml of water and remove the excess
more than 103.0 per cent of CaCl2,2H2O.
of bromine with a few drops of stannous chloride solution
AsT. The resulting solution complies with the limit test for Description. A white, crystalline powder or fragments or
arsenic (4 ppm). granules; odourless; hygroscopic.
220
IP 2007 CALCIUM FOLINATE
Identification 1 ml of 0.05 M disodium edetate is equivalent to 0.007351 g of

CaCl2,2H2O.
A. Gives reactions A and B of calcium salts (2.3.1).
B. A 10 per cent w/v solution in carbon dioxide-free water
prepared from distilled water (solution A) gives reaction A of
chlorides (2.3.1).
Tests Calcium Folinate

Appearance of solution. Solution A is clear (2.4.1) and not Leucovorin Calcium
more intensely coloured than reference solution YS6 (2.4.1).
Acidity or alkalinity. To 10 ml of a freshly prepared 10 per H H
cent w/v solution add 2 drops of phenolphthalein solution. H2N N N
H
Titrate with 0.01 M hydrochloric acid or 0.01 M sodium N N
++
hydroxide; not more than 0.2 ml is required. Ca N H
O CHO N COO
hydrochloric acid and 45 ml of water and remove the excess O COO 2
AsT. The resulting solution complies with the limit test for
C20H21CaN7O7 Mol. Wt. 511.5
arsenic (3 ppm).
Calcium Folinate is calcium N-[4-(2-amino-5-formyl-1,4,5,6,7,8-
Aluminium and phosphate. To 10 ml of a 5.0 per cent w/v
hexahydro-4-oxo-6-pteridinyl)methylaminobenzoyl]- L -
solution, add 2 drops of dilute hydrochloric acid and 1 drop
glutamate.
of phenolphthalein solution. Add ammonium chloride-
ammonium hydroxide solution dropwise until the solution is Calcium Folinate contains not less than 95.0 per cent and not
faintly pink, add a few drops in excess and heat the liquid to more than 105.0 per cent of C20H21CaN7O7, calculated on the
boiling; no turbidity or precipitate is produced. anhydrous basis.
Barium. To 10 ml of solution A add 1 ml of calcium sulphate Description. A yellowish white or yellow powder; odourless.
solution. After not less than 15 minutes the solution is not
more opalescent than a mixture of 10 ml of solution A and 1 ml Identification
of distilled water. Determine by infrared absorption spectrophotometry (2.4.6).
Heavy metals (2.3.13). 2.0 g complies with the limit test for Compare the spectrum with that obtained with calcium folinate
heavy metals, Method A (10 ppm). RS.
Iron (2.3.14). Dissolve 2.0 g in 0.5 ml of hydrochloric acid and Tests
25 ml of water; the resulting solution complies with the limit
test for iron (20 ppm). Heavy metals (2.3.13). 4.0 g complies with the limit test for
Magnesium and alkali salts. Dissolve 1.0 g in 50 ml of water,
add 0.5 g of ammonium chloride heat the solution to boiling Water (2.3.43). Not more than 17.0 per cent determined on
and add 50 ml of hot ammonium oxalate solution. Cool, dilute 0.5 g.
to 100 ml with water and filter. To 50 ml of the filtrate add 1.5 ml
Assay. Use only freshly deionised water wherever water is
of dilute sulphuric acid, evaporate to dryness on a water-
specified throughout this procedure. Protect the solutions
bath, heat the residue to redness, allow to cool and weigh.
from unnecessary exposure to light and complete the Assay
The residue weighs not more than 5 mg (1.0 per cent).
without prolonged interruption.
Sulphates (2.3.17). 0.5 g dissolved in 15 ml of distilled water Determine by liquid chromatography (2.4.14).
complies with the limit test for sulphates (300 ppm).
Solvent mixture. Add 15 ml of a 25 per cent w/v solution of
tetrabutylammonium hydroxide in methanol to 900 ml of
water. Titrate with 0.05 M disodium edetate to within a few ml
water, adjust the pH to 7.5 ± 0.1 with 0.67 M sodium dihydrogen
of the expected end-point, add 8 ml of sodium hydroxide
phosphate and dilute with water to 1000 ml.
solution and 0.1 g of calcon mixture and continue the titration
until the colour of the solution changes from pink to a full blue Test solution. A 0.02 per cent w/v solution of the substance
colour. under examination in the solvent mixture.
221
CALCIUM FOLINATE INJECTION IP 2007
Reference solution. A solution containing 0.0175 per cent w/v Assay. Use only freshly deionised water wherever water is
each of calcium folinate RS and folic acid RS in the solvent specified throughout this procedure. Protect the solutions
mixture. from unnecessary exposure to light and complete the Assay
Chromatographic system without prolonged interruption.
– a stainless steel column 30 cm x 4 mm, packed with Determine by liquid chromatography (2.4.14).
octadecylsilyl silica gel (3 to 10 µm), Solvent mixture. Add 15 ml of a 25 per cent w/v solution of
– mobile phase: a mixture of 15 ml of a 25 per cent w/v tetrabutylammonium hydroxide in methanol to 900 ml of
solution of tetrabutylammonium hydroxide, 825 ml of water, adjust the pH to 7.5 ± 0.1 with 0.67 M sodium dihydrogen
water and 125 ml of acetonitrile, previously adjusted phosphate and dilute with water to 1000 ml.
to pH 7.5 ± 0.1 with 0.67 M sodium dihydrogen
phosphate, diluted with water to 1000 ml, Test solution. Transfer an accurately measured volume of the
– flow rate. 1 to 2 ml per minute, Injection containing about 9 mg of folinic acid to a 50-ml
– spectrophotometer set at 254 nm, volumetric flask, dilute to volume with a solution prepared by
– a 20 µl loop injector. adding 15 ml of a 25 per cent w/v solution of tetrabutyl-
ammonium hydroxide in methanol to 900 ml of water, adjusting
Inject the reference solution. The relative retention times for the pH to 7.5 ± 0.1 with 0.67 M sodium dihydrogen phosphate
calcium folinate and folic acid are 1.0 and about 1.6 and diluting with water to 1000 ml. Transfer 25.0 ml of this
respectively. The test is not valid unless the relative standard solution into a 60-ml separator, add 25 ml of dichloromethane,
deviation for replicate injections is not more than 3.6 per cent. shake the mixture, allow the layers to separate and discard the
Inject the test solution and reference solution. dichloromethane extract. Repeat the extraction with two more
quantities, each of 25ml, of dichloromethane, discarding the
Calculate the content of C20H21CaN7O7.. dichloromethane extracts. Filter the aqueous layer, discarding
Storage. Store protected from light and moisture. the first 5 ml of the filtrate, and collect the remaining filtrate in
a glass-stoppered conical flask.
Calcium Folinate Injection each of calcium folinate RS and folic acid RS in the solvent
mixture.
Leucovorin Calcium Injection
Calcium Folinate Injection is a sterile solution of Calcium – a stainless steel column 30 cm x 4 mm, packed with
Folinate in Water for Injection. octadecylsilyl silica gel (3 to 10 µm),
Calcium Folinate Injection contains not less than 90.0 per cent – mobile phase: a mixture of 15 ml of a 25 per cent w/v
and not more than 120.0 per cent of the stated amount of solution of tetrabutylammonium hydroxide, 825 ml of
folinic acid, C20H23N7O7. water and 125 ml of acetonitrile, previously adjusted
to pH 7.5 ± 0.1 with 0.67 M sodium dihydrogen
Description. A clear, yellowish solution. phosphate, diluted with water to 1000 ml,
– flow rate. 1 to 2 ml per minute,
Transfer a volume containing about 6 mg of folinic acid to a – a 20 µl loop injector.
glass-stoppered, 50-ml centrifuge tube, add about 40 ml of Inject the reference solution. The relative retention times for
acetone, mix, centrifuge for a few minutes and decant the calcium folinate and folic acid are 1.0 and about
liquid phase. Repeat the washing with an additional 40 ml of 1.6 respectively. The test is not valid unless the relative
acetone. Dry the precipitate obtained with a stream of dry standard deviation for replicate injections is not more than 3.6
nitrogen. The precipitate complies with the following test. per cent.
Determine by infrared absorption spectrophotometry (2.4.6). Inject the test solution and reference solution.
Compare the spectrum with that obtained with calcium folinate Calculate the content of C20H23N7O7 in the injection. 1 mg
RS. calcium folinate is approximately equivalent to 0.93 mg of folinic
acid.
Tests
Storage. Store in single dose containers preferably of type I
pH (2.4.24). 6.5 to 8.5. glass, protected from light.
Other tests. Complies with the tests stated under Parenteral Labelling. The label states the strength in terms of the
Preparations (Injections). equivalent amount of folinic acid.
222
IP 2007 CALCIUM GLUCONATE INJECTION
Calcium Gluconate phenolphthalein solution; no colour is produced. Add 0.3 ml

of 0.01 M sodium hydroxide; a pink colour is produced.
Arsenic (2.3.10). Dissolve 5.0 g in 50 ml of water and 12 ml of
COO stannated hydrochloric acid AsT. The resulting solution
H OH complies with the limit test for arsenic (2 ppm).
HO H
Ca++ ,H2O Heavy metals (2.3.13). 1.0 g dissolved in 4 ml of dilute
H OH hydrochloric acid and sufficient water to produce 25 ml
H OH complies with the limit test for heavy metals, Method A
CH2OH (20 ppm).
Chlorides (2.3.12). 1.0 g complies with the limit test for
C12H22CaO14,H2O Mol. Wt. 448.4 chlorides (250 ppm).
Calcium Gluconate is calcium D-gluconate monohydrate. Sulphates (2.3.17). 1.0 g complies with the limit test for
sulphates (150 ppm).
Calcium Gluconate contains not less than 98.5 per cent and
not more than 102.0 per cent of C12H22CaO14,H2O. Sucrose and reducing sugars. To 10 ml of 5 per cent w/v
solution in hot water add 2 ml of dilute hydrochloric acid
Description. A white, crystalline powder or granules.
and boil for 2 minutes. Cool, add 15 ml of sodium carbonate
Identification solution, allow to stand for 5 minutes and filter. Add 5 ml of
the clear filtrate to 2 ml of potassium cupri-tartrate solution
A. Determine by thin-layer chromatography (2.4.17), coating and boil for 2 minutes; no red precipitate is formed.
Mobile phase. A mixture of 50 volumes of ethanol (95 per warm water; cool, add 5.0 ml of 0.05 M magnesium sulphate
cent), 30 volumes of water, 10 volumes of strong ammonia and 10 ml of strong ammonia solution and titrate with 0.05 M
solution and 10 volumes of ethyl acetate. disodium edetate using mordant black II mixture as indicator.
Test solution. A 2 per cent w/v solution of the substance From the volume of 0.05 M disodium edetate required subtract
under examination in water, heating if necessary, to 60° in a the volume of the magnesium sulphate solution added.
water-bath to effect solution. 1 ml of the remainder of 0.05 M disodium edetate is equivalent
Reference solution. A 2 per cent w/v solution of calcium to 0.02242 g of C12H22CaO14,H2O.
gluconate RS in water, heating if necessary, to 60° in a water-
bath to effect solution.
Calcium Gluconate Injection
dry the plate at 100° for 20 minutes, cool and spray with a 5 per Calcium Gluconate Injection is a sterile solution of Calcium
cent w/v solution of potassium dichromate in a 40 per cent Gluconate in Water for Injections. Not more than 5.0 per
w/w solution of sulphuric acid. After 5 minutes the principal cent of the Calcium Gluconate may be replaced with a
spot in the chromatogram obtained with the test solution suitable calcium salt as a stabilising agent.
corresponds to that in the chromatogram obtained with the
Calcium Gluconate Injection contains a quantity of calcium
reference solution.
B. To 1 ml of a 3 per cent w/v solution add 0.05 ml of ferric 9.4 per cent of the stated amount of calcium gluconate,
chloride test solution; a yellow colour is produced. C12H22O14Ca, H2O.
C. A 2 per cent w/v solution gives reactions A and B of calcium
Identification
salts (2.3.1).
Tests the plate with silica gel G..
Appearance of solution. A 2.0 per cent w/v solution at 60° is Mobile phase. A mixture of 50 volumes of ethanol (95 per
not more intensely coloured than reference solution YS6 cent), 30 volumes of water, 10 volumes of strong ammonia
(2.4.1). On cooling to room temperature the solution is not solution and 10 volumes of ethyl acetate.
more opalescent than opalescence standard OS2 (2.4.1).
Test solution. Dilute a suitable volume of the substance under
Acidity and alkalinity. Dissolve 0.5 g in 20 ml of water, add examination to obtain a solution containing 2 per cent w/v of
0.1 ml of 0.01 M hydrochloric acid and 0.1 ml of Calcium Gluconate.
223
CALCIUM GLUCONATE TABLETS IP 2007
Reference solution. A 2 per cent w/v solution of calcium Mobile phase. A mixture of 50 volumes of ethanol (95 per
gluconate RS in water, heating if necessary, to 60° in a water- cent), 30 volumes of water, 10 volumes of strong ammonia
bath to effect solution. solution and 10 volumes of ethyl acetate.
Apply to the plate 5 µl of each solution. After development, Test solution. A 2 per cent w/v solution of the substance
dry the plate at 100° for 20 minutes, cool and spray with a 5 per under examination in water, heating if necessary, to 60° in a
cent w/v solution of potassium dichromate in a 40 per cent water-bath to effect solution.
w/w solution of sulphuric acid. After 5 minutes the principal
Reference solution. A 2 per cent w/v solution of calcium
gluconate RS in water, heating if necessary, to 60° in a water-
bath to effect solution.
reference solution.
B. To 1 ml add 0.05 ml of ferric chloride test solution; an intense
dry the plate at 100° for 20 minutes, cool and spray with a 5 per
yellow colour is produced.
cent w/v solution of potassium dichromate in a 40 per cent
C. Gives the reactions of calcium salts (2.3.1). w/w solution of sulphuric acid. After 5 minutes the principal
Tests corresponds to that in the chromatogram obtained with the
reference solution.
pH (2.4.24). 6.0 to 8.2.
B. To 1 ml of a 3 per cent w/v solution add 0.05 ml of ferric
chloride test solution; a yellow colour is produced.
Unit per mg of calcium gluconate.
C. A 2 per cent w/v solution gives reactions A and B of calcium
salts (2.3.1).
preparations (Injections).
Assay. To an accurately measured volume equivalent to 0.5 g Tests
of Calcium Gluconate add 50 ml of water; cool, add 5.0 ml of
0.05 M magnesium sulphate and 10 ml of strong ammonia
solution and titrate with 0.05 M disodium edetate using Assay. Weigh and powder 20 tablets. Weigh accurately a
mordant black II mixture as indicator. From the volume of quantity of the powder containing about 0.5 g of Calcium
0.05 M disodium edetate required subtract the volume of the Gluconate and ignite, gently at first, until free from carbon.
magnesium sulphate solution added. Cool, add 10 ml of water and sufficient dilute hydrochloric
acid, dropwise, to effect complete solution of the residue.
1 ml of the remainder of 0.05 M disodium edetate is equivalent
Neutralise with dilute ammonia solution, add 5.0 ml of 0.05 M
to 0.002004 g of Ca.
magnesium sulphate and 10 ml of strong ammonia solution
Labelling. The label states (1) the strength as a percentage and titrate with 0.05 M disodium edetate using mordant black
w/v of calcium gluconate equivalent to the total amount of II mixture as indicator. From the volume of 0.05 M disodium
calcium present; (2) that solutions containing visible solid edetate required subtract the volume of the magnesium
particles must not be used; (3) the percentage of any added sulphate solution added.
stabilising agent.
to 0.02242 g of C12H22O14Ca,H2O.
Calcium Gluconate Tablets

Calcium Gluconate Tablets contain not less than 95.0 per cent Calcium Lactate
calcium gluconate, C12H22O14Ca,H2O.
H OH
++
Identification Ca
H3C COO
2
A warm filtered solution of the powdered tablets equivalent to
a 10 per cent w/v solution of Calcium Gluconate complies with C6H10CaO6,xH2O Mol. Wt. 218.2 (anhydrous)
the following tests. Calcium Lactate is hydrated calcium (RS)-2-hydroxy-
A. Determine by thin-layer chromatography (2.4.17), coating propionate or mixtures of the calcium salts of (R)-, (S)- and
the plate with silica gel G.. (RS)-2-hydroxypropionic acid.
224
IP 2007 CALCIUM LEVULINATE
Calcium Lactate contains not less than 98.0 per cent and not Storage. Store protected from moisture.
more than 102.0 per cent of C6H10CaO6, calculated on the dried
basis.
Description. White granules or powder; odourless or with Calcium Lactate Tablets
slight but not unpleasant odour. The pentahydrate is
Calcium Lactate Tablets contain Calcium Lactate equivalent
somewhat efflorescent.
to not less than 95.0 per cent and not more than 105.0 per cent
Identification of the stated amount of calcium lactate pentahydrate,
C6H10CaO6,5H2O.
A. A solution acidified with sulphuric acid and warmed with
potassium permanganate develops the odour of Identification
acetaldehyde.
A. Extract a quantity of the powdered tablets with water, filter
B. Gives the reactions of calcium salts and of lactates (2.3.1). and acidify the filtrate with sulphuric acid, add potassium
permanganate and warm; the odour of acetaldehyde is
Tests produced.
Acidity or alkalinity. To 10 ml of a 5.0 per cent w/v solution in B. The powdered tablets, when moistened with hydrochloric
carbon dioxide-free water add 0.1 ml of 0.1 M hydrochloric acid and introduced on a platinum wire into the flame of a
acid and 0.1 ml of phenolphthalein solution; no colour is bunsen burner, gives a brick-red colour to the flame.
developed. Add 0.6 ml of 0.1 M sodium hydroxide; a pink
colour is produced. Tests
Arsenic (2.3.10). Dissolve 5.0 g in 50 ml of water and 12 ml of Disintegration (2.5.1). 30 minutes.
stannated hydrochloric acid. The resulting solution complies
with the limit test for arsenic (2 ppm). Other tests. Comply with the tests stated under Tablets.
Heavy metals (2.3.13). 1.0 g dissolved in 2.5 ml of dilute Assay. Weigh and powder 20 tablets. Weigh accurately a
hydrochloric acid and sufficient water to produce 25 ml quantity of the powder containing about 0.3 g of calcium
complies with the limit test for heavy metals, Method A lactate pentahydrate, dissolve as completely as possible in 50
(20 ppm). ml of water, add 5.0 ml of 0.05 M magnesium sulphate and
10 ml of strong ammonia solution and titrate with 0.05 M
Iron (2.3.14). 0.5 g complies with the limit test for Iron disodium edetate using mordant black II mixture as indicator.
(80 ppm). From the volume of 0.05 M disodium edetate required subtract
Chlorides (2.3.12). Dissolve 1.25 g in 10 ml water, add 2 ml of the volume of the magnesium sulphate solution added.
nitric acid and sufficient water to produce 50 ml; the resulting
solution complies with the limit test for chlorides (200 ppm).
to 0.01542 g of C6H10CaO6,5H2O.
Sulphates (2.3.17). Dissolve 0.1 g in 10 ml of water, add 2 ml of
hydrochloric acid and sufficient water to produce 15 ml; the
resulting solution complies with the limit test for sulphates Labelling. The label states the strength in terms of the
(0.15 per cent). equivalent amount of calcium lactate pentahydrate.
Reducing sugars. Dissolve 1 g in 10 ml of water, add 5 ml of
potassium cupri-tartrate solution and boil; not more than a
slight brick-red precipitate is produced. Calcium Levulinate
on 0.5 g by drying in an oven at 120° for 4 hours. O
++ ,2H2O
Assay. Weigh accurately about 0.3 g, dissolve in 50 ml of Ca
water, add 5.0 ml of 0.05 M magnesium sulphate and 10 ml of H3C COO
2
strong ammonia solution and titrate with 0.05 M disodium
edetate using mordant black II mixture as indicator. From the C10H14CaO6,2H2O Mol. Wt. 306.3
volume of 0.05 M disodium edetate required substract the Calcium Levulinate is calcium di(4-oxopentanoate) dihydrate.
volume of the magnesium sulphate solution added.
Calcium Levulinate contains not less than 97.5 per cent and
1 ml of the remainder of 0.05 M disodium edetate is equivalent not more than 100.5 per cent of C10H14CaO6, calculated on the
to 0.01091 g of C6H10CaO6. dried basis.
225
CALCIUM LEVULINATE INJECTION IP 2007
Description. A white, crystalline or amorphous powder; odour, Identification

faint and suggestive of burnt sugar.
A. To a volume of the injection containing 0.5 g Calcium
Identification Levulinate add 5 ml of 1 M sodium hydroxide and filter. To the
filtrate add 5 ml of iodine solution; a precipitate of iodoform is
A. Dissolve 0.5 g in 5 ml of water, add 5 ml of 1 M sodium produced.
hydroxide and filter. To the filtrate add 5 ml of iodine solution;
a precipitate of iodoform is produced. B. To a volume of the injection containing 0.1 g of Calcium
Levulinate add 5 ml of dinitrophenylhydrazine solution and
B. Dissolve 0.1 g in 2 ml of water, add 5 ml of allow the mixture to stand in an ice-bath for 1 hour. Collect the
dinitrophenylhydrazine solution and allow the mixture to stand precipitate on a filter, wash well with cold water and dry at
in an ice-bath for 1 hour. Collect the precipitate on a filter, 105° for 1 hour; the hydrazone so obtained melts between
wash well with cold water and dry at 105° for 1 hour; the 198° and 206° (2.4.21).
hydrazone so obtained melts between 198° and 206° (2.4.21).
C. Gives the reactions of calcium salts (2.3.1).
C. Gives the reactions of calcium salts (2.3.1).
D. Melting range (2.4.21) 119° to 125°. Tests
Tests pH (2.4.24). 7.0 to 8.5.

Units per mg of calcium levulinate.
Arsenic (2.3.10). Dissolve 3.3 g in 50 ml of water and 12 ml of
stannated hydrochloric acid. The resulting solution complies Other tests. Complies with the tests stated under Parenteral
with the limit test for arsenic (3 ppm). Preparations (Injections).
Assay. To a volume containing 0.2 g of Calcium Levulinate
add 50 ml of water, add 5.0 ml of 0.05 M magnesium sulphate
and 10 ml of strong ammonia solution and titrate with 0.05 M
Reducing sugars. Dissolve 0.5 g in 10 ml of water, add 2 ml of disodium edetate using mordant black II mixture as indicator.
3 M hydrochloric acid, boil for about 10 minutes and cool. From the volume of 0.05 M disodium edetate required subtract
Add 5 ml of sodium carbonate solution allow to stand for the volume of the magnesium sulphate solution added.
5 minutes, dilute with water to 20 ml and filter. Add 5 ml of the
clear filtrate to about 2 ml of potassium cupri-tartrate solution
to 0.01532 g of C10H14CaO6,2H2O.
and boil for 1 minute; no red precipitate is formed immediately.
Loss on drying (2.4.19). 10.5 per cent to 12.0 per cent, Storage. Store in single dose containers.
determined on 0.5 g by drying in an oven at 60° at a pressure
Assay. Weigh accurately about 0.2 g, dissolve in 50 ml of
water, add 5.0 ml of 0.05 M magnesium sulphate and 10 ml of Calcium Pantothenate
strong ammonia solution and titrate with 0.05 M disodium
edetate using mordant black II mixture as indicator. From the
volume of 0.05 M disodium edetate required subtract the OH H
volume of the magnesium sulphate solution added. ++ N
Ca HO COO
1 ml of the remainder of 0.05 M disodium edetate is equivalent H3C CH3 O
to 0.01351 g of C10H14CaO6. 2
C18H32CaN2O10 Mol. Wt. 476.5

Calcium Levulinate Injection Calcium Pantothenate is the calcium salt of (R)-3-(2,4-
dihydroxy-3,3-dimethylbutyramido)propionic acid.
Calcium Levulinate Injection is a sterile solution of Calcium
Levulinate in Water for Injections. Calcium Pantothenate contains not less than 98.0 per cent
and not more than 101.0 per cent of C18H32CaN2O10, calculated
Calcium Levulinate Injection contains not less than 95.0 per
on the dried basis.
calcium levulinate, C10H14CaO6,2H2O. Description. A white powder; slightly hygroscopic.
226
IP 2007 DIBASIC CALCIUM PHOSPHATE

C18H32CaN2O10.
A. In the test for β-Alanine, the principal spot in the
chromatogram obtained with test solution (b) corresponds to Storage. Store protected from moisture.
B. Boil 50 mg in 5 ml of 1 M sodium hydroxide for 1 minute,
cool, and add 5 ml of 1 M hydrochloric acid and 2 drops of Dibasic Calcium Phosphate
ferric chloride test solution; a strong yellow colour is
Calcium Hydrogen Phosphate
produced.
CaHPO4 Mol. Wt. 136.1 (anhydrous)
C. To 50 mg in 2 ml of 1 M sodium hydroxide add 0.1 ml of
copper sulphate solution; a blue colour is produced. CaHPO4,2H2O Mol. Wt. 172.1 (dihydrate)
D. Gives reaction A of calcium salts (2.3.1). Dibasic Calcium Phosphate is anhydrous or contains two
molecules of water of hydration.
Tests Dibasic Calcium Phosphate contains not less than 98.0 per
Appearance of solution. A 5.0 per cent w/v solution in carbon cent and not more than 105.0 per cent of CaHPO4 (for anhydrous
dioxide-free water is clear, (2.4.1) and colourless (2.4.1). material) or of CaHPO4,2H2O (for the dihydrate).
pH (2.4.24). 6.8 to 8.0, determined in a 5.0 per cent w/v solution. Description. A white, crystalline powder; odourless.
Specific optical rotation (2.4.22). +25.0° to +27.5°, determined Identification

at 20° in a 5.0 per cent w/v solution.
A. Gives reaction B of calcium salts (2.3.1).
ß-Alanine. Determine by thin-layer chromatography (2.4.17),
coating the plate with silica gel G. B. Dissolve 0.1 g in a mixture of 5 ml of 2 M nitric acid and 5 ml
of water; the solution gives reaction C of phosphates (2.3.1).
Mobile phase. A mixture of 65 volumes of ethanol and
35 volumes of water. Tests
Test solution (a). A 4 per cent w/v solution of the substance Acid-insoluble substances. Heat 5.0 g with a mixture of 40 ml
under examination in water. of water and 10 ml of hydrochloric acid and dilute to 100 ml
Test solution (b). A 0.4 per cent w/v solution of the substance with water. Filter, wash with hot water until the last washing
under examination in water. is free from chloride and dry the residue at 105° for 1 hour
(0.1 per cent).
Reference solution (a). A 0.4 per cent w/v solution of calcium
pantothenate RS in water. Arsenic (2.3.10). Dissolve 1.0 g in 15 ml of brominated
hydrochloric acid, add 45 ml of water and remove the excess
Reference solution (b). A 0.02 per cent w/v solution of of bromine with a few drops of stannous chloride solution
b-alanine in water. AsT. The resulting solution complies with the limit test for
Apply to the plate 5 µl of each solution. Allow the mobile arsenic (10 ppm).
phase to rise 12 cm. Dry the plate in a current of air, spray with Heavy metals (2.3.13). Dissolve 2.5 g in 20 ml of 2 M
ethanolic ninhydrin solution and heat at 110° for 10 minutes. hydrochloric acid, filter if necessary, and add 6 M ammonia
Any spot corresponding to β-alanine in the chromatogram until a precipitate is formed. Add 2 M hydrochloric acid just
obtained with test solution (a) is not more intense than the enough to dissolve the precipitate and dilute to 50 ml with
spot in the chromatogram obtained with reference solution distilled water (solution A). 10 ml of this solution complies
(b). with the limit test for heavy metals, Method A (40 ppm).
Heavy metals (2.3.13). 1.0 g dissolved in 25 ml of water complies Barium. To 10 ml of solution A add 0.5 ml of 1 M sulphuric
with the limit test for heavy metals, Method A (20 ppm). acid, mix and set aside for 15 minutes. The solution is not
Loss on drying (2.4.19). Not more than 3.0 per cent, determined more opalescent than a mixture of 10 ml of solution A and
on 1.0 g by drying in an oven at 105°. 0.5 ml of distilled water treated in the same manner.
Iron (2.3.14). 2.0 ml of solution A diluted to 10 ml with water
complies with the limit test for Iron (400 ppm).
anhydrous glacial acetic acid. Titrate 0.1 M perchloric acid,
determining the end point potentiometrically (2.4.25). Carry Carbonate. Suspend 1 g in 5 ml of water and add 2 ml of
out a blank titration. hydrochloric acid; no effervescence is produced.
227
TRIBASIC CALCIUM PHOSPHATE IP 2007
Chlorides (2.3.12). Dissolve 0.2 g in water by the addition of Identification

2 ml of nitric acid. The solution complies with the limit test for
chlorides (0.125 per cent). A. Gives reaction B of calcium salts (2.3.1).
Sulphates (2.3.17). Dissolve 30.0 mg in 25 ml of water by the B. Dissolve 0.1 g in a mixture of 5 ml of 2 M nitric acid and 5 ml
addition of 2 ml of hydrochloric acid. The solution complies of water; the solution gives reaction C of phosphates (2.3.1).
with the limit test for sulphates (2.3.17) (0.5 per cent).
Tests
Nitrate. To 0.1 g add 10 ml of water, 10 ml of nitrogen-free
sulphuric acid and 1 ml of indigo carmine solution and heat Acid-insoluble substances. Heat 5.0 g with a mixture of 40 ml
to boiling; the blue colour does not disappear. of water and 10 ml of hydrochloric acid and dilute to 100 ml
with water. Filter, wash with hot water until the last washing
Reducing substances. Shake 1.0 g with 5 ml of water and 5 ml is free from chloride and dry the residue at 105° for 1 hour
of 3 M sulphuric acid for 1 minute. Add 0.1 ml of 0.005 M (0.3 per cent).
potassium permanganate and shake for 20 seconds. The slight
pink colour is not less intense than that produced by treating Water-soluble substances. Digest 2.0 g with 100 ml of water
1 g of calcium carbonate in the same manner. for 30 minutes on a water-bath, cool, add sufficient water to
restore the original volume, stir well and filter. Evaporate 50 ml
Proteinous impurities. Heat 0.5 g gently in a dry test-tube; of the filtrate to dryness and dry the residue at 105° to constant
no change in colour is observed and no unpleasant odour is weight (0.5 per cent).
emitted.
Arsenic (2.3.10). Dissolve 2.0 g in a mixture of 15 ml of
Monocalcium and tricalcium phosphates. Dissolve 2.0 g in brominated hydrochloric acid, add 45 ml of water and remove
30.0 ml of 1 M hydrochloric acid, add 20 ml of water and 0.05 the excess of bromine with a few drops of stannous chloride
ml of methyl orange solution and titrate the excess of acid solution AsT. The resulting solution complies with the limit
with 1 M sodium hydroxide. Not less than 14.0 ml and not test for arsenic (5 ppm).
more than 15.5 ml of 1 M hydrochloric acid (for anhydrous
material) and not less than 11.0 ml and not more than 12.5 ml of Heavy metals (2.3.13). Warm 1.0 g with 4 ml of dilute
1 M hydrochloric acid (for the dihydrate) is required. hydrochloric acid, add sufficient water to produce 50 ml and
filter. 25 ml of this solution complies with the limit test for
Loss on ignition (2.4.20). 6.5 to 8.5 per cent (for anhydrous
material) and 24.5 to 26.5 per cent (for the dihydrate),
determined on 1.0 g by igniting at 500°. Iron (2.3.14). Dissolve 0.2 g in a mixture of 5 ml of water and
0.5 ml of iron-free hydrochloric acid with the addition of 1 g
of citric acid. Dilute the solution to 40 ml with water. The
of 5 ml of water and 1 ml of 7 M hydrochloric acid, add 25.0 ml
solution complies with the limit test for iron (200 ppm).
of 0.1 M disodium edetate and dilute to 200 ml with water.
Neutralise with strong ammonia solution, add 10 ml of Carbonate. Suspend 1 g in 10 ml of water and add 2 ml of
ammonia buffer pH 10.0 and 50 mg of mordant black 11 hydrochloric acid; no effervescence is produced.
mixture and titrate the excess of disodium edetate with 0.1 M Chlorides (2.3.12). Dissolve 0.25 g in 25 ml of water by the
zinc sulphate. addition of 1 ml of nitric acid. The solution complies with the
1 ml of 0.1 M disodium edetate is equivalent to 0.01361 g of limit test for chlorides (0.1 per cent ).
CaHPO4 or 0.01721 g of CaHPO4,2H2O. Sulphates (2.3.17). Dissolve 100.0 mg in water with the aid of
3 ml of 1 M hydrochloric acid and dilute to 60 ml with water.
15 ml of the resulting solution complies with the limit test for
Tribasic Calcium Phosphate sulphates (0.6 per cent).
Calcium Hydroxide Phosphate; Calcium Phosphate Proteinous impurities. Heat 0.5 g gently in a dry test-tube;
no change in colour is observed and no unpleasant odour is
Tribasic Calcium Phosphate consists mainly of tricalcium
emitted.
diorthophosphate, Ca 3 (PO 4 ) 2 , together with calcium
phosphates of more acidic or basic character. Loss on ignition (2.4.20). Not more than 8.0 per cent, determined
on 1.0 g by igniting at 800° for 30 minutes.
Tribasic Calcium Phosphate contains not less than 90.0 per
cent and not more than 100.5 per cent of calcium phosphates, Water (2.3.43). Not more than 2.5 per cent, determined on
calculated as Ca3(PO4)2. 1.0 g.
Description. A white, amorphous powder; odourless or almost Assay. Weigh accurately about 1.0 g and dissolve in 10 ml of
odourless. hydrochloric acid by heating on a water-bath, add 50 ml of
228
IP 2007 CAPREOMYCIN SULPHATE
water, cool and dilute to 250.0 ml with water. To 25.0 ml of the carefully to dryness again, swirling near the dry point to avoid
resulting solution add 30.0 ml of 0.05 M disodium edetate, spattering. Repeat the hydrochloric acid treatment, then cool,
10.0 ml of ammonia buffer pH 10.9 and 100 ml of water and and dissolve the residues in about 10 ml of water. To each
titrate the excess of disodium edetate with 0.05 M zinc chloride solution add 1 drop of phenolphthalein solution and add
using mordant black 11 solution as indicator. sodium hydroxide solution until the solutions just turn pink,
1 ml of 0.05 M disodium edetate is equivalent to 0.00517 g of then add 3 M hydrochloric acid until the solutions become
Ca3 (PO4)2. colourless. Add 1 ml of 1 M acetic acid and a small amount of
charcoal to each solution, and filter through filter paper into
50-ml Nessler cylinders. Wash with water, dilute with water to
40 ml, add 1.2 ml of thioacetamide reagent and 2 ml of pH 3.5
Calcium Stearate acetate buffer to each tube, and allow to stand for 5 minutes;
the color of the test solution does not exceed that of the
Octadecanoic acid, calcium salt
control (10 ppm Pb).
Calcium Stearate, is a compound of calcium with a mixture of
Assay. Boil about 1.2 g accurately weighed, with 50 ml of 1 M
solid organic acids obtained from fats and consists chiefly of
sulphuric acid for about 3 hours using a watch glass cover to
variable proportions of calcium stearate and calcium palmitate.
avoid splattering, or until the separated fatty acid layer is
Calcium Stearate contains the equivalent of not less than clear, adding water, if necessary to maintain the original volume.
9.0 per cent and not more than 10.5 per cent of calcium oxide [Note-Stirring may be helpful in obtaining a clear layer and
(CaO). decreasing extraction time.] Cool, filter, and wash the filter
and the flask thoroughly with water until the last washing is
Identification not acid to litmus. Neutralize the filtrate with 1 M sodium
A. Heat 1 g with a mixture of 25 ml of water and 5 ml of hydroxide to litmus. While stirring, preferably with a magnetic
hydrochloric acid; fatty acids are liberated and appear as an stirrer, titrate with 0.05 M disodium edetate as follows. Add
oily layer floating on the surface of the liquid. The water layer about 30 ml from a 50-ml burette, then add 1 ml of 1 M sodium
gives the tests for calcium (2.3.1). hydroxide and 300 mg of hydroxy naphthol blue, and continue
the titration to a blue end-point.
B. Mix 25 g with 200 ml of hot water, add 60 ml of 2 M sulphuric
acid, and heat the mixture, with frequent stirring, until the 1 ml of 0.05 M disodium edetate is equivalent to 0.002804 g of
separated fatty acid layer is clear. Wash the fatty acids with calcium.
boiling water until free from sulphate, collect them in a small
beaker, and warm on a steam bath until the water has separated
and the fatty acids are clear. Allow the acids to cool, pour off Capreomycin Sulphate
the water layer, melt the acids, filter into a dry beaker, and dry
at 105° for 20 minutes; the fatty acids so obtained congeal at
a temperature not below 54° (2.4.10). OH
O H O NH2
H2N N NH2
Tests N N
H O OH
Loss on drying (2.4.19). Not more than 4.0 per cent, determined NH H HN H
, 2H2SO4
on 1.0 g by drying in an oven at 105°. N N NH2
O
Heavy metals (2.3.13).Place 2.5 g in a porcelain dish, place a O H O
NH
500 mg portion in a second dish to provide the control, and to
each add 5 ml of a 1 in 4 solution of magnesium nitrate in N NH
alcohol. Cover the dishes with 7.5-cm short-stem funnels so H
that the stems are straight up. Heat on a hot plate at low heat
for 30 minutes, then heat at medium heat for 30 minutes, and C25H46N14O12S Mol. Wt. 766.8
cool. Remove the funnels, add 2 ml of standard lead solution Capreomycin Sulphate is the disulphate salt of capreomycin,
(20 ppm Pb) to the control, and heat each dish over a suitable a polypeptide mixture produced by certain strains of
burner until most of the carbon is burned off. Cool, add 10 ml Streptomyces capreolus.
of nitric acid, and transfer the solutions into 250 ml beakers.
It has a potency equivalent to not less than 700 µg and not
Add 5 ml of 70 per cent perchloric acid, cautiously evaporate
more than 1050 µg of capreomycin per mg.
to dryness, add 2 ml of hydrochloric acid to the residues, and
wash down the insides of the beakers with water. Evaporate Description. A white or almost white powder.
229
CAPREOMYCIN INJECTION IP 2007
Identification Sterility (2.2.11). Complies with the test for sterility.

A. When examined in the range 230 nm to 350 nm (2.4.7), a Storage. Store protected from moisture.
an absorption maximum only at 268 nm. Absorbance at
268 nm, about 1.2. Capreomycin Injection
B. When examined in the range 230 nm to 350 nm (2.4.7), a Capreomycin Injection is a sterile material consisting of
0.004 per cent w/v solution in 0.1 M sodium hydroxide shows Capreomycin Sulphate with or without auxiliary agents. It is
an absorption maximum only at 287 nm. Absorbance at filled in a sealed container.
287 nm, about 0.8.
C. It gives reaction A of sulphates (2.3.1). sealed container in the requisite amount of sterile Water for
Tests
Appearance of solution. A 10.0 per cent w/v solution in water Clarity of solution and Particulate matter stated under
is clear (2.4.1), when examined immediately after preparation. Parenteral Preparations (Injections).
pH (2.4.24). 4.5 to 7.5, determined in a 3.0 per cent w/v solution. Storage. The constituted solution should be used immediately
Capreomycin I content. Determine by liquid chromatography after preparation but, in any case, within the period
(2.4.14). recommended by the manufacturer.
Test solution. Dissolve 25 mg of the substance under Capreomycin injection contains an amount of Capreomycin
examination in 100 ml of water. Sulphate equivalent to not less than 90.0 per cent and not
Reference solution. A 0.025 per cent w/v solution of more than 115.0 per cent of the stated amount of capreomycin.
capreomycin sulphate RS in water. The contents of the sealed container comply with the
Chromatographic system requirements stated under Parenteral Preparations
– a stainless steel column 15 cm x 4.6 mm, packed with (Powders for Injection) and with the following requirements.
nitrile groups chemically bonded to porous silica Identification
particles (5 µm) (such as Spherisorb CN),
– mobile phase: 55 volumes of the solution prepared by A. When examined in the range 230 nm to 350 nm (2.4.7), a
dissolving 0.5 g of ammonium bisulphate in 1000 ml of solution containing 0.004 per cent w/v of capreomycin in
water, filter and 45 volumes of methanol, 0.1 M hydrochloric acid shows an absorption maximum only
– flow rate. 1.5 ml per minute, at 268 nm. Absorbance at 268 nm, about 1.2.
– spectrophotometer set at 268 nm, B. When examined in the range 230 nm to 350 nm (2.4.7), a
– a 20 µl loop injector. solution containing 0.004 per cent w/v of capreomycin in
Inject the reference solution. The test is not valid unless the 0.1 M sodium hydroxide shows an absorption maximum only
resolution between the two principal peaks is at least 1.5. at 287 nm. Absorbance at 287 nm, about 0.8.
In the chromatogram obtained with the test solution, the sum Tests
of the areas of the two principal peaks, due to capreomycins
1A and 1B, is not less than 90 per cent of the total areas of all Appearance of solution. A 10.0 per cent w/v solution in water
the peaks. is clear (2.4.1), when examined immediately after preparation.
Sulphated ash (2.3.18). Not more than 3.0 per cent. pH (2.4.24). 4.5 to 7.5, determined in a 3.0 per cent w/v solution.
Loss on drying (2.4.19). Not more than 10.0 per cent, determined Capreomycin I content. Determine by liquid chromatography
on 0.1 g by drying in an oven for 4 hours at 100° at a pressure (2.4.14).
not exceeding 0.7 kPa. Test solution. Dissolve a quantity of the injection containing
Assay. Determine by the microbiological assay of antibiotics about 25 mg of capreomycin in 100 ml of water.
(2.2.10). Reference solution. A 0.025 per cent w/v solution of
Caproemycin Suphate intended for use in the manufacture capreomycin sulphate RS in water.
of parenteral preparations complies with the following Chromatographic system
additional requirements. – a stainless steel column 15 cm x 4.6 mm, packed with
Bacterial endotoxins (2.2.3). Not more than 0.35 Endotoxin nitrile groups chemically bonded to porous silica
Unit per mg of capreomycin. particles (5 µm) (such as Spherisorb CN),
230
IP 2007 CAPTOPRIL TABLETS
– mobile phase: a mixture of 55 volumes of a solution Test solution. A 0.4 per cent w/v solution of the substance
prepared by dissolving 0.5 g of ammonium bisulphate under examination in methanol.
in 1000 ml of water, filtered and 45 volumes of methanol, Reference solution. A 0.4 per cent w/v solution of captopril
– flow rate. 1.5 ml per minute, RS in methanol.
– a 20 µl loop injector. Apply to the plate, in the form of 1-cm bands, 50 µl of each
solution. Allow the mobile phase to rise 12 cm. Dry in air and
spray with a freshly prepared mixture of 1 volume of strong
resolution between the two principal peaks is at least 1.5.
ammonia solution and 6 volumes of a 0.04 per cent w/v solution
In the chromatogram obtained with the test solution, the sum of 5,5‘-dithiobis(2-nitrobenzoic acid) in methanol and allow
of the areas of the two principal peaks, due to capreomycins to stand for 5 minutes. The principal band in the chromatogram
1A and 1B, is not less than 90 per cent of the total areas of all obtained with the test solution corresponds to that in the
the peaks. chromatogram obtained with the reference solution.
Loss on drying (2.4.19). Not more than 10.0 per cent, determined C. Melting range (2.4.21) 104° to 110°.
on 0.1 g by drying in an oven at 100°at a pressure not exceeding
0.7 kPa for 4 hours. Tests
Bacterial endotoxins (2.2.3). Not more than 0.35 Endotoxin Specific optical rotation (2.4.22). –125° to –134°, determined
Unit per mg of capreomycin. in a 1.0 per cent w/v solution in ethanol.
Assay. Determine by the microbiological assay of antibiotics Heavy metals (2.3.13). 0.66 g complies with the limit test for
(2.2.10). heavy metals, Method B (30 ppm).
Storage. Store protected from moisture, at a temperature not Sulphated ash (2.3.18). Not more than 0.2 per cent.
exceeding 25°. Loss on drying (2.4.19). Not more than 1.0 per cent, determined
Labelling. The label states the quantity of Capreomycin on 1.0 g by drying in an oven at 60° at a pressure not exceeding
Sulphate in terms of the equivalent amount of capreomycin. 0.7 kPa.
water in a stoppered-flask, add 10 ml of 1.8 M sulphuric acid
Captopril and 1 g of potassium iodide. Titrate with 0.025 M potassium
iodate using 3 ml of starch solution, added towards the end-
O H point, as indicator.
COOH
HS 1ml of 0.025 M potassium iodate is equivalent to 0.03308 g of
H CH3 C9H15NO3S.
C9H15NO3S Mol. Wt. 217.3
Captopril is 1-[(2S)-3-mercapto-2-methylpropionyl]-L-proline.
Captopril contains not less than 97.5 per cent and not more
than 102.0 per cent of C9H15NO3S, calculated on the dried
Captopril Tablets
basis. Captopril Tablets contain not less than 90.0 per cent and not
Description. A white to off-white, crystalline powder; odour, more than 110.0 per cent of the stated amount of captopril,
characteristic, sulphide-like. C9H15NO3S.
A. Determine by infrared absorption spectrophotometry (2.4.6). Determine by thin-layer chromatography (2.4.17), coating the
Compare the spectrum with that obtained with captopril RS plate with silica gel G..
or with the reference spectrum of captopril. Mobile phase. A mixture of 75 volumes of toluene, 25 volumes
B. Determine by thin-layer chromatography (2.4.17), coating of glacial acetic acid and 1 volume of methanol.
the plate with silica gel G. Test solution. Extract a quantity of the powdered tablets
Mobile phase. A mixture of 75 volumes of toluene, 25 volumes containing 100 mg of Captopril with 25 ml of methanol and
of glacial acetic acid and 1 volume of methanol. centrifuge. Use the clear supernatant liquid.
231
CARAMEL IP 2007
Reference solution. A 0.4 per cent w/v solution of captopril Calculate the content of C9H15NO3S in the tablets.
RS in methanol. Storage. Store protected from moisture.
Apply to the plate, in the form of 1-cm bands, 50 µl of each
solution. Allow the mobile phase to rise 12 cm. Dry in air and
spray with a freshly prepared mixture of 1 volume of strong
ammonia solution and 6 volumes of a 0.04 per cent w/v solution
Caramel
of 5,5‘-dithiobis(2-nitrobenzoic acid) in methanol and allow Burnt Sugar
to stand for 5 minutes. The principal band in the chromatogram
Caramel is a concentrated solution of the product obtained
by heating Sucrose or Dextrose until the sweet taste is
destroyed.
Tests Description. A thick, free-flowing, dark brown liquid; odour,
slight and characteristic.
Apparatus. No 2 Identification
To 20 ml of a 5 per cent w/v solution add 0.5 ml of phosphoric
Speed and time. 50 rpm and 20 minutes. acid; no precipitate is produced.
Withdraw a suitable volume of the medium and filter. Measure Tests
the absorbance (2.4.7) of the filtrate, suitably diluted if
necessary, at the maximum at about 212 nm. pH (2.4.24). 3.0 to 5.5, determined in a 10.0 per cent w/v solution.
Calculate the content of C9H15NO3S in the medium from the Weight per ml (2.4.29). Not less than 1.30 g.
absorbance obtained from a solution of known concentration Acid-stability. Dilute 50 ml of a 1 per cent w/v solution to
of captopril RS. 250 ml with water, add 5 ml of hydrochloric acid and heat
D. Not less than 80 per cent of the stated amount of gently to boiling under reflux. Allow to cool and set aside for
C9H15NO3S. 24 hours; the solution remains clear. Repeat the test on the
same test solution but boil for 30 minutes; the solution remains
Other tests. Complies with the tests stated under Tablets.
clear for 24 hours.
Assay. Protect the solutions from exposure to air and use
within 8 hours of preparation.
Iron (2.3.14). Evaporate 0.4 g to dryness, add 0.2 ml of nitric
Test solution. Dissolve a quantity of the finely powdered tablets acid, ignite and dissolve the residue in 1 ml of dilute nitric
containing about 25 mg of Captopril in 25 ml of the mobile acid. The solution complies with the limit test for Iron
phase with the aid of ultrasound for 15 minutes, centrifuge (100 ppm).
and use the clear supernatant liquid.
Microbial contamination (2.2.9). 1 g is free from Escherichia
Reference solution. A 0.1 per cent w/v solution of captopril coli and Salmonellae.
RS in the mobile phase.
– a stainless steel column 25 cm x 4.6 mm, packed with Storage. Store protected from moisture.
octadecylsilyl silica gel (3 to 10 µm),
– mobile phase: a mixture of 55 volumes of methanol and
45 volumes of water containing 0.05 volumes of Carbamazepine
phosphoric acid,
Inject the reference solution. The test is not valid unless the CONH2
than 2.0 per cent. C15H12N2O Mol. Wt. 236.3
Inject alternately the test solution and the reference solution. Carbamazepine is 5H-dibenz[b,f]azepine-5-carboxamide.
232
IP 2007 CARBAMAZEPINE TABLETS
Carbamazepine contains not less than 97.0 per cent and not – a 20 µl loop injector.
more than 103.0 per cent of C15H12N2O, calculated on the dried Inject reference solution (a). The test is not valid unless the
basis. resolution between the peaks due to carbamazepine and 10,11-
Description. A white or yellowish-white, crystalline powder; dihydrocarbamazepine is more than 1.7.
almost odourless; exhibits polymorphism.
Inject test solution (a). Record the chromatograms for 6 times
Identification the retention time of carbamazepine (about 10 minutes). The
areas of any peaks corresponding to 10,11-
A. Determine by infrared absorption spectrophotometry (2.4.6). dihydrocarbamazepine and iminodibenzyl are not more than
Compare the spectrum with that obtained with carbamazepine the areas of the corresponding peaks in the chromatogram
RS. obtained with reference solution (a) (0.1 per cent each). The
area of any other secondary peak is not more than the area of
the peak due to carbamazepine (0.1 per cent) and the sum of all
obtained with test solution (b) corresponds to the peak in the
the secondary peaks is not more than 5 times the area of the
chromatogram obtained with the reference solution (b).
peak due to carbamazepine ( 0.5 per cent). Ignore any peak
Tests with an area less than 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
Acidity or alkalinity. Stir 1.0 g with 20 ml of carbon dioxide- (0.05 per cent).
free water for 15 minutes and filter. Titrate 10 ml of the filtrate
with 0.1 M sodium hydroxide using 0.05 ml of phenolphthalein Heavy metals (2.3.13). 2.0 g complies with the limit test for
solution as indicator; not more than 0.5 ml is required. Add heavy metals, Method B (10 ppm).
0.15 ml of a 0.05 per cent w/v solution of methyl red and titrate Chlorides (2.3.12). 1.5 g complies with the limit test for chlorides
with 0.01 M hydrochloric acid until the colour changes to (165 ppm).
red; not more than 1.0 ml is required.
(2.4.14).
Assay. Determine by liquid chromatography (2.4.14) as given
examination in methanol and dilute to 50 ml with the same
under the test for Related substances using test solution (b)
solvent. Mix with the aid of ultrasound and dilute 10 ml of this
and reference solution (b).
solution to 20 ml with water.
a mixture of equal volumes of methanol and water.
than 2.0 per cent.
Reference solution (a). Dissolve 7.5 mg of carbamazepine
Inject alternately test solution (b) and reference solution (b).
RS, 7.5 mg of 10,11-dihydrocarbamazepine RS and 7.5 mg of
iminodibenzyl in methanol and dilute to 100 ml with the same Calculate the content of C15H12N2O.
solvent. Dilute 1.0 ml of this solution to 50 ml with a mixture of Storage. Store protected from moisture.
equal volumes of methanol and water.
Reference solution (b). Dissolve 0.15 g of carbamazepine RS
in methanol and dilute to 50 ml with the same solvent. Dilute
5 ml of this solution to 50 ml with a mixture of equal volumes of Carbamazepine Tablets
methanol and water.
Carbamazepine Tablets contain not less than 95.0 per cent
Chromatographic system and not more than 105.0 per cent of the stated amount of
– a stainless steel column 25 cm x 4.6 mm, packed with carbamazepine, C15H12N2O.
nitrile groups chemically bonded to porous silica
particles (10 µm), Identification
– mobile phase: a mixture of 3 volumes of tetrahydrofuran,
12 volumes of methanol and 85 volumes of water adding Boil a quantity of the powdered tablets containing 0.2 g of
0.2 ml of formic acid and 0.5 ml of triethylamine to Carbamazepine with 15 ml of acetone, filter the hot solution,
1000 ml of this solution, wash the filtrate with two 5 ml quantities of hot acetone, cool
– flow rate. 1 ml per minute, in ice, evaporate the combined filtrates to dryness. The residue
– spectrophotometer set at 230 nm, complies with the following test
233
CARBENICILLIN SODIUM IP 2007
Determine by infrared absorption spectrophotometry (2.4.6). Assay. Determine by liquid chromatography (2.4.14) as given
Compare the spectrum with that obtained with carbamazepine under the test for Related substances using the following
RS. solutions.
Tests containing about 0.3 g of Carbamazepine with 100.0 ml of
Related substances. Determine by liquid chromatography methanol for 15 minutes. Dilute to 200.0 ml with water, mix,
(2.4.14). filter and further dilute 1 volume of the filtrate to 5 volumes
with methanol (50 per cent).
Reference solution. A 0.03 per cent w/v solution of
containing 0.3 g of Carbamazepine with 100 ml of methanol for
carbamazepine RS in methanol (50 per cent).
15 minutes. Dilute to 200 ml with water, mix and filter.
Reference solution. Dissolve 7.5 mg each of carbamazepine
RS, 10,11-dihydrocarbamazepine and iminodibenzyl in Calculate the content of C15H12N2O in the tablets.
methanol and dilute to 100 ml with the same solvent. Dilute Storage. Store protected from moisture.
1 ml of the resulting solution to 50 ml with methanol (50 per
cent).
– a stainless steel column 25 cm × 4.6 mm, packed with Carbenicillin Sodium
nitrile groups chemically bonded to porous silica
Carbenicillin Disodium
particles (10 µm) (such as Nucleosil 10 CN),
– mobile phase: a mixture of 30 volumes of
tetrahydrofuran, 120 volumes of methanol and H COONa
O
850 volumes of water, adding 0.2 ml of anhydrous formic O N CH3
acid and 0.5 ml of triethylamine to 1000 ml of the
solution, N S CH3
H H H
– flow rate. 2 ml per minute, NaOOC
– a 20 µl loop injector. C17H16N2Na2O6S Mol. Wt. 422.4
Inject the reference solution. The test is not valid unless in Carbenicillin Sodium is the disodium (6R)-6-[(2RS)-2-
the chromatogram obtained the resolution between the peaks carboxylato-2-phenylacetamido]penicillinate.
due to carbamazepine and 10,11-dihydrocarbamazepine is at Carbenicillin Sodium contains the equivalent of not less than
least 1.7. 770 µg of carbenicillin per mg, calculated on the anhydrous
Inject the test solution and continue the chromatography for basis.
6 times the retention time of carbamazepine which is about Description. A white or slightly yellowish powder; odourless;
10 minutes. hygroscopic.
In the chromatogram obtained with the test solution, the areas
Identification
of any peaks corresponding to 10,11-dihydrocarbamazepine
and iminodibenzyl are not greater than the areas of the A. Determine by infrared absorption spectrophotometry (2.4.6).
corresponding peaks in the chromatogram obtained with the Compare the spectrum with that obtained with carbenicillin
reference solution (0.1 per cent). The area of any other sodium RS or with the reference spectrum of carbenicillin.
secondary peak is not greater than the area of the peak due to
B. Heat 0.5 g in a small sealed container on a water-bath for
carbamazepine (0.1 per cent) and the sum of the areas of any 3 minutes, remove the seal, and immediately replace by a cork
such peaks is not greater than 5 times the area of the peak due fitted with a platinum loop carrying a drop of a solution freshly
to carbamazepine (0.5 per cent). Ignore any peak with an area prepared by mixing 1 ml of a 0.5 per cent w/v solution of
less than 0.5 times the area of the peak due to carbamazepine sodium carbonate, 1 ml of phenolphthalein solution and
in the chromatogram obtained with the reference solution 10 ml of water; the reagent is decolourised within 2 minutes.
(0.05 per cent).
C. A 5 per cent w/v solution gives the reactions of sodium
Other tests. Comply with the tests stated under Tablets. salts (2.3.1).
234
IP 2007 CARBENICILLIN SODIUM INJECTION
Tests Description. A white or almost white powder; odourless;

hygroscopic.
Specific optical rotation (2.4.22). +182° to +196°, determined requirements stated under Parenteral Preparations (Powders
at 20° in a 1.0 per cent w/v solution. for Injection) and with the following requirements.
Iodine-absorbing substances. Not more than 8.0 per cent,
calculated on the anhydrous basis, determined by the following Identification
method. Weigh accurately about 0.13 g and dissolve in
sufficient mixed phosphate buffer pH 7.0 to produce 25.0 ml. A. Determine by infrared absorption spectrophotometry (2.4.6).
To 10.0 ml add 10 ml of mixed phosphate buffer pH 4.0 and Compare the spectrum with that obtained with carbenicillin
10.0 ml of 0.01 M iodine and titrate immediately with 0.01 M sodium RS or with the reference spectrum of carbenicillin.
sodium thiosulphate using starch solution, added towards B. Heat 0.5 g in a small sealed container on a water-bath for
the end of the titration, as indicator. Repeat the operation 3 minutes, remove the seal, and immediately replace by a cork
without the substance under examination. The difference fitted with a platinum loop carrying a drop of a solution freshly
between the titration represents the amount of iodine- prepared by mixing 1 ml of a 0.5 per cent w/v solution of
absorbing substances present. sodium carbonate, 1 ml of phenolphthalein solution and
1 ml of 0.01M sodium thiosulphate is equivalent to 0.000489 g 10 ml of water; the reagent is decolourised within 2 minutes.
of iodine-absorbing substances. C. A 5 per cent w/v solution gives the reactions of sodium
Bacterial endotoxins (2.2.3). Not more than 0.05 Endotoxin salts (2.3.1).
Unit per mg of carbenicillin.
Tests
0.15 g. pH (2.4.24). 6.5 to 8.0, determined in a 10.0 per cent w/v solution.
Assay. Determine by the microbiological assay of antibiotics, Specific optical rotation (2.4.22). +182° to +196°, determined
Method A (2.2.10) and express the result in µg of carbenicillin at 20° in a 1.0 per cent w/v solution.
per mg. Iodine-absorbing substances. Not more than 8.0 per cent,
Storage. Store in sterile containers, sealed so as to exclude calculated on the anhydrous basis, determined by the following
micro-organisms, in a refrigerator (2° to 8°). method. Weigh accurately about 0.13 g and dissolve in
sufficient mixed phosphate buffer pH 7.0 to produce 25.0 ml.
To 10.0 ml add 10 ml of mixed phosphate buffer pH 4.0 and
10.0 ml of 0.01 M iodine and titrate immediately with 0.01 M
sodium thiosulphate using starch solution, added towards
Carbenicillin Sodium Injection the end of the titration, as indicator. Repeat the operation
Carbenicillin Injection; Carbenicillin Disodium Injection without the substance under examination. The difference
between the titration represents the amount of iodine-
Carbenicillin Sodium Injection is a sterile material consisting
absorbing substances present.
of Carbenicillin Sodium, with or without auxilliary
substances. It is filled in sealed containers. 1 ml of 0.01M sodium thiosulphate is equivalent to 0.000489 g
of iodine-absorbing substances.
The injection is constituted by dissolving the contents of a
sealed container in the requisite amount of Water for Injections Bacterial endotoxins (2.2.3). Not more than 0.05 Endotoxin
immediately before use. Unit per mg of carbenicillin.
The constituted solution complies with the requirements for Water (2.3.43). Not more than 6.0 per cent, determined on
Clarity of solution and Particulate matter stated under 0.15 g.
Parenteral Preparations (Injections).
Assay. Determine the weight of the contents of 10 containers
Storage. The constituted solution should be used immediately and determine by the microbiological assay of antibiotics,
after preparation but, in any case, within the period Method A (2.2.10) using the mixed contents of the
recommended by the manufacturer. 10 containers.
Carbenicillin Sodium Injection contains the equivalent of not Labelling. The label states the quantity of Carbenicillin Sodium
less than 90.0 per cent and not more than 110.0 per cent of the contained in the sealed container in terms of the equivalent
stated amount of carbenicillin, C17H18N2O6S. amount of carbenicillin.
235
CARBENOXOLONE SODIUM IP 2007
Carbenoxolone Sodium Specific optical rotation (2.4.22). +132° to +140°, determined

in a 1.0 per cent w/v solution in a mixture of equal volumes of
methanol and 0.02 M sodium carbonate.
H3C COONa Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel F254 (such as
precoated Merck silica gel 60 F254 plates).
O
NaOOC Mobile phase. A mixture of 60 volumes of ethyl acetate,
H3C H3C H CH3
20 volumes of methanol, 11 volumes of water and 1 volume of
H strong ammonia solution.
CH3
O O Test solution. A 1.5 per cent w/v solution of the substance
H
H3C CH under examination in methanol.
3
substance under examination in methanol.
C34H48Na2O7 Mol. Wt. 614.7
Carbenoxolone Sodium is disodium 3β-(3- dry the plate in air and examine in ultraviolet light at 254 nm.
carboxylatopropionyloxy)-11-oxo-olean-12-en-30-oate. Spray with a 1.5 per cent w/v solution of vanillin in sulphuric
Carbenoxolone Sodium contains not less than 97.0 per cent acid (60 per cent) and heat at 105° for 10 to 15 minutes. By
and not more than 103.0 per cent of C34H48Na2O7, calculated both methods of visualisation, any secondary spot in the
on the anhydrous basis. chromatogram obtained with the test solution is not more
Description. A white or pale cream powder; hygroscopic;
reference solution.
irritant to nasal membranes.
Identification 0.6 g.
A. Dissolve 0.1 g in 5 ml of water, just acidify with 2 M
water. Add 30 ml of chloroform and 15 ml of a mixture of
hydrochloric acid, stir well and filter. Wash the residue with
10 volumes of 2 M hydrochloric acid and 90 volumes of water,
water until the washings are no longer acidic and dry to
shake and allow to separate. Add the chloroform layer to
constant weight at 105°. The residue complies with the
40 ml of a 20 per cent w/v solution of sodium chloride, shake
following test.
and allow to separate. Repeat the extraction with four
Determine by infrared absorption spectrophotometry (2.4.6). quantities, each of 15 ml, of chloroform, combine the
Compare the spectrum with that obtained with carbenoxolone chloroform extracts and add sufficient chloroform to produce
sodium RS treated in the same manner or with the reference 100.0 ml. Evaporate 25.0 ml, dry the residue at 100° at a pressure
spectrum of carbenoxolone. of 2 kPa and dissolve in 10 ml of dimethylformamide. Titrate
B. When examined in the range 230 nm to 360 nm (2.4.7), a with 0.1 M tetrabutylammonium hydroxide using thymol blue
0.0025 per cent w/v solution in a mixture of equal volumes of solution as indicator. Carry out a blank titration.
methanol and 0.02 M sodium carbonate shows an absorption 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
maximum only at about 256 nm; absorbance at about 256 nm, 0.03073 g of C34H48Na2O7.
about 0.5.
C. Mix 5 mg with 50 mg of resorcinol and 2 ml of sulphuric
acid (80 per cent). Heat at 200° for 10 minutes, cool, pour into Carbenoxolone Tablets
200 ml of water and add sufficient 5 M sodium hydroxide to
make the mixture just alkaline; an intense green fluorescence Carbenoxolone Sodium Tablets
is produced. Carbenoxolone Sodium Tablets contain not less than 95.0 per
D. A 5 per cent w/v solution gives the reactions of sodium cent and not more than 105.0 per cent of the stated amount of
salts (2.3.1). carbenoxolone sodium, C34H48Na2O7.
Identification
Tests
A. Shake a quantity of the powdered tablets containing 0.2 g
pH (2.4.24). 8.0 to 9.2, determined in a 10.0 per cent w/v solution. of Carbenoxolone Sodium with 10 ml of methanol, filter and
236
IP 2007 CARBIDOPA
evaporate to dryness. The residue complies with the following Carbidopa

tests.
1. When examined in the range 230 nm to 360 nm (2.4.7), a COOH
0.0025 per cent w/v solution in a mixture of equal volumes of
H3C NHNH2 ,H2O
methanol and 0.02 M sodium carbonate shows an absorption HO
OH
about 0.5.
2. Mix 5 mg with 50 mg of resorcinol and 2 ml of sulphuric C10H14N2O4,H2O Mol. Wt. 244.3
acid (80 per cent). Heat at 200° for 10 minutes, cool, pour into Carbidopa is (S)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-
200 ml of water and add sufficient 5 M sodium hydroxide to methylpropionic acid monohydrate.
make the mixture just alkaline; an intense green fluorescence
Carbidopa contains not less than 98.5 per cent and not more
is produced.
than 101.0 per cent of C10H14N2O4, calculated with on the dried
B. A 5 per cent w/v solution of the residue obtained in test A basis.
gives the reactions of sodium salts (2.3.1). Description. A white to creamy white powder; odourless or
practically odourless.
Tests
Identification
(2.4.17), coating the plate with silica gel F254 (such as Tests A and C may be omitted if tests B, D and E are carried
precoated Merck silica gel 60 F254 plates). out. Tests B, D and E may be omitted if tests A and C are
carried out.
Mobile phase. A mixture of 60 volumes of ethyl acetate,
20 volumes of methanol, 11 volumes of water and 1 volume of A. Determine by infrared absorption spectrophotometry (2.4.6).
strong ammonia solution. Compare the spectrum with that obtained with carbidopa RS
or with the reference spectrum of carbidopa.
Test solution. Triturate a quantity of the powdered tablets
containing 0.1 g of Carbenoxolone Sodium with 20 ml of B. When examined in the range 230 nm to 360 nm (2.4.7), a
methanol, filter, evaporate the filtrate to low volume and add 0.004 per cent w/v solution in a 1 per cent v/v solution of
sufficient methanol to produce 10 ml. hydrochloric acid in methanol shows an absorption maximum
only at about 282 nm; absorbance at about 282 nm, about 0.52.
Reference solution. Dilute 3 volumes of the test solution to
100 volumes with methanol. C. Complies with the test for Specific optical rotation.
D. Shake vigorously about 5 mg with 10 ml of water for
1 minute and add 0.3 ml of ferric chloride solution; an intense
dry the plate in air and examine in ultraviolet light at 254 nm.
green colour is produced, which quickly becomes reddish
Spray with a 1.5 per cent w/v solution of vanillin in sulphuric
brown.
acid (60 per cent) and heat at 105° for 10 to 15 minutes. By
both methods of visualisation, any secondary spot in the E. Suspend 20 mg in 5 ml of water and add 5 ml of cupri-
chromatogram obtained with the test solution is not more tartaric solution and heat; the colour of the solution changes
intense than the spot in the chromatogram obtained with the to dark brown and a red precipitate is produced.
reference solution.
Tests
Appearance of solution. Dissolve 0.25 g in 25 ml of 1 M
Assay. Weigh and powder 20 tablets. Triturate a quantity of hydrochloric acid. The solution is clear (2.4.1) and not more
the powdered tablets containing about 75 mg of intensely coloured than reference solution BYS6 or BS6 (2.4.1).
Carbenoxolone Sodium with a small volume of methanol, filter
Specific optical rotation (2.4.22). -22.5° to -26.5°, determined
and add sufficient methanol to produce 250.0 ml. To 10.0 ml
in a solution prepared by dissolving 0.25 g in 25 ml of
add 10 ml of 0.02 M sodium carbonate and sufficient of a
aluminium chloride solution.
mixture of equal volumes of methanol and 0.02 M sodium
carbonate to produce 100.0 ml and measure the absorbance Hydrazine. Determine by thin-layer chromatography (2.4.17),
of the resulting solution at the maximum at about 256 nm coating the plate with silanised silica gel G.
(2.4.7). Calculate the content of C34H48Na2O7 taking 199 as the Mobile phase. A mixture of 2 volumes of methanol and
specific absorbance at the maximum at about 256 nm. 1 volume of water.
237
CARBIMAZOLE IP 2007
Test solution. Place 25 g of strongly basic anion exchange Chromatographic system

resin into each of two stoppered conical flasks, add 150 ml of – a stainless steel column 25 cm x 4.6 mm, packed with
carbon dioxide-free water to each flask and allow to stand for octylsilyl silica gel (5 µm),
30 minutes shaking occasionally. Decant the liquid from both – mobile phase: a mixture of 98 volumes of a 1.4 per cent
flasks and repeat the process with further quantities, each of w/v solution of potassium dihydrogen phosphate and
150 ml, of carbon dioxide-free water. Separately transfer the 2 volumes of methanol,
resin portions into two 100-ml measuring cylinders, 3.5 to 4.5 – flow rate. 1 ml per minute,
cm in internal diameter, using 60 ml of carbon dioxide-free – spectrophotometer set at 282 nm,
water for one portion (A) and 20 ml of carbon dioxide-free – a 20 µl loop injector.
water for the other portion (B). Into each cylinder, insert a Inject reference solution (b). The test is not valid unless the
gas-inlet tube, 2 to 3 mm in internal diameter at the end and resolution between the peaks due to methyldopa and carbidopa
reaching almost to the bottom of the cylinder, and pass a rapid is greater than 4.0.
current of nitrogen for chromatography through each mixture
so that homogeneous suspensions are produced. After Inject the test solution and reference solution (a)
30 minutes, without interrupting the gas flow, add 1 ml of a In the chromatogram obtained with the test solution, the areas
solution prepared by dissolving 0.5 g of the substance under of any peaks corresponding to methyldopa and 3-O-
examination in sufficient 2 M hydrochloric acid to produce methylcarbidopa are not greater than the areas of the
2 ml to cylinder A. After 1 minute stop the gas flow to cylinder corresponding peaks in the chromatogram obtained with
A and transfer the contents, through a moistened filter paper, reference solution (a).
into cylinder B. After 1 minute, stop the gas flow to cylinder B
and immediately pour the solution through a moistened filter Heavy metals (2.3.13). 1.0 g complies with the limit test for
paper into a freshly prepared mixture of 1 ml of a 20 per cent heavy metals, Method B (20 ppm).
w/v solution of salicylaldehyde in methanol and 20 ml of Sulphated ash (2.3.18). Not more than 0.1 per cent.
phosphate buffer solution pH 5.5, shake thoroughly for
Loss on drying (2.4.19). 6.9 to 7.9 per cent, determined on
1 minute and heat in a water-bath at 60° for 15 minutes; the
1.0 g by drying in an oven at 105°.
liquid becomes clear. Allow to cool, add 2 ml of toluene, shake
vigorously for 2 minutes and centrifuge. Vigorously shake the Assay. Weigh accurately about 0.15 g and dissolve in 75 ml of
toluene layer with two quantities, each of 20 ml, of a 20 per anhydrous glacial acetic acid with the aid of gentle heat.
cent w/v solution of sodium metabisulphite and then with Titrate with 0.1 M perchloric acid, determining the end-point
two quantities, each of 50 ml, of water and use the toluene potentiometrically (2.4.25). Carry out a blank titration.
layer. 1 ml of 0.1 M perchloric acid is equivalent to 0.02262 g of
Reference solution. Prepare at the same time and in the same C10H14N2O4.
manner but using 1 ml of a 0.002 per cent w/v solution of
hydrazine sulphate in 2 M hydrochloric acid in place of 1 ml
of the solution of the substance under examination.
Any secondary spot in the chromatogram obtained with the
Carbimazole
test solution showing a yellow fluorescence is not more intense
than the corresponding spot in the chromatogram obtained O O CH3
N S
Methyldopa and 3-O-methylcarbidopa. Determine by liquid
chromatography (2.4.14). N
Test solution. Dissolve 0.1 g in sufficient 0.1 M hydrochloric CH3
acid to produce 10 ml.
Reference solution (a). Dissolve 5 mg of methyldopa RS and C7H10N2O2S Mol. Wt. 186.2
5 mg of 3-O-methylcarbidopa RS in sufficient 0.1 M Carbimazole is ethyl 3-methyl-2-thioxo-4-imidazoline-1-
hydrochloric acid to produce 100 ml. carboxylate.
Reference solution (b). Dissolve 5 mg of carbidopa RS and Carbimazole contains not less than 98.0 per cent and not more
5 mg of methyldopa RS in sufficient 0.1 M hydrochloric acid than 102.0 per cent of C7H10N2O2S, calculated on the dried
to produce 10 ml. basis.
238
IP 2007 CARBIMAZOLE TABLETS
Description. A white or creamy-white, crystalline powder; Assay. Weigh accurately about 50 mg and dissolve in sufficient
odour, characteristic. water to produce 500.0 ml. To 10.0 ml of the solution add 10 ml
of 1 M hydrochloric acid and sufficient water to produce
Identification 100.0 ml and measure the absorbance of the resulting solution
at the maximum at about 291 nm (2.4.7). Calculate the content
Test A may be omitted if tests B and C are carried out. Tests B
of C7H10N2O2S taking 557 as the specific absorbance at
291 nm.
Compare the spectrum with that obtained with carbimazole
RS or with the reference spectrum of carbimazole.
B. In the test for Thiamizole and other related substances, the
principal spot in the chromatogram obtained with test solution Carbimazole Tablets
(b) corresponds to that in the chromatogram obtained with Carbimazole Tablets contain not less than 90.0 per cent and
reference solution (b). not more than 110.0 per cent of the stated amount of
C. To a solution prepared by dissolving about 10 mg in a carbimazole, C7H10N2O2S. The tablets may be coated.
mixture of 50 ml of water and 0.05 ml of dilute hydrochloric
acid, add 1 ml of potassium iodobismuthate solution; a red Identification
precipitate is produced. A. Shake a quantity of the powdered tablets containing 50 mg
of Carbimazole with two quantities, each of 5 ml of chloroform.
Tests Combine the chloroform extracts, filter and evaporate the filtrate
Thiamazole and other related substances. Determine by thin- to dryness. Dry the residue at 60° at a pressure not exceeding
layer chromatography (2.4.17), coating the plate with silica 0.7 kPa for 30 minutes The residue complies with the following
gel GF254. test.
Mobile phase. A mixture of 80 volumes of dichloromethane Determine by infrared absorption spectrophotometry (2.4.6).
and 20 volumes of acetone. Compare the spectrum with that obtained with carbimazole
RS or with the reference spectrum of carbimazole.
Test solution (a). 1 per cent w/v solution of the substance
under examination in dichloromethane. B. To a small quantity of the powdered tablets add 1 drop of
dilute potassium iodobismuthate solution; a scarlet colour is
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with produced.
dichloromethane.
Reference solution (a). Dilute 1 ml of test solution (b) to 20 ml Tests
with dichloromethane.
Thiamazole and other related substances. Determine by thin-
Reference solution (b). A 0.1 per cent w/v solution of layer chromatography (2.4.17), coating the plate with silica
carbimazole RS in dichloromethane. gel GF254.
Reference solution (c). Dissolve 5 mg of thiamazole in Mobile phase. A mixture of 80 volumes of dichloromethane
sufficient dichloromethane to produce 100ml. and 20 volumes of acetone.
Apply to the plate 10 µl of each solution After development, Test solution (a). Shake a quantity of the powdered tablets
allow the plate to dry in air for 30 minutes and examine in containing 10 mg of Carbimazole with 2 ml of chloroform for
ultraviolet light at 254 nm. In the chromatogram obtained with 5 minutes and filter.
test solution (a), any spot corresponding to thiamazole is not
Test solution (b). Dilute 1ml of test solution (a) to 10 ml with
dichloromethane.
with reference solution (c) (0.5 per cent); any spot other than
the principal spot and the spot corresponding to thiamazole is Reference solution (a). Dilute 1ml of test solution (b) to 20 ml
not more intense than the spot in the chromatogram obtained with dichloromethane.
with reference solution (a) (0.5 per cent). Reference solution (b). Dissolve 5 mg of thiamazole in
Sulphated ash (2.3.18). Not more than 0.1 per cent. sufficient dichloromethane to produce 100 ml.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Apply to the plate 10 µl of each solution .After development,
on 1.0 g by drying over phosphorus pentoxide at a pressure allow the plate to dry in air for 30 minutes and examine in
not exceeding 0.7 kPa for 24 hours. ultraviolet light at 254 nm. In the chromatogram obtained with
239
CARBOMERS IP 2007
test solution (a), any spot corresponding to thiamazole is not cresol red solution to 10 ml of a 1 per cent w/v dispersion. A
more intense than the spot in the chromatogram obtained yellow colour is produced.
with reference solution (b) (1 per cent); any spot other than E. It complies with the test for viscosity (2.4.28).
the principal spot and the spot corresponding to thiamazole is
not more intense than the spot in the chromatogram obtained Tests
Uniformity of content. Complies with the test stated under Apparent viscosity. The nominal apparent viscosity is in the
Tablets. range 300 mPa s to 115 000 mPa s. For a product with a nominal
apparent viscosity of 20000 mPa s or greater, the apparent
Test solution. Powder one tablet, add 300 ml of water warmed viscosity is 70.0 per cent to 130.0 per cent of the value stated
to a temperature not exceeding 35°, shake for a few minutes on the label; for a product with a nominal apparent viscosity
and add sufficient water to produce 500.0 ml. Mix well, filter, less than 20 000 mPa s, the apparent viscosity is 50.0 per cent
dilute further, if necessary with water. Complete the Assay to 150.0 per cent of the value stated on the label.
beginning at the words “Measure the absorbance....”.
Dry the substance under examination in vacuum at 80° for
1 hour. Carefully add 2.5 g of the previously dried substance
Assay. Weigh and powder 20 tablets. Weigh accurately a under examination to 500 ml of water in a 1000 ml beaker while
quantity of the powder containing about 40 mg of Carbimazole, stirring continuously at 1000 ± 50 rpm, with the stirrer shaft
add 300 ml of water warmed to a temperature not exceeding set at an angle of 60° to one side of the beaker. Add the
35°, shake for a few minutes and add sufficient water to produce previously dried substance over a period of 45 to 90 seconds,
500.0 ml. Mix well and filter. Dilute 5.0 ml of the filtrate to at a uniform rate, ensuring that loose aggregates of powder
50.0 ml with water and mix well. Measure the absorbance of are broken up and continue stirring at 1000 ± 50 rpm for
the resulting solution at the maximum at about 291 nm (2.4.7). 15 minutes. Remove the stirrer, and place the beaker containing
Calculate the content of C7H10N2O2S taking 557 as the specific the dispersion in a water-bath at 25 ± 0.2° for 30 minutes.
absorbance at the maximum at about 291 nm. Insert the stirrer to a depth necessary to ensure that air is not
Storage. Store protected from light and moisture at a drawn into the dispersion, and while stirring at 300 ± 25 rpm,
temperature not exceeding 30°. titrate with a glass-calomel electrode system to pH 7.3 to
7.8 by adding a 18 per cent w/v solution of sodium hydroxide
below the surface, determining the end-point
Carbomers potentiometrically (2.4.25). The total volume of the 18 per cent
w/v solution of sodium hydroxide used is about 6.2 ml. Allow
Carbomers are high molecular mass polymers of acrylic acid 2-3 minutes before the final pH determination. If the final pH
cross-linked with polyalkenyl ethers of sugars or polyalcohols. exceeds 7.8, discard the preparation, and prepare another using
Carbomers contains not less than 56.0 per cent and not more a smaller amount of sodium hydroxide for titration. Return the
than 68.0 per cent of carboxylic acid (-COOH) groups, neutralised preparation to the water-bath at 25° for 1 hour,
calculated on the dried basis. then perform the viscosity determination without delay to
avoid slight viscosity changes that occur 75 minutes after
Description. A white, fluffy powder, hygroscopic.
neutralisation. Determine the viscosity (2.4.28) with a rotating
Identification viscometer with a spindle rotating at 20 rpm, using a spindle
suitable for the expected apparent viscosity.
Free acrylic acid. Determine by liquid chromatography (2.4.14).
Tests B, C, D may be omitted if tests A and E are carried out.
A. Determine by infrared absorption spectrophotometry (2.4.6). Test solution. Dissolve 0.125 g of the substance under
Compare the spectrum with that obtained with carbomers RS. examination in 25 ml of a 2.5 per cent w/v solution of aluminium
potassium sulphate. Heat the suspension at 50° for 20 minutes
B. Adjust a 1 per cent w/v dispersion to about pH 7.5 with 1 M with shaking. Then shake the suspension at room temperature
sodium hydroxide. A highly viscous gel is formed. for 60 minutes. Centrifuge and use the clear supernatant
C. Add 2 ml of a 10 per cent w/v solution of calcium chloride solution.
with continuous stirring to 10 ml of the gel obtained in test B. Reference solution. Dissolve 62.5 mg of acrylic acid RS in
A white precipitate is immediately produced.
100 ml of a 2.5 per cent w/v solution of aluminium potassium
D. Add 0.5 ml of thymol blue solution to 10 ml of a 1 per cent sulphate. Dilute 1.0 ml of this solution to 50.0 ml with 2.5 per
w/v dispersion. An orange colour is produced. Add 0.5 ml of cent w/v solution of aluminium potassium sulphate.
240
IP 2007 CARBOPROST TROMETHAMINE
Chromatographic system – equilibration time 60 minutes,

– a stainless steel column 12 cm x 4.6 mm, packed with – transfer line temperature 90°.
octadecylsilane bonded to porous silica (5 µm), Inject 1 ml of the gaseous phase of the test solution and 1 ml
– mobile phase: A. dissolve 0.136 g in 100 ml of potassium of the gaseous phase of the reference solution; repeat these
dihydrogen phosphate, adjust to pH 2.5 using dilute injections twice more. Maximum relative standard deviation
phosphoric acid, of the differences in area between the analyte peaks obtained
B. equal volumes of a solution of 0.136 g from the 3 replicate pair injections of the reference solution
of potassium dihydrogen phosphate in 100 ml of water and the test solution is 15 per cent. The test is not valid unless
and acetonitrile, the relative standard deviation for replicate injections is not
– flow rate. 1 ml per minute, more than 15 per cent.
below, The mean area of the peak corresponding to benzene in the
– spectrophotometer set at 205 nm, chromatograms obtained with the test solution is not more
– a 20 µl loop injector. than half the mean area of the peak corresponding to benzene
in the chromatograms obtained with the reference solution
Time Mobile Mobile Comments
(2 ppm).
phase A phase B
(in min.) (per cent v/v) (per cent v/v) Heavy metals (2.3.13). 1.0 g complies with the limit test for
0–8 100 → 0 0 isocratic heavy metals, Method C (20 ppm). Use 2 ml of lead standard
solution (10 ppm Pb).
9 – 20 0 → 100 100 linear gradient
Sulphated ash (2.3.18). Not more than 4.0 per cent, determined
21– 30 100 → 0 0 isocratic
on 1.0 g.
The retention time for acrylic acid is about 6.0 minutes. The
on 1.0 g by drying in vacuum at 80° for 60 minutes.
area of the peak in the chromatogram obtained with the test
solution is not more than the area of the corresponding peak Assay. Weigh accurately about 0.12 g, add 50 ml of water
in the chromatogram obtained with the reference solution slowly with stirring and heating at 60° for 15 minutes. Stop
(0.25 per cent). heating, add 150 ml of water and continue stirring for
Benzene. Determine by gas chromatography (2.4.13). 30 minutes. Add 2 g of potassium chloride and titrate with
0.2 M sodium hydroxide determining the end-point
Diluent. Dissolve 0.1 g of benzene in 100 ml of dimethyl potentiometrically (2.4.25).
sulphoxide. Further dilute 1.0 ml of the solution to 100.0 ml
with water. Further dilute 1.0 ml of this solution to 100.0 ml
carboxylic acid (-COOH) groups.
with water.
Test solution. Weigh 50.0 mg of the substance under
examination, add 5.0 ml of water and 1.0 ml of dimethyl Labelling. The label states the nominal apparent viscosity.
sulphoxide.
Reference solution. Weigh 50.0 mg of the substance under
examination, add 4.0 ml of water, 1.0 ml of dimethyl sulphoxide
and 1.0 ml of the diluent. Carboprost Tromethamine
Close the vials with a tight rubber membrane stopper coated
HO NH2
with polytetrafluoroethylene and secure with an aluminium
crimped cap. Shake to obtain a homogeneous dispersion. COOH HO OH
,
Chromatographic system CH3
– a glass column 1.5 m x 4 mm, packed with porous polymer HO
HO H3C OH
beads (80 to 100 mesh),
– temperature :
C21H36O5,C4H11NO3 Mol. Wt. 489.65
column 130°,
inlet port and detector at 240°, Carboprost Tromethamine is a salt of (5Z,13E)-
– flow rate. 30 ml per minute of the carrier gas. (8R,9S,11R,12R,15S)-9,11,15-trihydroxy-15-methylprosta-
Stratic head-space conditions which may be used: 5,13-dienoic acid with 2-amino-2-hydroxymethyl-1,3-
– equilibration temperature 80°, propanediol.
241
CARBOPROST TROMETHAMINE INJECTION IP 2007
Carboprost Tromethamine contains not less than 95.0 per cent (internal standard) in the mobile phase, mix and filter the
and not more than 105.0 per cent of C21H36O5,C4H11NO3, resulting solution through a fine porosity filter.
calculated on the dried basis. Reference solution. Prepare in the same manner but using
Great care should be taken to prevent inhaling particles of caroboprost tromethamine RS in place of the substance under
Carboprost Tromethamine and exposing the skin to it. examination.
Description. A white powder. Chromatographic system
– stainless steel column 30 cm x 4 mm, packed with porous
Identification silica particles (3 to 10 µm),
Determine by infrared absorption spectrophotometry (2.4.6). – mobile phase: a mixture of 7 ml of 1,3-butanediol, 0.5 ml
Compare the spectrum with that obtained with carboprost of water and 992 ml of dichloromethane,
tromethamine RS. Examine the substances as mulls. – flow rate. 1.5 ml per minute,
– spectrophotometer set 254 nm,
Tests – a 10 µl loop injector.
Specific optical rotation. (2.4.22) + 18.0° to + 24.0°, determined Inject the reference solution. The retention times for
in a 1.0 per cent w/v solution in ethanol (95 per cent). guaiphenesin and the 2-naphthacyl ester of carboprost are
about 7 minutes and 11 minutes respectively. The test is not
15R-Epimer and 5-trans isomer. Determine by liquid valid unless the resolution between these two peaks is greater
chromatography (2.4.14). than 4.0 and the relative standard deviation for 4 replicate
Follow the method described under Assay but using 25 µl injections is not more than 2.0 per cent.
loop injector. The usual order of elution is guaiphenesin, the Inject alternately the test solution and the reference solution.
2-naphthacyl ester of 15R-epimer, the 2-naphthacyl ester of
carboprost and the 2-naphthacyl ester of the 5-trans isomer Calculate the content of C21H36O5,C4H11NO3.
with retention times of about 7, 8, 11 and 13 minutes Storage. Store in a refrigerator (2° to 8°).
respectively. Measure the peak areas for the four components
and calculate the contents of the 15R-epimer and 5-trans
isomer. The percentages of I5R-epimer (as tromethamine salt)
and 5-trans isomer are not more than 2.0 per cent and 4.0 per Carboprost Tromethamine Injection
cent respectively.
Carboprost Tromethamine Injection is a sterile solution of
Loss on drying. Not more than 1.0 per cent, determined on Carboprost Tromethamine in Water for Injections. It may
1.0 g by drying in an oven at 50° for 16 hours at a pressure not contain Benzyl alcohol, Sodium Chloride and Tromethamine.
exceeding 0.7 kPa.
Carboprost Tromethamine Injection contains not less than
Assay. Determine by liquid chromatography (2.4.14). 90.0 per cent and not more than 110.0 per cent of the stated
Test Solution. Weigh accurately about 5 mg of the substance amount of carboprost, C21H3605.
under examination, transfer to a stoppered 50-ml centrifuge Description. A colourless solution.
tube. Add 20.0 ml of dichloromethane and 2 ml of citrate buffer
prepared by dissolving 10.5 g of citric acid monohydrate in Identification
about 75 ml of water, adjusting the pH of the solution to 4.0 by
addition of sodium hydroxide solution slowly and diluting to Extract a volume of the injection containing 2.5 mg of
100 ml with water. Shake the stoppered tube for about Carboprost Tromethamine with 1.5 to 2 times its volume of
10 minutes and centrifuge. Transfer 4.0 ml of the lower chloroform. Discard the chloroform Iayer and acidify the
dichloromethane layer to a suitable vial and evaporate the aqueous layer with 3 to 5 drops of hydrochloric acid. Extract
solvent with the aid of a stream of nitrogen. To the dried the acidified solution with an equivalent volume of chloroform.
material add 100 µl of a freshly prepared 2 per cent w/v solution Filter the chloroform layer through a pledget of cotton and
of α-bromo-2'-acetonaphthone in acetonitrile and swirl to concentrate the filtrate to a volume of less than 1 ml. To the
wash down the sides of the vial. Add 50 mI of a freshly prepared resulting solution add 150 mg to 180 mg of potassium bromide
1 per cent v/v solution of diisopropylethylamine in IR and mix well. Dry the potassium bromide mixture in vacuum
acetonitrile, swirl again and place the vial at a temperature of overnight and prepare a disc from the dried mixture.
30° to 35° for not less than 15 minutes. Evaporate the Determine by infrared absorption spectrophotometry (2.4.6).
acetonitrile from the vial with the aid of a stream of nitrogen, Compare the spectrum with that obtained with carboprost
add 2.0 ml of a 0.7 per cent w/v solution of guaiphenesin tromethamine RS treated in the same manr.rost.
242
IP 2007 SODIUM CARBOXYMETHYL CELLULOSE
Tests Calculate the quantity, in µg, of carboprost C21H36O5 per ml of

the injection from the ratios of the peak response of the
pH (2.4.24). 7.0 to 8.0. 2-naphthacyl ester of carboprost and the internal standard
Bacterial endotoxins. Not more than 714.3 Endotoxin Units obtained with the test solution, the ratios of the peak response
per mg of carboprost tromethamine. of the 2-naphthacyl ester of carboprost and the internal
standard obtained with the reference solution and the
concentration, in µg per ml, of carboprost in carboprost
tromethamine RS in the reference solution.
Storage. Store in a refrigerator (2° to 8°).
Test solution. Transfer a volume of the injection containing
500 µg of carboprost to a stoppered 50-ml centrifuge tube. Labelling. The label states the strength in terms of the
Add 20.0 ml of dichloromethane and 1.0 ml of citrate buffer equivalent amount of carboprost in a suitable dose-volume.
prepared by dissolving 10.5 g of citric acid monohydrate in
about 75 ml of water, adjusting the pH of the solution to 4.0 by
addition of sodium hydroxide solution slowly and diluting to
100.0 ml with water. Shake the stoppered tube for about Sodium Carboxymethyl Cellulose
10 minutes and centrifuge. Transfer 8.0 ml of the lower
dichloromethane layer to a suitable vial and evaporate the Carmellose Sodium
solution with the aid of a stream of nitrogen (The residue may Sodium Carboxymethylcellulose is the sodium salt of a
not evaporate to dryness because of the presence of benzyl partially-substituted poly(carboxymethyl) ether of cellulose.
alcohol). Add 100 µI of a freshly prepared 2 per cent w/v
solution of α-bromo-2'-acetonaphthone in acetonitrile and Sodium Carboxymethylcellulose contains not less than
swirl to wash down the sides of the vial. Add 50 µl of a freshly 6.5 per cent and not more than 10.8 per cent of sodium, Na,
prepared 1 per cent v/v solution of diisopropylethylamine in calculated on the dried basis.
acetonitrile, swirl again and place the vial at a temperature of Description. A white or almost white, granular powder;
30° to 35° for not less than 15 minutes. Evaporate the odourless or almost odourless; hygroscopic.
acetonitrile from the vial with the aid of a stream of nitrogen,
add 1.0 ml of a 0.3 per cent w/v solution of guaiphenesin Identification
(internal standard) in the mobile phase, mix and filter the
resulting solution through a fine porosity filter. A. Sprinkle a quantity containing 1.0 g of the dried substance
Reference solution. Prepare an aqueous solution containing on to 90 ml of carbon dioxide-free water at 40° to 50°, stir
about 0.332 mg of carboprost tromethamine RS and 9 mg of vigorously until a colloidal solution is produced, cool and
benzyl alcohol per ml. Transfer 2.0 ml of the resulting solution dilute to 100 ml with carbon dioxide-free water (solution A).
to a stoppered 50-ml centrifuge tube and proceed as given To 10 ml of solution A add 1 ml of copper sulphate solution; a
under the test solution beginning at the words “Add 20.0 ml blue, cotton-like precipitate is produced.
of dichloromethane.... “. B. Boil 5 ml of solution A for a few minutes; no precipitate is
Chromatographic system produced.
– a stainless steel column 30 cm x 4 mm, packed with porous C. Solution A gives the reactions of sodium salts (2.3.1).
silica particles (3 to 10 µm),
– mobile phase: a mixture of 7 ml of 1,3-butanediol, Tests
0.5 ml of water and 992 ml of dichloromethane,
– flow rate. 1.5 ml per minute, Appearance of solution. Solution A is not more opalescent
– spectrophotometer set 254 nm, than opalescence standard OS4 (2.4.1), and not more intensely
– a 10 µl loop injector. coloured than reference solution YS6 (2.4.1).
Inject the reference solution. The retention times for pH (2.4.24). 6.0 to 8.0, determined in solution A.
guaiphenesin and the 2-naphthacyl ester of carboprost are
Apparent viscosity. 75 to 140 per cent of the declared value,
about 7 minutes and 11 minutes respectively. The test is not
determined by the following method. To 50 ml of water heated
valid unless the resolution between these two peaks is greater
to 90° add, with stirring, a quantity containing 2 g of the dried
than 4.0 and the relative standard deviation for replicate
substance under examination or, for a product of low viscosity,
use the quantity required to give the concentration on the
Inject alternately the test solution and the reference solution. label. Allow to cool, dilute to 100 ml with water and continue
243
CARNAUBA WAX IP 2007
stirring until solution is complete. Determine the viscosity by Carnauba Wax

Method C (2.4.28), at 20° using a shear rate of 10 s–1. If
necessary, use rates slightly below and slightly above 10 s-1 Carnauba Wax is obtained from the leaves of Copernicia
and interpolate. cerifera Mart. (Fam. Palmae) after purification to remove
foreign matter.
Arsenic (2.3.10). Place 5.0 g in a dry Kjeldahl flask, add 20 ml
of nitric acid, and warm cautiously until the reaction Description. A pale yellow to light brown coarse powder, flakes
commences. Allow the reaction to subside without further or lumps of hard brittle wax; odour, characteristic and free
heating, then add a mixture of 20 ml of nitric acid and 5 ml of from rancidity.
sulphuric acid and heat until brown fumes cease to be
Identification
evolved. Add 0.5 ml of perchloric acid (60 per cent), heat
until white fumes appear, and if the liquid is still dark add Determine by thin-layer chromatography (2.4.17), coating the
further small quantities of nitric acid and heat until the liquid plate with silica gel G.
becomes pale yellow. Heat again until the white fumes appear
Mobile phase. A mixture of 98 volumes of chloroform and
and continue heating for a further 15 minutes. Add 0.5 ml of
perchloric acid (60 per cent) and continue heating for a few
minutes. Allow the solution to cool add 10 ml of water, and Test solution. Dissolve 0.1 g of the substance under
heat until white fumes appear. Repeat the heating with a further examination, with warming, in 5 ml of chloroform and use the
5 ml of water, cool and add 40 ml of water and 10 ml of stannated warm solution.
hydrochloric acid AsT. The resulting solution complies with Reference solution. Dissolve 5 mg of (+)-menthol, 5 µl of
the limit test for arsenic (1 ppm). Prepare the standard using menthyl acetate and 5 mg of thymol in 10 ml of toluene.
0.5 ml of arsenic standard solution (10 ppm As).
Apply separately to the plate, as bands 20 mm x 3 mm, 30 µl of
Heavy metals (2.3.13). To the residue obtained in the test for test solution and 10 µl of reference solution.
Sulphated ash add 1 ml of hydrochloric acid, evaporate to
After development, dry the plate in air and spray with a freshly
dryness on a water-bath and dissolve the residue in 20 ml of
prepared 20 per cent w/v solution of phosphomolybdic acid
water. 12 ml of the solution complies with the limit test for
in ethanol (95 per cent) and heat at 105° for 15 minutes. The
heavy metals, Method D ( 20 ppm). Prepare the standard using
chromatogram obtained with the reference solution shows in
lead standard solution (1 ppm Pb).
the lower part a dark blue band due to menthol, a reddish band
Chlorides (2.3.12). 10 ml of solution A complies with the limit above it due to thymol and a dark blue band in the upper part
test for chlorides (0.25 per cent). due to menthyl acetate. The chromatogram obtained with the
test solution shows a large blue band due to triacontanol
Sulphated ash (2.3.18). 20.0 to 33.3 per cent, calculated on the
(melissyl alcohol) at an Rf value between those of the bands
dried basis, determined on 1.0 g dispersed in a mixture of
due to menthol and thymol in the chromatogram obtained
equal volumes of sulphuric acid and water.
with the reference solution and blue bands at Rf values
Loss on drying (2.4.19). Not more than 10.0 per cent, between those of the bands due to menthyl acetate and thymol
determined on 1.0 g by drying in an oven at 105°. in the chromatogram obtained with the reference solution. In
addition, the chromatogram obtained with the test solution
Assay. Weigh accurately about 0.2 g and disperse in 80 ml of
shows further bands at higher Rf values than menthyl acetate,
anhydrous glacial acetic acid. Heat on a water-bath for
that with the highest Rf value being very pronounced, and a
2 hours, cool. Titrate with 0.1 M perchloric acid determining
number of faint bands below that due to triacontanol; a band
the end-point potentiometrically (2.4.25). Carry out a blank
on the line of application is blue.
titration.
1 ml of 0.1 M perchloric acid is equivalent to 0.002299 g of Tests
Na. Melting range (2.4.21). 78° to 88°, determined by Method II.
Storage. Store protected from light. Heavy metals (2.3.13). 0.5 g complies with the limit test for
Labelling. The label states (1) the apparent viscosity in heavy metals, Method B (40 ppm).
millipascal seconds of a 2 per cent w/v solution or, where the Acid value. Not more than 12.0, determined by the following
viscosity is low, the concentration of the solution to be used method. Weigh accurately about 2.0 g (w) in a flask fitted with
and the apparent viscosity in mPa s; (2) that the contents are a reflux condenser, add 40 ml of xylene and heat until the
not intended for use in the manufacture of an injectable substance has dissolved. Add 20 ml of ethanol (95 per cent)
preparation. and titrate the hot solution with 0.5 M ethanolic potassium
244
IP 2007 CEFACLOR
hydroxide, using phenolphthalein solution as indicator, until Specific optical rotation (2.4.22). +101º to +111º, determined
a pink colour persists for at least 10 seconds (n1 ml). Repeat in 1.0 per cent w/v solution in a 1.0 per cent w/v solution of
the operation without the substance under examination hydrochloric acid.
(n 2 ml). Calculate the acid value from the expression Related substances. Determine by liquid chromatography
28.05(n2–n1)/w. (2.4.14).
Saponification value. Between 78 and 95, determined by the
Solvent mixture. A 0.27 per cent w/v solution of sodium
following method. To the titrated solution from the
dihydrogen phosphate, adjusted to pH 2.5 with phosphoric
determination of the Acid value, add 20.0 ml of 0.5 M ethanolic
acid.
potassium hydroxide and boil under a reflux condenser for
3 hours. Titrate the hot solution immediately with 0.5 M Test solution. Dissolve 50 mg of the substance under
hydrochloric acid, using 1 ml of phenolphthalein solution examination in 10 ml of the solvent mixture.
as indicator, until the red colour is discharged. Reheat the Reference solution (a) A solution containing 0.0025 per cent
solution to boiling and continue the titration, if necessary, w/v cefaclor RS and 0.005 per cent w/v delta-3-cefaclor RS in
until the red colour no longer reappears on heating (n3 ml). the solvent mixture.
Repeat the operation without the substance under examination
(n4 ml). Calculate the saponification value from the expression Reference solution (b) Dilute 1 ml of the test solution to 100
a+[28.05(n4-n3)/w] where a is the acid value. ml with the solvent mixture.
Sulphated ash (2.3.18). Not more than 0.25 per cent, determined Chromatographic system
on 2.0 g. – a stainless steel column 25 cm x 4.6 mm, packed with
Storage. Store protected from light and moisture. – mobile phase: A. a 0.78 per cent w/v solution of sodium
dihydrogen phosphate adjusted to pH 4.0 with
phosphoric acid,
B. mix 450 ml of acetonitrile with 550 ml
Cefaclor of mobile phase A,
COOH – a linear gradient programme using the conditions given
O Cl below,
NH2 H N – spectrophotometer set at 220 nm,
N , H 20 – a 20 µl loop injector.
S
H H Increase the concentration of mobile phase B continuously
O and linearly by 0.67 per cent v/v per minute for 30 minutes (25
per cent v/v). Then increase the concentration of mobile phase
C15H14ClN3O4S,H2O Mol. Wt 385.8 B continuously and linearly by 5 per cent v/v per minute for 15
minutes (100 per cent v/v). Finally elute with mobile phase B
Cefaclor is (6R,7R)-7-[[(2R)-2-amino-2-phenylacetyl]amino]-
for 10 minutes.
3-chloro-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
carboxylic acid. Equilibrate the column with a mixture of 5 volumes of mobile
phase B and 95 volumes of mobile phase A for at least 15
Cefaclor contains not less than 96.0 per cent and not more
minutes between each analysis. Inject the solutions. At the
than 102.0 per cent of C15H14ClN3O4S, calculated on the
end of the programme change the composition of the mobile
anhydrous basis.
phase to a mixture of 5 volumes of mobile phase B and 95
Description. A white or slightly yellow powder. volumes of mobile phase A to re-equilibrate the column.
Identification Inject reference solution (a). The test is not valid unless the
resolution between the peaks due to cefaclor and delta-3-
Determine by infrared absorption spectrophotometry (2.4.6). cefaclor is not less than 2.0 and the tailing factor of the cefaclor
Compare the spectrum with that obtained with cefaclor RS or peak is not more than 1.2. If necessary, adjust the acetonitrile
with the reference spectrum of cefaclor. content of the mobile phase.
Tests Inject the test solution and reference solution (b). In the
pH (2.4.24). 3.0 to 4.5, determined in a suspension, prepared peak, other than the principal peak and any peaks due to the
by dispersing 0.25 g in 10 ml of carbon dioxide-free water. mobile phase, is not greater than 0.5 times the area of the
245
CEFACLOR CAPSULES IP 2007
principal peak in the chromatogram obtained with reference Identification

solution (b) (0.5 per cent) and the sum of the areas of all such
peaks is not more than twice the area of the principal peak in A. Shake a quantity of the contents of the capsules containing
the chromatogram obtained with reference solution (b) (2.0 0.3 g of anhydrous cefaclor with 100 ml of water, filter and
per cent). Ignore any peak with an area less than 0.1 times the dilute 1 ml of the filtrate to 100 ml with water.
area of the principal peak in the chromatogram obtained with When examined in the range 190 nm to 310 nm (24.7), the
reference solution (b). resulting solution shows an absorption maximum only at about
Heavy metals (2.3.13). 1.0 g complies with the limit test for 264 nm.
heavy metals, Method B (20 ppm). B. In the Assay, the principal peak in the chromatogram
Water (2.3.43). 3.0 to 6.5 per cent, determined on 0.2 g. obtained with the test solution corresponds to the peak in the
Test solution. Dissolve 15 mg of the substance under Tests
examination in the mobile phase and dilute to 50.0 ml with the Dissolution (2.5.2).
mobile phase.
Apparatus. No 1
Reference solution (a). A 0.03 per cent w/v solution of cefaclor Medium. 900 ml of water.
RS in the mobile phase. Speed and time. 50 rpm and 45 minute.
Reference solution (b). A solution containing 0.03 per cent
w/v each of cefaclor RS and delta-3-cefaclor RS in the mobile
phase.
the medium if necessary, at the maximum at about 264 nm
Chromatographic system (2.4.7). Calculate the content of C15H14ClN3O4S in the medium
– a stainless steel column 25 cm x 4.6 mm, packed with from the absorbance obtained from a solution of known
octadecylsilane bonded to porous silica (5 µm), concentration of cefaclor RS in the same medium.
– mobile phase: a mixture prepared by adding 220 volumes
of methanol to a mixture of 780 volumes of water, 10
(2.4.14).
volumes of triethylamine and 1 g of sodium
pentanesulphonate, adjusted to pH 2.5 with Solvent mixture. A 0.27 per cent w/v solution of sodium
orthophosphoric acid, dihydrogen orthophosphate, adjusted pH to 2.5, if necessary,
– flow rate. 1.5 ml per minute, with orthophosphoric acid.
– spectrophotometer set at 265 nm, Test solution. Shake a quantity of the contents of the capsules
– a 20 µl loop injector. containing 0.5 g of anhydrous cefaclor with 200 ml of the
Inject reference solution (b). The test is not valid unless the solvent mixture, dilute to 250 ml with the solvent mixture and
resolution between the peaks due to cefaclor and delta-3- filter.
cefaclor is not less than 2.5. Adjust the concentration of Reference solution (a). A 0.002 per cent w/v solution of
methanol in the mobile phase, if necessary. The test is not cefaclor RS in the solvent mixture.
valid unless the tailing factor of the cefaclor peak is not more
than 1.5. Reference solution (b). A solution containing 0.0025 per cent
w/v of cefaclor RS and 0.005 per cent w/v of delta-3-cefaclor
Inject reference solution (a) 6 times. The test is not valid unless
the relative standard deviation of the peak area of cefaclor is
not more than 1.0 per cent. Chromatographic system
Inject alternately the test solution and reference solution (a). octadecylsilyl silica gel (5 µm) (such as Spherisorb
Calculate the content of C15H14ClN3O4S. ODS-2),
Storage. Store protected from moisture. – mobile phase: A. a 0.78 per cent w/v solution of sodium
dihydrogen orthophosphate adjusted to pH 4.0 with
orthophosphoric acid,
Cefaclor Capsules B. a mixture of 450 volumes of acetonitrile
and 550 volumes of mobile phase A.
Cefaclor Capsules contains not less than 95.0 per cent and – flow rate. 1 ml per minute,
not more than 105.0 per cent of the stated amount of cefaclor, – a linear gradient programme using the conditions given
C15H14ClN3O4S. below,
246
IP 2007 CEFACLOR ORAL SUSPENSION
– spectrophotometer set at 220 nm, – a 20 µl loop injector.

– a 20 µl loop injector. Inject reference solution (b). The test is not valid unless the
Time Mobile Mobile Comments resolution between the peaks due to cefaclor and delta-3-
phase A phase B cefaclor is not less than 2.5.
0 – 30 95 – 75 5 – 25 linear gradient
Calculate the content of C15H14ClN3O4S in the capsules.
30 – 45 75 – 0 25 – 100
45 – 55 0 100 isocratic
55 – 70 0 – 95 100 – 5 re-equilibration Labelling. The quantity of active ingredient is stated in terms
of the equivalent amount of anhydrous cefaclor.
Equilibrate the column with a mixture of 5 volumes of mobile
minutes.
Inject reference solution (b). The test is not valid unless the Cefaclor Oral Suspension
resolution between the peaks due to cefaclor and delta-3- Cefaclor Oral Suspension is a mixture consisting of Cefaclor
cefaclor is not less than 2.0. If necessary, adjust the proportion with buffering agents and other excipients. It contains a
of acetonitrile in the mobile phase. suitable flavouring agent. It is filled in a sealed container.
Inject the test solution and reference solution (a). In the The suspension is constituted by dispersing the contents of
chromatogram obtained with the test solution the area of any the sealed container in the specified volume of Water just
secondary peak is not greater than half the area of the principal before use.
(a) (0.5 per cent) and the sum of the areas of any such peaks is Cefaclor Oral Suspension contains not less than 80.0 per cent
not greater than twice the area of the principal peak in the and not more than 120.0 per cent of the stated amount of
chromatogram obtained with reference solution (b) (2 per cefaclor, C15H14ClN3O4S.
cent). Ignore any peak with an area less than 0.1 times the area When stored at the temperature and for the period stated on
of the principal peak in the chromatogram obtained with the label during which the constituted suspension may be
reference solution (a) (0.1 per cent). expected to be satisfactory for use, it contains not less than
Other tests. Comply with the tests stated under Tablets. 80.0 per cent of the stated amount of cefaclor, C15H14ClN3O4S.
Assay. Determine by liquid chromatography (2.4.14). Storage. Store protected from moisture at a temperature not
exceeding 30º.
Test solution. Shake a quantity of the contents of capsules
containing about 75 mg of anhydrous cefaclor with 200 ml of The constituted suspension complies with the tests stated
the mobile phase, dilute to 250.0 ml with the mobile phase and under Oral liquids and with the following tests.
filter.
Identification
Reference solution (a). A 0.03 per cent w/v solution of cefaclor
RS in the mobile phase. A. Shake a quantity of the oral suspension containing 0.3 g of
Reference solution (b). A solution containing 0.03 per cent w/ anhydrous cefaclor with 500 ml of water and filter.
v each of cefaclor RS and delta-3-cefaclor RS in the mobile When examined in the range 190 nm to 310 nm (2.4.7), the
phase. filtrate shows an absorption maximum only at about 264 nm.
Chromatographic system B. In the Assay, the principal peak in the chromatogram
– a stainless steel column 25 cm x 4.6 mm, packed with obtained with the test solution corresponds to the peak in the
octadecylsilyl silica gel (5 µm) (such as Beckman chromatogram obtained with reference solution (a).
Ultrasphere ODS and Supelcosil LC-18-DB),
– mobile phase: a solution prepared by dissolving 1 g of Tests
sodium pentanesulphonate in a mixture of 780 volumes
of water and 10 volumes of triethylamine, adjusting
(2.4.14).
the pH to 2.5 with orthophosphoric acid adding 220
volumes of methanol and mixing, Solvent mixture. A 0.27 per cent w/v solution of sodium
– flow rate. 1.5 ml per minute, dihydrogen orthophosphate, adjusted to pH 2.5 with
– spectrophotometer set at 265 nm, orthophosphoric acid.
247
CEFACLOR SUSTAINED-RELEASE TABLETS IP 2007
Test solution. Shake a quantity of the oral suspension Assay. Determine by liquid chromatography (2.4.14).
containing about 0.25 g of anhydrous cefaclor with 200 ml of Test solution. Shake a quantity of the oral suspension
the solvent mixture, dilute to 250 ml with the solvent mixture containing about 75 mg of anhydrous cefaclor with 200 ml of
and filter. the mobile phase, dilute to 250.0 ml with the mobile phase and
Reference solution (a). A 0.001 per cent w/v solution of filter.
cefaclor RS in the solvent mixture. Reference solution (a). A 0.03 per cent w/v solution of cefaclor
Reference solution (b). A solution containing 0.0025 per cent RS in the mobile phase.
w/v of cefaclor RS and 0.005 per cent w/v of delta-3-cefaclor Reference solution (b). A solution containing 0.03 per cent w/
RS in the solvent mixture. v each of cefaclor RS and delta-3-cefaclor RS in the mobile
Chromatographic system phase.
– a stainless steel column 25 cm x 4.6 mm, packed with Chromatographic system
octadecylsilyl silica gel (5 µm) (such as Spherisorb – a stainless steel column 25 cm x 4.6 mm, packed with
ODS-2), octadecylsilyl silica gel (5 µm) (such as Beckman
– mobile phase: A. a 0.78 per cent w/v solution of sodium Ultrasphere ODS and Supelcosil LC-18-DB),
dihydrogen orthophosphate,djusted to pH 4.0 with – mobile phase: a solution prepared by dissolving 1 g of
orthophosphoric acid, sodium pentanesulphonate in a mixture of 780 ml of
B. a mixture of 450 volumes of acetonitrile water and 10 ml of triethylamine, adjusting the pH to
and 550 volumes of mobile phase A, 2.5 using orthophosphoric acid, adding 220 ml of
– flow rate. 1 ml per minute, methanol and mixing,
– a linear gradient programme using the conditions given – flow rate. 1.5 ml per minute,
below, – spectrophotometer set at 265 nm,
Time Mobile Mobile Comments resolution between the peaks due to cefaclor and delta-3-
phase A phase B cefaclor is not less than 2.5 and the tailing factor of the peak
(in min.) (per cent v/v) (per cent v/v) due to cefaclor is not more than 1.5.
0 – 30 95 – 75 5 – 25 linear gradient Inject alternately the test solution and reference solution (a).
30 – 45 75 – 0 25 – 100 linear gradient
Determine the weight per ml of the oral suspension (2.4.29)
45 – 55 0 100 isocratic and calculate the content of C15H14ClN3O4S, weight in volume.
55 – 70 0 – 95 100 – 5 re-equilibration Repeat the procedure using a portion of the constituted
Equilibrate the column with a mixture of 5 volumes of mobile suspension that has been stored at the temperature and for
phase B and 95 volumes of mobile phase A for at least 15 the period stated on the label.
minutes. Storage. Store at the temperature and use within the period
Inject reference solution (b). The test is not valid unless stated on the label.
resolution between the peaks due to cefaclor and delta-3- Labelling. The label states the quantity in terms of the
cefaclor is not less than 2.0. If necessary, adjust the proportion equivalent amount of anhydrous cefaclor.
of acetonitrile in the mobile phase.
Inject the test solution and reference solution (a). In the
secondary peak is not greater than the area of the principal
Cefaclor Sustained-release Tablets
(a) (1 per cent) and the sum of the areas of any such peaks is Cefaclor Sustained-release Tablets are prolonged-release
not greater than three times the area of the principal peak in tablets containing Cefaclor. The appropriate release of the
the chromatogram obtained with reference solution (a) (3 per active ingredient is demonstrated by a suitable dissolution
cent). Ignore any peak with an area less than 0.1 times the area test
of the principal peak in the chromatogram obtained with
Cefaclor Tablets contains not less than 90.0 per cent and not
reference solution (a) (0.1 per cent).
more than 105.0 per cent of the stated amount of cefaclor,
Other tests. Complies with the tests stated under Oral Liquids. C15H14ClN3O4S.
248
IP 2007 CEFACLOR SUSTAINED-RELEASE TABLETS
Identification Equilibrate the column with a mixture of 5 volumes of mobile

A. Shake a quantity of the powdered tablets containing 0.3 g minutes.
of anhydrous cefaclor with 100 ml of water, filter and dilute 1
ml of the filtrate to 100 ml with water. Inject reference solution (b). The test is not valid unless the
When examined in the range 190 nm to 310 nm (2.4.7), the cefaclor is not less than 2.0. If necessary, adjust the proportion
resulting solution shows an absorption maximum only at about of acetonitrile in the mobile phase.
264 nm.
Inject the test solution and reference solution (a). In the
chromatogram obtained with test solution the area of any
secondary peak is not greater than 0.6 times the area of the
solution (a) (0.6 per cent) and the sum of the areas of any such
Tests
peaks is not greater than twice the area of the principal peak in
Related substances. Determine by liquid chromatography the chromatogram obtained with reference solution (a) (2 per
(2.4.14). cent). Ignore any peak with an area less than 0.1 times the area
of the principal peak in the chromatogram obtained with
Solvent mixture. A 0.27 per cent w/v solution of sodium
dihydrogen orthophosphate, adjusted pH to 2.5, if necessary,
with orthophosphoric acid. Assay. Determine by liquid chromatography (2.4.14).
Test solution. Shake a quantity of the powdered tablets Test solution. Weigh and powder 20 tablets. Weigh accurately
containing 0.75 g of anhydrous cefaclor with 200 ml of the a quantity of the powder containing about 75 mg of anhydrous
solvent mixture, dilute to 250 ml with the solvent mixture and cefaclor, disperse in the mobile phase, shake, dilute to 250.0 ml
filter. with the mobile phase and filter.
Reference solution (a). A 0.003 per cent w/v solution of Reference solution (a). A 0.03 per cent w/v solution of cefaclor
cefaclor RS in the solvent mixture. RS in the mobile phase.
Reference solution (b). A solution containing 0.0025 per cent Reference solution (b). A solution containing 0.03 per cent w/
w/v of cefaclor RS and 0.005 per cent w/v of delta-3-cefaclor v each of cefaclor RS and delta-3-cefaclor RS in the mobile
RS in the solvent mixture. phase.
Chromatographic system Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilyl silica gel (5 µm) (such as Spherisorb ODS- octadecylsilyl silica gel (5 µm) (such as Beckman
2), Ultrasphere ODS and Supelcosil LC-18-DB),
– mobile phase: A. a 0.78 per cent w/v solution of sodium – mobile phase: a solution prepared by dissolving 1 g of
dihydrogen orthophosphate adjusted to pH 4.0 with sodium pentanesulphonate in a mixture of 780 volumes
orthophosphoric acid of water and 10 volumes of triethylamine, adjusting
B. a mixture of 450 volumes of acetonitrile the pH to 2.5 with orthophosphoric acid, adding 220
and 550 volumes of mobile phase A. volumes of methanol and mixing,
– flow rate. 1 ml per minute, – flow rate. 1.5 ml per minute,
– a linear gradient programme using the conditions given – spectrophotometer set at 265 nm,
below, – a 20 µl loop injector.
Time Mobile Mobile Comment cefaclor is not less than 2.5.
phase A phase B
(in min.) (per cent v/v) (per cent v/v) Inject the test solution and reference solution (a).
0 – 30 95 – 75 5 – 25 linear gradient Calculate the content of C15H14ClN3O4S in the tablets.
30 – 45 75 – 0 25 – 100 linear gradient Storage. Store protected from moisture.
45 – 55 0 100 isocratic Labelling. The label states the strength in terms of the
55 – 70 0 – 95 100 – 5 re-equilibration equivalent amount of anhydrous cefaclor.
249
CEFADROXIL IP 2007
Cefadroxil Specific optical rotation (2.4.22). +165° to +178°, determined

COOH Related substances. Determine by thin-layer chromatography
HO O CH3 (2.4.17), coating the plate with silica gel G.
O N ,H2O
Mobile phase. A mixture of 56 volumes of ethyl acetate, 20
N S volumes of ethanol (95 per cent), 20 volumes of water and
NH2 H H H 4 volumes of formic acid.
C16H17N3O5S,H2O Mol. Wt. 381.4 Solvent mixture. 75 volumes of ethanol (95 per cent),
22 volumes of water and 3 volumes 2.4 M hydrochloric acid.
Cefadroxil is 7-[(R)-2-amino-2-(4-hydroxyphenyl)acetamido]-
3-methyl-3-cephem-4-carboxylic acid monohydrate. Test solution. A 2.5 per cent w/v solution of the substance
under examination in the solvent mixture.
Cefadroxil contains not less than 95.0 per cent and not more
than 101.0 per cent of C16H 17N 3O 5S, calculated on the Reference solution (a). Dilute 1 ml of the test solution to
anhydrous basis. 100 ml with the solvent mixture.
Description. A white to off-white, crystalline powder. Reference solution (b). A solution containing 0.025 per cent
w/v each of 7-aminodesacetoxycephasporanic acid RS and
Identification D-a-4-hydroxyphenylglycine RS.
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution (c). A mixture of equal volumes of the test
Compare the spectrum with that obtained with cefadroxil RS solution and reference solution (b).
or with the reference spectrum of cefadroxil. Apply to the plate 2 µl of the test solution, reference solution
B. Determine by thin-layer chromatography (2.4.17), coating (a) and reference solution (b) and 4 µl of solution (c). Allow
the plate with silica gel H and impregnating the dry plate by the mobile phase to rise 12 cm. Dry the plate in air, and spray
placing it in a tank containing a shallow layer of about 1 cm of with a 3.0 per cent w/v solution of ninhydrin in a 4.55 per cent
a mixture of 95 volumes of n-hexane and 5 volumes of w/v solution of sodium metabisulphite. Any secondary spot
1-tetradecane, allowing the solvent to ascend to the top, in the chromatogram obtained with the test solution is not
removing the plate and allowing the solvent to evaporate. more intense than any of the spots in the chromatograms
obtained with reference solution (a) and reference solution
Mobile phase. A mixture of 60 volumes of 0.1 M citric acid, (b). The test is not valid unless the chromatogram obtained
40 volumes of 0.1 M disodium hydrogen phosphate and with reference solution (c) exhibits three distinct spots.
1.5 volumes of a 6.66 per cent w/v solution of ninhydrin in
acetone. N,N-Dimethylaniline (2.3.21). Not more than 20 ppm,
determined by Method B.
under examination in water. Water (2.3.43). 4.2 to 6.0 per cent, determined on 0.5 g.
Reference solution (a). A 0.2 per cent w/v solution of cefadroxil Assay. Determine by liquid chromatography (2.4.14).
RS in water. Test solution. A freshly prepared 0.1 per cent w/v solution of
Reference solution (b). A mixture of equal volumes of the test the substance under examination in phosphate buffer pH 5.0.
solution and reference solution (a). Reference solution. A freshly prepared 0.1 per cent w/v solution
Apply to the plate 20 µl of each solution After development, of cefadroxil RS in phosphate buffer pH 5.0.
dry the plate in air, spray with a 0.2 per cent w/v solution of Chromatographic system
ninhydrin in ethanol, dry at 110° for 10 minutes and examine. – a stainless steel column 25 cm x 4 mm, packed with
The principal spot in the chromatogram obtained with the test octadecylsilyl silica gel (3 to 10 µm),
solution corresponds to that in the chromatogram obtained – mobile phase: a mixture of 96 volumes of phosphate
with reference solution (a). The principal spot in the buffer pH 5.0 and 4 volumes of acetonitrile,
chromatogram obtained with reference solution (b) appears – flow rate. 1.5 ml per minute,
as a single compact spot. – spectrophotometer set at 230 nm,
Tests
pH (2.4.24). 4.0 to 6.0, determined in a 5.0 per cent w/v relative standard deviation for replicate injections is not more
suspension. than 2.0 per cent.
250
IP 2007 CEFADROXIL ORAL SUSPENSION
Inject alternately the test solution and the reference solution. Calculate the content of C16H17N3O5S in the medium from the
Calculate the content of C16H17N3O5S. absorbance obtained from a solution of known concentration
of cefadroxil RS.
exceeding 30°.
C16H17N3O5S.
Cefadroxil Capsules Assay. Determine by liquid chromatography (2.4.14).
Cefadroxil Capsules contain not less than 90.0 per cent and Prepare the following solutions freshly.
not more than 120.0 per cent of the stated amount of Test solution. Weigh accurately a quantity of the mixed
anhydrous cefadroxil, C16H17N3O5S. contents of 20 capsules containing about 0.2 g of Cefadroxil,
add sufficient phosphate buffer pH 5.0, shake for 30 minutes,
Identification
dilute to 200.0 ml with the same solvent and filter.
Determine by thin-layer chromatography (2.4.17), coating the Reference solution. A 0.1 per cent w/v solution of cefadroxil
plate with silica gel H and impregnating the dry plate by RS in phosphate buffer pH 5.0.
placing it in a tank containing a shallow layer of about 1 cm of
a mixture of 95 volumes of n-hexane and 5 volumes of Chromatographic system
1-tetradecane, allowing the solvent to ascend to the top, – a stainless steel column 25 cm x 4 mm, packed with
removing the plate and allowing the solvent to evaporate. octadecylsilyl silica gel (5 µm),
– mobile phase: a mixture of 96 volumes of phosphate
Mobile phase. A mixture of 60 volumes of 0.1 M citric acid, 40 buffer pH 5.0 and 4 volumes of acetonitrile,
volumes of 0.1 M disodium hydrogen phosphate and – flow rate. 1.5 ml per minute,
1.5 volumes of a 6.66 per cent w/v solution of ninhydrin in – spectrophotometer set at 230 nm,
acetone. – a 20 µl loop injector.
Test solution. Shake a quantity of the contents of a capsule
with sufficient water to produce a solution containing 0.2 per
cent w/v of Cefadroxil.
than 2.0 per cent.
cefadroxil RS in water.
Calculate the content of C16H17N3O5S in the capsules.
solution and reference solution (a). Storage. Store protected from moisture at a temperature not
exceeding 30°.
dry the plate in air, spray with a 0.2 per cent w/v solution of Labelling. The label states the strength in terms of anhydrous
ninhydrin in ethanol, dry at 110° for 10 minutes and examine. cefadroxil.
solution corresponds to that in the chromatogram obtained
with reference solution (a). The principal spot in the Cefadroxil Oral Suspension
chromatogram obtained with reference solution (b) appears
as a single compact spot. Cefadroxil Mixture
Cefadroxil Oral Suspension is a mixture of Cefadroxil with
Tests
buffering agents and other excipients. It contains a suitable
Water (2.3.43). Not more than 7.0 per cent, determined on flavouring agent. It is filled in a sealed container.
0.5 g of the mixed contents of 20 capsules.
Dissolution (2.5.2). the sealed container in the specified volume of water just
Apparatus. No 1 before use.
Medium. 900 ml of water. Cefadroxil Oral Suspension contains not less than 90.0 per
Speed and time. 100 rpm and 45 minutes. cent and not more than 120.0 per cent of the stated amount of
Withdraw a suitable volume of the medium and filter. Measure C16H17N3O5S.
the absorbance (2.4.7) of the filtrate, suitably diluted if When stored at the temperature and for the period stated on
necessary, at the maximum at about 263 nm. the label during which the constituted suspension may be
251
CEFADROXIL TABLETS IP 2007
expected to be satisfactory for use, it contains not less than – mobile phase: a mixture of 96 volumes of phosphate
80.0 per cent of the stated amount of cefadroxil. buffer pH 5.0 and 4 volumes of acetonitrile,
Determine by thin-layer chromatography (2.4.17), coating the
plate with silica gel H and impregnating the dry plate by Inject the reference solution. The test is not valid unless the
placing it in a tank containing a shallow layer of about 1 cm of relative standard deviation for replicate injections is not more
a mixture of 95 volumes of n-hexane and 5 volumes of than 2.0 per cent.
1-tetradecane, allowing the solvent to ascend to the top, Inject alternately the test solution and the reference solution.
removing the plate and allowing the solvent to evaporate.
Determine the weight per ml (2.4.29) of the suspension and
Mobile phase. A mixture of 60 volumes of 0.1 M citric acid,
calculate the content of C16H17N3O5S, weight in volume.
40 volumes of 0.1 M disodium hydrogen phosphate and
1.5 volumes of a 6.66 per cent w/v solution of ninhydrin in Repeat the procedure using a portion of the suspension that
acetone. has been stored at the temperature and for the period stated
on the label during which it may be expected to be satisfactory
Test solution. Dilute a suitable quantity of the freshly prepared
for use.
suspension with water to obtain a solution containing 0.2 per
cent w/v of cefadroxil. Filter the solution. Storage. Store protected from moisture, at a temperature not
exceeding 30°.
Reference solution (a). A 0.2 per cent w/v solution of cefadroxil
RS in water. Labelling. The label states the quantity of active ingredient in
terms of anhydrous cefadroxil.
Apply to the plate 20 µl of each solution After development,
dry the plate in air, spray with a 0.2 per cent w/v solution of Cefadroxil Tablets
ninhydrin in ethanol, dry at 110° for 10 minutes and examine. Cefadroxil Tablets contain not less than 90.0 per cent and
The principal spot in the chromatogram obtained with the test not more than 120.0 per cent of the stated amount of
solution corresponds to that in the chromatogram obtained anhydrous cefadroxil, C16H17N3O5S. The tablets may be
with reference solution (a). The principal spot in the coated.
chromatogram obtained with reference solution (b) appears
as a single compact spot. Identification
Water (2.3.43). Not more than 2.0 per cent, determined on Determine by thin-layer chromatography (2.4.17), coating the
1.0 g, using a mixture of 2 volumes of carbon tetrachloride, plate with silica gel H and impregnating the dry plate by
2 volumes of chloroform and 1 volume of methanol in place of placing it in a tank containing a shallow layer of about 1 cm of
methanol in the titration vessel. a mixture of 95 volumes of n-hexane and 5 volumes of
The constituted suspension complies with the tests stated 1-tetradecane, allowing the solvent to ascend to the top,
under Oral liquids and with the following tests. removing the plate and allowing the solvent to evaporate.
Mobile phase. A mixture of 60 volumes of 0.1 M citric acid,
Tests
40 volumes of 0.1 M disodium hydrogen phosphate and
pH (2.4.24). 4.5 to 6.0. 1.5 volumes of a 6.66 per cent w/v solution of ninhydrin in
acetone.
Test solution. Shake a quantity of the powdered tablets with
Test solution. Transfer an accurately weighed quantity of the
sufficient water to produce a solution containing 0.2 per cent
suspension containing about 0.1g of cefadroxil to a 100-ml
w/v of cefadroxil. Filter the solution.
volumetric flask, add phosphate buffer pH 5.0, shake for
30 minutes, dilute to 100.0 ml with the same solvent and filter. Reference solution (a). A 0.2 per cent w/v solution of cefadroxil
RS in water.
Reference solution. A 0.1 per cent w/v solution of cefadroxil
RS in phosphate buffer pH 5.0. Reference solution (b). A mixture of equal volumes of the test
Chromatographic system solution and reference solution (a).
– a stainless steel column 25 cm x 4 mm, packed with Apply to the plate 20 µl of each solution After development,
octadecylsilyl silica gel (5 µm), dry the plate in air, spray with a 0.2 per cent w/v solution of
252
IP 2007 CEFAZOLIN SODIUM
ninhydrin in ethanol, dry at 110° for 10 minutes and examine. Cefazolin Sodium
solution corresponds to that in the chromatogram obtained Cephazolin Sodium
with reference solution (a). The principal spot in the
chromatogram obtained with reference solution (b) appears COONa N N
as a single compact spot. O CH3
N N O N S S
Tests N N
N S
Dissolution (2.5.2). H H H
Apparatus. No 1
Medium. 900 ml of water. C14H13N8NaO4S3 Mol. Wt. 476.5
Speed and time. 50 rpm and 30 minutes. Cefazolin Sodium is sodium 7-[(1H)-tetrazol-1-ylacetamido]-
Withdraw a suitable volume of the medium and filter. Measure 3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-
the absorbance of the filtrate, suitably diluted if necessary, at carboxylate.
the maximum at about 263 nm (2.4.7). Calculate the content of Cefazolin Sodium contains not less than 85.0 per cent and not
C16H17N3O5S in the medium from the absorbance obtained from more than 105.0 per cent of cefazolin C14H14N8O4S3, calculated
a solution of known concentration of cefadroxil RS. on the anhydrous basis.
D. Not less than 75 per cent of the stated amount of Description. A white to off-white, crystalline powder;
C16H17N3O5S. odourless.
Water (2.3.43). Not more than 8.0 per cent, determined on Identification
0.5 g of the powdered tablets. A. Determine by infrared absorption spectrophotometry (2.4.6).
Assay. Determine by liquid chromatography (2.4.14). Compare the spectrum with that obtained with cefazoline
sodium RS or with the reference spectrum of cefazolin sodium.
Prepare the following solutions freshly.
a quantity of the powder containing about 0.2 g of cefadroxil,
dissolve in phosphate buffer pH 5.0 by shaking for 30 minutes
and dilute to 200.0 ml the same solvent. Filter the solution. Tests
Reference solution. A 0.1 per cent w/v solution of cefadroxil
RS in phosphate buffer pH 5.0. pH (2.4.24). 4.0 to 6.0, determined in a 10.0 per cent w/v solution.
Chromatographic system Specific optical rotation (2.4.22). -10.0° to -24.0°, determined

– a stainless steel column 25 cm x 4 mm, packed with in a 5.5 per cent w/v solution in 0.1 M sodium bicarbonate.
octadecylsilyl silica gel (3 to 10 µm), Water (2.3.43). Not more than 6.0 per cent, determined on
– mobile phase: a mixture of 96 volumes of phosphate 0.15 g.
buffer pH 5.0 and 4 volumes of acetonitrile,
– spectrophotometer set at 230 nm, Solution A. Prepared by dissolving 0.75 g of salicylic acid
– a 20 µl loop injector. (internal standard) in 5 ml of methanol and diluting to 100.0 ml
with mixed phosphate buffer pH 7.0.
relative standard deviation for replicate injections is not more Test solution. A 0.1 per cent w/v solution of the substance
than 2.0 per cent. under examination in mixed phosphate buffer pH 7.0. to 5.0 ml
of this solution add 5.0 ml of solution A and add sufficient
volume of mixed phosphate buffer pH 7.0 to produce 100.0 ml
Calculate the content of C16H17N3O5S in the tablets. and mix.
Storage. Store protected from moisture at a temperature not Reference solution. A 0.1 per cent w/v solution of cefazolin
exceeding 30°. sodium RS in mixed phosphate buffer pH 7.0. To 5.0 ml of this
Labelling. The label states the strength in terms of anhydrous solution add 5.0 ml of solution A and add sufficient volume of
cefadroxil. mixed phosphate buffer pH 7.0 to produce 100.0 ml and mix.
253
CEFAZOLIN SODIUM INJECTION IP 2007
Chromatographic system Description. A white to off-white, crystalline powder;

– a stainless steel column 30 cm x 4 mm, packed with odourless.
octadecylsilyl silica gel (3 to 10 µm), The contents of the sealed container comply with the
– mobile phase: a mixture of 9 volumes of phosphate buffer requirements stated under Parenteral Preparations
pH 3.6 and 1 volume of acetonitrile, (Powders for Injection) and with the following requirements.
– spectrophotometer set at 254 nm, Identification
A. Determine by infra-red absorption spectrophotometry
Inject the reference solution. The test is not valid unless the (2.4.6). Compare the spectrum with that obtained with
relative retention times of salicylic acid and cefazolin are 0.7 cefazoline sodium RS or with the reference spectrum of
and 1.0 respectively. cefazolin sodium.
Inject alternately the test solution and the reference solution. B. In the Assay, the principal peak in the chromatogram
Calculate the content of C14H14N8O4S3. obtained with the test solution corresponds to the peak in the
Cefazolin Sodium intended for use in the manufacture of
parenteral preparations complies with the following Tests
additional requirements.
Specific optical rotation (2.4.22). -10.0° to -24.0°, determined
Unit per mg of cefazolin.
in a 5.5 per cent w/v solution in 0.1 M sodium bicarbonate.
Sterility (2.2.11). Complies with the test for sterility. Bacterial endotoxins (2.2.3). Not more than 0.15 Endotoxin
Storage. Store in sterile containers, sealed so as to exclude Unit per mg of cefazolin.
micro-organisms protected from moisture at a temperature not Water (2.3.43). Not more than 6.0 per cent, determined on 0.15 g.
exceeding 30°.
Labelling. The label states the quantity of Cefazolin Sodium
contained in the sealed container in terms of the equivalent Solution A. Prepare by dissolving 0.75 g of salicylic acid
amount of cefazolin. (internal standard) in 5 ml of methanol and diluting to 100.0 ml
with mixed phosphate buffer pH 7.0.
Test solution. Determine the weight of the contents of 10
containers. Weigh accurately a quantity of the mixed contents
Cefazolin Sodium Injection of the 10 containers, dissolve in the mixed phosphate buffer
pH 7.0 and dilute to obtain a solution containing 0.1 per cent
Cefazolin Injection; Cephazolin Sodium Injection; w/v of cefazolin. To 5.0 ml of this solution add 5.0 ml of solution
Cephazolin Injection A and add sufficient volume of mixed phosphate buffer pH
Cefazolin Sodium Injection is a sterile material consisting of 7.0 to produce 100.0 ml and mix.
Cefazolin Sodium with or without excipients. It is filled in sealed Reference solution. A 0.1 per cent w/v solution of cefazolin
containers. sodium RS in mixed phosphate buffer pH 7.0. To 5.0 ml of this
The injection is constituted by dissolving the contents of the solution add 5.0 ml of solution A and sufficient volume of
sealed container in the requisite amount of sterile Water for mixed phosphate buffer pH 7.0 to produce 100.0 ml and mix.
Injections, immediately before use. Chromatographic system
The constituted solution complies with the requirements for – a stainless steel column 30 cm x 4 mm, packed with
Clarity of solution and Particulate matter stated under octadecylsilyl silica gel (3 to 10 µm),
Parenteral Preparations (Injections). – mobile phase: a mixture of 9 volumes of phosphate buffer
pH 3.6 and 1 volume of acetonitrile,
Storage. The constituted solution should be used immediately – flow rate. 2 ml per minute,
after preparation but, in any case, within the period – spectrophotometer set at 254 nm,
recommended by the manufacturer. – a 20 µl loop injector.
Cefazolin Sodium Injection contains not less than 90.0 per Inject the reference solution. The test is not valid unless the
cent and not more than 115.0 per cent of the stated amount of relative retention times of salicylic acid and cefazolin are 0.7
cefazolin, C14H14N8O4S. and 1.0 respectively.
254
IP 2007 CEFOPERAZONE INJECTION
Inject alternately the test solution and the reference solution. Reference solution. A 0.01 per cent w/v solution of
Calculate the content of C14H14N8O4S3 in the injection. cefoperazone dihydrate RS in the mobile phase.
exceeding 30°. The constituted solution should be stored
protected from light and used within 24 hours when stored at
a temperature not exceeding 30° or within 4 days when stored
volumes of acetonitrile, 3.5 volumes of a 6 per cent w/
between 2° and 8°.
v solution of acetic acid and 2.5 volumes of a solution
Labelling. The label states the quantity of Cefazolin Sodium prepared by dissolving 14 ml of triethylamine and 5.7
contained in the sealed container in terms of the equivalent ml of glacial acetic acid in 100 ml of water, and mixed,
amount of cefazolin. – flow rate. 2 ml per minute,
Cefoperazone Sodium Inject the reference solution. The test is not valid unless the
theoretical plates is not less than 5000, the symmetry factor is
CH3 at most 1.6 and the relative standard deviation for replicate
COONa
N injections is not more than 2.0 per cent.
H3C N H O O N
N S Inject alternately the test solution and the reference solution.
N N N N
O N Calculate the content of C25H26N9NaO8S2 by multiplying the
O O H H H S content of cefoperazone by 1.034.
Cefoperazone Sodium intended for use in the manufacture of
OH procedure for the removal of bacterial endotoxins complies
C25H26N9NaO8S2 Mol Wt. 667.7
Cefoperazone sodium is sodium salt of 7-D-(-)-α-(4-ethyl- Unit per mg of cefoperazone sodium.
2,3-dioxo-1-piperazinecarboxamido)-α-(4-hydroxyphenyl)
Cefoperazone Sodium intended for use in the manufacture of
acetamido-3-[(1-methyl-1H-tetrazol-5-yl)thio]methyl-3-
cephem-4-carboxylic acid.
sterilisation procedure complies with the following
Cefoperazone Sodium contains not less than 95.0 per cent additional requirement.
and not more than 102.0 per cent of C25H26N9NaO8S2, calculated
Description. A white or almost white crystalline powder.
Labelling. The label states whether it is intended for use in
Identification the manufacture of parenteral preparations.
A. In the Assay, the principal peak in the chromatogram
chromatogram obtained with the reference solution. Cefoperazone Injection
B. Gives the reactions of sodium salts (2.3.1). Cefoperazone Sodium Injection
Tests Cefoperazone Injection is a sterile material consisting of
Cefoperazone Sodium with or without excipients. It is filled in
a sealed container.
0.5 g.
Assay. Determine by liquid chromatography (2.4.14). Injections, immediately before use.
Test solution. Dissolve 10 mg of the substance under The constituted solution complies with the requirements for
examination in the mobile phase and dilute to 100.0 ml with the Clarity of solution and Particulate matter stated under
mobile phase. Parenteral Preparations (Injections).
255
CEFOTAXIME SODIUM IP 2007
Storage. The constituted solution should be used immediately Inject alternately the test solution and the reference solution.
after preparation but, in any case, within the period Calculate the content of C25H27N9O8S2 in the injection.
Cefoperazone Injection contains not less than 90.0 per cent exceeding 30º.
cefoperazone, C25H27N9O8S2. Labelling. The label states the quantity of Cefoperazone
Sodium contained in the sealed container in terms of the
Description. A white or almost white powder. equivalent amount of cefoperazone.
Cefotaxime Sodium
Identification
A. In the Assay, the principal peak in the chromatogram COONa O
H 2N
obtained with the test solution corresponds to the peak in the O
chromatogram obtained with the reference solution. N O N O CH3
S
B. It gives the reactions of sodium salts (2.3.1). N S
N H H H
Tests H3CO
pH (2.4.24). 4.5 to 6.5, determined in a 25.0 per cent w/v solution. C16H16N5NaO7S2 Mol. Wt. 477.4
Bacterial endotoxins (2.2.3). Not more than 0.20 Endotoxin Cefotaxime Sodium is sodium (7R)-3-acetoxymethyl-7-[(Z)-2-
Unit per mg of cefoperczone. (2-aminothiazol-yl)-2-(methoxyimino)acetamido]-3-cepham-
Water (2.3.43). Not more than 5.0 per cent, except that where it 4-carboxylate.
is in the freeze-dried form, the limit is not more than 2.0 per Cefotaxime Sodium contains the equivalent of not less than
cent. 85.5 per cent and not more than 96.4 per cent of cefotaxime,
Assay. Determine by liquid chromatography (2.4.14). C16H17N5O7S2, calculated on the anhydrous basis.
Test solution. Determine the weight of the contents of 10 Description. An off-white to pale yellow, crystalline powder.
of 10 containers containing about 25 mg of cefoperazone, Identification
dissolve in the mobile phase and dilute to 250.0 ml with the A. In the Assay, the principal peak in the chromatogram
mobile phase. obtained with the test solution corresponds to the peak in the
Reference solution. A 0.01 per cent w/v solution of chromatogram obtained with the reference solution.
cefoperazone dihydrate RS in the mobile phase. B. Gives the reactions of sodium salts (2.3.1).
– a stainless steel column 30 cm x 4.0 mm, packed with Tests
octadecylsilane bonded to porous silica (5 µm), pH (2.4.24). 4.5 to 6.5, determined in a 10.0 per cent w/v solution.
volumes of acetonitrile, 3.5 volumes of a 6 per cent w/ Water (2.3.43). Not more than 6.0 per cent, determined on
v solution of acetic acid and 2.5 volumes of a solution 0.15 g.
prepared by dissolving 14 ml of triethylamine and 5.7 Assay. Determine by liquid chromatography (2.4.14).
ml of glacial acetic acid in 100 ml of water, and mixed,
– flow rate. 2 ml per minute, Test solution. A 0.01 per cent w/v solution of the substance
– spectrophotometer set at 254 nm, under examination in water.
– a 10 µl loop injector. Reference solution. A 0.01 per cent w/v solution of cefotaxime
Inject the reference solution. The test is not valid unless the sodium RS in water.
theoretical plates is not less than 5000, the symmetry factor is Chromatographic system
at most 1.6 and the relative standard deviation for replicate – a stainless steel column 30 cm x 3.9 mm, packed with
injections is not more than 2.0 per cent. octadecylsilyl silica gel (3 to 10 µm),
256
IP 2007 CEFOTAXIME SODIUM INJECTION
– mobile phase: a solution prepared by dissolving 60 mg than 115.0 per cent of the stated amount of cefotaxime,
of potassium dihydrogen phosphate and 1.2 g of C16H17N5O7S2.
disodium hydrogen phosphate in 1000 ml of water and
Description. An off-white to pale yellow, crystalline powder.
mixing with 120 ml of methanol,
– flow rate. 1.5 ml per minute, The contents of the sealed container comply with the
– spectrophotometer set at 254 nm, requirements stated under Parenteral Preparations
– a 20 µl loop injector. (Powders for Injection) and with the following requirements.
relative standard deviation for replicate injections is not more Identification
than 2.0 per cent.
Inject alternately the test solution and the reference solution. obtained with the test solution corresponds to the peak in the
Calculate the content of C16H17N5O7S2.
B. Gives the reactions of sodium salts (2.3.1).
Cefotaxime Sodium intended for use in the manufacture of
Tests
procedure for removal of bacterial endotoxins complies with
the following additional requirement. pH (2.4.24). 4.5 to 6.5, determined in a 10.0 per cent w/v solution.
Bacterial endotoxins (2.2.3). Not more than 0.20 Endotoxin Bacterial endotoxins (2.2.3). Not more than 0.20 Endotoxin
Unit per mg of cefotaxime. Unit per mg of cefotaxime.
Cefotaxime Sodium intended for use in the manufacture of Water (2.3.43). Not more than 6.0 per cent, determined on
parenteral preparations without a further appropriate 0.15 g.
additional requirement. Assay
Sterility (2.2.11). Complies with the test for sterility. Determine by liquid chromatography (2.4.14).
Storage. Store protected from moisture in tamper-evident Test solution. Determine the weight of the contents of
containers. 10 containers. Weigh accurately a quantity of the mixed
contents of the 10 containers dissolve in water and dilute to
Labelling. The label states whether or not the contents are obtain a solution containing 0.01 per cent w/v of cefotaxime.
Reference solution. A 0.01 per cent w/v solution of cefotaxime
sodium RS in water.
Cefotaxime Sodium Injection – a stainless steel column 30 cm x 3.9 mm, packed with
Cefotaxime Injection octadecylsilyl silica gel (3 to 10 µm),
– mobile phase: a solution prepared by dissolving 60 mg
Cefotaxime Sodium Injection is a sterile material consisting of of potassium dihydrogen phosphate and 1.2 g of
Cefotaxime Sodium with or without excipients. It is filled in disodium hydrogen phosphate in 1000 ml of water and
sealed containers. mixing with 120 ml of methanol,
The injection is constituted by dissolving the contents of the – flow rate. 1.5 ml per minute,
sealed container in the requisite amount of sterile Water for – spectrophotometer set at 254 nm,
Injections, immediately before use. – a 20 µl loop injector.
The constituted solution complies with the requirements for Inject the reference solution. The test is not valid unless the
Clarity of solution and Particulate matter stated under relative standard deviation for replicate injections is not more
Parenteral Preparations (Injections). than 2.0 per cent.
recommended by the manufacturer. Calculate the content of C16H17N5O7S2 in the injection.
Cefotaxime Sodium Injection contains a quantity of Cefotaxime Storage. Store protected from light at a temperature not
Sodium equivalent to not less than 90.0 per cent and not more exceeding 30°.
257
CEFTAZIDIME IP 2007
Ceftazidime – spectrophotometer set at 254 nm,

Inject the reference solution. Adjust the sensitivity of the
COO system so that the height of the principal peak in the
H2N
O chromatogram obtained is at least 50 per cent of the full scale
N O N N
S deflection of the recorder.
N S , 5H2O Inject the reference solution six times. The test is not valid
N H H H unless the relative standard deviation of the area of the principal
O peak in replicate injections is not more than 3.0 per cent.
H3C
COOH Inject alternately the test solution and the reference solution.
H3C Record the chromatograms and measure the areas of the
pyridine peaks.
C22H22N6O7S2,5H2O Mol. Wt. 636.6
Calculate the content of pyridine.
Ceftazidime is pentahydrate of the inner salt of (7R)-7-[(Z)-2-
(2-aminothiazol-yl)-2-(1-carboxy-1-methylethoxyimino)-
0.3 g by drying in an oven over phosphorus pentoxide at 60°
acetamido]-3-(1-pyridinomethyl)-3-cepham-4-carboxylate.
at a pressure not exceeding 0.7 kPa for 3 hours.
Ceftazidime contains not less than 95.0 per cent and not more Assay. Determine by liquid chromatography (2.4.14).
than 102.0 per cent of C22H22N6O7S2, calculated on the dried
basis. Test solution. Dissolve about 30 mg of the substance under
examination in 2.5 ml of phosphate buffer pH 7.0, dilute to
Description. A white to cream-coloured, crystalline powder. 25.0 ml with water and mix. Protect this solution from light.
Immediately before chromatography, dilute 5.0 ml of this
Identification
solution to 50.0 ml with water.
In the Assay, the principal peak in the chromatogram obtained Reference solution. Treat 30 mg ceftazidime RS in a similar
with the test solution corresponds to the peak in the manner.
Tests – a stainless steel column 15 cm x 4.6 mm, packed with
pH (2.4.24). 3.0 to 4.0, determined in a 0.5 per cent w/v solution. – mobile phase: a mixture of 100 ml of phosphate buffer
Pyridine. Not more than 0.4 per cent. pH 7.0 and 20 ml of acetonitrile diluted to 1000 ml with
water,
Determine by liquid chromatography (2.4.14). – flow rate. 2 ml per minute,
Test solution. Weigh accurately a quantity containing 0.5 g of – spectrophotometer set at 254 nm,
ceftazidime and dissolve in sufficient mixed phosphate buffer – a 10 µl loop injector.
pH 7.0 to produce 100.0 ml. Inject the reference solution. The test is not valid unless the
Reference solution. Weigh accurately about 200 mg of pyridine relative standard deviation for replicate injections is not more
and dissolve in sufficient water to produce 100.0 ml. than 2.0 per cent.
Immediately prior to chromatography add to 2.0 ml of the Inject alternately the test solution and the reference solution.
resulting solution sufficient mixed phosphate buffer pH 7.0
Calculate the content of C22H22N6O7S2.
to produce 200.0 ml and mix well.
Ceftazidime intended for use in the manufacture of parenteral
Chromatographic system preparations without a further appropriate procedure for
– a stainless steel column 25 cm x 4.6 mm, packed with the removal of bacterial endotoxins complies with the
octadecylsilyl silica gel (3 to 10 µm), following additional requirement.
w/v solution of ammonium dihydrogen phosphate Bacterial endotoxins (2.2.3). Not more than 0.10 Endotoxin
previously adjusted to pH 7.0 with dilute ammonia Unit per mg.
solution, 30 volumes of acetonitrile and 60 volumes of Ceftazidime intended for use in the manufacture of parenteral
water, preparations without a further appropriate sterilisation
– column temperature. 38° to 42°, procedure complies with the following additional
– flow rate. 1.6 ml per minute, requirement.
258
IP 2007 CEFTAZIDIME FOR INJECTION
Sterility (2.2.11). Complies with the test for sterility. Inject the reference solution six times. The test is not valid
Storage. Store protected from light and moisture. unless the relative standard deviation of the area of the principal
peak in replicate injections is not more than 3.0 per cent.
Measure the areas of the pyridine peaks.
Ceftazidime For Injection
Calculate the content of pyridine.
Ceftazidime for Injection is a sterile mixture of sterile
Ceftazidime and Sodium Carbonate. Sodium carbonate. Weigh accurately a quantity containing
about 50 mg of anhydrous ceftazidime and dissolve in
Ceftazidime for Injection contains not less than 90.0 per cent sufficient water to produce 100.0 ml. Dilute the resulting
and not more than 105.0 per cent of ceftazidime, C22H22N6O7S2, solution appropriately with water and determine by Method
calculated on the dried and sodium carbonate-free basis. A for flame photometry (2.4.4), measuring at 589 nm or by
Description. A white or almost white powder. Method A for atomic absorption spectrophotometry (2.4.2),
using sodium solution FP, suitably diluted with water for the
Identification reference solutions.
A. In the Assay, the principal peak in the chromatogram 1 g of Na is equivalent to 2.305 g of Na2CO3.
obtained with the test solution corresponds to the peak in the Bacterial endotoxins (2.2.3). Not more than 0.10 Endotoxin
chromatogram obtained with the reference solution. Unit per mg of ceftazidime.
B. Gives the reactions of sodium salts and reaction A of Sterility (2.2.11). Complies with the tests for sterility.
carbonates (2.3.1).
Tests determined on 0.3 g by drying at 25° for 4 hours at a pressure
not exceeding 0.7 kPa and continuing the drying by heating in
pH (2.4.24). 5.0 to 7.5, determined in a solution containing an oven at 100° for 3 hours at a pressure not exceeding 0.7
100 mg of anhydrous ceftazidime per ml. kPa.
Pyridine. Not more than 0.4 per cent. Assay. Determine by liquid chromatography (2.4.14).
Determine by liquid chromatography (2.4.14). Test solution. Weigh accurately a quantity containing about
Test solution. Weigh accurately a quantity containing about 50 mg of anhydrous ceftazidime dissolve in water and dilute
0.5 g of ceftazidime and dissolve in sufficient mixed phosphate to 50.0 ml with the same solvent. Protect this solution from
buffer pH 7.0 to produce 100.0 ml. light. Immediately before chromatography, dilute 5.0 ml to
50.0 ml with water.
Reference solution. Weigh accurately about 200 mg of pyridine
and dissolve in sufficient water to produce 100.0 ml. Reference solution. Dissolve about 29 mg ceftazidime RS in
Immediately prior to chromatography add to 2.0 ml of the 2.5 ml of mixed phosphate buffer pH 7.0 and dilute to 25.0 ml
resulting solution sufficient mixed phosphate buffer pH 7.0 with water. Protect this solution from light. Immediately before
to produce 200.0 ml and mix well, chromatography, dilute 5.0 ml to 50.0 ml with water.
Chromatographic system Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with – a stainless steel column 15 cm x 4.6 mm, packed with
octadecylsilyl silica gel (3 to 10 µm), octadecylsilyl silica gel (5 µm),
– mobile phase: a mixture of 10 volumes of a 2.88 per cent – mobile phase: a mixture of 100 ml of phosphate buffer
w/v solution of ammonium dihydrogen phosphate pH 7.0 and 20 ml of acetonitrile diluted to 1000 ml with
previously adjusted to pH 7.0 with dilute ammonia water,
solution, 30 volumes of acetonitrile and 60 volumes of – flow rate. 2 ml per minute,
water, – spectrophotometer set at 254 nm,
– flow rate. 1.6 ml per minute, – a 10 µl loop injector.
– spectrophotometer set at 254 nm, Inject the reference solution. The test is not valid unless the
– a 10 µl loop injector. relative standard deviation for replicate injections is not more
Inject the reference solution and adjust the sensitivity of the than 2.0 per cent.
system so that the height of the principal peak in the
chromatogram obtained is at least 50 per cent of the full-scale
deflection of the recorder. Calculate the content of C22H22N6O7S2.
259
CEFTAZIDIME INJECTION IP 2007
Storage. Store in sterile containers, sealed so as to exclude resulting solution sufficient mixed phosphate buffer pH 7.0
microorganisms, protected from moisture, at a temperature to produce 200.0 ml and mix well.
not exceeding 30°. Chromatographic system
Labelling. The label states the strength in terms of the – a stainless steel column 25 cm x 4.6 mm, packed with
equivalent amount of ceftazidime. octadecylsilyl silica gel (5 µm),
w/v solution of ammonium dihydrogen phosphate
previously adjusted to pH 7.0 with dilute ammonia
Ceftazidime Injection solution, 30 volumes of acetonitrile and 60 volumes of
Ceftazidime Injection is a sterile material consisting of water,
Ceftazidime for Injection with or without auxiliary substances. – flow rate. 1.6 ml per minute,
It is filled in a sealed container. – spectrophotometer set at 254 nm,
sealed container in the requisite amount of Water for Injections Inject the reference solution and adjust the sensitivity of the
immediately before use. system so that the height of the principal peak in the
chromatogram obtained is at least 50 per cent of the full-scale
The constituted solution complies with the requirements for deflection of the recorder.
Parenteral Preparations (Injections). Inject the reference solution six times. The test is not valid
unless the relative standard deviation of the area of the principal
Storage. The constituted solution should be used immediately peak in replicate injections is not more than 3.0 per cent.
recommended by the manufacturer. Inject alternately the test solution and the reference solution.
Measure the areas of the pyridine peaks.
Ceftazidime Injection contains not less than 90.0 per cent and
not more than 120.0 per cent of the stated amount of Calculate the content of pyridine.
ceftazidime, C22H22N6O7S2. Bacterial endotoxins (2.2.3). Not more than 0.10 Endotoxin
Description. A white or almost white crystalline powder. Unit per mg of ceftazidime.
The contents of the sealed container comply with the Sterility (2.2.11). Complies with the test for sterility.
requirements stated under Parenteral Preparations Loss on drying (2.4.19). Not more than 13.5 per cent,
(Powders for Injection) and with the following requirements. determined on 0.3 g by drying at 25° for 4 hours at a pressure
not exceeding 0.7 kPa and continuing the drying by heating in
Identification an oven at 100° for 3 hours at a pressure not exceeding 0.7
A. In the Assay, the principal peak in the chromatogram kPa.
obtained with the test solution corresponds to the peak in the Assay. Determine by liquid chromatography (2.4.14).
Test solution. Determine the weight of the contents of
B. Gives the reactions of sodium salts and reaction A of 10 containers. Weigh accurately a quantity of the mixed
carbonates (2.3.1). contents of the 10 containers containing about 50 mg of
ceftazidime, dissolve in water and dilute to 50.0 ml with the
Tests same solvent. Protect this solution from light. Immediately
pH (2.4.24). 5.0 to 7.5, determined in a solution containing before chromatography, dilute 5.0 ml to 50.0 ml with water.
100 mg of ceftazidime per ml. Reference solution. Dissolve about 29 mg ceftazidime RS in
Pyridine. Not more than 0.4 per cent. 2.5 ml of mixed phosphate buffer pH 7.0 and dilute to 25.0 ml
with water. Protect this solution from light. Immediately before
Determine by liquid chromatography (2.4.14). chromatography, dilute 5.0 ml to 50.0 ml with water.
Test solution. Weigh accurately a quantity containing about Chromatographic system
0.5 g of ceftazidime and dissolve in sufficient mixed phosphate – a stainless steel column 15 cm x 4.6 mm, packed with
buffer pH 7.0 to produce 100.0 ml. octadecylsilyl silica gel (5 µm),
Reference solution. Weigh accurately about 200 mg of pyridine – mobile phase: a mixture of 100 ml of phosphate buffer
and dissolve in sufficient water to produce 100.0 ml. pH 7.0 and 20 ml of acetonitrile diluted to 1000 ml with
Immediately prior to chromatography add to 2.0 ml of the water,
260
IP 2007 CEFTRIAXONE SODIUM
– flow rate. 2 ml per minute, more intensely coloured than reference solution BYS5 or YS5
– spectrophotometer set at 254 nm, (2.4.1).
– a 10 µl loop injector. pH (2.4.24). 6.0 to 8.0, determined in solution A.
Specific optical rotation (2.4.22). -155.0º to -170.0º, determined
in a 1.0 per cent w/v solution in water.
than 2.0 per cent.
(2.4.14).
Calculate the content of C22H22N6O7S2 in the injection. Test solution. Dissolve 30.0 mg of the substance under
Storage. Store in sterile containers, sealed so as to exclude examination in the mobile phase and dilute to 100.0 ml with the
microorganisms, protected from moisture, at a temperature mobile phase.
not exceeding 30 °.
Labelling. The label states the strength in terms of the ceftriaxone sodium RS in the mobile phase.
equivalent amount of ceftazidime.
w/v each of ceftriaxone sodium RS and ceftriaxone sodium
E- isomer RS in the mobile phase.
Ceftriaxone Sodium Reference solution (c). Dilute 1.0 ml of the reference solution
(a) to 100.0 ml with the mobile phase.
O
CH3 ONa
COONa N – a stainless steel column 25 cm x 4.6 mm, packed with
O O N
N N S N octadecylsilyl silica (5 µm),
H , 31/2 H20
N N CH3 – mobile phase: 2.0 g of tetradecylammonium bromide
S
H2N H H and 2.0 g of tetraheptylammonium bromide in a mixture
O
S of 440 ml of water, 55 ml of 0.067 M mixed phosphate
buffer solution pH 7.0, 5.0 ml of a buffer solution
C18H16N8Na2O7S3,3½H2O Mol. Wt. 662.0 prepared by dissolving 20.17 g of citric acid in 800 ml of
Ceftriaxone sodium is disodium (6R,7R)-7-[[(Z)-(2- water, adjusting the pH to 5.0 with strong sodium
aminothiazol-4-yl)(methoxyimino)acetyl]amino]-3-[[(2- hydroxide solution and diluting to 1000.0 ml with water,
methyl-6-oxido-5-oxo-2,5-dihydro-1,2,4-triazin-3-yl) and 500 ml of acetonitrile,
sulphanyl]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene- – flow rate. 1.5 ml per minute,
2-carboxylate. – spectrophotometer set at 254 nm,
Ceftriaxone sodium contains not less than 96.0 per cent and
not more than 102.0 per cent of C18H16N8Na2O7S3, calculated Inject the test solution and reference solutions (b) and (c).
on the anhydrous basis. Continue the chromatography for twice the retention time of
the ceftriaxone peak. The test is not valid unless the resolution
Description. A white or yellowish, crystalline powder, slightly
between the peaks due to ceftriaxone and ceftriaxone sodium
hygroscopic.
E-isomer is at least 3.0; the area of any peak other than the
Identification principal peak is not more than the area of the principal peak in
the chromatogram obtained with reference solution (c) (1 per
A. Determine by infrared absorption spectrophotometry (2.4.6). cent); the sum of the areas of all such peaks is not more than
Compare the spectrum with that obtained with ceftriaxone 4 times the area of the principal peak in the chromatogram
sodium RS or with the reference spectrum of ceftriaxone obtained with reference solution (c) (4 per cent). Ignore any
sodium. peak with an area 0.1 times the area of the principal peak in the
B. Gives reaction A of sodium salts (2.3.1). chromatogram obtained with reference solution (c) (0.1 per
cent).
Tests Water (2.3.43). 8.0 per cent to 11.0 per cent, determined on
Appearance of solution. Dissolve 2.4 g in 20 ml of carbon 0.1 g.
dioxide-free water (Solution A). Dilute 2 ml of solution A to 20 Assay. Determine by liquid chromatography (2.4.14) as
ml with water; the resulting solution is clear (2.4.1) and not described under Related substances.
261
CEFTRIAXONE INJECTION IP 2007
Inject alternately the test solution and reference solution (a). Tests
Calculate the content of C18H16N8Na2O7S3. Appearance of solution. A 1.2 per cent w/v solution in carbon
Ceftriaxone sodium intended for use in the manufacture of dioxide-free water is clear (2.4.1) and not more intensely
parenteral preparations without a further appropriate coloured than reference solution BYS5 or YS5 (2.4.1).
procedure for removal of bacterial endotoxins complies with pH (2.4.24). 6.0 to 8.0, determined in a 10.0 per cent w/v solution.
the following additional requirement.
Bacterial endotoxins (2.2.3): Not more than 0.20 Endotoxin (2.4.14).
Unit per mg of ceftriaxone sodium.
Test solution. Dissolve the substance under examination
Storage. Store protected from light and moisture. containing about 30 mg of ceftriaxone in 100 ml of the mobile
Labelling. The label states, where applicable, that the phase and filter.
substance is free from bacterial endotoxins. Reference solution (a). A 0.03 per cent w/v solution of
ceftriaxone sodium RS in the mobile phase.
w/v each of ceftriaxone sodium RS and ceftriaxone sodium
Ceftriaxone Injection E-isomer RS in the mobile phase.
Ceftriaxone Injection is a sterile material consisting of Reference solution (c). Dilute 1.0 ml of the reference solution
Ceftriaxone Sodium with or without excipients. It is filled in a (a) to 100.0 ml with the mobile phase.
sealed container.
The injection is constituted by dissolving the contents of the – a stainless steel column 25 cm × 4.6 mm, packed with
sealed container in the requisite amount of sterile Water for octadecylsilane bonded to porous silica (5 µm) (such as
Injections, immediately before use. Lichrosphere RP-18),
The constituted solution complies with the requirements for – mobile phase: dissolve 2 g of tetradecylammonium
Clarity of solution and Particulate matter stated under bromide and 2 g of tetraheptylammonium bromide in a
Parenteral Preparations (Injections). mixture of 440 ml of water, 55 ml of 0.067 M mixed
phosphate buffer pH 7.0, 5 ml of a buffer prepared by
Storage. The constituted solution should be used immediately dissolving 20.17 g of citric acid in 800 ml of water,
after preparation but, in any case, within the period adjusting the pH to 5.0 with 10 M sodium hydroxide
recommended by the manufacturer. and diluting to 1000 ml with water, and 500 ml of
Ceftriaxone Injection contains not less than 90.0 per cent and acetonitrile,
not more than 115.0 per cent of the stated amount of – flow rate. 1.5 ml per minute,
ceftriaxone, C18H18N8O7S3. – spectrophotometer set at 254 nm,
The contents of the sealed container comply with the resolution between the two principal peaks is at least 3.0.
(Powders for Injection) and with the following requirements. Inject the test solution and reference solution (c). Run
thechromatogram at least twice the retention time of the
Identification principal peak. In the chromatogram obtained with the test
solution the area of any secondary peak is not greater than
A. Determine by infrared absorption spectrophotometry (2.4.6). the area of the principal peak in the chromatogram obtained
Compare the spectrum with that obtained with ceftriaxone with reference solution (c) (1.0 per cent) and the sum of the
sodium RS or with the reference spectrum of ceftriaxone areas of all the secondary peaks is not greater than 5 times the
sodium. area of the principal peak in the chromatogram obtained with
reference solution (c) (5.0 per cent). Ignore any peak with an
area less than 0.1 times the area of the principal peak in the
obtained with the test solution
chromatogram obtained with reference solution (c) (0.1 per
corresponds to the peak in the chromatogram obtained with cent).
the reference solution (a).
Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit
C. It gives the reaction A of sodium salts (2.3.1). per mg of ceftriaxone.
262
IP 2007 CEFUROXIME AXETIL
Water (2.3.43). Not more than 11.0 per cent, determined on In the chromatogram obtained with the test solution, the ratio
0.1 g. of the peak due to cefuroxime axetil diastereoisomer A to the
Assay. Determine by liquid chromatography (2.4.14) as sum of the peaks due to cefuroxime axetil diastereoisomers A
described under Related substances. and B is between 0.48 and 0.55 by the normalisation procedure.
(2.4.14).
Calculate the content of C18H18N8O7S3 in the injection.
Use the chromatographic system, the test solution, and
Storage. Store protected from light at a temperature not reference solutions (a), (b), (c) and (d) described under Assay.
exceeding 30º.
The percentage sum of the pair of peaks corresponding to the
Labelling. The label on the sealed container states the quantity E-isomers located by comparison with the chromatogram
of Ceftriaxone Sodium contained in it in terms of the equivalent obtained with reference solution (c) is not more than 1.0 per
amount of ceftriaxone. cent, the percentage sum of the pair of peaks corresponding
to the D 3 -isomers located by comparison with the
chromatogram obtained with reference solution (b) is not
greater than 1.5 per cent and the area of any other secondary
Cefuroxime Axetil peak is not more than 0.5 per cent and the sum of all the
secondary peaks found is not more than 3.0 per cent.
H 3C O CH3
Acetone (5.4). Not more than 1.1 per cent.
O O O Water (2.3.43). Not more than 1.5 per cent, determined on
O
H3CO O 0.4 g.
N H N O NH2 Assay. Determine by liquid chromatography (2.4.14).
N
S NOTE – Prepare the solutions immediately before use.
H H
O O Test solution. Dissolve 10.0 mg of the substance under
examination in the mobile phase and dilute to 50.0 ml with the
C20H22N4O10S Mol. Wt. 510.5 mobile phase.
Cefuroxime Axetil is a mixture of the 2 diastereoisomers of Reference solution (a). Dilute 1.0 ml of the test solution to
(1RS)-1-(acetyloxy)ethyl (6R,7R)-3-[(carbamoyloxy)methyl]- 100.0 ml with the mobile phase.
7-[[(Z)-2-(furan-2-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-
5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate. Reference solution (b). Warm 5 ml of the test solution, to 60º
for one hour to generate the D3-isomers.
Cefuroxime Axetil contains not less than 79.8 per cent and not
more than 84.8 per cent of cefuroxime, C16H16N4O8S, calculated Reference solution (c) Expose 5 ml of the test solution to
on the anhydrous and acetone-free basis. ultraviolet light at 254 nm for 24 hours to generate E-isomers.
Description. A white or almost white powder. Reference solution (d). A 0.02 per cent w/v solution of
cefuroxime axetil RS in the mobile phase.
A. Determine by infrared absorption spectrophotometry (2.4.6). – a stainless steel column 25 cm × 4.6 mm, packed with
Compare the spectrum with that obtained with cefuroxime trimethylsilyl silica gel (5 µm),
axetil RS or with the reference spectrum of cefuroxime axetil. – mobile phase: a mixture of 38 volumes of methanol and
62 volumes of a 2.3 per cent solution of ammonium
B. In the Assay, the principal peaks in the chromatogram dihydrogen phosphate,
obtained with the test solution corresponds to the peak in the – flow rate. 1 ml per minute,
chromatogram obtained with reference solution (d). – spectrophotometer set at 278 nm,
Tests
Inject reference solution (d). The test is not valid unless the
Diastereoisomer ratio. Determine by liquid chromatography relative standard deviation for replicate injections is not more
(2.4.14). than 2.0 per cent.
Use the chromatographic system, the test solution, and Inject the test solution, and reference solutions (a), (b) and
reference solutions (a), (b), (c) and (d) described under Assay. (c). The retention times relative to cefuroxime axetil
263
CEFUROXIME AXETIL TABLETS IP 2007
diastereoisomer A (second peak) are approximately 0.9 for Related substances. Determine by liquid chromatography
cefuroxime axetil diastereoisomer B, 1.2 for the cefuroxime axetil (2.4.14).
D3-isomers and 1.7 and 2.1 for the E-isomers. The test is not Use the chromatographic system, the test solution and
valid unless in the chromatogram obtained with reference reference solutions (a), (b), and (c) described under Assay.
solution (d), the resolution between the peaks corresponding
to cefuroxime axetil diastereoisomers A and B is at least 1.5. In In the chromatogram obtained with the test solution the sum
the chromatogram obtained with reference solution (b), the of the areas of the pair of peaks corresponding to the E-isomers
resolution between the peaks corresponding to cefuroxime in the chromatogram obtained with reference solution (b) is
axetil diastereoisomer A and cefuroxime axetil D3-isomer is at not more than 1.5 per cent by normalisation, the sum of the
least 1.5. areas of any peaks corresponding to the D3-isomers in the
chromatogram obtained with reference solution (a) is not more
Calculate the content of C16H16N4O8S as the sum of areas of
than 2.0 per cent by normalisation and the area of any other
the two diastereoisomer peaks.
secondary peak is not more than 1.0 per cent by normalisation.
1 mg of C20H22N4O10S is equivalent to 0.8313 mg of C16H16N4O8S.
Storage. Store protected from light and moisture. Assay. Determine by liquid chromatography (2.4.14).
NOTE – Prepare the solutions immediately before use.
Test solution. Disperse 10 tablets in 0.2 M ammonium
Cefuroxime Axetil Tablets dihydrogen orthophosphate with the pH previously adjusted
Cefuroxime Axetil Tablets contain Cefuroxime Axetil. They may to 2.4 with orthophosphoric acid, using 10 ml per g of the
be coated. stated content of cefuroxime. Immediately add sufficient
Cefuroxime Axetil Tablets contain not less than 90.0 per cent methanol to produce a solution containing the equivalent of
and not more than 110.0 per cent of the stated amount of 0.5 per cent w/v of cefuroxime and shake vigorously. Filter
cefuroxime, C16H16N4O8S. and dilute a quantity of the filtrate with sufficient of the mobile
phase to produce a solution containing 0.025 per cent w/v of
Identification cefuroxime.
Reference solution (a). Warm a quantity of the test solution
A. Extract a quantity of the powdered tablets containing 0.1 g
at 60º for one hour or until sufficient impurities (D3-isomers)
of cefuroxime with 5 ml of dichloromethane, filter and
have been generated.
evaporate the filtrate to dryness.
Reference solution (b). Expose a quantity of the test solution
to ultraviolet light at 254 nm for 24 hours or until sufficient
Compare the spectrum with that obtained with cefuroxime
impurities (E-isomers) have been generated.
axetil RS or with the reference spectrum of cefuroxime axetil.
B. In the Assay, the principal peaks in the chromatogram
cefuroxime axetil RS in the mobile phase.
chromatogram obtained with reference solution (c). Chromatographic system
Tests particles of silica (5 µm) the surface of which has been
modified by chemically-bonded trimethylsilyl groups
(such as Hypersil SAS),
Apparatus. No. 1 – mobile phase: a mixture of 38 volumes of methanol and
Medium. 900 ml of 0.1 M hydrochloric acid 62 volumes of 0.2 M ammonium dihydrogen
Speed and time. 50 rpm and 45 minutes. orthophosphate, adjusted, if necessary, so that the
resolution between the peaks corresponding to the
Withdraw a suitable volume of the medium and filter promptly.
cefuroxime axetil diastereoisomers A and B in reference
Dilute the filtrate, if necessary, with the dissolution medium.
solution (c) and between the peaks corresponding to
Measure the absorbance of the resulting solution at the
cefuroxime axetil diastereoisomer A and the cefuroxime
maximum at about 278 nm (2.4.7). Calculate the content of
axetil D3-isomer in reference solution (a) is in each case
C16H16N4O8S in the medium from the absorbance obtained from
not less than 1.5,
a solution of known concentration of cefuroxime axetil RS.
D. Not less than 70 per cent of the stated amount of – spectrophotometer set at 278 nm,
C16H16N4O8S. – a 20 µl loop injector.
264
IP 2007 CEFUROXIME INJECTION
Inject reference solution (c). The test is not valid unless the in sufficient water to produce 25.0 ml. Immediately transfer
relative standard deviation for replicate injections is not more 5.0 ml of the resulting solution to a 100-ml volumetric flask,
than 2.0 per cent. add 20.0 ml of a 0.15 per cent w/v solution of orcinol (internal
Inject the test solution, reference solutions (a), (b) and (c). standard) in water, dilute to volume with water and mix.
The retention time relative to cefuroxime axetil diastereoisomer
Reference solution. Treat a quantity of cefuroxime sodium RS
A are approximately 0.9 for cefuroxime axetil
equivalent to 25 mg of cefuroxime in a similar manner.
diastereoisomer B, 1.2 for the cefuroxime axetil D3-isomers and
1.7 and 2.1 for the E-isomers.
Calculate the content of C16H16N4O8S as the sum of the areas hexylsilane chemically bonded to totally porous silica
of the two peaks corresponding to diastereoisomers A and B. particles (5 µm),
1 mg of C20H22N4O10S is equivalent to 0.8313 mg of C16H16N4O8S. – mobile phase: a mixture of 100 volumes of acetate buffer
pH 3.4 and 10 volumes of acetonitrile,
Labelling. The quantity of active ingredient is stated in terms – flow rate. 2 ml per minute,
of the equivalent amount of cefuroxime. – spectrophotometer set at 254 nm,
Cefuroxime Sodium relative standard deviation for replicate injections is not more
than 2.0 per cent.
COONa O Inject alternately the test solution and the reference solution.
O
O N O NH2 Calculate the content of C16H16N4O8S.
O
Cefuroxime Sodium intended for use in the manufacture of
N S parenteral preparations complies with the following
N H H H
H3CO additional requirements.
C16H15N4NaO8S Mol. Wt. 446.4 Unit per mg of cefuroxime.
Cefuroxime Sodium is sodium (7R)-3-carbamoyloxymethyl-7- Sterility (2.2.11). Complies with the test for sterility, using the
[(Z)-furan-2-yl-2-methoxyiminoacetamido]-3-cephem-4- membrane filtration method.
carboxylate.
Storage. Store protected from moisture. If it is intended for
Cefuroxime Sodium contains not less than 90.0 per cent and use in the manufacture of parenteral preparations, it should
not more than 105.0 per cent of cefuroxime, C16H16N4O8S, be sterile and sealed so as to exclude micro-organisms.
calculated on the anhydrous basis.
Description. A white or faintly yellow powder. intended for use in the manufacture of injectable preparations.
Identification
obtained with the test solution corresponds to the peak in the Cefuroxime Injection
Cefuroxime Sodium Injection
B. Gives the reactions of sodium salts (2.3.1).
Cefuroxime Injection is a sterile material consisting of
Tests Cefuroxime Sodium, with or without auxiliary substances. It
is filled in a sealed container.
The injection is constituted by dissolving the contents of a
Water (2.3.43). Not more than 3.5 per cent, determined on sealed container in the requisite amount of Water for Injections
0.15 g. immediately before use.
Assay. Determine by liquid chromatography (2.4.14). The constituted solution complies with the requirements for
Test solution. Weigh accurately a quantity of the substance Clarity of solution and Particulate matter stated under
under examination containing 25 mg of cefuroxime and dissolve Parenteral Preparations (Injections).
265
CELLULOSE ACETATE PHTHALATE IP 2007
after preparation but, in any case, within the period Calculate the content of C16H16N4O8S in the injection.
Storage.Store in tightly-closed containers protected from
Cefuroxime Injection contains a quantity of Cefuroxime Sodium moisture at a temperature not exceeding 30°.
120.0 per cent of the stated amount of cefuroxime, C16H16N4O8S. Labelling. The label on the sealed container states the quantity
of Cefuroxime Sodium contained in it in terms of the equivalent
Description. A white or faintly yellow powder. amount of cefuroxime.
Cellulose Acetate Phthalate
Identification Cellacephate; Cellacefate
A. In the Assay, the principal peak in the chromatogram Cellulose Acetate Phthalate is a cellulose, some of the
obtained with the test solution corresponds to the peak in the hydroxyl groups of which are esterified by acetyl groups
chromatogram obtained with the reference solution. and others by hydrogen phthaloyl groups.
B. Gives the reactions of sodium salts (2.3.1). Cellulose Acetate Phthalate contains not less than 17.0 per
cent and not more than 26.0 per cent of acetyl groups, C2H3O
Tests and not less than 30.0 per cent and not more than 40.0 per cent
of hydrogen phthaloyl groups, C8H5O3 both calculated on the
pH (2.4.24). 6.0 to 8.5, determined in a 10.0 per cent w/v solution. dried, acid-free basis.
Bacterial endotoxins (2.2.3). Not more than 0.1 Endotoxin Unit Description. A white, free-flowing powder or colourless flakes;
per mg of cefuroxime. odourless or with a faint odour of acetic acid; hygroscopic.
0.15 g. Identification
Assay. Determine by liquid chromatography (2.4.14). A. To about 10 mg add 1 ml of ethanol (95 per cent) and 1 ml
of sulphuric acid and warm; ethyl acetate, recognisable by its
Test solution. Determine the weight of the contents of
characteristic odour, is evolved.
10 containers. Weigh accurately a quantity of the mixed
contents of the 10 containers containing about 25 mg of B. To about 10 mg contained in a small test-tube add 10 mg of
cefuroxime and dissolve in sufficient water to produce resorcinol, mix, add 0.5 ml of sulphuric acid and heat in a
25.0 ml. Immediately transfer 5.0 ml of the resulting solution to liquid paraffin bath at 160° for 3 minutes. Cool and pour the
a 100-ml volumetric flask, add 20.0 ml of a 0.15 per cent w/v solution into a mixture of 25 ml of sodium hydroxide solution
solution of orcinol (internal standard) in water, dilute to volume and 200 ml of water; the solution shows a vivid green
with water and mix. fluorescence.
Reference solution. Treat a quantity of cefuroxime sodium RS
Tests
equivalent to 25 mg of cefuroxime in a similar manner.
Viscosity (2.4.28). 50 mm²s–1 to 90 mm²s–1, determined in the
following manner. Weigh accurately about 15 g, previously
dried at 105° for 2 hours, and dissolve in 85 g of a mixture of
hexylsilane chemically bonded to totally porous silica
249 parts of dry acetone and 1 part of water. Determine at 25°
particles (5 µm),
the viscosity of the resulting solution by Method A, using a
– mobile phase: a mixture of 100 volumes of acetate buffer
size D viscometer.
pH 3.4 and 10 volumes of acetonitrile,
– flow rate. 2 ml per minute, Appearance of a film. Dissolve 3.0 g in 17 ml of acetone with
– spectrophotometer set at 254 nm, a water content of 0.35 to 0.45 per cent w/w. Allow 1 ml of this
– a 10 µl loop injector. solution to flow over a glass plate and dry; a thin, colourless,
Inject the reference solution. The test is not valid unless the transparent and glossy film is produced.
relative standard deviation for replicate injections is not more Free acid. Not more than 3.0 per cent, calculated as phthalic
than 2.0 per cent. acid, C8H6O4, on the anhydrous basis and determined in the
266
IP 2007 CEPHALEXIN
following manner. Weigh accurately 1.0 g, in fine powder, shake Cephalexin

for 5 minutes with 100 ml of boiled water and filter. Wash the
flask and the filter with two quantities, each of 10 ml, of water.
Combine the filtrate and washings, add 5 drops of COOH
phenolphthalein solution and titrate with 0.1 M sodium O CH3
hydroxide until a faint pink colour is obtained. O N , H2O
1 ml of 0.1 M sodium hydroxide is equivalent to 0.0083 g of N S
phthalic acid. NH2 H H H
Heavy metals (2.3.13). 1.0 g complies with limit test for heavy
metals, Method B (20 ppm). C16H17N3O4S,H2O Mol. Wt. 365.4
Sulphated ash (2.3.18). Not more than 0.1 per cent. Cephalexin is (7R)-3-methyl-7-(α-D-phenylglycylamino)-3-
cephem-4-carboxylic acid monohydrate.
Water (2.3.43). Not more than 5.0 per cent, using 0.5 g dissolved
in 20 ml of a mixture of equal volumes of anhydrous methanol Cephalexin contains not less than 95.0 per cent and not more
and chloroform. than 101.0 per cent of C16H 17N3O4S, calculated on the
anhydrous basis.
Assay. For acetyl groups — Weigh accurately about 0.1 g
and heat on a water-bath for 30 minutes with 25.0 ml of 0.1 M
odour, characteristic.
sodium hydroxide under reflux. Cool, add 5 drops of
phenolphthalein solution and titrate with 0.1 M hydrochloric Identification
acid until the colour is discharged. Carry out a blank titration.
Calculate the acetyl groups, C2H3O, from the expression A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with cephalexin RS
0.43c/w - (0.578p + 0.518s), or with the reference spectrum of cephalexin.
where, c = volume, in ml, of 0.1 M hydrochloric acid B. In the Assay, the principal peak in the chromatogram
consumed obtained with the test solution corresponds to the peak in the
w = weight, in g, of the sample, calculated with
reference to anhydrous substance Tests
p = percentage of hydrogen phthaloyl groups as
determined in Assay for hydrogen phthaloyl
groups Specific optical rotation (2.4.22). +149° to +158°, determined
in a 0.5 per cent w/v solution in phthalate buffer solution pH
s = percentage of free acid.
4.4 and in a 2-dm tube.
For hydrogen phthaloyl groups - Weigh accurately about Light absorption (2.4.7). Dissolve 50 mg in sufficient water to
0.4 g (calculated on the anhydrous basis) and dissolve without produce 100.0 ml. Absorbance of the solution at about
heating in 20 ml of 2-methoxyethanol, previously neutralised 330 nm, not greater than 0.05 (2.4.7). Dilute 2 ml to 50.0 ml.
in the presence of 5 drops of phenolphthalein solution. Titrate When examined between 230 nm and 360 nm the solution
with 0.1 M sodium hydroxide until a faint pink colour is shows an absorption maximum at about 262 nm; absorbance
produced. Calculate the hydrogen phthaloyl groups, C8H5O3, at about 262 nm, 0.44 to 0.49.
from the expression
1.49b/w - 1.795s (2.4.14).
where, b = volume, in ml, of 0.1 M sodium hydroxide
examination in mobile phase A and dilute to 50 ml with the
consumed
same solvent.
w = weight, in g, of the sample, calculated with
Reference solution (a). Dissolve 10 mg of D-phenylglycine in
reference to the anhydrous substance
mobile phase A and dilute to 10 ml with the same solvent.
s = percentage of free acid.
Reference solution (b). Dissolve 10 mg of 7-
Storage. Store protected from moisture at a temperature aminodesacetoxycephalosporanic acid RS in phosphate
between 8° and 15°. buffer solution pH 7.0 and dilute to 10 ml with the same solvent.
267
CEPHALEXIN CAPSULES IP 2007
Reference solution (c). Dilute 1 ml of reference solution (a) chromatogram obtained with reference solution (e) (0.05 per
and 1 ml of reference solution (b) to 100 ml with mobile cent).
phase A. Sulphated ash (2.3.18). Not more than 0.2 per cent.
Reference solution (d). Dissolve 10 mg of dimethylformamide
and 10 mg of dimethylacetamide in mobile phase A and dilute
to 10 ml with the same solvent. Dilute 1 ml to 100 ml with Assay. Determine by liquid chromatography (2.4.14).
mobile phase A. Test solution. Dissolve 50 mg of the substance under
Reference solution (e). Dilute 1 ml of reference solution (c) to examination in water and dilute to 100.0 ml with the same
20 ml with mobile phase A. solvent.
Reference solution (f). Dissolve 10 mg of cefotaxime sodium Reference solution (a). Dissolve 50 mg of cephalexin
RS in mobile phase A and dilute to 10 ml with the same solvent. monohydrate RS in water and dilute to 100.0 ml with the same
To 1 ml of the solution add 1 ml of the test solution and dilute solvent.
to 100 ml with mobile phase A. Reference solution (b). Dissolve 10 mg of cephradine RS in
Chromatographic system 20 ml of reference solution (a) and dilute to 100 ml with water.
octadecylsilyl silica gel (5 µm), – a stainless steel column 25 cm x 4.6 mm, packed with
– mobile phase: A. a mixture of phosphate buffer solution octadecylsilyl silica gel (5 µm),
pH 5.0, – mobile phase: a mixture of 2 volumes of methanol,
B. methanol, 5 volumes of acetonitrile, 10 volumes of a 13.6 g per
– flow rate. 1.5 ml per minute, litre solution of potassium dihydrogen phosphate and
– a linear gradient programme using the conditions given 83 volumes of water,
below, – flow rate. 1.5 ml per minute,
– spectrophotometer set at 220 nm, – spectrophotometer set at 254 nm,
– a 20 µl loop injector. – a 20 µl loop injector.
Inject reference solution (b). In the chromatogram obtained,
the resolution between the peaks due to cephalexin and
0–1 98 2 cephradine is not less than 4.0.
1 – 20 98 → 70 2 → 30 Inject alternately the test solution and reference solution (a).
20 – 23 70 → 98 30 → 2 Calculate the content of C16H17N3O4S.
23 – 30 98 2
Inject reference solutions (c) and (f). In the chromatogram exceeding 30°.
obtained with reference solution (c) the resolution between
the peaks due to D-phenylglycine and 7-aminodesacetoxy-
cephalosporanic acid is not less than 2.0 and in the
chromatogram obtained with reference solution (f) the Cephalexin Capsules
resolution between the peaks due to cephalexin and cefotaxime
is not less than 1.5. Cephalexin Capsules contain not less than 90.0 per cent and
Inject the test solution and reference solutions (c) (d) and (e). anhydrous cephalexin, C16H17N3O4S.
In the chromatogram obtained with the test solution any peak
corresponding to D-phenylglycine is not more than the area Identification
of the second peak in the chromatogram obtained with
reference solution (c) (1.0 per cent). Ignore any peaks due to Shake a quantity of the contents of the capsules containing
dimethylformamide and dimethylacetamide The area of any 0.5 g of anhydrous cephalexin with 1 ml of water and 1.4 ml of
other secondary peak is not more than the area of the first 1 M hydrochloric acid, filter and wash the filter with 1 ml of
peak in the chromatogram obtained with reference solution water. Add slowly to the filtrate a saturated solution of sodium
(c) (1.0 per cent). The sum of the secondary peaks is not more acetate until precipitation occurs. Add 5 ml of methanol, filter
than the three times the area of the first peak in the and wash the precipitate with two quantities, each of 1 ml, of
chromatogram obtained with reference solution (c). Ignore methanol. The residue after drying at a pressure not exceeding
any peak with an area less than the second peak in the 0.7 kPa complies with the following tests.
268
IP 2007 CEPHALEXIN CAPSULES
A. Determine by infrared absorption spectrophotometry (2.4.6). Dissolution (2.5.2).

Compare the spectrum with that obtained with cephalexin RS Apparatus. No 1
or with the reference spectrum of cephalexin.
Medium. 900 ml of water freshly prepared by distillation.
B. In the Assay, the principal peak in the chromatogram Speed and time. 100 rpm and 45 minutes.
Withdraw a suitable volume of the medium and filter promptly
through a membrane filter disc with an average pore diameter
not greater than 0.8 µm. Reject the first few ml of the filtrate
Tests and dilute a suitable volume of the filtrate with water. Measure
the absorbance of the resulting solution at the maximum at
about 261 nm (2.4.7). Calculate the content of C16H17N3O4S
(2.4.17), coating the plate with silica gel HF (such as Analtech
plates). Impregnate the plate by development with a 5 per cent
v/v solution of n-tetradecane in hexane. Allow the solvent to D. Not less than 75 per cent of the stated amount of
evaporate and carry out the chromatography in the same C16H17N3O4S.
direction as the impregnation. Other tests. Complies with the tests stated under Capsules.
Mobile phase. A mixture of 3 volumes of acetone, 80 volumes Water (2.3.43). Not more than 10.0 per cent, determined on
of a 7.2 per cent w/v solution of disodium hydrogen 0.3 g of the contents of the capsules.
orthophosphate and 120 volumes of a 2.1 per cent w/v solution Assay. Determine by liquid chromatography (2.4.14).
of citric acid.
Test solution. Shake a quantity of the powdered mixed contents
Test solution. Shake a quantity of the contents of the capsules of 20 capsules containing about 0.25 g of anhydrous
containing about 0.25 g of anhydrous cephalexin with 10 ml of cephalexin with 100.0 ml of water for 30 minutes, add sufficient
2 M hydrochloric acid, filter and use the filtrate. of water to produce 250.0 ml and filter. Dilute 25.0 ml of the
Reference solution (a). Dilute 1 volume of the test solution to filtrate to 50.0 ml with water.
100 volumes with 2 M hydrochloric acid. Reference solution (a). A 0.05 per cent w/v solution of
cephalexin RS in water.
Reference solution (b). A 0.025 per cent w/v solution of 7-
aminodesacetoxycephalosporanic acid RS in 2 M Reference solution (b). A solution containing 0.01 per cent
hydrochloric acid. w/v each of cephalexin RS and cephradine RS in water.
Reference solution (c). A 0.025 per cent w/v solution of DL- Chromatographic system
phenylglycine in 2 M hydrochloric acid. – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilyl silica gel (5 µm) (such as Nucleosil C 18),
Reference solution (d). A solution containing 2.5 per cent – mobile phase: a mixture of 2 volumes of methanol,
w/v of cephalexin RS and 0.025 per cent w/v each of 7- 5 volumes of acetonitrile, 10 volumes of a 13.6 g per
aminodesacetoxycephalosporanic acid RS and DL- litre solution of potassium dihydrogen phosphate and
phenylglycine in 2 M hydrochloric acid. 83 volumes of water,
Apply to the plate 5 µl of each of the following solutions.
After development, dry the plate at 90° for 3 minutes, spray
the hot plate with a 0.1 per cent w/v solution of ninhydrin in
the mobile phase, heat the plate at 90° for 15 minutes and Inject reference solution (b). Adjust the sensitivity so that the
allow to cool. In the chromatogram obtained with the test height of the peaks in the chromatogram obtained is at least
solution any spot corresponding to 7-aminodesacetoxy- half the full-scale deflection on the recorder. The test is not
cephalosporanic acid is not more intense than the spot in the valid unless the resolution between the peaks corresponding
chromatogram obtained with reference solution (b) (1 per to cephalexin and cephradine is at least 4.0.
cent), any spot corresponding to DL-phenylglycine is not Inject reference solution (a) six times. The relative standard
more intense than the spot in the chromatogram obtained deviation is not more than 1.0 per cent.
with reference solution (c) (1 per cent) and any other secondary Inject alternately the test solution and reference solution (a).
spot is not more intense than the spot in the chromatogram
obtained with reference solution (a) (1 per cent). The test is Calculate the content of C16H17N3O4S in the capsules.
not valid unless the chromatogram obtained with reference Storage. Store protected from moisture at a temperature not
solution (d) shows three clearly separated spots. exceeding 30°.
269
CEPHALEXIN ORAL SUSPENSION IP 2007
Labelling. The label states the strength in terms of the Chromatographic system
equivalent amount of anhydrous cephalexin. – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilyl silica gel (5 µm) (such as Nucleosil C 18),
– mobile phase: a mixture of 2 volumes of methanol,
5 volumes of acetonitrile, 10 volumes of a 13.6 g per
Cephalexin Oral Suspension litre solution of potassium dihydrogen phosphate and
83 volumes of water,
Cephalexin Dry Syrup; Cephalexin Mixture – flow rate. 1.5 ml per minute,
Cephalexin Oral Suspension is a mixture of Cephalexin with – spectrophotometer set at 254 nm,
buffering agents and other excipients. It contains a suitable – a 20 µl loop injector.
flavouring agent. It is filled in a sealed container. Inject reference solution (b).Adjust the sensitivity so that the
The suspension is constituted by dispersing the contents of height of the peaks in the chromatogram obtained is at least
the sealed container in the specified volume of water just half the full-scale deflection on the recorder. The test is not
before use. valid unless the resolution between the peaks corresponding
to cephalexin and cephradine is at least 4.0.
Cephalexin Oral Suspension contains not less than 90.0 per
cent and not more than 120.0 per cent of the stated amount of Inject reference solution (a) six times. The relative standard
C16H17N3O4S. deviation is not more than 1.0 per cent.
When stored at the temperature and for the period stated on Inject alternately the test solution and reference solution (a).
the label during which the constituted suspension may be Determine the weight per ml (2.4.29) of the suspension and
expected to be satisfactory for use, it contains not less than calculate the content of C16H17N3O4S, weight in volume.
80.0 per cent of the stated amount of cephalexin, C16H17N3O4S.
Repeat the procedure using a portion of the suspension that
Identification has been stored at the temperature and for the period stated
on the label during which it may be expected to be satisfactory
A. In the Assay, the principal peak in the chromatogram for use.
Storage. Store protected from moisture, at a temperature not
chromatogram obtained with the reference solution (a).
exceeding 30°.
B. Weigh a quantity containing 0.1 g of anhydrous cephalexin,
shake with 20 ml of methanol, filter and evaporate the filtrate
equivalent amount of anhydrous cephalexin.
to dryness using a rotary evaporator. Dissolve the residue in
the minimum volume of a 1 per cent v/v solution of glacial
acetic acid, decolorise if necessary by the addition of sufficient
decolorising charcoal, shake and filter. To 0.25 ml of the
resulting solution add 0.1 ml of a 1 per cent w/v solution of Cephalexin Tablets
cupric sulphate and 0.05 ml of 2 M sodium hydroxide; an
olive-green colour is produced. Cephalexin Tablets contain not less than 90.0 per cent and not
The constituted suspension complies with the tests stated cephalexin, C16H17N3O4S. The tablets may be coated.
under Oral liquids and with the following tests.
Assay. Determine by liquid chromatography (2.4.14). Identification
Test solution. Shake an accurately weighed quantity of the Remove any coating. Shake a quantity of the powdered tablet
oral suspension containing about 0.25 g of anhydrous cores containing 0.5 g of anhydrous cephalexin with 1 ml of
cephalexin with 100.0 ml of water for 30 minutes, add sufficient water and 1.4 ml of 1 M hydrochloric acid, add 0.1 g of
of water to produce 250.0 ml and filter. Dilute 25.0 ml of the decolorising charcoal, shake, filter and wash the filter with
filtrate to 50.0 ml with water. 1 ml of water. Add slowly to the filtrate a saturated solution of
Reference solution (a). A 0.05 per cent w/v solution of sodium acetate until precipitation occurs. Add 5 ml of
cephalexin RS in water. methanol, filter and wash the precipitate with two quantities,
each of 1 ml, of methanol. The residue, after drying at a
Reference solution (b). A solution containing 0.01 per cent pressure not exceeding 0.7 kPa, complies with the following
w/v each of cephalexin RS and cephradine RS in water. test.
270
IP 2007 CEPHALEXIN TABLETS
A. Determine by infrared absorption spectrophotometry (2.4.6). Medium. 900 ml of water freshly prepared by distillation
Compare the spectrum with that obtained with cephalexin RS Speed and time. 100 rpm and 45 minutes.
or with the reference spectrum of cephalexin.
B. In the Assay, the principal peak in the chromatogram through a membrane filter disc with an average pore diameter
obtained with the test solution corresponds to the peak in the not greater than 1.0 µm. Reject the first few ml of the filtrate
chromatogram obtained with the reference solution (a). and dilute a suitable volume of the filtrate with water. Measure
Tests about 261 nm (2.4.7). Calculate the content of C16H17N3O4S
Related substances. Determine by thin-layer chromatography taking 235 as the specific absorbance at 261 nm.
(2.4.17), coating the plate with silica gel HF (such as Analtech D. Not less than 75 per cent of the stated amount of
plates). Impregnate the plate by development with a 5 per cent C16H17N3O4S.
v/v solution of n-tetradecane in hexane. Allow the solvent to
evaporate and carry out the chromatography in the same Tests
direction as the impregnation.
Mobile phase. A mixture of 3 volumes of acetone, 80 volumes
of a 7.2 per cent w/v solution of disodium hydrogen Assay. Determine by liquid chromatography (2.4.14).
orthophosphate and 120 volumes of a 2.1 per cent w/v solution Test solution. Weigh and powder 20 tablets. Shake a quantity
of citric acid. of the powdered tablets containing about 0.25 g of anhydrous
Test solution. Shake a quantity of the powered tablets cephalexin with 100.0 ml of water for 30 minutes, add sufficient
containing 0.25 g of anhydrous cephalexin with 10 ml of 2 M of water to produce 250.0 ml and filter. Dilute 25.0 ml of the
hydrochloric acid, filter and use the filtrate. filtrate to 50.0 ml with water.
Reference solution (a). Dilute 1 volume of the solution to Reference solution (a). A 0.05 per cent w/v solution of
100 volumes with 2 M hydrochloric acid. cephalexin RS in water.
Reference solution (b). A 0.025 per cent w/v solution of 7- Reference solution (b). A solution containing 0.01 per cent
aminodesacetoxycephalosporanic acid RS in 2 M w/v each of cephalexin RS and cephradine RS in water.
hydrochloric acid. Chromatographic system
Reference solution (c). A 0.025 per cent w/v solution of DL- – a stainless steel column 25 cm x 4.6 mm, packed with
phenylglycine in 2 M hydrochloric acid. octadecylsilyl silica gel (5 µm) (such as Nucleosil C 18),
– mobile phase: a mixture of 2 volumes of methanol,
5 volumes of acetonitrile, 10 volumes of a 13.6 g per
w/v of cephalexin RS and 0.025 per cent w/v each of 7-
litre solution of potassium dihydrogen phosphate and
aminodesacetoxycephalosporanic acid RS and DL -
83 volumes of water,
phenylglycine in 2 M hydrochloric acid.
Apply separately to the plate 5 µl of each solution. After – spectrophotometer set at 254 nm,
development, dry the plate at 90° for 3 minutes, spray the hot – a 20 µl loop injector.
plate with a 0.1 per cent w/v solution of ninhydrin in the
Inject reference solution (b). Adjust the sensitivity so that the
mobile phase, heat the plate at 90° for 15 minutes and allow to
height of the peaks in the chromatogram obtained is at least
cool. In the chromatogram obtained with the test solution any
half the full-scale deflection on the recorder. The test is not
spot corresponding to 7-aminodesacetoxycephalosporanic
valid unless the resolution between the peaks corresponding
acid is not more intense than the spot in the chromatogram
to cephalexin and cephradine is at least 4.0.
obtained with reference solution (b) (1 per cent), any spot
corresponding to DL-phenylglycine is not more intense than Inject reference solution (a) six times. The relative standard
the spot in the chromatogram obtained with reference solution deviation is not more than 1.0 per cent.
(c) (1 per cent) and any other secondary spot is not more Inject alternately the test solution and reference solution (a).
reference solution (a) (1 per cent). The test is not valid unless Calculate the content of C16H17N3O4S in the tablets.
the chromatogram obtained with reference solution (d) shows Storage. Store protected from light and moisture at a
three clearly separated spots. temperature not exceeding 30°.
Dissolution (2.5.2). Labelling. The label states the strength in terms of the
Apparatus. No 1 equivalent amount of anhydrous cephalexin.
271
CEPHALORIDINE IP 2007
Cephaloridine 1.25 ml of a solution prepared by decolorising a 0.5 per cent

v/v solution of bromine with potassium cyanide solution,
shaking and allowing to stand for 2 minutes, and sufficient
COO water to produce 25 ml and allow to stand for 25 minutes.
O
S O N N Measure the absorbance of the resulting solution at the
maximum at about 462 nm, using as the blank a solution
N S prepared in a similar manner but omitting the substance under
H H H examination (2.4.7). The absorbance is not more than that of a
solution prepared by treating 2.5 ml of a 0.005 per cent w/v
C19H17N3O4S2 Mol. Wt. 415.5 solution of pyridine in a similar manner.
Cephaloridine is (7R)-3-(1-pyridiniomethyl)-7-[(2-thienyl)- Sulphated ash (2.3.18). Not more than 0.2 per cent.
acetamido]-3-cephem-4-carboxylate (α-form or δ -form).
Water (2.3.43). Not more than 0.5 per cent w/w (α-form) and
Cephaloridine contains not less than 96.0 per cent and not not more than 3.0 per cent w/w (δ-form), determined on 0.25 g.
more than 102.0 per cent of C19H17N3O4S2, calculated on the Use as the solvent a mixture of equal volumes of dehydrated
anhydrous basis. methanol and dehydrated pyridine in place of methanol.
Description. A white or almost white, crystalline powder; odour, Assay. Weigh accurately about 60 mg and dissolve in sufficient
slight and resembling that of pyridine. water to produce 50.0 ml. Transfer 10.0 ml to a stoppered
flask, add 5 ml of 1 M sodium hydroxide and allow to stand for
Identification 20 minutes. Add 20 ml of a buffer solution containing 35.0 per
A. Determine by infrared absorption spectrophotometry (2.4.6). cent w/v of sodium acetate and 42.4 per cent v/v of glacial
Compare the spectrum with that obtained with cephaloridine acetic acid, 5 ml of 1 M hydrochloric acid and 25.0 ml of
(α-form) RS or cephaloridine (δ-form) RS or with the reference 0.01 M iodine, close the flask with a wet stopper and allow to
spectrum of cephaloridine (α-form) or cephaloridine (δ-form). stand for 3 hours in a water-bath at 30°, protected from light.
Titrate the excess of iodine with 0.02 M sodium thiosulphate
B. Mix 20 mg with a few drops of an 80 per cent v/v solution of using starch solution, added towards the end of the titration,
sulphuric acid containing 1 per cent v/v of nitric acid; a as indicator. To a further 10.0 ml of the solution add 20 ml of
bluish-green colour is produced. the buffer solution and 25.0 ml of 0.01 M iodine, allow to
C. To a 0.5 per cent w/v solution add 1 ml of chloramine stand for 3 hours in a water-bath at 30°, protected from light.
solution and 2 ml of 0.1 M sodium hydroxide; a dull red colour Titrate the excess of iodine with 0.02 M sodium thiosulphate
is produced which persists for 1 minute. using starch solution, added towards the end of the titration,
as indicator. The difference between the titrations represents
D. Gives the reactions of penicillins and cephalosporins (2.3.1).
the volume of 0.01 M iodine equivalent to the cephaloridine
Tests present. Calculate the content of C19H17N3O4S2 from the
difference obtained by simultaneously carrying out the Assay
Appearance of solution (2.4.1). Solution A is clear. using cephaloridine (δ-form) RS instead of the substance
pH (2.4.24). 4.0 to 6.0, determined in a 10.0 per cent w/v solution under examination.
(solution A) prepared by dissolving in carbon dioxide-free Cephaloridine intended for use in the manufacture of parenteral
water, warming to 30° and cooling to 20°. preparations complies with the following additional tests.
Specific optical rotation (2.4.22). +46.0° to +50.0°, determined Pyrogens. Complies with the test for pyrogens (2.2.8), using
at 25° in a 1.0 per cent w/v solution. not less than 50 mg per kg of the rabbit’s weight, dissolved in
1 ml of water for injection.
Light absorption. When examined in the range 230 nm to
360 nm (2.4.7) a 0.0012 per cent w/v solution shows absorption Sterility (2.2.11). Complies with the test for sterility.
maxima at about 240 nm and 255 nm; absorbance at the Storage. Store protected from light and moisture in a refrigerator
maximum at about 240 nm, 0.43 to 0.48. The ratio of the (8° to 15°). If the material is intended for use in the manufacture
absorbance at the maximum at about 240 nm to that at about of parenteral preparations, the container should be sterile,
255 nm is not more than 1.10. tamper-evident and sealed so as to exclude micro-organisms.
Pyridine. Dissolve about 25 mg in 10 ml of water and add Labelling. The label states (1) whether the contents are
2.5 ml of a buffer solution prepared by adjusting a 5 per cent Cephaloridine (α-form) or Cephaloridine (δ-form); (2) whether
w/v solution of disodium hydrogen phosphate to pH 6.0 with or not it is intended for use in the manufacture of injectable
phosphoric acid and adding 1 per cent v/v of aniline. Add preparations.
272
IP 2007 CEPHALORIDINE INJECTION
Cephaloridine Injection absorbance at the maximum at about 240 nm to that at about

255 nm is not more than 1.10.
Cephaloridine Injection is a sterile material consisting of
Cephaloridine with or without auxiliary substances. It is filled Pyridine. Dissolve about 25 mg in 10 ml of water and add
in a sealed container. 2.5 ml of a buffer solution prepared by adjusting a 5 per cent
w/v solution of disodium hydrogen phosphate to pH 6.0 with
The injection is constituted by dissolving the contents of the phosphoric acid and adding 1 per cent v/v of aniline. Add
sealed container in the requisite amount of Water for Injections 1.25 ml of a solution prepared by decolorising a 0.5 per cent
immediately before use. v/v solution of bromine with potassium cyanide solution,
The constituted solution complies with the requirements for shaking and allowing to stand for 2 minutes, and sufficient
Clarity of solution and Particulate matter stated under water to produce 25 ml and allow to stand for 25 minutes.
Parenteral Preparations (Injections). Measure the absorbance of the resulting solution at the
maximum at about 462 nm, using as the blank a solution
Storage. The constituted solution should be used immediately prepared in a similar manner but omitting the substance under
after preparation but, in any case, within the period examination (2.4.7). The absorbance is not more than that of a
recommended by the manufacturer. solution prepared by treating 2.5 ml of a 0.005 per cent w/v
Cephaloridine Injection contains not less than 90.0 per cent solution of pyridine in a similar manner.
and not more than 105.0 per cent of the stated amount of Sulphated ash (2.3.18). Not more than 0.2 per cent.
anhydrous cephaloridine, C19H17N3O4S2.
Water (2.3.43). Not more than 0.5 per cent w/w (α-form) and
Description. A white or almost white, crystalline powder; not more than 3.0 per cent w/w (δ-form), determined on 0.25 g.
odour, slight and resembling that of pyridine. Use as the solvent a mixture of equal volumes of dehydrated
The contents of the sealed container comply with the methanol and dehydrated pyridine in place of methanol.
requirements stated under Parenteral Preparations Assay. Determine the weight of the contents of 10 containers
(Powders for Injection) and with the following requirements. Weigh accurately a quantity of the mixed contents of the
10 containers containing about 60 mg of cephaloridine and
Identification dissolve in sufficient water to produce 50.0 ml. Transfer
A. Determine by infrared absorption spectrophotometry (2.4.6). 10.0 ml to a stoppered flask, add 5 ml of 1 M sodium hydroxide
Compare the spectrum with that obtained with cephaloridine and allow to stand for 20 minutes. Add 20 ml of a buffer solution
(α-form) RS or cephaloridine (δ-form) RS or with the reference containing 35.0 per cent w/v of sodium acetate and 42.4 per
spectrum of cephaloridine (α-form) or cephaloridine (δ-form). cent v/v of glacial acetic acid, 5 ml of 1 M hydrochloric acid
and 25.0 ml of 0.01 M iodine, close the flask with a wet stopper
B. Mix 20 mg with a few drops of an 80 per cent v/v solution of and allow to stand for 3 hours in a water-bath at 30°, protected
sulphuric acid containing 1 per cent v/v of nitric acid; a from light. Titrate the excess of iodine with 0.02 M sodium
bluish-green colour is produced. thiosulphate using starch solution, added towards the end
C. To a 0.5 per cent w/v solution add 1 ml of chloramine of the titration, as indicator. To a further 10.0 ml of the solution
solution and 2 ml of 0.1 M sodium hydroxide; a dull red colour add 20 ml of the buffer solution and 25.0 ml of 0.01 M iodine,
is produced which persists for 1 minute. allow to stand for 3 hours in a water-bath at 30°, protected
from light. Titrate the excess of iodine with 0.02 M sodium
D. Gives the reactions of penicillins and cephalosporins (2.3.1).
thiosulphate using starch solution, added towards the end of
the titration, as indicator. The difference between the titrations
Tests
represents the volume of 0.01 M iodine equivalent to the
Appearance of solution (2.4.1). Solution A is clear. cephaloridine present. Calculate the content of C19H17N3O4S2
from the difference obtained by simultaneously carrying out
the Assay using cephaloridine (δ−form) RS instead of the
(solution A) prepared by dissolving in carbon dioxide-free
substance under examination.
water, warming to 30° and cooling to 20°.
Cephaloridine intended for use in the manufacture of parenteral
preparations complies with the following additional tests.
Pyrogens. Complies with the test for pyrogens (2.2.8), using
Light absorption. When examined in the range 230 nm to
not less than 50 mg per kg of the rabbit’s weight, dissolved in
360 nm (2.4.7) a 0.0012 per cent w/v solution shows absorption
1 ml of water for injection.
maxima at about 240 nm and 255 nm; absorbance at the
maximum at about 240 nm, 0.43 to 0.48. The ratio of the Sterility (2.2.11). Complies with the test for sterility.
273
CETIRIZINE HYDROCHLORIDE IP 2007
Storage. Store protected from light and moisture at a Reference solution (b). Dissolve 10 mg of chlorphenamine
temperature not exceeding 30°. The constituted solution maleate RS in water and dilute to 5 ml with the same solvent.
should be used within 24 hours when stored at a temperature To 1 ml of the solution add 1 ml of reference solution (a).
not exceeding 20° or within 4 days when stored in a refrigerator Apply to the plate 5 µl of each solution. After development,
(2° to 8°). dry in a current of cold air and examine in ultraviolet light at
Labelling. The label states (1) the weight of Cephaloridine 254 nm. The principal spot in the chromatogram obtained with
contained in the sealed container; (2) whether the contents the test solution corresponds to the principal spot in the
are Cephaloridine (α-form) or Cephaloridine (δ-form). chromatogram obtained with reference solution (a). The test
solution (b) shows 2 clearly separated spots.
Cetirizine Hydrochloride D. It gives reaction A of chlorides (2.3.1).
Tests
Cl O COOH
N Appearance of solution. A 5.0 per cent w/v solution in carbon
N , 2HCl dioxide-free water (solution A) is clear (2.4.1) and not more
intensely coloured than reference solution BY7 (2.4.1).
(2.4.14).
C21H25ClN2O3,2HCl Mol. Wt. 461.8
Cetirizine Hydrochloride is [2-[4-[(4-chlorophenyl)
examination in the mobile phase and dilute to 100 ml with the
phenylmethyl]-1-piperazinyl]ethoxy]acetic acid.
mobile phase.
Cetirizine Hydrochloride contains not less than 99.0 per cent
and not more than 100.5 per cent of C21H25ClN2O3,2HCl,
w/v each of cetirizine dihydrochloride RS and (RS)-1-[(4-
chlorophenyl)phenylmethyl]piperazine RS ( cetirizine
Description. A white or almost white powder. impurity A) in the mobile phase. Dilute 1 ml of the solution to
Identification
Reference solution (b). Dilute 2 ml of the test solution to 50 ml
A. Determine by infrared absorption spectrophotometry (2.4.6). with the mobile phase. Dilute 5 ml of the solution to 100 ml
Compare the spectrum with that obtained with cetirizine with the mobile phase.
hydrochloride RS or with the reference spectrum of cetirizine
hydrochloride. Chromatographic system
B. Dissolve 20.0 mg in 50 ml of a 1.03 per cent w/v solution of octadecysilane bonded to porous silica (5 µm),
hydrochloric acid and dilute to 100.0 ml with the same acid. – mobile phase: a mixture of 0.4 volume of dilute sulphuric
Dilute 10.0 ml of this solution to 100.0 ml with the acid. acid, 6.6 volumes of water and 93 volumes of acetonitril,
When examined in the range 210 nm to 350 nm (2.4.7), the – flow rate. 1 ml per minute,
resulting solution shows an absorption maximum at about 231 – spectrophotometer set at 230 nm.
nm. The specific absorbance at 231 nm, 359 to 381. – a 20 µl loop injector.
C. Determine by thin-layer chromatography (2.4.17), coating Inject reference solution (a). The test is not valid unless the
the plate with silica gel GF 254. resolution between the peaks due to cetirizine and impurity A
is not less than 3 and the tailing factors are not more than 2.0.
Mobile phase. A mixture of 1 volume of ammonia, 10 volumes
of methanol and 90 volumes of dichloromethane. Inject the test solution and reference solution (b). Run the
chromatogram for 3 times the retention time of cetirizine. In
Test solution. Dissolve 10 mg of the substance under the chromatogram obtained with the test solution, the area of
examination in water and dilute to 5 ml with the same solvent. any impurity peak is not more than 0.5 times the area of the
Reference solution (a). Dissolve 10 mg of cetirizine principal peak in the chromatogram obtained with reference
hydrochloride RS in water and dilute to 5 ml with the same solution (b) (0.1 per cent). The sum of the areas of all such
solvent. peaks is not more than 1.5 times the area of the principal peak
274
IP 2007 CETIRIZINE TABLETS
in the chromatogram obtained with reference solution (b) (0.3 Hydrochloride, add 50 ml of the mobile phase, mix and dilute
per cent). Ignore any peak with an area 0.1 times the area of to 100 ml with the mobile phase.
the principal peak in the chromatogram obtained with reference Reference solution (a). A solution containing 0.02 per cent
solution (b) (0.02 per cent). w/v each of cetirizine hydrochloride RS and (RS)-1-[(4-
Sulphated ash (2.3.18). Not more than 0.2 per cent. chlorophenyl)phenylmethyl]piperazine RS ( cetirizine
impurity A) in the mobile phase. Dilute 1 ml of the solution to
on 1.0 g by drying in an oven at 100º to 105º.
Reference solution (b). Dilute 1 ml of the test solution to 100
Assay. Weigh accurately about 0.1 g, dissolve in 70 ml of a
ml with the mobile phase.
mixture of 30 volumes of water and 70 volumes of acetone.
Titrate with 0.1 M sodium hydroxide to the second point of Chromatographic system
inflexion. Determine the end-point potentiometrically (2.4.25). – a stainless steel column 25 cm x 4.6 mm, packed with
Carry out a blank titration. octadecysilane bonded to porous silica (5 µm),
– mobile phase: a mixture of 0.4 volume of dilute sulphuric
acid, 6.6 volumes of water and 93 volumes of
C21H27Cl3N2O3.
acetonitrile,
Storage. Store protected from light. – flow rate. 1 ml per minute,
Cetirizine Tablets resolution between the peaks due to cetirizine and impurity A
Cetirizine Hydrochloride Tablets is not less than 3 and the tailing factors are not more than 2.0.
Cetirizine Tablets contain not less than 90.0 per cent and not Inject the test solution and reference solution (b). Run the
more than 110.0 per cent of the stated amount of cetirizine chromatogram 3 times the retention time of cetirizine. In the
hydrochloride, C21H25ClN2O3.2HCl. chromatogram obtained with the test solution the area of any
impurity peak is not more than 0.5 times the area of the principal
Identification peak in the chromatogram obtained with reference solution
(b) (0.5 per cent). The sum of the areas of all such peaks is not
In the Assay, the principal peak in the chromatogram obtained more than 2 times the area of the principal peak in the
with the test solution corresponds to the peak in the chromatogram obtained with reference solution (b) (1.0 per
chromatogram obtained with the reference solution. cent). Ignore any peak with an area 0.1 times the area of the
Tests
solution (b) (0.05 per cent).
Dissolution (2.5.2). Uniformity of content. Comply with the test stated under
Apparatus No. 1 Tablets.
Medium. 900 ml of 0.1 M hydrochloric acid. Determine by liquid chromatography (2.4.17), as described
Speed and time. 100 rpm and 45 minutes. under Assay, using the following solution as the test solution.
Withdraw a suitable volume of the medium and filter. Measure Test solution. Disperse 1 tablet in the mobile phase, mix and
the absorbance of the filtrate, suitably diluted with the dilute to 100.0 ml with the mobile phase, filter. Dilute 5.0 ml of
dissolution medium if necessary, at the maximum at about 230 the solution to 10.0 ml with mobile phase.
nm (2.4.7). Calculate the content of C21H25ClN2O3.2HCl in the Other tests. Comply with the tests stated under Tablets.
medium from the absorbance obtained from a solution of known
concentration of cetirizine hydrochloride RS in the same Assay. Determine by liquid chromatography (2.4.14).
medium. Test solution. Weigh and powder 20 tablets. Weigh accurately
a quantity of the powder containing about 25 mg of Cetirizine
Hydrochloride, add the mobile phase, mix and dilute to 50.0 ml
C21H25ClN2O3.2HCl.
with the mobile phase, filter. Dilute 1.0 ml of the solution to
Related substances. Determine by liquid chromatography 10.0 ml with mobile phase.
(2.4.14).
Reference solution. A 0.05 per cent w/v solution of cetirizine
Test solution. Weigh and powder 20 tablets. Weigh accurately hydrochloride RS in the mobile phase. Dilute 1.0 ml of the
a quantity of the powder containing 20 mg of Cetirizine solution to 10.0 ml with the mobile phase.
275
CETOSTEARYL ALCOHOL IP 2007
Chromatographic system Cetrimide

octadecylsilane chemically bonded to porous silica (5 Cetrimide consists chiefly of tetradecyltrimethylammonium
µm), bromide together with smaller amounts of dodecyl- and
– mobile phase: dissolve 0.19 g of heptane sulphonic acid hexadecyltrimethylammonium bromides.
sodium salt in 300 ml water add 700 ml acetonitrile and Cetrimide contains not less than 96.0 per cent and not more
mix. Adjust pH to 3.2 with 0.05 M sulphuric acid, filter, than 101.0 per cent of alkyltrimethylammonium bromides,
– flow rate. 1.2 ml per minute, calculated as C17H38BrN (336.4) on the dried basis.
– a 20 µl loop injector. Description. A white or creamy-white, voluminous, free-
flowing powder; odour, faint and characteristic.
tailing factor is not more than 2.0, the column efficiency in not Identification
less than 2000 theoretical plates and the relative standard
deviation for replicate injections is not more than 2.0 per cent. A. To 10 ml of a 1 per cent w/v solution add 2 ml of potassium
ferricyanide solution; a yellow precipitate is produced.
B. To 10 ml of a 1 per cent w/v solution add 2 ml of a 10 per cent
Calculate the content of C21H25ClN2O3.2HCl in the tablets. w/v solution of sodium silicate; a white flocculent precipitate
Storage. Store protected from moisture, at a temperature not is produced.
exceeding 30º. C. To 10 ml of a 1 per cent w/v solution add 2 ml of dilute nitric
acid; a yellow precipitate is produced. Filter and to the filtrate
add 2 ml of dilute nitric acid and 1 ml of silver nitrate solution;
a yellow precipitate is produced.
Cetostearyl Alcohol D. Dissolve 0.25 g in sufficient ethanol (95 per cent) to
Cetostearyl Alcohol is a mixture of solid aliphatic alcohols produce 25 ml. Absorbance of the resulting solution between
consisting chiefly of stearyl and cetyl alcohols. 260 nm and 280 nm is not more than 0.05 (2.4.7).
Description. A white or pale yellow, wax like mass, plates,
Tests
flakes or granules.
Tests (2.4.1) and colourless (2.4.1).
Melting range (2.4.21). 47° to 56°, determined by Method II. Acidity or alkalinity. Dissolve 1.0 g in 50 ml of water and add
Introduce the substance under examination into the capillary 2 drops of bromocresol purple solution. Not more than 0.1 ml
tubes and allow to stand at 2° to 8° for 12 hours before carrying of either 0.1 M hydrochloric acid or 0.1 M sodium hydroxide
out the determination. is required to change the colour of the solution.
Appearance of solution. Dissolve 0.5 g in 20 ml of boiling Amine salts. Carry out the Assay described below using a
ethanol (95 per cent). The solution is clear (2.4.1) and not further 25.0 ml of the original solution and 10 ml of 0.1 M
more intensely coloured than reference solution BS6 (2.4.1) hydrochloric acid instead of the 0.1 M sodium hydroxide.
The difference between the volume of 0.05 M potassium
Acid value (2.3.23). Not more than 1.0.
iodate required in the titration and that required in the Assay
Hydroxyl value (2.3.27). 208 to 228. is not more than 1.0 ml for each g of the substance used.
Saponification value (2.3.37). Not more than 2.0. Sulphated ash (2.3.18). Not more than 0.5 per cent.
Iodine value (2.3.28). Not more than 3.0, determined by Method Loss on drying (2.4.19). Not more than 2.0 per cent, determined
B in a 8.0 per cent w/v solution in chloroform. on 1.0 g by drying in an oven at 105° for 2 hours.
Hydrocarbons. Dissolve 2.0 g in 100 ml of light petroleum Assay. Weigh accurately about 2.0 g and dissolve in sufficient
(40° to 60°), warming slightly if necessary, and transfer the water to produce 100.0 ml. Transfer 25.0 ml of the solution to
solution to a column (25 cm x 10 mm) of anhydrous alumina a separator, add 25 ml of chloroform, 10 ml of 0.1 M sodium
which has been slurried with light petroleum (40° to 60°). hydroxide and 10.0 ml of a freshly prepared 5.0 per cent w/v
Elute with two portions, each of 50 ml, of light petroleum solution of potassium iodide. Shake well, allow to separate,
(40° to 60°) into a flask, remove the light petroleum and dry at and discard the chloroform layer. Shake the aqueous solution
80°; the residue weighs not more than 30 mg. with three quantities, each of 10 ml, of chloroform and discard
276
IP 2007 ACTIVATED CHARCOAL
the chloroform solution. Add 40 ml of hydrochloric acid, allow B. Complies with the test for Adsorbing power.
to cool and titrate with 0.05 M potassium iodate until the
deep brown colour is almost discharged. Add 2 ml of chloroform Tests
and continue the titration, with shaking, until the chloroform Acidity or alkalinity. Boil 2.0 g with 40 ml of water for 5 minutes.
layer no longer changes colour. Carry out a blank titration on Cool, restore to the original volume with carbon dioxide-free
a mixture of 20 ml of water, 10.0 ml of the freshly prepared water and filter, discarding the first 20 ml of the filtrate. To
potassium iodide solution and 40 ml of hydrochloric acid. 10 ml of the filtrate add 0.25 ml of bromothymol blue solution
The difference between the titrations represents the amount and 0.25 ml of 0.02 M sodium hydroxide. The solution is blue
of potassium iodate required. and not more than 0.75 ml of 0.02 M hydrochloric acid is
1 ml of 0.05 M potassium iodate is equivalent to 0.03364 g of required to change the colour to yellow.
C17H38BrN. Acid-soluble substances. Boil 1.0 g with a mixture of 20 ml of
water and 5 ml of hydrochloric acid for 5 minutes, filter whilst
hot and collect the filtrate in a previously weighed porcelain
Cetyl Alcohol crucible, wash the residue with 10 ml of hot water, adding the
washing to the filtrate. To the combined filtrate and washing
Palmityl Alcohol; n-Hexadecyl Alcohol; 1-Hexadecanol add 1 ml of hydrochloric acid, evaporate to dryness and ignite
Cetyl Alcohol is a mixture of solid alcohols consisting mainly gently to constant weight; the residue weighs not more than
of 1-hexadecanol, C16H34O. 30 mg.
Description. A white, unctuous mass, powder, flakes or Ethanol-soluble substances. Boil 2.0 g with 50 ml of ethanol
granules; odour, slight. (95 per cent) under a reflux condenser for 10 minutes. Filter
immediately, cool and adjust the volume to 50 ml with ethanol
Tests (95 per cent). The filtrate is not more intensely coloured than
reference solution BYS6 or YS6 (2.4.1). Evaporate 40 ml of the
Melting range (2.4.21). 46° to 52°, determined by Method II. filtrate to dryness; the residue, after drying to constant weight
Introduce the substance under examination into the capillary at 105°, weighs not more than 8 mg.
tubes and allow to stand at 2° to 8° for 12 hours before carrying
Alkali-soluble coloured matter. Boil 0.25 g with 10 ml of 2 M
out the determination.
sodium hydroxide for 1 minute, cool and filter; the filtrate,
Appearance of solution. Dissolve 0.5 g in boiling ethanol when diluted to 10 ml with water, is not more intensely coloured
(95 per cent), cool and dilute to 20 ml with the same solvent. than reference solution GYS4, (2.4.1).
The resulting solution is clear (2.4.1) and not more intensely
Chlorides (2.3.12). Boil 3.0 g with 75 ml of water for 5 minutes,
cool. Dilute to 100.0 ml with water and filter; 6.0 ml of the
Acid value (2.3.23). Not more than 1.0. filtrate complies with the limit test for chlorides. (0.14 per cent).
Hydroxyl value (2.3.27). 218 to 238. Sulphates (2.3.17). 10.0 ml of the filtrate obtained in the test
for Chloride complies with the limit test for sulphates
Saponification value (2.3.37). Not more than 2.0.
(450 ppm).
Iodine value (2.3.28). Not more than 2.0, determined by Method
Sulphide. Heat 1.0 g with a mixture of 20 ml of water and 5 ml
B in a 8.0 per cent w/v solution in chloroform.
of 7 M hydrochloric acid to boiling; the fumes evolved do
not turn lead acetate paper brown.
Uncarbonised constituents. Boil 0.25 g with 10 ml of 1 M sodium
Activated Charcoal hydroxide for few seconds and filter; the filtrate is colourless.
Decolorising Charcoal Copper. Determine by atomic absorption spectrophotometry
Activated Charcoal is obtained from vegetable matter by (2.4.2), measuring at 325.0 nm using an air-acetylene flame and
suitable carbonisation processes intended to confer a high a solution prepared in the following manner. Boil 2.0 g with
adsorbing power. 50 ml of 2 M hydrochloric acid under a reflux condenser for
1 hour. Filter, wash the filter with 2 M hydrochloric acid and
Description. A light, black powder, free from grittiness; evaporate the combined filtrate to dryness on a water-bath.
odourless. Dissolve the residue in sufficient 0.1 M hydrochloric acid to
Identification produce 50.0 ml. Use copper solution AAS, suitably diluted
with 0.1 M hydrochloric acid, for preparing the standard
A. When heated to redness, burns slowly without flame. solutions (25 ppm).
277
CHLORAMBUCIL IP 2007
Reserve the solution for the tests for Lead and Zinc. CAUTION — Chlorambucil must be handled with care;
Lead. Determine by atomic absorption spectrophotometry contact with the skin and inhalation of airborne particles
(2.4.2), measuring at 283.3 nm or 217.0 nm using an air-acetylene must be avoided.
flame. Use the solution prepared in the test for Copper as the Identification
test solution and lead solution AAS, suitably diluted with
0.1 M hydrochloric acid, for preparing the standard solutions Test A may be omitted if tests B and C are carried out. Tests B
(10 ppm). and C may be omitted if test A is carried out.
Zinc. Determine by atomic absorption spectrophotometry A. Determine by infrared absorption spectrophotometry (2.4.6).
(2.4.2), measuring at 214.0 nm using an air-acetylene flame. Compare the spectrum with that obtained with chlorambucil
Use the solution prepared in the test for Copper as the test RS.
solution and zinc solution AAS, suitably diluted with 0.1 M B. Shake 0.4 g with 10 ml of 2 M hydrochloric acid and allow
hydrochloric acid, for preparing the standard solutions to stand for 30 minutes, shaking occasionally. Filter, wash the
(25 ppm). residue with two quantities, each of 10 ml, of water and add
Sulphated ash (2.3.18). Not more than 5.0 per cent. 0.5 ml of potassium mercuri-iodide solution to 10 ml of the
mixed filtrate and washings; a buff precipitate is produced. To
a further 10 ml add 0.5 ml of potassium permanganate
solution; the purple colour is immediately discharged.
Adsorbing power. Not less than 40 per cent of its own weight C. Dissolve 50 mg in 5 ml of acetone and dilute to 10 ml with
of phenazone, calculated on the dried basis, determined by water. Add 0.05 ml of 2 M nitric acid and 0.2 ml of dilute silver
the following method. To 0.3 g add 25 ml of a freshly prepared nitrate solution; no opalescence is produced immediately.
1 per cent w/v solution of phenazone, shake thoroughly for Heat on a water-bath; an opalescence is produced.
15 minutes, filter and discard the first 5 ml of the filtrate. To
10 ml of the filtrate add 1 g of potassium bromide and 20 ml of Tests
2 M hydrochloric acid and titrate with 0.0167 M potassium
bromate, using 0.1 ml of methyl red solution as indicator, until Related substances. Carry out all operations as rapidly as
the colour changes from reddish pink to yellowish pink and possible, protected from light, and prepare the solutions
titrate slowly towards the end of the titration (a ml). Repeat immediately before use.
the titration using 10 ml of the phenazone solution beginning Determine by thin-layer chromatography (2.4.17), coating the
at the words “add 1 g..... titration” (b ml). Calculate the plate with silica gel GF254.
percentage of phenazone adsorbed with reference to the dried Mobile phase. A mixture of 40 volumes of toluene, 25 volumes
substance using the expression 2.353 (a-b)/w where w is the of methanol, 20 volumes of 2-butanone and 20 volumes of
weight, in g, of the substance under examination. n-heptane.
Storage. Store protected from moisture. Test solution. A 2 per cent w/v solution in acetone.
with acetone.
Chlorambucil Reference solution (b). Dilute 5 ml of reference solution (a) to
20 ml with acetone.
COOH Apply to the plate 5 µl of each solution. After development,
Cl dry the plate in air and examine in ultraviolet light at 254 nm.
N
Cl test solution is not more intense than the spot in the
C14H19Cl2NO2 Mol. Wt. 304.2 more than one such spot is more intense than the spot in the
Chlorambucil is 4-[4-bis(2-chloroethyl)amino]phenylbutyric chromatogram obtained with reference solution (b).
acid. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Chlorambucil contains not less than 98.0 per cent and not Water (2.3.43). Not more than 0.5 per cent, determined on 1.0 g
more than 101.0 per cent of C14H19Cl2NO2, calculated on the
anhydrous basis.
acetone, add 10 ml of water and titrate with 0.1 M sodium
Description. A white, crystalline powder. hydroxide using dilute phenolphthalein solution as indicator.
278
IP 2007 CHLORAMPHENICOL
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03042 g of about 10 mg of Chlorambucil in a mixture of 25 ml of 0.1 M

C14H19Cl2NO2. hydrochloric acid and 100 ml of acetonitrile by mixing in an
Storage. Store protected from light. ultrasonic bath for at least 10 minutes. Dilute to 250.0 ml with
acetonitrile and filter the solution, preferably through a glass
microfibre filter paper (such as Whatman GF/C), discarding
the first 20 ml of the filtrate. Dilute 50.0 ml of the filtrate to
Chlorambucil Tablets 100.0 ml with a mixture of 90 volumes of acetonitrile and
Chlorambucil Tablets contain not less than 90.0 per cent and 10 volumes of 0.1 M hydrochloric acid.
not more than 110.0 per cent of the stated amount of Reference solution. A 0.002 per cent w/v solution of
chlorambucil, C14H19Cl2NO2. The tablets are coated. chlorambucil RS in a mixture of 90 volumes of acetonitrile
and 10 volumes of 0.1 M hydrochloric acid.
Identification
Carry out the chromatographic procedure described under
Shake 0.4 g of the powdered tablets with 10 ml of 2 M Uniformity of content.
hydrochloric acid and allow to stand for 30 minutes, shaking
Calculate the content of C14H19Cl2NO2 in the tablets.
occasionally. Filter, wash the residue with two quantities, each
of 10 ml, of water and add 0.5 ml of potassium mercuri-iodide
solution to 10 ml of the mixed filtrate and washings; a buff
precipitate is produced. To a further 10 ml add 0.5 ml of Chloramphenicol
potassium permanganate solution; the purple colour is
immediately discharged. OH Cl
H
Tests N
Cl
O
Uniformity of content. Comply with the test stated under O 2N OH
Tablets.
Determine by liquid chromatography (2.4.14). C11H12Cl2N2O5 Mol. Wt. 323.1
Test solution. Dissolve one tablet as completely as possible Chloramphenicol is 2,2-dichloro-N-[(1R,2R)-2-hydroxy-1-
in 10 ml of 0.1 M hydrochloric acid, add 40 ml of acetonitrile hydroxymethyl-2-(4-nitrophenyl)ethyl]acetamide. It is
and mix in an ultrasonic bath for 5 minutes. Add sufficient produced by the growth of certain strains of Streptomyces
acetonitrile to produce a solution containing 0.002 per cent venezuelae in a suitable medium, but is normally prepared
w/v of Chlorambucil. Filter the solution, preferably through a by synthesis.
glass microfibre filter paper (such as Whatman GF/C), Chloramphenicol contains not less than 98.0 per cent and not
discarding the first 20 ml of the filtrate, and use the filtrate. more than 102.0 per cent of C11H12Cl2N2O5, calculated on the
Reference solution. A 0.002 per cent w/v solution of dried basis.
chlorambucil RS in a mixture of 90 volumes of acetonitrile Description. A white to greyish-white or yellowish-white, fine
and 10 volumes of 0.1 M hydrochloric acid. crystalline powder or fine-crystals, needles or elongated plates;
Chromatographic system odourless.
Identification
– mobile phase: a mixture of 60 volumes of acetonitrile Test A may be omitted if tests B, C and D are carried out. Tests
and 40 volumes of 0.02 M potassium dihydrogen B, C and D may be omitted if test A is carried out.
phosphate,
Compare the spectrum with that obtained with
chloramphenicol RS or with the reference spectrum of
chloramphenicol.
Calculate the content of C14H19Cl2NO2 in the tablet.
Other tests. Comply with the tests stated under Tablets. chromatogram obtained with the test solution corresponds to
Assay. Determine by liquid chromatography (2.4.14). that in the chromatogram obtained with reference solution (a).
Test solution. Weigh and powder 20 tablets. Dissolve as C. Dissolve 10 mg in 1 ml of ethanol (50 per cent), add 3 ml of
completely as possible a quantity of the powder containing a 1 per cent w/v solution of calcium chloride and 50 mg of
279
CHLORAMPHENICOL CAPSULES IP 2007
zinc powder and heat on a water-bath for 10 minutes. Decant Calculate the content of C11H12Cl2N2O5 taking 297 as the
the clear supernatant liquid into a test-tube, add 0.1 g of specific absorbance at 278 nm.
anhydrous sodium acetate and 0.1 ml of benzoyl chloride, Chloramphenicol intended for use in the manufacture of
shake for 1 minute and add 0.5 ml of a 10.5 per cent w/v solution parenteral preparations without a further process for the
of ferric chloride hexahydrate and, if necessary, add sufficient removal of bacterial endotoxins complies with the following
dilute hydrochloric acid to produce a clear solution; a red- additional requirement.
violet to purple colour is produced. Repeat the test omitting
the zinc powder; no red colour is produced. Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit
per mg of chloramphenicol.
D. Heat 50 mg with 2 ml of ethanolic potassium hydroxide
solution in a covered test-tube on a water-bath for 15 minutes; Chloramphenicol intended for use in the manufacture of
the resulting solution gives the reactions of chlorides (2.3.1). parenteral or ophthalmic preparations without a further
Tests additional requirement.
pH (2.4.24). 4.5 to 7.5, determined in a suspension prepared by
shaking 50 mg with 10 ml of carbon dioxide-free water. Storage. Store protected from light and moisture. If the material
is intended for use in the manufacture of parenteral or
ophthalmic preparations without a further appropriate
in a 5.0 per cent w/v solution in ethanol.
procedure of sterilisation, the container should be sterile,
Related substances. Determine by thin-layer chromatography tamper-evident and sealed so as to exclude micro-organisms.
(2.4.17), coating the plate with silica gel GF254. Labelling. The label states whether or not the contents are
Mobile phase. A mixture of 90 volumes of chloroform, intended for use in the manufacture of parenteral or ophthalmic
10 volumes of methanol and 1 volume of water. preparations.
Test solution. Dissolve 1 g of the substance under examination
in 100 ml of acetone.
Chloramphenicol Capsules
Reference solution (a). A 1 per cent w/v solution of
chloramphenicol RS in acetone. Chloramphenicol Capsules contain not less than 92.5 per cent
Reference solution (b). Dilute 0.5 ml of reference solution (a) chloramphenicol, C11H12Cl2N2O5.
to 100 ml with acetone.
Apply to the plate 1 µl and 20 µl of the test solution, 1 µl of
Identification
reference solution (a) and 20 µl of reference solution (b). After Suspend a quantity of the contents of the capsules containing
development, dry the plate in air and examine in ultraviolet about 1.25 g of Chloramphenicol in 60 ml of water and extract
light at 254 nm. Any secondary spot in the chromatogram with two quantities, each of 20 ml, of light petroleum (60° to
obtained with 20 µl of the test solution is not more intense 80°) or light petroleum (100° to 120°). Wash the combined
than the spot in the chromatogram obtained with reference extracts with two quantities, each of 15 ml, of water, add the
solution (b). washings to the aqueous layer, extract with four quantities,
Chlorides (2.3.12). To 2.0 g add 20 ml of water and 10 ml of each of 50 ml, of ether and remove the ether from the combined
nitric acid and shake for 5 minutes. Filter through a filter extracts. The residue, after drying to constant weight at 105°,
paper previously washed by filtering 5-ml quantities of water complies with the following tests.
until 5 ml of the filtrate is no longer opalescent on addition of A. Determine by infrared absorption spectrophotometry (2.4.6).
0.1 ml of nitric acid and 0.1 ml of a 4.25 per cent w/v solution Compare the spectrum with that obtained with
of silver nitrate. The resulting filtrate complies with the limit chloramphenicol RS or with the reference spectrum of
test for chlorides (125 ppm). chloramphenicol.
Sulphated ash (2.3.18). Not more than 0.1 per cent. B. Heat 50 mg with 2 ml of ethanolic potassium hydroxide
Loss on drying (2.4.19). Not more than 0.5 per cent, determined solution in a covered test-tube on a water-bath for 15 minutes;
on 1.0 g by drying in an oven at 105°. the resulting solution gives the reactions of chlorides (2.3.1).
Assay. Weigh accurately about 0.125 g and dissolve in Tests

sufficient water to produce 250.0 ml. Dilute 10.0 ml with Specific optical rotation (2.4.22). +17.0° to +20.0°, determined
sufficient water to produce 250.0 ml. Measure the absorbance in a 5.0 per cent w/v solution in ethanol of the residue obtained
of the resulting solution at the maximum at about 278 nm (2.4.7). in the test for Identification.
280
IP 2007 CHLORAMPHENICOL EYE OINTMENT
Dissolution (2.5.2). Apply to the plate 1µl of each solution. Allow the mobile
Apparatus. No. 1 phase to rise 10 cm. Dry the plate in a current of warm air and
Withdraw a suitable volume of the medium and filter promptly B. Dissolve 10 mg in 2 ml of ethanol (50 per cent), add 4.5 ml
through a membrane filter disc having an average pore diameter of 1 M sulphuric acid and 50 mg of zinc powder and allow to
not greater than 1.0 µm, rejecting the first 1 ml of the filtrate. stand for 10 minutes. Decant the supernatant liquid or filter, if
Dilute 5.0 ml of the filtrate to 100.0 ml with the same solvent. necessary. Cool the resulting solution in ice and add 0.5 ml of
Measure the absorbance of the resulting solution at the sodium nitrite solution and, after 2 minutes, 1 g of urea
maximum at about 278 nm (2.4.7). Calculate the content of followed by 1 ml of 2-naphthol solution and 2 ml of 10 M
C11H12Cl2N2O5 taking 297 as the specific absorbance at sodium hydroxide; a red colour is produced. Repeat the test
278 nm. omitting the zinc powder; no red colour is produced.
C11H12Cl2N2O5. Tests
Other tests. Comply with the tests stated under Capsules. pH (2.4.24). 7.0 to 7.5.
Assay. Weigh accurately a quantity of the mixed contents of Assay. Determine by liquid chromatography (2.4.14).
20 capsules containing about 0.2 g of Chloramphenicol, Test solution. Dilute a suitable volume of the eye drops
dissolve in 800 ml of water, warming if necessary to effect containing about 50 mg of chloramphenicol to 100.0 ml with
solution and add sufficient water to produce 1000.0 ml. Dilute the mobile phase. Dilute 5.0 ml of this solution to 25.0 ml with
10.0 ml of this solution to 100.0 ml with water and measure the the mobile phase and filter through a 0.5 mm or finer porosity
absorbance of the resulting solution at the maximum at about filter and use the clear filtrate.
278 nm (2.4.7). Calculate the content of C11H12Cl2N2O5, taking
297 as the specific absorbance at 278 nm.
chloramphenicol RS in the mobile phase. Filter this solution
Storage. Store protected from moisture. through a 0.5 mm or finer porousity filter and use the clear
filtrate.
Chloramphenicol Eye Drops octadecylsilyl silica gel (5 µm),
Chloramphenicol Eye Drops are a sterile solution of – mobile phase: a mixture of 55 volumes of water,
Chloramphenicol in Purified water. 45 volumes of methanol and 0.1 volume of glacial acetic
acid,
Chloramphenicol Eye Drops contain not less than 90.0 per – flow rate. 1 ml per minute,
cent and not more than 130.0 per cent of the stated amount of – spectrophotometer set at 280 nm,
chloramphenicol, C11H12Cl2N2O5. – a 20 µl loop injector.
Identification Inject alternately the test solution and the reference solution.
Calculate the content of C11H12Cl2N2O5 in the drops.
To a volume containing 50 mg of Chloramphenicol add 15 ml
of water and extract with four quantities, each of 25 ml, of Storage. Store in light resistant containers at a temperature
ether. Combine the extracts and evaporate to dryness. The not exceeding 30°.
residue complies with the following tests.
the plate with silica gel G F254. Chloramphenicol Eye Ointment
Mobile phase. A mixture of 90 volumes of chloroform, Chloramphenicol Eye Ointment contains not less than
10 volumes of methanol and 1 volume of water. 90.0 per cent and not more than 120.0 per cent of the stated
amount of chloramphenicol, C11H12Cl2N2O5.
Test solution. Dissolve 0.1 g of the residue in sufficient ethanol
(95 per cent) to produce 10 ml. Identification
Reference solution. Dissolve 0.1 g of chloramphenicol RS in Mix a quantity of the ointment containing 30 mg of
sufficient ethanol (95 per cent) to produce 10 ml. Chloramphenicol with 10 ml of light petroleum (40° to 60°),
281
CHLORAMPHENICOL PALMITATE IP 2007
centrifuge and discard the supernatant liquid. Repeat this Chloramphenicol Palmitate
procedure using three quantities, each of 10 ml, of the same
solvent. Combine the extracts and evaporate to dryness. The OH Cl
residue complies with the following tests. H
N
A. Determine by infrared absorption spectrophotometry (2.4.6). Cl
O
Compare the spectrum with that obtained with O 2N O
chloramphenicol RS or with the reference spectrum of
chloramphenicol. O (CH2)14CH3
B. Heat 50 mg with 2 ml of ethanolic potassium hydroxide C27H42Cl2N2O6 Mol. Wt. 561.5
solution in a covered test-tube on a water-bath for 15 minutes;
the resulting solution gives the reactions of chlorides (2.3.1). Chloramphenicol Palmitate is (2R,3R)-2-(2,2-
dichloroacetamido)-3-hydroxy-3-(4-nitrophenyl)propyl
Tests hexadecanoate.
Other tests. Complies with the tests stated under Eye Chloramphenicol Palmitate contains not less than 97.0 per
Ointments. cent and not more than 103.0 per cent of C27H42Cl2N2O6,
Description. A fine, white or almost white, unctuous powder;
Test solution. Transfer an accurately weighed quantity of the odour, faint.
ointment, containing about 25 mg of Chloramphenicol, to a
suitable conical flask, add 20 ml of cyclohexane, mix with the Identification
aid of ultrasound for about 2 minutes add 60 ml of methanol, A. When examined in the range 230 nm to 360 nm (2.4.7), a
and mix. Filter this mixture, collecting the filtrate in a 100- ml 0.003 per cent w/v solution in ethanol (95 per cent) shows an
volumetric flask. Wash the filter with methanol, collecting the absorption maximum only at about 271 nm; absorbance at
washings in the volumetric flask. Dilute with methanol to about 271 nm, about 0.53.
volume, and mix. Transfer 50.0 ml of the resulting solution to a
suitable round-bottom flask, and evaporate to dryness by B. Determine by thin-layer chromatography (2.4.17), coating
rotating the flask under vacuum in a water-bath at 35°. Dissolve the plate with silanised silica gel H.
the residue in 50.0 ml of methanol. Transfer 10.0 ml of the Mobile phase. A mixture of 70 volumes of ethanol (95 per
resulting solution to a 25 ml-volumetric flask, dilute with the cent) and 30 volumes of a 10 per cent w/v solution of
mobile phase to volume, and mix. Filter a portion of this solution ammonium acetate.
through a 0.5 µm or finer porosity filter, and use the clear
filtrate.
examination in a mixture of 1 ml of 1 M sodium hydroxide and
Reference solution. A 0.01 per cent w/v solution of 5 ml of acetone, allow to stand for 30 minutes and add 1.1 ml of
chloramphenicol RS in the mobile phase. Filter this solution 1 M hydrochloric acid and 3 ml of acetone.
through a 0.5 µm or finer porosity filter and use the clear Reference solution (a). A 0.2 per cent w/v solution of
filtrate. chloramphenicol RS in acetone.
Chromatographic system Reference solution (b). A 0.2 per cent w/v solution of palmitic
– a stainless steel column 25 cm x 4.6 mm, packed with acid in acetone.
Reference solution (c). A 0.2 per cent w/v solution of the
substance under examination in acetone.
45 volumes of methanol and 0.1 volume of glacial acetic
acid, Apply to the plate 4 µl of each solution. After development,
– flow rate. 1 ml per minute, dry the plate in air and spray with a solution containing
– spectrophotometer set at 280 nm, 0.02 per cent w/v of 2,7-dichlorofluorescein and 0.01 per cent
w/v of rhodamine B in ethanol (95 per cent). Allow the plate
to dry in air and examine in ultraviolet light at 254 nm. The
Inject alternately the test solution and the reference solution. chromatogram obtained with the test solution shows three
Calculate the content of C11H12Cl2N2O5 in the ointment. spots corresponding in position to the principal spots in
chromatograms obtained with reference solutions (a), (b) and
Storage. Store at a temperature not exceeding 30°. (c).
282
IP 2007 CHLORAMPHENICOL ORAL SUSPENSION
C. Dissolve 10 mg in 4 ml of ethanol (95 per cent) add 1 ml of Loss on drying (2.4.19). Not more than 0.5 per cent, determined
1 M sulphuric acid and 50 mg of zinc powder and allow to on 1.0 g by drying in an oven over phosphorus pentoxide at
stand for 10 minutes. Filter, cool the filtrate in ice and add 80° at a pressure not exceeding 0.1 kPa for 3 hours.
0.5 ml of sodium nitrite solution and, after 2 minutes, 1 g of Assay. Weigh accurately about 60 mg and dissolve in sufficient
urea followed by 1 ml of 2-naphthol solution and 2 ml of 10 M ethanol (95 per cent) to produce 100.0 ml. Dilute 10.0 ml of
sodium hydroxide; a red colour develops. Repeat the test this solution to 200.0 ml with ethanol (95 per cent) and measure
omitting the zinc powder; no red colour is produced. the absorbance of the resulting solution at the maximum at
D. Heat 50 mg with 2 ml of ethanolic potassium hydroxide about 271 nm (2.4.7). Calculate the content of C27H42Cl2N2O6
solution in a covered test-tube on a water-bath for 15 minutes; taking 178 as the specific absorbance at 271 nm.
the resulting solution gives the reactions of chlorides (2.3.1).
Tests
Free acid. Dissolve 1.0 g by warming to 35° in 5 ml of a mixture
of a equal volumes of ethanol (95 per cent) and ether and Chloramphenicol Oral Suspension
add 0.2 ml of phenolphthalein solution; not more than 0.4 ml
Chloramphenicol Palmitate Oral Suspension;
of 0.1 M sodium hydroxide is required to produce a pink
colour persisting for 30 seconds.
Chloramphenicol Palmitate Mixture
Chloramphenicol Oral Suspension is a suspension of
Chloramphenicol Palmitate in a suitable flavoured vehicle.
Related substances. Determine by thin-layer chromatography Chloramphenicol Oral Suspension contains not less than
(2.4.17), coating the plate with silica gel GF254. 95.0 per cent and not more than 115.0 per cent of the stated
amount of chloramphenicol, C11H12Cl2N2O5.
40 volumes of chloroform and 10 volumes of methanol. Identification
Test solution. Dissolve 1 g of the substance under examination Extract a quantity of the suspension containing about 7.5 mg
in 100 ml of acetone. of chloramphenicol with 10 ml of chloroform and carefully
Reference solution. Dilute 2 ml of the test solution to 100 ml evaporate the clear chloroform solution on a water-bath to
with acetone. dryness. Dissolve the residue in 250 ml of ethanol (95 per cent.
When examined in the range 230 nm to 360 nm (2.4.7) the resulting
Apply to the plate 10 µl of each solution. After development, solution shows an absorption maximum only at about 271 nm.
Any secondary spot in the chromatogram obtained with the Tests
test solution is not more intense than the spot in the
pH (2.4.24). 4.5 to 7.0.
Polymorph A. To a volume of the suspension containing
Free chloramphenicol. Not more than 450 ppm, determined
125 mg of chloramphenicol add 35 ml of water, mix, centrifuge
by the following method. Dissolve, with the aid of gentle heat,
for 40 minutes at not less than 18,000 rpm and discard the
1.0 g in 80 ml of xylene, cool and extract with three successive
supernatant liquid. Wash the residue by adding 2 ml of water,
quantities, each of 15 ml, of water; discard the xylene and
triturating to form a paste, adding 18 ml of water, mixing
dilute the combined aqueous extracts to 50 ml with water.
thoroughly centrifuging and discarding the supernatant liquid.
Extract the solution with 10 ml of carbon tetrachloride, allow
Wash the residue twice more in a similar manner, dry at 20° for
to separate, discard the carbon tetrachloride and centrifuge a
16 hours at a pressure not exceeding 0.7 kPa and grind to a
portion of the aqueous solution. Measure the absorbance of
fine powder. Prepare a mull of the residue by triturating a small
the clear aqueous solution at the maximum at about 278 nm,
quantity with about twice its weight of liquid paraffin until a
using as the blank a solution obtained by repeating the
smooth creamy paste is obtained. Determine by infrared
procedure without the substance under examination; the
absorption spectrophotometry (2.4.6) over the range 770 cm-1
absorbance of this blank solution must not be greater than
to 910 cm–1 using conditions such that between 20 per cent
0.05 (2.4.7). Calculate the content of free chloramphenicol, in
and 30 per cent transmittance occurs at 810 cm–1 to 910 cm–1.
ppm, from the expression (A x 104)/5.96, where A is the
Repeat the operation using a mull prepared with a standard
absorbance of the clear aqueous solution of the substance
mixture obtained by mixing together thoroughly 1 part by
under examination.
weight of chloramphenicol palmitate (polymorph A) RS and
Sulphated ash (2.3.18). Not more than 0.1 per cent. 9 parts by weight of chloramphenicol palmitate RS. On each
283
CHLORAMPHENICOL SODIUM SUCCINATE IP 2007
of the spectra, draw a straight base line between the minima Description. A white or yellowish-white powder; hygroscopic.
occurring at about 880 cm–1 and 790 cm–1 and using these base
lines measure the heights of the peaks occuring at the maxima Identification
at about 858 cm–1 and 840 cm–1. In the spectrum obtained with A. Determine by thin-layer chromatography (2.4.17), coating
preparation under examination, the ratio of the peak height at the plate with silica gel GF254.
about 858 cm–1 to that at the maximum at about 840 cm–1 is
greater than the corresponding ratio in the spectrum obtained Mobile phase. A mixture of 85 volumes of chloroform,
with the standard mixture. 14 volumes of methanol and 1 volume of 2 M acetic acid.
Other tests. Complies with the tests stated under Oral Liquids. Test solution. Dissolve 0.1 g of the substance under
examination in 10 ml of acetone.
Assay. Weigh accurately a quantity of the suspension
containing about 125 mg of chloramphenicol, add 10 ml of Reference solution (a). A 1 per cent w/v solution of
water and shake with four quantities, each of 20 ml, of chloramphenicol sodium succinate RS in acetone.
chloroform, filtering each extract through cotton wool, Reference solution (b). A 1 per cent w/v solution of
previously washed with chloroform, into a 100-ml volumetric chloramphenicol RS in acetone.
flask. Dilute to volume with chloroform and mix well. Dilute
2.0 ml of this solution to 100.0 with ethanol (95 per cent) and
measure the absorbance of the resulting solution at the
The two principal spots in the chromatogram obtained with
maximum at about 271 nm using 1 ml of chloroform diluted to
the test solution are similar in position and size to those in the
50 ml with ethanol (95 per cent) as the blank (2.4.7). Calculate
chromatogram obtained with reference solution (a) and their
the content of chloramphenicol palmitate, C27H42Cl2N2O6,
positions are different from that of the principal spot in the
chromatogram obtained with reference solution (b).
Determine the weight per ml of the suspension (2.4.29) and
B. Dissolve 10 mg in 2 ml of ethanol (95 per cent) add 4.5 ml
calculate the content of chloramphenicol, C11H12Cl2N2O5,
of 1 M sulphuric acid and 50 mg of zinc powder, allow to
weight in volume using a factor of 0.575 for the conversion of
stand for 10 minutes and decant the supernatant liquid or
the content of chloramphenicol palmitate to chloramphenicol.
filter, if necessary. Cool the resulting solution in ice and add
Storage. Store protected from light. 0.5 ml of sodium nitrite solution and, after 2 minutes, 1 g of
Labelling. The label states (1) the strength in terms of the urea followed by 1 ml of 2-naphthol solution and 2 ml of 10 M
equivalent amount of chloramphenicol; (2) that if the sodium hydroxide; a red colour develops. Repeat the test
preparation is diluted, it must be used immediately after omitting the zinc powder; no red colour is produced.
dilution. C. To 5 ml of a 0.1 per cent w/v solution add a few drops of
silver nitrate solution; no precipitate is produced. Heat 50 mg
with 2 ml of ethanolic potassium hydroxide solution on a
Chloramphenicol Sodium Succinate water-bath for 15 minutes, add 50 mg of decolorising charcoal,
shake and filter. The filtrate when treated with silver nitrate
OH H Cl solution, yields a curdy precipitate which is insoluble in nitric
N acid but soluble, after being well washed with water, in dilute
Cl ammonia solution from which it is reprecipitated on addition
O of nitric acid.
O 2N O
D. A 5 per cent w/v solution gives the reactions of sodium
O COONa salts (2.3.1).
C15H15Cl2N2NaO8 Mol. Wt. 445.2 Tests
Chloramphenicol Sodium Succinate is a mixture of variable pH (2.4.24). 6.4 to 7.0, determined in a 25.0 per cent w/v solution.
proportions of sodium (2R,3R)-2-(2,2-dichloroacetamido)-3-
hydroxy-3-(4-nitrophenyl)propyl succinate (3-isomer) and of Specific optical rotation (2.4.22). +5.0° to +8.0°, determined in
sodium (1R,2R)-2-(2,2-dichloroacetamido)-3-hydroxy-1-(4- a 5.0 per cent w/v solution.
nitrophenyl)propyl succinate (1-isomer). Free chloramphenicol. Determine by thin-layer chromato-
Chloramphenicol Sodium Succinate contains not less than graphy (2.4.17), coating the plate with silica gel GF254.
98.0 per cent and not more than 102.0 per cent of Mobile phase. A mixture of 90 volumes of chloroform,
C15H15Cl2N2NaO8, calculated on the anhydrous basis. 10 volumes of methanol and 1 volume of water.
284
IP 2007 CHLORAMPHENICOL SODIUM SUCCINATE INJECTION
Test solution. Dissolve 0.1 g of the substance under Storage. The constituted solution should be used immediately
examination in 10 ml of acetone. after preparation but, in any case, within the period
Reference solution. A 0.02 per cent w/v solution of recommended by the manufacturer.
chloramphenicol RS in acetone. Chloramphenicol Sodium Succinate Injection contains not less
Apply to the plate 10 µl of each solution. After development, than 90.0 per cent and not more than 110.0 per cent of the
dry the plate in air and examine in ultraviolet light at 254 nm. stated amount of chloramphenicol, C11H12Cl2N2O5.
Any spot corresponding to chloramphenicol in the Description. A white or yellowish-white powder; hygroscopic.
intense than the spot in the chromatogram obtained with the The contents of the sealed container comply with the
reference solution. requirements stated under Parenteral Preparations
Assay. Weigh accurately about 0.2 g and dissolve in sufficient Identification
water to produce 500.0 ml; dilute 5.0 ml of this solution to A. Determine by thin-layer chromatography (2.4.17), coating
100.0 ml with water and measure the absorbance of the resulting the plate with silica gel GF254.
solution at the maximum at about 276 nm (2.4.7). Calculate the
content of C 15H15Cl2N 2NaO8 taking 220 as the specific Mobile phase. A mixture of 85 volumes of chloroform,
absorbance at 276 nm. 14 volumes of methanol and 1 volume of 2 M acetic acid.
Chloramphenicol Sodium Succinate intended for use in the Test solution. Dissolve 0.1 g of the substance under
manufacture of parenteral preparations without a further examination in 10 ml of acetone.
appropriate procedure for the removal of bacterial Reference Solution (a). A 1 per cent w/v solution of
endotoxins complies with the following additional chloramphenicol sodium succinate RS in acetone.
requirement.
Reference solution (b). A 1 per cent w/v solution of
Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit chloramphenicol RS in acetone.
per mg of chloramphenicol.
Chloramphenicol Sodium Succinate intended for use in the
The two principal spots in the chromatogram obtained with
the test solution are similar in position and size to those in the
chromatogram obtained with reference solution (a) and their
Sterility (2.2.11). Complies with the test for sterility. positions are different from that of the principal spot in the
Storage. Store protected from light and moisture. If the material chromatogram obtained with reference solution (b).
is intended for use in the manufacture of parenteral B. Dissolve 10 mg in 2 ml of ethanol (95 per cent) add 4.5 ml
preparations, the container should be sterile, tamper-evident of 1 M sulphuric acid and 50 mg of zinc powder, allow to
and sealed so as to exclude micro-organisms. stand for 10 minutes and decant the supernatant liquid or
Labelling. The label states whether or not the material is filter, if necessary. Cool the resulting solution in ice and add
intended for use in the manufacture of parenteral preparations. 0.5 ml of sodium nitrite solution and, after 2 minutes, 1 g of
urea followed by 1 ml of 2-naphthol solution and 2 ml of 10 M
sodium hydroxide; a red colour develops. Repeat the test
Chloramphenicol Sodium Succinate omitting the zinc powder; no red colour is produced.
Injection C. To 5 ml of a 0.1 per cent w/v solution add a few drops of
silver nitrate solution; no precipitate is produced. Heat 50 mg
Chloramphenicol Sodium Succinate Injection is a sterile with 2 ml of ethanolic potassium hydroxide solution on a
material consisting of Chloramphenicol Sodium Succinate with water-bath for 15 minutes, add 50 mg of decolorising charcoal,
or without excipients. It is filled in a sealed container. shake and filter. The filtrate when treated with silver nitrate
The injection is constituted by dissolving the contents of the solution, yields a curdy precipitate which is insoluble in nitric
sealed container in the requisite amount of sterile Water for acid but soluble, after being well washed with water, in dilute
Injections, immediately before use. ammonia solution from which it is reprecipitated on addition
of nitric acid.
Clarity of solution and Particulate matter stated under D. A 5 per cent w/v solution gives the reactions of sodium
Parenteral Preparations (Injections). salts (2.3.1).
285
CHLORBUTOL IP 2007
Tests Chlorbutol is 1,1,1-trichloro-2-methylpropan-2-ol

hemihydrate.
Chlorbutol contains not less than 98.0 per cent and not more
Specific optical rotation (2.4.22). +5.0° to +8.0°, determined in than 101.0 per cent of C4H7Cl3O, calculated on the anhydrous
a 5.0 per cent w/v solution. basis.
Free chloramphenicol. Determine by thin-layer Description. Colourless crystals or a white, crystalline powder;
chromatography (2.4.17), coating the plate with silica gel odour, characteristic and somewhat camphoraceous; sublimes
GF254. readily.
Mobile phase. A mixture of 90 volumes of chloroform,
10 volumes of methanol and 1 volume of water. Identification
Test solution. Dissolve 0.1 g of the substance under A. To 5 ml of a freshly prepared 0.5 per cent w/v solution add
examination and dissolve in 10 ml of acetone. 1 ml of 1 M sodium hydroxide and then, slowly, 2 ml of iodine
Reference solution. A 0.02 per cent w/v solution of solution; a yellow precipitate of iodoform is produced.
chloramphenicol RS in acetone. B. Heat about 20 mg with 2 ml of 10 M sodium hydroxide and
Apply to the plate 10 µl of each solution. After development, 1 ml of pyridine on a water-bath and shake; the separated
dry the plate in air and examine in ultraviolet light at 254 nm. pyridine layer becomes red.
Any spot corresponding to chloramphenicol in the C. Warm gently about 20 mg with 5 ml of ammoniacal silver
chromatogram obtained with the test solution is not more nitrate solution; a black precipitate is produced.
reference solution. Tests
Water (2.3.43). Not more than 2.0 per cent, determined on Appearance of solution. A 50.0 per cent w/v solution in ethanol
0.3 g. (95 per cent) is not more opalescent than opalescence
Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit standard OS2 (2.4.1), and not more intensely coloured than
per mg of chloramphenicol. reference solution BYS5 (2.4.1).
Sterility (2.2.11). Complies with the test for sterility. Acidity. Dissolve 2.0 g in 20 ml of ethanol (95 per cent), add
0.1 ml of bromothymol blue solution and titrate with 0.1 M
Assay. Determine the weight of the contents of 10 containers. sodium hydroxide; not more than 0.1 ml of 0.1 M sodium
Weigh accurately about 0.2 g of the mixed contents of the hydroxide is required to change the colour of the solution.
10 containers and dissolve in sufficient water to produce
500.0 ml; dilute 5.0 ml of this solution to 100.0 ml with water Chlorides (2.3.12). 0.5 g dissolved in 10 ml of ethanol (95 per
and measure the absorbance of the resulting solution at the cent) complies with the limit test for chlorides (500 ppm). Use
maximum at about 276 nm (2.4.7). Calculate the content of 5 ml of ethanol (95 per cent) in place of 5 ml of water to
C15H15Cl2N2NaO8 taking 220 as the specific absorbance at prepare the standard.
276 nm. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Storage. Store protected from light and moisture. Water (2.3.43). 4.5 per cent to 6.0 per cent, determined on
Labelling. The label states the quantity of Chloramphenicol 0.3 g.
Sodium Succinate in the sealed container in terms of the Assay. Weigh accurately about 0.2 g and dissolve in 5 ml of
equivalent amount of chloramphenicol. ethanol (95 per cent). Add 5 ml of sodium hydroxide solution
and boil under a reflux condenser for 15 minutes. Cool, dilute
with 20 ml of water, add 5 ml of nitric acid, 1 ml of nitrobenzene
and 50.0 ml of 0.1 M silver nitrate and shake vigorously for
Chlorbutol 1 minute. Add 4 ml of ferric ammonium sulphate solution and
titrate the excess of silver nitrate with 0.1 M ammonium
OH thiocyanate.
1 1 ml of 0.1 M silver nitrate is equivalent to 0.005917 g of
H3C CCl3 , /2H2O
C4H7Cl3O.
CH3
C4H7Cl3O, ½ H2O Mol.Wt. 186.5 exceeding 30°.
286
IP 2007 CHLORDIAZEPOXIDE
Chlorcyclizine Hydrochloride Test solution (b). A 0.1 per cent w/v solution of the substance
under examination in methanol.
Cl Reference solution (a). A 0.10 per cent w/v solution of
chlorcyclizine hydrochloride RS in methanol.
N , HCl methylpiperzine RS in methanol.
N substance under examination in methane.
CH3 Reference solution (d). 0.10 per cent w/v each of hydroxyzine
hydrochloride RS and chlorcyclizine hydrochloride RS in
C18H21ClN2, 2HCl Mol. Wt. 337.3 methanol.
Chlorcyclizine Hydrochloride is 1-(4-chlorobenzhydryl)-4- Apply to the plate 10 µl of each solution. After development,
methylpiperazine hydrochloride. dry the plate in air and expose to iodine vapours for 10 minutes.
Chlorcyclizine Hydrochloride contains not less than 99.0 per Any secondary spot in the chromatogram obtained with the
cent and not more than 101.0 per cent of the stated amount of test solution (a) is not more intense than the corresponding
C18H21CIN2,HCI, calculated on the dried basis. spot in the chromatogram obtained with the test solution (b).
Any spot other than the principle spot but corresponding to
Description. A white crystalline powder. the spot obtained with reference solution (a) is not more intense
Identification solution (c) (0.50 per cent). The test is not valid unless the
Test A may be omitted if tests B, C and D are carried out. Tests chromatogram obtained with reference solution (d) shows to
B and C may be omitted if tests A and D are carried out. clearly separated spot.
A. Determine by infrared absorption spectrophotometry (2.4.6). Sulphated ash (2.3.18). Not more than 0.1 per cent.
Compare the spectrum with that obtained with chlorcyclizine Loss on drying (2.4.19). Not more than 1.0 per cent, determined
hydrochloride RS or with the reference spectrum of on 1.0 g by drying in an oven at 130º.
chlorcyclizine hydrochloride.
Assay. Weigh accurately about 0.2 g, dissolve in 1 ml of 0.1 M
B. Weigh accurately about 10 mg, dissolve in 100 ml of 0.5 per hydrochloric acid and add 50 ml of methanol. Titrate with 0.1
cent w/v of suphuric acid. Dilute 10 ml of the solution to 100 M sodium hydroxide, determining the end-point
ml with 0.5 per cent w/v sulphuric acid. When examined in the potentiometrically (2.4.25). Carry out a blank titration.
range 215 to 300 nm (2.4.7), exhibits maximum only at about
231 nm; absorbance at about 231 nm, about 0.475 to 0.525. 1 ml of 0.1 M sodium hydroxide is equivalent to 0.03373 g of
C18H21CIN2,HCI.
C. In the test for Related substances, the principle spot in the
chromatogram obtained with test solution (b) corresponds to Storage. Store protected from light and moisture.
Chlordiazepoxide
Tests
Appearance of solution. A 5.0 per cent w/v solution is clear NHCH3
N
(2.4.1), and colourless (2.4.1).
Cl N
Related substance. Determine by thin layer chromatography O
(2.4.17). coating the plate with silica gel.
Mobile phase. A mixture of 85 volumes of dichloromethane,
solution. C16H14ClN3O MOl. Wt. 299.8
Test solution (a). A 2.0 per cent w/v solution of the substance Chlordiazepoxide is 7-chloro-2-methylamino-5-phenyl- 3H-
under examination in methanol. 1,4-benzodiazepine 4-oxide.
287
CHLORDIAZEPOXIDE TABLETS IP 2007
Chlordiazepoxide contains not less than 99.0 per cent and not intense than the spot in the chromatogram obtained with
more than 101.0 per cent of C16H14ClN3O, calculated on the reference solution (b). Spray the plate with a freshly prepared
dried basis. 1 per cent w/v solution of sodium nitrite in 1 M hydrochloric
acid, dry it in a current of air and spray with 0.4 per cent w/v
Description. An almost white to light yellow, crystalline
solution of N-(1-naphthyl)ethylenediamine dihydrochloride
powder; practically odourless.
in ethanol (95 per cent). Any violet spot in the chromatogram
obtained with the test solution corresponding to 2-amino-5-
Identification chlorobenzophenone is not more intense than the spot in the
Test A may be omitted if tests B and C are carried out. Tests B chromatogram obtained with reference solution (c).
and C may be omitted if test A is carried out. Heavy metals (2.3.13). 1.0 g complies with the limit test for
A. Determine by infrared absorption spectrophotometry (2.4.6). heavy metals, Method B (20 ppm).
Compare the spectrum with that obtained with Sulphated ash (2.3.18). Not more than 0.1 per cent.
chlordiazepoxide RS.
B. When examined in the range 230 nm to 360 nm (2.4.7) a on 1.0 g by drying in an oven at 105°.
0.0005 per cent w/v solution prepared immediately before use
in subdued light in 0.1 M hydrochloric acid shows absorption Assay. Weigh accurately about 0.25 g and dissolve by heating,
maxima at about 246 nm and 308 nm. Absorbance at the if necessary, in 80 ml of anhydrous glacial acetic acid. Titrate
maximum at about 246 nm, 0.56 to 0.60 and at the maximum at with 0.1 M perchloric acid, determining the end-point
about 308 nm, 0.16 to 0.17. potentiometrically (2.4.25). Carry out a blank titration.
C. Dissolve 0.2 g in 4 ml of hot dilute hydrochloric acid, heat 1 ml of 0.1 M perchloric acid is equivalent to 0.02998 g of
at 100° for 10 minutes, cool and filter. 2 ml of the filtrate gives C16H14ClN3O.
the reactions of primary aromatic amines (2.3.1). Storage. Store protected from light and moisture.
Tests
Related substances. Determine in subdued light by thin-layer
chromatography (2.4.17), coating the plate with silica gel Chlordiazepoxide Tablets
GF254. Chlordiazepoxide Tablets contain not less than 90.0 per cent
Mobile phase. A mixture of 70 volumes of toluene, 15 volumes and not more than 110.0 per cent of the stated amount of
of ethyl acetate, 10 volumes of ethanol (95 per cent), chlordiazepoxide, C16H14ClN3O. The tablets may be coated.
4 volumes of diethylamine and 1 volume of water.
Identification
A. Dilute 1 ml of the final solution obtained in the Assay to
Test solution. Dissolve 0.2 g in 10 ml of a mixture of 12 volumes
2 ml with 0.1 M hydrochloric acid. When examined in the
of methanol and 8 volumes of toluene.
range 230 nm to 360 nm (2.4.7) the resulting solution shows
Reference solution (a). Dilute 1 ml of the test solution to 10 ml absorption maxima at about 246 nm and 308 nm.
B. To a quantity of the powdered tablets containing 0.2 g of
Reference solution (b). Dilute 5 ml of reference solution (a) Chlordiazepoxide add 4 ml of hot 2 M hydrochloric acid, heat
to 100 ml with the same solvent mixture. at 100° for 10 minutes, cool and filter; 2 ml of the filtrate gives
Reference solution (c). A 0.005 per cent w/v solution of the reactions of primary aromatic amines (2.3.1).
2-amino-5-chlorobenzophenone in the same solvent mixture.
Tests
Reference solution (d). A 0.2 per cent w/v solution of
chlordiazepoxide RS in the same solvent mixture. Related substances. Determine by thin-layer chromatography
Apply to the plate 25 µl of the test solution as five quantities,
each of 5 µl, at one point, allowing the solvent to evaporate Mobile phase. A mixture of 85 volumes of chloroform,
between applications, and 5 µl of each of reference solutions 14 volumes of methanol and 1 volume of strong ammonia
(a), (b), (c) and (d). After development dry the plate in air and solution.
examine in ultraviolet light at 254 nm. Any secondary spot in Test solution. Shake a quantity of the powdered tablets
the chromatogram obtained with the test solution is not more containing 0.1 g of Chlordiazepoxide with 10 ml of a mixture of
288
IP 2007 CHLORHEXIDINE GLUCONATE SOLUTION
acetone containing 2 per cent v/v of strong ammonia solution Chlorhexidine Gluconate Solution
and 8 per cent v/v of water, allow to settle and use the clear
supernatant liquid. Chlorhexidine Gluconate Solution is an aqueous solution of
1,1’-hexamethylenebis [5-(4-chlorophenyl)biguanide]
Reference solution (a). Dilute 5 volumes of the test solution digluconate.
to 100 volumes with the same solvent mixture.
Chlorhexidine Gluconate Solution contains not less than
Reference solution (b). Dilute 1 volume of the test solution to 19.0 per cent w/v and not more than 21.0 per cent w/v of
100 volumes with the same solvent mixture. C22H30Cl2N10,2C6H12O7.
Reference solution (c). A 0.001 per cent w/v solution of Description. An almost colourless or pale yellowish, clear or
2-amino-5-chlorobenzophenone. slightly opalescent liquid; almost odourless.
Apply to the plate 2 µl and 20 µl quantities of the test solution,
2 µl of each of reference solutions (a) and (b) and 20 µl of Identification
reference solution (c). After development, dry the plate in air Test A may be omitted if tests B, C and D are carried out. Tests
and examine in ultraviolet light at 254 nm. Any secondary spot C and D may be omitted if tests A and B are carried out
in the chromatogram obtained with 2 µl of the test solution is
not more intense than the spot in the chromatogram obtained A. To 2 ml add 80 ml of water, cool in ice, add 5 M sodium
with reference solution (a) and not more than one such spot is hydroxide dropwise with stirring until the solution is slightly
more intense than the spot in the chromatogram obtained alkaline to titan yellow paper and add 2 ml in excess. Filter,
with reference solution (b). Spray the plate with a freshly wash the precipitate with water until the washings are free
prepared 1 per cent w/v solution of sodium nitrite in 1 M from alkali, dissolve it in about 25 ml of ethanol on a boiling
hydrochloric acid, dry it in a current of air and spray with a water-bath and heat until the volume is reduced to about 5 ml.
0.4 per cent w/v solution of N-(1-naphthyl)ethylenediamine Cool in ice, induce crystallisation, if necessary, by scratching
dihydrochloride in ethanol (95 per cent). Any violet spot the side of the vessel with a glass rod, filter and dry the crystals
corresponding to 2-amino-5-chlorobenzophenone in the at 105°. The residue complies with the following test.
chromatogram obtained with 20 µl of the test solution is not Determine by infrared absorption spectrophotometry (2.4.6).
more intense than the spot in the chromatogram obtained Compare the spectrum with that obtained with chlorhexidine
with reference solution (c). RS or with the reference spectrum of chlorhexidine. Examine
the substance as a dispersion in potassium bromide IR
without excessive grinding.
Tablets.
Powder one tablet, shake with 50 ml of 0.1 M hydrochloric
acid for 20 minutes and add sufficient 0.1 M hydrochloric
acid to produce 100.0 ml. Filter and dilute a suitable volume of Mobile phase. A mixture of 50 volumes of ethanol (95 per
the filtrate containing 0.8 mg of Chlordiazepoxide with sufficient cent), 30 volumes of water, 10 volumes of strong ammonia
0.1 M hydrochloric acid to produce 50.0 ml. Measure the solution and 10 volumes of ethyl acetate.
absorbance of the resulting solution at the maximum at about Test solution. Dilute 10 ml of the substance under examination
308 nm (2.4.7). Calculate the content of C16H14ClN3O in the to 50 ml with water.
tablet taking 327 as the specific absorbance at 308 nm.
Reference solution. A 2.5 per cent w/v solution of calcium
Other tests. Complies with the tests stated under Tablets. gluconate RS in water.
Assay. Weigh and powder 20 tablets. Weigh accurately a Apply to the plate 5 µl of each solution. Allow the mobile
quantity of the powder containing about 20 mg of phase to rise 10 cm. Dry the plate at 100° for 20 minutes, allow
Chlordiazepoxide and shake with 150 ml of 0.1 M hydrochloric to cool, spray with a 5 per cent w/v solution of potassium
acid for 20 minutes. Add sufficient 0.1 M hydrochloric acid dichromate in a 40 per cent w/w solution of sulphuric acid
to produce 250.0 ml and filter. Dilute 10.0 ml of the filtrate to and allow to stand for 5 minutes. The principal spot in the
50.0 ml with 0.1 M hydrochloric acid and measure the chromatogram obtained with the test solution corresponds to
absorbance of the resulting solution at the maximum at about that in the chromatogram obtained with the reference
308 nm (2.4.7). Calculate the content of C16H14ClN3O taking solution.
C. To 0.5 ml add 10 ml of water and 0.5 ml of cupric sulphate
Storage. Store protected from light at a temperature not solution; a white precipitate is produced which on boiling
exceeding 30°. flocculates and changes to a pale purple colour.
289
CHLORHEXIDINE ACETATE IP 2007
D. To 0.05 ml add 5 ml of a 1 per cent w/v solution of cetrimide, Titrate with 0.1 M perchloric acid, determining the end-point
1 ml of 10 M sodium hydroxide and 1 ml of bromine water; a potentiometrically (2.4.25). Carry out a blank titration.
deep red colour is produced. 1 ml of 0.1 M perchloric acid is equivalent to 0.02244 g of
Tests C22H30Cl2N10,2C6H12O7.
Determine the weight per ml (2.4.29) and calculate the
pH (2.4.24). 5.5 to 7.0, determined in a solution obtained by
percentage content of C22H30Cl2N10,2C6H12O7, weight in
diluting 5 ml to 100 ml.
volume.
Weight per ml (2.4.29). 1.06 g to 1.07 g, determined at 20°.
(2.4.17), coating a 0.5-mm thick plate with a slurry consisting
of 8 g of silica gel GF254 and 16 ml of water containing 1 g of
sodium formate. Chlorhexidine Acetate
50 volumes of ethanol (95 per cent) and 7 volumes of formic H H H
N N N
acid.
Test solution. Dilute 1 ml of the substance under examination NH NH
to 20 ml with 1.5 M acetic acid. Cl
, 2 H3 C CO2 H
Cl
Apply to the plate, in the form of a band 4 cm wide, 20 µl of the NH NH
test solution. After development, dry the plate in air and examine
in ultraviolet light at 254 nm. Mark the area around each group N N N
H H H
of bands above and below the principal band, transfer
quantitatively the enclosed areas of silica gel to a glass-
stoppered tube, add 5.0 ml of methanol, shake for 15 minutes, C22H30Cl2N10,2C2H4O2 Mol. Wt. 625.6
centrifuge and measure the absorbance of the clear, supernatant Chlorhexidine Acetate is 1,1'-(hexane-1,6-diyl)bis[5-(4-
liquid at the maximum at about 256 nm (2.4.7), using as the chlorophenyl)biguanide] diacetate.
blank a solution prepared by heating in a similar manner
Chlorhexidine Acetate contains not less than 98.0 per cent
equivalent-sized areas of silica gel removed from the coating
and not more than 101.0 per cent of chlorhexidine diacetate,
adjacent to the areas previously removed. The absorbance is
C22H30Cl2N10,2C2H4O2, calculated on the dried basis.
not more than that obtained with a solution prepared by
diluting 2 ml of the substance under examination with sufficient Description. A white or almost white, microcrystalline powder.
1.5 M acetic acid to produce 10 ml and diluting 0.2 ml of this
solution to 50 ml with methanol. Identification
4-Chloroaniline. Not more than 0.25 per cent, calculated with Test A may be omitted if tests B, C and D are carried out. Tests
reference to chlorhexidine solution at a nominal concentration B, C and D may be omitted if test A is carried out.
of 20 per cent w/v, determined by the following method. Dilute
2.0 ml to 100.0 with water. To 10.0 ml of this solution add 2.5 ml
Compare the spectrum with that obtained with chlorhexidine
of 2 M hydrochloric acid and dilute to 20 ml with water. Add
acetate RS.
rapidly, with continuous mixing after each addition, 0.35 ml of
sodium nitrite solution, 2 ml of a 5 per cent w/v solution of B. Dissolve about 5 mg in 5 ml of a warm 1.0 per cent w/v
ammonium sulphamate and 5 ml of a 0.01 per cent w/v solution solution of cetrimide and add 1 ml of strong sodium hydroxide
of N-(1-naphthyl) ethylenediamine dihydrochloride. Add solution and 1 ml of bromine water. A deep red colour is
1 ml of ethanol (95 per cent) and sufficient water to produce produced.
50 ml, mix and set aside for 30 minutes. Any reddish blue C. Dissolve 0.3 g in 10 ml of a mixture of equal volumes of
colour produced is not more intense than that produced by hydrochloric acid and water. Add 40 ml of water, filter if
treating at the same time in the same manner a mixture of necessary and cool in ice water. Make alkaline to titan yellow
10.0 ml of 0.001 per cent w/v solution of 4-chloroaniline in paper by adding dropwise and with stirring strong sodium
2 M hydrochloric acid and 10 ml of water in place of the hydroxide solution and add 1 ml in excess. Filter, wash the
dilution of the substance under examination. precipitate with water until the washings are free from alkali
Assay. Weigh accurately about 1.0 g and evaporate to a low and recrystallise from alcohol (70 per cent v/v). Dry at 100°
bulk. Dissolve in 50 ml of anhydrous glacial acetic acid. to 105°. Melting point (2.4.21). 132° to 136°.
290
IP 2007 CHLORHEXIDINE HYDROCHLORIDE
D. It gives reaction (a) of acetates (2.3.1). peaks, other than the principal peak is not greater than the
Tests reference solution (b) (2.5 per cent). Ignore any peak with a
relative retention time of 0.25 or less with respect to the
Chloroaniline. Dissolve 0.2 g of the substance under principal peak and any peak whose area is less than that of the
examination in 25 ml of water with shaking if necessary. Add 1 principal peak in the chromatogram obtained with reference
ml of hydrochloric acid and dilute to 30 ml with water. Add solution (c).
rapidly and with thorough mixing after each addition, 2.5 ml of
dilute hydrochloric acid, 0.35 ml of sodium nitrite solution, Loss on drying (2.4.19). Not more than 3.5 per cent, determined
2 ml of a 5.0 per cent w/v solution of ammonium sulphamate, on 1.0 g by drying in an oven at 105°.
5 ml of a 0.1 per cent w/v solution of naphthylethylenediamine Sulphated ash (2.3.18). Not more than 0.15 per cent.
dihydrochloride and 1 ml of alcohol, dilute to 50.0 ml with
Assay. Dissolve 0.14 g in 100 ml of anhydrous acetic acid and
water and allow to stand for 30 minutes. Any reddish-blue
titrate with 0.1 M perchloric acid determining the end-point
colour in the solution is not more intense than that in a standard
potentiometrically (2.4.25).
prepared at the same time in the same manner using a mixture
of 10.0 ml of 0.001 per cent w/v solution of chloroaniline in 1 ml of 0.1 M perchloric acid is equivalent to 0.01564 g of
dilute hydrochloric acid and 20 ml of dilute hydrochloric C26H38Cl2N10O4.
acid instead of the solution of the substance under examination
(500 ppm).
Related substances. Determine by liquid chromatography Chlorhexidine Hydrochloride
(2.4.14).
Test solution. Dissolve 0.2 g of the substance under H H H
examination in 100 ml of the mobile phase. N N N
NH NH
chlorhexidine acetate RS in the mobile phase. Cl
, 2 HCl
Reference solution (b). Dilute 2.5 ml of the test solution to Cl
NH NH
Reference solution (c) Dilute 2 ml of reference solution (b) to N N N
H H H
10 ml with the mobile phase. Further dilute 1 ml of this solution
C22H30Cl2N10,2HCl Mol. Wt. 578.4
– a stainless steel column 20 cm x 4 mm, packed with Chlorhexidine Hydrochloride is 1,1'-(hexane-1,6-diyl)bis[5-(4-
octadecylsilane bonded to porous silica (5 µm), chlorophenyl)biguanide] dihydrochloride.
– mobile phase: 2.0 g of sodium octanesulphonate in a Chlorhexidine Hydrochloride contains not less than
mixture of 120 ml of glacial acetic acid, 270 ml of water 98.0 per cent and not more than 101.0 per cent of chlorhexidine
and 730 ml of methanol, dihydrochloride, C22H30Cl2N10, 2HCl calculated on the dried
– flow rate. 1 ml per minute, basis.
– a 10 µl loop injector. Description. A white or almost white, crystalline powder.
Equilibrate the column with the mobile phase for at least Identification
1 hour. Adjust the sensitivity of the system so that the height
of the principal peak in the chromatogram obtained with Test A may be omitted if tests B, C and D are carried out. Tests
reference solution (b) is at least 50 per cent of the full scale of B, C and D may be omitted if test A is carried out.
the recorder. A. Determine by infrared absorption spectrophotometry (2.4.6).
Inject the test solution and reference solutions (a), (b) and (c). Compare the spectrum with that obtained with chlorhexidine
Record the chromatograms of reference solutions (b) and (c) hydrochloride RS.
until the peak due to chlorhexidine has been eluted and record B. Dissolve about 5 mg in 5 ml of a warm 1.0 per cent w/v
the chromatogram of the test solution for six times the retention solution of cetrimide and add 1 ml of strong sodium hydroxide
time of the peak due to chlorhexidine. In the chromatogram solution and 1 ml of bromine water. A deep red colour is
obtained with the test solution, the sum of the areas of all the produced.
291
CHLOROBUTANOL IP 2007
C. Dissolve 0.3 g in 10 ml of a mixture of equal volumes of reference solution (b) is at least 50 per cent of the full scale of
hydrochloric acid and water. Add 40 ml of water, filter if the recorder.
necessary and cool in ice water. Make alkaline to titan yellow Inject the test solution and reference solutions (a), (b) and (c).
paper by adding dropwise and with stirring strong sodium Record the chromatograms until the peak due to chlorhexidine
hydroxide solution and add 1 ml in excess. Filter, wash the has been eluted and record the chromatogram of the test
precipitate with water until the washings are free from alkali solution for six times the retention time of the peak due to
and recrystallise from alcohol (70 per cent v/v). Dry at 100° chlorhexidine. In the chromatogram obtained with the test
to 105°. Melting point (2.4.21). 132° to 136°. solution, the sum of the areas of all the peaks, other than the
D. It gives reaction (a) of chlorides (2.3.1). principal peak is not greater than the area of the principal peak
Tests (2.5 per cent). Ignore any peak with a relative retention time of
0.25 or less with respect to the principal peak and any peak
Chloroaniline. To 0.2 g of the substance under examination, whose area is less than that of the principal peak in the
add 1 ml of hydrochloric acid, dilute to 30 ml with water and chromatogram obtained with reference solution (c).
shake until a clear solution is obtained. Add rapidly and with
thorough mixing after each addition, 2.5 ml of dilute Loss on drying (2.4.19). Not more than 1.0 per cent, determined
hydrochloric acid, 0.35 ml of sodium nitrite solution, 2 ml of on 1.0 g by drying in an oven at 105°.
a 5.0 per cent w/v solution of ammonium sulphamate, 5 ml of Sulphated ash (2.3.18). Not more than 0.1 per cent.
a 0.1 per cent w/v solution of naphthylethylenediamine
Assay. Dissolve 0.1 g in 5 ml of anhydrous formic acid and
dihydrochloride and 1 ml of alcohol, dilute to 50.0 ml with
add 70 ml of acetic anhydride. Titrate with 0.1 M perchloric
water and allow to stand for 30 minutes. Any reddish-blue
acid determining the end-point potentiometrically (2.4.25).
colour in the solution is not more intense than that in a standard
prepared at the same time and in the same manner using a 1 ml of 0.1 M perchloric acid is equivalent to 0.01446 g of
mixture of 10.0 ml of a 0.001 per cent solution of chloroaniline C22H32Cl4N10.
in dilute hydrochloric acid and 20 ml of dilute hydrochloric
acid instead of the solution of the substance under examination
(500 ppm).
Related substances. Determine by liquid chromatography Chlorobutanol
(2.4.14).
Test solution. Dissolve 0.2 g of the substance under H3C OH
examination in 100 ml of the mobile phase. , ½ H2O
H3C CCl3
chlorhexidine hydrochloride RS in the mobile phase.
C4H7Cl3O,½H2O Mol. Wt. 186.5
Reference solution (b). Dilute 2.5 ml of the test solution to
100 ml with the mobile phase. Chlorobutanol is 1,1,1-trichloro-2-methylpropan-2-ol.
Reference solution (c). Dilute 2 ml of reference solution (b) to Chlorobutanol contains not less than 98.0 per cent and not
10 ml with the mobile phase. Dilute 1 ml of the solution to more than 101.0 per cent of chlorobutanol, C4H7Cl3O, calculated
10 ml with the mobile phase. on the anhydrous basis.
Chromatographic system Description. A white crystalline powder or colourless crystals.

octadecylsilane bonded to porous silica (5 µm), Identification
– mobile phase: 2.0 g of sodium octanesulphonate in a A. Dissolve 20 mg in a mixture of 1 ml of pyridine and 2 ml of
mixture of 120 ml of glacial acetic acid, 270 ml of water strong sodium hydroxide solution. Heat in a water-bath and
and 730 ml of methanol, shake. Allow to stand. The pyridine layer becomes red.
– spectrophotometer set at 254 nm, B. Dissolve 20 mg in 5 ml of ammoniacal silver nitrate solution
– a 10 µl loop injector. and warm slightly. A black precipitate is formed.
Equilibrate the column with the mobile phase for at least C. Dissolve 20 mg in 3 ml of 1 M sodium hydroxide. Add 5 ml
1 hour. Adjust the sensitivity of the system so that the height of water and then, slowly, 2 ml of iodinated potassium iodide
of the principal peak in the chromatogram obtained with solution. A yellowish precipitate is formed.
292
IP 2007 CHLOROCRESOL
Tests is produced, add 5 ml of water and filter. The melting range of

the residue, after crystallisation from methanol and drying at
Appearance of solution. A 50 per cent w/v solution in ethanol 70°, is 85° to 88° (2.4.21).
(95 per cent) (Solution A), is not more opalescent than
reference suspension II (2.4.1) and not more intensely coloured Tests
than reference solution BY5 ( 2.4.1).
Acidity. To 4 ml of solution A add 15 ml of ethanol (95 per Appearance of solution. A 5.0 per cent w/v solution in ethanol
cent) and 0.1 ml of bromothymol blue solution. Not more than (95 per cent) is clear (2.4.1) and not more intensely coloured
1.0 ml of 0.01 M sodium hydroxide is required to change the than reference solution BYS6 (2.4.1).
colour of the indicator to blue. Acidity or alkalinity. To 10 ml of a 5.0 per cent w/v solution
Chlorides (2.3.12). To 1 ml of solution A add 4 ml of ethanol add 0.1 ml of methyl red solution. The solution is orange or
(95 per cent) and dilute to 15 ml with water. The solution red and not more than 0.2 ml of 0.01 M sodium hydroxide is
complies with the limit test for chlorides (100 ppm). required to change the colour of the solution to yellow.
Water (2.3.43). 4.5 per cent to 5.5 per cent, determined on 0.3 g. Related substances. Determine by gas chromatography
(2.4.13).
Test solution. A 1 per cent w/v solution of the substance
Assay. Dissolve 0.1 g in 20 ml of ethanol (95 per cent), add 10 under examination in acetone.
ml of dilute sodium hydroxide solution, heat in a water-bath
for 5 minutes and cool. Add 20 ml of dilute nitric acid, 25.0 ml Chromatographic system
of 0.1 M silver nitrate and 2 ml of dibutyl phthalate and – a glass column 1.8m x 3.5 mm, packed with silanised
shake vigorously. Add 2 ml of ferric ammonium sulphate diatomaceous support (80 to 120 mesh) impregnated
solution and titrate with 0.1 M ammonium thiocyanate until with 3 to 5 per cent w/w of phenyl methyl silicone fluid
an orange colour is obtained. (50 per cent phenyl) (such as OV-17),
– temperature:
1 ml of 0.1 M silver nitrate is equivalent to 0.00592 g of
column.125°,
C4H7Cl3O.
inlet port. 210°,
Storage. Store protected from moisture. detector. 230° ,
– flow rate. 30 ml per minute of the carrier gas (nitrogen).
Allow the chromatography to proceed for three times the
Chlorocresol retention time of chlorocresol (about 8 minutes).
The sum of the areas of any secondary peaks in the
OH chromatogram is not greater than 1.0 per cent of the total area
of the peaks.
Non-volatile matter. Not more than 0.1 per cent, determined
CH3 on 2.0 g by volatilising on a water-bath and drying at 105°.
Cl Assay. Weigh accurately about 70 mg, dissolve in 30 ml of
C7H7ClO Mol. Wt. 142.6 glacial acetic acid, add 25.0 ml of 0.0167 M potassium
bromate, 20.0 ml of a 15 per cent w/v solution of potassium
Chlorocresol is 4-chloro-3-methylphenol.
bromide and 10 ml of hydrochloric acid. Stopper the flask
Chlorocresol contains not less than 98.0 per cent and not and allow to stand in the dark for 15 minutes, shaking
more than 101.0 per cent of C7H7ClO. occasionally. Add 1 g of potassium iodide and 100 ml of water.
Description. Colourless or almost colourless crystals or a Titrate with 0.1 M sodium thiosulphate, shaking vigorously
white, crystalline powder; odour, characteristic and not tarry; and using starch solution, added towards the end of the
volatile in steam. titration, as indicator. Repeat the procedure without the
Identification titrations represents the amount of potassium bromate
required.
A.To a saturated solution in water add one drop of ferric
chloride test solution; a bluish colour is produced. 1 ml of 0.0167 M potassium bromate is equivalent to
0.003565 g of C7H7ClO.
B.To 0.1 g add 0.2 ml of benzoyl chloride and 0.5 ml of 2 M
sodium hydroxide. Shake vigorously until a white precipitate Storage. Store protected from light and moisture.
293
CHLOROFORM IP 2007
Chloroform Shake 15 ml of the chloroform layer with 30 ml of water in a

stoppered flask for 3 minutes and allow to separate. To the
CHCl3 Mol. Wt. 119.4 aqueous layer add 0.2 ml of silver nitrate solution and set
Chloroform is trichloromethane to which either 1.0 per cent to aside in the dark for 5 minutes; no opalescence is produced.
2.0 per cent v/v of ethanol or 50 mg per litre of amylene has Related substances. Determine by gas chromatography
been added. (2.4.13).
Description. A colourless, volatile liquid; odour, characteristic. Test solution (a). A solution containing 0.2 per cent v/v of
NOTE - Care should be taken not to vaporise chloroform in carbon tetrachloride, 0.2 per cent v/v of 1,1,1-trichloroethane
the presence of a flame because of the production of harmful (internal standard), 0.2 per cent v/v of dichloromethane,
gases. 0.2 per cent v/v of ethanol, 0.5 per cent v/v of
bromochloromethane and 0.2 per cent v/v of the substance
Identification under examination in 1-propanol.
Test A may be omitted if tests B and C are carried out. Tests B Test solution (b). The substance under examination.
and C may be omitted if test A is carried out. Reference solution (a). A solution containing 0.2 per cent v/v
of the internal standard in the substance under examination.
A. Shake with an equal volume of water and dry with
anhydrous sodium sulphate. Determine by infrared absorption Reference solution (b). 1-propanol.
obtained with the reference spectrum of chloroform.
– a glass column 4m x 3 mm, packed with acid-washed
B. Non-flammable. The vapour introduced into a Bunsen flame kieselguhr (60 to 100 mesh) coated with 15 per cent w/w
produces a green colour and gives rise to noxious vapours of di-2-cyanoethyl ether,
having a characteristic odour. – temperature:
C. Warm 0.5 ml with 0.05 ml of aniline and 1 ml of 5 M sodium column.40°,
hydroxide. The characteristic odour of phenyl isocyanide is inlet port and detector. 100°,
produced. – flow rate. 30 ml per minute of the carrier gas.
– Inject 0.1 µl of each solution.
Tests The test is not valid unless the column efficiency, determined
using the chloroform peak in the chromatogram obtained with
Weight per ml (2.4.29). 1.474 g to 1.478 g. test solution (a), is greater than 700 plates per metre and the
Boiling range (2.4.8). Not more than 5.0 per cent v/v distils total number of plates is greater than 2,500.
below 60° and the remainder distils between 60° and 62°. In the chromatogram obtained with test solution (a) the peaks,
Acidity or alkalinity. Shake 10 ml with 20 ml of freshly boiled in the order of emergence, are due to carbon tetrachloride,
and cooled water for 3 minutes and allow to separate. To 5 ml 1,1,1-trichloroethane, dichloromethane, chloroform, ethanol,
of the aqueous layer (solution A) add 0.1 ml of litmus solution; bromochloromethane and 1-propanol (solvent).
the colour produced is similar to that produced on adding Using the chromatogram obtained with reference solution (b)
0.1 ml of litmus solution to 5 ml of freshly boiled and cooled make any corrections due to the contribution of secondary
water. peaks from the solvent to the peaks in the chromatogram
Chlorides. To 5 ml of solution A add 5 ml of water and 0.2 ml obtained with test solution (a).
of silver nitrate solution; the solution is clear. In the chromatogram obtained with reference solution (a), the
Free chlorine. To 10 ml of solution A add 1 ml of cadmium ratio of the areas of any peaks due to carbon tetrachloride,
iodide solution and 2 drops of starch solution; no blue colour dichloromethane and bromochloromethane to the area of the
is produced. peak due to the internal standard is not greater than the
corresponding ratios in the chromatogram obtained with test
Aldehyde. Shake 5 ml with 5 ml of water and 0.2 ml of alkaline
solution (a) and the ratio of the area of any other secondary
potassium mercuri-iodide solution in a stoppered bottle and
peak that elutes prior to the solvent peak, except for the peak
set aside in the dark for 15 minutes; not more than a pale
corresponding to ethanol, to the area of the peak due to the
yellow colour is produced.
internal standard is not greater than the ratio of the area of the
Foreign chlorine compounds. Shake 20 ml with 10 ml of peak due to chloroform to the area of the peak due to the
sulphuric acid in a stoppered flask for 5 minutes, allow to internal standard in the chromatogram obtained with test
stand in the dark for 30 minutes and discard the acid layer. solution (a).
294
IP 2007 CHLOROQUINE PHOSPHATE
Calculate the percentage content of each of the specified Chloroquine Phosphate contains not less than 98.5 per cent
impurities and also calculate the percentage content of each and not more than 101.0 per cent of C18H26ClN3,2H3PO4,
of any other impurities assuming the same response per unit calculated on the anhydrous basis.
volume as with chloroform. The total content of all impurities Description. A white or almost white, crystalline powder;
is not more than 1.0 per cent v/v. odourless. It slowly gets discoloured on exposure to light. It
Ethanol (if present). Determine by gas chromatography may exist in two polymorphic forms differing in their behaviour,
(2.4.13). one of which melts at about 195° and the other at about 218°.
Test solution (a). The substance under examination.
Identification
Test solution (b). A solution containing 1.0 per cent v/v of
1-propanol (internal standard) in the substance under Test A may be omitted if tests B, C and D are carried out. Tests
examination. B and C may be omitted if tests A and D are carried out.
Reference solution. A solution containing 1.0 per cent v/v of A. Dissolve 0.1 g in 10 ml of water, add 2 ml of 2 M sodium
ethanol and 1.0 per cent v/v of the internal standard in water. hydroxide and extract with two quantities, each of 20 ml, of
chloroform. Wash the combined chloroform extracts with water,
Inject 0.1 µl of each solution. dry over anhydrous sodium sulphate, evaporate to dryness
Follow the chromatographic procedure described under and dissolve the residue in 2 ml of chloroform. The resulting
Related substances. solution complies with the following test.
The test is not valid unless the height of the trough separating Determine by infrared absorption spectrophotometry (2.4.6).
the ethanol peak from the chloroform peak in the chromatogram Compare the spectrum with that obtained with 80 mg of
obtained with test solution (a) is less than 15 per cent of the chloroquine phosphate RS treated in the same manner.
height of the ethanol peak.
Calculate the percentage content of ethanol from the areas of 0.001 per cent w/v solution shows absorption maxima at about
the peaks due to ethanol and the internal standard in the 220 nm, 235 nm, 256 nm, 329 nm and 342 nm; absorbance at
chromatograms obtained with test solution (a) and test about 220 nm, 0.60 to 0.66, at about 235 nm, 0.35 to 0.39, at
solution (b). about 256 nm, 0.30 to 0.33, at about 329 nm, 0.325 to 0.355 and
Non-volatile matter. Not more than 0.004 per cent w/v, at about 342 nm, 0.36 to 0.39.
determined on 25 ml by evaporation to dryness and drying at C. Dissolve 25 mg in 20 ml of water and add 8 ml of picric acid
105°. solution; the precipitate, after washing successively with
Storage. Store protected from light in tightly-closed, glass- water, ethanol (95 per cent) and ether, melts at 205° to 210°
stoppered containers. (2.4.21).
Labelling. The label states whether it contains ethanol or D. Neutralise with dilute nitric acid the aqueous layer obtained
amylene. in test A. Add an equal volume of ammonium molybdate
solution and warm; a yellow precipitate is produced.
Tests
Chloroquine Phosphate Appearance of solution. A 10.0 per cent w/v solution in carbon
dioxide-free water is clear (2.4.1), and not more intensely
coloured than reference solution BYS5 or GYS5 (2.4.1).
Cl N
,2H3PO4 (2.4.17), coating the plate with silica gel GF254.
HN
N CH3
CH3
CH3 40 volumes of cyclohexane and 10 volumes of diethylamine.
C18H26ClN3,2H3PO4 Mol. Wt. 515.9 under examination in water.
Chloroquine Phosphate is (RS)-7-chloro-4-(4-diethylamino- Reference solution (a). Dilute 1 ml of the test solution to
1-methylbutylamino)quinoline diphosphate. 100 ml with water.
295
CHLOROQUINE PHOSPHATE INJECTION IP 2007
Reference solution (b). Dilute 25 ml of reference solution (a) C. Neutralise the aqueous layer obtained in test A with dilute
to 50 ml with water. nitric acid, add an equal volume of ammonium molybdate
Apply to the plate 2 µl of each solution. Allow the mobile solution and warm; a yellow precipitate is produced.
phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
Tests
light at 254 nm. Any secondary spot in the chromatogram
obtained with the test solution is not more intense than the pH (2.4.24). 3.5 to 4.5.
spot in the chromatogram obtained with reference solution (a)
and not more than one such spot is more intense than the
(b). Assay. To an accurately measured volume of the injection
Heavy metals (2.3.13). 2.0 g complies with the limit test for containing 0.4 g of chloroquine add 20 ml of 1 M sodium
heavy metals, Method A (10 ppm). hydroxide and extract with four quantities, each of 25 ml, of
chloroform. Combine the chloroform extracts and evaporate
Water (2.3.43). Not more than 2.0 per cent, determined on to a volume of about 10 ml. Add 40 ml of anhydrous glacial
1.0 g. acetic acid and mix. Titrate with 0.1 M perchloric acid,
Assay. Weigh accurately about 0.2 g and dissolve in 50 ml of determining the end-point potentiometrically (2.4.25). Carry
anhydrous glacial acetic acid with the aid of heat (if necessary, out a blank titration.
heat under a reflux condenser). Titrate with 0.1 M perchloric 1 ml of 0.1 M perchloric acid is equivalent to 0.01599 g of
acid, determining the end-point potentiometrically (2.4.25). chloroquine, C18H26ClN3.
C18H26ClN3,2H3PO4. Labelling. The label states the strength in terms of the
equivalent amount of chloroquine in a suitable dose-volume.
Chloroquine Phosphate Injection Chloroquine Phosphate Suspension

Chloroquine Phosphate Injection is a sterile solution of Chloroquine Phosphate Suspension is a suspension of
Chloroquine Phosphate in Water for Injections. Chloroquine Phosphate in a suitable flavoured vehicle.
Chloroquine Phosphate Injection contains not less than Chloroquine Phosphate Suspension contains not less than
95.0 per cent and not more than 105.0 per cent of the stated 95.0 per cent and not more than 105.0 per cent of the stated
amount of chloroquine, C18H26ClN3. amount of chloroquine, C18H26ClN3.
Description. A clear, colourless or almost colourless solution. Identification

Identification To a volume of the suspension containing 50 mg of
chloroquine add 2 ml of 2 M sodium hydroxide and extract
A. To a volume of the injection containing 60 mg of
with two quantities, each of 20 ml, of chloroform. Wash the
chloroquine add 2 ml of 2 M sodium hydroxide and extract
combined chloroform extracts with water, dry over anhydrous
with two quantities, each of 20 ml, of chloroform. Wash the
sodium sulphate, evaporate to dryness and dissolve the
combined chloroform extracts with water, dry over anhydrous
residue in 2 ml of chloroform. The resulting solution complies
sodium sulphate, evaporate to dryness and dissolve the
with the following test.
residue in 2 ml of chloroform. The resulting solution complies
with the following test. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with 80 mg of
chloroquine phosphate RS treated in the same manner.
Compare the spectrum with that obtained with 80 mg of
chloroquine phosphate RS treated in the same manner. Tests
B. Dilute a volume of the injection containing 15 mg of pH (2.4.24). 5.5 to 6.5.
chloroquine to 20 ml with water and add 8 ml of picric acid
solution; the precipitate, after washing successively with Other tests. Complies with the tests stated under Oral Liquids.
water, ethanol (95 per cent) and ether, melts at about 207° Assay. Weigh accurately a quantity of the suspension
(2.4.21). containing about 100 mg of chloroquine, add 50 ml of 1 M
296
IP 2007 CHLOROQUINE SULPHATE
hydrochloric acid, shake well and dilute to 100.0 ml with 1 M Reference solution (a). Dilute 1 ml of the test solution to
hydrochloric acid. Filter and discard the first few ml of the 100 ml with water.
filtrate. Dilute 10.0 ml of the filtrate to 100.0 ml with 1 M Reference solution (b). Dilute 25 ml of reference solution (a)
hydrochloric acid and mix. Further dilute 10.0 ml to 100.0 ml to 50 ml with water.
with the same solvent and mix. Measure the absorbance of
the resulting solution at the maximum at about 342 nm (2.4.7). Apply to the plate 2 µl of each solution. Allow the mobile
Calculate the content of C18H26ClN3 from the absorbance phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
obtained by repeating the operation using chloroquine light at 254 nm. Any secondary spot in the chromatogram
phosphate RS in place of the substance under examination. obtained with the test solution is not more intense than the
spot in the chromatogram obtained with reference solution (a)
and not more than one such spot is more intense than the
(b).
Chloroquine Phosphate Tablets
Chloroquine Phosphate Tablets contain not less than 92.5 per
cent and not more than 107.5 per cent of the stated amount of Apparatus. No 1
chloroquine phosphate, C18H26ClN3,2H3PO4 The tablets are Medium. 900 ml of 0.1 M hydrochloric acid
coated. Speed and time. 75 rpm and 45 minutes.
Identification through a membrane filter disc with an average pore diameter
A. To a quantity of the powdered tablets containing 0.1 g of
and dilute a suitable volume of the filtrate with 0.1 M
Chloroquine Phosphate add 10 ml of water and 2 ml of 2 M
hydrochloric acid. Measure the absorbance of the resulting
sodium hydroxide and extract with two quantities, each of
20 ml, of chloroform. Wash the combined chloroform extracts
content of C18H26ClN3,2H3PO4 per tablet taking 371 as the
with water, dry over anhydrous sodium sulphate, evaporate
specific absorbance at 344 nm.
to dryness and dissolve the residue in 2 ml of chloroform. The
resulting solution complies with the following test. D. Not less than 70 per cent of the stated amount of
Determine by infrared absorption spectrophotometry (2.4.6). C18H26ClN3,2H3PO4.
Compare the spectrum with that obtained with 80 mg of Other tests. Comply with the tests stated under Tablets.
chloroquine phosphate RS treated in the same manner.
B. Extract a quantity of the powdered tablets containing 25 mg quantity of the powder containing about 0.5 g of Chloroquine
of Chloroquine Phosphate with 20 ml of water, filter and to the Phosphate, add 20 ml of 1 M sodium hydroxide and extract
filtrate add 8 ml of picric acid solution; the precipitate, after with four quantities, each of 25 ml, of chloroform. Combine
washing successively with water, ethanol (95 per cent) and the chloroform extracts and evaporate to a volume of about
ether, melts at about 207° (2.4.21). 10 ml. Add 40 ml of anhydrous glacial acetic acid and mix.
Titrate with 0.1 M perchloric acid, determining the end-point
C. Extract a quantity of the powdered tablets containing 0.5 g
potentiometrically (2.4.25). Carry out a blank titration.
of Chloroquine Phosphate with 25 ml of water and filter. To
the filtrate add 2.5 ml of 5 M sodium hydroxide and extract 1 ml of 0.1 M perchloric acid is equivalent to 0.02579 g of
with three quantities, each of 10 ml, of ether. The aqueous C18H26ClN3,2H3PO4.
layer, after neutralisation with 2 M nitric acid, gives the Storage. Store protected from light.
reactions of phosphates (2.3.1).
Tests
Chloroquine Sulphate
(2.4.17), coating the plate with silica gel GF254. C18H26ClN3,H2SO4,H2O Mol. Wt. 435.9
Mobile phase. A mixture of 50 volumes of chloroform, Chloroquine Sulphate is (RS)-4-(7-chloro-4- quinolyl-
40 volumes of cyclohexane and 10 volumes of diethylamine. amino) pentyldiethylamine sulphate monohydrate.
Test solution. Shake a quantity of the powdered tablets Chloroquine Sulphate contains not less than 98.5 per cent
containing 1 g of Chloroquine Phosphate with 20 ml of water and not more than 101.0 per cent of C18H26ClN3,H2SO4,
for 30 minutes, centrifuge and use the clear, supernatant liquid. calculated on the anhydrous basis.
297
CHLOROQUINE SULPHATE INJECTION IP 2007
Description. A white or almost white, crystalline powder; spot in the chromatogram obtained with reference solution (a)
odourless. and not more than one such spot is more intense than the spot
in the chromatogram obtained with reference solution (b).
Identification Heavy metals (2.3.13). 1.0 g dissolved in 25 ml of water complies
Test A may be omitted if tests B, C and D are carried out. Tests with the limit test for heavy metals, Method A (20 ppm).
B and C may be omitted if tests A and D are carried out. Chlorides (2.3.12). 1.25 g complies with the limit test for
A. Dissolve 0.1 g in 10 ml of water, add 2 ml of 2 M sodium chlorides (200 ppm).
hydroxide and extract with two quantities, each of 20 ml, of Sulphated ash (2.3.18). Not more than 0.1 per cent.
chloroform. Wash the combined chloroform extracts with water,
dry with anhydrous sodium sulphate, evaporate to dryness
and dissolve the residue in 2 ml of chloroform. The resulting Assay. Weigh accurately about 0.5 g and dissolve in 50 ml of
solution complies with the following test. anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
Compare the spectrum with that obtained by treating 0.1 g of
chloroquine sulphate RS in the same manner. 1 ml of 0.1 M perchloric acid is equivalent to 0.0418 g of
C18H26ClN3,H2SO4.
B. When examined in the range 210 nm to 360 nm, a 0.001 per
cent w/v solution shows absorption maxima at about 220 nm, Storage. Store protected from light.
235 nm, 256 nm, 329 nm and 342 nm; absorbance at about
220 nm, 0.73 to 0.81, at about 235 nm, 0.43 to 0.47, at about
256 nm, 0.37 to 0.41, at about 329 nm, 0.40 to 0.44 and at about Chloroquine Sulphate Injection
342 nm, 0.43 to 0.47 (2.4.7).
Chloroquine Sulphate Injection is a sterile solution of
C. Dissolve 25 mg in 20 ml of water and add 8 ml of picric acid Chloroquine Sulphate in Water for Injections.
solution; the precipitate, after washing successively with
water, ethanol (95 per cent) and ether, melts at 205° to 210° Chloroquine Sulphate Injection contains not less than
(2.4.21). 95.0 per cent and not more than 105.0 per cent of the stated
amount of chloroquine, C18H26ClN3.
D. Gives reaction A of sulphates (2.3.1).
Description. A clear, colourless or almost colourless solution.
Tests
Identification
Appearance of solution. An 8.0 per cent w/v solution in carbon
A. To a volume of the injection containing 70 mg of chloroquine
add sufficient water to produce 10 ml, add 2 ml of 2 M sodium
coloured than reference solution BYS5 or GYS5 (2.4.1).
hydroxide and extract with two quantities, each of 20 ml, of
pH (2.4.24). 4.0 to 5.0, determined in an 8.0 per cent w/v chloroform. Wash the combined chloroform extracts with water,
solution. dry with anhydrous sodium sulphate, evaporate to dryness
Related substances. Determine by thin-layer chromatography and dissolve the residue in 2 ml of chloroform. The resulting
(2.4.17), coating the plate with silica gel GF254. solution complies with the following test.
Mobile phase. A mixture of 50 volumes of chloroform, Determine by infrared absorption spectrophotometry (2.4.6).
40 volumes of cyclohexane and 10 volumes of diethylamine. Compare the spectrum with that obtained by treating 0.1 g of
chloroquine sulphate RS in the same manner.
under examination in water. B. When examined in the range 210 nm to 360 nm, a 0.001 per
cent w/v solution shows absorption maxima at about 220 nm,
Reference solution (a). Dilute 1 ml of the test solution to 235 nm, 256 nm, 329 nm and 342 nm; absorbance at about
100 ml with water. 220 nm, 0.73 to 0.81, at about 235 nm, 0.43 to 0.47, at about
Reference solution (b). Dilute 25 ml of reference solution (a) 256 nm, 0.37 to 0.41, at about 329 nm, 0.40 to 0.44 and at about
to 50 ml with water. 342 nm, 0.43 to 0.47 (2.4.7).
Apply to the plate 2 µl of each solution. Allow the mobile C. Gives reaction A of sulphates (2.3.1).
Tests
obtained with the test solution is not more intense than the pH (2.4.24). 4.0 to 5.5.
298
IP 2007 CHLOROQUINE SYRUP
Other tests. Complies with the tests stated under Parenteral Speed and time. 75 rpm and 45 minutes.
Preparations (Injections). Withdraw a suitable volume of the medium and filter promptly
Assay. To an accurately measured volume of the injection through a membrane filter disc with an average pore diameter
containing 0.4 g of chloroquine add 20 ml of 1 M sodium not greater than 1.0 µm. Reject the first few ml of the filtrate
hydroxide and extract with four quantities, each of 25 ml, of and dilute a suitable volume of the filtrate with 0.1 M
chloroform. Combine the chloroform extracts and evaporate hydrochloric acid. Measure the absorbance of the resulting
to a volume of about 10 ml. Add 40 ml of anhydrous glacial solution at the maximum at about 344 nm (2.4.7). Calculate the
acetic acid and mix. Titrate with 0.1 M perchloric acid, content of C18H26ClN3,H2SO4 per tablet taking 450 as the
determining the end-point potentiometrically (2.4.25). Carry specific absorbance at 344 nm.
1 ml 0.1 M perchloric acid is equivalent to 0.01599 g of C18H26ClN3,H2SO4.
C18H26ClN3.
Storage. Store protected from light. Assay. Weigh and powder 20 tablets. Weigh accurately a
Labelling. The label states the strength in terms of the quantity of the powder containing about 0.5 g of Chloroquine
equivalent amount of chloroquine in a suitable dose-volume. Sulphate, add 20 ml of 1 M sodium hydroxide and extract with
four quantities, each of 25 ml, of chloroform. Combine the
chloroform extracts and evaporate to a volume of about 10 ml.
Add 40 ml of anhydrous glacial acetic acid and mix. Titrate
Chloroquine Sulphate Tablets with 0.1 M perchloric acid, determining the end-point
Chloroquine Sulphate Tablets contain not less than 92.5 per potentiometrically (2.4.25). Carry out a blank titration.
chloroquine sulphate, C18H26ClN3,H2SO4. The tablets are
C18H26ClN3,H2SO4.
coated.
Identification
A. To a quantity of the powdered tablets equivalent to 0.1 g of
Chloroquine Sulphate add 10 ml of water and 2 ml of 2 M
sodium hydroxide and extract with two quantities, each of 20 Chloroquine Syrup
ml, of chloroform. Wash the combined chloroform extracts Chloroquine Syrup is a solution of Chloroquine Phosphate or
with water, dry with anhydrous sodium sulphate, evaporate Chloroquine Sulphate in a suitable flavoured vehicle.
to dryness and dissolve the residue in 2 ml of chloroform. The
Chloroquine Syrup contains Chloroquine Phosphate or
resulting solution complies with the following test.
Chloroquine Sulphate equivalent to not less than 95.0 per
Determine by infrared absorption spectrophotometry (2.4.6). cent and not more than 105.0 per cent of the stated amount of
Compare the spectrum with that obtained by treating 0.1 g of chloroquine, C18H26ClN3.
B. Extract a quantity of the powdered tablets containing 25 mg Identification
of Chloroquine Sulphate with 20 ml of water, filter and to the To a volume of the syrup containing 50 mg of chloroquine add
filtrate add 8 ml of picric acid solution; the precipitate, after 2 ml of 2 M sodium hydroxide and extract with two quantities,
washing successively with water, ethanol (95 per cent) and each of 20 ml, of chloroform. Wash the combined chloroform
ether, melts at about 207° (2.4.21). extracts with water, dry with anhydrous sodium sulphate,
C. Extract a quantity of the powdered tablets containing about evaporate to dryness and dissolve the residue in 2 ml of
0.1 g of Chloroquine Sulphate with 10 ml of water and 1 ml of chloroform. The resulting solution complies with the following
dilute hydrochloric acid and filter. To the filtrate add 1 ml of test.
barium chloride solution; a white precipitate is produced. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained by treating 0.1 g of
Tests
Tests
Apparatus. No 1
Medium. 900 ml of 0.1 M hydrochloric acid pH (2.4.24). 4.0 to 6.5.
299
CHLOROXYLENOL IP 2007
Other tests. Complies with the tests stated under Oral Liquids. Tests
Assay. To an accurately measured volume of the syrup Related substances. Determine by gas chromatography
containing about 0.4 g of chloroquine add 20 ml of 1 M sodium (2.4.13).
hydroxide and extract with four quantities, each of 25 ml, of
chloroform. Combine the chloroform extracts and evaporate
under examination in chloroform.
to a volume of about 10 ml. Add 40 ml of anhydrous glacial
acetic acid and mix. Titrate with 0.1 M perchloric acid, Reference solution. A solution containing 2 per cent w/v of
determining the end-point potentiometrically (2.4.25). Carry the substance under examination and 0.04 per cent w/v of
out a blank titration. 4-chloro-o-cresol (internal standard) in chloroform.
1 ml of 0.1 M perchloric acid is equivalent to 0.01599 g of Chromatographic system
C18H26ClN3. – a glass column 1.5m × 4 mm, packed with acid-washed
diatomaceous support (80 to 100 mesh) coated with
3 per cent w/w of polyethylene glycol (such as Carbowax
Labelling. The label states (1) whether the syrup contains 20M),
Chloroquine Phosphate or Chloroquine Sulphate; (2) the – temperature:
strength in terms of equivalent amount of chloroquine in each column.160°,
5 ml. inlet port and detector. 220°,
– a flame ionisation detector,
In the chromatogram obtained with the reference solution the
Chloroxylenol sum of the areas of any secondary peaks is not greater than
the area of the peak due to internal standard.
OH Assay. Weigh accurately about 70 mg, dissolve in 30 ml of
glacial acetic acid, add 25.0 ml of 0.0167 M potassium
bromate, 20 ml of a 15 per cent w/v solution of potassium
H3C CH3 bromide and 10 ml of hydrochloric acid, stopper the flask
and allow to stand protected from light for 15 minutes. Add
Cl
1 g of potassium iodide and 100 ml of water and titrate with
C8H9ClO Mol. Wt. 156.6 0.1 M sodium thiosulphate, shaking vigorously and using
1 ml of starch solution as indicator. Repeat the procedure
Chloroxylenol is 4-chloro-3,5-dimethylphenol. without the substance under examination. The difference
Chloroxylenol contains not less than 98.0 per cent and not between the titrations represents the amount of potassium
more than 103.0 per cent of C8H9ClO. bromate required.
Description. A white or creamy-white crystals or crystalline 1 ml of 0.0167 M potassium bromate is equivalent to
powder; odour characteristic. It is volatile in steam. 0.003915 g of C8H9ClO.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). Chloroxylenol Solution
Compare the spectrum with that obtained with chloroxylenol Chloroxylenol solution is a solution of Chloroxylenol
RS or with the reference spectrum of chloroxylenol. solubilised in a saponaceous base containing Ethanol (95 per
B. Dissolve 0.1 g in 5 ml of chloroform and add 0.5 ml of a cent) and essential oils. Ethanol (95 per cent) may be replaced
filtered 1 per cent w/v solution of ferric chloride in chloroform by Industrial Methylated Spirit in making Chloroxylenol
and 0.1 ml of pyridine; a blue colour is produced. Solution.
C. To 5 ml of a saturated solution in water add 0.5 ml of ferric Chloroxylenol Solution contains not less than 4.75 per cent
chloride test solution; no blue colour is produced. and not more than 5.25 per cent of C8H9ClO.
D. Mix 50 mg with 0.5 g of anhydrous sodium carbonate and Tests
ignite strongly, cool, boil the residue with 5 ml of water, acidify
with nitric acid, filter and add 2 ml of silver nitrate solution; pH (2.4.24). 7.0 to 11.0.
a white precipitate is produced. Ethanol content (2.3.45). 16 to 21 per cent v/v.
300
IP 2007 CHLORPHENIRAMINE MALEATE
Assay. Determine by gas chromatography (2.4.13). A. Determine by infrared absorption spectrophotometry (2.4.6).
Test solution. Extract 4 ml of the solution under examination Compare the spectrum with that obtained with
with 20.0 ml of chloroform after adding 4 ml of 2 M chlorpheniramine maleate RS or with the reference spectrum
hydrochloric acid. Extract with two further quantities, each of chlorpheniramine maleate.
of 10.0 ml, of chloroform. Combine the chloroform extracts, B. When examined in the range 230 nm to 360 nm, a 0.002 per
shake with anhydrous sodium sulphate and filter. cent w/v solution in 0.05 M sulphuric acid shows an
absorption maximum only at about 265 nm; absorbance at
Reference solution (a). Dissolve 0.1 g of chloroxylenol RS in
about 265 nm, about 0.42 (2.4.7).
10.0 ml of a 0.8 per cent w/v solution of 4-chloro-o-cresol
(internal standard) in chloroform (solution A) and dilute to C. To 0.2 g add 3 ml of water and 1 ml of 10 M sodium hydroxide
20.0 ml with chloroform. and extract with three quantities, each of 5 ml, of ether. To
0.1 ml of the aqueous layer add a solution of 10 mg of
resorcinol in 3 ml of sulphuric acid and heat in a water-bath
solution but use 20.0 ml of solution A instead of 20 ml of
for 15 minutes; the solution is colourless. To the remainder of
chloroform.
the aqueous layer add 2 ml of bromine solution, heat in a
Chromatographic system water-bath for 15 minutes, heat to boiling and cool. To 0.2 ml
– a glass column 1.5m × 4 mm, packed with acid-washed, of the resulting solution add a solution of 10 mg of resorcinol
silanised diatomaceous support (80 to 100 mesh) coated in 3 ml of sulphuric acid and heat in a water-bath for 15 minutes;
with 3 per cent w/w of polyethylene glycol (such as a blue colour is produced.
Carbowax 20M),
– temperature: D. Dissolve 0.1 g in 10 ml of water and add dropwise with
column.160°, shaking 25 ml of a 1 per cent w/v solution of picric acid.
inlet port and detector. 220°, Collect the precipitate on a sintered-glass filter, wash with 3 ml
– a flame ionisation detector, of ethanol (95 per cent), recrystallise from ethanol (50 per
– flow rate. 30 ml per minute of the carrier gas. cent) and dry at 100° to 105°. The crystals melt between 196°
and 200° (2.4.21).
Calculate the content of C 8H 9ClO in the solution as a
percentage w/v. Tests
Labelling. The label states that the preparation is meant for Appearance of solution. A 10.0 per cent w/v solution is clear
external use only. (2.4.1), and not more intensely coloured than reference solution
BYS6 (2.4.1).
Chlorpheniramine Maleate Related substances. Determine by thin-layer chromatography
CH3
N Mobile phase. A mixture of 50 volumes of cyclohexane,
CH3 40 volumes of chloroform and 10 volumes of diethylamine.
N COOH
, Test solution. A 5 per cent w/v solution of the substance
Cl COOH under examination in chloroform.
C16H19ClN2,C4H4O4 Mol. Wt. 390.9 with chloroform and mix. Dilute 5 ml of the resulting solution
Chlorpheniramine Maleate is (RS)-3-(4-chlorophenyl)-3- to 25 ml with chloroform.
(pyrid-2-yl)propyldimethylamine hydrogen maleate. Apply to the plate 10 µl of each solution. After development,
Chlorpheniramine Maleate contains not less than 98.0 per cent dry the plate in air and examine in ultraviolet light at 254 nm.
and not more than 101.0 per cent of C16H19ClN2,C4H4O4, Any secondary spot in the chromatogram obtained with the
calculated on the dried basis. test solution is not more intense than the spot in the
chromatogram obtained with the reference solution. Ignore
Description. A white, crystalline powder; odourless.
any spot remaining on the line of application.
Identification Sulphated ash (2.3.18). Not more than 0.1 per cent.
Test A may be omitted if tests B, C and D are carried out. Tests Loss on drying (2.4.19). Not more than 0.5 per cent, determined
B, C and D may be omitted if test A is carried out. on 1.0 g by drying in an oven at 105° for 4 hours.
301
CHLORPHENIRAMINE INJECTION IP 2007
Assay. Weigh accurately about 0.2 g and dissolve in 20 ml of Related substances. Carry out the method described under
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric the Identification test using as the test solution a solution
acid, determining the end-point potentiometrically (2.4.25). prepared in the following manner. Evaporate an appropriate
Carry out a blank titration. volume of the injection to dryness in a current of nitrogen
using the minimum amount of heat. Dissolve the residue in
sufficient chloroform to produce a solution containing 5.0 per
C16H19ClN2,C4H4O4.
cent w/v of Chlorpheniramine Maleate and centrifuge. For the
Storage. Store protected from light and moisture. reference solution, dilute 1 volume of the test solution to
500 volumes with chloroform. After development, dry the plate
in air and spray with dilute potassium iodobismuthate
with the test solution is not more intense than the spot in the
Chlorpheniramine Injection chromatogram obtained with the reference solution.
Chlorpheniramine Maleate Injection Other tests. Complies with the tests stated under Parenteral
Chlorpheniramine Injection is a sterile solution of Preparations (Injections).
Chlorpheniramine Maleate in Water for Injections free from Assay. Dilute an accurately measured volume of the injection
dissolved air and containing suitable buffering and stabilising containing 10 mg of Chlorpheniramine Maleate to 500.0 ml
agents. with 0.25 M sulphuric acid. Measure the absorbance of the
Chlorpheniramine Injection contains not less than 90.0 per resulting solution at the maximum at about 265 nm (2.4.7).
cent and not more than 110.0 per cent of the stated amount of Calculate the content of C16H19ClN2,C4H4O4 taking 212 as the
chlorpheniramine maleate,C16H19ClN2,C4H4O4. specific absorbance at 265 nm.
Description. A colourless solution. Storage. Store protected from light.
Identification
plate with silica gel GF254. Heat the plate at 105° for 30
Chlorpheniramine Tablets
minutes before use. Chlorpheniramine Maleate Tablets
Mobile phase. A mixture of 50 volumes of ethyl acetate, Chlorpheniramine Tablets contain not less than 95.0 per cent
30 volumes of methanol and 20 volumes of 1 M acetic acid. and not more than 105.0 per cent of the stated amount of
chlorpheniramine maleate, C16H19ClN2,C4H4O4.
Test solution. Evaporate an appropriate volume of the injection
to dryness in a current of nitrogen using the minimum amount
Identification
of heat, dissolve the residue as completely as possible in
sufficient chloroform to produce a solution containing 0.5 per Determine by thin-layer chromatography (2.4.17), coating the
cent w/v of Chlorpheniramine Maleate and centrifuge. plate with silica gel GF254. Heat the plate at 105° for 30
minutes before use.
chlorpheniramine maleate RS in chloroform. Mobile phase. A mixture of 50 volumes of ethyl acetate,
30 volumes of methanol and 20 volumes of 1 M acetic acid.
dry the plate in air and examine in ultraviolet light at 254 nm. Test solution. Extract a quantity of the powdered tablets
The two principal spots in the chromatogram obtained with containing 5 mg of Chlorpheniramine Maleate with chloroform,
the test solution correspond to those in the chromatogram filter, evaporate the filtrate to dryness and dissolve the residue
obtained with the reference solution. Spray the plate with in 1 ml of chloroform.
dilute potassium iodobismuthate solution. The principal spot Reference solution. A 0.5 per cent w/v solution of
in the chromatogram obtained with the test solution chlorpheniramine maleate RS in chloroform.
reference solution. Apply to the plate 2 µl of each solution. After development,
Tests The two principal spots obtained in the chromatogram
obtained with the test solution correspond to those in the
pH (2.4.24). 4.0 to 5.2. chromatogram obtained with the reference solution. Spray
302
IP 2007 CHLORPROMAZINE HYDROCHLORIDE
the plate with dilute potassium iodobismuthate solution. The Chlorpromazine Hydrochloride
principal spot in the chromatogram obtained with the test
CH3
N
CH3
Tests
N Cl
Related substances. Determine by thin-layer chromatography ,HCl
(2.4.17), coating the plate with silica gel GF254. S
40 volumes of chloroform and 10 volumes of diethylamine. C17H19ClN2S,HCl Mol. Wt. 355.3
Chlorpromazine Hydrochloride is 2-chloro-10-(3-
Test solution. Extract a quantity of the powdered tablets
dimethylaminopropyl)phenothiazine hydrochloride.
containing 100 mg of Chlorpheniramine Maleate with
chloroform, filter, evaporate to dryness and dissolve the Chlorpromazine Hydrochloride contains not less than
residue in 2 ml of chloroform. 99.0 per cent and not more than 101.0 per cent of
C17H19ClN2S,HCl, calculated on the dried basis.
with chloroform and dilute 1.0 ml of the resulting solution to Description. A white or creamy-white, crystalline powder;
10.0 ml with the same solvent. odourless. It decomposes on exposure to air and light
becoming yellow, pink and finally violet.
Identification
test solution is not more intense than the spot in the Test A may be omitted if tests B, C and D are carried out. Test
chromatogram obtained with the reference solution. Ignore B may be omitted if tests A, C and D are carried out.
any spot remaining on the line of application.
Uniformity of content. Comply with test stated under Tablets. Compare the spectrum with that obtained with chlorpromazine
hydrochloride RS or with the reference spectrum of
Powder one tablet and carry out the Assay beginning at the
chlorpromazine hydrochloride.
words “shake with 20 ml of 0.05 M sulphuric acid....”. Calculate
the content of C16H19ClN2,C4H4O4 in the tablet. B. When examined in the range 230 nm to 360 nm, a 0.0005 per
cent w/v solution in 0.1 M hydrochloric acid shows
absorption maxima at about 254 nm and 306 nm; absorbance
Assay. Weigh and powder 20 tablets. Weigh accurately a at about 254 nm, 0.45 to 0.48 (2.4.7).
quantity of the powder containing about 4 mg of
C. Complies with the test for identification of phenothiazines
Chlorpheniramine Maleate, shake with 20 ml of 0.05 M
(2.3.3)
sulphuric acid for 5 minutes, add 20 ml of ether, shake carefully
and filter the acid layer into a second separator. Extract the D. A 5 per cent w/v solution gives reaction B of chlorides
ether layer with two quantities, each of 10 ml, of 0.05 M (2.3.1).
sulphuric acid, filter each acid layer into the second separator
and wash the filter with 0.05 M sulphuric acid. Make the Tests
combined acid extracts and washing just alkaline to litmus
pH (2.4.24). 3.5 to 4.5, determined in a 10.0 per cent solution.
paper with 1 M sodium hydroxide, add 2 ml in excess, and
extract with two quantities, each of 50 ml, of ether. Wash each Related substances (2.3.5). Use mobile phase (a).
ether extract with the same 20 ml of water and extract in Heavy metals (2.3.13). 2.0 g complies with the limit test for
succession with 20, 20 and 5 ml of 0.25 M sulphuric acid, heavy metals, Method B (10 ppm).
dilute the combined acid extracts to 50.0 ml with 0.25 M
sulphuric acid; dilute 10.0 ml to 50.0 ml with 0.25 M sulphuric Sulphated ash (2.3.18). Not more than 0.1 per cent.
acid and measure the absorbance of the resulting solution at Loss on drying (2.4.19). Not more than 0.5 per cent, determined
the maximum at about 265 nm (2.4.7). Calculate the content of on 1.0 g by drying in an oven at 105°.
C16H19ClN2,C4H4O4. taking 212 as the specific absorbance at
265 nm.
acetone and add 15 ml of mercuric acetate solution. Titrate
Storage. Store protected from light and moisture. with 0.1 M perchloric acid, using a saturated solution of
303
CHLORPROMAZINE INJECTION IP 2007
methyl orange in acetone as indicator. Carry out a blank Assay. Dilute an accurately measured volume of the injection
titration. with sufficient 0.1 M hydrochloric acid to produce a solution
1 ml of 0.1 M perchloric acid is equivalent to 0.03553 g of containing 0.0005 per cent w/v of Chlorpromazine
C17H19ClN2S,HCl. Hydrochloride and measure the absorbance of the resulting
Storage. Store protected from light and moisture. content of C17H19ClN2S,HCl, taking 915 as the specific
Chlorpromazine Injection
Chlorpromazine Hydrochloride Injection
Chlorpromazine Tablets
Chlorpromazine Injection is a sterile solution of Chlorpromazine
hydrochloride in Water for Injections free from air and Chlorpromazine Hydrochloride Tablets
containing buffering and stabilizing agents. Chlorpromazine Tablets contain not less than 92.5 per cent
Chlorpromazine Hydrochloride contains not less than and not more than 107.5 per cent of the stated amount of
95.0 per cent and not more than 105.0 per cent of the stated chlorpromazine hydrochloride, C17H19ClN2S,HCl. The tablets
amount of chlorpromazine hydrochloride, C17H19ClN2S,HCl. are coated.
NOTE — Protect the solutions from light throughout the NOTE — Protect the solutions from light throughout the
tests. tests.
Description. A colourless or almost colourless solution. Identification
Identification A. To a quantity of the powdered tablets containing 40 mg of
Chlorpromazine Hydrochloride add 10 ml of water and 2 ml of
A. To a volume containing 0.1 g of Chlorpromazine 10 M sodium hydroxide. Extract with 15 ml of ether and wash
Hydrochloride add 20 ml of water and 2 ml of 10 M sodium the ether extract with two quantities. each of 5 ml, of water, dry
hydroxide. Extract with 25 ml of ether, wash the ether extract with anhydrous sodium sulphate. evaporate the ether and
with two quantities, each of 5 ml, of water, dry the ether extract dissolve the residue in 0.4 ml of chloroform. The resulting
with anhydrous sodium sulphate, evaporate the ether and solution complies with the following test.
dissolve the residue in 1 ml of chloroform. The resulting
solution complies with the following test. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with chlorpromazine
Determine by infrared absorption spectrophotometry (2.4.6). hydrochloride RS treated in the same manner or with the
Compare the spectrum with that obtained with chlorpromazine reference spectrum of chlorpromazine hydrochloride.
reference spectrum of chlorpromazine hydrochloride. B. Digest a quantity of the powdered tablets containing 25 mg
of Chlorpromazine Hydrochloride with 25 ml of water and
B. Dilute a volume of the injection with sufficient 0.1 M
filter. Reserve a portion of the filtrate for Identification C. Dilute
hydrochloric acid to produce a solution containing
a volume of the filtrate with sufficient 0.1 M hydrochloric
0.0005 per cent w/v of Chlorpromazine Hydrochloride. The
acid to produce a solution containing 0.0005 per cent w/v of
resulting solution, when examined in the range 230 nm to
Chlorpromazine Hydrochloride. The resulting solution, when
360 nm shows absorption maxima at about 254 nm and 306 nm;
examined in the range 230 nm to 360 nm shows absorption
absorbance at about 254 nm, 0.45 to 0.48 (2.4.7).
maxima at about 254 nm and 306 nm; absorbance at about
C. Gives reaction B of chlorides (2.3.1). 254 nm, 0.45 to 0.48 (2.4.7).
Tests C. The filtrate reserved in test B gives reaction B of chlorides
(2.3.1).
Related substances (2.3.5). Use mobile phase (a).
Tests
Test solution. Dilute a volume of the injection with sufficient
of a mixture of 95 volumes of methanol and 5 volumes of Related substances. (2.3.5). Use mobile phase (a).
diethylamine to produce a solution containing 2.0 per cent of
Chlorpromazine Hydrochloride.
containing 0.2 g of Chlorpromazine Hydrochloride with 10 ml
Other tests. Complies with the tests stated under Parenteral of a mixture of 95 volumes of melhanol and 5 volumes of
Preparations (Injections). dielhylamine and filter.
304
IP 2007 CHLORPROPAMIDE
Reference solution. Dilute 1 volume of the test solution to A. Determine by infrared absorption spectrophotometry (2.4.6).
200 volumes with the same solvent mixture. Compare the spectrum with that obtained with chlorpropamide
Uniformity of content. Comply with the test stated under RS or with the reference spectrum of chlorpropamide.
Tablets. B. Dissolve 0.16 g in 50 ml of methanol, dilute 5 ml to 100 ml
Powder one tablet, shake with 1 ml of dilute hydrochloric and with 0.01 M hydrochloric acid and dilute 5 ml of this solution
40 ml of water for 15 minutes, add sufficient water to produce to 100 ml with 0.01 M hydrochloric acid. When examined in
100.0 m1 and mix. Centrifuge about 15 ml and to 10.0 ml of the the range 220 nm to 360 nm, the resulting solution shows an
clear, supernatant liquid add 2 ml of 1 M hydrochloric acid absorption maximum only at about 232 nm; absorbance at
and sufficient water to produce a solution containing about about 232 nm, about 0.48 (2.4.7).
0.0005 per cent w/v of Chlorpromazine Hydrochloride. Measure C. Boil 0.1 g with 8 ml of a 50 per cent w/w solution of sulphuric
the absorbance of the resulting solution at the maximum at acid under a reflux condenser for 30 minutes, cool and filter,
about 254 nm (2.4.7). Calculate the content of reserving the filtrate for test D. The precipitate, after
CI7H19ClN2S, HCl in the tablet taking 915 as the specific recrystallisation from water and drying, melts at about 143°
absorbance at 254 nm. (2.4.21).
Other tests. Comply with the tests stated under Tablets. D. Make the filtrate reserved in test C alkaline with sodium
Assay. Weigh and powder 20 tablets. Weigh accurately a hydroxide solution and heat; an ammoniacal odour is
quantity of the powder containing about 0.1 g of produced.
Chlorpromazine Hydrochloride, add 5 ml of dilute E. Heat 0.1 g with 1 g of anhydrous sodium carbonate at a dull
hydrochloric acid and 200 ml of water. Shake for 15 minutes red heat for 10 minutes. Cool, extract the residue with water
and add sufficient water to produce 500.0 mI. Centrifuge about and filter. Acidify the filtrate with dilute nitric acid and add
15 ml and to 5.0 ml of the clear, supernatant liquid add 10 ml of silver nitrate solution; a white precipitate is produced.
dilute hydrochloric acid and sufficient water to produce
200.0 ml. Measure the absorbance of the resulting solution at Tests
the maximum at about 254 nm (2.4.7). Calculate the content of
C17HI9CIN2S,HCI, taking 915 as the specific absorbance at Related substances. Determine by thin-layer chromatography
254 nm. (2.4.17), coating the plate with silica gel G.
Storage. Store protected from light. Mobile phase. A mixture of 100 volumes of chloroform,
50 volumes of methanol, 30 volumes of cyclohexane and
11.5 volumes of strong ammonia solution.
Chlorpropamide examination in 10 ml of acetone.
O O O
4-chlorobenzenesulphonamide in acetone.
S CH3
N N Reference solution (b). A 0.02 per cent w/v solution of
H H 1,3-dipropylurea RS in acetone.
Cl
C10H13ClN2O3S Mol. Wt. 276.7 substance under examination in acetone.
Chlorpropamide is 1-(4-chlorobenzenesulphonyl)-3- Apply to the plate 5 µl of each solution. After development,
propylurea. dry the plate in a current of cold air, heat at 110° for 10 minutes,
Chlorpropamide contains not less than 99.0 per cent and not place the plate, while hot, in a tank of chlorine gas prepared by
more than 101.0 per cent of C10H13ClN2O3S, calculated on the adding hydrochloric acid to a 5 per cent w/v solution of
dried basis. potassium permanganate contained in a beaker placed in the
tank and allow to stand for 2 minutes. Dry it in a current of
Description. A white, crystalline powder; odourless or almost cold air until an area of the plate below the line of application
odourless. gives at most a very faint blue colour with a 0.5 per cent w/v
solution of potassium iodide in starch solution; avoid
Identification
prolonged exposure to cold air. Any spots corresponding to
Test A may be omitted if tests B, C, D and E are carried out. 4-chlorobenzenesulphonamide and 1,3-dipropylurea in the
Tests B, C, D and E may be omitted if test A is carried out. chromatogram obtained with the test solution are not more
305
CHLORPROPAMIDE TABLETS IP 2007
intense than the spots in the chromatogram obtained with Test solution. Shake a quantity of the powdered tablets
reference solutions (a) and (b) respectively. Any other containing 0.6 g of Chlorpropamide with 10 ml of acetone and
secondary spot in the chromatogram obtained with the test filter.
solution is not more intense than the spot in the chromatogram Reference solution (a). A 0.02 per cent w/v solution of
obtained with reference solution (c). 4-chlorobenzenesulphonamide in acetone.
Heavy metals (2.3.13). 0.66 g complies with the limit test for Reference solution (b). A 0.02 per cent w/v solution of
heavy metals, Method B (30 ppm). 1,3-dipropylurea RS in acetone.
Sulphated ash (2.3.18). Not more than 0.1 per cent, determined Reference solution (c). A 0.02 per cent w/v solution of the
on 2.0 g. substance under examination in acetone.
Loss on drying (2.4.19). Not more than 1.5 per cent, determined Apply to the plate 5 µl of each solution. After development,
on 1.0 g by drying in an oven at 105°. dry the plate in a current of cold air, heat at 110° for 10 minutes,
Assay. Weigh accurately about 0.5 g and dissolve in 50 ml of place the plate, while hot, in a tank of chlorine gas prepared by
ethanol (95 per cent) previously neutralised to adding hydrochloric acid to a 5 per cent w/v solution of
phenolphthalein solution. Add 25 ml of water and titrate with potassium permanganate contained in a beaker placed in the
0.1 M sodium hydroxide using phenolphthalein solution as tank and allow to stand for 2 minutes. Dry it in a current of
indicator. cold air until an area of the plate below the line of application
gives at most a very faint blue colour with a 0.5 per cent w/v
solution of potassium iodide in starch solution; avoid
C10H13ClN2O3S.
prolonged exposure to cold air. Any spots corresponding to
4-chlorobenzenesulphonamide and 1,3-dipropylurea in the
chromatogram obtained with the test solution are not more
Chlorpropamide Tablets intense than the spots in the chromatogram obtained with
Chlorpropamide Tablets contain not less than 92.5 per cent reference solutions (a) and (b) respectively. Any other
and not more than 107.5 per cent of the stated amount of secondary spot in the chromatogram obtained with the test
chlorpropamide, C10H13ClN2O3S. solution is not more intense than the spot in the chromatogram
obtained with reference solution (c).
Identification Dissolution (2.5.2).
Extract a quantity of the powdered tablets containing 1 g of Apparatus. No 1
Chlorpropamide with five quantities, each of 4 ml, of acetone, Medium. 900 ml of a 0.68 per cent w/v solution of potassium
filter and carefully evaporate the filtrate to dryness on a water- dihydrogen phosphate adjusted to pH 7.4 by the addition of
bath. The residue complies with the following tests. 1 M sodium hydroxide
A. Boil 0.1 g with 8 ml of a 50 per cent w/w solution of sulphuric Speed and time. 100 rpm and 60 minutes.
acid under a reflux condenser for 30 minutes, cool and filter, Withdraw a suitable volume of the medium and filter through
reserving the filtrate for test B. The precipitate, after a membrane filter with an average pore diameter not greater
recrystallisation from water and drying, melts at about 143° than 1.0 µm. Reject the first few ml of the filtrate and dilute a
(2.4.21). suitable volume of the filtrate with 0.1 M hydrochloric acid to
B. Make the filtrate reserved in test A alkaline with sodium obtain a solution containing about 10 µg of chlorpropamide
hydroxide solution and heat; an ammonical odour is produced. per ml. Measure the absorbance of the resulting solution at
C. Heat 0.1 g with 1 g of anhydrous sodium carbonate at a dull
C10H13ClN2O3S taking 598 as the specific absorbance at
red heat for 10 minutes. Cool, extract the residue with water
232 nm.
and filter. Acidify the filtrate with dilute nitric acid and add
silver nitrate solution; a white precipitate is produced. D. Not less than 75 per cent of the stated amount of
C10H13ClN2O3S.
Tests
Mobile phase. A mixture of 100 volumes of chloroform, Chlorpropamide and shake with 40 ml of methanol for
50 volumes of methanol, 30 volumes of cyclohexane and 20 minutes, add sufficient methanol to produce 50.0 ml, mix,
11.5 volumes of strong ammonia solution. filter and dilute 5.0 ml of the filtrate to 100.0 ml with 0.1 M
306
IP 2007 CHLORTHALIDONE
hydrochloric acid. Mix, dilute 10.0 ml of this solution to 250.0 D. Dissolve 10 mg in 1 ml of sulphuric acid; an intense yellow
ml with 0.1 M hydrochloric acid and measure the absorbance colour is produced.
of the resulting solution at the maximum at about 232 nm (2.4.7).
Calculate the content of C10H13ClN2O3S taking 598 as the Tests
specific absorbance at 232 nm. Appearance of solution. Dissolve 1.0 g in sufficient 2 M sodium
hydroxide to produce 10 ml. The solution is clear (2.4.1), and
not more intensely coloured than degree 6 of the appropriate
range of reference solutions (2.4.1).
Chlorthalidone
Acidity. Dissolve 1 g in a mixture of 25 ml of acetone and 25 ml
O of carbon dioxide-free water with the aid of heat, cool and
titrate with 0.01 M sodium hydroxide using methyl red solution
NH O O as indicator. Repeat the operation without the substance under
S examination. The difference between the titrations is not more
HO NH2 than 0.75 ml.
Cl
C14H11ClN2O4S Mol. Wt 338.8 Mobile phase. A mixture of 30 volumes of dioxan, 30 volumes
Chlorthalidone is (RS)-2-chloro-5-(1-hydroxy-3- of 2-propanol, 30 volumes of toluene and 20 volumes of strong
oxoisoindolin-3-yl)benzenesulphonamide. ammonia solution.
Chlorthalidone contains not less than 98.0 per cent and not Test solution. A 2 per cent w/v solution of the substance
more than 102.0 per cent of C14H11ClN2O4S, calculated on the under examination in acetone.
dried basis. Reference solution (a). Dilute 10 ml of the test solution to
Description. A white to yellowish-white, crystalline powder; 20 ml with acetone and mix. Dilute 1 ml of the resulting solution
almost odourless. to 100 ml with acetone.
Reference solution (b). A 0.02 per cent w/v solution of 2-(4-
Identification chloro-3-sulphamoylbenzoyl)benzoic acid RS in acetone.
Test A may be omitted if tests B, C and D are carried out. Tests Apply to the plate 10 µl of each solution. After development,
B, C and D may be omitted if test A is carried out. dry the plate in air and examine in ultraviolet light at 254 nm.
Any spot corresponding to 2-(4-chloro-3-sulphamoyl-
A. Determine by infrared absorption spectrophotometry (2.4.6). benzoyl)benzoic acid in the chromatogram obtained with the
Compare the spectrum with that obtained with chlorthalidone test solution is not more intense than the spot in the
RS or with the reference spectrum of chlorthalidone. chromatogram obtained with reference solution (b) and any
B. When examined in the range 230 nm to 360 nm, a 0.01 per other secondary spot is not more intense than the spot in the
cent w/v solution in ethanol (95 per cent) shows absorption chromatogram obtained with reference solution (a).
maxima at about 275 nm and at about 284 nm; absorbance at Heavy metals (2.3.13). 2.0 g complies with the limit test for
about 275 nm, about 0.6 and at about 284 nm, about 0.45 (2.4.7). heavy metals, Method B (10 ppm).
C. Determine by thin-layer chromatography (2.4.17), coating Chlorides (2.3.12). Triturate 0.5 g with 30 ml of water, shake
the plate with silica gel GF254. for 5 minutes and filter. 15 ml of the filtrate complies with the
Mobile phase. A mixture of 197 volumes of ethyl acetate and limit test for chlorides. Use 5.0 ml of chloride standard solution
3 volumes of water. (25 ppm Cl) to prepare the standard (500 ppm).
Test solution. Dissolve 0.1 g of the substance under Sulphated ash (2.3.18). Not more than 0.1 per cent.
examination in 100 ml of acetone. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Reference solution. A 0.1 per cent w/v solution of on 1.0 g by drying in an oven at 105°.
chlorthalidone RS in acetone. Assay. Weigh accurately about 0.2 g and dissolve in 50 ml of
acetone. Titrate with 0.1 M tetrabutylammonium hydroxide
in an atmosphere of nitrogen, determining the end-point
solution corresponds to that in the chromatogram obtained 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
with the reference solution. 0.03388 g of C14H11ClN2O4S.
307
CHLORTHALIDONE TABLETS IP 2007
Chlorthalidone Tablets 275 nm (2.4.7). Calculate the content of C14H11ClN2O4S taking

57.4 as the specific absorbance at 275 nm.
Chlorthalidone Tablets contain not less than 92.5 per cent and
chlorthalidone, C14H11ClN2O4S.
Identification
Cholecalciferol
Vitamin D3
Heat a quantity of the powdered tablets containing 0.2 g of
Chlorthalidone with 20 ml of acetone on a water-bath for H3C CH3
10 minutes, cool and filter. Add 40 ml of water to the filtrate H3C
and heat on a water-bath for 20 minutes using a gentle current CH3
of air to remove the solvent. Cool to room temperature and H2C
allow to stand, filter and dry the crystals at 105° for 4 hours.
The crystals comply with the following tests. H
A. Determine by infrared absorption spectrophotometry (2.4.6). HO
Compare the spectrum with that obtained with chlorthalidone
RS or with the reference spectrum of chlorthalidone. C27H44O Mol Wt. 384.6
B. When examined in the range 230 nm to 360 nm, a 0.01 per Cholecalciferol is (5Z,7E)-(3S)-9,10-secocholesta-5,7,10(19)-
cent w/v solution in ethanol (95 per cent) shows absorption trien-3-ol.
maxima at about 275 nm and at about 284 nm; absorbance at Cholecalciferol contains not less than 97.0 per cent and not
about 275 nm, about 0.6 and at about 284 nm, about 0.45 (2.4.7). more than 103.0 per cent of C27H44O.
C. Wash with water a quantity of the crystals obtained in test Description. White or almost white crystals; odourless or
A and dissolve 50 mg in 3 ml of sulphuric acid; an intense almost odourless. It is sensitive to air, heat and light. A
yellow colour is produced. reversible isomerisation to precholecalciferol may occur in
solution, depending on temperature and time.
Tests
Identification
(2.4.17), coating the plate with silica gel GF254. Test A may be omitted if tests B, C and D are carried out. Tests
containing 50 mg of Chlorthalidone with 5 ml of acetone, A. Determine by infrared absorption spectrophotometry (2.4.6).
centrifuge and use the supernatant liquid. Compare the spectrum with that obtained with cholecalciferol
RS.
2-(4-chloro-3-sulphamoylbenzoyl)benzoic acid RS in B. Dissolve 1 mg in 1 ml of 1,2-dichloroethane and 4 ml of
acetone. antimony trichloride solution; a yellowish-orange colour is
produced.
dry the plate in air and examine in ultraviolet light at 254 nm. C. In the test for 7-Dehydrocholesterol, the principal spot in
Any secondary spot in the chromatogram obtained with the the chromatogram obtained with the test solution corresponds
test solution is not more intense than the spot in the to that in the chromatogram obtained with reference solution
chromatogram obtained with the reference solution. (b).
Other tests. Comply with the tests stated under Tablets. D. To a solution of about 0.5 mg in 5 ml of chloroform add
0.3 ml of acetic anhydride and 0.1 ml sulphuric acid and
shake vigorously; a bright red colour is produced which rapidly
changes through violet and blue to green.
Chlorthalidone, boil with 30 ml of methanol under a reflux
condenser for 5 minutes, shake vigorously for 15 minutes, Tests
cool and filter; wash the residue with methanol and filter.
Dilute the combined filtrate and washings to 100.0 ml with Specific optical rotation (2.4.22). +105° to +112°, determined,
methanol. To 5.0 ml add 2 ml of 1 M hydrochloric acid and within 30 minutes of preparation, in a solution prepared by
sufficient methanol to produce 50.0 ml. Measure the dissolving 0.2 g rapidly and without heating in sufficient
absorbance of the resulting solution at the maximum at about aldehyde-free ethanol (95 per cent) to produce 25.0 ml.
308
IP 2007 CHORIONIC GONADOTROPHIN
Light absorption. Dissolve 10 mg, rapidly and without heating, Reference solution (b). Reflux 5.0 ml of solution A, under
in sufficient aldehyde-free ethanol (95 per cent) to produce nitrogen, on a water-bath for 60 minutes to obtain a solution
100.0 ml. Dilute 5.0 ml of this solution to 50.0 ml with aldehyde- of cholecalciferol, precholecalciferol and trans-cholecalciferol.
free ethanol (95 per cent). Absorbance of the resulting Chromatographic system
solution at the maximum at about 265 nm, measured within – a stainless steel column 25 cm × 4.6 mm, packed with
30 minutes of preparation, 0.46 to 0.50 (2.4.7). porous silica particles (5 µm) (such as Nucleosil 50-S
7-Dehydrocholesterol. Determine by thin-layer 5 µm),
chromatography (2.4.17), coating the plate with silica gel G. – mobile phase: a mixture of 997 volumes of hexane and
Mobile phase. A 0.01 per cent w/v solution of butylated 3 volumes of 1-pentanol,
hydroxytoluene in a mixture of equal volumes of cyclohexane – flow rate. 2 ml per minute,
and peroxide-free ether. – spectrophotometer set at 254 nm,
Inject reference solution (b) and record the chromatogram
adjusting the sensitivity so that the height of the peak due to
examination in sufficient of 1,2-dichloroethane containing
cholecalciferol is more than 50 per cent of full-scale deflection.
1 per cent w/v of squalane and 0.01 per cent w/v of butylated
The approximate relative retention times calculated with
hydroxytoluene (solvent A) to produce 5 ml.
reference to cholecalciferol are 0.4 for precholecalciferol and
Reference solution (a). A solution containing 0.005 per cent 0.5 for trans-cholecalciferol. The resolution between
w/v of 7-dehydrocholesterol RS in solvent A. precholecalciferol and trans-cholecalciferol should be not less
Reference solution (b). A solution containing 2.5 per cent than 1.0; if necessary adjust the proportions of the
w/v of cholecalciferol RS in solvent A. constituents and flow rate of the mobile phase to obtain the
required resolution.
Reference solution (c). Mix equal volumes of reference
solutions (a) and (b). Inject reference solution (a) and record the chromatogram
Apply to the plate 10 µl of each solution. Develop the adjusting the sensitivity so that the height of the peak due to
chromatograms immediately, protected from light. After cholecalciferol is more than 50 per cent of full-scale deflection.
development, dry the plate in air and spray three times with Inject the test solution. Measure the areas for the major peaks.
antimony trichloride reagent. Examine the chromatograms
Calculate the content of C27H44O.
for not more than 4 minutes after spraying. The principal spot
in the chromatogram obtained with the test solution is initially Storage. Store protected from light in hermetically sealed
orange-yellow but becomes brown later. In the chromatogram containers under nitrogen in a refrigerator. The contents of an
obtained with the test solution any violet spot with an Rf opened container should be used immediately.
value slightly lower than that of the principal spot (due to 7-
dehydrocholesterol and appearing slowly) is not more intense
solution (a). The test is not valid unless the chromatogram Chorionic Gonadotrophin
obtained with reference solution (c) shows two clearly
Human Chorionic Gonadotrophin
separated principal spots.
Chorionic Gonadotrophin is a dry, sterile preparation of
Assay. Carry out the following procedure as rapidly as
placental glycoproteins that has luteinising activity. It is
possible in subdued light and protected from air.
extracted from the urine of pregnant women. The material is
Determine by liquid chromatography (2.4.14). sterilised by filtration and dried under reduced pressure or
Test solution. Weigh accurately about 50.0 mg of the substance freeze-dried.
under examination, dissolve in 10 ml of toluene without heating Chorionic Gonadotrophin contains not less than 2500 Units
and dilute to 100.0 ml with the mobile phase; dilute 5.0 ml of per mg.
this solution to 50.0 ml with the mobile phase; further dilute
5.0 ml of this solution to 50.0 ml with the mobile phase. Description. A white or almost white, amorphous powder.
Reference solution (a). Dissolve 50.0 mg of cholecalciferol Identification
RS in 10 ml of toluene without heating and dilute to 100.0 ml
with the mobile phase; dilute 5.0 ml of this solution to 50.0 ml It causes an increase in the weight of the seminal vesicles or
with the mobile phase (Solution A); further dilute 5.0 ml of of the prostate glands of immature male rats when administered
solution A to 50.0 ml with the mobile phase. as directed in the Assay.
309
CHORIONIC GONADOTROPHIN IP 2007
Tests antimicrobial preservative such as 0.4 per cent w/v of phenol

or 0.002 per cent w/v of thiomersal. Store the solution at a
Appearance of solution. A 1.0 per cent w/v solution is clear temperature of 2° to 8°.
Use immature male rats of the same strain, approximately
Water. Not more than 5 per cent, determined by the following 21 days old and of approximately equal weight within the
method. range 25 to 35 g. Assign the rats at random to four equal
Determine by gas chromatography (2.4.13). groups of at least eight animals. If sets of four littermates are
available, allot one littermate from each set at random to each
Use throughout dry glassware that may be siliconised. group and mark according to the litter.
Internal standard. Dilute 15 µl of anhydrous methanol with Choose two doses of the standard preparation and two of the
sufficient anhydrous 2-propanol to produce 100 ml. test solution such that the smaller dose is sufficient to produce
Test solution (a). Dissolve 4 mg of the substance under a positive response in some of the rats and the larger dose
examination in 0.5 ml of anhydrous 2-propanol. does not produce a maximum response in all of the rats. As an
initial approximation, doses of 7.5 and 15 Units may be tried
Test solution (b). Dissolve 4 mg of the substance under although the dose will depend on the sensitivity of the animals
examination in 0.5 ml of test solution (a). used, which may vary widely.
Reference solution. Add 10 µl of water to 50 ml of test solution Inject subcutaneously into each rat the daily dose allocated
(a). to its group on 4 consecutive days at the same time each day.
Chromatographic system On the fifth day, about 24 hours after the last injection, kill the
– a stainless steel column 1m × 2 mm, packed with porous rats and remove the seminal vesicles or the prostate glands
polymer beads (60 to 80 mesh) (such as Chromosorb from each animal. Remove any extraneous fluid and tissue
102), from the vesicles or glands and weigh them immediately.
– temperature: Calculate the result of the assay by standard statistical
column.120°, methods using the weight of the vesicles or prostate glands
inlet port and detector. 150°, as the response.
– thermal conductivity detector, The estimated potency is not less than 80 per cent and not
– flow rate. 30 ml per minute of the carrier gas (helium). more than 125 per cent of the stated potency. The fiducial
From the chromatograms obtained, and taking into account limits of error are not less than 64 per cent and not more than
any water detectable in test solution (a), calculate the 156 per cent of the stated potency.
percentage of water taking 0.9960 g as the weight per ml at 25°. Chorionic Gonadotrophin intended for use in the
Assay. Carry out the biological assay of chorionic manufacture of parenteral preparations without a further
gonadotrophin described below. appropriate procedure for the removal of bacterial
Standard preparation. The 3rd International Standard for requirement.
Chorionic Gonadotrophin, human, established in 1986,
Bacterial endotoxins (2.2.3). Not more than 15 Endotoxin Units
consisting of a freeze-dried extract of human chorionic
per ml of a solution prepared in the following manner. Dissolve
gonadotrophin with human albumin (supplied in ampoules
a quantity in water BET to obtain a solution containing
containing 650 Units), or another suitable preparation the
500 units of chorionic gonadotrophin per ml. Carry out the
potency of which has been determined in relation to the
test using Maximum Valid dilution of this solution calculated
International Standard.
from the declared sensitivity of the lysate used in the test.
Dissolve a sufficient quantity corresponding to the daily doses
Chorionic Gonadotrophin intended for use in the
to be used in sufficient albumin-phosphate buffer pH 7.2 so
that the daily dose is about 0.2 ml. Add a suitable antimicrobial
appropriate sterilisation procedure complies with the
preservative such as 0.4 per cent w/v of phenol or 0.002 per
following additional requirements.
cent w/v of thiomersal. Store the solution at a temperature of
2° to 8°. Sterility (2.2.11). Complies with the test for sterility.
Test preparation. Dissolve a sufficient quantity of the Abnormal toxicity (2.2.1). Complies with the test for abnormal
preparation under examination corresponding to the daily toxicity (2.2.1) using a quantity equivalent to 1000 Units
doses to be used in sufficient albumin-phosphate buffer dissolved in 0.5 ml of sodium chloride injection and observing
pH 7.2 so that the daily dose is about 0.2 ml. Add a suitable the animals for 48 hours.
310
IP 2007 CHORIONIC GONADOTROPHIN INJECTION
Storage. Store protected from light in a tamper-evident Test solution (a). Dissolve 4 mg of the substance under
container, which is sealed so as to exclude micro-organisms, examination in 0.5 ml of anhydrous 2-propanol.
in a refrigerator (2° to 8°). Test solution (b). Dissolve 4 mg of the substance under
Labelling. The label states (1) the number of Units contained examination in 0.5 ml of test solution (a).
in the container; (2) the number of Units per mg; (3) whether Reference solution. Add 10 µl of water to 50 ml of test solution
or not it is intended for use in the manufacture of parenteral (a).
preparations.
– a stainless steel column 1m x 2 mm, packed with porous
polymer beads (60 to 80 mesh) (such as Chromosorb
102),
Chorionic Gonadotrophin Injection – temperature:
Chorionic Gonadotrophin Injection is a sterile material column.120°,
consisting of Chorionic Gonadotrophin with or without inlet port and detector. 150°,
excipients such as buffers, diluents or other inert substances – thermal conductivity detector,
such as Lactose or Sodium Chloride. It may also contain an – flow rate. 30 ml per minute of the carrier gas (helium).
antimicrobial agent. It is filled in a sealed container. From the chromatograms obtained, and taking into account
The injection is constituted by dissolving the contents of the any water detectable in test solution (a), calculate the
sealed container in the requisite amount of sterile Water for percentage of water taking 0.9960 g as the weight per ml at 25°.
Injections, immediately before use. Assay. Carry out the biological assay of chorionic
The constituted solution complies with the requirements for gonadotrophin described below.
Clarity of solution and Particulate matter stated under Standard preparation. The 3rd International Standard for
Parenteral Preparations (Injections). Chorionic Gonadotrophin, human, established in 1986,
Storage. The constituted solution should be used immediately consisting of a freeze-dried extract of human chorionic
after preparation but, in any case, within the period gonadotrophin with human albumin (supplied in ampoules
recommended by the manufacturer. containing 650 Units), or another suitable preparation the
potency of which has been determined in relation to the
Chorionic Gonadotrophin Injection contains not less than International Standard.
Dissolve a sufficient quantity corresponding to the daily doses
potency.
to be used in sufficient albumin-phosphate buffer pH 7.2 so
Description. A white or almost white powder. that the daily dose is about 0.2 ml. Add a suitable antimicrobial
The contents of the sealed container comply with the preservative such as 0.4 per cent w/v of phenol or 0.002 per
requirements for Powders for Injections stated under cent w/v of thiomersal. Store the solution at a temperature of
Parenteral Preparations and with the following 2° to 8°.
requirements. Test preparation. Dissolve a sufficient quantity of the injection
under examination corresponding to the daily doses to be
Identification used in sufficient albumin-phosphate buffer pH 7.2 so that
It causes an increase in the weight of the seminal vesicles or the daily dose is about 0.2 ml. Add a suitable antimicrobial
of the prostate glands of immature male rats when administered preservative such as 0.4 per cent w/v of phenol or 0.002 per
as directed in the Assay. cent w/v of thiomersal. Store the solution at a temperature of
2° to 8°.
Tests Use immature male rats of the same strain, approximately
21 days old and of approximately equal weight within the
pH (2.4.24). 6.0 to 8.0, determined in a 1.0 per cent w/v solution. range 25 to 35 g. Assign the rats at random to four equal
Water. Not more than 5.0 per cent, determined by the following groups of at least eight animals. If sets of four littermates are
method. available, allot one littermate from each set at random to each
group and mark according to the litter.
Choose two doses of the standard preparation and two of the
Use throughout dry glassware that may be siliconised.
test solution such that the smaller dose is sufficient to produce
Internal standard. Dilute 15 µl of anhydrous methanol with a positive response in some of the rats and the larger dose
sufficient anhydrous 2-propanol to produce 100 ml. does not produce a maximum response in all of the rats. As an
311
CICLESONIDE IP 2007
initial approximation, doses of 7.5 and 15 Units may be tried Ciclesonide contains not less than 98.0 per cent and not more
although the dose will depend on the sensitivity of the animals than 102.0 per cent of ciclesonide, C32H44O7, calculated on the
used that may vary widely. anhydrous basis.
Inject subcutaneously into each rat the daily dose allocated Description. A white to-off white powder.
to its group on 4 consecutive days at the same time each day.
On the fifth day, about 24 hours after the last injection, kill the Identification
rats and remove the seminal vesicles or the prostate glands
from each animal. Remove any extraneous fluid and tissue A. Determine by infrared absorption spectrophotometry (2.4.6).
from the vesicles or glands and weigh them immediately. Compare the spectrum with that obtained with ciclesonide RS
Calculate the result of the assay by standard statistical or with the reference spectrum of ciclesonide.
methods using the weight of the vesicles or prostate glands B. In the Assay, the principal peak in the chromatogram
as the response. obtained with the test solution corresponds to the principal
The estimated potency is not less than 80 per cent and not peak in the chromatogram obtained with the reference solution.
more than 125 per cent of the stated potency. The fiducial
limits of error are not less than 64 per cent and not more than Tests
156 per cent of the stated potency.
Specific optical rotation (2.4.22). +90.0º to +98.0º, determined
Bacterial endotoxins (2.2.3). Not more than 15 Endotoxin Units in a 0.5 per cent w/v solution in methanol.
per ml of a solution prepared in the following manner. Dissolve
the contents of a sealed container in water BET to obtain a Related substances. Determine by liquid chromatography
solution containing 500 units of chorionic gonadotrophin per (2.4.14).
ml. Carry out the test using Maximum Valid Dilution of this Test solution. Dissolve 50 mg of the substance under
solution calculated from the declared sensitivity of the lysate examination in 50 ml of methanol.
used in the test.
Abnormal toxicity (2.2.1). Use a quantity equivalent to ciclesonide RS in methanol.
1000 Units dissolved in 0.5 ml of sodium chloride injection
and observing the animals for 48 hours. Chromatographic system
Storage. Store protected from light in containers, which are octylsilyl silica gel (5 µm),
sealed so as to exclude micro-organisms, at a temperature not – mobile phase: A. dilute 1 volume of orthophosphoric
exceeding 20°. acid to 1000 ml with water,
Labelling. The label states (1) the number of Units contained B. acetonitrile
in the sealed container; (2) the name(s) of any added – flow rate. 1.5 ml per minute,
substance(s). – a linear gradient programme using the conditions given
below,
Ciclesonide – a 20 µl loop injector.
O (min.) (per cent v/v) (per cent v/v)
CH3 0 65 35
O 20 25 75
H3C H CH
3
HO O 40 25 75
H3C H O 45 65 35
55 65 35
H H
O inject reference solution. The test is not valid unless the tailing
factor is not more than 2.0 and the column efficiency in not
C32H44O7 Mol. Wt. 540.7 less than 30000 theoretical plates.
Ciclesonide is (11β,16α)-16,17-[(R)-cyclohexylmethylene] Inject the test solution. Any individual impurity is not more
bis(oxy)-11-hydroxy-21-(2-methyl-1-oxopropoxy)pregna-1,4- than 0.5 per cent and the sum of all the impurities found is not
diene-3,20-dione. more than 1.0 per cent.
312
IP 2007 CICLESONIDE INHALATION
Heavy metals (2.3.13). 1.0 g complies with the limit test for Tests
Other tests. Complies with the tests stated under Inhalation
Sulphated ash (2.3.18). Not more than 0.1 per cent, determined Preparations (Pressurised Metered-dose Preparations).
on 1.0 g.
Follow the procedure described under Assay with suitable
Water (2.3.43). Not more than 0.5 per cent, determined on dilution of the reference solution wherever the amount of
1 g. active substance is to be determined in any test.
Assay. Determine by liquid chromatography (2.4.14). Assay. Carry out the test for Content of active ingredient
delivered per actuation stated under Inhalation Preparations
(Pressurised Metered-dose Preparations).
examination in 50.0 ml of methanol. Dilute 5 ml of the resulting
solution to 50.0 ml with methanol. Determine by liquid chromatography (2.4.14).
Reference solution. A 0.01 per cent w/v solution of ciclesonide Solvent mixture. A mixture of equal volumes of water and
RS in methanol. acetonitrile.
Chromatographic system Test solution. Prepare using the solvent mixture as described
– a stainless steel column 25 cm x 4.6 mm, packed with under the test for Content of active ingredient delivered per
octylsilyl silica gel (5 µm), actuation stated under Inhalation Preparations (Pressurised
– mobile phase: a mixture of 30 volumes of 0.1 per cent Metered-dose Preparations).
orthophosphoric acid and 70 volumes of Reference solution (a). A 0.04 per cent w/v solution of
acetonitrile, ciclesonide RS in acetonitrile.
– flow rate. 1.5 ml per minute, Reference solution (b). Dilute reference solution (a) with the
– spectrophotometer set at 245 nm, solvent mixture to obtain a solution containing 32 µg of
– a 20 µl loop injector. Ciclesonide per ml.
Inject the reference solution. The test is not valid unless relative Chromatographic system
standard deviation for replicate injections is not more than 1.0 – a stainless steel column 15 cm x 4.6 mm, packed with
per cent. octylsilyl silica gel (5 µm),
Inject the test solution and the reference solution. – mobile phase: a mixture of 30 volumes of a buffer solution
prepared by diluting 1 ml of orthophosphoric acid to
Calculate the content of C32H44O7. 1000 ml with water, and 70 volumes of acetonitrile,
Storage. Store protected from light, at a temperature not – flow rate. 3 ml per minute,
exceeding 30º. – spectrophotometer set at 245 nm,
– inject 200 µl.
the tailing factor is not more than 2.0 and the relative standard
Ciclesonide Inhalation deviation for replicate injections is not more than 2.0 per cent.
Ciclesonide Inhalation is a suspension of microfine Ciclesonide Inject the test solution and reference solution (b).
in a suitable liquid filled in a suitable pressurized container. It Calculate the content of C32H44O7 in the solution and the
may contain suitable pharmaceutical aids such as surfactants, amount of C32H44O7 delivered per actuation of the valve.
stabilizing agents.
Determine the content of active ingredient a second and third
Ciclesonide Inhalation delivers not less than 80.0 per cent and time by repeating the procedure on the middle ten and on the
not more than 120.0 per cent of the stated amount of last ten successive combined actuations of the valve. For
ciclesonide, C32H44O7, per inhalation by actuation of the each of the three determinations the average content of
valve. C32H44O7 delivered per actuation of the valve meets the
requirements.
Identification Storage. Store protected from moisture at a temperature not
In the Assay the principal peak in the chromatogram obtained exceeding 30º.
with the test solution corresponds to the principal peak in the Labelling. The label states the amount of active ingredient
chromatogram obtained with reference solution (b). delivered per inhalation.
313
CIMETIDINE IP 2007
Cimetidine Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with

methanol.
H Reference solution (a). Dilute 1 ml of solution (a) to 100 ml
N N NHCH3 with methanol and dilute 20 ml of this solution to 100 ml with
S
methanol.
HN NCN
CH3 Reference solution (b). Dilute 5 ml of reference solution (a) to
C10H16N6S. Mol Wt. 252.3 Reference solution (c). Dilute 5 ml of reference solution (b) to
Cimetidine is 2-cyano-1-methyl-3-[2-(5-methylimidazol-4- 10 ml with methanol.
ylmethylthio)ethyl]guanidine. Reference solution (d). Dissolve 10 mg of cimetidine RS in
Cimetidine contains not less than 98.5 per cent and not more 2 ml of methanol.
than 101.5 per cent of C10H16N6S, calculated on the dried basis. Apply separately to two plates 4 µl of each solution. Allow
Description. A white or almost white powder. the first plate to stand for 15 minutes in the tank saturated
with vapour from mobile phase (a). Develop the second plate
Identification using mobile phase (b). After development, dry the plates in a
current of air, expose to iodine vapour until maximum contrast
of the spots has been obtained and examine in ultraviolet light
at 254 nm. The following limits apply to both methods. Any
A. Determine by infrared absorption spectrophotometry (2.4.6), secondary spot in the chromatogram obtained with test
using a potassium bromide dispersion obtained from the solid solution (a) is not more intense than the spot in the
state without prior solvent treatment. Compare the spectrum chromatogram obtained with reference solution (a) and not
with that obtained with cimetidine RS or with the reference more than two such spots are more intense than the spot in
spectrum of cimetidine. No shoulder or peak should be the chromatogram obtained with reference solution (b). The
discernible at 1190 cm-1. test is not valid unless the chromatogram obtained with
B. When examined in the range 210 nm to 360 nm, a 0.0008 per reference solution (c) shows a clearly visible spot.
cent w/v solution in 1 M sulphuric acid shows an absorption Heavy metals (2.3.13). 1.0 g complies with the limit test for
maximum at about 218 nm and a minimum at about 260 nm heavy metals, Method B (20 ppm).
(2.4.7).
C. In the test for Related substances, the principal spot in the
(d). Assay. Weigh accurately about 0.25 g and dissolve in 75 ml of
D. Dissolve about 1 mg in a mixture of 1 ml of ethanol and 5 ml
of a freshly prepared 2 per cent w/v solution of citric acid in
acetic anhydride. Heat in a water-bath for 10 to 15 minutes; a
reddish violet colour is produced. 1 ml of 0.1 M perchloric acid is equivalent to 0.02523 g of
C10H16N6S.
Tests
Mobile phase (a). A mixture of 65 volumes of ethyl acetate, Cimetidine Tablets
20 volumes of methanol and 15 volumes of strong ammonia Cimetidine Tablets contain not less than 95.0 per cent and not
solution. more than 105.0 per cent of the stated amount of cimetidine,
Mobile phase (b). A mixture of 84 volumes of ethyl acetate, C10H16N6S.
solution. Identification
Test solution (a). Dissolve 0.5 g of the substance under A. Shake a quantity of the powdered tablets containing 0.1 g
examination in sufficient methanol to produce 10 ml. of Cimetidine with 10 ml of methanol, filter, evaporate the
314
IP 2007 CINNARIZINE
filtrate to dryness using gentle heat and dry the residue at 60° test is not valid unless the chromatogram obtained with
at a pressure not exceeding 0.7 kPa. The residue complies with reference solution (c) shows a clearly visible spot.
the following test. Other tests. Comply with the tests stated under Tablets.
Compare the spectrum with that obtained with cimetidine RS
quantity of the powder containing about 0.25 g of Cimetidine
or with the reference spectrum of cimetidine.
and stir with 20 ml of warm methanol. Filter and repeat the
B. In the test for Related substances, the principal spot in the extraction with three quantities, each of 20 ml, of warm
chromatogram obtained with test solution (b) corresponds to methanol. Evaporate the combined filtrate and washings to
that in the chromatogram obtained with reference solution dryness and dissolve the residue in 75 ml of anhydrous glacial
(d). acetic acid. Titrate with 0.1 M perchloric acid, determining
Tests titration.
Related substances. Determine by thin-layer chromatography 1 ml of 0.1 M perchloric acid is equivalent to 0.02523 g of
(2.4.17), coating the plate with silica gel GF254. C10H16N6S.
Mobile phase (a). A mixture of 65 volumes of ethyl acetate,

solution.
Cinnarizine
Mobile phase (b). A mixture of 84 volumes of ethyl acetate,
8 volumes of methanol and 8 volumes of strong ammonia H
solution.
Test solution (a). Add 20 ml of methanol to a quantity of the N
powdered tablets containing 1 g of Cimetidine, mix with the N H
aid of ultrasound for 2 minutes, shake for 3 minutes and filter
using a suitable 0.2 µm filter.
methanol.
Reference solution (a). Dilute 1 ml of test solution (a) to C26H28N2 Mol. Wt. 368.5
100 ml with methanol and dilute 20 ml of this solution to 100 ml Cinnarizine is (E)-1-(diphenylmethyl)-4-(3-phenylprop-2-
with methanol. enyl)piperazine.
Reference solution (b). Dilute 5 ml of reference solution (a) to Cinnarizine contains not less than 99.0 per cent and not more
10 ml with methanol. than 101.0 per cent of C26H28N2, calculated on the dried basis.
Reference solution (c). Dilute 5 ml of reference solution (b) to Description. A white or almost white powder.
Reference solution (d). Dissolve 10 mg of cimetidine RS in
Identification
2 ml of methanol. Test A may be omitted if tests B and C are carried out. Tests B
Apply separately to two plates 4 µl of each solution. Allow and C may be omitted if test A is carried out.
the first plate to stand for 15 minutes in the tank saturated A. Determine by infrared absorption spectrophotometry (2.4.6).
with vapour from mobile phase (a). Develop the second plate Compare the spectrum with that obtained with cinnarizine
using mobile phase (b). After development, dry the plates in a RS.
current of air, expose to iodine vapour until maximum contrast
B. In the test for Related substances the principal spot in the
of the spots has been obtained and examine in ultraviolet light
at 254 nm. The following limits apply to both methods. Any
secondary spot in the chromatogram obtained with test
solution (a) is not more intense than the spot in the C. Dissolve 0.2 g of anhydrous citric acid in 10 ml of acetic
chromatogram obtained with reference solution (a) and not anhydride in a water-bath at 80° and maintain the temperature
more than two such spots are more intense than the spot in of the water-bath at 80° for 10 minutes. Add about 20 mg of the
the chromatogram obtained with reference solution (b). The substance under examination; a purple colour is produced.
315
CINNARIZINE TABLETS IP 2007
Tests 1 ml of 0.1 M perchloric acid is equivalent to 0.01843 of

C26H28N2.
dichloromethane is clear (2.4.1) and not more intensely
coloured than reference solution BYS6 (2.4.1).
Acidity or Alkalinity. Suspend 0.5 g in 15 ml of water. Boil for
2 minutes, cool and filter. Dilute the filtrate to 20 ml with carbon
Cinnarizine Tablets
dioxide-free water. To 10 ml add 0.1 ml of phenolphthalein Cinnarizine tablets contain not less than 90.0 per cent and not
solution and 0.25 ml of 0.01 M sodium hydroxide; the solution more than 110.0 per cent of the stated amount of cinnarizine,
is pink. To 10 ml add 0.1 ml of methyl red solution and 0.25 ml C26H28N2.
of 0.01 M hydrochloric acid; the solution is red.
Related substances. Determine by thin-layer chromatography Identification
(2.4.17), coating the plate with silica gel GF 254. A. Extract a quantity of the powdered tablets containing 0.1 g
Mobile phase. A mixture of 90 volumes of toluene and of Cinnarizine with 20 ml of dichloromethane, filter and
10 volumes of methanol. evaporate the filtrate to dryness. The residue complies with
the following test.
examination in 10 ml of dichloromethane. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with cinnarizine
RS.
dichloromethane.
cinnarazine RS in dichloromethane.
substance under examination in dichloromethane. Tests
Apply to the plate 10 µl of each solution. After development, Related substances. Determine by liquid chromatography
dry the plate in air, heat at 105° for 5 minutes and examine in (2.4.14).
ultraviolet light at 254 nm. Expose the plate to iodine vapours
for 15 minutes and examine in daylight. Any secondary spot Test solution (a). Shake a suitable quantity of the powdered
in the chromatogram obtained with test solution (a) is not tablets containing 25 mg of Cinnarizine with methanol, dilute
more intense than the spot in the chromatogram obtained to 10 ml with the same solvent and filter.
with reference solution (b). Reference solution (a). Dissolve 12.5 mg of cinnarizine RS
Heavy metals (2.3.13). Dissolve 1.0 g in a mixture of 85 volumes and 15 mg of flunarizine hydrochloride RS in methanol and
of acetone and 15 volumes of water and add dilute dilute to 100 ml with the same solvent. Dilute 1 ml of this
hydrochloric acid until dissolution is complete. Dilute to solution to 20 ml with methanol.
20 ml with the same mixture of acetone and water. 12 ml of the Reference solution (b). Dilute 1 ml of the test solution to 100
resulting solution complies with the limit test for heavy metals, ml with methanol. Dilute 5 ml of this solution to 20 ml with
Method D (20 ppm). Prepare the standard using 10 ml of lead methanol.
standard solution (1 ppm Pb) obtained by diluting lead
standard solution (100 ppm Pb) with the mixture of acetone Chromatographic system
and water. – a stainless steel column 10 cm × 4 mm, packed with
base-deactivated octadecylsilyl silica gel (3 µm),
Sulphated ash (2.3.18). Not more than 0.1 per cent. – mobile phase: A. a 10 per cent w/v solution of
Loss on drying (2.4.19). Not more than 0.5 per cent, determined ammonium acetate,
on 1.0 g by drying in an oven at 60° at a pressure not exceeding B. a 0.2 per cent v/v solution of glacial
0.7 kPa for 4 hours. acetic acid in acetonitrile,
of 70 volumes of 2-butanone and 10 volumes of anhydrous – a linear gradient programme using the conditions given
glacial acetic acid. Titrate with 0.1 M perchloric acid, using below,
á-naphtholbenzein solution as indicator. Carry out a blank – spectrophotometer set at 230 nm,
titration. – a 10 µl loop injector.
316
IP 2007 CIPROFLOXACIN
Time Mobile Mobile Comment Ciprofloxacin

phase A phase B
0 – 20 75 →10 25 → 90 linear gradient HN
N N
20 – 25 10 90 isocratic elution
25 – 30 75 25 switch to initial
F COOH
eluent composition
O
30 = 0 75 25 restart gradient
C17H18FN3O3 Mol. Wt. 331.4
Equilibrate the column for at least 30 minutes at the initial
eluent composition Ciprofloxacin is 1-cyclopropyl-6-fluoro-1,4-dihydro-4- oxo-7-
(piperazin-1-yl)quinoline-3-carboxylic acid.
Inject reference solution (b). Adjust the sensitivity of the
Ciprofloxacin contains not less than 98.0 per cent and not
system so that the height of the principal peak in the
more than 102.0 per cent of C17H18FN3O3, calculated on the
chromatogram obtained is at least 50 per cent of the full scale dried basis.
of the recorder. If necessary, adjust the concentration of glacial
acetic acid in mobile phase B to obtain a horizontal base-line Description. A white to pale yellow, crystalline powder.
in the chromatogram.
Identification
Inject reference solution (a). When the chromatogram is
recorded in the prescribed conditions, the retention times are: A. Determine by infrared absorption spectrophotometry (2.4.6).
cinnarizine about 11 min and flunarizine about 11.5 min. The Compare the spectrum with that obtained from ciprofloxacin
test is not valid unless the resolution between the peaks RS or with the reference spectrum of ciprofloxacin.
corresponding to cinnarizine and flunarizine is at least 5.0. B. Determine by thin-layer chromatography (2.4.17), coating
If necessary, adjust the time programme for the gradient the plate with silica gel G.
elution. Mobile phase. A mixture of 40 volumes of dichloromethane,
Inject the blank, the test solution and reference solution (b). 40 volumes of methanol, 20 volumes of strong ammonia
In the chromatogram obtained with the test solution: the area solution and 10 volumes of acetonitrile.
of any peak, other than the principal peak, is not greater than Test solution. Dissolve 0.1 g of the substance under
the area of the principal peak in the chromatogram obtained examination in 10 ml of 6 M ammonia.
with reference solution (b) (0.25 per cent); the sum of the Reference solution. A 1 per cent w/v solution of ciprofloxacin
areas of the peaks, other than the principal peak, is not greater RS in 6 M ammonia.
than twice the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.5 per cent). Ignore any Apply to the plate, as 1-cm bands, 5 µl of each solution. Place
peak due to the blank and any peak with an area less than the plate in an atmosphere of ammonia for about 15 minutes
0.2 times the area of the principal peak in the chromatogram and transfer it to an unsaturated chamber containing the
obtained with reference solution (b). mobile phase. Allow the mobile phase to rise 12 cm. Dry the
plate in air for 15 minutes and examine in ultraviolet light at
Other tests. Comply with the tests stated under tablets. 254 nm and at 365 nm. The principal band in the chromatogram
Assay. Determine by liquid chromatography (2.4.14) as given obtained with the test solution corresponds to that in the
under the test for Related substances using the following chromatogram obtained with the reference solution.
solutions. Tests
Test solution. Weigh and powder 20 tablets. Shake a quantity
Appearance of solution. A 2.5 per cent w/v solution in 0.1 M
of the powdered tablets containing about 25 mg of Cinnarazine
hydrochloric acid is clear (2.4.1).
with methanol, dilute to 50.0 ml with the same solvent and
filter. Dilute 5.0 ml of this solution to 50.0 ml with methanol. Related substances. Carry out the method described in the
Assay and calculate the percentage of each impurity from the
Reference solution. A 0.005 per cent w/v solution of cinnarzine chromatogram obtained with the test solution. The content of
RS in methanol. ciprofloxacin ethylenediamine analog or of any other
Calculate the content of C26H28N2 in the tablets. individual impurity peak found is not more than 0.2 per cent
and the sum of all the impurity peaks is not more than 0.5 per
Storage. Store protected from light. cent.
317
CIPROFLOXACIN INJECTION IP 2007
Fluoroquinolonic acid. Determine by thin-layer triethylamine to a pH of 3.0 ± 0.1, and 13 volumes of

chromatography (2.4.17), coating the plate with silica gel acetonitrile,
GF254. – flow rate. 1.5 ml per minute,
Mobile phase. A mixture of 40 volumes of dichloromethane, – column temperaure. 30° ± 1°,
40 volumes of methanol, 20 volumes of strong ammonia – spectrophotometer set at 278 nm,
solution and 10 volumes of acetonitrile. – a 10 µl loop injector.
Test solution. Dissolve 0.1 g of the substance under Inject reference solution (b) and record the chromatogram
examination in 10 ml of 0.1 M acetic acid. adjusting the sensitivity and flow rate suitably so that the
retention time for ciprofloxacin is between 6.4 and 10.8 minutes,
Reference solution. Weigh 10 mg of fluoroquionolonic acid the relative retention times are about 0.7 for ciprofloxacin
RS, add 0.1 ml of 6 M ammonia and dilute to 100.0 ml with ethylenediamine analog and 1.0 for ciprofloxacin and the
water. Dilute 2.0 ml of this solution to 10.0 ml with water. resolution between ciprofloxacin ethylenediamine analog peak
Apply to the plate 5 µl of each solution. Place the plate in an and ciprofloxacin peak is not less than 6. The column efficiency,
atmosphere of ammonia for about 15 minutes. Remove the determined from ciprofloxacin peak, is not less than
plate and place it in a chamber containing the mobile phase. 2500 theoretical plates, the tailing factor for the ciprofloxacin
After development, dry the plate in air for 15 minutes and peak is not more than 4.0 and the relative standard deviation
examine in ultraviolet light at 254 nm. Any secondary spot in for replicate injections is not more than 1.5 per cent.
the chromatogram obtained with the test solution
corresponding to the spot of fluoroquionolonic acid is not
more intense than the spot in the chromatogram obtained Calculate the content of C17H18FN3O3.
with the reference solution. Storage. Store protected from light.
Chlorides (2.3.12). To 2.0 g add 30 ml of water, shake for
5 minutes and filter through a chloride-free filter paper. 15 ml Ciprofloxacin Injection
of the filtrate complies with the limit test for chlorides Ciprofloxacin Injection is a sterile solution of Ciprofloxacin in
(250 ppm). 5 per cent Dextrose Injection or in Sodium Chloride Injection
Sulphates (2.3.17). Dissolve 0.75 g in 5.0 ml of 2 M acetic acid prepared with the aid of Lactic Acid.
and 20.0 ml of water.10 ml of the resulting solution complies Ciprofloxacin Injection contains not less than 90.0 per cent
with the limit test for sulphates (400 ppm). and not more than 110.0 per cent of the stated amount of
Sulphated ash (2.3.18). Not more than 0.1 per cent. ciprofloxacin, C17H18FN3O3.
Identification
on 1.0 g by drying in an oven at 120° for 6 hours at a pressure
not exceeding 0.7 kPa. Determine by thin-layer chromatography (2.4.17), coating the
Assay. Determine by liquid chromatography (2.4.14). plate with silica gel G. Place the plate in an atmosphere of
ammonia for about 15 minutes and transfer it to an unsaturated
Test solution. Weigh accurately about 25 mg, add 0.2 ml of a chamber.
solution containing 7 per cent v/v of phosphoric acid and
add sufficient of the mobile phase to produce 50.0 ml. Mobile phase. A mixture of 40 volumes of dichloromethane,
40 volumes of methanol, 20 volumes of strong ammonia
Reference solution (a).Prepare in the same manner as the test solution and 10 volumes of acetonitrile.
solution using an accurately weighed quantity of
ciprofloxacin RS in place of the substance under examination. Test solution. Dilute sufficient of the injection with water to
obtain a solution containing the equivalent of 0.05 per cent
Reference solution (b). A 0.05 per cent w/v solution of w/v of Ciprofloxacin.
ciprofloxacin ethylenediamine analog RS in reference
solution (a). Reference solution. A 0.05 per cent w/v solution of
ciprofloxacin RS in 6 M ammonia.
– a stainless steel column 25 cm × 4 mm, packed with Apply to the plate, as 1-cm bands, 5 µl of each solution. Place
octadecylsilyl silica gel (5 µm), the plate in an atmosphere of ammonia for about 15 minutes
– mobile phase: a mixture of 87 volumes of 0.025 M and transfer it to an unsaturated chamber containing the
phosphoric acid, previously adjusted with mobile phase. Allow the mobile phase to rise 12 cm. Dry the
318
IP 2007 CIPROFLOXACIN INJECTION
plate in air for 15 minutes and examine in ultraviolet light at add 0.2 ml of 6 M ammonia and dilute to 100.0 ml. Mix well and
254 nm and at 365 nm. The principal band in the chromatogram determine the optical rotation at 25° in a 2-dm tube (2.4.22).
obtained with the test solution corresponds to that in the The observed rotation in degrees multiplied by 2.085 represents
chromatogram obtained with the reference solution. the percentage of dextrose monohydrate, C6H12O6,H2O, in the
preparation under examination.
Tests
Sodium chloride (if present). 0.855 per cent to 0.945 per cent
pH (2.4.24). 3.5 to 4.6. w/v of NaCl, determined by the following method. To 10.0 ml
Ciprofloxacin ethylenediamine analog. Not more than 0.5 per add 150 ml of water and titrate with 0.1 M silver nitrate using
cent, determined by the method described in the Assay. potassium chromate solution as indicator.
Calculate the percentage of ciprofloxacin ethylenediamine 1 ml of 0.1 M silver nitrate is equivalent to 0.005844 g of NaCl.
analog from the chromatogram obtained with the test solution
from the following expression.
Unit per mg of ciprofloxacin.
Per cent of the analog = 100[0.7 × ra /(0.7 x ra + rc)], where 0.7 is
the response factor for ciprofloxacin ethylenediamine analog Sterility (2.2.11). Complies with the test for sterility, using
relative to that of ciprofloxacin, ra and rc are the responses of Method A.
ciprofloxacin ethylenediamine analog peak and the Particulate contamination (2.5.9). Complies with the limit test
ciprofloxacin peak respectively. for particulate contamination.
Lactic acid. 0.288 mg to 0.352 mg for each mg of Ciprofloxacin Other tests. Complies with the tests stated under Parenteral
stated on the label. Preparations (Injections).
Determine by liquid chromatography (2.4.14). Assay. Deteremine by liquid chromatography (2.4.14).
Test solution. The substance under examination. Test solution. Dilute a volume of the injection containing
Reference solution. A 0.08 per cent w/v solution of sodium 25 mg of Ciprofloxacin to 100.0 ml with the mobile phase and
lactate RS in water. mix.
Chromatographic system Reference solution (a). A 0.03 per cent w/v solution of
– a stainless steel column 30 cm × 7.8 mm, packed with a ciprofloxacin hydrochloride RS in the mobile phase.
strong cation-exchange resin consisting of sulphonated
Reference solution (b). Dissolve a sufficient quantity of
cross-linked styrene-divinylbenzene copolymer in the
ciprofloxacin ethylenediamine analog RS in reference
hydrogen form (7 to 11 µm),
solution (a) so as to obtain a solution containing 0.025 per
– mobile phase: a mixture of 85 volumes of 0.0025 M
cent w/v of the reference substance.
sulphuric acid and 15 volumes of acetonitrile,
– column temperature. 40° ± 1°, Chromatographic system
– flow rate. 0.6 ml per minute, – a stainless steel column 25 cm x 4 mm, packed with
– spectrophotometer set at 208 nm, octadecylsilyl silica gel (5 µm),
– a 20 µl loop injector. – mobile phase: a mixture of 87 volumes of 0.025 M
phosphoric acid, previously adjusted with triethyl-
Inject the reference solution and record the chromatograms
amine to a pH of 3.0 ± 0.1, and 13 volumes of acetonitrile,
adjusting the sensitivity and flow rate suitably so that the
– column temperaure. 30° ± 1°,
Inject alternately the test solution and reference solution, – a 10 µl loop injector.
record the chromatograms and measure the peak responses
for the major peaks. Calculate the content of lactic acid, C3H6O3, Inject reference solution (b) and record the chromatogram
in the substance under examination. adjusting the sensitivity and flow rate suitably so that the
NOTE - After each analysis, the column should be rinsed the relative retention times are about 0.7 for ciprofloxacin
with a mixture of 85 volumes of 0.005 M sulphuric acid and ethylenediamine analog and 1.0 for ciprofloxacin and the
15 volumes of acetonitrile to elute the ciprofloxacin from the resolution between ciprofloxacin ethylenediamine analog peak
column. The column may be regenerated with 0.005 M and ciprofloxacin peak is not less than 6. The column efficiency,
sulphuric acid and may be reused or stored. determined from ciprofloxacin peak, is not less than
Dextrose (if present). 4.75 per cent to 5.25 per cent w/v of 2500 theoretical plates, the tailing factor for the ciprofloxacin
C6H12O6,H2O, determined by the following method. To 50.0 ml peak is not more than 4.0 and the relative standard deviation
319
CIPROFLOXACIN HYDROCHLORIDE IP 2007
for replicate injections is not more than 1.5 per cent. Related substances. Carry out the method described in the
Inject alternately the test solution and reference solution (a). Assay and calculate the percentage of each impurity peak in
the chromatogram obtained with the test solution. The content
Calculate the content of C17H18FN3O3 in the injection. of ciprofloxacin ethylenediamine analog or of any other
Storage. Store protected from light at a temperature not individual impurity peak found is not more than 0.2 per cent
exceeding 30°. The contents should not be allowed to freeze. and the sum of all the impurity peaks is not more than 0.5 per
cent.
Labelling. The label states whether Dextrose or Sodium
Chloride has been used for preparing the injection. Fluoroquinolonic acid. Determine by thin-layer
chromatography (2.4.17), coating the plate with silica gel
GF254.
Ciprofloxacin Hydrochloride 40 volumes of methanol, 20 volumes of strong ammonia
C17H18FN3O3,HCl,H2O Mol. Wt. 385.8 solution and 10 volumes of acetonitrile.
Ciprofloxacin Hydrochloride is 1-cyclopropyl-6-fluoro-1, 4- Test solution. Dissolve 0.1 g of the substance under
dihydro-4-oxo-7-(1-piperazinyl)-3-quinolinecarboxylic acid examination in 10 ml of water.
hydrochloride monohydrate. Reference solution. Weigh 10 mg of fluoroquionolonic acid
Ciprofloxacin Hydrochloride contains not less than 98.0 per RS, add 0.1 ml of 6 M ammonia and dilute to 100.0 ml with
cent and not more than 102.0 per cent of C17H18FN3O3,HCl, water. Dilute 2.0 ml of this solution to 10.0 ml with water.
calculated on the anhydrous basis. Apply to the plate 5 µl of each solution. Place the plate in an
atmosphere of ammonia for about 15 minutes. Remove the
Description. A pale yellow, crystalline powder.
plate and place it in a chamber containing the mobile phase.
Identification After development, dry the plate in air for 15 minutes and
A. Determine by infrared absorption spectrophotometry (2.4.6). the chromatogram obtained with the test solution
Compare the spectrum with that obtained with ciprofloxacin corresponding to the spot of fluoroquionolonic acid is not
hydrochloride RS. more intense than the spot in the chromatogram obtained
B. Determine by thin-layer chromatography (2.4.17), coating with the reference solution.
the plate with silica gel G. Place the plate in an atmosphere of Heavy metals (2.3.13). 1.0 g complies with the limit test for
ammonia for about 15 minutes and transfer it to an unsaturated heavy metals, Method B (20 ppm).
chamber.
Sulphates (2.3.17). 0.375 g complies with the limit test for
Mobile phase. A mixture of 40 volumes of dichloromethane, sulphates (400 ppm).
40 volumes of methanol, 20 volumes of strong ammonia Sulphated ash (2.3.18). Not more than 0.1 per cent.
solution and 10 volumes of acetonitrile.
examination in 10 ml of water.
Test solution. Weigh accurately about 50 mg of the substance
Reference solution. A 1 per cent w/v solution of ciprofloxacin under examination and dissolve in 100.0 ml of water.
hydrochloride RS in water.
Apply to the plate, as 1-cm bands, 5 µl of each solution. Place ciprofloxacin hydrochloride RS in water.
the plate in an atmosphere of ammonia for about 15 minutes
and transfer it to an unsaturated chamber containing the
ciprofloxacin ethylenediamine analog RS in water.
mobile phase. Allow the mobile phase to rise 12 cm. Dry the
plate in air for 15 minutes and examine in ultraviolet light at Chromatographic system
254 nm and at 365 nm. The principal band in the chromatogram – a stainless steel column 25 cm x 4 mm, packed with
obtained with the test solution corresponds to that in the octadecylsilyl silica gel (5 µm),
chromatogram obtained with the reference solution. – mobile phase: a mixture of 87 volumes of 0.025 M
phosphoric acid, previously adjusted with
C. Gives the reactions of chlorides (2.3.1).
triethylamine to a pH of 3.0 ± 0.1, and 13 volumes of
Tests acetonitrile,
pH (2.4.24). 3.0 to 4.5, determined in a 2.5 per cent w/v solution. – column temperaure. 30° ± 1°,
320
IP 2007 CIPROFLOXACIN TABLETS
– spectrophotometer set at 278 nm, – mobile phase: a mixture of 75 volumes of 0.005 M

– a 10 µl loop injector. tetrabutylammonium phosphate, adjusted to pH 2.0 with
Inject reference solution (b) and record the chromatogram orthophosphoric acid and 25 volumes of methanol,
adjusting the sensitivity and flow rate suitably so that the – flow rate. 1.5 ml per minute,
retention time for ciprofloxacin is between 6.4 and 10.8 minutes, – spectrophotometer set at 280 nm,
the relative retention times are about 0.7 for ciprofloxacin – a 20 µl loop injector.
ethylenediamine analog and 1.0 for ciprofloxacin and the Inject reference solution (b). The relative retention time are
resolution between ciprofloxacin ethylenediamine analog peak about 0.8 for the ciprofloxacin ethylenediamine analog and 1.0
and ciprofloxacin peak is not less than 6. The column efficiency, for ciprofloxacin and the resolution between the ciprofloxacin
determined from ciprofloxacin peak, is not less than ethylenediamine analog peak and the ciprofloxacin peak is
2500 theoretical plates, the tailing factor for the ciprofloxacin not less than 1.5.
peak is not more than 4.0 and the relative standard deviation Inject reference solution (a). The test is not valid unless the
for replicate injections is not more than 1.5 per cent. column efficiency is not less than 500 theoretical plates, the
Inject alternately the test solution and reference solution (a). tailing factor not more than 2.0, and the relative standard
deviation for replicate injections is not more than 2 per cent.
Calculate the content of C17H18FN3O3,HCl.
Inject alternatively the test solution and reference solution (a).
Calculate the content of C17H18FN3O3 in the eye drops.
Ciprofloxacin Eye Drops
Ciprofloxacin Eye Drops are a sterile solution of Ciprofloxacin
Hydrochloride in Purified water.
Ciprofloxacin Tablets
Ciprofloxacin Eye Drops contain not less than 90.0 per cent Ciprofloxacin Hydrochloride Tablets
and not more than 110.0 per cent of the stated amount of Ciprofloxacin Tablets contain not less than 90.0 per cent and
ciprofloxacin, C17H18FN3O3. not more than 110.0 per cent of the stated amount of
ciprofloxacin, C17H18FN3O3. The tablets may be coated.
Identification
Identification
obtained with the test solution corresponds to the peak in the A. In the Assay, the principal peak in the chromatogram
chromatogram obtained with the reference solution. obtained with the test solution corresponds to the peak in the
B. Give reaction A of chlorides (2.3.1). chromatogram obtained with the reference solution.
Tests the plate with silica gel G. Place the plate in an atmosphere of
pH (2.4.24). 3.5 to 5.5 ammonia for about 15 minutes and transfer it to an unsaturated
chamber.
Other tests. Comply with the tests stated under Eye Drops.
40 volumes of methanol, 20 volumes of strong ammonia
Test solution. Transfer an accurately measured volume of Eye solution and 10 volumes of acetonitrile.
drops containing 6 mg of ciprofloxacin, to a 50-ml volumetric
flask, dilute with water to volume, and mix.
containing about 0.15 g of ciprofloxacin with 75 ml of water
Reference solution (a). A 0.014 per cent w/v solution of for 20 minutes, dilute to 100.0 ml with water, mix, centrifuge
ciprofloxacin hydrochloride RS in water. and use the clear supernatant liquid.
Resolution solution (b). A 0.001 per cent w/v solution of Reference solution. A 0.15 per cent w/v solution of
ciprofloxacin ethylenediamine analog RS in water ciprofloxacin hydrochloride RS in water.
Chromatographic system Apply to the plate, as 1-cm bands, 5 µl of each solution. Place
– a stainless steel column 25 cm x 4.6 mm packed with the plate in an atmosphere of ammonia for about 15 minutes
octadecylsilane bonded to porous silica (5 µm), and transfer it to an unsaturated chamber containing the
– column temperature 30º, mobile phase. Allow the mobile phase to rise 12 cm. Dry the
321
CISPLATIN IP 2007
plate in air for 15 minutes and examine in ultraviolet light at and ciprofloxacin peak is not less than 6. The column efficiency,
254 nm and at 365 nm. The principal band in the chromatogram determined from ciprofloxacin peak, is not less than
obtained with the test solution corresponds to that in the 2500 theoretical plates, the tailing factor for the ciprofloxacin
chromatogram obtained with the reference solution. peak is not more than 4.0 and the relative standard deviation
Tests Inject alternately the test solution and reference solution (a).
Dissolution (2.5.2). Calculate the content of C17H18FN3O3,HCl in the tablets.
Apparatus. No 1 Storage. Store protected from light.
Medium. 900 ml of water Labelling. The label states the strength in terms of the
Speed and time. 50 rpm and 30 minutes. equivalent amount of ciprofloxacin.
the absorbance of the filtrate, suitably diluted with water if
necessary, at the maximum at about 276 nm (2.4.7). Calculate Cisplatin
the content of ciprofloxacin, C17H18FN3O3, in the medium from
the absorbance obtained by repeating the determination using NH3
Cl
a solution of known concentration of ciprofloxacin Pt
hydrochloride RS. Cl NH3
D. Not less than 80 per cent of the stated amount of H6Cl2N2Pt Mol. Wt. 300.0
C17H18FN3O3.
Cisplatin is cis-diamminedichloroplatinum(II).
Cisplatin contains not less than 97.0 per cent and not more
Assay. Determine by liquid chromatography (2.4.14). than 102.0 per cent of H6Cl2N2Pt.
Test solution. Weigh and powder 20 tablets. Weigh accurately Description. A yellow powder or orange yellow crystals.
a quantity of the powder containing about 1.25 g of
CAUTION - Cisplatin is potentially cytotoxic. Great care
ciprofloxacin, add about 400 ml of 0.01 M hydrochloric acid,
should be taken in handling the powder and preparing
shake for 20 minutes, dilute to 500.0 ml with 0.01 M
solutions.
hydrochloric acid. and filter. Dilute 10.0 ml of the filtrate to
100.0 ml with 0.01 M hydrochloric acid. NOTE - Carry out all the tests and the Assay, except
Identification tests A and C and the test for Silver, protected
from light.
ciprofloxacin hydrochloride RS in 0.01 M hydrochloric acid.
Reference solution (b). A 0.05 per cent w/v solution of Identification
ciprofloxacin ethylenediamine analog RS in water.
Test A may be omitted if tests B and C are carried out. Test C
Chromatographic system may be omitted if tests A and B are carried out.
Compare the spectrum with that obtained with cisplatin RS.
phosphoric acid, previously adjusted with B. In the test for Related substances, the principal spot in the
triethylamine to a pH of 3.0 ± 0.1, and 13 volumes of chromatogram obtained with test solution (b) corresponds to
acetonitrile, that in the chromatogram obtained with reference solution (b).
– flow rate. 1.5 ml per minute, C. Add 50 mg to 2 ml of 2 M sodium hydroxide, evaporate to
– column temperaure. 30° ± 1°, dryness, dissolve the residue in a mixture of 0.5 ml of nitric
– spectrophotometer set at 278 nm, acid and 1.5 ml of hydrochloric acid and evaporate to dryness
– a 10 µl loop injector. again; the residue is orange. Dissolve the residue in 0.5 ml of
Inject reference solution (b) and record the chromatogram water and add 0.5 ml of ammonium chloride solution; a yellow
adjusting the sensitivity and flow rate suitably so that the crystalline precipitate is produced.
Tests
the relative retention times are about 0.7 for ciprofloxacin
ethylenediamine analog and 1.0 for ciprofloxacin and the Appearance of solution. Solution A is clear (2.4.1) and not
resolution between ciprofloxacin ethylenediamine analog peak more intensely coloured than reference solution GYS5 or BYS5
322
IP 2007 CISPLATIN INJECTION
(2.4.1). A 2.0 per cent w/v solution in dimethylformamide is – mobile phase: a mixture of 90 volumes of methanol and
clear (2.4.1). 10 volumes of normal saline
in normal saline prepared in carbon dioxide-free water
(solution A), measured immediately after preparation
Calculate the content of H6Cl2N2Pt.
(2.4.17), coating the plate with microcrystalline cellulose and Storage. Store protected from light.
activating the plate by heating at 150° for 1 hour.
Mobile phase. A mixture of 90 volumes of dimethylformamide
and 10 volumes of acetone. Cisplatin Injection
Test solution (a). A 2 per cent w/v solution of the substance Cisplatin Injection is a sterile, freeze-dried mixture of Cisplatin,
under examination in dimethylformamide. Mannitol and Sodium Chloride. It is filled in a sealed container.
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with The injection is constituted by dissolving the contents of the
dimethylformamide. sealed container in the requisite amount of sterile Water for
Reference solution (a). Dilute 5 ml of test solution (b) to 25 ml Injections, immediately before use.
with dimethylformamide. The constituted solution complies with the requirements for
Reference solution (b). A 0.2 per cent w/v solution of cisplatin Clarity of solution and Particulate matter stated under
RS in dimethylformamide. Parenteral Preparations (Injections).
Apply to the plate 5 µl of each solution. After development, Storage. The constituted solution should be used immediately
dry the plate in air and spray with a 5 per cent w/v solution of after preparation but, in any case, within the period
stannous chloride in 1 M hydrochloric acid. After 1 hour, the recommended by the manufacturer.
chromatogram obtained with the test solution shows no Cisplatin Injection contains not less than 90.0 per cent and
secondary spot with an Rf value lower than that of the principal not more than 110.0 per cent of the stated amount of cisplatin,
spot and any secondary spot with an Rf value higher than H6Cl2N2Pt.
that of the principal spot is not more intense than the spot in
the chromatogram obtained with reference solution (b). Description. A yellow powder.
Silver. Determine by atomic absorption spectrophotometry The contents of the sealed container comply with the
(2.4.2), measuring at 328 nm using a silver hollow-cathode requirements stated under Parenteral Preparations
light as a radiation source, a fuel-lean air-acetylene flame, (Powders for Injection) and with the following requirements.
preferably a spectral width of 0.5 nm. CAUTION — Cisplatin is potentially cytotoxic. Great care
Test solution. Dissolve 0.1 g of the substance under should be taken in handling the powder and preparing
examination in 15 ml of nitric acid by heating up to 80°. Cool solutions.
and dilute to 25.0 ml with water.
Identification
Reference solutions. Add 50 ml of nitric acid to suitable
volumes (10 to 30 ml) of silver solution AAS and dilute to A. When examined in the range 230 nm to 360 nm, a 0.1 per
100.0 ml with water. cent w/v solution in 0.1 M hydrochloric acid shows an
Carry out a blank determination (250 ppm). absorption maximum only at about 300 nm (2.4.7).
Assay. Determine by liquid chromatography (2.4.14). B. In the test for Related substances, the principal spot in the
Test solution. Prepare immediately before use a 0.05 per cent that in the chromatogram obtained with reference solution (b).
w/v solution of the substance under examination in normal
saline. Tests
Reference solution. A 0.05 per cent w/v solution of cisplatin pH (2.4.24). 3.5 to 6.5, determined in a solution constituted as
RS in normal saline. directed in the label, in water for injections.
Chromatographic system Related substances. Determine by thin-layer chromatography
– a stainless steel column 25 cm x 4.6 mm, packed with (2.4.17), coating the plate with microcrystalline cellulose and
strong anion-exchange silica gel (10 µm), activating the plate by heating at 150° for 1 hour.
323
CITRIC ACID IP 2007
Mobile phase. A mixture of 90 volumes of dimethylformamide Citric Acid Monohydrate contains not less than 99.0 per cent
and 10 volumes of acetone. and not more than 101.0 per cent of C6H8O7, calculated on the
Test solution. Shake the contents of one vial with anhydrous basis.
dimethylformamide to produce a solution containing 0.5 per Description. Colourless crystals or a white powder; slightly
cent w/v of Cisplatin, mix with the aid of ultrasound for hygroscopic in moist dry air.
10 minutes and filter.
Reference solution (a). Dilute 5 ml of the test solution to 50 ml Identification
with dimethylformamide. A. Gives reaction A of citrates (2.3.1).
Reference solution (b). Dilute 1 ml of the test solution to 50 ml B. A 10 per cent w/v solution is strongly acidic.
with dimethylformamide.
Reference solution (c). A solution containing 0.05 per cent Tests
w/v of cisplatin RS in dimethylformamide.
Appearance of solution. Dissolve 2.0 g in sufficient water to
Apply to the plate 20 µl of each solution. After development, produce 10 ml. The solution is clear (2.4.1), and not more
dry the plate in air and spray with a 5 per cent w/v solution of intensely coloured than reference solution YS7, BYS7 or GYS7
stannous chloride in 1 M hydrochloric acid. After 1 hour, the (2.4.1).
chromatogram obtained with the test solution shows no
secondary spot with an Rf value lower than that of the principal Arsenic (2.3.10). Dissolve 10.0 g in 50 ml of water and add
spot and any secondary spot with an Rf value higher than 10 ml of stannated hydrochloric acid; the resulting solution
that of the principal spot is not more intense than the spot in complies with the limit test for arsenic (1 ppm).
the chromatogram obtained with reference solution (b). Barium. Dissolve 5.0 g in several portions in 39 ml of 2 M
Bacterial endotoxins (2.2.3). Not more than 2.0 Endotoxin sodium hydroxide and dilute to 50 ml with distilled water
Units per mg of cisplatin. (solution A). To 5 ml of solution A add 5 ml of 1 M sulphuric
acid and allow to stand for 1 hour. Any opalescence produced
Sterility (2.2.11). Comply with the test for sterility, Method A.
is not more intense than that of a mixture of 5 ml of solution A
Assay. Determine by liquid chromatography (2.4.17). and 5 ml of distilled water.
Test solution. Determine the weight of the contents of Calcium. To 0.2 ml of ethanolic calcium standard solution
10 containers. Dissolve the mixed contents of 10 containers in (100 ppm Ca) add 1 ml of a 4 per cent w/v solution of
dimethylformamide to obtain a solution containing about ammonium oxalate. After 1 minute add 1 ml of 2 M acetic acid
0.1 per cent w/v of cisplatin. and 5 ml of solution A diluted to 10 ml with distilled water and
Reference solution. A 0.1 per cent w/v solution of cisplatin shake. After 15 minutes any opalescence produced is not more
RS in dimethylformamide (use within one hour). intense than that of a standard prepared in the same manner
Chromatographic system using a mixture of 10 ml of calcium standard solution
– a stainless steel column 25 cm x 4.6 mm, packed with (10 ppm Ca) and 5 ml of water in place of solution A (200
strong anion-exchange silica gel (10 µm), ppm).
– mobile phase: a mixture of 90 volumes of methanol and Heavy metals (2.3.13). 2.0 g complies with the limit test for
10 volumes of normal saline, heavy metals, Method A (10 ppm).
Iron (2.3.14). 8 ml of solution A diluted to 10 ml with water
complies with the limit test for iron (50 ppm).
Calculate the content of H6Cl2N2Pt in the injection. Chlorides (2.3.12). Dissolve 5.0 gm in 10 ml of water, add 1 ml
of 2 M nitric acid and dilute to 15 ml with water. The resulting
Storage. Store protected from light. solution complies with the limit test for chlorides. The resulting
solution complies with the limit test for chlorides (50 ppm).
Sulphates (2.3.17). Dissolve 1.0 g in sufficient distilled water
Citric Acid to produce 15 ml. The resulting solution complies with the
limit test for sulphates (150 ppm).
HO COOH
HOOC COOH Oxalic acid. Dissolve 0.8 g in 4 ml of water, add 2 ml of
hydrochloric acid and 1 g of granulated zinc and heat in a
C6H8O7 Mol. Wt. 192.1 water-bath for 1 minute. Allow to stand for 2 minutes, decant
Citric Acid is 2-hydroxypropane-1,2,3-tricarboxylic acid. the liquid into a test-tube containing 0.25 ml of a 1 per cent
324
IP 2007 CITRIC ACID MONOHYDRATE
w/v solution of phenylhydrazine hydrochloride and heat to (solution A). To 5 ml of solution A add 5 ml of 1 M sulphuric
boiling. Cool rapidly, transfer to a graduated measuring acid and allow to stand for 1 hour. Any opalescence produced
cylinder, add an equal volume of hydrochloric acid and is not more intense than that of a mixture of 5 ml of solution A
0.25 ml of a 5 per cent solution of potassium ferricyanide, and 5 ml of distilled water.
shake and allow to stand for 30 minutes. Any pink colour
Calcium. To 0.2 ml of ethanolic calcium standard solution
produced is not more intense than that produced by carrying
(100 ppm Ca) add 1 ml of a 4 per cent w/v solution of
out the test using 0.2 ml of oxalic acid dissolved in 4 ml of
ammonium oxalate. After 1 minute add 1 ml of 2 M acetic acid
water.
and 5 ml of solution A diluted to 10 ml with distilled water and
Readily carbonisable substances. Heat 0.75 g in powder, with shake. After 15 minutes any opalescence produced is not more
10 ml of sulphuric acid (containing 94.5 to 95.5 per cent w/w intense than that of a standard prepared in the same manner
of H2SO4) in a water-bath at 90°. Shake after one minute, using a mixture of 10 ml of calcium standard solution
continue the heating for a total of 1 hour and cool rapidly and (10 ppm Ca) and 5 ml of water in place of solution A (200
immediately. Any colour produced is not more intense than ppm).
that of a mixture of 1.0 ml of CCS and 9.0 ml of FCS (2.4.1).
Sulphated ash (2.3.18). Not more than 0.1 per cent. heavy metals, Method A (10 ppm).
Water (2.3.43). Not more than 1.0 per cent, determined on 2.0 g. Iron (2.3.14). 8 ml of solution A diluted to 10 ml with water
Assay. Weigh accurately about 2 g and dissolve in 100 ml of complies with the limit test for iron (50 ppm).
water. Titrate with 1 M sodium hydroxide using 0.5 ml of Chlorides (2.3.12). To 2 ml of solution A add 1 ml of 2 M nitric
phenolphthalein solution as indicator. acid and dilute to 15 ml with water. The resulting solution
1 ml of 1 M sodium hydroxide is equivalent to 0.06403 g of complies with the limit test for chlorides (50 ppm).
C6H8O7. Sulphates (2.3.17). Dissolve 1.0 g in sufficient distilled water
to produce 15 ml. The resulting solution complies with the
limit test for sulphates (150 ppm).
Citric Acid Monohydrate Oxalic acid. Dissolve 0.8 g in 4 ml of water, add 2 ml of
C6H8O7,H2O Mol. Wt. 210.1 hydrochloric acid and 1 g of granulated zinc and heat in a
water-bath for 1 minute. Allow to stand for 2 minutes, decant
Citric Acid Monohydrate is 2-hydroxypropane-1,2,3- the liquid into a test-tube containing 0.25 ml of a 1 per cent
tricarboxylic acid monohydrate. w/v solution of phenylhydrazine hydrochloride and heat to
Citric Acid Monohydrate contains not less than 99.0 per cent boiling. Cool rapidly, transfer to a graduated measuring
and not more than 101.0 per cent of C6H8O7, calculated on the cylinder, add an equal volume of hydrochloric acid and
anhydrous basis. 0.25 ml of a 5 per cent solution of potassium ferricyanide,
shake and allow to stand for 30 minutes. Any pink colour
Description. Colourless crystals or a white, crystalline powder; produced is not more intense than that produced by carrying
slightly efflorescent in warm, dry air. out the test using 0.2 ml of oxalic acid dissolved in 4 ml of
Identification water.
Readily carbonisable substances. Heat 0.50 g in powder, with
A. Gives reaction A of citrates (2.3.1).
5 ml of sulphuric acid (containing 94.5 to 95.5 per cent w/w of
B. A 10 per cent w/v solution is strongly acidic. H2SO4) in a water-bath at 90° in the dark. Shake after one
minute, continue heating for a total of 1 hour and cool rapidly
Tests and immediately. Any colour produced is not more intense
Appearance of solution. Dissolve 2.0 g in sufficient water to than that of a mixture of 0.6 ml of CCS and 5.4 ml of FCS (2.4.1).
produce 10 ml. The solution is clear (2.4.1), and not more Sulphated ash (2.3.18). Not more than 0.1 per cent.
intensely coloured than reference solution YS7, BYS7 or GYS7
(2.4.1). Water (2.3.43). 7.5 to 9.0 per cent, determined on 0.5 g.
Arsenic (2.3.10). Dissolve 10 g in 50 ml of water and add 10 ml Assay. Weigh accurately about 2 g and dissolve in 50 ml of
of stannated hydrochloric acid; the resulting solution water. Titrate with 1 M sodium hydroxide using 0.5 ml of
complies with the limit test for arsenic (1 ppm). phenolphthalein solution as indicator.
Barium. Dissolve 5.0 g in several portions in 39 ml of 2 M 1 ml of 1 M sodium hydroxide is equivalent to 0.06403 g of
sodium hydroxide and dilute to 50 ml with distilled water C6H8O7.
325
CLARITHROMYCIN IP 2007
Clarithromycin secondary peak is not more than 2.5 times the area of the peak
O not more than 5 times the area of the peak in the chromatogram
H3C CH3 obtained with the reference solution (b) (5.0 per cent).
H3C OH
CH3 HO Heavy metals (2.3.13). Dissolve 1 g in a mixture of 15 volumes
H3CO CH3
O O
CH3 of water and 85 volumes of dioxan and dilute to 20 ml with the
H3C CH3 O same mixture of solvents. 12 ml of the solution complies with
O CH3
N OH O
CH3 O
limit test for heavy metals, Method D (20 ppm). Prepare
OCH3 reference solution using 1 ml of lead standard solution (100
OH CH3
ppm) using the same mixture of solvents.
CH3
Water (2.3.43). Not more than 2.0 per cent w/w, determined on
0.5 g using pyridine as solvent.
C38H69NO13 Mol. Wt. 748.0 Assay. Determine by liquid chromatography (2.4.14).
Clarithromycin is (3R,4S,5S,6R,7R,9R,11R,12R,13S,14R)-4- Test solution. Dissolve 75 mg of the substance under
[(2,6-Dideoxy-3-C-methyl-3-O-methyl-á-L-ribo- examination in 25 ml of acetonitrile and dilute to 50.0 ml with
hexopyranosyl)oxy]-14-ethyl-12,13-dihydroxy-7-methoxy- water.
3,5,7,9,11,13-hexamethyl-6-[[3,4,6-trideoxy-3-(dimethylamino)-
Reference solution. Dissolve 15 mg of the clarithromycin RS
á ?D-xylo-hexopyranosyl]oxy]oxacyclotetradecane-2,10-dione
in 5 ml of acetonitrile and dilute to 10.0 ml with water.
(6-O-methylerythromycin A).
Clarithromycin contains not less than 96.0 per cent and not
more than 102.0 per cent of C38H69NO13, calculated on the
octadecylsilyl silica (3.5µm),
anhydrous basis.
Description. A white or almost white, crystalline powder. – mobile phase: A. 0.476 per cent w/v solution of
Identification potassium dihydrogen phosphate adjusted to pH 4.4
with dilute orthophosphoric acid or a 4.5 per cent
Determine by infrared absorption spectrophotometry (2.4.6). solution of potassium hydroxide, filter,
Compare the spectrum with that obtained with clarithromycin B. acetonitrile,
RS or with the reference spectrum of clarithromycin.
Tests below,
Specific optical rotation (2.4.22). -94º to -102º, determined on – spectrophotometer set at 205 nm,
a 1 per cent w/v solution in methylene chloride at 20º. – a 10 µl loop injector.
Related substances. Determine by liquid chromatography Time Mobile phase A Mobile phase B
(2.4.14). (in mins.) (per cent v/v) (per cent v/v)
Test solution. Dissolve 75 mg of the substance under 0 75 25
examination in 25 ml of acetonitrile and dilute to 50 ml with 32 40 60
water.
34 40 60
Reference solution (a). Dissolve 7.5 mg of clarithromycin
36 75 25
RS in 2.5 ml of acetonitrile and dilute to 5.0 ml with water.
42 75 25
100 ml with mobile phase. Inject the reference solution. The test is not valid unless the
than 2.0 per cent.
Inject reference solution (b). Test is not valid unless the tailing
chromatogram obtained with the test solution, the area of any Storage. Store protected from moisture.
326
IP 2007 CLOBAZAM
Clarithromycin Tablets Inject the reference solution. The test is not valid unless the
tailing factor is not more than 2.0, the column efficiency in not
Clarithromycin Tablets contain not less than 90.0 per cent and less than 750 theoretical plates and the relative standard
not more than 110.0 per cent of the stated amount of deviation for replicate injections is not more than 2.0 per cent.
clarithromycin, C38H69NO13.
Identification Calculate the content of C38H69NO13.
In the Assay, the principal peak in the chromatogram obtained Storage. Store protected from moisture.
Tests Clobazam
Dissolution (2.5.2). H3C
O
Apparatus. No. 1 N
Medium. 900 ml of 0.1 M sodium acetate buffer.
Speed and time. 50 rpm and 30 minutes. Cl N
Withdraw a suitable volume of the medium and filter. O

Test solution. Use the filtrate.
Reference solution. Weigh accurately a suitable quantity of C16H13ClN2O2 Mol. Wt. 300.7
clarithromycin RS, dissolve in methanol, dilute with Clobazam is 7-chloro-1-methyl-5-phenyl-1,5-dihydro-3H-1,5-
dissolution medium to obtain a solution having a known benzodiazepine-2,4-dione.
concentration of about 0.125 mg per ml.
Clobazam contains not less than 97.0 per cent and not more
Chromatographic system as described under Assay. than 103.0 per cent of C16H13ClN2O2, calculated on the dried
Calculate the content of C38H69NO13 in the tablet. basis.
D. Not less than 75 per cent of the stated amount of Description. A white or almost white, crystalline powder.
C38H69NO13.
Identification
on 1 g by drying in an oven at 110º, under vacuum, for 3 hours. Determine by infrared absorption spectrophotometry (2.4.6).
Assay. Determine by liquid chromatography (2.4.14). Compare the spectrum with that obtained with clobazam RS
or with the reference spectrum of clobazam.
Test solution. Disperse a quantity of powdered tablet
containing 125 mg of Clarithromycin in 200.0 ml with methanol Tests
and filter. Dilute 5.0 ml of the solution to 25.0 ml with mobile
phase. Related substances. Determine by liquid chromatography
(2.4.14).
clarithromycin RS in methanol. Dilute 5.0 ml of the solution Test solution. Dissolve 10 mg of the substance under
to 25.0 ml with mobile phase. examination in the mobile phase and dilute to 50 ml with the
mobile phase.
– a stainless steel column 15 cm x 4.6 mm packed with Reference solution (a). Dissolve 5.0 mg of 7-chloro-5-phenyl-
octadecylsilane bonded to porous silica (5 µm), 1,5-dihydro-3H-1,5-benzodiazepine-2,4-dione RS ( clobazam
– column temperature 50º, impurity A) in the mobile phase and dilute to 50 ml with the
– mobile phase: a mixture of 65 volumes of methanol and mobile phase. Dilute 1 ml of this solution to 100 ml with the
35 volumes of 0.067 M monobasic potassium phosphate mobile phase.
adjusted the pH to 4.0 with orthophosphoric acid, Reference solution (b). Dissolve 5 mg of chlordiazepoxide
– flow rate. 1 ml per minute, RS and 5 mg of clobazam RS in the mobile phase and dilute to
– spectrophotometer set at 210 nm, 50 ml with the mobile phase. Dilute 1 ml of the solution to 100
– a 50 µl loop injector. ml with the mobile phase.
327
CLOBAZAM CAPSULES IP 2007
Reference solution (c). Dilute 1 ml of the test solution to 200 to dryness. Dissolve the residue in the minimum amount of
ml with the mobile phase. methanol, evaporate to dryness and dry the residue at 105°
Chromatographic system for 10 minutes. The residue complies with the following test.
– a stainless steel column 25 cm x 4.6 mm, packed with Determine by infrared absorption spectrophotometry (2.4.6).
octadecylsilane bonded to porous silica (5 µm), Compare the spectrum with that obtained with the spectrum
– mobile phase: a mixture of 40 volumes of acetonitrile obtained with clobazam RS treated in the same manner or
and 60 volumes of water, with the reference spectrum of clobazam.
Inject reference solution (b). The resolution between the peaks (2.4.14).
due to chlordiazepoxide and clobazam is not less than 1.3.
Test solution. Extract a quantity of the contents of the capsules
Inject the test solution and reference solutions (a) and (c). containing 40 mg of Clobazam with three quantities, each of
Continue the chromatography for 5 times the retention time of 10 ml, of dichloromethane, combine the filtered extracts,
clobazam (about 15 minutes). In the chromatogram obtained evaporate to dryness and dissolve the residue in 2 ml of
with the test solution the area of the peak obtained due to methanol.
impurity A is not more than the area of the principal peak in the Reference solution (a). Dilute 1 volume of the test solution to
chromatogram obtained with reference solution (a) (0.5 per 200 volumes with methanol.
cent). The area of any other impurity peak is not more than 0.4
times the area of the principal peak in the chromatogram Reference solution (b). Dilute 1 volume of reference solution
obtained with reference solution (c) (0.2 per cent). The sum of (a) to 2.5 volumes with methanol.
the areas of all other impurity peaks is not more than twice the Chromatographic system
area of the principal peak in the chromatogram obtained with – a stainless steel column 25 cm x 4.6 mm, packed with
reference solution (c) (1.0 per cent). Ignore any peak with an octadecylsilane bonded to porous silica (5 µm),
area less than 0.1 times the area of the principal peak in the – mobile phase: a mixture of 40 volumes of acetonitrile
chromatogram obtained with reference solution (c) (0.05 per and 60 volumes of water,
cent). – flow rate. 1 ml per minute,
Sulphated ash (2.3.18). Not more than 0.1 per cent, determined – spectrophotometer set at 230 nm,
on the residue obtained in the test for Loss on drying. – a 20 µl loop injector.
Inject the test solution and reference solutions (a) and (b).
Continue the chromatography for 5 times the retention time of
on 1.0 g by drying in an oven at 100º-105º.
clobazam (about 15 minutes). In the chromatogram obtained
Assay. Weigh accurately about 50 mg and dissolve in 100.0 ml with the test solution the area of any impurity peak is not more
of ethanol (95 per cent). Dilute 2.0 ml of the solution to 250.0 than the area of the principal peak in the chromatogram
ml with the same solvent. Measure the absorbance of the obtained with reference solution (b). The sum of the areas of
resulting solution at the maximum at about 232 nm (2.4.7), all the impurity peaks is not more than twice the area of the
taking 1380 as the specific absorbance at 232 nm. principal peak in the chromatogram obtained with reference
Calculate the content of C16H13ClN2O2. solution (a).
Storage. Store protected from moisture. Dissolution (2.5.2).

Apparatus. No 1
Clobazam Capsules
Clobazam Capsules contain not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of clobazam, Determine by liquid chromatography (2.4.14).
C16H13ClN2O2. Test solution. The filtrate obtained as given above.
Identification Reference solution. Weigh accurately a suitable quantity of

clobazam RS, dissolve in methanol, and dilute with the
Shake a quantity of the capsules containing 20 mg of Clobazam dissolution medium to obtain a solution having the same
with 10 ml of dichloromethane, filter and evaporate the filtrate concentration as that of the test solution.
328
IP 2007 CLOFAZIMINE
Chromatographic system Calculate the content of C16H13ClN2O2 in the capsules.

– a stainless steel column 12.5 cm x 4.0 mm, packed with
octadecylsilane bonded to porous silica (5 ì m)(such as
Superspher 100RP-18),
– mobile phase: a mixture of 470 volumes of acetonitrile Clofazimine
– flow rate. 0.7 ml per minute, Cl
Inject alternatively the test solution and the reference solution. CH3
Calculate the content of C16H13ClN2O2 in the medium. N N CH3
C16H13ClN2O2. N N Cl
H
Assay. Determine by liquid chromatography (2.4.14). C27H22Cl2N4 Mol. Wt. 473.4
Test solution. Weigh 20 capsules. Open the capsules without Clofazimine is 3-(4-chloroanilino)-10-(4-chlorophenyl)- 2,10-
losing any part of the shells and transfer the contents as dihydro-2-(isopropylimino)phenazine.
completely as possible to a flask. Wash the shells with three
quantities, each of 30 ml, of methanol, add the washings to Clofazimine contains not less than 98.5 per cent and not more
the flask and dilute to 200.0 ml with methanol. Allow the shells than 101.5 per cent of C27H22Cl2N4, calculated on the dried
to dry at room temperature and weigh. The difference between basis.
the weights represents the weight of the total contents. Mix Description. Dark red crystals or a reddish-brown, fine powder;
the contents of the flask with the aid of ultrasound for 10 almost odourless.
minutes and stir magnetically for 20 minutes. Centrifuge a
portion of the suspension and dilute a volume of the resulting Identification
supernatant liquid containing 5 mg of Clobazam to 100.0 ml
with methanol.
Compare the spectrum with that obtained with clofazimine RS
Reference solution (a). A 0.005 per cent w/v solution of or with the reference spectrum of clofazimine.
clobazam RS in methanol.
B. When examined in the range 230 nm to 600 nm, a 0.0005 per
Reference solution (b). A solution containing 0.006 per cent cent w/v solution in 0.01 M methanolic hydrochloric acid
w/v of 7-chloro-1,5-dihydro-5-phenyl-1,5-benzodiazepine- shows absorption maxima, at about 283 nm and 487 nm;
2,4(3H)-dione RS (desmethylclobazam) and 0.0125 per cent absorbance at about 283 nm, about 0.65 and at about 487 nm,
w/v of clobazam RS in methanol. about 0.32 (2.4.7).
Chromatographic system C. Dissolve 2 mg in 3 ml of acetone and add 0.1 ml of
– a stainless steel column 20 cm x 4.6 mm, packed with hydrochloric acid; an intense violet colour is produced. Add
octadecylsilane bonded to porous silica (5 µm), 0.5 ml of 5 M sodium hydroxide; the colour changes to orange-
– mobile phase: a mixture of 470 volumes of acetonitrile red.
– flow rate 0.7 ml per minute, Tests
(2.4.17), coating the plate with silica gel F254 and exposing
Inject reference solution (b). The resolution between the peaks the plate to ammonia vapour immediately before use by
corresponding to desmethylclobazam and clobazam is not less suspending the plate for 30 minutes in a tank containing a
than 3. shallow layer of 0.2 M ammonia.
Inject reference solution (a). The test is not valid unless the Mobile phase. A mixture of 85 volumes of dichloromethane
relative standard deviation for replicate injections is not more and 4 volumes of 1-propanol.
than 2.0 per cent.
Inject alternatively the test solution and reference solution (a). examination in 10 ml of choroform.
329
CLOFAZIMINE CAPSULES IP 2007
Reference solution (a). A 0.016 per cent w/v solution of the chloroform to produce 50.0 ml. Measure the absorbance of
substance under examination in chloroform. the resulting solution at the maximum at about 491 nm (2.4.7),
Reference solution (b). A 0.01 per cent w/v solution of the using as the blank a mixture of 5.0 ml of 0.1 M methanolic
substance under examination in chloroform. hydrochloric acid and sufficient chloroform to produce
50.0 ml. Calculate the content of C27H22Cl2N4. taking 650 as the
Apply to the plate 5 µl of each solution. Allow the mobile specific absorbance at 491 nm.
phase to rise 12 cm. Dry the plate in air for 5 minutes and
replace it in the tank. When the mobile phase has again risen
12 cm dry the plate in air for 5 minutes and examine in daylight
and then in ultraviolet light at 254 nm. Spray the plate with
sulphuric acid (50 per cent) and examine again in daylight. Clomifene Citrate
test solution is not more intense than the spot in the Clomiphene Citrate
more than two such spots are more intense than the spot in O
the chromatogram obtained with reference solution (b). N CH3
HO COOH
Heavy metals (2.3.13). 2.0 g complies with the limit test for CH3 , HOOC COOH
Cl
on 1.0 g by drying in an oven at 105°. C26H28ClNO,C6H8O7 Mol . Wt. 598.1
Assay. Weigh accurately about 0.5 g and dissolve in 20 ml of Clomifene citrate is a mixture of E- and Z-isomers of 2-[4-(2-
chloroform. Add 50 ml of acetone and titrate with 0.1 M chloro-1,2-diphenylvinyl)phenoxy]triethylamine dihydrogen
perchloric acid in dioxan, determining the end-point citrate.
Clomifene Citrate contains not less than 98.0 per cent and not
1 ml of 0.1 M perchloric acid is equivalent to 0.04734 g of more than 101.0 per cent of C26H28ClNO,C6H8O7, calculated on
C27H22Cl2N4. the anhydrous basis.
Description. A white or pale yellow, crystalline powder.
Identification
Clofazimine Capsules
Tests B and C may be omitted if tests A and D are carried out.
Clofazimine Capsules contain not less than 95.0 per cent and Test A may be omitted if Tests B, C and D are carried out.
clofazimine, C27H22Cl2N4. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with clomifene citrate
Identification RS.
To 5 mg of the contents of a capsule add 3 ml of chloroform B. Dissolve about 5 mg in 5 ml of a mixture of 10 volumes of
and 1 ml of 2 M hydrochloric acid; the colour of the chloroform acetic anhydride and 50 volumes of pyridine and heat in a
layer changes to violet. Add 2 ml of 2 M sodium hydroxide; water-bath; a deep red colour is produced.
the colour changes to brownish-yellow. C. In the test for Related substances, the principal peak in the
Tests the peak in the chromatogram obtained with the reference
Other tests. Comply with the tests stated under Capsules. solution (a).
Assay. Weigh accurately a quantity of the mixed contents of D. A 5 per cent w/v solution gives the reactions of citrates
20 capsules containing about 0.15 g of Clofazimine and dissolve (2.3.1).
in sufficient chloroform to produce 100.0 ml. Filter through a
Tests
chloroform-washed plug of cotton wool. Dilute 5.0 ml of the
clear filtrate to 100.0 ml with chloroform. To 5.0 ml add 5.0 ml of NOTE — In the following tests, the solutions should be
0.1 M methanolic hydrochloric acid and sufficient protected from light in amber-coloured glassware. Ensure
330
IP 2007 CLOMIFENE CITRATE
minimum exposure of the solutions to daylight until they are chromatogram obtained with reference solution (b) (0.05 per
required for chromatography. cent).
Related substances. Determine by liquid chromatography Z-isomer. 30 to 50 per cent.
(2.4.14)
under examination in the mobile phase.
examination in 25 ml of 0.1 M hydrochloric acid, add 5 ml of
Reference solution (a). A solution containing 0.125 per cent 1 M sodium hydroxide and shake with three quantities, each
w/v of clomifene citrate for performance test RS in the mobile of 25 ml, of ethanol-free chloroform. Wash the combined
phase. extracts with 10 ml of water, dry over anhydrous sodium
sulphate and dilute to 100.0 ml with ethanol-free chloroform
To 20.0 ml of the solution add 0.1 ml of triethylamine and
dilute to 100.0 ml with hexane.
Reference solution. Dissolve 25 mg of clomifene citrate RS in
25 ml of 0.1 M hydrochloric acid, add 5 ml of 1 M sodium
butylsilyl silica gel (such as Vydac C4),
hydroxide and shake with three quantities, each of 25 ml, of
– mobile phase: mix 400 volumes of acetonitrile with
ethanol-free chloroform. Wash the combined extracts with
600 volumes of water and add 8 ml of diethylamine,
10 ml of water, dry over anhydrous sodium sulphate and dilute
adjust the pH of the mixture to 6.2 by the addition of
to 100.0 ml with ethanol-free chloroform. To 20.0 ml of the
about 1 to 2 ml of phosphoric acid taking care to reduce
solution add 0.1 ml of triethylamine and dilute to 100.0 ml with
progressively the volume of each addition as the required
hexane.
pH is approached,
– flow rate. 1.2 ml per minute, Chromatographic system
– spectrophotometer set at 233 nm, – a stainless steel column 30 cm x 4 mm, packed with porous
– a 10 µl loop injector. silica particles (10 µm) (such as Parasol),
– mobile phase: a mixture of 1 volume of triethylamine,
Equilibrate the column with the mobile phase at a flow rate of 200 volumes of ethanol-free chloroform and
1.2 ml per minute for about one hour. 800 volumes of hexane,
Inject reference solution (a). Continue the chromatography – flow rate. 2 ml per minute,
for twice the retention time of the principal peak. Measure the – spectrophotometer set at 302 nm,
height (a) above the baseline of the peak due to clomifene – a 50 µl loop injector.
impurity A and the height (B) above the baseline of the lowest Equilibrate the column with the mobile phase for about 2 hours.
point of the curve separating this peak from the peak due to
clomifene. The test is not valid unless A is greater than Inject the reference solution. The chromatogram obtained
15 times B and the chromatogram obtained resembles the shows a peak due to E-isomer just before a peak due to
reference chromatogram. If necessary, adjust the concentration Z-isomer. The test is not valid unless the resolution between
of acetonitrile in the mobile phase. the peaks corresponding to E- and Z-isomers is at least 1.0. If
necessary, adjust the relative proportions of ethanol-free
Inject separately the test solution and reference solution (b). chloroform and hexane in the mobile phase. Measure the area
Continue the chromatography for four times the retention time of the peak due to the Z-isomer in the chromatogram obtained
of the principal peak. In the chromatogram obtained with the with the test solution and the reference solution.
test solution the area of any peak due to 2-[4-(1,2-
diphenylvinyl)phenoxy]triethylamine is not greater than that Calculate the content of Z-isomer as a percentage of the total
of the principal peak in the chromatogram obtained with clomifene citrate present.
reference solution (b) (2 per cent) and the area of any other Water (2.3.43). Not more than 1.0 per cent, determined on
secondary peak is not greater than half the area of the principal 1.0 g.
(b) (1 per cent); the sum of the areas of any secondary peaks Assay. Weigh accurately about 0.5 g and dissolve in 50 ml of
is not greater than 1.25 times the area of the principal peak in anhydrous acetic acid. Titrate with 0.1 M perchloric acid,
the chromatogram obtained with reference solution (b) determining the end-point potentiometrically (2.4.25). Carry
(2.5 per cent). Ignore any peak with a retention time relative to out a blank titration.
the clomifene peak of 0.2 or less and any peak with an area 1 ml of 0.1 M perchloric acid is equivalent to 0.05981 g of
less than 0.025 times the area of the principal peak in the C26H28ClNO,C6H8O7.
331
CLOMIFENE TABLETS IP 2007
Clomifene Tablets Calculate the percentage of Z-isomer from the expression

100Az/(1.08AE+AZ) where AZ and AE are the areas of the peaks
Clomifene Citrate Tablets; Clomiphene Tablets; due to the Z- and E-isomers respectively.
Clomophene Tablets Dissolution (2.5.2).
Clomifene Tablets contain not less than 92.5 per cent and not
Apparatus. No 1
more than 207.5 per cent of the stated amount of clomifene
Medium. 900 ml of water
citrate, C26H28ClNO,C6H8O7.
Identification Withdraw a suitable volume of the medium and filter promptly
A. When examined in the range 230 nm to 360 nm (2.4.7), the through a membrane filter disc having an average pore diameter
solution obtained in the Assay shows absorption maxima at not greater than 1.0 µm, rejecting the first 1 ml of the filtrate.
about 235 nm and 292 nm. Dilute a suitable volume of the filtrate with 0.1 M hydrochloric
acid. Measure the absorbance of the resulting solution at the
B. Dissolve a quantity of the powdered tablets containing
5 mg of Clomifene Citrate in 5 ml of a mixture of 10 volumes of
C26H28ClNO,C6H8O7 in the medium taking 317 as the specific
acetic anhydride and 50 volumes of pyridine and heat in a
water-bath; a deep red colour is produced.
Tests C26H28ClNO,C6H8O7.
Z-isomer. 30 to 50 per cent of the content of clomifene citrate Assay. Weigh and powder 20 tablets. Weigh accurately a
as determined in the Assay. quantity of the powder containing about 50 mg of Clomifene
Determine by liquid chromatography (2.4.14) Citrate, shake for 30 minutes with 70 ml of 0.1 M hydrochloric
Test solution. Shake a quantity of the powdered tablets acid prepared in a 30 per cent v/v solution of 2-propanol
containing about 50 mg of Clomifene Citrate with 50 ml of (instead of water normally used for the purpose as solvent),
0.1 M hydrochloric acid for 10 minutes and filter. To 25 ml of dilute to 100.0 ml with the propanolic hydrochloric acid and
the filtrate add 5 ml of 1 M sodium hydroxide and extract with filter. Dissolve 5.0 ml of the filtrate to 100.0 ml with 0.1 M
three quantities, each of 25 ml, of ethanol-free chloroform. hydrochloric acid and measure the absorbance of the resulting
Wash the combined extracts with 10 ml of water, dry over solution at the maximum at about 292 nm (2.4.7), using a
anhydrous sodium sulphate and add sufficient ethanol-free solution prepared by diluting 5 ml of the propanolic
chloroform to produce 100.0 ml. To 20.0 ml of the resulting hydrochloric acid to 100 ml with 0.1 M hydrochloric acid as
solution add 0.1 ml of triethylamine and sufficient hexane to the blank. Calculate the content of C26H28ClNO,C6H8O7 taking
produce 100 ml. 175 as the specific absorbance at 292 nm.
– a stainless steel column 30 cm x 4 mm, packed with porous
silica particles (10 µm) (such as Parasol), Clomipramine Hydrochloride
– mobile phase: a mixture of ethanol-free chloroform and
hexane, each containing 0.10 per cent v/v of
Cl
triethylamine, adjusted so that the baseline separation
is obtained between E- and Z-isomers of clomifene (a
mixture of 20 volumes of ethanol-free chloroform and
80 volumes of hexane is suitable), CH3
N N , HCl
– spectrophotometer set at 302 nm, CH3
Stabilise the system by passing about 250 ml of the mobile
phase. C19H23ClN2,HCl Mol. Wt. 351.3
Inject the test solution. In the chromatogram a peak due to the
Clomipramine is 3-(3-chloro-10,11-dihydro-5H-dibenzo[b,f]
E-isomer precedes that due to the Z-isomer of clomifene. The
azepin-5-yl)-N,N-dimethylpropan-1-amine hydrochloride.
test is not valid unless baseline separation is achieved between
E- and Z-clomifene and the column efficiency is greater than Clomipramine Hydrochloride contains not less than 99.0 per
10,000 theoretical plates per metre determine using the peak cent and not more than 101.0 per cent of C19H23ClN2,HCl,
due to E-isomer. calculated on the dried basis.
332
IP 2007 CLOMIPRAMINE CAPSULES
Description. A white or slightly yellow, crystalline powder, – mobile phase: A. 1.2 g of sodium dihydrogen phosphate
slightly hygroscopic. in water, add 1.1 ml of nonylamine, adjust to pH 3.0 with
phosphoric acid and dilute to 1000 ml with water,
Identification B. acetonitrile,
Test A may be omitted if tests B, C and D are carried out. Tests – a linear gradient programme using the conditions given
B, C and D may be omitted if test A is carried out. below,
A. Determine by infrared absorption spectrophotometry (2.4.6). – spectrophotometer set at 254 nm,
Compare the spectrum with that obtained with clomipramine – a 20 µl loop injector.
hydrochloride RS.
B. Determine by thin-layer chromatography (2.4.17), coating (in min.) (per cent v/v) (per cent v/v)
0 75 25
Mobile phase. A mixture of 5 volumes of ammonia, 25 volumes 10 65 35
of acetone and 75 volumes of ethyl acetate. 32 65 35
Test solution. Dissolve 20 mg of the substance under 44 75 25
examination in 10 ml of methanol. Inject reference solution (a). Test is not valid unless the column
Reference solution. A 0.2 per cent w/v solution of effciency is not less than 2000 theoretical plates and the tailing
clomipramine hydrochloride RS in methanol. factor is not more than 2.0.
Apply to the plate 5 µl of each solution. Allow the mobile Inject the test solution and reference solution (b). In the
phase to rise 15 cm. Dry the plate in air, spray with 0.5 per cent chromatogram obtained with the test solution, the area of any
solution of potassium dichromate in a 20 per cent solution of secondary peak is not more than 0.5 times the area of the peak
sulphuric acid. The principal spot in the chromatogram in the chromatogram obtained with reference solution (b)
obtained with the test solution corresponds to the principal (0.5 per cent) and the sum of areas of all the secondary peaks
spot in the chromatogram obtained with the reference solution. is not more than the area of the peak in the chromatogram
C. Dissolve about 5 mg in 2 ml of nitric acid. An intense blue obtained with the reference solution (b) (1.0 per cent).
colour develops. Heavy metals (2.3.13). 2 g complies with limit test for heavy
D. Dissolve about 50 mg in 5 ml of water and add 1 ml of dilute
ammonia. Allow to stand for 5 minutes and filter. Acidify the Sulphated ash (2.3.18). Not more than 0.1 per cent.
filtrate with dilute nitric acid. The solution gives reaction A Loss on drying (2.4.19). Not more than 0.5 per cent, determined
of chlorides (2.3.1). on 1 g by drying in an oven at 105º.
Tests Assay. Weigh accurately about 0.25 g, dissolve in 50.0 ml of
ethanol and add 5.0 ml of 0.01 M hydrochloric acid. Titrate
Appearance of solution. A 10 per cent w/v solution in carbon with 0.1 M sodium hydroxide. Determine the end-point
dioxide-free water (Solution A) is clear (2.4.1) and not more potentiometrically (2.4.25).
intensely coloured than reference solution Y5 (2.4.1). 1 ml of 0.1 M sodium hydroxide is equivalent to 0.03513 g of
pH (2.4.24). 3.5 to 5.0, determined in solution A. C19H24Cl2N2.
Related substances. Determine by liquid chromatography Storage. Store protected from light and moisture.
(2.4.14).
Solvent mixture. 25 volumes of mobile phase B and 75 volumes
of mobile phase A. Clomipramine Capsules
Test solution. Dissolve 20 mg of the substance under Clomipramine Hydrochloride Capsules
examination in 10 ml of solvent mixture.
Clomipramine Capsules contain Clomipramine Hydrochloride.
Clomipramine Capsules contains not less than 95.0 per cent
clomipramine hydrochloride RS in solvent mixture.
Reference solution (b). Dilute 1 ml of reference solution (a) to clomipramine hydrochloride, C19H23ClN2,HCl.
100 ml with solvent mixture.
– a stainless steel column 25 cm x 4.6 mm packed with Triturate a quantity of the contents of the capsules containing
cyanopropylsilyl silica (5 µm), 0.15 g of Clomipramine Hydrochloride with 10 ml of chloroform,
333
CLONAZEPAM IP 2007
filter and evaporate the filtrate to dryness. The residue complies Assay. Determine by liquid chromatography (2.4.14).
with the following test. Test solution. Weigh accurately a quantity of the mixed
Determine by infrared absorption spectrophotometry (2.4.6 ). contents of 20 capsules containing about 50 mg of
Compare the spectrum with that obtained with clomipramine Clomipramine, disperse in 100.0 ml of methanol. Dilute 5.0 ml
hydrochloride RS. of the solution to 50.0 ml with methanol.
Tests Reference solution. A 0.0125 per cent w/v solution of clobazam
RS in methanol.
(2.4.14). Chromatographic system
– a stainless steel column 20 cm × 4.6 mm packed with
Test solution. Disperse a quantity of the mixed contents of
20 capsules containing 20 mg of Clomipramine Hydrochloride
with 5 ml of mobile phase A with the aid of ultrasound for
15 minutes, dilute to 10 ml with the same solvent and filter.
– flow rate. 0.7 ml per minute.
Reference solution (a). A 0.2 per cent w/v solution of – spectrophotometer set at 230 nm,
clomipramine hydrochloride RS in mobile phase A. – a 20 µl loop injector.
Reference solution (b). Dilute 1 ml of reference solution (a) to Inject the reference solution. The test is not valid unless the
100 ml with mobile phase A. relative standard deviation for replicate injections is not more
Chromatographic system than 2.0 per cent.
– a stainless steel column 25 cm x 4.6 mm packed with Inject the test solution and the reference solution.
cyanopropylsilyl silic a (5 µm) (such as Hypersil BDS
CN), Calculate the content of C16H13ClN2O2.
– mobile phase: A. a mixture of 75 volumes of solution
prepared by dissolving 1.2 g of sodium dihydrogen
orthophosphate in 950 ml of water, add 1.1 ml of
nonylamine, adjust to pH 3.0 with orthophosphoric acid
Clonazepam
and add sufficient water to produce 1000 ml (solution
A) and 25 volumes of acetonitrile. H O
B. a mixture of 65 volumes of solution A N
and 35 volumes of acetonitrile,
– a linear gradient programme using the conditions given O2 N N
below,
– flow rate. 1.5 ml per minute, Cl
Time Mobile phase A Mobile phase B C15H10ClN3O3 Mol. Wt. 315.7
Clonazepam is 5-(2-chlorophenyl)-7-nitro-1,3-dihydro-2H-
0 75 25
1,4-benzodiazepin-2-one.
10 65 35
Clonazepam contains not less than 99.0 per cent and not more
32 65 35
than 101.0 per cent of C15H10ClN3O3, calculated on the dried
44 75 25 basis.
Inject reference solution (a). Test is not valid unless the column Description. A slightly yellowish, crystalline powder.
effciency is not less than 2000 theoretical plates and the tailing
factor is not more than 2.0. Identification
Compare the spectrum with that obtained with clonazepam RS.
in the chromatogram obtained with reference solution (b) Tests
is not more than the area of the peak in the chromatogram Related substances. Determine by liquid chromatography
obtained with the reference solution (b) (1.0 per cent). (2.4.14).
334
IP 2007 CLONAZEPAM INJECTION
Solvent mixture. 10 volumes of tetrahydrofuran, 42 volumes The injection is constituted by dissolving the contents of the
of methanol and 48 volumes of water. sealed container in the requisite amount of sterile Water for
Test solution. Dissolve 0.05 g of the substance under Injections, immediately before use.
examination in 10 ml of methanol and dilute to 100 ml with the The constituted solution complies with the requirements for
solvent mixture. Clarity of solution and Particulate matter stated under
Reference solution (a). Dissolve 25 mg of clonazepam RS in
5 ml of methanol and dilute to 50 ml with the solvent mixture. Storage. The constituted solution should be used immediately
Clonazepam Injection contains not less than 95.0 per cent and
clonazepam, C15H10ClN3O3.
octylsilyl silica (5 µm),
– mobile phase: a mixture of 10 volumes of tetra- Description. A slightly yellowish, crystalline powder.
hydrofuran, 42 volumes of methanol and The contents of the sealed container comply with the
48 volumes of a 0.66 per cent solution of ammonium requirements stated under Parenteral Preparations
phosphate previously adjusted to pH 8.0 with a 4 per (Powders for Injection) and with the following requirements.
cent w/v of sodium hydroxide or orthophosphoric
acid, Identification
plate with silica gel GF254.
Mobile phase. A mixture of 2 volumes of strong ammonia
solution, 15 volumes of n-heptane, 30 volumes of
nitromethane and 60 volumes of ether.
Test solution. Dilute 3 ml of a solution containing 3 mg of
Inject the test solution and reference solution (b). In the Clonazepam in a stoppered tube with an equal volume of water,
chromatogram obtained with the test solution, the area of any shake with 1 ml of chloroform, allow to separate and use the
secondary peak is not more than 0.5 times the area of the peak chloroform layer.
(0.5 per cent) and the sum of areas of all the secondary peaks Reference solution. Dissolve 3 mg of clonazepam RS in 1 ml
is not more than the area of the peak in the chromatogram of chloroform.
obtained with the reference solution (b) (1.0 per cent). Apply to the plate 10 µl of each solution. Allow the mobile
Sulphated ash (2.3.18). Not more than 0.1 per cent. phase to rise 10 cm. After development, dry the plate in a
current of cold air, spray with 2 M sodium hydroxide and heat
Loss on drying (2.4.19). Not more than 0.5 per cent, determined at 120° for 15 minutes. The yellow spot in the chromatogram
on 1 g by drying in an oven at 105º for 4 hours. obtained with test solution corresponds to that in the
Assay. Weigh accurately about 0.275 g, dissolve in 50 ml of chromatogram obtained with reference solution.
acetic anhydride. Titrate with 0.1 M perchloric acid.
Tests
Determine the end-point potentiometrically (2.4.25). Carry out
a blank titration. pH (2.4.24). 3.4 to 4.3.
1 ml of 0.1 M perchloric acid is equivalent to 0.03157g of Related substances. Determine by thin-layer chromatography
C15H10ClN3O3. (2.4.17), coating the plate with silica gel G.
Storage. Store protected from light. Mobile phase. A mixture of 20 volumes of chloroform and 80
volumes of ether.
Test solution. Dilute, if necessary, a volume of the solution
containing 10 mg of Clonazepam to 20 ml with water and extract
Clonazepam Injection with three 3 ml quantities of chloroform. Wash each chloroform
Clonazepam Injection is a sterile material consisting of extract separately with the same 10 ml volume of water,
Clonazepam with or without excipients. It is filled in a sealed combine the extracts and add sufficient chloroform to produce
container. 10 ml.
335
CLONIDINE HYDROCHLORIDE IP 2007
Reference solution (a). A 0.0005 per cent w/v solution of 2- Clonidine Hydrochloride contains not less than 98.5 per cent
amino-2’-chloro-5-nitrobenzophenone RS and not more than 101.0 per cent of C9H9Cl2N3,HCl, calculated
(‘nitrobenzophenone’) in chloroform. on the dried basis.
Reference solution (b). A 0.0002 per cent w/v solution of 2- Description. A white or almost white, crystalline powder.
amino-2’-chloro-5-nitrobenzophenone RS
(‘nitrobenzophenone’) in chloroform. Identification
Reference solution (c). A 0.0002 per cent w/v of 3-amino-4- Test A may be omitted if tests B, C and D are carried out. Tests
(2-chlorophenyl)-6-nitroquinolin-2-one RS (‘carbostyril’) in B and C may be omitted if tests A and D are carried out.
chloroform.
Apply to the plate 50 µl of each solution. After development, Compare the spectrum with that obtained with clonidine
dry the plate in a current of cold air. For the second hydrochloride RS or with the reference spectrum of clonidine
development use a mixture of 10 volumes of ether and 90 hydrochloride.
volumes of nitromethane. After development, dry the plate,
heat at a pressure of 2kPa at 120° for 3 hours, allow to cool and B. When examined in the range 230 nm to 600 nm, a 0.03 per
spray with a 10 per cent w/v solution of zinc chloride in 0.1 M cent w/v solution in 0.01 M hydrochloric acid shows
hydrochloric acid. Dry the plate in air and examine in visual absorption maxima, at about 272 nm and 279 nm and an
light. Any spots in the chromatogram obtained with test inflection at about 265 nm (2.4.7)
solution corresponding to the nitrobenzophenone and C. In the test for Related substances, the principal spot in the
carbostyril impurities are not more intense than the spots in chromatogram obtained with test solution (b) corresponds to
the chromatograms obtained with reference solutions (b) and that in the chromatogram obtained with reference solution
(c) respectively (0.2 per cent). Any other secondary spot in (b).
the chromatogram obtained with test solution is not more
D. Gives reaction A of chlorides (2.3.1).
Tests
Assay. Protect the solutions from light throughout the assay.
To a volume of the solution containing 20 mg of Clonazepam, dioxide-free water is clear (2.4.1), and not more intensely
dilute to 100.0 ml with propan-2-ol. Dilute 10.0 ml of the coloured than reference solution YS7 (2.4.1).
solution to 100.0 ml with propan-2-ol . Measure the
absorbance of the resulting solution at the maximum at 310 nm pH (2.4.24). 4.0 to 5.0, determined in a 5.0 per cent w/v solution.
(2.4.7). Calculate the content of C15H10ClN3O3 taking 364 as Related substances. Determine by thin-layer chromatography
the specific absorbance at 310 nm. (2.4.17), coating the plate with silica gel G.
Storage. Store protected from light. Mobile phase. Shake together 50 volumes of water, 40 volumes
Labelling. The label states (1) ‘Sterile Clonazepam of 1-butanol and 10 volumes of glacial acetic acid and allow
Concentrate’; (2) that the diluted injection is to be given by the layers to separate. Use the filtered upper layer.
intravenous injection. Test solution (a). Dissolve 0.1 g of the substance under
Test solution (b). Dilute 1.0 ml of test solution (a) to 10 ml with
methanol.
Clonidine Hydrochloride
Reference solution (a). Dilute 5 ml of test solution (b) to
H Cl
N N Reference solution (b). A 0.1 per cent w/v solution of
,HCl clonidine hydrochloride RS in methanol.
N
H Cl Apply to the plate 10 µl of each solution. After development,
dry the plate in air and spray with modified potassium
iodobismuthate solution. Allow to dry in air for 1 hour, spray
C9H9Cl2N3,HCl Mol. Wt. 266.6
again with the same reagent and immediately spray with a
Clonidine Hydrochloride is 2-[(2,6-dichlorophenyl)imino]- 5 per cent w/v solution of sodium nitrite. Any secondary spot
imidazolidine hydrochloride. in the chromatogram obtained with test solution (a) is not
336
IP 2007 CLONIDINE TABLETS
more intense than the spot in the chromatogram obtained again with the same reagent and immediately spray with a
with reference solution (a). 5 per cent w/v solution of sodium nitrite. Any secondary spot
Sulphated ash (2.3.18). Not more than 0.2 per cent w/w. in the chromatogram obtained with the test solution is not
Loss on drying (2.4.19). Not more than 0.5 per cent, determined with the reference solution.
Assay. Weigh accurately about 0.2 g and dissolve in 70 ml of Preparations (Injections).
ethanol (95 per cent). Titrate with 0.1 M ethanolic sodium
Assay. To an accurately measured volume of the injection
hydroxide, determining the end-point potentiometrically
containing 150 µg of Clonidine Hydrochloride add 25 ml of
(2.4.25).
citrophosphate buffer pH 7.6, 5 ml of water, and 1 ml of a
1 ml of 0.1 M ethanolic sodium hydroxide is equivalent to solution containing 0.15 per cent w/v of bromothymol blue
0.02666 g of C9H9Cl2N3,HCl. and 0.15 per cent w/v of anhydrous sodium carbonate. Add
30 ml of chloroform, shake for 1 minute and centrifuge. To
15.0 ml of the chloroform layer add 10 ml of boric acid solution
and measure the absorbance of the resulting solution at the
Clonidine Injection maximum at about 420 nm (2.4.7), using as the blank a solution
Clonidine Hydrochloride Injection prepared by diluting 10 ml of boric acid solution to 25.0 ml
with chloroform. Repeat the operation by adding to 5.0 ml of
Clonidine Injection is a sterile solution of Clonidine
a 0.003 per cent w/v solution of clonidine hydrochloride RS,
Hydrochloride in Water for Injections.
previously dried to constant weight at 105°, 20 ml of
Clonidine Injection contains not less than 90.0 per cent and citrophosphate buffer pH 7.6 and completing the procedure
not more than 110.0 per cent of the stated amount of clonidine described above beginning at the words “5 ml of water”.
hydrochloride, C9H9Cl2N3,HCl. Calculate the content of C9H9Cl2N3,HCl from the absorbance
obtained using clonidine hydrochloride RS in place of the
Identification substance under examination.
A. Dilute a volume containing 300 µg of Clonidine Storage. Store in single dose containers.
Hydrochloride to 5 ml with 0.01 M hydrochloric acid. When
examined in the range 230 nm to 360 nm, the resulting solution
shows absorption maxima at about 272 nm and 279 nm and an
inflection at about 265 nm (2.4.7).
Clonidine Tablets
B. To a volume containing 150 µg of Clonidine Hydrochloride
add 1 ml of a 10 per cent w/v solution of ammonium reineckate Clonidine Hydrochloride Tablets
and keep aside for 5 minutes; a pink precipitate is obtained. Clonidine Tablets contain not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of clonidine
Tests hydrochloride, C9H9Cl2N3,HCl.
pH (2.4.24). 4.0 to 7.0.
Identification
(2.4.17), coating the plate with silica gel G. To a quantity of the powdered tablets containing 500 µg of
Clonidine Hydrochloride add 30 ml of water and 5 ml of 1 M
Mobile phase. Shake together 50 volumes of water, 40 volumes sodium hydroxide. Swirl gently and extract with 20 ml of
of 1-butanol and 10 volumes of glacial acetic acid and allow chloroform. Remove the chloroform layer, dry with anhydrous
the layers to separate. Use the filtered upper layer. sodium sulphate, filter and evaporate the filtrate to dryness.
Test solution. Add 10 ml of methanol to a volume containing Dissolve the residue in 8 ml of 0.01 M hydrochloric acid. The
750 µg of Clonidine Hydrochloride, evaporate to dryness and resulting solution complies with the following tests.
dissolve the residue in 0.5 ml of methanol. A. When examined in the range 230 nm to 360 nm, it shows
Reference solution. Dilute 1 volume of the test solution to absorption maxima at about 272 nm and 279 nm and an inflection
100 volumes with methanol. at about 265 nm (2.4.7).
Apply to the plate 20 µl of each solution. After development, B. To 2 ml add 1 ml of a 10 per cent w/v solution of ammonium
dry the plate in air and spray with modified potassium reineckate and allow to stand for 5 minutes; a pink precipitate
iodobismuthate solution. Allow to dry in air for 1 hour, spray is produced.
337
CLOTRIMAZOLE IP 2007
Tests Clotrimazole
Tablets.
For tablets containing 300 µg or more of Clonidine Cl
Hydrochloride − To one tablet add 200 ml of citrophosphate
buffer pH 7.6, shake until disintegrated and dilute with C
citrophosphate buffer pH 7.6, if necessary, to give a solution N
containing about 0.0015 per cent w/v of Clonidine
Hydrochloride. To 5 ml of the supernatant liquid add 1 ml of a N
solution containing 0.15 per cent w/v of bromothymol blue C22H17CIN2 Mol. Wt 344.8
and 0.15 per cent w/v of anhydrous sodium carbonate. Add
10 ml of chloroform, shake for 1 minute and centrifuge. To 5 ml Clotrimazole is 1-[(2-chlorophenyl)diphenylmethyl]-1H-
of the supernatant liquid add 5 ml of boric acid solution and imidazole.
measure the absorbance of a 2-cm layer of the resulting solution Clotrimazole contains not less than 98.0 per cent and not more
at the maximum at about 420 nm (2.4.7), using as the blank a than 102.0 per cent of C22H17ClN2, calculated on the dried basis.
mixture of 5 ml of boric acid solution and 5 ml of chloroform.
Repeat the operation by using a solution prepared by diluting Description. A white or pale yellow, crystalline powder.
5 ml of a 0.03 per cent w/v solution of clonidine hydrochloride Identification
RS to 100 ml with citrophosphate buffer pH 7.6, transferring
5 ml to a separator and completing the procedure described A. Determine by infrared absorption spectrophotometry (2.4.6).
above beginning at the words “add 1 ml of a solution...”. Compare the spectrum with that obtained with clotrimazole
RS or with the reference spectrum of clotrimazole.
Calculate the content of C9H9Cl2N3,HCl in the tablet from the
absorbance obtained using clonidine hydrochloride RS in B. When examined in the range 230 nm to 360 nm, a 0.04 per
place of the tablet. cent w/v solution in a mixture of 1 volume of 0.1 M hydrochloric
acid and 9 volumes of methanol shows absorption maxima at
For tablets containing less than 300 µg of Clonidine about 262 and 265 nm; absorbance at about 262 nm, about
Hydrochloride - Use the same procedure but with a 0.9 and at about 265 nm, about 0.92 (2.4.7).
concentration of 0.001 per cent w/v or 0.0005 per cent w/v of
Clonidine Hydrochloride as appropriate and with Tests
corresponding smaller concentrations of clonidine
Appearance of solution. A 5.0 per cent w/v solution in ethanol
hydrochloride RS.
(95 per cent) is clear (2.4.1), and not more intensely coloured
Other tests. Comply with the tests stated under Tablets. than reference solution BYS6 (2.4.1).
Assay. Weigh and powder 20 tablets. To an accurately weighed 2-Chlorotritanol. Determine by liquid chromatography
quantity of the powder containing about 100 µg of Clonidine (2.4.14).
Hydrochloride add 25 ml of citrophosphate buffer pH 7.6 and
Solvent mixture. A mixture of 70 volumes of methanol and 30
shake for 15 minutes. Add 5 ml of water and 1 ml of a solution
volumes of 0.02 M phosphoric acid.
containing 0.15 per cent w/v of bromothymol blue and
0.15 per cent w/v of anhydrous sodium carbonate and shake Test solution. Dissolve 0.1 g of the substance under
to disperse. Add 30 ml of chloroform, shake for 1 minute and examination in 100 ml with solvent mixture.
centrifuge. To 15.0 ml of the chloroform layer add 10 ml of Reference solution (a). A 0.0002 per cent w/v solution of
boric acid solution and measure the absorbance of the 2-chlorotritanol RS in the same solvent mixture.
resulting solution at the maximum at about 420 nm (2.4.7),
using as the blank a solution prepared by diluting 10 ml of Reference solution (b). Dilute 1 volume of the test solution to
boric acid solution to 25.0 ml with chloroform. Repeat the 50 volumes with the same solvent mixture.
operation by adding to 5.0 ml of a 0.003 per cent w/v solution Chromatographic system
of clonidine hydrochloride RS, previously dried to constant – a stainless steel column 25 cm x 4.6 mm, packed with
weight at 105°, 20 ml of citrophosphate buffer pH 7.6 and octadecylsilyl silica gel (5 µm),
completing the procedure described above beginning at the – mobile phase: a mixture of 70 volumes of methanol and
words “5 ml of water”. Calculate the content of C9H9Cl2N3,HCl 30 volumes of 0.02 M phosphoric acid adjusted to
from the absorbance obtained using clonidine hydrochloride pH 7.5 with a 10 per cent w/v solution of triethylamine
RS in place of the substance under examination. in methanol,
338
IP 2007 CLOTRIMAZOLE CREAM
– flow rate. 1.5 ml per minute, Identification

– a 20 µl loop injector. A. Mix a quantity of the cream containing 40 mg of Clotrimazole
with 20 ml of a mixture of 1 volume of 1 M sulphuric acid and
Inject the test solution and reference solution (a). Allow the 4 volumes of methanol and shake with two quantities, each of
chromatography to proceed for 1.5 times the retention time of 50 ml, of carbon tetrachloride, discarding the organic layers.
the principal peak in the chromatogram obtained with the test Make the aqueous phase alkaline with dilute ammonia
solution. The area of any peak corresponding to solution, add a further 5 ml of dilute ammonia solution and
2-chlorotritanol in the chromatogram obtained with the test extract with two quantities, each of 40 ml, of chloroform.
solution is not greater then the area of the peak in the Combine the chloroform extracts, shake with 5 g of anhydrous
chromatogram obtained with reference solution (a). The test sodium sulphate, filter and add sufficient chloroform to the
is not valid unless the column efficiency, determined using filtrate to produce 100 ml. Evaporate 50 ml to dryness and
the principal peak in the chromatogram obtained with reference dissolve the residue in 50 ml of a mixture of 1 volume of 0.1 M
solution (b) is not less than 9000 theoretical plates. hydrochloric acid and 9 volumes of methanol. When examined
Imidazole. Determine by thin-layer chromatography (2.4.17), in the range 230 nm to 360 nm, the resulting solution shows
coating the plate with silica gel G. absorption maxima at about 262 nm and 265 nm (2.4.7).
Mobile phase. A mixture of 180 volumes of toluene, 20 volumes B. Determine by thin-layer chromatography (2.4.17), coating
of 1-propanol and 1 volume of strong ammonia solution. the plate with silica gel G.
Test solution. Dissolve 0.5 g of the substance under Mobile phase. Di-isopropyl ether in a chromatography tank,
examination in 10 ml of chloroform. containing 25 ml of strong ammonia solution in a beaker.
Reference solution. A 0.01 per cent w/v solution of imidazole Test solution. Shake a quantity of the cream containing 20 mg
in chloroform. of Clotrimazole with 4 ml of dichloromethane for 30 minutes,
centrifuge and use the supernatant liquid.
dry the plate in air, place in a tank of chlorine prepared by the Reference solution. A 0.5 per cent w/v solution of clotrimazole
addition of hydrochloric acid to potassium permanganate RS in dichloromethane.
and allow to stand for 2 minutes. Remove any excess chlorine Apply to the plate 10 µl of each solution. After development,
from the plate with a current of air and spray with potassium dry the plate in a current of air and spray with dilute potassium
iodide and starch solution. Any spot corresponding to iodobismuthate solution. The spot in the chromatogram
imidazole in the chromatogram obtained with the test solution obtained with the test solution is reddish brown and
is not more intense than the spot in the chromatogram obtained corresponds to the spot in the chromatogram obtained with
with the reference solution. the reference solution.
Tests
on 1.0 g by drying in an oven at 105°. 2-Chlorotritanol. Determine by liquid chromatography
(2.4.14).
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Test solution. Extract a quantity of the cream containing 20 mg
acid, using 1-naphtholbenzein solution as indicator to a green of Clotrimazole by warming with 20 ml of methanol in a water-
end-point. Carry out a blank titration. bath at 50° for 5 minutes, shaking occasionally. Remove from
the water-bath, shake the mixture vigorously while cooling to
1 ml of 0.1 M perchloric acid is equivalent to 0.03448 g of room temperature, cool in ice for 15 minutes, centrifuge for 5
C22H17ClN2. minutes and decant the supernatant liquid. Repeat the
Storage. Store protected from light. extraction with further quantities, each of 20 ml, of methanol.
To the combined methanol extracts add 10 ml of methanol and
dilute to 100.0 ml with 0.02 M phosphoric acid. Cool in ice
and filter through a membrane filter.
Clotrimazole Cream
Clotrimazole Cream contains Clotrimazole in a suitable base. 2-chlorotritanol RS in a mixture of 70 volumes of methanol
Clotrimazole Cream contains not less than 95.0 per cent and and 30 volumes of 0.02 M phosphoric acid.
not more than 105.0 per cent of the stated amount of Reference solution (b). Dilute 1 volume of the test solution to
clotrimazole, C22H17ClN2. 50 volumes with the same solvent mixture.
339
CLOTRIMAZOLE PESSARIES IP 2007
Chromatographic system Calculate the content of C22H17ClN2 in the cream.

– a stainless steel column 20 cm x 4.6 mm, packed with Storage. Store at a temperature not exceeding 30°.
30 volumes of 0.02 M phosphoric acid adjusted to pH
7.5 with a 10 per cent w/v solution of triethylamine in Clotrimazole Pessaries
methanol, Clotrimazole Vaginal Tablets
– spectrophotometer set at 215 nm, Clotrimazole Pessaries contain Clotrimazole in a suitable base.
– a 20 µl loop injector. Clotrimazole Pessaries contain not less than 95.0 per cent and
Inject reference solution (b).The column efficiency, determined not more than 105.0 per cent of the stated amount of
using the principal peak in the chromatogram obtained is not clotrimazole, C22H17ClN2.
less than 9000 theoretical plates.
Identification
Inject the test solution and reference solution (a). Allow the
chromatography to proceed for 1.5 times the retention time of A. Mix a quantity of the powdered pessaries containing 40 mg
the principal peak in the chromatogram obtained with the test of Clotrimazole with 20 ml of a mixture of 1 volume of 1 M
solution. The area of any peak corresponding to sulphuric acid and 4 volumes of methanol and shake with
2-chlorotritanol in the chromatogram obtained with the test two quantities, each of 50 ml, of carbon tetrachloride,
solution is not greater then the area of the peak in the discarding the organic layers. Make the aqueous phase alkaline
chromatogram obtained with reference solution (a). with dilute ammonia solution, add a further 5 ml of dilute
ammonia solution and extract with two quantities, each of
Other tests. Complies with the tests stated under Creams. 40 ml, of chloroform. Combine the chloroform extracts, shake
Assay. Determine by liquid chromatography (2.4.14). with 5 g of anhydrous sodium sulphate, filter and add sufficient
chloroform to the filtrate to produce 100 ml. Evaporate 50 ml
Test solution. Treat a quantity of the cream containing about
to dryness and dissolve the residue in 50 ml of a mixture of
20 mg of Clotrimazole as described in the test for
1 volume of 0.1 M hydrochloric acid and 9 volumes of
2-Chlorotritanol and dilute 1.0 ml of the filtrate to 5.0 ml with a
methanol. When examined in the range 230 nm to 360 nm, the
mixture of 70 volumes of methanol and 30 volumes of 0.02 M
resulting solution shows absorption maxima at about 262 nm
phosphoric acid.
and 265 nm (2.4.7).
Reference solution. Dissolve 20 mg of clotrimazole RS in 70 B. Determine by thin-layer chromatography (2.4.17), coating
ml of methanol, add sufficient 0.02 M phosphoric acid to the plate with silica gel G.
produce 100.0 ml and dilute 1.0 ml of the resulting solution to
5.0 ml with a mixture of 70 volumes of methanol and 30 volumes Mobile phase. Di-isopropyl ether in a chromatography tank,
of 0.02 M phosphoric acid. containing 25 ml of strong ammonia solution in a beaker.
Chromatographic system Test solution. Shake a quantity of the powdered pessaries

– a stainless steel column 25 cm x 4.6 mm, packed with containing 20 mg of Clotrimazole with 4 ml of dichloromethane
octadecylsilyl silica gel (5 µm), for 30 minutes, centrifuge and use the supernatant liquid.
– mobile phase: a mixture of 70 volumes of methanol and Reference solution. A 0.5 per cent w/v solution of clotrimazole
30 volumes of 0.02 M phosphoric acid adjusted to pH RS in dichloromethane.
7.5 with a 10 per cent w/v solution of triethylamine in Apply to the plate 10 µl of each solution. After development,
methanol, dry the plate in a current of air and spray with dilute potassium
– flow rate. 1.5 ml per minute, iodobismuthate solution. The spot in the chromatogram
– spectrophotometer set at 215 nm, obtained with the test solution is reddish brown and
– a 20 µl loop injector. corresponds to the spot in the chromatogram obtained with
Inject the reference solution. The column efficiency, determined the reference solution.
using the peak in the chromatogram obtained, should be not
less than 9000 theoretical plates. Tests
Inject alternately the test solution and the reference solution. Related substances. Determine by liquid chromatography
Allow the chromatography to proceed for 1.5 times the (2.4.14).
retention time of the principal peak in the chromatogram Test solution. Add 50 ml of methanol to a quantity of the
obtained with the test solution. powdered pessaries containing 0.1 g of Clotrimazole and shake
340
IP 2007 CLOXACILLIN SODIUM
for 20 minutes. Dilute to 100 ml with methanol and filter. To – a 20 µl loop injector.
20 ml of the filtrate add 50 ml methanol and sufficient 0.02 M Inject the reference solution. The column efficiency, determined
phosphoric acid to produce 100 ml. using the peak in the chromatogram obtained should be not
Reference solution (a). A 0.0002 per cent w/v solution of less than 9000 theoretical plates.
2-chlorotritanol RS in a mixture of 70 volumes of methanol
and 30 volumes of 0.02 M phosphoric acid.
Allow the chromatography to proceed for 1.5 times the
Reference solution (b). Dilute 1 volume of the test solution to retention time of the principal peak in the chromatogram
50 volumes with the same solvent mixture. obtained with the test solution.
Chromatographic system Calculate the content of C22H17ClN2 in the pessaries.
octadecylsilyl silica gel (5 µm), Storage. Store protected from moisture and crushing.
30 volumes of 0.02 M phosphoric acid adjusted to
pH 7.5 with a 10 per cent w/v solution of triethylamine
in methanol,
– flow rate. 1.5 ml per minute, Cloxacillin Sodium
COONa
Inject reference solution (b).The column efficiency, determined O
using the principal peak in the chromatogram obtained is not O N CH3
less than 9000 theoretical plates. ,H2O
S CH3
Cl N
Inject the test solution and reference solution (a). Allow the
N H H H
chromatography to proceed for 1.5 times the retention time of O CH3
the principal peak in the chromatogram obtained with the test
solution. The area of any peak corresponding to
2-chlorotritanol in the chromatogram obtained with the test C19H17CIN3NaO5S, H2O Mol. Wt. 475.9
solution is not greater then the area of the peak in the Cloxacillin Sodium is sodium (6R)-6-[3-(2-chlorophenyl)- 5-
chromatogram obtained with reference solution (a). methylisoxazole-4-carboxamido]penicillanate monohydrate.
Other tests. Complies with the tests stated under Pessaries. Cloxacillin Sodium contains not less than 95.0 per cent and
Assay. Determine by liquid chromatography (2.4.14). not more than 101.0 per cent of C19H17ClN3NaO5S, calculated
Test solution. Weigh and powder 20 pessaries. Weigh
accurately a quantity of the powder containing about 0.1 g of Description. A white or almost white, crystalline powder;
Clotrimazole, add 50 ml of methanol and shake for 20 minutes. hygroscopic.
Dilute to 250.0 ml with methanol and filter. To 10.0 ml of the
filtrate add 60 ml of methanol and sufficient 0.02 M phosphoric Identification
acid to produce 100.0 ml.
Reference solution. Dissolve 20 mg of clotrimazole RS in Compare the spectrum with that obtained with cloxacillin
70 ml of methanol, add sufficient 0.02 M phosphoric acid to sodium RS or with the reference spectrum of cloxacillin sodium.
produce 100.0 ml and dilute 1.0 ml of the resulting solution to
5.0 ml with a mixture of 70 volumes of methanol and 30 volumes
of 0.02 M phosphoric acid.
– a stainless steel column 25 cm x 4.6 mm, packed with C. Gives reaction A of sodium salts (2.3.1).
– mobile phase: a mixture of 70 volumes of methanol and Tests
30 volumes of 0.02 M phosphoric acid adjusted to Appearance of solution. A 10.0 per cent w/v solution in carbon
pH 7.5 with a 10 per cent w/v solution of triethylamine dioxide-free water is clear (2.4.1); absorbance of the solution
in methanol, at about 430 nm, not more than 0.04 (2.4.7).
– spectrophotometer set at 215 nm, pH (2.4.24). 5.0 to 7.0, determined in a 10.0 per cent w/v solution.
341
CLOXACILLIN CAPSULES IP 2007
Specific optical rotation (2.4.22). +160° to +169°, determined Cloxacillin Capsules

Cloxacillin Sodium Capsules
determined by Method A. Cloxacillin Capsules contain not less than 92.5 per cent and
not more than 107.5 per cent of the stated amount of cloxacillin,
C19H18ClN3O5S.
Identification
Buffer solution. Prepare a 0.02 M monobasic potassium
phosphate solution and adjust the pH to 6.6 with 2 M sodium A. Determine on the contents of the capsules by infrared
hydroxide. absorption spectrophotometry (2.4.6). Compare the spectrum
Test solution. Weigh accurately about 55 mg of the substance with that obtained with cloxacillin sodium RS or with the
under examination and dilute to 100.0 ml with the buffer reference spectrum of cloxacillin sodium.
solution. B. The contents of the capsules give reactions A and B of
Reference solution. Weigh accurately a suitable quantity of sodium salts (2.3.1).
cloxacillin sodium RS dissolve in the buffer solution and Tests
dilute to obtain a solution containing a known concentration
of about 0.55 mg per ml. Dissolution (2.5.2).
Chromatographic system Apparatus. No 1
– a stainless steel column 25 cm x 4.6 mm, packed with Medium. 900 ml of water
octadecylsilyl silica gel 3 to 10 µm, Speed and time. 100 rpm and 45 minutes.
– mobile phase: a mixture of 80 volumes of the buffer
solution and 20 volumes of acetonitrile,
not greater than 1.0 µm, rejecting the first 1 ml of the filtrate.
Dilute the filtrate, if necessary, with the same solvent. Measure
Inject the reference solution. The test is not valid unless the about 271 nm (2.4.7). Similarly measure the absorbance of a
tailing factor is not more than 1.8 and the relative standard standard solution of known concentration of cloxacillin
deviation for replicate injections is not more than 2.0 per cent. sodium RS in place of the contents of the capsules. Calculate
Inject alternately the test solution and the reference solution. the content of C19H18ClN3O5S.
Calculate the content of C19H18ClN3O5S. D. Not less than 75 per cent of the stated amount of
C19H18ClN3O5S.
Cloxacillin Sodium intended for use in the manufacture of
parenteral preparations without a further appropriate Other tests. Comply with the tests stated under Capsules.
procedure for the removal of bacterial endotoxins complies Assay. Determine by liquid chromatography (2.4.14).
Buffer solution. Prepare a 0.02 M monobasic potassium
Bacterial endotoxins (2.2.3). Not more than 0.40 Endotoxin phosphate solution and adjust the pH to 6.6 with 2 M sodium
Unit per mg of cloxacillin. hydroxide.
Cloxacillin Sodium intended for use in the manufacture of Test solution. Weigh accurately a quantity of the mixed
parenteral preparations without a further appropriate contents of 20 capsules containing about 50 mg of Cloxacillin,
sterilisation procedure complies with the following dissolve in the buffer solution by stirring for 10 minutes, dilute
additional requirement. to 100.0 ml with the buffer solution and filter.
Sterility (2.2.11). Complies with the test for sterility. Reference solution. Weigh accurately a suitable quantity of
Storage. Store protected from moisture at a temperature not cloxacillin sodium RS dissolve in the buffer solution and
exceeding 30°. If it is intended for use in the manufacture of dilute to obtain a solution containing a known concentration
parenteral preparations, the containers should be sterile and of about 0.55 mg per ml.
Labelling. The label states whether or not the material is – a stainless steel column 25 cm x 4.6 mm, packed with
intended for use in the manufacture of parenteral preparations. octadecylsilyl silica gel (3 to 10 µm),
342
IP 2007 CLOXACILLIN SYRUP
– mobile phase: a mixture of 80 volumes of the buffer Bacterial endotoxins (2.2.3). Not more than 0.40 Endotoxin
solution and 20 volumes of acetonitrile, Unit per mg of cloxacillin.
– flow rate. 1 ml per minute, Water (2.3.43). Not more than 4.5 per cent, determined on
– spectrophotometer set at 225 nm, 0.3 g.
tailing factor is not more than 1.8 and the relative standard Buffer solution. Prepare a 0.02 M monobasic potassium
deviation for replicate injections is not more than 2.0 per cent. phosphate solution and adjust the pH to 6.6 with 2 M sodium
Inject alternately the test solution and the reference solution. hydroxide.
Calculate the content of C19H18ClN3O5S in the capsules. Test solution. Determine the weight of the content of
10 containers. Weigh accurately a suitable quantity of the
Storage. Store protected from moisture at a temperature not mixed contents of the 10 containers containing about 55 mg of
exceeding 30°. Cloxacillin, dissolve in the buffer solution by shaking and
Labelling. The label states the strength in terms of the dilute to 100.0 ml with the buffer solution.
equivalent amount of cloxacillin.
cloxacillin sodium RS dissolve in the buffer solution and
dilute to obtain a solution containing a known concentration
Cloxacillin Injection of about 0.55 mg per ml.
Cloxacillin Sodium Injection Chromatographic system
Cloxacillin Injection is a sterile material consisting of Cloxacillin – a stainless steel column 25 cm x 4.6 mm, packed with
Sodium with or without excipients. It is filled in a sealed octadecylsilyl silica gel (3 to 10 µm),
container. – mobile phase: a mixture of 80 volumes of the buffer
solution and 20 volumes of acetonitrile,
The injection is constituted by dissolving the contents of the – flow rate. 1 ml per minute,
sealed container in the requisite amount of sterile Water for – spectrophotometer set at 225 nm,
Injections, immediately before use. – a 20 µl loop injector.
The constituted solution complies with the requirements for Inject the reference solution. The test is not valid unless the
Clarity of solution and Particulate matter stated under tailing factor is not more than 1.8 and the relative standard
Parenteral Preparations (Injections). deviation for replicate injections is not more than 2.0 per cent.
recommended by the manufacturer. Calculate the content of C19H18ClN3O5S in the injection.
Cloxacillin Injection contains not less than 90.0 per cent and Storage. Store protected from moisture at a temperature not
not more than 110.0 per cent of the stated amount of cloxacillin, exceeding 30°.
C19H18ClN3O5S. Labelling. The label states the quantity of Cloxacillin Sodium
Description. A white or almost white powder; very contained in the sealed container in terms of the equivalent
hygroscopic. amount of cloxacillin.
Cloxacillin Syrup
Identification Cloxacillin Sodium Syrup; Cloxacillin Elixir; Cloxacillin Sodium
A. Determine by infrared absorption spectrophotometry (2.4.6). Elixir; Cloxacillin Oral Solution; Cloxacillin Sodium Oral Solution
Compare the spectrum with that obtained with cloxacillin Cloxacillin Syrup is a mixture consisting of Cloxacillin Sodium
sodium RS or with the reference spectrum of cloxacillin sodium. with buffering agents and other excipients. It contains a
B. Gives reaction A of sodium salts (2.3.1). suitable flavouring agent. It is filled in a sealed container.
Tests the sealed container in the specified volume of Water just
pH (2.4.24). 5.0 to 7.0, determined in a 10 per cent w/v solution. before use.
343
CODEINE PHOSPHATE IP 2007
Cloxacillin Syrup contains not less than 90.0 per cent and not Labelling. The label states the strength in terms of the
more than 120.0 per cent of the stated amount of C19H18ClN3O5S. equivalent amount of cloxacillin.
When stored at the temperature and for the period stated on
the label during which the constituted suspension may be
expected to be satisfactory for use, it contains not less than
80.0 per cent of the stated amount of cloxacillin, C19H18ClN3O5S. Codeine Phosphate
Identification Codeine Phosphate Hemihydrate

with the test solution corresponds to the peak in the H3CO
The constituted suspension complies with the tests stated O
under Oral liquids and with the following tests. N ,H3PO4, 1/2H2O
H CH3
Tests
HO
pH (2.4.24). 4.0 to 7.0.
C18H21NO3,H3PO4,½ H2O Mol. Wt. 406.4
Buffer solution. Prepare a 0.02 M monobasic potassium Codeine Phosphate is (5R,6S)-7,8-didehydro-4,5-epoxy-3-
phosphate solution and adjust to pH 6.6 with 1 M sodium methoxy-17-methylmorphinan-6-ol dihydrogen phosphate
hydroxide. hemihydrate, an alkaloid occurring in Papaver somniferum.
Test solution. Transfer an accurately weighed quantity of the Codeine Phosphate contains not less than 98.5 per cent and
oral suspension containing about 125 mg of the cloxacillin to not more than 101.0 per cent of C18H21NO3,H3PO4, calculated
a 250-ml volumetric flask and dissolve in the buffer solution on the dried basis.
by stirring for 15 minutes. Dilute to 250.0 ml with the buffer Description. Colourless crystals or a white, crystalline powder.
solution.
Reference solution. Weigh accurately a suitable quantity of Identification
cloxacillin sodium RS dissolve in the buffer solution and Test A may be omitted if tests B, C, D, E and F are carried out.
dilute to obtain a solution containing a known concentration Tests B, C, D and E may be omitted if tests A and F are carried
of about 0.55 mg per ml. out.
Chromatographic system A. Dissolve 0.2 g in 4 ml of water, add 2 ml of 2 M sodium
– a stainless steel column 25 cm x 4.6 mm, packed with hydroxide and induce crystallisation, if necessary by
octadecylsilyl silica gel (3 to 10 µm), scratching the wall of the tube with a glass rod and cooling in
– mobile phase: a mixture of 80 volumes of the buffer ice. The residue after washing with water and drying at 100°
solution and 20 volumes of acetonitrile, to 105° complies with the following test.
– spectrophotometer set at 225 nm, Determine by infrared absorption spectrophotometry (2.4.6).
– a 20 µl loop injector. Compare the spectrum with that obtained with codeine
phosphate RS treated in the same manner or with the reference
Inject the reference solution. The test is not valid unless the spectrum of codeine.
deviation for replicate injections is not more than 2.0 per cent. B. To 25 ml of a 0.04 per cent w/v solution add 25 ml of water
and 10 ml of 1 M sodium hydroxide and dilute to 100 ml of
water. When examined in the range 230 nm to 360 nm, the
Determine the weight per ml of the oral suspension (2.4.29) resulting solution shows an absorption maximum only at about
and calculate the content of C19H18ClN3O5S weight in volume. 284 nm; absorbance at about 284 nm, about 0.38 (2.4.7).
Repeat the procedure using a portion of the constituted C. On the surface of one drop of nitric acid place a little of the
suspension that has been stored at the temperature and for powder; a yellow but not red colour is produced (distinction
the period stated on the label. from morphine).
Storage. Store protected from moisture, at a temperature not D. Dissolve 0.1 g in 1 ml of sulphuric acid and 1 drop of ferric
exceeding 30°. chloride test solution and warm gently; a bluish violet colour
344
IP 2007 CODEINE SYRUP
is produced. Add 1 drop of dilute nitric acid; the colour Assay. Weigh accurately about 0.4 g of the substance under
changes to red. examination and dissolve in a mixture of 10 ml of anhydrous
E. Gives the reaction of alkaloids (2.3.1). glacial acetic acid and 20 ml of dioxan. Titrate with 0.1 M
perchloric acid, using a few drops of crystal violet solution
F. Gives reaction A of phosphates (2.3.1). as indicator. Carry out blank titration.
Tests 1 ml of 0.1 M perchloric acid is equivalent to 0.03974 g of

C18H21NO3,H3PO4.
Appearance of solution. A 4.0 per cent w/v solution in carbon Storage. Store protected from light.
dioxide-free water prepared from distilled water is clear (2.4.1),
and not more intensely coloured than reference solution YS6
(2.4.1).
Codeine Syrup
Codeine Phosphate Syrup
Specific optical rotation (2.4.22). –98.0° to –102°, determined
in a 2.0 per cent w/v solution. Codeine Syrup is a 0.5 per cent w/v solution of Codeine
Phosphate in a suitable flavoured vehicle.
Foreign alkaloids. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel G. Codeine Syrup contains not less than 0.48 per cent w/v and
not more than 0.52 per cent w/v of codeine phosphate,
Mobile phase. A mixture of 72 volumes of ethanol, 30 volumes C18H21NO3,H3PO4, ½ H2O.
of cyclohexane and 6 volumes of strong ammonia solution.
Test solution. Dissolve 0.5 g of the substance under Identification
examination in sufficient of a mixture of 4 volumes of 0.01 M To 10 ml of the syrup add sufficient dilute ammonia solution
hydrochloric acid and 1 volume of ethanol to produce 10 ml. until the solution is alkaline and extract with three quantities,
Reference solution (a). Dilute 1.5 ml of the test solution to each of 10 ml, of chloroform. Evaporate the combined
100 ml with the same solvent. chloroform extracts to dryness on a water-bath and dry the
residue at 80°. The residue complies with the following tests.
Reference solution (b). Dilute 1 ml of the test solution to
100 ml with the same solvent. A. Dissolve 0.2 g in 4 ml of water, add 2 ml of 2 M sodium
hydroxide and induce crystallisation, if necessary by
Apply separately to the plate 10 µl of each solution. After scratching the wall of the tube with a glass rod and cooling in
development, dry the plate in air and spray with acidic ice. The residue washed with water and dried at 100° to 105°
potassium iodobismuthate solution. Any secondary spot in complies with the following test.
Compare the spectrum with that obtained with codeine
reference solution (a) and not more than one such spot, with
phosphate RS treated in the same manner or with the reference
an Rf value higher than that of the principal spot, is more
spectrum of codeine.
reference solution (b). B. Dissolve 0.1 g in 1 ml of sulphuric acid and 1 drop of ferric
chloride test solution and warm gently; a bluish violet colour
Chlorides (2.3.12). 12.5 ml of a 2.0 per cent w/v solution in
is produced. Add 1 drop of dilute nitric acid; the colour
distilled water complies with the limit test for chlorides
changes to red.
(0.1 per cent).
Sulphates (2.3.17). 7.5 ml of a 2.0 per cent w/v solution in Tests
distilled water complies with the limit test for sulphates Related substances. Determine by thin-layer chromatography
(0.1 per cent). (2.4.17), coating the plate with silica gel G.
Morphine. Dissolve 0.1 g in sufficient 0.1 M hydrochloric Mobile phase. A mixture of 72 volumes of ethanol, 30 volumes
acid to produce 5 ml, add 2 ml of a 1 per cent w/v solution of of cyclohexane and 6 volumes of strong ammonia solution.
sodium nitrite, allow to stand for 15 minutes and add 3 ml of
Test solution. To 20 ml of the syrup add 20 ml of water and 2 ml
6 M ammonia. The resulting solution is not more intensely
of strong ammonia solution and extract with two quantities,
each of 20 ml, of chloroform. Dry the combined extracts with
Loss on drying (2.4.19). Not more than 3.0 per cent, determined anhydrous sodium sulphate, filter, evaporate the filtrate to
on 1.0 g by drying in an oven at 105°. dryness and dissolve the residue in 1 ml of chloroform.
345
COLCHICINE IP 2007
Reference solution (a). Dilute 1.5 ml of the test solution to Description. Pale yellow crystals, amorphous scales or a
100 ml with the same solvent. powder.
Reference solution (b). Dilute 1 ml of the test solution to Identification
100 ml with the same solvent.
Apply separately to the plate 10 µl of each solution. After
development, dry the plate in air and spray with acidic
potassium iodobismuthate solution. Any secondary spot in A. Determine by infrared absorption spectrophotometry (2.4.6).
the chromatogram obtained with the test solution is not more Compare the spectrum with that obtained with colchicine RS
intense than the spot in the chromatogram obtained with or with the reference spectrum of colchicine.
reference solution (a) and not more than one such spot, with B. When examined in the range 230 nm to 400 nm, a 0.001 per
an Rf value higher than that of the principal spot, is more cent w/v solution in ethanol (95 per cent) shows absorption
intense than the spot in the chromatogram obtained with maxima, at about 243 nm and 350 nm; absorbance at about
reference solution (b). 243 nm, about 0.73 and at about 350 nm, about 0.42 (2.4.7).
Other tests. Complies with the tests stated under Oral Liquids. C. Dissolve 50 mg in 1.5 ml of water; a yellow solution is
Assay. Weigh accurately about 10.0 g, add dilute ammonia produced the colour of which is intensified on adding mineral
solution until the solution is alkaline to litmus paper and acids.
extract with four quantities, each of 25 ml, of chloroform. Wash D. Mix 1 mg with 0.2 ml of sulphuric acid in a white dish; a
each extract successively with the same 10 ml of water; combine yellow colour is produced which on the addition of 0.05 ml of
the chloroform extracts and evaporate to dryness on a water- nitric acid changes to greenish-blue and then rapidly becomes
bath. To the residue add 5 ml of ethanol (95 per cent) and reddish and finally almost colourless. On addition of an excess
again evaporate to dryness. Dissolve the residue in 5.0 ml of of 5 M sodium hydroxide the colour changes to red.
0.05 M hydrochloric acid and titrate the excess of acid with
0.05 M sodium hydroxide using methyl red solution as Tests
indicator. Specific optical rotation (2.4.22). -235° to -250°, determined at
1 ml of 0.05 M hydrochloric acid is equivalent to 0.02032 g of 20° in a 0.5 per cent w/v solution in ethanol (95 per cent).
C18H21NO3,H3PO4,½ H2O. Colchiceine. To 5 ml of a 1.0 per cent w/v solution add 0.1 ml
Determine the weight per ml of the syrup (2.4.29) and calculate of a 10.5 per cent w/v solution of ferric chloride. Any colour
the content of C18H21NO3,H3PO4 ,½ H2O, weight in volume. produced is not more intense than that obtained by mixing
2.0 ml of FCS with 1.0 ml of CCS and 2.0 ml of CSS (2.4.1).
(2.4.17), coating the plate with silica gel HF254.
Colchicine of 1,2-dichloroethane and 1 volume of strong ammonia
solution.
H3CO
NHCOCH3
H3CO
examination in 10 ml of chloroform.
H3CO
O
OCH3
Reference solution (b). Dilute 1 volume of reference solution
(a) with an equal volume of chloroform.
C22H25NO6 Mol. Wt. 399.4
Colchicine is (S)-N-(5,6,7,9-tetrahydro-1,2,3,10-tetramethoxy-
9-oxobenzo[a]heptalen-7-yl)acetamide, an alkaloid which
occurs in the corm and seeds of various species of Colchicum.
Colchicine contains not less than 97.0 per cent and not more chromatogram obtained with reference solution (a) and not
than 103.0 per cent of C22H25NO6, calculated on the anhydrous more than one such spot is more intense than the spot in
and solvent-free basis. chromatogram obtained with reference solution (b).
346
IP 2007 COLCHICINE TABLETS
Solvent. Determine by gas chromatography (2.4.14). Determine by infrared absorption spectrophotometry (2.4.6).
Test solution. A 1.0 per cent w/v solution of the substance Compare the spedtrum with that obtained with colchicine RS
under examination and the same concentration of the internal or with the reference spectrum of colchicine.
standard as in the reference solution. B. To a quantity of the powdered tablets containing 1 mg of
Colchicine add 0.2 ml of sulphuric acid and mix; a yellow
Reference solution. A solution in water containing 0.1 per
colour is produced. On adding a drop of nitric acid the colour
cent v/v of ethanol-free chloroform, 0.1 per cent v/v of ethyl
changes to greenish-blue, then reddish and finally almost
acetate and either 0.1 per cent v/v (for the determination of
colourless. On the addition of an excess of 5 M sodium
ethyl acetate) or 0.02 per cent v/v (for the determination of
hydroxide the colour changes to red.
chloroform) of ethanol (internal standard).
Chromatographic system Tests
– a glass or stainless steel column 1.5 m x 4 mm, packed
with white diatomaceous earth (100 to 120 mesh) coated Related substances. Determine by thin-layer chromatography
with 10 per cent w/w of polyethylene glycol 1000, (2.4.17), coating the plate with silica gel HF254.
– temperature: Mobile phase. A mixture of 50 volumes of acetone, 25 volumes
column.75°, of 1,2-dichloroethane and 1 volume of strong ammonia
inlet port. 130°, solution.
detector. 150°,
– a flame ionisation detector at 150°, Prepare the following solutions immediately before use.
– flow rate. 30 ml per minute of the carrier gas. Test solution. Extract a quantity of the powdered tablets
Calculate the percentage w/w of ethyl acetate or chloroform, containing 5 mg of Colchicine with 5 ml of chloroform, filter
taking 0.901 g or 1.477 g, respectively, as the weight per ml at and evaporate the filtrate to dryness in a current of air. Dissolve
20° (2.4.29). the residue as completely as possible in 0.1 ml of ethanol
(95 per cent), centrifuge and use the supernatant liquid.
The sum of the contents of chloroform or ethyl acetate and
the percentage of Water is not more than 10 per cent. Reference solution (a). Dilute 1 volume of the test solution to
20 volumes with ethanol (95 per cent).
Reference solution (b). Dilute 1 volume of reference solution
Water (2.3.43). Not more than 2.0 per cent, determined on (a) with an equal volume of chloroform.
0.5 g.
Assay. Weigh accurately about 50 mg and dissolve in a mixture
of 10 ml of acetic anhydride and 20 ml of toluene. Titrate with
0.02 M perchloric acid, determining the end-point
1 ml of 0.02 M perchloric acid is equivalent to 0.007988 g of more than one such spot is more intense than the spot in
C22H25NO6. chromatogram obtained with reference solution (b).
Storage. Store protected from light. Uniformity of content. Comply with the test stated under
Tablets.
Protect the solutions from light throughout the test.
Colchicine Tablets Crush one tablet and transfer to a centrifuge tube with the aid
of 10 ml of ethanol. Shake for 30 minutes, centrifuge and decant
Colchicine Tablets contain not less than 90.0 per cent and not
the supernatant liquid. Wash the residue with small quantities
more than 110.0 per cent of the stated amount of colchicine,
of ethanol, combine the extract and washings and add
C22H25NO6.
sufficient ethanol to produce a solution containing 0.001 per
Identification cent w/v of Colchicine. Measure the absorbance of the
resulting solution at the maximum at about 350 nm (2.4.7).
A.To a quantity of the powdered tablets containing 10 mg of Calculate the content of C22H25NO6 in the tablet taking 425 as
Colchicine add 20 ml of water and mix well. Filter into a the specific absorbance at 350 nm.
separating funnel and extract with 30 ml of chloroform.
Evaporate the chloroform extract to dryness using moderate
heat. The residue complies with the following test. Assay. Protect the solutions from light throughout the test.
347
COLCHICINE AND PROBENECID TABLETS IP 2007
Weigh and powder 20 tablets. Weigh accurately a quantity of Weigh and powder 20 tablets. Weigh accurately a quantity of
the powder containing about 0.5 mg of Colchicine, add 10 ml the powder containing about 1 mg of Colchicine, shake with
of ethanol and shake for 30 minutes. Centrifuge and decant 75 ml of ethanolic sodium carbonate for 30 minutes, add
the supernatant liquid. Wash the residue with small quantities sufficient of ethanolic sodium carbonate to produce 100.0 ml
of ethanol and mix the extract and washings. Add sufficient and filter. Measure the absorbance of the resulting solution at
ethanol to produce 50.0 ml and mix well. Measure the the maximum at about 350 nm (2.4.7). Calculate the content of
absorbance of the resulting solution at the maximum at about C22H25NO6 taking 425 as the specific absorbance at 350 nm.
350 nm (2.4.7). Calculate the content of C22H25NO6 taking 425 For probenecid — Weigh and powder 20 tablets. Weigh
as the specific absorbance at 350 nm. accurately a quantity of the powder containing about 0.25 g
Storage. Store protected from light. of Probenecid, shake with 100 ml of 0.1 M sodium hydroxide
for 10 minutes, add sufficient of 0.1 M sodium hydroxide to
produce 250.0 ml, mix and filter. Dilute 10.0 ml of the filtrate to
Colchicine and Probenecid Tablets 100.0 ml with 0.1 M sodium hydroxide. Dilute further 10.0 ml
to 50.0 ml with the same solvent and measure the absorbance
Colchicine and Probenecid Tablets contain not less than of the resulting solution at the maximum at about 248 nm (2.4.7).
90.0 per cent and not more than 115.0 per cent of the stated Calculate the content of C13H19NO4S taking 332 as the specific
amount of colchicine, C22H25NO6, and not less than 90.0 per absorbance at 248 nm.
probenecid, C13H19NO4S. Storage. Store protected from light.
Identification
A.Determine by thin-layer chromatography (2.4.17), coating
the plate with silica gel G. Cortisone Acetate
Mobile phase. A mixture of 100 volumes of methanol and
1.5 volumes of strong ammonia solution. O
O
Test solution (a). Shake a quantity of the powdered tablets O CH3
H3C
containing 1 mg of Colchicine with 15 ml of water, mix, filter, O OH
extract the filtrate with 25 ml of chloroform and evaporate the H3C H
chloroform extract to a volume of about 1 ml.
Test solution (b). Shake a quantity of the powdered tablets H H
containing 10 mg of Probenecid with 10 ml of chloroform, O
allow to settle and decant the clear supernatant liquid.
C23H30O6 Mol. Wt 402.5
w/v of colchicine RS. Cortisone Acetate is 17α,21-dihydroxypregn-4-ene-3,11,20-
trione 21-acetate.
w/v of probenecid RS. Cortisone Acetate contains not less than 96.0 per cent and
not more than 104.0 per cent of C23H30O6, calculated on the
dried basis.
The principal spot in the chromatogram obtained with test Description. A white or almost white, crystalline powder;
solution (a) corresponds to that in the chromatogram obtained odourless.
with reference solution (a). Similarly, the principal spot in the
chromatogram obtained with test solution (b) corresponds to Identification
that in the chromatogram obtained with reference solution (b). Tests A and B may be omitted if tests C and D are carried out.
Uniformity of content. For colchicine - Comply with the test Tests C and D may be omitted if tests A and B are carried out.
stated under Tablets using the method described under Assay. A. Determine by infrared absorption spectrophotometry (2.4.6).
Other tests. Comply with the tests stated under Tablets. Compare the spectrum with that obtained with cortisone
Assay. For colchicine — Carry out the determination acetate RS or with the reference spectrum of cortisone acetate.
without delay, under subdued light, using low actinic B. Determine by thin-layer chromatography (2.4.17), coating
glassware. the plate with silica gel G.
348
IP 2007 CORTISONE INJECTION
Solvent mixture. A mixture of 90 volumes of acetone and Chromatographic system

10 volumes of formamide. – a stainless steel column 25 cm x 4.6 mm, packed with
packed with octadecylsilyl silica gel (5 µm),
Mobile phase. Chloroform.
– mobile phase: a mixture of 400 ml of acetonitrile and
Test solution. Dissolve 25 mg of the substance under 550 ml of water, allowed to equilibrate sufficient water
examination in 10 ml of the same solvent mixture. added to produce 1000 ml and mixed,
Reference solution (a). Dissolve 25 mg of cortisone acetate
RS in 10 ml of the same solvent mixture.
Reference solution (b). Mix equal volumes of the test solution Equilibrate the column with the mobile phase for 30 minutes.
Inject reference solution (b). Adjust the sensitivity of the
Place the dry plate in a tank containing a shallow layer of the system so that the height of the principal peak in the
solvent mixture, allow the solvent mixture to ascend to the chromatogram is at least 50 per cent of the full scale of the
top, remove the plate from the tank and allow the solvent to recorder.
evaporate. Use within 2 hours, with the flow of the mobile
phase in the direction in which the aforementioned treatment Inject reference solution (a). The retention times are:
was done. hydrocortisone acetate, about 10 minutes and cortisone
acetate, about 12 minutes. The test is not valid unless the
Apply to the plate 2 µl of each solution. Allow the mobile resolution between the peaks due to hydrocortisone acetate
phase to rise 12 cm. Dry the plate in a current of warm air, allow and cortisone acetate is at least 4.2.
the solvent to evaporate, heat at 120° for 15 minutes and spray
Inject the test solution and reference solution (b). Continue
the hot plate with ethanolic sulphuric acid (20 per cent v/v).
the chromatography for twice the retention time of the principal
peak. In the chromatogram obtained with the test solution,
the area of any peak other than the principal peak, is not
greater than half the area of the principal peak in the
corresponds to that in the chromatogram obtained with
cent); the sum of the areas of all the peaks other than the
principal peak, is not greater than 1.5 times the area of the
compact spot.
D. 10 mg gives the reactions of acetyl groups (2.3.1). solution (b) (1.5 per cent). Ignore any peak with an area less
than 0.05 times the area of the principal peak in the
Tests chromatogram obtained with reference solution (b).
Specific optical rotation (2.4.22). +211° to +220°, determined Sulphated ash (2.3.18). Not more than 0.1 per cent.
in a 1.0 per cent w/v solution in dioxan. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Light absorption (2.4.7). Dissolve 50 mg in sufficient ethanol on 0.5 g by drying in an oven at 105° for 3 hours.
(95 per cent) to produce 100 ml and dilute 2 ml to 100 ml with Assay. Dissolve 0.1 g in ethanol and dilute to 100.0 ml with the
the same solvent. Absorbance of the resulting solution at the same solvent. Dilute 2.0 ml of the solution to 100.0 ml with
maximum at about 240 nm, 0.375 to 0.405. ethanol. Measure the absorbance at the maximum at about
237 nm (2.4.7).
(2.4.14). Calculate the content of C23H30O6 taking 395 as the specific
Test solution. Dissolve 25.0 mg of the substance under
examination in acetonitrile and dilute to 10.0 ml with the same
solvent.
Cortisone Injection
RS and 2 mg of hydrocortisone acetate RS in acetonitrile and Cortisone Acetate Injection
Cortisone Injection is a sterile suspension of a very fine powder
Reference solution (b). Dilute 1.0 ml of the test solution to of Cortisone Acetate in Sodium Chloride Injection containing
100.0 ml with acetonitrile. suitable dispersing agents.
349
CORTISONE INJECTION IP 2007
Cortisone Injection contains not less than 90.0 per cent and Test solution. Mix a quantity of the suspension containing
not more than 110.0 per cent of the stated amount of cortisone 25 mg of Cortisone Acetate with 15 ml of isopropyl alcohol,
acetate, C23H30O6. evaporate to dryness on a steam bath. To the residue add
Description. A white suspension, which settles on standing, 10 ml of the mobile phase, shake, mix with the aid of ultrasound
but readily disperses on shaking. and filter (such as Whatman GF/C filter).
Identification 100 volumes with the mobile phase.
Extract a volume of the injection containing 0.1 g of Cortisone Reference solution (b). A solution containing 0.002 per cent
Acetate with 6 ml of chloroform, filter and evaporate the w/v each of cortisone acetate RS and hydrocortisone acetate
chloroform. The residue complies with the following tests. RS in the mobile phase.
A. Determine by infrared absorption spectrophotometry (2.4.6). Chromatographic system

Compare the spectrum with that obtained with cortisone – a stainless steel column 25 cm x 4.6 mm, packed with
acetate RS or with the reference spectrum of cortisone acetate. packed with octadecylsilyl silica gel (5 µm) (such as
Hypersil ODS),
B. Determine by thin-layer chromatography (2.4.17), coating – mobile phase: a mixture of 400 ml of acetonitrile and
the plate with silica gel G. 550 ml of water, allowed to equilibrate and sufficient
Solvent mixture. A mixture of 90 volumes of acetone and water added to produce 1000 ml,
10 volumes of formamide. – flow rate. 1 ml per minute,
Mobile phase. Chloroform. – a 20 µl loop injector.
Test solution. Dissolve 25 mg of the substance under Inject reference solution (b). The retention times are:
examination in 10 ml of the solvent mixture. hydrocortisone acetate, about 10 minutes and cortisone
Reference solution (a). Dissolve 25 mg of cortisone acetate acetate, about 12 minutes. The test is not valid unless the
RS in 10 ml of the solvent mixture. resolution between the peaks due to hydrocortisone acetate
and cortisone acetate is at least 4.2.
Reference solution (b). Mix equal volumes of the test solution
and reference solution (a). Inject the test solution and reference solution (a). Continue
Place the dry plate in a tank containing a shallow layer of the peak. In the chromatogram obtained with the test solution,
solvent mixture, allow the solvent mixture to ascend to the the area of any secondary peak is not greater than half the
top, remove the plate from the tank and allow the solvent to area of the principal peak in the chromatogram obtained with
evaporate. Use within 2 hours, with the flow of the mobile reference solution (a) (0.5 per cent); the sum of the areas of all
phase in the direction in which the aforementioned treatment the secondary peaks is not greater than 1.5 times the area of
was done. the principal peak in the chromatogram obtained with reference
Apply to the plate 2 µl of each solution. Allow the mobile solution (a) (1.5 per cent). Ignore any peak with an area less
phase to rise 12 cm. Dry the plate in a current of warm air, allow than 0.05 times the area of the principal peak in the
the solvent to evaporate, heat at 120° for 15 minutes and spray chromatogram obtained with reference solution (a) (0.05 per
the hot plate with ethanolic sulphuric acid (20 per cent v/v). cent).
Heat at 120° for a further 10 minutes, allow to cool and examine Other tests. Complies with the tests stated under Parenteral
in daylight and in ultraviolet light at 365 nm. The principal Preparations (Injections).
reference solution (a). The principal spot in the chromatogram Test solution. Add 50 ml of methanol to a quantity of the
obtained with reference solution (b) appears as a single, injection containing about 10 mg of Cortisone Acetate, shake,
compact spot. mix with the aid of ultrasound for 2 minutes, dilute to 100.0 ml
with water, shake, centrifuge and use the supernatant liquid.
C. 10 mg gives the reactions of acetyl groups (2.3.1).
Reference solution. Dilute 50 ml of a solution in methanol
Tests containing 0.02 per cent w/v each of cortisone acetate RS and
prednisolone RS to 100.0 ml with water.
pH (2.4.24). 5.0 to 7.2.
Related substances. Determine by liquid chromatography – a stainless steel column 25 cm x 4.6 mm, packed with
(2.4.14). octadecylsilyl silica gel (5 µm) (such as Hypersil ODS),
350
IP 2007 CORTISONE TABLETS
– mobile phase: methanol (60 per cent), evaporate. Use within 2 hours, with the flow of the mobile
– flow rate. 1.5 ml per minute, phase in the direction in which the aforementioned treatment
– spectrophotometer set at 240 nm, was done.
– a 20 µl loop injector. Apply to the plate 2 µl of each solution. Allow the mobile
Inject the reference solution. The test is not valid unless the phase to rise 12 cm. Dry the plate in a current of warm air, allow
resolution between the peaks due to cortisone acetate and the solvent to evaporate, heat at 120° for 15 minutes and spray
prednisolone in the chromatogram obtained is at least 5.0. the hot plate with ethanolic sulphuric acid (20 per cent v/v).
Calculate the content of C23H30O6 in the injection. spot in the chromatogram obtained with the test solution
Storage. Store protected from light in single dose or multiple corresponds to that in the chromatogram obtained with
dose containers at a temperature not exceeding 30°. It should reference solution (a). The principal spot in the chromatogram
not be allowed to freeze. obtained with reference solution (b) appears as a single,
compact spot.
Labelling. The label states (1) the name(s) of the dispersing
agent(s) added; (2) that it is not meant to be given by C. 10 mg gives the reactions of acetyl groups (2.3.1).
intravenous injection; (3) that the container should be gently
shaken before a dose is withdrawn. Tests
(2.4.14).
Cortisone Tablets Test solution. Mix a quantity of the powdered tablets
containing 25 mg of Cortisone Acetate with 10 ml of the mobile
Cortisone Acetate Tablets phase, place in an ultrasonic bath for 10 minutes and filter
Cortisone Tablets contain not less than 90.0 per cent and not (such as Whatman GF/C filter).
more than 110.0 per cent of the stated amount of cortisone Reference solution (a). Dilute 1 volume of the test solution to
acetate, C23H30O6. 100 volumes with the mobile phase.
Identification Reference solution (b). A solution containing 0.002 per cent

w/v each of cortisone acetate RS and hydrocortisone acetate
Extract a quantity of the powdered tablets containing 0.1 g of RS in the mobile phase.
Cortisone Acetate with 5 ml of chloroform, filter and evaporate
the chloroform. The residue complies with the following tests.
A. Determine by infrared absorption spectrophotometry (2.4.6). packed with octadecylsilyl silica gel (5 µm) (such as
Compare the spectrum with that obtained with cortisone Hypersil ODS),
acetate RS or with the reference spectrum of cortisone acetate. – mobile phase: a mixture of 400 ml of acetonitrile and
B. Determine by thin-layer chromatography (2.4.17), coating 550 ml of water, allowed to equilibrate and sufficient
the plate with silica gel G. water added to produce 1000 ml,
Solvent mixture. A mixture of 90 volumes of acetone and – spectrophotometer set at 254 nm,
10 volumes of dimethylformamide. – a 20 µl loop injector.
Mobile phase. Chloroform. Inject reference solution (b). The retention times are:
Test solution. Dissolve 25 mg of the substance under hydrocortisone acetate, about 10 minutes and cortisone
examination in 10 ml of the same solvent mixture. acetate, about 12 minutes. The test is not valid unless the
resolution between the peaks due to hydrocortisone acetate
and cortisone acetate is at least 4.2.
RS in 10 ml of the same solvent mixture.
Inject the test solution and reference solution (a). Continue
Reference solution (b). Mix equal volumes of the test solution the chromatography for twice the retention time of the principal
and reference solution (a). peak. In the chromatogram obtained with the test solution,
Place the dry plate in a tank containing a shallow layer of the the area of any secondary peak is not greater than half the
solvent mixture, allow the solvent mixture to ascend to the area of the principal peak in the chromatogram obtained with
top, remove the plate from the tank and allow the solvent to reference solution (a) (0.5 per cent); the sum of the areas of all
351
ABSORBENT COTTON IP 2007
the secondary peaks is not greater than 1.5 times the area of Inject alternately the test solution and the reference solution.
the principal peak in the chromatogram obtained with reference Calculate the content of C23H30O6 in the tablets.
solution (a) (1.5 per cent). Ignore any peak with an area less
than 0.05 times the area of the principal peak in the Storage. Store protected from light.
chromatogram obtained with reference solution (a) (0.05 per
cent).
Uniformity of content. For tablets containing 10 mg or less. Absorbent Cotton
Comply with the test stated under Tablets. Absorbent Cotton Wool
Powder one tablet, add 50 ml of ethanol, shake for 30 minutes Absorbent Cotton consists of the new fibres or good quality
and add sufficient ethanol to produce 100.0 ml. Centrifuge new combers obtained from the seed coat of various species
and dilute a suitable volume of the supernatant liquid of the genus Gossypium Linn., cleaned, purified, bleached
containing 0.5 mg of Cortisone Acetate to 50.0 ml with ethanol. and carded. It does not contain any compensatory colouring
Measure the absorbance of the resulting solution at the matter.
maximum at about 240 nm (2.4.7). Calculate the content of Description. White, well-carded fibres of average staple length
C23H30O6 taking 390 as the specific absorbance at 240 nm. not less than 10 mm, containing not more than traces of leaf
Dissolution (2.5.2). residue, seed coat and other impurities. It offers appreciable
Apparatus. No 1 resistance when pulled and does not shed a significant
quantity of dust when shaken gently; practically odourless.
Medium. 900 ml of a mixture of a 1 per cent v/v solution of
hydrochloric acid and 30 volumes of 2-propanol Identification
A. When examined under a microscope, each fibre is seen to
Withdraw a suitable volume of the medium and filter through consist of a single cell, up to about 4 cm long and up to 40 µm
a membrane filter with an average pore diameter not greater wide, in the form of a flattened tube with thick and rounded
than 1.0 µm. Measure the absorbance of the filtrate, suitably walls and often twisted.
diluted if necessary with the medium, at the maximum at about
242 nm (2.4.7). Calculate the content of C23H30O6 taking 399 as B. Treat with iodinated zinc chloride solution; the fibres
the specific absorbance at 242 nm. become violet.
D. Not less than 75 per cent of the stated amount of C23H30O6. C. To 0.1 g add 10 ml of zinc chloride solution, heat to 40° and
allowed to stand for 2 ½ hours, shaking occasionally; the
Other tests. Comply with the tests stated under Tablets. fibres do not dissolve.
Tests
Test solution. Weigh and powder 20 tablets. Add 50 ml of
To 15.0 g add 150 ml of water, macerate for 2 hours in a closed
methanol to a quantity of the powder containing about 10 mg
vessel, decant the liquid, carefully squeezing out the residual
of Cortisone Acetate, shake, mix with the aid of ultrasound for
liquid with a glass rod and mix. Reserve 10 ml for the test for
2 minutes, dilute to 100.0ml with water, shake, centrifuge and
surface-active substances and filter the remainder (solution S).
use the supernatant liquid.
Acidity or alkalinity.To 25 ml of solution S add 0.1 ml of dilute
Reference solution. Dilute 50 ml of a solution in methanol
phenolphthalein solution; to another 25 ml add 0.05 ml of
containing 0.02 per cent w/v each of cortisone acetate RS and
methyl orange solution. Neither solution shows a pink colour.
prednisolone to 100.0 ml with water.
Surface-active substances. Into a 25-ml graduated, ground-
glass stoppered cylinder with external diameter of 18 to
22 mm, previously rinsed with sulphuric acid and then with
octadecylsilyl silica gel (5 µm) (such as Hypersil ODS),
water, add the 10-ml portion of solution S, shake vigorously
– mobile phase: methanol (60 per cent),
30 times in 10 seconds, allow to stand for 1 minute and shake
again 30 times. After 5 minutes, the height of the froth does
not exceed 2 mm above the surface of the liquid.
Inject the reference solution. The test is not valid unless the Absorbency
resolution between the peaks due to cortisone acetate and Apparatus. A dry, cylindrical wire basket, 80 mm high and
prednisolone in the chromatogram obtained is at least 5.0. 50 mm in diameter, fabricated from wire of diameter 0.4 mm and
352
IP 2007 CRESOL
having a mesh aperture of 15 to 20 mm; the basket weighs Evaporate 400 ml of the filtrate and dry the residue to constant
2.4 to 3.0 g. weight at 105°.
Sinking time. Not more than 10 seconds, determined by the Neps. A thin layer approximately equivalent to 0.5 g for an area
following method. of 450 sq. cm. spread uniformly between two glass plates, and
viewed by the naked eye under transmitted light, does not
Weigh the basket to the nearest 10 mg. Take five samples,
show more neps than about an average of 250 for three tests.
each of approximately 1 g, from different places in the material
being examined, pack loosely in the basket and weigh the Sulphated ash (2.3.18). Not more than 0.5 per cent, determined
packed basket to the nearest 10 mg. Hold the basket with its on 5.0 g.
long axis in the horizontal position and drop it from a height of Loss on drying (2.4.19). Not more than 8.0 per cent, determined
about 10 mm into water at 25° contained in a beaker at least on 5.0 g by drying in an oven at 105°.
12 cm in diameter and filled to a depth of 10 cm. Measure with
a stopwatch the time taken by the basket to sink below the Packaging. Package in rolls of not more than 500 g of
surface of the water. Repeat the procedure on two further continuous lap, with a light-weight paper running under the
samples and calculate the average value. entire lap, the paper being of such width that it may be folded
over the edges of the lap, the two together being tightly and,
Water-holding capacity. Not less than 23.0 g per g, determined evenly rolled, enclosed and sealed in a well-closed container.
by the following method.
Storage. Store protected from dust and moisture.
After the sinking time has been recorded in test A, remove the
basket from the water, allow it to drain for exactly 30 seconds
with its long axis in the horizontal position, transfer it to a
tared beaker and weigh to the nearest 10 mg. Calculate the Cresol
weight of water retained by the sample. Repeat the procedure
on two further samples and calculate the average value. Cresol is a mixture of cresols and other phenols obtained from
coal tar.
Foreign fibres. When examined under a microscope, it is seen
to consist exclusively of typical cotton fibres, except that Description. An almost colourless to pale brownish-yellow
occasionally a few isolated foreign fibres may be seen. liquid, becoming darker on keeping or on exposure to light;
odour, resembling that of phenol but more tarry.
Fluorescence. Examine a layer about 5 mm in thickness in
ultraviolet light at 365 nm. It shows only a slight, brownish- Identification
violet fluorescence and a few yellow particles. Not more than
a few isolated fibres show an intense blue fluorescence. To 0.5 ml add 300 ml of water, shake and filter. Divide the
filtrate into two parts. To one part add ferric chloride test
Colouring matter. Slowly extract 10 g in a narrow percolator solution; a transient bluish colour is produced. To the other
with ethanol (95 per cent) until 50 ml of extract is obtained. part add bromine solution; a pale yellow flocculent precipitate
The extract is not more intensely coloured than reference is produced.
solution YS5 or GYS6 (2.4.1) or a solution prepared in the
following manner. To 3.0 ml of CSS add 7.0 ml of a solution of Tests
hydrochloric acid containing 1 per cent v/v of hydrochloric
acid and dilute 0.5 ml of the resulting solution to 10 ml with Distillation range (2.4.8). Not more than 2.0 per cent v/v distils
the same solution of hydrochloric acid. below 188° and not less than 80.0 per cent v/v between 195°
and 205°.
Ether-soluble substances. Not more than 0.5 per cent
determined by the following method. Extract 5 g with ether in Acidity. A 2 per cent w/v solution is neutral to bromocresol
a continuous extraction apparatus (2.1.8), for 4 hours in such purple solution.
a way that the rate is at least four extractions per hour. Weight per ml (2.4.29). 1.029 g to 1.044 g.
Evaporate the ether and dry the residue to constant weight at
Hydrocarbons and volatile bases. Place 50 ml in a 500-ml round-
105°.
bottomed flask, add about 83 ml of a 27 per cent w/v solution
Water-soluble substances. Not more than 0.5 per cent, of sodium hydroxide and 100 ml of water and mix thoroughly.
determined by the following method. Boil 5 g with 500 ml of Connect the flask to a splash-bulb and air condenser about
water for 30 minutes, stirring frequently and replacing the 60 cm long, with the end of the air-condenser fitting closely
water lost by evaporation. Decant the liquid into a beaker, into the neck of a 250-ml pear-shaped separator and passing
squeeze the residual liquid from the material carefully with a well into the separator, which has a cylindrical graduated
glass rod, mix the liquids and filter the extract whilst hot. portion above the stopcock. Fill the graduated portion of the
353
CRESOL WITH SOAP SOLUTION IP 2007
separator with water. Distil rapidly until 75 ml of distillate has Tests

been collected, cooling the separator in running water, if
necessary. Allow the separator to stand in a vertical position Appearance of solution. 5 ml mixed with 95 ml of water forms
until separation is complete and draw off the aqueous liquid a clear solution without producing any opalescence on
into a titration flask. standing for 3 hours.
Alkalinity. Dilute 5 ml of ethanol (95 per cent) neutralised to
Hydrocarbons — Allow the separator to stand for a short
phenol red solution and titrate with 1 M sulphuric acid, using
time, measure the volume of hydrocarbon oil in the graduated
phenol red solution as indicator; not more than 0.6 ml is
portion and warm if necessary in order to keep the oil in the
required.
liquid state; subtract the volume of volatile bases in the
hydrocarbon oil, as determined in the following test; not more Hydrocarbons and volatile bases. Distil 120 ml until all the
than 0.5 per cent v/v of hydrocarbon oil is present. water and 50 ml of cresol have been collected. Place the cresol
thus recovered in a 500-ml round-bottomed flask, add about
Volatile bases — To the aqueous portion of the distillate
83 ml of a 27 per cent w/v solution of sodium hydroxide and
obtained in the preceding test, add any aqueous liquid still
100 ml of water and mix thoroughly. Connect the flask to a
remaining in the separator and neutralise it if necessary with
splash-bulb and air condenser about 60 cm long, with the end
0.1 M hydrochloric acid, using phenolphthalein solution as
of the air-condenser fitting closely into the neck of a 250-ml
indicator. Titrate with 1 M hydrochloric acid using methyl
pear-shaped separator and passing well into the separator,
orange solution as indicator. Wash the oil from the separator
which has a cylindrical graduated portion above the stopcock.
into the titration flask with water and again titrate with 1 M
Fill the graduated portion of the separator with water. Distil
hydrochloric acid. From the volume of additional 1 M
rapidly until 75 ml of distillate has been collected, cooling the
hydrochloric acid calculate the proportion of volatile bases
separator in running water, if necessary. Allow the separator
in the hydrocarbon oil. From the total volume of 1 M
to stand in a vertical position until separation is complete and
hydrochloric acid used in both titrations calculate the
draw off the aqueous liquid into a titration flask.
proportion of volatile bases in cresol.
Hydrocarbons — Allow the separator to stand for a short
1 ml of 1 M hydrochloric acid is equivalent to 0.08 ml of time, measure the volume of hydrocarbon oil in the graduated
volatile bases; not more than 0.15 per cent v/v of volatile portion and warm if necessary in order to keep the oil in the
bases, calculated as pyridine, are present. liquid state; subtract the volume of volatile bases in the
Sulphur compounds. Place about 20 ml in a small conical flask. hydrocarbon oil, as determined in the following test; not more
Moisten a piece of filter paper with a 10 per cent w/v solution than 0.5 per cent v/v of hydrocarbon oil is present.
of lead acetate and fix it on the mouth of the flask; heat the Volatile bases —To the aqueous portion of the distillate
flask on a water-bath for 5 minutes; the filter paper shows not obtained in the preceding test, add any aqueous liquid still
more than a light yellow colour. remaining in the separator and neutralise it if necessary with
Non-volatile matter. Not more than 0.1 per cent w/v when 0.1 M hydrochloric acid, using phenolphthalein solution as
evaporated on a water-bath and dried to constant weight at indicator. Titrate with 1 M hydrochloric acid using methyl
105°. orange solution as indicator. Wash the oil from the separator
into the titration flask with water and again titrate with 1 M
hydrochloric acid. From the volume of additional 1 M
hydrochloric acid calculate the proportion of volatile bases
in the hydrocarbon oil. From the total volume of 1 M
Cresol With Soap Solution hydrochloric acid used in both titrations calculate the
proportion of volatile bases in cresol.
Lysol
1 ml of 1 M hydrochloric acid is equivalent to 0.08 ml of
Cresol with Soap Solution is prepared by the saponification volatile bases; not more than 0.15 per cent v/v of volatile
of a mixture of Cresol with vegetable oils such as cotton seed, bases, calculated as pyridine, are present.
linseed, soyabean or similar oils but excluding coconut and
Sulphur compounds. Complies with the test for Sulphur
palm kernel oils. Alternatively, the mixed fatty acids derived
compounds described under Cresol.
from these oils may be used.
Assay. To 50 ml, accurately measured, add 150 ml of kerosene,
Cresol with Soap Solution contains not less than 47.0 per cent
mix and add little powdered pumice stone and 3 g of sodium
v/v and not more than 53.0 per cent v/v of Cresol.
bicarbonate. Distil into a separator, the rate of distillation
Description. An amber-coloured to reddish-brown liquid; being not more than 2 drops per second until the kerosene
odour, that of cresol; soapy to touch. and cresol have completely distilled. This is indicated by the
354
IP 2007 CROSCARMELLOSE SODIUM
distillate being yellow in colour. Stop the distillation, add addition. Titrate with 0.1 M sodium hydroxide until the colour
50 ml of kerosene and collect a further 50 ml of the distillate. turns to violet.
Discard the lower aqueous layer in the separator, dry the Calculate the number of milliequivalents ( M) of base required
remainder with anhydrous calcium chloride and shake with for the neutralisation equivalent to 1 g of dried substance.
10 ml of sulphuric acid (50 per cent w/w). Set aside for
2 hours, reject the acid layer and to the kerosene layer add Calculate the degree of acid carboxymethyl substitution (A)
40 ml of sodium hydroxide solution and shake for 5 minutes. from the expression:
Transfer the alkaline layer to a 100-ml volumetric flask and
1150 M
extract the kerosene layer with 20 ml of sodium hydroxide
solution adding the alkaline layer to that in the 100-ml (7102 − 412 M − 80 C )
volumetric flask. Add sodium hydroxide solution from a burette C = sulphated ash as a percentage
to make the volume in the flask to 100 ml. The difference
between the burette reading and 40.5 is equal to the volume of Calculate the degree of sodium carboxymethyl substitution
cresol in 50 ml of the sample. (S) from the expression:
Storage. Store protected from light. (162 + 58 A) C
(7102 − 80 C )
The degree of substitution is the sum of A + S and it is between
Croscarmellose Sodium 0.60 and 0.85, calculated on the dried basis.
Croscarmellose sodium (cross-linked sodium Sodium chloride and sodium glycollate. The sum of the
carboxymethylcellulose) is the sodium salt of a cross-linked, percentage contents of sodium chloride and sodium glycollate
partly O-carboxymethylated cellulose. is not more than 0.5 per cent, calculated on the dried basis.
Description. A white or greyish-white powder. Sodium chloride. Place 5.0 g in a 250 ml conical flask, add
50 ml of water and 5 ml of strong hydrogen peroxide solution
Identification and heat on a water-bath for 20 minutes stirring occasionally
to ensure total hydration. Cool, add 100 ml of water and 10 ml
A. Shake 1 g with 100 ml of 0.00001 per cent w/v solution of
of nitric acid. Titrate with 0.05 M silver nitrate determining
methylene blue and allow to settle. The substance under
the end-point potentiometrically (2.4.25) using a silver indicator
examination absorbs the methylene blue and settles as a blue,
electrode and a double-junction reference electrode
fibrous mass.
containing a 10 per cent w/v solution of potassium nitrate in
B. Shake 1 g with 50 ml of water. Transfer 1 ml of the mixture to the outer jacket and a standard filling solution in the inner
a test-tube, add 1 ml of water and 0.05 ml of a freshly prepared jacket, and stirring constantly.
4.0 per cent w/v solution of α-naphthol in methanol. Incline
the test-tube and add carefully 2 ml of sulphuric acid down
NaCl.
the side so that it forms a lower layer. A reddish-violet colour
develops at the interface. Sodium glycollate. Place 0.5 g of the substance under
examination in a 100 ml beaker. Add 5 ml of glacial acetic acid
C. The solution prepared from the sulphated ash in the test for and 5 ml of water and stir to ensure total hydration (about
Heavy metals (see Tests) gives reaction (a) of sodium salts 15 minutes). Add 50 ml of acetone and 1 g of sodium chloride.
(2.3.1) Stir for several minutes to ensure complete precipitation of
Tests the carboxymethylcellulose. Filter through a fast filter paper
impregnated with acetone into a volumetric flask, rinse the
pH (2.4.24). 5.0 to 7.0, determined on 1.0 per cent w/v solution beaker and filter with 30 ml of acetone and dilute the filtrate to
in carbon dioxide-free water. 100.0 ml with the same solvent. Allow to stand for 24 hours
Degree of substitution. Take 1.0 g in 500 ml conical flask, add without shaking. Use the clear supernatant to prepare the test
300 ml of a 10 per cent w/v solution of sodium chloride, solution.
25.0 ml of 0.1 M sodium hydroxide, stopper the flask and Reference solution. Dissolve 0.1 g of glycollic acid in 100 ml
allow to stand for 5 minutes, shaking occasionally. Add of water. Use the solution within 30 days. Transfer 1.0 ml,
0.05 ml of m-cresol purple solution and about 15 ml of 0.1 M 2.0 ml, 3.0 ml and 4.0 ml of the solution to separate volumetric
hydrochloric acid from a burette. Insert the stopper and shake. flasks; dilute the contents of each flask to 5.0 ml with water,
If the solution is violet, add 0.1 M hydrochloric acid in 1 ml add 5 ml of glacial acetic acid, dilute to 100.0 ml with acetone
portions until the solution becomes yellow, shaking after each and mix.
355
CROSPOVIDONE IP 2007
Transfer 2.0 ml of the test solution and 2.0 ml of each of the Sulphated ash (2.3.18). 14.0 to 28.0 per cent, determined on
reference solutions to separate 25 ml volumetric flasks. Heat 2.0 g, using a mixture of equal volumes of sulphuric acid and
the uncovered flasks for 20 minutes on a water-bath to eliminate water, and calculated on the dried basis.
acetone. Allow to cool and add 5.0 ml of 2,7- Loss on drying (2.4.19). Not more than 10.0 per cent, determined
dihydroxynaphthalene solution to each flask. Mix, add a on 1.0 g by drying in an oven at 105° for 6 hours.
further 15.0 ml of 2,7-dihydroxynaphthalene solution and
mix again. Close the flasks with aluminium foil and heat on a
water-bath for 20 minutes. Cool and dilute to 25.0 ml with
sulphuric acid. Crospovidone
Measure the absorbance (2.4.7) of each solution at 540 nm. l-Ethenyl-2-pyrrolidinone homopolymer; 1-Vinyl-2-
Prepare a blank using 2.0 ml of a solution containing 5 per cent pyrrolidinone homopolymer
v/v each of glacial acetic acid and water in acetone. Prepare
a standard curve using the absorbances obtained with the Crospovidone is a water-insoluble synthetic crosslinked
reference solutions. From the standard curve and the homopolymer of N-vinyl-2-pyrrolidinone.
absorbance of the test solution, determine the mass, in Crospovidone contains not less than 11.0 per cent and not
milligrams, of glycollic acid in the substance under examination, more than 12.8 per cent of nitrogen (N), calculated on the
and calculate the content of sodium glycollate from the anhydrous basis.
expression:
Description. A white to creamy white hygroscopic powder
10 × 1.29 × a having a faint odour.
(100 − b) m Identification
1.29 = the factor converting glycollic acid to sodium A. Determine by infrared absorption spectrophotometry (2.4.6)
glycollate on specimen previously dried in vacuum at 105° for 1 hour.
b = loss on drying as a percentage Compare the spectrum with that obtained with crospovidone
RS.
m = mass of the substance under examination, in
grams B. Suspend 1 g in 10 ml of water, add 0.1 ml of 0.1 M iodine,
and shake for 30 seconds. Add 1 ml of starch solution, and
Water-soluble substances. Not more than 10.0 per cent. shake; no blue color develops.
Disperse 10.0 g in 800.0 ml of water and stir for 1 minute every
10 minutes during the first 30 minutes. Allow to stand for Tests
1 hour and centrifuge, if necessary. Decant 200.0 ml of the
supernatant liquid onto a fast filter paper in a vacuum filtration pH (2.4.24). 5.0 and 8.0, determined in a 1.0 per cent w/v
funnel, apply vacuum and collect 150.0 ml of the filtrate. aqueous suspension.
Evaporate to dryness and dry the residue at 100° to 105° for Water (2.3.43). Not more than 0.4 per cent, determined on
4 hours. 2.0 g.
Heavy metals (2.3.13). To the residue obtained in sulphated Water-soluble substances. Transfer 25.0 g to a 400 ml beaker,
ash add 1 ml of hydrochloric acid and evaporate on a water- add 200 ml of water, and stir on a magnetic stirrer, using a 5-cm
bath. Take up the residue in 20 ml of water. 12 ml of the solution stirring bar, for 1 hour. Transfer to a 250 ml volumetric flask
complies with the limit test for heavy metals, Method A with the aid of about 25 ml of water, add water to volume, and
(10 ppm). Prepare the reference solution using lead standard mix. Allow the bulk of the solids to settle. Pass about 100 ml of
solution (1 ppm Pb). the relatively clear supernatant through a membrane filter
Settling volume. 10.0 to 30.0 ml. Place 75 ml of water in a having a 0.45 mm porosity, protected against clogging by
100 ml graduated cylinder and add 1.5 g of the substance super imposing a membrane filter. Transfer 50.0 ml of the clear
under examination in 0.5 g portions, shaking vigorously after filtrate to a tared 100 ml beaker, evaporate to dryness, and dry
each addition. Dilute to 100.0 ml with water and shake again at 110° for 3 hours: the weight of the residue does not exceed
until the substance is homogeneously distributed. Allow to 75 mg (1.5 per cent).
stand for 4 hours. Heavy metals (2.3.13). 2.0 g complies with the limit test for
Microbial contamination (2.2.9). Total microbial count is not
more than 103 bacteria and 102 fungi per gram, determined by Vinylpyrrolidinone. Suspend 4.0 g in 20 ml of water, stir for
plate count. 10 g is free from Escherichia coli. 15 minutes, centrifuge the suspension, and filter the slightly
356
IP 2007 CYANOCOBALAMIN
turbid upper layer through a sintered glass 10 mm filter. Stir Labelling. The label states the type (type A or type B).
the lower layer with 50 ml of water, centrifuge, and filter the
upper layer through the same filter. Again stir the lower layer
with 50 ml of water, and filter similarly. Add 0.5 g of sodium
acetate to the combined filtrates and titrate with 0.1 M iodine Cyanocobalamin
until the color of iodine no longer fades, add 3.0 ml of 0.1 M Vitamin B12
iodine, allow to stand for 10 minutes, and titrate the excess of
iodine with 0.1 M sodium thiosulphate, adding 3 ml of starch
solution as the end-point is approached. Carry out a blank H2NOCH2CH2C CH3 CH3
CH2CONH2
determination, using the same total volume of the same 0.1 M H2NOCH2C
CH2CH2CONH2
iodine, accurately measured, as was used for titrating the H3 C
N N
substance under examination. Before titrating the blank, adjust H3C Co N CH3
with acetic acid to the same pH as that of the substance H N N
CH3 N
under examination; not more than 0.72 ml of 0.1 M iodine is CH3
H2 NOCH2C CH3
HO H
consumed, corresponding to not more than 0.1 per cent of H3C CH3 CH2CH2CONH2
vinylpyrrolidinone. O
O O
Nitrogen. Place 0.1 g of the substance under examination O P O
CH2OH
N
(m mg) in a combustion flask, add 5 g of a mixture of 1 g of H O
CH3
copper sulphate, 1 g of titanium dioxide and 33 g of
dipotassium sulphate, and 3 glass beads. Wash any adhering
particles from the neck into the flask with a small quantity of C63H88CoN14O14P Mol. Wt. 1355.4
water. Add 7 ml of sulphuric acid, allowing it to run down the Cyanocobalamin is Coα-[α-(5,6-dimethylbenzimidazolyl)]-
sides of the flask, and mix the contents by rotation. Close the Coβ-cyanocobamide.
mouth of the flask loosely, for example by means of a glass
bulb with a short stem, to avoid excessive loss of sulphuric Cyanocobalamin contains not less than 96.0 per cent and not
acid. Heat gradually at first, then increase the temperature more than 102.0 per cent of C63H88CoN14O14P, calculated on
until there is vigorous boiling with condensation of sulphuric the dried basis.
acid in the neck of the flask; precautions are to be taken to Description. A dark red, crystalline powder; very hygroscopic.
prevent the upper part of the flask from becoming overheated.
Continue the heating for 45 minutes. Cool, dissolve the solid Identification
material by cautiously adding to the mixture 20 ml of water,
A. When examined in the range 260 nm to 610 nm, a 0.0025 per
cool again and place in a steam-distillation apparatus. Add
cent w/v solution shows absorption maxima, at about 278 nm,
30 ml of strong sodium hydroxide solution through the funnel,
361 nm and 547 nm to 559 nm. The ratio of the absorbance at
rinse the funnel cautiously with 10 ml of water and distil
about 361 nm to that at about 547nm to 559 nm is 3.15 to
immediately by passing steam through the mixture. Collect
3.45 and the ratio of the absorbance at about 361 nm to that at
80-100 ml of distillate in a mixture of 30 ml of a 4.0 per cent
about 278 nm is 1.70 to 1.90 (2.4.7).
w/v solution of boric acid and 0.05 ml of bromocresol green-
methyl red solution and enough water to cover the tip of the B. Determine by thin-layer chromatography (2.4.17), protected
condenser. Towards the end of the distillation lower the form light and coating the plate with silica gel G.
receiver so that the tip of the condenser is above the surface Mobile phase. A mixture of 60 volumes of chloroform,
of the acid solution and rinse the end part of the condenser 40 volumes of methanol and 12 volumes of 6 M ammonia.
with a small quantity of water. Titrate the distillate with Use an unlined tank.
0.025 M sulphuric acid until the colour of the solution changes
from green through pale greyish-blue to pale greyish-red- Test solution. Dissolve 20 mg of the substance under
purple (n1 ml of 0.025 M sulphuric acid). examination in 10 ml of ethanol (50 per cent).
Repeat the test using about 100 mg of glucose in place of the Reference solution. A 0.2 per cent w/v solution of
substance under examination (n2 ml of 0.025 M sulphuric cyanocobalamin RS in ethanol (50 per cent).
acid). Apply to the plate 10 µl of each solution. Allow the mobile
0.7004 (n 1 − n 2 ) phase to rise 12 cm. Dry the plate in air and examine in day
Percentage content of nitrogen = light. The principal spot in the chromatogram obtained with
m
the test solution corresponds to that in the chromatogram
Storage. Store protected from moisture. obtained with the reference solution.
357
CYANOCOBALAMIN INJECTION IP 2007
C. Mix about 1 mg with 10 mg of potassium sulphate and Loss on drying (2.4.19). Not more than 12.0 per cent,
0.1 ml of 0.5 M sulphuric acid and heat carefully to redness. determined on 20.0 mg by drying in an oven at 105° at a pressure
Allow to cool, add to the residue 0.1 ml of water, 0.5 ml of of 1.5 to 2.5 kPa for 2 hours.
saturated solution of ammonium thiocyanate and 0.5 ml of Assay. Weigh accurately about 25 mg and dissolve in sufficient
benzyl alcohol and shake; a blue colour is formed and is water to produce 1000.0 ml. Measure the absorbance of the
taken into the benzyl alcohol layer. solution at the maximum at about 361 nm (2.4.7). Calculate the
content of C 63H88CoN 14O 14P taking 207 as the specific
Tests absorbance at 361 nm.
Related substances. Determine by liquid chromatography Storage. Store protected from light and moisture.
(2.4.17).
The following solutions should be used within 1 hour of
preparation. Cyanocobalamin Injection
Test solution. Dissolve 10 mg of the substance under Vitamin B12 Injection
Cyanocobalamin Injection is a sterile solution of
Reference solution (a). A 0.003 per cent w/v solution of the Cyanocobalamin in Water for Injections containing sufficient
substance under examination in the mobile phase. Acetic Acid or Hydrochloric Acid to adjust the pH to about 4.
It may contain suitable buffering agents.
substance under examination in the mobile phase. Cyanocobalamin Injection contains not less than 95.0 per cent
Reference solution (c). Dissolve 5 mg of the substance under anhydrous cyanocobalamin, C63H88CoN14O14P.
examination in 2 ml of water, warming if necessary, allow to
cool, add 1 ml of a 0.1 per cent w/v solution of chloramine T Identification
and 0.1 ml of 0.05 M hydrochloric acid, dilute to 5 ml with
water, shake and allow to stand for 5 minutes. Dilute 1 ml of Measure the absorbance at about 278 nm, 361 nm and 550 nm.
this solution to 10 ml with the mobile phase. Use immediately. The ratio of the absorbance at about 278 nm to that at about
550 nm is 0.57 and the ratio of the absorbance at about 550 nm
Chromatographic system to that at about 361 nm is 0.30 (2.4.7).
octylsilyl silica gel (5 µm), Tests
pH (2.4.24). 3.8 to 5.5.
w/v solution of disodium hydrogen phosphate and
53 volumes of methanol adjusted to pH 3.5 with Related substances. Determine by liquid chromatography
phosphoric acid (to be used within 2 days of (2.4.14).
preparation), The following solutions should be used within 1 hour of
– flow rate. 0.8 ml per minute, preparation.
Test solution (a). Dilute a suitable volume of the injection, if
necessary, with the mobile phase to produce a solution
Inject the test solution and reference solutions (a), (b) and (c). containing 0.0001 per cent w/v of Cyanocobalamin.
Allow the chromatography to proceed for three times the Test solution (b). Dilute a suitable volume of the injection, if
retention time of the peak due to cyanocobalamin. necessary, with the mobile phase to produce a solution
containing 0.05 per cent w/v of Cyanocobalamin.
In the chromatogram obtained with the test solution the sum
of the areas of any secondary peaks is not greater than the Reference solution (a). Dilute a suitable volume of the
area of the principal peak in the chromatogram obtained with injection, if necessary, with the mobile phase to produce a
reference solution (a). Ignore any peak the area of which is solution containing 0.003 per cent w/v of Cyanocobalamin.
less than that of the principal peak in the chromatogram Reference solution (b). Add 1 ml of a 0.1 per cent w/v solution
obtained with reference solution (b). The test is not valid of chloramine T and 0.1 ml of 0.05 M hydrochloric acid to a
unless the chromatogram obtained with reference solution (c) volume containing 5 mg of Cyanocobalamin, dilute to 10 ml
exhibits two principal peaks, the resolution between these with water, shake and allow to stand for 5 minutes. Dilute 2 ml
peaks is 2.5 or more and the chromatogram obtained with of this solution to 10 ml with the mobile phase and use
reference solution (a) exhibits one principal peak. immediately.
358
IP 2007 CYCLIZINE HYDROCHLORIDE
Chromatographic system Cyclizine Hydrochloride contains not less than 98.0 per cent
– a stainless steel column 25 cm x 4 mm, packed with and not more than 101.0 per cent of C18H22N2,HCl, calculated
octylsiyl silica gel (5 µm), on the dried basis.
– mobile phase: a mixture of 147 volumes of a 1.0 per cent Description. A white, crystalline powder; almost odourless.
w/v solution of disodium hydrogen phosphate and
53 volumes of methanol, adjusted to pH 3.5 with
Identification
phosphoric acid (to be used within 2 days of
preparation), A.Determine by infrared absorption spectrophotometry (2.4.6).
– flow rate. 0.8 ml per minute, Compare the spectrum with that obtained with cyclizine
– spectrophotometer set at 361 nm, hydrochloride RS or with the reference spectrum of cyclizine
– a 20 µl loop injector. hydrochloride.
Allow the chromatography to proceed for three times the B. When examined in the range 220 nm to 360 nm, a freshly
retention time of the peak due to cyanocobalamin. prepared 0.002 per cent w/v solution in 0.05 M sulphuric acid
In the chromatogram obtained with test solution (b) the sum shows absorption maximum only at about 225 nm; absorbance
of the areas of any secondary peaks is not greater than the at about 225 nm, about 0.78 (2.4.7).
area of the principal peak in the chromatogram obtained with C. Dissolve 0.5 g in 10 ml of ethanol (95 per cent), warming if
reference solution (a). Ignore any peak the area of which is necessary, cool in ice, add 1 ml of 5 M sodium hydroxide and
less than that of the principal peak in the chromatogram sufficient water to produce 20 ml. Stir well and filter; the
obtained with test solution (a). precipitate, after washing with water and drying at 60° at a
Other tests. Complies with the tests stated under Parenteral pressure not exceeding 0.7 kPa for 2 hours, melts at about 107°
Preparations (Injections). (2.4.21).
Assay. Carry out the following procedure protected from D. Gives the reactions of chlorides (2.3.1).
light.
Tests
Dilute the injection, if necessary, with water to produce a
solution containing not more than the equivalent of N-Methylpiperazine. Determine by thin-layer chromatography
0.0025 per cent w/v of anhydrous cyanocobalamin and measure (2.4.17), coating the plate with silica gel G.
the absorbance at the maximum at about 361 nm (2.4.7).
Calculate the content of C63H88CoN14O14P taking 207 as the
solution.
Storage. Store protected from light in single dose or multiple
dose containers.
Labelling. The label states the strength in terms of the Test solution. Dissolve 0.1 g of the substance under
equivalent amount of anhydrous cyanocobalamin in a suitable examination in 10 ml of methanol.
dose-volume. Reference solution. A 0.005 per cent w/v solution of
N-methylpiperazine RS in methanol.
Cyclizine Hydrochloride dry the plate in air and expose to iodine vapours for 10 minutes.
CH3 test solution is not more intense than the corresponding spot
N in the chromatogram obtained with the reference solution.
N Sulphated ash (2.3.18). Not more than 0.1 per cent.
,HCl
anhydrous glacial acetic acid and add 50 ml of mercuric
C18H22N2, HCl Mol. Wt. 302.9
acetate solution. Titrate with 0.1 M perchloric acid,
Cyclizine Hydrochloride is 1-(diphenylmethyl)-4- determining the end-point potentiometrically (2.4.25). Carry
methylpiperazine hydrochloride out a blank titration.
359
CYCLIZINE TABLETS IP 2007
1 ml of 0.1 M perchloric acid is equivalent to 0.01514 g of 500.0 ml and filter. Dilute 5.0 ml of the filtrate to 100.0 ml with
C18H22N2,HCl. 0.5 M sulphuric acid and measure the absorbance of the
Storage. Store protected from light. resulting solution at the maximum at about 225 nm (2.4.7).
Calculate the content of C18H22N2,HCl taking 390 as the specific
Cyclizine Tablets
Cyclizine Hydrochloride Tablets
Cyclizine Tablets contain not less than 92.5 per cent and not Cyclophosphamide
more than 107.5 per cent of the stated amount of cyclizine
hydrochloride, C18H22N2,HCl. O Cl
O
P N
,H2O
Identification NH Cl
of Cyclizine Hydrochloride with 10 ml of ethanol (95 per cent), C7H15CI2N2O2P, H2O Mol. Wt. 279.1
filter and evaporate the filtrate to dryness. The residue complies Cyclophosphamide is (RS)-2-bis(2-
with the following test. chloroethyl)aminoperhydro-1,3,2-oxazaphosphorinane 2-
oxide monohydrate.
Compare the spectrum with that obtained with cyclizine Cyclophosphamide contains not less than 98.0 per cent and
hydrochloride RS or with the reference spectrum of cyclizine not more than 102.0 per cent of C7H15Cl2N2O2P, calculated on
hydrochloride. the anhydrous basis.
B. Extract a quantity of the powdered tablets containing 0.5 g Description. A white or almost white, crystalline powder.
of Cyclizine Hydrochloride with 20 ml of water and filter. The
filtrate gives reaction A of chlorides (2.3.1). Identification
Tests Test A may be omitted if tests B, C and D are carried out. Tests
N-Methylpiperazine. Determine by thin-layer chromatography
Mobile phase. A mixture of 90 volumes of chloroform, cyclophosphamide RS or with the reference spectrum of
8 volumes of methanol and 2 volumes of strong ammonia cyclophosphamide.
solution. B. In the test for Related substances, the principal spot in the
Prepare the following solutions freshly. chromatogram obtained with test solution (b) corresponds to
that in the chromatogram obtained with reference solution (b).
containing 0.1 g of Cyclizine Hydrochloride with 10 ml of C. Dissolve 0.1 g in 10 ml of water and add 5 ml of silver
methanol and filter. nitrate solution; no precipitate is produced. Boil; a white
precipitate is produced which is insoluble in dilute nitric acid
but is soluble in dilute ammonia solution from which it can be
N-methylpiperazine RS in methanol.
reprecipitated by the addition of dilute nitric acid.
dry the plate in air and expose to iodine vapours for 10 minutes. D. Dissolve 0.1 g in 3 ml of nitric acid and 1 ml of sulphuric
Any secondary spot in the chromatogram obtained with the acid, heat till brown fumes are evolved and the solution
test solution is not more intense than the corresponding spot becomes colourless. Cool, add 10 ml of water, heat again up to
in the chromatogram obtained with the reference solution. 60° and add 10 ml of ammonium molybdate solution; a bright
yellow precipitate is slowly formed.
Tests
quantity of the powder containing about 0.125 g of Cyclizine Appearance of solution. A 2.0 per cent w/v solution in carbon
Hydrochloride and shake with 400 ml of 0.5 M sulphuric acid dioxide-free water is clear (2.4.1), and not more intensely
for 15 minutes. Add sufficient 0.5 M sulphuric acid to produce coloured than reference solution YS6 (2.4.1).
360
IP 2007 CYCLOPHOSPHAMIDE INJECTION
pH (2.4.24). 4.0 to 6.0, determined in a 2.0 per cent w/v solution. endotoxins complies with the following additional
Related substances. Determine by thin-layer chromatography requirement.
(2.4.17), coating the plate with silica gel G. Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin
Mobile phase. A mixture of 80 volumes of 2-butanone, Unit per mg of cyclophosphamide.
12 volumes of water, 4 volumes of acetone and 2 volumes of Cyclophosphamide Phosphate intended for use in the
anhydrous formic acid. manufacture of parenteral preparations without a further
Test solution (a). Dissolve 0.2 g of the substance under appropriate sterilization procedure complies with the
examination in 10 ml of ethanol (95 per cent). following additional requirement.
Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with Sterlity. Complies with the test for sterility (2.2.11).
ethanol (95 per cent). Storage. Store in a refrigerator (2° to 8°). Avoid long exposure
Reference solution (a). Dilute 5 ml of test solution (b) to 50 ml to temperatures above 30°.
with ethanol (95 per cent). Labelling. The label states whether or not the material is
Reference solution (b). A 0.2 per cent w/v solution of intended for use in the manufacture of parenteral preparations.
cyclophosphamide RS in ethanol (95 per cent).
dry the plate in a current of warm air and heat at 110° for Cyclophosphamide Injection
10 minutes. Place the plate while hot in a tank in which is
placed a dish containing equal volumes of a 5 per cent w/v Cyclophosphamide for Injection is a sterile material consisting
solution of potassium permanganate and hydrochloric acid, of 100 parts by weight of Cyclophosphamide and 45 parts by
close the tank and allow to stand for 2 minutes. Remove the weight of Sodium Chloride. It is filled in a sealed container.
plate and place it in a current of cold air until excess chlorine is The injection is constituted by dissolving the contents of the
removed and an area of coating below the line of application sealed container in the requisite amount of sterile Water for
gives not more than a faint blue colour with potassium iodide Injections, immediately before use.
and starch solution; do not expose long to cold air. Spray the
plate with potassium iodide and starch solution and allow to
stand for 5 minutes. Any secondary spot in the chromatogram
obtained with test solution (a) is not more intense than the
spot in the chromatogram obtained with reference solution Storage. The constituted solution should be used immediately
(a). Ignore any spot remaining on the line of application. after preparation but, in any case, within the period
Heavy metals (2.3.13). 1.0 g dissolved in 2 ml of dilute acetic
acid and diluted to 25 ml with water complies with the limit Cyclophosphamide Injection contains not less than 92.5 per
test for heavy metals, Method A (20 ppm). cent and not more than 107.5 per cent of the stated amount of
cyclophosphamide, C7H15Cl2N2O2P.
Chlorides (2.3.12). A freshly prepared solution of 0.75 g in
sufficient water to produce 25 ml complies with the limit test Description. A white or almost white powder.
for chlorides (330 ppm). The contents of the sealed container comply with the
Water (2.3.43). 5.8 to 7.0 per cent, determined on 0.3 g. requirements stated under Parenteral Preparations
Assay. Weigh accurately about 0.1 g and dissolve in 50 ml of (Powders for Injection) and with the following requirements.
a 0.1 per cent w/v solution of sodium hydroxide in ethylene Identification
glycol; boil under a reflux condenser for 30 minutes and allow
to cool. Rinse the condenser with 25 ml of water, add 75 ml of A. Shake a quantity containing 0.2 g of anhydrous
2-propanol, 15 ml of 2 M nitric acid, 10.0 ml of 0.1 M silver cyclophosphamide with 2 ml of chloroform and filter. The
nitrate and 2 ml of ferric ammonium sulphate solution and solution complies with the following test.
titrate with 0.1 M ammonium thiocyanate. Determine by infrared absorption spectrophotometry (2.4.6).
1 ml of 0.1 M silver nitrate is equivalent to 0.01305 g of Compare the spectrum with that obtained with
C7H15Cl2N2O2P. cyclophosphamide RS or with the reference spectrum of
Cyclophosphamide Phosphate intended for use in the cyclophosphamide.
manufacture of parenteral preparations without a further B. Extract a quantity containing 0.2 g of anhydrous
appropriate procedure for the removal of bacterial cyclophosphamide with ether and evaporate the extract to
361
CYCLOPHOSPHAMIDE TABLETS IP 2007
dryness. Reserve a portion of the residue for identification combined filtrate and washings to dryness and dissolve the
test C. Dissolve 0.1 g in 10 ml of water and add 5 ml of silver residue in 50 ml of a 0.1 per cent w/v solution of sodium
nitrate solution; no precipitate is produced. Boil; a white hydroxide in 1,2-ethanediol. Boil the solution under a reflux
precipitate is produced which is insoluble in dilute nitric acid condenser for 30 minutes, allow to cool and rinse the
but is soluble in dilute ammonia solution from which it can be condenser with 25 ml of water. Add 75 ml of 2-propanol, 15 ml
reprecipitated by the addition of dilute nitric acid. of 2 M nitric acid, 10 ml of 0.1 M silver nitrate and 2 ml of
ferric ammonium sulphate solution and titrate with 0.1 M
C. Dissolve 0.1 g of the residue from test B in 3 ml of nitric
ammonium thiocyanate.
acid and 1 ml of sulphuric acid, heat till brown fumes are
evolved and the solution becomes colourless. Cool, add 10 ml 1 ml of 0.1 M silver nitrate is equivalent to 0.01305 g of
of water, heat again up to 60° and add 10 ml of ammonium C7H15Cl2N2O2P.
molybdate solution; a bright yellow precipitate is slowly Storage. Store in a refrigerator (2° to 8°). Avoid long exposure
formed. to temperatures above 30°. The solution should be used
immediately after preparation as it deteriorates on storage.
Tests
Labelling. The label states (1) the quantity of
pH (2.4.24). 4.0 to 6.0, determined in a 2.0 per cent w/v solution Cyclophosphamide in terms of the equivalent amount of
immediately after preparation. anhydrous cyclophosphamide; (2) the volume of Water for
Injections to be added; (3) that the solution should be used
Related substances. Determine by thin-layer chromatography immediately after preparation.
Mobile phase. A mixture of 80 volumes of 2-butanone,
12 volumes of water, 4 volumes of acetone and 2 volumes of
anhydrous formic acid. Cyclophosphamide Tablets
Test solution. Dissolve a quantity of the contents of the sealed Cyclophosphamide Tablets contain not less than 92.5 per cent
container containing 0.2 g of anhydrous cyclophosphamide and not more than 107.5 per cent of the stated amount of
in sufficient ethanol (95 per cent) to produce 10 ml and filter. anhydrous cyclophosphamide, C7H15Cl2N2O2P. The tablets are
Reference solution. Dilute 1 volume of solution (1) to coated.
100 volumes with ethanol (95 per cent).
Identification
dry the plate in a current of warm air and heat at 110° for A. Shake a quantity of the powdered tablets containing 0.2 g
10 minutes. Place the plate while hot in a tank in which is of anhydrous cyclophosphamide with 2 ml of chloroform and
placed a dish containing equal volumes of a 5 per cent w/v filter. The resulting solution complies with the following test.
solution of potassium permanganate and hydrochloric acid, Determine by infrared absorption spectrophotometry (2.4.6).
close the tank and allow to stand for 2 minutes. Remove the Compare the spectrum with that obtained with
plate and place it in a current of cold air until excess chlorine is cyclophosphamide RS or with the reference spectrum of
removed and an area of coating below the line of application cyclophosphamide.
gives not more than a faint blue colour with potassium iodide
B. Extract a quantity of the powdered tablets containing 0.25 g
of anhydrous cyclophosphamide with ether and evaporate
the extract to dryness. Preserve a portion of the residue for
identification test C. Dissolve 0.1 g in 10 ml of water and add
5 ml of silver nitrate solution; no precipitate is produced.
spot in the chromatogram obtained with the reference solution.
Boil; a white precipitate is produced which is insoluble in
Ignore any spot remaining on the line of application.
dilute nitric acid but is soluble in dilute ammonia solution
Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin from which it can be reprecipitated by the addition of dilute
Unit per mg of cyclophosphamide. nitric acid.
Assay. Determine the weight of the contents of 10 containers. C. Dissolve 0.1 g of the residue from test B in 3 ml of nitric acid
Shake vigorously a quantity of the mixed contents of the and 1 ml of sulphuric acid, heat till brown fumes are evolved
10 containers containing about 0.1 g of anhydrous and the solution becomes colourless. Cool, add 10 ml of water,
cyclophosphamide in 30 ml of chloroform for 15 minutes, filter heat again up to 60° and add 10 ml of ammonium molybdate
and wash the filter with 15 ml of chloroform. Evaporate the solution; a bright yellow precipitate is slowly formed.
362
IP 2007 CYCLOPROPANE
Tests minutes. Cool, add 1 ml of 0.1 M sodium acetate and mix. Add
1.6 ml of a 0.75 per cent w/v solution of 4-(4-nitrobenzyl)
Acidity. Shake a quantity of the powdered tablets containing pyridine in 1,2-ethanediol, mix and heat on a water-bath for
0.25 g of anhydrous cyclophosphamide with 20 ml of carbon 10 minutes. Cool, add 8.0 ml of a 2 per cent w/v solution of
dioxide-free water, filter and titrate the filtrate with 0.1 M sodium hydroxide in ethanol (95 per cent). Measure the
sodium hydroxide using phenolphthalein solution as absorbances of the solutions against the blank within 4
indicator; not more than 0.2 ml of 0.1 M sodium hydroxide is minutes at the maximum at about 560 nm (2.4.7).
required to change the colour of the solution.
Calculate the content of C7H15Cl2N2O2P in the tablet.
(2.4.17), coating the plate with silica gel G. Other tests. Comply with the tests stated under Tablets.
Mobile phase. A mixture of 80 volumes of 2-butanone, Assay. Weigh and powder 20 tablets. To an accurately weighed
12 volumes of water, 4 volumes of acetone and 2 volumes of quantity of the powder containing about 0.1 g of anhydrous
anhydrous formic acid. cyclophosphamide add 30 ml of chloroform, shake vigorously
for 15 minutes, filter and wash the filter with 15 ml of chloroform.
Test solution. Shake vigorously a quantity of the powdered Evaporate the combined filtrate and washings to dryness and
tablets containing 0.2 g of anhydrous cyclophosphamide with dissolve the residue in 50 ml of a 0.1 per cent w/v solution of
50 ml of chloroform for 15 minutes, filter, evaporate the filtrate sodium hydroxide in 1,2-ethanediol. Boil the solution under
to dryness and dissolve the residue in 10 ml of ethanol a reflux condenser for 30 minutes, allow to cool and rinse the
(95 per cent). condenser with 25 ml of water. Add 75 ml of 2-propanol, 15 ml
Reference solution. Dilute 1 volume of solution (1) to of 2 M nitric acid, 10 ml of 0.1 M silver nitrate and 2 ml of
100 volumes with ethanol (95 per cent). ferric ammonium sulphate solution and titrate with 0.1 M
ammonium thiocyanate.
dry the plate in a current of warm air and heat at 110° for 1 ml of 0.1 M silver nitrate is equivalent to 0.01305 g of
10 minutes. Place the plate while hot in a tank in which is C7H15Cl2N2O2P.
placed a dish containing equal volumes of a 5 per cent w/v Storage. Store at a temperature not exceeding 30°.
solution of potassium permanganate and hydrochloric acid,
close the tank and allow to stand for 2 minutes. Remove the Labelling. The label states the strength in terms of the
plate and place it in a current of cold air until excess chlorine is equivalent amount of anhydrous cyclophosphamide.
removed and an area of coating below the line of application
gives not more than a faint blue colour with potassium iodide
Cyclopropane
spot in the chromatogram obtained with the reference solution. H2
Ignore any spot remaining on the line of application. C
H2C CH2
Disintegration (2.5.1). Not more than 30 minutes.
Uniformity of content (for tablets containing 10 mg or less). C3H6 Mol. Wt. 42.1
Comply with the test stated under Tablets. Cyclopropane contains not less than 99.0 per cent v/v of C3H6.
Place one tablet in a 10-ml volumetric flask, add about 7 ml of Description. A colourless gas at atmospheric temperature and
water, shake until the tablet is completely disintegrated, dilute pressure; odour, characteristic; flammable.
with water to volume and filter. Wash the filter quantitatively
with 10 ml of water and combine the filtrate and washings (test NOTE — Mixtures of cyclopropane with oxygen or air at
solution). In another volumetric flask dissolve an accurately certain concentrations are explosive.
weighed quantity of cyclophosphamide RS in water to obtain
Tests
a solution of known concentration of about 500 µg per ml
(reference solution). Place in separate test-tubes (170 mm x Acidity or alkalinity. Dilute 0.3 ml of methyl red solution with
25 mm) 2.0 ml of the test solution, 2.0 ml of the reference 400 ml of boiling water and boil the solution for 5 minutes.
solution and 2.0 ml of water as the blank. Treat each tube in Cool to about 80° and pour 100 ml of solution into each of
the following manner. Add 0.7 ml of a 2.35 per cent v/v solution three matched Nessler cylinders marked A, B and C. To cylinder
of perchloric acid in water, mix and heat on a water-bath for 10 B add 0.2 ml of 0.01 M hydrochloric acid and to cylinder C
363
CYCLOSERINE IP 2007
add 0.4 ml of 0.01 M hydrochloric acid. Stopper both the and acidified in the same manner, add 7.5 ml of 0.001 M
cylinders and cool to room temperature. Pass a volume of the potassium bromide (standard solution). Transfer the solutions
gas equivalent to 2000 ml, measured at normal temperature to 100-ml matched Nessler cylinders, add 1.0 ml of 0.1 M silver
and pressure, through the solution in cylinder B, the time nitrate to each, dilute to 100 ml with water, mix well and allow
occupied being about 30 minutes. The colour of the solution to stand in the dark for 15 minutes. Compare the turbidities of
in cylinder B is not deeper red than that of the solution in the two solutions by viewing them both transversely and
cylinder C and not deeper yellow than that of the solution in vertically against a black background. The turbidity of the
cylinder A. test solution is not more intense than that of the standard
solution.
Carbon dioxide. Pass a volume of the gas equivalent to
1000 ml at normal temperature and pressure at a rate not Foreign odour. Transfer 10 ml of the material liquefied under
exceeding 4000 ml per hour through 100 ml of a 3 per cent w/v pressure to a cylinder cooled to a temperature not exceeding
solution of barium hydroxide contained in a vessel such that 40°, pour in successive small quantities onto a clean filter
the depth of the solution is between 12 and 14 cm, using a paper and allow it to evaporate spontaneously. No foreign
delivery tube having a bore of about 1 mm and extending to odour is detectable at any stage of the evaporation.
within 2 mm of the bottom of the vessel. The turbidity produced Assay. In a suitable nitrometer containing mercury, place a
is not more intense than that produced by adding 1 ml of a volume of the material liquefied under pressure equivalent to
solution of 0.1 g of sodium bicarbonate in 100 ml of carbon 80 to 100 ml of the gas, measured at normal temperature and
dioxide-free water to 100 ml of a 3 per cent w/v solution of pressure, add 25 ml of sulphuric acid and allow to stand for
barium hydroxide. 15 minutes. Not less than 99.0 per cent of its volume is
Ethanol and Water. Pass a volume of the gas equivalent to absorbed.
1000 ml, measured at normal temperature and pressure, through Storage. Store under pressure in metal cylinders in a cool
a weighed tube containing potassium hydroxide in small place.
pieces, the time occupied being 40 to 60 minutes. The increase
in weight of the tube is not more than 0.0056 g (equivalent to Labelling. The metal cylinder is painted orange and on the
0.3 per cent w/w of the Cyclopropane used). shoulder is stencilled the name of the gas or the symbol C3H6.
Unsaturated substances. Pass the gas coming out of the tube

of potassium hydroxide in the test for Ethanol and water
through a gas washing trap provided with a sintered-glass Cycloserine
bubbler containing 20.0 ml of iodine monochloride solution
and connected in series with two gas washing bottles O
NH
containing, respectively, 5.0 ml of iodine monochloride
solution and 10 ml of potassium iodide solution. Mix the
H2N O
contents of the trap and washing bottles and titrate with
0.1 M sodium thiosulphate. Add 10 ml of potassium iodide C3H6N2O2 Mol. Wt. 102.1
solution to 25.0 ml of iodine monochloride solution and titrate
with 0.1 M sodium thiosulphate. The difference between the Cycloserine is (R)-4-aminoisoxazolidin-3-one, an antimicrobial
titrations is not more than 1.8 ml (equivalent to 0.2 per cent substance produced by the growth of certain strains of
w/w of unsaturated substances, calculated as propylene). Streptomyces orchidaceous or S. garyphalus or obtained by
synthesis.
Halogen-containing substances. Pass a volume of the gas
equivalent to 1000 ml, measured at normal temperature and Cycloserine contains not less than 98.0 per cent and not more
pressure, with the necessary amount of air into a small mixing than 100.5 per cent of C3H6N2O2, calculated on the dried basis.
chamber and pass the resulting mixture through a heated quartz Description. A white or pale yellow, crystalline powder;
tube containing pieces of platinised quartz or through a heated hygroscopic.
silica tube containing sintered silica plates or pieces of
platinised quartz, the time occupied being not less than Identification
40 minutes. Absorb the products of combustion in 50 ml of a
3 per cent w/v solution of sodium peroxide. Boil the solution
for about 10 minutes, cool, neutralise with a solution of nitric
acid (containing about 30 per cent w/w of HNO3) and add 5 ml
of 2 M nitric acid (test solution). To 50 ml of the same solution B. To 1 ml of a 0.01 per cent w/v solution in 0.1 M sodium
of sodium peroxide which has been boiled, cooled, neutralised hydroxide add 3 ml of 1 M acetic acid and 1 ml of a freshly
364
IP 2007 CYCLOSERINE CAPSULES
prepared mixture of equal volumes of a 4 per cent w/v solution Identification

of sodium nitroprusside and 5 M sodium hydroxide; a blue
colour is produced slowly. A. Shake a quantity of the contents of the capsules containing
10 mg of Cycloserine with 100 ml of 1 M sodium hydroxide
Tests and filter. To 1 ml of the filtrate add 3 ml of 1 M acetic acid and
1 ml of a freshly prepared mixture of equal volumes of a 4 per
pH (2.4.24). 5.5 to 6.5, determined in a 10.0 per cent w/v solution. cent w/v solution of sodium nitroprusside and 5 M sodium
Specific optical rotation (2.4.22). +108° to +114°, determined hydroxide; a blue colour is produced slowly.
in a 5.0 per cent w/v solution in 2 M sodium hydroxide. B. In the Assay, the principal peak in the chromatogram
Heavy metals (2.3.13). 2.0 g complies with limit test for heavy obtained with the test solution corresponds to the peak in the
metals, Method B (10 ppm). chromatogram obtained with the reference solution.
Condensation products. Absorbance of a 0.04 per cent w/v
solution in 0.1 M sodium hydroxide at about 285 nm, not more
Tests
than 0.32 (2.4.7). Dissolution (2.5.2).
Sulphated ash (2.3.18). Not more than 0.5 per cent. Apparatus. No 2
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Medium. 900 ml of phosphate buffer pH 6.8.
on 1.0 g by drying in an oven at 60° at a pressure not exceeding Speed and time. 100 rpm and 30 minutes.
0.7 kPa for 3 hours.
Assay. Determine by liquid chromatography (2.4.14)
Test solution. Dissolve about 10 mg of the substance under
examination in 20.0 ml of the mobile phase. Dilute 5.0 ml of this Test solution. The filtrate diluted to produce a 0.028 per cent
solution to 50.0 ml with the mobile phase. w/v solution.
Reference solution. Dissolve 10 mg of the cycloserine RS in Reference solution. A 0.028 per cent w/v solution of cycloserine
20.0 ml of the mobile phase. Dilute 5.0 ml of this solution to RS in the dissolution medium.
50.0 ml with the mobile phase. Use the chromatographic system described under Assay.
Chromatographic system Calculate the content of C3H6N2O2.
octylsilyl silica gel (5 µm) (such as Wakosil C8 RS), D. Not less than 80 per cent of the stated amount of C3H6N2O2.
– mobile phase: 0.1 per cent w/v of methane sulphonic Condensation products. Weigh the contents of the capsules
acid and 0.78 per cent w/v of sodium dihydrogen containing about 0.5 g of Cycloserine, dissolve in 250 ml of
orthophosphate in water, the pH adjusted to 6.0 with 0.1 M sodium hydroxide solution, disperse with the aid of
dilute sodium hydroxide and filtered, ultrasound for 5 minutes. Dilute 5 ml of this solution to 25 ml
– flow rate. 1 ml per minute, with 0.1 M sodium hydroxide. Absorbance of the resulting
– spectrophotometer set at 227 nm, solution at about 285 nm, not more than 0.32 (2.4.7).
Inject the reference solution. The test is not valid unless the on 1.0 g of the contents of the capsules, by drying in an oven
relative standard deviation for replicate injections is not more at 60° at a pressure not exceeding 0.7 kPa for 3 hours.
than 2.0 per cent.
Calculate the content of C3H6N2O2.
Storage. Store protected from moisture Test solution. Mix the contents of 20 capsules. Weigh
accurately a quantity of the mixed contents of the capsules
containing about 250.0 mg of Cycloserine dissolve in
phosphate buffer pH 6.8, dilute to 250.0 ml with the same
Cycloserine Capsules solvent and filter. Dilute 5.0 ml of the filtrate to 25.0 ml with
phosphate buffer pH 6.8.
Cycloserine Capsules contain not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of Reference solution. A 0.02 per cent w/v solution of cycloserine
cycloserine, C3H6N2O2. RS in phosphate buffer pH 6.8.
365
CYCLOSERINE TABLETS IP 2007
Chromatographic system Assay. Weigh and powder 20 tablets. Weigh accurately a

– a stainless steel column 25 cm x 4.6 mm, packed with quantity of the powder containing about 0.1 g of Cycloserine,
octylsilyl silica gel (5 µm) ( such as Warkosil C8 RS), shake with 150 ml of water for 30 minutes, add sufficient water
– mobile phase: dissolve 1.0 g of methane sulphonic acid to produce 200.0 ml and filter. To 10.0 ml of the filtrate add
and 7.8 g of sodium dihydrogen orthophosphate 10 ml of water and 25 ml of 0.2 M sodium hydroxide, dilute to
dihydrate in 1000 ml of water and adjust pH to 6.0 with 50.0 ml with water and mix. To 4.0 ml of the mixture add 10 ml of
dilute sodium hydroxide, filter, 1 M acetic acid and 4 ml of sodium nitroprusside solution,
– flow rate. 1 ml per minute, dilute to 20 ml with 1 M acetic acid, mix and allow to stand for
– spectrophotometer set at 227 nm, 15 minutes. Measure the absorbance of the resulting solution
– a 20 µl loop injector. at the maximum at about 625 nm (2.4.7), using as the blank a
Inject the reference solution. The test is not valid unless the solution prepared by treating 4.0 ml of 0.1 M sodium hydroxide
tailing factor is not more than 2.0 and the relative standard in the same manner beginning at the words “add 10 ml of 1 M
deviation for replicate injections is not more than 2.0 per cent. acetic acid...” Calculate the content of C3H6N2O2 from the
absorbance obtained by repeating the operation using
Inject alternately the test solution and the reference solution. cycloserine RS in place of the powdered tablets.
Calculate the content of C3H6N2O2 in the capsules. Storage. Store at a temperature not exceeding 30°.
Cyproheptadine Hydrochloride
Cycloserine Tablets
Cycloserine Tablets contain not less than 92.5 per cent and
cycloserine, C3H6N2O2. ,HCl
Identification
A. Shake a quantity of the powdered tablets containing 0.5 g N
of Cycloserine with 25 ml of 1 M sodium hydroxide for
CH3
5 minutes and filter. The optical rotation of the filtrate is about
+2.2° (2.4.22).
C21H21N, HCl,1½H2O Mol. Wt. 332.9
B. To 0.2 ml of the filtrate obtained in test A add 3 ml of 1 M
Cyproheptadine Hydrochloride is 4-(5H-dibenzo[a,d]-
acetic acid and 1 ml of a freshly prepared mixture of equal
cyclohepten-5-ylidene)-1-methylpiperidine hydrochloride.
volumes of a 4 per cent w/v solution of sodium nitroprusside
and 5 M sodium hydroxide; a blue colour is produced slowly. Cyproheptadine Hydrochloride contains not less than
98.5 per cent and not more than 101.0 per cent of C21H21N,HCl,
Tests calculated on the dried basis.
Light absorption. Shake a quantity of the powdered tablets Description. A white or slightly yellow, crystalline powder.
containing 0.25 g of Cycloserine with 80 ml of 0.1 M sodium
hydroxide for 10 minutes, add sufficient 0.1 M sodium Identification
hydroxide to produce 100.0 ml, mix and filter. Dilute a suitable
volume of the filtrate with sufficient 0.1 M hydrochloric acid
B and C may be omitted if tests A and D are carried out.
to produce a solution containing 0.0025 per cent w/v of
Cycloserine. Absorbance of the resulting solution, measured A. Dissolve 0.1 g in 10 ml of water, make alkaline with 1 M
within 15 minutes of preparing the final solution, at the maximum sodium hydroxide, extract with 5 ml of dichloromethane, dry
at about 219 nm, 0.78 to 0.96 (2.4.7). over anhydrous sodium sulphate and remove the solvent with
the aid of a current of nitrogen. The oily residue complies with
Disintegration (2.5.1). Not more than 30 minutes.
the following test.
on 1.0 g of the powdered tablets, by drying in an oven at
Compare the spectrum with that obtained with cyproheptadine
about 60° at a pressure not exceeding 0.7 kPa for 3 hours.
Other tests. Comply with the tests stated under Tablets. reference spectrum of cyproheptadine.
366
IP 2007 CYPROHEPTADINE SYRUP
B. When examined in the range 230 nm to 360 nm, a 0.002 per Sulphated ash (2.3.18). Not more than 0.1 per cent.
cent w/v solution in ethanol (95 per cent) shows an absorption Loss on drying (2.4.19). 7.0 to 9.0 per cent, determined on
maximum only at about 286 nm; absorbance at about 286 nm, 1.0 g by drying in an oven at 100° at a pressure not exceeding
about 0.67 (2.4.7). 0.7 kPa.
C. Determine by thin-layer chromatography (2.4.17) coating
acetic anhydride and 20 ml of anhydrous glacial acetic acid
Mobile phase. A mixture of 75 volumes of cyclohexane, and add 10 ml of mercuric acetate solution. Titrate with 0.1 M
20 volumes of ether and 5 volumes of diethylamine. perchloric acid, using crystal violet solution as indicator.
Test solution. Dissolve 0.1 g of the substance under Carry out a blank titration.
examination in 100 ml of methanol. 1 ml of 0.1 M perchloric acid is equivalent to 0.03239 g of
Reference solution (a). A 0.1 per cent w/v solution of C21H21N,HCl.
cyproheptadine hydrochloride RS in methanol. Storage. Store protected from light.
w/v of each of imipramine hydrochloride RS and
cyproheptadine hydrochloride RS in methanol.
Apply to the plate 2 µl of each solution. After development, Cyproheptadine Syrup
Cyproheptadine Hydrochloride Syrup
solution corresponds to that in the chromatogram obtained Cyproheptadine Syrup contains not less than 90.0 per cent
with reference solution (a). The test is not valid unless the and not more than 110.0 per cent of the stated amount of
chromatogram obtained with reference solution (b) shows two cyproheptadine hydrochloride, C21H21N,HCl.
clearly separated principal spots.
Identification
D. A saturated solution gives reaction A of chlorides (2.3.1).
To 5 ml add 5 ml of a 1 per cent w/v solution of sodium
Tests
bicarbonate and extract with three quantities, each of 15 ml,
Related substances. Determine by thin-layer chromatography of 2,2,4-trimethylpentane. Wash the combined 2,2,4-
(2.4.17) coating the plate with silica gel GF254. trimethylpentane extracts with 5 ml of the sodium bicarbonate
solution and discard the washings. Evaporate the 2,2,4-
Mobile phase. A mixture of 90 volumes of dichloromethane
trimethylpentane solution to dryness on a water-bath and
and 10 volumes of methanol.
dissolve the residue in 100 ml of ethanol (95 per cent). When
Solvent mixture. A mixture of 9 volumes of dichloromethane examined in the range 230 nm to 360 nm, the resulting solution
and 1 volumes of methanol. shows an absorption maximum only at about 286 nm (2.4.7).
examination in 10 ml with solvent mixture. Tests
Reference solution (a). A 0.001 per cent w/v solution of the pH (2.4.24). 3.5 to 4.5.
substance under examination in the same solvent mixture. Other tests. Comply with the tests stated under Oral Liquids.
Reference solution (b). A solution containing 0.002 per cent Assay. To an accurately measured volume of the syrup
w/v of dibenzocycloheptatriene RS in the same solvent containing about 2 mg of Cyproheptadine Hydrochloride add
mixture. 20 ml of a 1 per cent w/v solution of sodium bicarbonate and
Apply to the plate 10 µl of each solution. After development, extract with two quantities, each of 25 ml, of 2,2,4-
dry the plate in air and spray with ethanolic sulphuric acid trimethylpentane. Wash the combined 2,2,4-trimethylpentane
(20 per cent), heat at 110° for 30 minutes. Allow to cool and extracts with 5 ml of the sodium bicarbonate solution and
examine in ultraviolet light at 365 nm. Any spot corresponding discard the washings. Extract the 2,2,4-trimethylpentane
to dibenzocycloheptatriene in the chromatogram obtained with solution with 50 ml of 0.05 M sulphuric acid and collect the
the test solution is not more intense than the spot in the aqueous extract in a 100-ml volumetric flask. Dilute to volume
chromatogram obtained with reference solution (b) and any with 0.05 M sulphuric acid and mix. Filter a portion of the
other secondary spot in the chromatogram obtained with the solution through a dry filter paper and discard the first 20 ml
test solution is not more intense than the spot in the of the filtrate. Measure the absorbance of the filtrate at the
chromatogram obtained with reference solution (a). maximum at about 286 nm (2.4.7), using 0.05 M sulphuric acid
367
CYPROHEPTADINE TABLETS IP 2007
as the blank. Calculate the content of C21H21N,HCl taking 355 Apply to the plate 10 µl of each solution. After development,
as the specific absorbance at 286 nm. dry the plate in air and spray with ethanolic sulphuric acid
(20 per cent). Heat at 110° for 30 minutes and examine in
ultraviolet light at 365 nm. In the chromatogram obtained with
test solution (a) any spot corresponding to
Cyproheptadine Tablets dibenzocycloheptatriene is not more intense than the spot in
Cyproheptadine Hydrochloride Tablets the chromatogram obtained with reference solution (c) and
any other secondary spot is not more intense than the spot in
Cyproheptadine Tablets contain not less than 90.0 per cent the chromatogram obtained with reference solution (a).
cyproheptadine hydrochloride, C21H21N,HCl. Uniformity of content. Comply with the test stated under
Tablets.
Identification Powder one tablet, warm with 20 ml of ethanol (95 per cent)
A. To a quantity of the powdered tablets containing 20 mg of and centrifuge. Repeat the extraction with three further
Cyproheptadine Hydrochloride add 10 ml of water and 2.5 ml quantities, each of 20 ml, of ethanol (95 per cent). Cool the
of 0.1 M sodium hydroxide, extract with 10 ml of combined extracts and add sufficient ethanol (95 per cent) to
dichloromethane, filter through anhydrous sodium sulphate produce 200.0 ml. Measure the absorbance of the resulting
placed over absorbent cotton moistened with solution at the maximum at about 286 nm (2.4.7). Calculate the
dichloromethane and evaporate the filtrate to dryness. The content of C21H21N,HCl taking 355 as the specific absorbance
residue complies with the following test. at 286 nm.
Determine by infrared absorption spectrophotometry (2.4.6). Other tests. Comply with the tests stated under Tablets.
Compare the spectrum with that obtained with cyproheptadine Assay. Weigh and powder 20 tablets. Weigh accurately a
hydrochloride RS treated in the same manner or with the quantity of the powder containing about 1.5 mg of
reference spectrum of cyproheptadine. Cyproheptadine Hydrochloride, add sufficient ethanol
B. In the test for Related substances, the principal spot in the (95 per cent) to produce 100.0 ml, mix well and filter. Measure
chromatogram obtained with test solution (b) corresponds to the absorbance of the filtrate at the maximum at about 286 nm
that in the chromatogram obtained with reference solution (b). (2.4.7). Calculate the content of C21H21N,HCl taking 355 as the
C. Extract a quantity of the powdered tablets containing 20 mg
of Cyproheptadine Hydrochloride with 7 ml of water, filter,
add 0.3 ml of 5 M ammonia to the filtrate and filter again. The
filtrate gives reaction A of chlorides (2.3.1).
Cytarabine
Tests
β-Cytosine Arabinoside
(2.4.17), using a precoated silica gel plate.
Mobile phase. A mixture of 90 volumes of dichloromethane NH2
and 10 volumes of methanol. N
Test solution (a). Shake mechanically for 10 minutes a quantity
of the powdered tablets containing 50 mg of Cyproheptadine HO O N
Hydrochloride with 5 ml of the mobile phase and filter (such O
as Whatman GF/C filter paper). HO
Test solution (b). Dilute 1 volume of test solution (a) to OH

10 volumes with the mobile phase.
Reference solution (a). Dilute 1 volume of test solution (a)
C9H13N3O5 Mol. Wt. 243.2
serially in two steps to 1000 volumes with the mobile phase.
Cytarabine is 1-β-D-arabinofuranosylcytosine.
w/v of cyproheptadine hydrochloride RS in the mobile phase. Cytarabine contains not less than 99.0 per cent and not more
Reference solution (c). A solution containing 0.002 per cent than 100.5 per cent of C9H13N3O5, calculated on the dried basis.
w/v of dibenzocycloheptatriene RS in the mobile phase. Description. A white or almost white, crystalline powder.
368
IP 2007 CYTARABINE INJECTION
CAUTION — Cytarabine is very poisonous. Great care should Cytarabine intended for use in the manufacture of parenteral
be taken to avoid inhaling the particles of cytarabine and preparations without a further appropriate procedure for
exposing the skin to the dried substance. the removal of bacterial endotoxins complies with the
following additional requirement.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). Unit per mg.
Compare the spectrum with that obtained with cytarabine RS Cytarabine intended for use in the manufacture of parenteral
or with the reference spectrum of cytarabine. preparations without a further appropriate sterilisation
B. When examined in the range 230 nm to 360 nm, a 0.001 per procedure complies with the following additional
cent w/v solution in 0.1 M hydrochloric acid shows an requirement.
absorption maximum only at about 280 nm; absorbance at Sterility. Complies with the test for sterility (2.2.11).
about 280 nm, about 0.55 (2.4.7).
Storage. Store protected from light. If it is intended for use in
C. In the test for Related substances, the principal spot in the the manufacture of parenteral preparations, the container
chromatogram obtained with test solution (b) corresponds to should be sterile, tamper-evident and sealed so as to exclude
that in the chromatogram obtained with reference solution micro-organisms.
(b).
Labeling. The label states whether or not the material is
Tests intended for use in the manufacture of parenteral preparations.
Cytarabine Injection
(2.4.17), coating the plate with silica gel GF254. β-Cytosine Arabinoside Injection
Mobile phase. A mixture of 65 volumes of 2-butanone, Cytarabine for Injection is a sterile material consisting of
20 volumes of acetone and 15 volumes of water. Cytarabine with or without excipients. It is filled in a sealed
Test solution (a). A 5 per cent w/v solution of the substance container.
under examination in water. The injection is constituted by dissolving the contents of the
Test solution (b). A 0.02 per cent w/v solution of the substance sealed container in the requisite amount of sterile Water for
under examination in water. Injections, immediately before use.
Reference solution (a). A 0.0025 per cent w/v solution of the The constituted solution complies with the requirements for
substance under examination in water . Clarity of solution and Particulate matter stated under
Storage. The constituted solution should be used immediately
w/v solution of cytarabine RS in water.
Apply to the plate 5 µl of each solution. After development, recommended by the manufacturer.
Cytarabine Injection contains not less than 95.0 per cent and
Any secondary spot in the chromatogram obtained with test
solution (a) is not more intense than the spot in the
cytarabine, C9H13N3O5.
60° for 3 hours at a pressure of 0.2 kPa to 0.7 kPa.
Assay. Weigh accurately about 0.5 g and dissolve in 40 ml of Identification
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Mix 0.1 g of the substance under examination with 10 ml of hot
acid, using 1-naphtholbenzein solution as indicator. Carry ethanol (95 per cent), filter, allow the filtrate to cool and induce
out a blank titration. crystallisation if necessary. Filter, wash the crystals with 2 ml
1 ml of 0.1 M perchloric acid is equivalent to 0.02432 g of of ethanol (95 per cent) and dry at 60° at a pressure of 0.7
C9H13N3O5. kPa. The residue complies with the following test.
369
CYTARABINE INJECTION IP 2007
Determine by infrared absorption spectrophotometry (2.4.6). with an Rf value of about 1.1 relative to the spot in the
Compare the spectrum with that obtained with cytarabine RS chromatogram obtained with reference solution (b) is not more
or with the reference spectrum of cytarabine. intense than the spot in the chromatogram obtained with
reference solution (b). Any other secondary spot in the
Tests chromatogram obtained with the test solution is not more
reference solution (a).
in the solvent stated on the label.
0.8 g.
unit per mg.
20 volumes of acetone and 15 volumes of water.
Weigh accurately about 0.5 g of the mixed contents of the
under examination in water.
10 containers and dissolve by heating, if necessary, in 80 ml of
Reference solution (a). A 0.02 per cent w/v solution of the anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
substance under examination water. acid, using 1-naphtholbenzein solution as indicator. Carry
Reference solution (b). A solution containing 0.04 per cent out a blank titration.
w/v solution of uridine water. 1 ml of 0.1 M perchloric acid is equivalent to 0.02432 g of
Apply to the plate 5 µl of each solution. After development, C9H13N3O5.
dry the plate in air and examine in ultraviolet light at 254 nm. Storage. Store protected from light.
Any spot in the chromatogram obtained with the test solution
370
D
Danazol ....
Danazol Capsules ....
Dapsone ....
Dapsone Tablets ....
Dehydroacetic Acid ....
Dehydroemetine Hydrochloride ....
Dehydroemetine Injection ....
Dequalinium Chloride ....
Desferrioxamine Mesylate ....
Desferrioxamine Injection ....
Deslanoside ....
Deslanoside Injection ....
Desoxycortone Acetate ....
Desoxycortone Acetate Injection ....
Dexamethasone ....
Dexamethasone Tablets ....
Dexamethasone Sodium Phosphate ....
Dexamethasone Injection ....
Dextran 40 Injection ....
Dextrin ....
Dextromethorphan Hydrobromide ....
Dextromethorphan Hydrobromide Syrup ....
Dextrose ....
Dextrose Injection ....
Diazepam ....
Diazepam Capsules ....
Diazepam Injection ....
Diazepam Tablets ....
371
Dibutyl Phthalate ....

Diclofenac Sodium ....
Diclofenac Injection ....
Diclofenac Tablets ....
Dicyclomine Hydrochloride ....
Dicyclomine Injection ....
Dicyclomine Oral Solution ....
Dicyclomine Tablets ....
Didanosine ....
Didanosine Capsules ....
Didanosine Tablets ....
Dienoestrol ....
Dienoestrol Tablets ....
Diethylcarbamazine Citrate ....
Diethylcarbamazine Tablets ....
Diethyl Phenyl Acetamide ....
Diethyl Phthalate ....
Diethyltoluamide ....
Digitoxin ....
Digitoxin Tablets ....
Digoxin ....
Digoxin Injection ....
Digoxin Paediatric Solution ....
Digoxin Tablets ....
Diiodohydroxyquinoline ....
Diiodohydroxyquinoline Tablets ....
Diloxanide Furoate ....
Diloxanide Tablets ....
Diltiazem Hydrochloride ....
Diltiazem Tablets ....
Dimercaprol ....
Dimercaprol Injection
372
Activated Dimethicone ....

Diphenhydramine Hydrochloride ....
Diphenhydramine Capsules ....
Diphenoxylate Hydrochloride ....
Disodium Edetate ....
Disodium Edetate Injection ....
Disulfiram ....
Disulfiram Tablets ....
Dithranol ....
Dithranol Ointment ....
Docusate Sodium ....
Domperidone Maleate ....
Domperidone Tablets ....
Donepezil Hydrochloride ....
Donepezil Tablets ....
Dothiepin Hydrochloride ....
Dothiepin Capsules ....
Doxepin Hydrochlorude ....
Doxepin Capsules ....
Doxorubicin Hydrochloride ....
Doxorubicin Injection ....
Doxycycline Hydrochloride ....
Doxycycline Capsules ....
Dydrogesterone ....
Dydrogesterone Tablets ....
373
IP 2007 DANAZOL

Danazol compact spot.
H3C OH Tests
C CH
H3 C H Specific optical rotation (2.4.22). +21.0° to +27.0°, determined
N H H Related substances. Determine by thin-layer chromatography
O
C22H27NO2 Mol. Wt. 337.5 Mobile phase. A mixture of 70 volumes of cyclohexane and 30
Danazol is 17α-pregna-2,4-diene-20-yno[2,3-d]isoxazol-17-ol. volumes of ethyl acetate.
Danazol contains not less than 97.0 per cent and not more Test solution. Dissolve 0.5 g in a mixture of 9 volumes of
than 102.0 per cent of C22H27NO2, calculated on the dried basis. chloroform and 1 volume of methanol.
Description. A white to pale yellow, crystalline powder. Reference solution (a). Dissolve 50 mg of danazol RS in
100 ml of the same solvent mixture.
Identification Reference solution (b). Dilute 10 ml of reference solution (a)
A. Determine by infrared absorption spectrophotometry (2.4.6). to 20 ml with the same solvent mixture.
Compare the spectrum with that obtained with danazol RS or Apply to the plate 5 µl of each solution. After development,
with the reference spectrum of danazol. dry the plate in warm air and examine in ultraviolet light at
B. When examined in the range 230 nm to 360 nm (2.4.7), the 254 nm. Expose the plate to the vapour of iodine for 5 minutes
final solution obtained in the Assay shows an absorption and examine the plate again. By both methods of visualisation,
maximum only at about 285 nm. any secondary spot in the chromatogram obtained with the
C. Determine by thin-layer chromatography (2.4.17), coating chromatogram obtained with reference solution (a) and not
the plate with silica gel G. more than one such spot is more intense than the spot in the
Solvent mixture. A mixture of 90 volumes of chloroform and chromatogram obtained with reference solution (b).
10 volumes of methanol. Loss on drying (2.4.19). Not more than 2.0 per cent, determined
Mobile phase. A mixture of 70 volumes of cyclohexane and 30 on 1.0 g by drying in an oven at 60° at a pressure not exceeding
volumes of ethyl acetate. 2.7 kPa.
Test solution. Dissolve 25 mg of the substance under Assay. Weigh accurately about 0.1 g, previously dried, dissolve
examination in 10 ml of the solvent mixture. in 50 ml of ethanol (95 per cent), swirling until dissolved, and
dilute to 100.0 ml with ethanol (95 per cent). Dilute 2.0 ml of
Reference solution (a). Dissolve 25 mg of danazol RS in 10 ml
this solution to 100.0 ml with ethanol (95 per cent). Measure
of the solvent mixture.
Reference solution (b). Mix equal volumes of the test solution about 285 nm (2.4.7). Calculate the content of C22H27NO2 from
and reference solution (a). the absorbance obtained by repeating the procedure using a
Place the dry plate in a tank containing a shallow layer of the solution containing 0.002 per cent w/v of danazol RS in place
solvent mixture, allow the liquid to ascend to the top, remove of the substance under examination.
the plate from the tank and allow the solvent to evaporate. Storage. Store protected from light.
Use within 2 hours, with the flow of the mobile phase in the
direction in which the aforementioned treatment was done.
Apply to the plate 1 µl of each solution. Allow the mobile Danazol Capsules
phase to rise 12 cm. Dry the plate in a current of warm air, allow
the solvent to evaporate, heat at 120° for 15 minutes and spray Danazol Capsules contain not less than 90.0 per cent and not
the hot plate with ethanolic sulphuric acid (20 per cent v/v). more than 110.0 per cent of the stated amount of danazol,
Heat at 120° for a further 10 minutes, allow to cool and examine C22H27NO2.
Identification
corresponds to that in the chromatogram obtained with A. Extract the contents of the capsules containing about
reference solution (a). The principal spot in the chromatogram 50 mg of Danazol with 50 ml of chloroform, filter and evaporate
375
DAPSONE IP 2007
the filtrate to dryness on a water-bath in a stream of nitrogen. D. 2 ml of a 0.005 per cent w/v solution in 0.1 M hydrochloric
The residue complies with the following test. acid gives the reaction of primary aromatic amines (2.3.1).
Determine by infrared absorption spectrophotometry (2.4.6). Tests
Compare the spectrum with that obtained with danazol RS or
with the reference spectrum of danazol. Related substances. Determine by thin-layer chromatography
Tests
Mobile phase. A mixture of 20 volumes of n-heptane,
Other tests. Comply with the tests stated under Capsules. 20 volumes of ethyl acetate, 6 volumes of methanol and
Assay. Weigh accurately a quantity of the mixed contents of 1 volume of strong ammonia solution.
20 capsules containing about 0.1 g of Danazol, dissolve in Test solution (a). Dissolve 0.1 g of the substance under
50 ml of chloroform, shake well for 5 minutes, dilute to 100.0 ml examination in 10 ml of methanol.
with chloroform and filter. Dilute 2.0 ml of the filtrate to
100.0 ml with chloroform and measure the absorbance of the
methanol.
Calculate the content of C22H27NO2 from the absorbance Reference solution (a). Dilute 1 ml of test solution (b) to 10 ml
obtained by repeating the procedure using a solution with methanol.
containing 0.002 per cent w/v of danazol RS in place of the
Reference solution (c). A 0.1 per cent w/v solution of dapsone
RS in methanol.
Dapsone dry the plate in air, spray with a 0.1 per cent w/v solution of
O O 4-dimethylaminocinnamaldehyde in a mixture of 99 volumes
S of ethanol (95 per cent) and 1 volume of hydrochloric acid
and examine in daylight. Any secondary spot in the
chromatogram obtained with the test solution (a) is not more
H2N NH2 intense than the spot in the chromatogram obtained with
reference solution (a) and not more than two such spots are
C12H12N2O2S Mol. Wt. 248.3 more intense than the spot in the chromatogram obtained
Dapsone is the bis(4-aminophenyl)sulphone. with reference solution (b).
Dapsone contains not less than 99.0 per cent and not more Sulphated ash (2.3.18). Not more than 0.1 per cent.
than 101.0 per cent of C12H12N2O2S, calculated on the dried Loss on drying (2.4.19). Not more than 1.5 per cent, determined
basis. on 1.0 g by drying in an oven at 105°.
Description. A white or creamy-white, crystalline powder.
Identification of 20 ml of water and 20 ml of hydrochloric acid. Cool the
solution to about 15° and determine by the nitrite titration
Test A may be omitted if tests B, C and D are carried out. Tests (2.3.31). Carry out a blank titration.
1 ml of 0.1 M sodium nitrite is equivalent to 0.01242 g of
A. Determine by infrared absorption spectrophotometry (2.4.6). C12H12N2O2S.
Compare the spectrum with that obtained with dapsone RS or
with the reference spectrum of dapsone. Storage. Store protected from light.

maxima at about 260 nm and 295 nm; absorbance at about
260 nm, about 0.36 and at about 295 nm, about 0.6. Dapsone Tablets
C. In the test for Related substances, the principal spot in the Dapsone Tablets contain not less than 93.0 per cent and not
chromatogram obtained with test solution (b) corresponds to more than 107.0 per cent of the stated amount of dapsone,
that in the chromatogram obtained with reference solution (c). C12H12N2O2S.
376
IP 2007 DEHYDROACETIC ACID
Identification accurately measured volume of the filtrate containing about

0.2 mg of Dapsone to a 25-ml volumetric flask, add 5 ml of 1 M
A. Shake a quantity of the powdered tablets containing 0.1g sodium hydroxide, dilute to volume with water and mix.
of Dapsone with 10 ml of acetone, filter and evaporate the Measure the absorbance of the resulting solution at the
filtrate to dryness. The residue complies with the following maximum at about 290 nm (2.4.7). Calculate the content of
test. C12H12N2O2S from the absorbance obtained from a solution
Determine by infrared absorption spectrophotometry (2.4.6). prepared by adding 5 ml of 1 M sodium hydroxide to 20 ml of
Compare the spectrum with that obtained with dapsone RS or a 2 per cent v/v solution of hydrochloric acid containing
with the reference spectrum of dapsone. 0.2 mg of dapsone RS and adding sufficient water to produce
25.0 ml.
chromatogram obtained with test solution (b) corresponds to D. Not less than 75 per cent of the stated amount of
that in the chromatogram obtained with reference solution (c). C12H12N2O2S.
Tests
Related substances. Determine by thin-layer chromatography quantity of the powder containing about 0.25 g of Dapsone
(2.4.17), coating the plate with silica gel G. and dissolve in a mixture of 15 ml of water and 15 ml of 2 M
hydrochloric acid. Cool the solution to about 15° and
Mobile phase. A mixture of 20 volumes of n-heptane,
determine by the nitrite titration (2.3.31). Carry out a blank
20 volumes of ethyl acetate, 6 volumes of methanol and
titration.
1 volume of strong ammonia solution.
1 ml of 0.1 M sodium nitrite is equivalent to 0.01242 g of
Test solution (a). Shake a quantity of the powdered tablets
C12H12N2O2S.
containing 0.1 g of Dapsone with 10 ml of methanol and filter.
methanol.
with methanol. Dehydroacetic Acid
10 ml with methanol. H3 C O O H3C O O
Reference solution (c). A 0.1 per cent w/v solution of dapsone
O O
RS in methanol.
Apply to the plate 10 µl of each solution. After development, O CH3 OH CH3
dry the plate in air, spray with a 0.1 per cent w/v solution of
4-dimethylaminocinnamaldehyde in a mixture of 99 volumes C8H8O4 Mol. Wt. 168.1
of ethanol (95 per cent) and 1 volume of hydrochloric acid
and examine in daylight. Any secondary spot in the Dehydroacetic Acid is a tautomeric mixture of 3-acetyl-6-
chromatogram obtained with the test solution (a) is not more methyl-2H-pyran-2,4(3H)-dione and 3-acetyl-4-hydroxy-6-
intense than the spot in the chromatogram obtained with methyl-2H-pyran-2-one
reference solution (a) and not more than two such spots are Dehydroacetic Acid contains not less than 98.0 per cent and
more intense than the spot in the chromatogram obtained not more than 100.5 per cent of C8H8O4, calculated on the
with reference solution (b). anhydrous basis.
Dissolution (2.5.2). Description. A white or almost white, crystalline powder;
Apparatus. No 1 odourless or practically odourless.
Medium. 900 ml of a 2 per cent w/v solution of hydrochloric
acid.
Identification
Speed and time. 100 rpm and 60 minutes. A. Determine by infrared absorption spectrophotometry (2.4.6).
Withdraw a suitable volume of the medium and filter through Compare the spectrum with that obtained with dehydroacetic
a membrane filter disc with an average pore diameter not greater acid RS.
than 1.0 µm, rejecting the first few ml of the filtrate. Transfer an B. Melts at 109° to 111° (2.4.21).
377
DEHYDROEMETINE HYDROCHLORIDE IP 2007
Tests Description. A white to yellowish-white, crystalline powder;

odourless.
Arsenic (2.3.10). Heat gently 3.3 g with 2 ml of nitric acid and
0.5 ml of sulphuric acid in a long-necked flask until the first Identification
reaction has subsided, cool, add carefully and in small portions,
15 ml of nitric acid and 6 ml of sulphuric acid, taking care to A. When examined in the range 230 nm to 360 nm (2.4.7), a
avoid excessive foaming. Continue heating, adding further 0.005 per cent w/v solution in 0.1 M hydrochloric acid shows
small portions of nitric acid, if necessary, until white fumes an absorption maximum only at about 282 nm; absorbance at
are evolved and the solution becomes colourless or almost about 282 nm, about 0.62.
colourless. Cool, add carefully 10 ml of water, evaporate until B. Sprinkle 5 mg on the surface of a 5 per cent w/v solution of
white fumes are evolved. Repeat the addition of water and ammonium molybdate in sulphuric acid; a green colour
evaporation until all the nitric acid has been removed, cool, develops.
dilute to 50 ml with water and add 10 ml of stannated
C. Gives reaction A of chlorides (2.3.1).
hydrochloric acid AsT. The resulting solution complies with
the limit test for arsenic (3 ppm). Tests
Heavy metals (2.3.13). 2.0 g complies with the limit test for Appearance of solution. A 5.0 per cent w/v solution is clear
heavy metals, Method B (10 ppm). (2.4.1), and not more intensely coloured than reference solution
Sulphated ash (2.3.18). Not more than 0.1 per cent, determined YS5 or BYS6 (2.4.1).
on 2.0 g. pH (2.4.24). 3.5 to 5.0, determined in a 3.0 per cent w/v solution.
Water (2.3.43). Not more than 1.0 per cent, determined on Heavy metals (2.3.13). 1.0 g complies with the limit test for
2.0 g. heavy metals, Method B (20 ppm).
Assay. Weigh accurately about 0.5 g, dissolve in 75 ml of Sulphated ash (2.3.18). Not more than 0.1 per cent.
previously neutralised ethanol (95 per cent), add
phenolphthalein solution and titrate with 0.1 M sodium Loss on drying (2.4.19). Not more than 7.0 per cent, determined
hydroxide to a pink end-point that persists for not less than on 1.0 g by drying in an oven at 100° at a pressure not exceeding
30 seconds. 0.7 kPa for 4 hours.
1 ml of 0.1 M sodium hydroxide is equivalent to 0.01681 g of Assay. Weigh accurately about 0.4 g, dissolve in 40 ml of
C8H8O4. anhydrous glacial acetic acid and add 15 ml of mercuric
Dehydroemetine Hydrochloride C29H38N2O4,2HCl.
H3CO
N
H3CO
H
Dehydroemetine Injection
CH3
Dehydroemetine Hydrochloride Injection
,2HCl
Dehydroemetine Injection is a sterile solution of
OCH3
HN Dehydroemetine Hydrochloride in Water for Injections.
Dehydroemetine Injection contains not less than 95.0 per cent
OCH3
dehydroemetine hydrochloride, C29H38N2O4,2HCl.
C29H38N2O4,2HCl Mol. Wt. 551.6
Description. A clear, almost colourless solution.
Dehydroemetine Hydrochloride is 2,3-didehydro-6′,7′,10, 11-
tetramethoxyemetan dihydrochloride Identification
Dehydroemetine Hydrochloride contains not less than A. To a volume containing 30 mg of Dehydroemetine
98.5 per cent and not more than 101.5 per cent of Hydrochloride add 1 ml of 0.05 M iodine; a yellowish-brown
C29H38N2O4,2HCl, calculated on the dried basis. precipitate is produced.
378
IP 2007 DESFERRIOXAMINE MESYLATE
B. To a volume containing 15 mg of Dehydroemetine Tests

Hydrochloride add 1 ml of potassium mercuri-iodide solution;
a white precipitate is produced. Acidity or alkalinity. Shake 0.1 g for 10 minutes with 100 ml of
carbon dioxide-free water and add 0.5 ml of bromocresol
Tests purple solution. Not more than 0.2 ml of 0.1 M hydrochloric
acid or 0.1 M sodium hydroxide is required to change the
pH (2.4.24). 2.8 to 5.0. colour of the solution.
Other tests. Complies with the tests stated under Parenteral Non-quaternised amines. Not more than 1.0 per cent,
Preparations (Injections). calculated as 4-aminoquinaldine, C10H10N2, on the dried basis
Assay. To an accurately measured volume containing 60 mg and determined by the following method. Shake 1.0 g with
of Dehydroemetine Hydrochloride add sufficient 0.1 M 45 ml of water for 5 minutes, add 5 ml of dilute nitric acid and
hydrochloric acid to produce 100.0 ml. Dilute 5.0 ml to shake for 10 minutes. Filter through cotton wool. Transfer
100.0 ml with 0.1 M hydrochloric acid, mix and measure the 20.0 ml of the filtrate to a separator, add 20 ml of 1 M sodium
absorbance of the resulting solution at the maximum at about hydroxide, extract with two quantities, each of 50 ml, of ether,
282 nm (2.4.7). Calculate the content of C29H38N2O4,2HCl taking washing each extract in turn with the same 5 ml of water, and
123 as the specific absorbance at the maximum at about then extract each ether extract successively with 20 ml, 20 ml
282 nm. and 5 ml of 1 M hydrochloric acid. Combine the acid extracts,
dilute to 50.0 ml with 1 M hydrochloric acid and measure the
Storage. Store protected from light, in single dose container. absorbance of the resulting solution at the maximum at about
319 nm and 326.5 nm (2.4.7). The ratio of the absorbance at
319 nm to that at 326.5 nm is not less than 1.0. Calculate the
Dequalinium Chloride percentage of C10H10N2 from the expression 0.387a - 0.306b,
where a and b are the specific absorbances at about 319 nm
and 326.5 nm respectively.
H2N N (CH2)10 N NH2 2Cl
on 1.0 g by drying in an oven at 105° for 3 hours at a pressure
not exceeding 0.7 kPa.
CH3 H3C
Assay. Weigh accurately about 0.5 g, dissolve in a mixture of
80 ml of anhydrous glacial acetic acid and 20 ml of mercuric
C30H40Cl2N4 Mol. Wt. 527.7
Dequalinium Chloride is 4,4′-diamino-2,2′-dimethyl-N,N′- crystal violet solution as indicator. Carry out a blank titration.
decamethylenedi (quinolinium chloride). 1 ml of 0.1 M perchloric acid is equivalent to 0.02638 g of
Dequalinium Chloride contains not less than 95.0 per cent and C30H40Cl2N4.
not more than 101.0 per cent of C30H40Cl2N4, calculated on the
dried basis.
Description. A creamy white powder; odourless or almost
Desferrioxamine Mesylate
odourless. Deferoxamine Mesylate; Deferoxamine Mesilate
Identification OH O
H
H3C N
A. Determine by infrared absorption spectrophotometry (2.4.6). (CH2)5 N
Compare the spectrum with that obtained with dequalinium O O N OH ,CH3SO3H
chloride RS or with the reference spectrum of dequalinium O H
chloride. (CH2)5 N
H2N N (CH2)5
B. When examined in the range 230 nm to 360 nm (2.4.7), a OH O
0.0008 per cent w/v solution shows absorption maxima at about
240 nm, 326 nm and 335 nm; absorbance at about 240 nm, C25H48N6O8,CH4SO3 Mol. Wt. 656.8
about 0.65, at about 326 nm, about 0.4 and at about 335 nm, Desferrioxamine Mesylate is 30-amino-3,14,25-trihydroxy-
about 0.35. 3,9,14,20,25-pentaazatriacontane-2,10,13,21,24-pentaone
C. Gives reaction A of chlorides (2.3.1). methanesulphonate.
379
DESFERRIOXAMINE INJECTION IP 2007
Desferrioxamine Mesylate contains not less than 98.0 per cent point potentiometrically using a platinum indicator electrode
and not more than 102.0 per cent of C25H48N6O8,CH4SO3, and a calomel reference electrode (2.4.25). Towards the end of
calculated on the anhydrous basis. the titration, titrate uniformly and at a rate of about 0.2 ml per
Description. A white or almost white powder. minute. Retain the titrated solution (solution A) for
Identification test B.
Identification 1 ml of 0.1 M ferric ammonium sulphate is equivalent to
0.06568 g of C25H48N6O8,CH4SO3.
B and C may be omitted if tests A and D are carried out. Desferrioxamine Mesylate intended for use in the
Compare the spectrum with that obtained with desferrioxamine
mesylate RS or with the reference spectrum of desferrioxamine
requirement.
mesylate.
B. The titrated solution (solution A) obtained in the Assay is
Unit per mg of desferrioxamine.
reddish brown. The colour is extracted by benzyl alcohol but
not by ether. Desferrioxamine Mesylate intended for use in the
C. Dissolve 5 mg in 5 ml of water, add 2 ml of a 0.5 per cent
w/v solution of tribasic sodium phosphate, mix and then add
0.5 ml of a 2.5 per cent w/v solution of sodium 1,2-
naphthoquinone-4- sulphonate; a blackish brown colour is Sterility (2.2.11). Complies with the test for sterility.
produced. Storage. Store protected from light in a refrigerator (2° to 8°).
D. Dissolve 0.1 g in 5 ml of 2 M hydrochloric acid and add Do not freeze. If the substance is sterile, store in sterile, air-
1 ml of barium chloride solution; the solution remains clear. tight, tamper-evident containers sealed so as to exclude micro-
In a porcelain crucible mix 0.1 g with 1 g of anhydrous sodium organisms.
carbonate, heat and ignite over a Bunsen flame. Allow to Labelling. The label states where applicable, that the
cool, dissolve the residue in 10 ml of water by heating if substance is sterile.
necessary and filter; the filtrate gives reaction A of sulphates
(2.3.1).
Tests Desferrioxamine Injection

Appearance of solution. A 10.0 per cent w/v solution is not Desferrioxamine Mesylate Injection; Deferoxamine
more opalescent than opalescence standard OS2 (2.4.1), and Injection
the absorbance of the solution at about 425 nm is not more Desferrrioxamine Mesylate Injection is a sterile material
than 0.10 (2.4.7). consisting of Desferrrioxamine Mesylate with or without
pH (2.4.24). 3.7 to 5.5, determined in a freshly prepared 10.0 per excipients. It is filled in a sealed container.
cent w/v solution. The injection is constituted by dissolving the contents of the
Heavy metals (2.3.13). 1.0 g complies with the limit test for sealed container in the requisite amount of sterile Water for
heavy metals, Method B (10 ppm). Injections, immediately before use.
Chlorides (2.3.12). 0.75 g complies with the limit test for
chlorides (330 ppm).
Sulphates (2.3.17). 0.25 g complies with the limit test for
Sulphated ash (2.3.18). Not more than 0.1 per cent. recommended by the manufacturer.
Water (2.3.43). Not more than 2.0 per cent, determined on Desferrrioxamine Mesylate Injection contains not less than
1.0 g. 90.0 per cent and not more than 120.0 per cent of the stated
amount of desferrioxamine mesylate, C25H48N6O8,CH4SO3.
water and add 4 ml of 0.05 M sulphuric acid. Titrate slowly Description. A white or almost white powder; very
with 0.1 M ferric ammonium sulphate, determining the end- hygroscopic.
380
IP 2007 DESLANOSIDE
The contents of the sealed container comply with the Deslanoside is 3-[(O-β-D-glucopyranosyl-(1→ 4)-O-2,6-
requirements stated under Parenteral Preparations dideoxy-β -D-ribo-hexopyranosyl-(1→ 4)-O-2,6-dideoxy-β-
(Powders for Injection) and with the following requirements. D-ribo-hexopyranosyl-(1→ 4)-O-2,6-dideoxy-β-D-ribo-
hexopyranosyl)oxy]-12,14-dihydroxy-3β,5β,12β-card-20 (22)-
Identification enolide.
A. Determine by infrared absorption spectrophotometry (2.4.6). Deslanoside contains not less than 95.0 per cent and not more
Compare the spectrum with that obtained with desferrioxamine than 105.0 per cent of C47H74O19, calculated on the dried basis.
mesylate RS or with the reference spectrum of desferrioxamine
Description. White crystals or a fine, crystalline powder;
mesylate.
hygroscopic. It loses water in an atmosphere of low relative
B. The titrated solution (solution A) obtained in the Assay is humidity.
reddish brown. The colour is extracted by benzyl alcohol but
not by ether. Identification
Unit per mg of desferrioxamine. A. Prepare a dispersion of the substance under examination
by dissolving 1 mg in 0.3 ml of methanol and triturating with
Sterility (2.2.11). Comply with the test for sterility. 0.4 g of dry, finely powdered potassium bromide IR until a
Assay. Determine the weight of the contents of 10 containers. uniform and dry mixture is obtained. The powder complies
Weigh accurately a quantity of the mixed contents of the with the following test.
10 containers containing about 0.5 g of Desferrioxamine Determine by infrared absorption spectrophotometry (2.4.6).
Mesylate, dissolve in 25 ml of water and add 4 ml of 0.05 M Compare the spectrum with that obtained with deslanoside
sulphuric acid. Titrate slowly with 0.1 M ferric ammonium RS treated in the same manner. When comparing the spectra
sulphate, determining the end-point potentiometrically using attention should be given to the absence of a distinct maximum
a platinum indicator electrode and a calomel reference electrode at 1260 cm-1 and to the intensity of the maximum at 1740 cm-1.
(2.4.25). Towards the end of the titration, titrate uniformly and
at a rate of about 0.2 ml per minute. Retain the titrated solution B. In the test for Related substances, the principal band in the
(solution A) for Identification test B. chromatogram obtained with test solution (b) corresponds to
1 ml of 0.1 M ferric ammonium sulphate is equivalent to
0.06568 g of C25H48N6O8,CH4SO3. C. Suspend 0.5 mg in 0.5 ml of ethanol (60 per cent) and add
0.1 ml of dinitrobenzoic acid solution and 0.1 ml of 2 M sodium
Storage. Store protected from light in a refrigerator (2° to 8°).
hydroxide; the suspension becomes violet.
Do not freeze.
D. Dissolve 5 mg in 5 ml of glacial acetic acid, add 0.1 ml of
ferric chloride test solution, mix and cautiously add 2 ml of
Deslanoside sulphuric acid so as to form a separate layer; a brown ring is
formed at the junction of the liquids and the upper layer
O O develops a green colour which becomes blue on standing.
HO Tests
H3C
Appearance of solution. A 2.0 per cent w/v solution in a mixture
H3C H of equal volumes of chloroform and methanol is clear (2.4.1),
H OH
Specific optical rotation (2.4.22). +7.0° to +8.5°, determined in
OH O
H a 2.0 per cent w/v solution in dehydrated pyridine.
O
HOH2C O 3 (2.4.17), coating the plate with silica gel G.
OH
OH Mobile phase. A mixture of 130 volumes of dichloromethane,
36 volumes of methanol and 3 volumes of water.
OH
OH Solvent mixture. A mixture of equal volumes of chloroform
C47H74O19 Mol. Wt. 943.1 and methanol.
381
DESLANOSIDE INJECTION IP 2007
Test solution (a). Dissolve 0.2 g of the substance under Identification

examination in 10 ml with solvent mixture.
Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with plate with silica gel G.
36 volumes of methanol and 3 volumes of water.
deslanoside RS in the same solvent mixture.
Reference solution (b). Dilute 5 ml of reference solution (a) to Test solution. Transfer a volume of the injection containing
20 ml with the same solvent mixture. about 2 mg of Deslanoside to a separator and extract with
25 ml of a mixture of 7 volumes of chloroform and 3 volumes of
Reference solution (c). Dilute 5 ml of reference solution (a) to ethanol (95 per cent). Transfer the extract to a 10-ml flask and
50 ml with the same solvent mixture. evaporate to dryness on a water-bath. Dissolve the residue in
Apply separately to the plate, as 1-cm bands, 5 µl of each 1 ml of a mixture of equal volumes of chloroform and methanol.
solution. After development, dry the plate in a current of Reference solution. A 0.2 per cent w/v solution of deslanoside
warm air, spray with ethanolic sulphuric acid (5 per cent RS in the same solvent mixture.
v/v), heat at 140° for 15 minutes and examine in daylight. Any
secondary band in the chromatogram obtained with test Apply separately to the plate, as 1-cm bands, 5 µl of each
solution (a) is not more intense than the band in the solution. After development, dry the plate in a current of
chromatogram obtained with reference solution (b) and not warm air, spray with ethanolic sulphuric acid (5 per cent
more than two such bands are more intense than the band in v/v), heat at 140° for 15 minutes and examine in daylight. The
the chromatogram obtained with reference solution (c). principal spot in the chromatogram obtained with the test
on the residue obtained in the test for Loss on drying.
Loss on drying (2.4.19). Not more than 5.0 per cent, determined Tests
on 0.5 g by drying in an oven at 105° at a pressure of 1.5 to
2.5 kPa. pH (2.4.24). 5.5 to 7.0.
Assay. Protect the solutions from light throughout the assay Other tests. Complies with the tests stated under Parenteral
and maintain at a constant temperature of 20° ± 1°. Preparations (Injections).
Weigh accurately about 30 mg and dissolve in sufficient Assay. Protect the solution from light throughout the assay
methanol to produce 100.0 ml. Dilute 10.0 ml of this solution and maintain at a constant temperature of 20° ± 1°.
to 100.0 ml with methanol. To 10.0 ml of the resulting solution
To an accurately measured volume of the injection containing
add 6 ml of alkaline picric acid solution and dilute to 25.0 ml
3 mg of Deslanoside, add 10 ml of water and extract with five
with water. Allow to stand for 1 hour and measure the
quantities, each of 20 ml, of a mixture of 60 volumes of
chloroform and 40 volumes of 2-propanol with the addition
490 nm (2.4.7), using as the blank a mixture of 10 ml of methanol
of sodium chloride if necessary to disperse any emulsions
and 6 ml of alkaline picric acid solution diluted to 25.0 ml
that may form. Wash each extract with the same quantities of
with water. Calculate the content of C47H74O19 from the
20 ml and then of 10 ml of water. Filter the combined extracts
absorbance obtained by simultaneously carrying out the
through a plug of cotton wool and evaporate the filtrate to
operation using 30 mg of undried deslanoside RS instead of
dryness at about 35° at a pressure not exceeding 0.7 kPa.
the substance under examination.
Transfer the residue to a flask with methanol and add sufficient
Storage. Store protected from light in a refrigerator (2° to 8°). methanol to produce 20.0 ml. To 10.0 ml of the resulting
Do not freeze. solution add 6 ml of alkaline picric acid solution and dilute
to 25.0 ml with water. Allow to stand for 1 hour and measure
Deslanoside Injection about 490 nm (2.4.7), using as blank a mixture of 10.0 ml of
methanol and 6 ml of alkaline picric acid solution diluted to
Deslanoside Injection is a sterile solution of Deslanoside in 25.0 ml with water. Calculate the content of C47H74O19. from
Water for Injections containing suitable buffering agents. the absorbance obtained by simultaneously carrying out the
Deslanoside Injection contains not less than 90.0 per cent operation using a solution prepared by dissolving 30 mg of
and not more than 110.0 per cent of the stated amount of deslanoside RS in sufficient methanol to produce 50.0 ml,
deslanoside, C47H74O19. diluting 25.0 ml to 100.0 ml with methanol and continuing as
382
IP 2007 DESOXYCORTONE ACETATE
described above beginning at the words “To 10.0 ml of the Reference solution (b). Mix equal volumes of the test solution
resulting solution...”. and reference solution (a).
Storage. Store protected from light. Place the dry plate in a tank containing a shallow layer of the
solvent mixture, allow the liquid to ascend to the top, remove
the plate from the tank and allow the solvents to evaporate.
Use within 2 hours, with the flow of the mobile phase in the
Desoxycortone Acetate direction in which the aforementioned treatment was done.
Desoxycorticosterone Acetate; Deoxycortone Acetate Apply to the plate 2 µl of each solution. Allow the mobile
O
O Heat at 120° for a further 10 minutes, allow to cool and examine
O CH3
H3C in daylight and in ultraviolet light at 365 nm. The principal
H3C H
H H reference solution (a). The principal spot in the chromatogram
O
compact spot.
C23H32O4 Mol. Wt. 372.5 D. Dissolve 40 mg in 1 ml of methanol, warm and add 1 ml of
alkaline cupritartrate solution; a red precipitate is formed.
Desoxycortone Acetate is 3,20-dioxo-4-pregnen-21-yl acetate.
E. Dissolve 5 mg in 0.5 ml of methanol, add 0.5 ml of
Desoxycortone Acetate contains not less than 96.0 per cent
ammoniacal silver nitrate solution; a black precipitate is
and not more than 104.0 per cent of C23H32O4, calculated on
slowly produced in the cold but is rapidly produced on
the dried basis.
warming.
Description. A white or creamy-white, crystalline powder;
odourless. Tests
Identification Specific optical rotation (2.4.22). +171.0° to +179.0°, determined

Tests B, D and E may be omitted if tests A and C are carried
(2.4.14).
out.
A. Determine by infrared absorption spectrophotometry (2.4.6). examination in the mobile phase and dilute to 10 ml with the
Compare the spectrum with that obtained with desoxycortone mobile phase.
acetate RS.
Reference solution (a). Dissolve 2 mg of desoxycortone
B. When examined in the range 230 nm to 360 nm (2.4.7), a acetate RS and 2 mg of betamethasone 17-valerate RS in the
0.001 per cent w/v solution in ethanol (95 per cent) shows an mobile phase and dilute to 200 ml with the mobile phase.
absorption maximum at about 240 nm; absorbance at about
240 nm, 0.43 to 0.46. Reference solution (b). Dilute 1 ml of the test solution to
the plate with silica gel G. Chromatographic system
Solvent mixture. A mixture of 90 volumes of acetone and octadecylsilyl silica gel (5 µm),
10 volumes of 1,2-propanediol. – mobile phase: a mixture of 350 ml of water and 600 ml of
Mobile phase. A mixture of equal volumes of cyclohexane acetonitrile, allowed to equilibrate, diluted to 1000 ml
and light petroleum (40° to 60°). with water and mixed again,
Test solution. Dissolve 25 mg of the substance under – spectrophotometer set at 254 nm,
examination in 10 ml of the same solvent mixture. – a 20 µl loop injector.
Reference solution (a). Dissolve 25 mg of desoxycortone Equilibrate the column with the mobile phase for about
acetate RS in 10 ml of the same solvent mixture. 30 minutes.
383
DESOXYCORTONE ACETATE INJECTION IP 2007
Adjust the sensitivity of the system so that the height of the Test solution. Dilute the injection with carbon tetrachloride
principal peak in the chromatogram obtained with reference to give a solution containing 0.25 per cent w/v of
solution (b) is not less than 50 per cent of the full scale of the Desoxycortone Acetate.
recorder. Reference solution. A 0.25 per cent w/v solution of
Inject reference solution (a). The retention times are: desoxycortone acetate RS in carbon tetrachloride.
betamethasone 17-valerate, about 7.5 minutes and
desoxycortone acetate about 9.5 minutes. The test is not valid
dry the plate in air until the odour of solvent is no longer
unless the resolution between the peaks corresponding to
detectable, spray with ethanolic sulphuric acid (10 per cent
betamethasone 17-valerate and desoxycortone acetate is at
v/v), heat at 105° for 30 minutes and examine in ultraviolet
least 4.5. If necessary, adjust the concentration of acetonitrile
in the mobile phase.
Inject the test solution and reference solution (b). Continue chromatogram obtained with the reference solution. Ignore
the chromatography for three times the retention time of the any spots due to the vehicle.
principal peak. In the chromatogram obtained with the test
solution, the sum of the areas of all the peaks other than the Tests
principal peak is not greater than the area of the principal peak
in the chromatogram obtained with reference solution (b) Other tests. Complies with the tests stated under Parenteral
(0.5 per cent). Ignore any peak with an area less than 0.1 times Preparations (Injections).
that of the principal peak in the chromatogram obtained with Assay. To an accurately measured volume containing 10 mg
reference solution (b). of Desoxycortone Acetate add sufficient ethanol to produce
Sulphated ash (2.3.18). Not more than 0.1 per cent. 100.0 ml. Dilute 2.0 ml of this solution to 100.0 ml with ethanol
and mix. Measure the absorbance of the resulting solution
Loss on drying (2.4.19). Not more than 0.5 per cent, determined (2.4.7) at the maximum at about 240 nm. Calculate the content
on 1.0 g by drying in an oven at 105°. of C23H32O4 taking 450 as the specific absorbance at 240 nm.
Assay. Weigh accurately about 0.1 g, dissolve in sufficient Storage. Store protected from light.
ethanol to produce 100.0 ml. Dilute 2.0 ml of this solution to
100.0 ml with ethanol and mix. Measure the absorbance of the Labelling. The label states (1) the composition of the solvent;
resulting solution (2.4.7) at the maximum at about 240 nm. (2) that it is meant for intramuscular injection only; (3) that
Calculate the content of C23H32O4 taking 450 as the specific any sediment should be dissolved by warming before use.
Dexamethasone
Desoxycortone Acetate Injection
O
Desoxycorticosterone Acetate Injection; Deoxycortone H3C OH
Acetate Injection HO OH
H3C H CH3
Desoxycortone Acetate Injection is a sterile solution of
Desoxycortone Acetate in Ethyl Oleate or other suitable ester, F H
in a suitable fixed oil, or in any mixture of these. It may contain
O
suitable alcohols.
Desoxycortone Acetate Injection contains not less than C22H29FO5 Mol. Wt. 392.5
amount of desoxycortone acetate, C23H32O4. Dexamethasone is 9α-fluoro-11β,17α,21-trihydroxy-16α-
methyl-1,4-pregnadiene-3,20-dione.
Identification Dexamethasone contains not less than 96.0 per cent and not
Determine by thin-layer chromatography (2.4.17), coating the more than 104.0 per cent of C22H29FO5, calculated on the dried
plate with silica gel G. basis.
Mobile phase. A mixture of 70 volumes of n-heptane and Description. White or almost white crystals or a crystalline
30 volumes of acetone. powder; odourless.
384
IP 2007 DEXAMETHASONE
Identification Light absorption (2.4.7). Absorbance of a 0.001 per cent w/v

solution in ethanol (95 per cent) at the maximum at about
Test A may be omitted if tests B, C and D are carried out. Tests 240 nm, 0.38 to 0.41.
C and D may be omitted if tests A and B are carried out.
Compare the spectrum with that obtained with dexamethasone
Test solution. To 25 mg of the substance under examination
RS or with the reference spectrum of dexamethasone.
add 1.5 ml of acetonitrile and 5 ml of mobile phase A. Mix with
B. Determine by thin-layer chromatography (2.4.17), coating the aid of ultrasonic sound until the solids are completely
the plate with a suitable silica gel containing a fluorescent dissolved and add sufficient of the mobile phase to produce
indicator with an optimal intensity at about 254 nm. 10 ml and mix well.
Mobile phase. A mixture of 85 volumes of ether, 10 volumes of Reference solution (a). Dissolve 2 mg of dexamethasone RS
toluene and 5 volumes of 1-butanol saturated with water. and 2 mg of methylprednisolone RS in sufficient of the mobile
phase to produce 10 ml.
Solvent mixture. A mixture of 9 volumes of chloroform and 1
volume of methanol. Reference solution (b). Dilute 1 ml of the test solution to
100 ml with mobile phase A
examination in 10 ml with solvent mixture. Chromatographic system
Reference solution (a). A 0.25 per cent w/v solution of octadecylsilyl silica gel (5 µm),
dexamethasone RS in the same solvent mixture. – column temperature. 45°,
Reference solution (b). A solution containing 0.125 per cent – mobile phase: A. 250 volumes of acetonitrile and
w/v each of the substance under examination and 700 volumes of water mixed, allowed to equilibrate and
dexamethasone RS in the same solvent mixture. adjusted to 1000 volumes with water and mixed,
B. acetonitrile,
w/v each of dexamethasone RS and betamethasone RS in the
same solvent mixture.
below,
Apply to the plate 2 µl of each solution. After development, – spectrophotometer set at 254 nm,
dry the plate in air, spray with ethanolic sulphuric acid – a 20 µl loop injector.
(20 per cent v/v), heat at 120° for 10 minutes or until spots
appear, allow to cool and examine in daylight and in ultraviolet
phase A phase B
obtained with the test solution is similar in colour in day-light,
fluorescence in ultraviolet light at 365 nm, in position and size 0 100 0 isocratic
to that in the chromatogram obtained with reference solution 15 100→0 0→100 begin
(a) and the chromatogram obtained with reference solution linear gradient
(b) shows only one spot. The test is not valid unless the
40 0 100 end chromatogram,
chromatogram obtained with reference solution (c) shows two
return to 100 A
spots that are close to one another but separated.
41 100 0 being equilibration
C. Place 2 ml of a 0.01 per cent w/v solution in ethanol in a with A
stoppered tube, add 10 ml of phenylhydrazine-sulphuric acid
46=0 100 0 end equilibration,
solution, mix, place in a water-bath at 60° for 20 minutes and
being next
cool immediately. Absorbance of the resulting solution at the
chromatogram
maximum at about 419 nm, not less than 0.4 (2.4.7).
Equilibrate the column for at least 30 minutes with mobile
D. To 2 ml of sulphuric acid add 2 mg and shake to dissolve;
phase B and then with mobile phase A for 5 minutes. For
a faint reddish brown colour is produced within 5 minutes.
subsequent operations use the conditions described from
Add 10 ml of water and mix; the colour is discharged.
40 to 46 minutes.
Tests Adjust the sensitivity of the system so that the height of the
Specific optical rotation (2.4.22). +75.0° to +80.0°, determined solution (b) is at least 50 per cent of the full scale of the
in a 1.0 per cent w/v solution in dioxan. recorder.
385
DEXAMETHASONE TABLETS IP 2007
Inject reference solution (a). When the chromatograms are Mobile phase. A mixture of 85 volumes of ether, 10 volumes of
recorded, the retention times are; methylprednisolone about toluene and 5 volumes of 1-butanol saturated with water.
11.5 minutes, and dexamethasone about 13 minutes. The test Test solution. Dissolve 25 mg of the substance under
is not valid unless the resolution between the peaks examination in 10 ml of a mixture of 9 volumes of chloroform
corresponding to methylprednisolone and dexamethasone is and 1 volume of methanol.
at least 2.8; if necessary, adjust the concentration of acetonitrile
in mobile phase A. Reference solution (a). A 0.25 per cent w/v solution of
dexamethasone RS in the same solvent mixture.
Inject mobile phase A as the blank, the test solution and
reference solution (b). Record the chromatogram of the test Reference solution (b). A solution containing 0.125 per cent
solution for twice the retention time of the principal peak. In w/v each of the substance under examination and
the chromatogram obtained with the test solution, the area of dexamethasone RS in the same solvent mixture.
any peak other than the principal peak, is not greater than Reference solution (c). A solution containing 0.125 per cent
0.5 times the area of the principal peak in the chromatogram w/v each of dexamethasone RS and betamethasone RS in the
obtained with reference solution (b) (0.5 per cent); the sum of same solvent mixture.
the areas of all the peaks other than the principal peak, is not Apply to the plate 2 µl of each solution. After development,
greater than the area of the principal peak in the chromatogram dry the plate in air, spray with ethanolic sulphuric acid
obtained with reference solution (b) (1 per cent). Ignore any (20 per cent v/v), heat at 120° for 10 minutes or until spots
peak due to the blank and any peak with an area less than appear, allow to cool and examine in daylight and in ultraviolet
0.05 times the area of the principal peak in the chromatogram light at 365 nm. The principal spot in the chromatogram
obtained with reference solution (b). obtained with the test solution is similar in colour in day-light,
Sulphated ash (2.3.18). Not more than 0.1 per cent. fluorescence in ultraviolet light at 365 nm, in position and size
to that in the chromatogram obtained with reference solution
(a) and the chromatogram obtained with reference solution
(b) shows only one spot. The test is not valid unless the
chromatogram obtained with reference solution (c) shows two
Assay. Weigh accurately about 0.1 g and dissolve in sufficient spots that are close to one another but separated.
ethanol to produce 100.0 ml and mix. Dilute 2.0 ml of this
C. To 2 ml of sulphuric acid add 2 mg and shake to dissolve;
solution to 100.0 ml with ethanol and mix well. Determine the
a faint reddish brown colour is produced within 5 minutes.
absorbance of the resulting solution (2.4.7) at the maximum at
Add 10 ml of water and mix; the colour is discharged.
about 238 nm. Calculate the content of C22H29FO5 taking 394
as the specific absorbance at 238 nm. Tests
Storage. Store protected from light. Related substances. Determine by liquid chromatography
(2.4.14).
Test solution. To a weighed quantity of the tablets containing
Dexamethasone Tablets 2.5 mg of Dexamethasone add 10 ml of acetonitrile, mix with
the aid of ultrasound and filter through a 0.45 µm filter. Dilute
Dexamethasone Tablets contain not less than 90.0 per cent
4 ml of the filtrate to 10 ml with water.
dexamethasone, C22H29FO5. Reference solution (a). Dilute 1 ml of the test solution to
Identification Reference solution (b). Dissolve 2 mg of dexamethasone RS
Shake a quantity of the powdered tablets containing 20 mg of and 2 mg of methylprednisolone RS in mobile phase A and
Dexamethasone with 50 ml of chloroform for 30 minutes, filter dilute to 100 ml with the same solvent.
and evaporate the filtrate to dryness at 105° for 2 hours. The Chromatographic system
residue complies with the following tests. – a stainless steel column 25 cm x 4.6 mm, packed with
A. Determine by infrared absorption spectrophotometry (2.4.6). octadecylsilyl silica gel (5 µm) (such as Hypersil ODS),
Compare the spectrum with that obtained with dexamethasone – column temperature. 45°,
RS or with the reference spectrum of dexamethasone. – mobile phase: A. 15 per cent v/v acetonitrile,
B. acetonitrile,
B. Determine by thin-layer chromatography (2.4.17), coating – flow rate. 2.5 ml per minute,
the plate with a suitable silica gel containing a fluorescent – a linear gradient programme using the conditions given
indicator with an optimal intensity at about 254 nm. below,
386
IP 2007 DEXAMETHASONE SODIUM PHOSPHATE
– spectrophotometer set at 254 nm, – flow rate. 1.4 ml per minute,

– a 20 µl loop injector. – spectrophotometer set at 238 nm,
phase A phase B Calculate the content of C22H29FO5 in the tablet.
(min) (per cent v/v) (per cent v/v) Other tests. Comply with the tests stated under Tablets.
0 100 0 isocratic
Assay — For tablets containing 2 mg or more of
15 100→0 0→100 begin dexamethasone
linear gradient
Determine by liquid chromatography (2.4.14), protected from
40 0 100 end chromatogram, light.
return to 100 A
Test solution (a). Weigh and powder 20 tablets. To a quantity
41 100 0 being equilibration of the powder containing about 2.5 mg of Dexamethasone add
with A 20.0 ml of methanol (50 per cent), shake for 20 minutes and
46=0 100 0 end equilibration, filter through a glass-fibre filter paper (such as Whatman
being next GF/C).
chromatogram
Test solution (b). Prepare in the same manner as test solution
Inject reference solution (b). When the chromatograms are (a) but use 20.0 ml of a 0.01 per cent w/v solution of
recorded, the retention times are; methylprednisolone about hydrocortisone in methanol (50 per cent) in place of the
13 minutes, and dexamethasone about 16 minutes. The test is 20.0 ml of methanol (50 per cent).
not valid unless the resolution between the peaks
corresponding to methylprednisolone and dexamethasone is
of dexamethasone RS and 0.01 per cent w/v of hydrocortisone
at least 2.8; if necessary, adjust the concentration of acetonitrile
(internal standard) in methanol (50 per cent).
in mobile phase A.
Inject mobile phase A, the test solution and reference solution
(a). In the chromatogram obtained with the test solution, the
octadecylsilyl silica gel (5 µm) (such as (Spherisorb
area of any secondary peak is not greater than 0.5 times the
ODS 1),
– mobile phase: a mixture of 53 volumes of water and
reference solution (a) (0.5 per cent); the sum of the areas of all
secondary peaks is not greater than the area of the principal
(a) (1.0 per cent). Ignore any peak due to mobile phase A and
any peak with an area less than 0.05 times the area of the
principal peak in the chromatogram obtained with reference Calculate the content of C22H29FO5 in the tablets.
solution (a) (0.05 per cent). For tablets containing less than 2 mg of dexamethasone.
Uniformity of content. Comply with the test stated under Use the average of the ten individual results obtained in the
Tablets. test for Uniformity of content.
Test solution. Finely crush one tablet, add sufficient quantity Storage. Store protected from light.
of a 0.002 per cent w/v solution of hydrocortisone in methanol
(50 per cent) to produce a solution containing 0.0025 per cent
w/v solution of Dexamethasone, shake for 10 minutes and
filter through a glass-fibre filter paper (such as Whatman
Dexamethasone Sodium Phosphate
GF/C).
ONa
Reference solution. A solution containing 0.0025 per cent w/v O
H3C O P O
of dexamethasone RS and 0.002 per cent w/v of hydrocortisone HO OH ONa
(internal standard) in methanol (50 per cent).
H3C H CH3
– a stainless steel column 20 cm x 5 mm, packed with octade- F H
cylsilyl silica gel (10 µm) (such as (Spherisorb ODS 1), O
– mobile phase: a mixture of 53 volumes of water and
47 volumes of methanol, C22H28FNa2O8P Mol. Wt. 516.4
387
DEXAMETHASONE SODIUM PHOSPHATE IP 2007
Dexamethasone Sodium Phosphate is disodium 9α-fluoro- 0.00143 per cent w/v solution of potassium dihydrogen
11β,17α-dihydroxy-16α-methyl-3,20-dioxo-1,4-pregna- dien- phosphate in a similar manner.
21-yl phosphate. Free dexamethasone. Not more than 1 per cent w/w, determined
Dexamethasone Sodium Phosphate contains not less than by the following method. Weigh accurately about 25 mg in a
97.0 per cent and not more than 103.0 per cent of C22H28FNa2O8P, glass-stoppered 50-ml tube, add 5 ml of water and shake to
calculated on the anhydrous, and ethanol-free or solvent-free dissolve. Add 25.0 ml of dichloromethane, insert the stopper
basis. and mix by gentle shaking. Allow to stand until the
dichloromethane layer is clear. Measure the absorbance of
Description. A white or slightly yellow, crystalline powder;
the dichloromethane solution at the maximum at about 236 nm
almost odourless; very hygroscopic. It shows polymorphism.
(2.4.7), using dichloromethane as the blank. Calculate the
Identification content of dexamethasone taking 390 as the specific
absorbance at about 236 nm.
Test A may be omitted if tests B, C and, D are carried out. Related substances. Determine by liquid chromatography
Tests B and D may be omitted if Tests A and C are carried out. (2.4.14).
A. Determine by infrared absorption spectrophotometry (2.4.6). Test solution. Dissolve 25 mg of the substance under
Compare the spectrum with that obtained with dexamethasone examination in the mobile phase and dilute to 10 ml with the
sodium phosphate RS or with the reference spectrum of mobile phase.
dexamethasone sodium phosphate.
Reference solution (a). Dissolve 2 mg of dexamethasone
B. Dissolve 10 mg in 5 ml of water and dilute to 100 ml with sodium phosphate RS and 2 mg of betamethasone sodium
ethanol. To 2 ml of the resulting solution in a glass-stoppered phosphate RS in the mobile phase and dilute to 100 ml with
tube add 10 ml of phenylhydrazine- sulphuric acid solution, the mobile phase.
mix, heat in a water-bath at 60° for 20 minutes and cool
immediately. Absorbance of the resulting solution at the Reference solution (b). Dilute 1 ml of the test solution to
maximum at about 419 nm, not less than 0.20 (2.4.7). 100 ml with the mobile phase.
C. In the test for Related substances, the principal peak in the Reference solution (c). A 0.0025 per cent w/v solution of
chromatogram obtained with reference solution (b) dexamethasone RS in the mobile phase.
corresponds to the peak in the chromatogram obtained with Chromatographic system
reference solution (c). – a stainless steel column 25 cm x 4.6 mm, packed with
D. Heat gently 40 mg with 2 ml of sulphuric acid until white octadecylsilyl silica gel (5 µm),
fumes are evolved, add nitric acid dropwise until oxidation is – mobile phase: a mixture of 1.360 g of potassium
complete and cool. Add 2 ml of water, heat until white fumes dihydrogen phosphate and 0.60 g of hexylamine allowed
are evolved again, cool, add 10 ml of water and neutralise to to stand for 10 minutes and then dissolved in 182.5 ml of
litmus paper with 5 M ammonia. The solution gives reaction water and 67.5 ml of acetonitrile, mixed and filtered,
A of sodium salts and reaction B of phosphates (2.3.1). – flow rate. 1 ml per minute,
Tests
Equilibrate the column with the mobile phase for about
pH (2.4.24). 7.5 to 9.5, determined in a 1.0 per cent w/v solution. 45 minutes.
Specific optical rotation (2.4.22). +75.0° to +83.0°, determined Adjust the sensitivity of the system so that the height of the
in a 1.0 per cent w/v solution. principal peak in the chromatogram obtained with reference
Inorganic phosphates. Not more than 0.5 per cent, calculated solution (b) is at least 50 per cent of the full scale of the
as PO4, determined by the following method. Weigh accurately recorder.
about 25 mg, dissolve in 10 ml of water, add 4 ml of dilute Inject reference solution (a). The retention times are:
sulphuric acid, 1 ml of ammonium molybdate solution and betamethasone sodium phosphate, about 12.5 minutes and
2 ml of methylaminophenol with sulphite solution and allow dexamethasone sodium phosphate about 14 minutes. The test
to stand for 15 minutes. Add sufficient water to produce is not valid unless the resolution between the peaks
25.0 ml, allow to stand for further 15 minutes and measure the corresponding to betamethasone sodium phosphate and
absorbance of the resulting solution at the maximum at about dexamethasone sodium phosphate is at least 2.2. If necessary,
730 nm (2.4.7). Calculate the content of phosphate from a adjust the concentration of acetonitrile or increase the
calibration curve prepared by treating suitable aliquots of a concentration of water in the mobile phase.
388
IP 2007 DEXAMETHASONE INJECTION
Inject the test solution and reference solution (b). Continue Dexamethasone Injection
peak. In the chromatogram obtained with the test solution: Dexamethasone Sodium Phosphate Injection
the area of any peak other than the principal peak, is not Dexamethasone Injection is a sterile solution of
greater than half the area of the principal peak in the Dexamethasone Sodium Phosphate in Water for Injections.
cent); the sum of the areas of all such peaks is not greater than Dexamethasone Injection contains not less than 95.0 per cent
the area of the principal peak in the chromatogram obtained and not more than 105.0 per cent of the stated amount of
with reference solution (b) (1 per cent). Ignore any peak with dexamethasone phosphate, C22H30FO8P.
an area less than 0.05 times that of the principal peak in the
Identification
Ethanol. Not more than 3.0 per cent w/w, determined by gas
plate with silica gel G.
chromatography (2.4.13).
Internal standard. A 1.0 per cent v/v solution of 1-propanol.
50 volumes of acetone and 1 volume of water.
Test solution (a). A 10.0 per cent w/v solution of the substance Test solution. Dilute a quantity of the injection containing
under examination in water. about 5 mg of dexamethasone phosphate with 25 ml of water
Test solution (b). A solution containing 10.0 per cent w/v of and extract with two quantities, each of 25 ml, of
the substance under examination and 1.0 per cent v/v of the dichloromethane. Discard the dichloromethane each time and
internal standard. transfer the aqueous layer to a 50-ml volumetric flask, dilute to
volume with water and mix. Pipette 5 ml into a 50-ml glass-
Reference solution. A solution containing 1.0 per cent v/v of
stoppered tube and incubate at 37° for 45 minutes with 5 ml of
1-propanol and 1.0 per cent v/v of ethanol. Adjust the content
alkaline phosphatase solution. Extract with 25 ml of
of ethanol to produce a peak of similar height to the
dichloromethane, evaporate 15 ml of the dichloromethane
corresponding peak in the chromatogram obtained with test
extract to dryness and dissolve the residue in 1 ml of
solution (a).
dichloromethane.
Reference solution. Dissolve 3 mg of dexamethasone RS in
– a glass column 1 m x 3.2 mm, packed with porous polymer
sufficient dichloromethane to produce 10 ml.
beads (150 to 180 µm),
– temperature: Apply to the plate 5 µl of each solution. After development,
column. 150°, dry the plate in air until the odour of solvent is no longer
inlet port. 250°, detectable, spray with a 50 per cent w/v solution of sulphuric
detector. 280°, acid, heat at 105° until brown and black spots appear. The
– flow rate. 30 ml per minute of the carrier gas. principal spot in the chromatogram obtained with the test
Inject 2 µl of each solution. with the reference solution.
Calculate the percentage w/w of ethanol assuming the weight
per ml at 25° to be 0.787 g. Tests
Water (2.3.43). Determine on 0.2 g. pH (2.4.24). 7.5 to 8.5.
Total ethanol and water. Not more than 16.0 per cent w/w, Free dexamethasone. Determine by liquid chromatography
calculated from the sum of the percentage of ethanol (2.4.14).
determined by the method described above and the percentage Test solution. Dilute a volume of the injection with the mobile
of water. phase to produce a solution containing the equivalent of
0.25 per cent w/v of dexamethasone phosphate.
water to produce 100.0 ml. Dilute 5.0 ml of this solution to Reference solution (a). A 0.00125 per cent w/v solution of
250.0 ml with water and measure the absorbance of the resulting dexamethasone RS in the mobile phase.
solution at the maximum at about 240 nm (2.4.7). Calculate the Reference solution (b). A solution containing 0.25 per cent
content of C 22H 28FNa 2O 8P, taking 297 as the specific w/v of dexamethasone phosphate RS, 0.01 per cent w/v of
absorbance at 240 nm. propyl hydroxybenzoate and 0.001 per cent w/v of
Storage. Store protected from light. dexamethasone RS in the mobile phase.
389
DEXTRAN 40 INJECTION IP 2007
Reference solution (c). A solution containing 0.01 per cent – spectrophotometer set at 254 nm,
w/v of propyl hydroxybenzoate in the mobile phase. – a 20 µl loop injector.
Chromatographic system Inject reference solution (b). The test is not valid unless the
– a stainless steel column 25 cm x 4.6 mm, packed with resolution between the peaks corresponding to betamethasone
octadecylsilyl silica gel (5 µm) (such as Hypersil ODS), sodium phosphate and dexamethasone phosphate is at least
– mobile phase: a mixture of 1.360 g of potassium 2.2. If necessary, adjust the concentration of acetonitrile or
dihydrogen phosphate and 0.60 g of hexylamine allowed increase the concentration of water in the mobile phase.
to stand for 10 minutes and then dissolved in 182.5 ml of
water and 67.5 ml of acetonitrile, mixed and filtered,
– flow rate. 1 ml per minute, Calculate the content of C22H30FO8P in the injection.
– spectrophotometer set at 254 nm, Storage. Store protected from light at a temperature not
– a 20 µl loop injector. exceeding 30°.
Equilibrate the column with the mobile phase for about Labelling. The label states the strength in terms of the
45 minutes. equivalent amount of dexamethasone phosphate in a suitable
Adjust the sensitivity of the system so that the height of the dose-volume.
solution (b) is at least 50 per cent of the full scale of the
recorder. Dextran 40 Injection
peak corresponding to dexamethasone is completely separated
Dextran 40 Intravenous Infusion
from the peaks due to dexamethasone phosphate and propyl Dextran 40 Injection is a sterile solution, in Dextrose Injection
hydroxybenzoate. If necessary, adjust the concentration of or in Sodium Chloride Injection, of dextrans of average
acetonitrile or increase the concentration of water in the mobile molecular weight of about 40,000, derived from the dextrans
phase. produced by the fermentation of sucrose by means of a certain
Inject the test solution and reference solution (a). Continue strain of Leuconostoc mesenteroides. The dextrans are
the chromatography for twice the retention time of the principal polymers of dextrose in which the linkages between the
peak. In the chromatogram obtained with the test solution the dextrose units are almost entirely of the α-1→6 type.
area of the peak corresponding to dexamethasone is not greater Dextran 40 Injection contains not less than 9.0 per cent and
than that of the principal peak in the chromatogram obtained not more than 11.0 per cent w/v of dextrans.
Description. An almost colourless, slightly viscous solution.
Preparations (Injections). Tests
Assay. Determine by liquid chromatography (2.4.14). pH (2.4.24). 3.5 to 6.5 for solutions in Dextrose Injection; 4.0 to
Test solution. Dilute an accurately measured volume of the 7.0 for solutions in Sodium Chloride Injection.
injection containing about 8 mg of dexamethasone phosphate Molecular size. For solutions in Dextrose Injection, before
to 100.0 ml with the mobile phase and mix. proceeding with tests A, B and C add 4 volumes of ethanol
Reference solution (a). A 0.008 per cent w/v solution of (95 per cent), centrifuge and dissolve the residue in a volume
dexamethasone sodium phosphate RS in the mobile phase. of Sodium Chloride Injection sufficient to restore the original
volume.
w/v each of dexamethasone phosphate RS and betamethasone A (2.4.28). Determine the viscosity ratios by Method A, using
sodium phosphate RS in the mobile phase. size C U-tube viscometer at 37°, of solutions in saline solution
containing about 3.5, 2.5, 1.5 and 0.75 per cent w/v of dextrans,
Chromatographic system accurately determined. For each solution, plot (viscosity ratio
– a stainless steel column 25 cm x 4.6 mm, packed with – 1.00)/concentration (in per cent w/v) against concentration
octadecylsilyl silica gel (5 µm) (such as Hypersil ODS), (in per cent w/v). The intercept on the viscosity axis of the
– mobile phase: a mixture of 1.360 g of potassium straight line joining the points represents the intrinsic
dihydrogen phosphate and 0.6 g of hexylamine allowed viscosity; the intrinsic viscosity is 0.16 to 0.20.
to stand for 10 minutes and then dissolved in 182.5 ml of
water and 67.5 ml of acetonitrile, mixed and filtered, B. Place in each of five stoppered flasks 100 ml of a solution in
– flow rate. 1 ml per minute, saline solution containing 6 per cent w/v of dextrans and add
390
IP 2007 DEXTRAN 40 INJECTION
slowly, with continuous stirring, sufficient ethanol to produce and 5 ml of strong ammonia solution, and allow to stand for
a faint cloudiness (about 45 ml is usually required). Add 0.5, 10 minutes. Any purple colour produced is not more intense
1.0, 1.5, 2.0 and 2.5 ml of ethanol to the separate flasks, stopper than that produced by treating in the same manner 10 ml of a
the flasks and immerse in a water-bath at about 35° with 0.02 per cent v/v solution of acetone.
occasional shaking until clear solutions are obtained. Transfer
Ethanol. Distil 100 ml, collect the first 45 ml of distillate and
the flasks to a water-bath maintained at 25.0° ± 0.1° and allow
dilute to 50 ml with water. Mix 10 ml of 0.0167 M potassium
to stand overnight or until two clear liquid phases are formed.
dichromate and 10 ml of sulphuric acid in a stoppered boiling
Reject the supernatant liquids, dissolve separately the syrupy
tube, immediately add 5 ml of the distillate, mix, stopper the
residues in sufficient saline solution to produce 25.0 ml,
tube, and allow to stand for 5 minutes. Transfer to a 500-ml
remove the ethanol by evaporation at a pressure of about
flask, dilute to about 300 ml with carbon dioxide-free water,
2 kPa, dilute to 25.0 ml with water and determine the optical
add 2 g of potassium iodide and 1 ml of a 10 per cent w/v
rotation (2.4.22). From the optical rotations calculate the
solution of potassium thiocyanate, allow to stand for
amount of dextrans precipitated as described in the Assay.
5 minutes and titrate the liberated iodine with 0.1 M sodium
Choose that fraction containing as nearly as possible but not
thiosulphate using starch solution, added towards the end
more than 10 per cent of the dextrans present in the injection
of the titration, as indicator. Repeat the determination
and determine its intrinsic viscosity by the method described
beginning at the words “Mix 10 ml of 0.0167 M potassium
under test A; the intrinsic viscosity is not more than 0.27.
dichromate..” but using 5 ml of water in place of 5 ml of the
C. Place in each of four stoppered flasks 100 ml of a solution in distillate. The difference between the titrations is not more
saline solution containing 6 per cent w/v of dextrans and add than 4.2 ml.
slowly, with continuous stirring, 80, 90, 100 and 110 ml Heavy metals (2.3.13). To 4.0 ml add 5 ml of dilute acetic acid
respectively of ethanol. Stopper the flasks, transfer to a water- and sufficient water to produce 25.0 ml. The resulting solution
bath maintained at 25.0° ± 0.1° and allow to stand overnight or complies with the limit test for heavy metals, Method A
until two clear liquid phases are formed. Separate the (5 ppm).
supernatant solution from the syrupy residues. Remove the
ethanol from each supernatant solution separately by Nitrogen (2.3.30). Determine by Method B, using 50 ml. For
evaporation at a pressure of 2 kPa, dialyse in cellophane tubing solutions in Dextrose Injection, use 30 ml of nitrogen-free
against water to remove sodium chloride, adjust the volume sulphuric acid. For solutions in Sodium Chloride Injection
to 25.0 ml with water, add sufficient sodium chloride to use 20 ml of nitrogen-free sulphuric acid.
produce solutions containing 0.9 per cent w/v and determine Not more than 0.35 ml of 0.05 M sulphuric acid is required.
the optical rotation (2.4.22). From the optical rotations,
calculate the amounts of dextrans present as described in the Sulphated ash. Titrate 25 ml with 0.1 M silver nitrate using
Assay. Choose that fraction containing as nearly as possible potassium chromate solution as indicator. Deduct the
but not more than 10 per cent of the dextrans present in the theoretical value of the sulphated ash due to the sodium
injection and determine the intrinsic viscosity by the method chloride present.
in test A above; the intrinsic viscosity is not less than 0.08. 1 ml of 0.1 M silver nitrate is equivalent to 0.007102 g of
Content of dextrose. For solutions in Dextrose Injection, sulphated ash (0.05 per cent w/v)
4.5 to 5.5 per cent w/v, determined by the following method. Foreign protein. Inject 0.5 ml on three occasions at intervals
Dilute 15.0 ml to 50.0 ml with water. To 5.0 ml in a stoppered of 2 days into the peritoneal cavity of each of six healthy
flask add 25 ml of a buffer solution containing 14.3 per cent guinea-pigs weighing not less than 250 g, which have not
w/v of sodium carbonate and 4.0 per cent w/v of potassium previously been treated with any material that will interfere
iodide and 25.0 ml of 0.05 M iodine. Stopper the flask and with the test. Inject 0.2 ml intravenously into each of the three
allow to stand for exactly 30 minutes at 20°, add 30 ml of dilute guinea-pigs 14 days after the first intra-peritoneal injection,
hydrochloric acid and titrate immediately with 0.1 M sodium and into each of the other three guinea-pigs 21 days after the
thiosulphate. Repeat the operation beginning at the words first intra-peritoneal injection. Observe the guinea-pigs for
“add 25 ml of a buffer solution...” but using 5 ml of water in 30 minutes after each intravenous injection and again 24 hours
place of 5 ml of the preparation under examination. The later; the animals exhibit no signs of anaphylaxis such as
difference between the titrations represents the amount of coughing, bristling of hair or respiratory distress.
iodine required to oxidise the dextrose.
1 ml of 0.05 M iodine is equivalent to a 0.00901 g of dextrose. Units per ml.
Acetone. To 10 ml add sufficient ammonium sulphate to give Other tests. Complies with the tests stated under Parenteral
a saturated solution, add 1 ml of sodium nitroprusside solution Preparations (Infusions).
391
Assay. For solutions in Dextrose Injection — Add a drop of a faint cloudiness (about 45 ml). To the separate flasks add
dilute ammonia solution to the required volume and determine 0.5, 1.0, 1.5, 2.0 and 2.5 ml of ethanol, stopper the flasks and
the optical rotation (2.4.22). Calculate the content of dextrans immerse in a water-bath at about 35°, shaking occasionally,
from the following expression 0.5076(α - 0.528D), where α is until clear solutions are obtained. Transfer the flasks to a
the observed angular rotation and D the content of dextrose water-bath maintained at 25.0° ± 0.1° and allow to stand
per cent w/v, determined in the test for Content of dextrose. overnight or until two clear liquid phases are formed. Discard
For solutions in Sodium Chloride Injection — Measure the the supernatant liquids, dissolve separately the syrupy
optical rotation (2.4.22), and multiply the value obtained by residues in sufficient saline solution to produce 25.0 ml, remove
0.5076. the ethanol by evaporation at a pressure of 2 kPa, dilute to
25.0 ml with water and determine the optical rotation (2.4.22).
Storage. Store at a temperature not exceeding 30°. The injection From the optical rotations, calculate the amounts of dextrans
should not be exposed to undue fluctuations of temperature. precipitated as described in the Assay. Choose that fraction
Labelling. The label states (1) the strength as the percentage containing as nearly as possible but not more than 10 per cent
w/v of dextrans; (2) the name of the solvent; (3) the strain of of the dextrans present in the injection and determine its
Leuconostoc mesenteroides used; (4) that the injection should intrinsic viscosity by the method described under test A using
not be used if it is cloudy or if a deposit is present. a U-tube viscometer (size A). The intrinsic viscosity is not
more than 0.36.
C. Place in each of four stoppered flasks 100 ml of a solution in
Dextran 70 Injection saline solution containing 6 per cent w/v of dextrans and add
slowly, with continuous stirring, 80, 90, 100 and 110 ml
Dextran 70 Intravenous Infusion respectively of ethanol. Stopper the flasks, transfer to a water-
Dextran 70 Injection is a sterile solution, in Dextrose Injection bath maintained at 25.0° ± 0.1° and allow to stand overnight or
or in Sodium Chloride Injection, of dextrans of average until two clear liquid phases are formed. Separate the
molecular weight about 70,000, derived from the dextrans supernatant solutions from the syrupy residues. Remove the
produced by the fermentation of sucrose by means of a certain ethanol from each supernatant solution separately by
strain of Leuconostoc mesenteroides. The dextrans are evaporation at a pressure of 2 kPa, dialyse in cellophane tubing
polymers of dextrose in which the linkages between the against water to remove sodium chloride, adjust the volume
dextrose units are almost entirely of the α-1→6 type. to 25.0 ml with water, add sufficient sodium chloride to
produce solutions containing 0.9 per cent w/v of sodium
Dextran 70 Injection contains not less than 5.5 per cent and chloride and determine the optical rotation (2.4.22). From the
not more than 6.5 per cent w/v of dextrans. optical rotations, calculate the amounts of dextrans present
Description. An almost colourless, slightly viscous solution. as described in the Assay. Choose that fraction containing as
nearly as possible but not more than 10 per cent of the dextrans
Tests present in the injection and determine the intrinsic viscosity
pH (2.4.24). 3.5 to 6.5 for solutions in Dextrose Injection; 5.0 to by the method in test A. The intrinsic viscosity is not less
7.0 for solutions in Sodium Chloride Injection. than 0.13.
Molecular size. For solutions in Dextrose Injection, before Content of dextrose (if present). For solutions in Dextrose
proceeding with tests A, B and C, add 4 volumes of ethanol Injection, between 4.5 and 5.5 per cent w/v, determined by the
(95 per cent), centrifuge and dissolve the residue in sufficient following method. Dilute 15.0 ml to 50.0 ml with water. To
Sodium Chloride Injection to restore the original volume. 5.0 ml in a stoppered flask, add 25 ml of a buffer solution
containing 14.3 per cent w/v of sodium carbonate and 4.0 per
A (2.4.28). Determine the viscosity ratios by Method A, using cent w/v of potassium iodide, and 25.0 ml of 0.05 M iodine.
a size C U-tube viscometer at 37°, of solutions in saline solution Stopper the flask, allow to stand for exactly 30 minutes at 20°,
containing about 3.5, 2.5, 1.5 and 0.75 per cent w/v of dextrans, add 35 ml of dilute hydrochloric acid and titrate immediately
accurately determined. For each solution, plot (viscosity ratio with 0.1 M sodium thiosulphate. Repeat the operation using
–1.00)/concentration (in percentage w/v) against concentration 5 ml of water and beginning at the words ‘add 25 ml of a buffer
(in per cent w/v). The intercept on the viscosity ratio axis of a solution…...’. The difference between the titrations represents
straight line through the points represents the intrinsic the amount of iodine required to oxidise the dextrose.
viscosity. The intrinsic viscosity is 0.22 to 0.27.
1 ml of 0.05 M iodine is equivalent to 0.00901 g of C6H12O6.
B. Place 100 ml in each of five stoppered flasks and adjust the
temperature to 25.0 ± 0.1°. Maintaining this temperature, add Acetone. To 10 ml add sufficient ammonium sulphate to give
slowly with continuous stirring sufficient ethanol to produce a saturated solution, add 1 ml of sodium nitroprusside solution
392
IP 2007 DEXTRAN 110 INJECTION
and 5 ml of strong ammonia solution, and allow to stand for guinea-pigs 14 days after the first intra-peritoneal injection,
10 minutes. Any purple colour produced is not more intense and into each of the other three guinea-pigs 21 days after the
than that produced by treating in the same manner 10 ml of a first intraperitoneal injection. Observe the guinea-pigs for
0.02 per cent v/v solution of acetone in the same solvent. 30 minutes after each intravenous injection and again 24 hours
later; the animals exhibit no signs of anaphylaxis such as
Content of sodium chloride (if present). For solutions in Sodium
coughing, bristling of hair or respiratory distress.
Chloride Injection, 0.81 to 0.99 per cent w/v, determined by the
following method. Titrate an accurately measured volume Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit
containing 0.1 g of sodium chloride with 0.1 M silver nitrate per ml.
using potassium chromate solution as indicator. Other tests. Complies with the tests stated under Parenteral
1 ml of 0.1 M silver nitrate is equivalent to 0.005844 g of NaCl. Preparations (Infusions).
5-Hydroxymethylfurfural and related substances (if dextrose Assay. For solutions in Dextrose Injection — Add a drop of
is present). Dilute a volume containing 1.0 g of Dextrose to dilute ammonia solution to 25.0 ml of the injection under
500.0 ml with water and measure the absorbance of the resulting examination and determine the optical rotation (2.4.22).
solution at the maximum at about 284 nm; absorbance at about Calculate the content of dextrans from the expression
284 nm, not more than 0.25 (2.4.7). 0.5076(α – 0.528) D, where α is the observed angular rotation
and D the content of dextrose, in per cent w/v, determined in
Ethanol. Distil 100 ml, collect the first 45 ml of the distillate and
the test for Content of dextrose.
dilute to 50 ml with water. Mix 10 ml of 0.0167 M potassium
dichromate and 10 ml of sulphuric acid in a stoppered boiling For solutions in Sodium Chloride Injection — Measure the
tube, immediately add 5.0 ml of the distillate, mix, stopper the optical rotation (2.4.22), and multiply the value obtained by
tube, and allow to stand for 5 minutes. Transfer to a 500-ml 0.5076.
flask, dilute to about 300 ml with carbon dioxide-free water, Storage. Store in single dose containers in a cool place. The
add 2 g of potassium iodide and 1 ml of 10 per cent w/v injection should not be exposed to undue fluctuations of
solution of potassium thiocyanate, allow to stand for 5 minutes temperature.
and titrate the liberated iodine with 0.1 M sodium thiosulphate
using starch solution, added towards the end of the titration, Labelling. The label states (1) the strength as the percentage
as indicator. Repeat the determination beginning at the words w/v of dextrans; (2) the name of the solvent; (3) the strain of
‘Mix 10 ml of 0.0167 M potassium dichromate…..’ but using Leuconostoc mesenteroides used; (4) that the injection should
5.0 ml of water in place of 5.0 ml of the distillate. The difference not be used if it is cloudy or if a deposit is present.
between the titrations is not more than 4.2 ml.
Heavy metals (2.3.13). Not more than 5 ppm, determined by
Method A, on 4.0 ml to which 5 ml of dilute acetic acid and Dextran 110 Injection
sufficient water are added to produce 25.0 ml.
Dextran 110 Intravenous Infusion
Nitrogen. Carry out Method B for the determination of nitrogen
(2.3.30), using 50.0 ml. For solutions in Dextrose Injection, use Dextran 110 Injection is a sterile solution, in Dextrose Injection
30 ml of nitrogen-free sulphuric acid. For solutions in Sodium or in Sodium Chloride Injection, of dextrans of average
Chloride Injection, use 20 ml of nitrogen-free sulphuric acid. molecular weight of about 110,000, derived from the dextrans
Not more than 0.35 ml of 0.05 M sulphuric acid is required. produced by the fermentation of sucrose by means of a certain
strain of Leuconostoc mesenteroides. The dextrans are
Sulphated ash (2.3.18). Not more than 0.05 per cent w/v, polymers of dextrose in which the linkages between the
determined by titrating 25.0 ml with 0.1 M silver nitrate using dextrose units are almost entirely of the α -1→6 type.
potassium chromate solution as indicator and deducting the
Dextran 110 Injection contains not less than 5.5 per cent and
theoretical value of the sulphated ash present due to the sodium
not more than 6.5 per cent w/v of dextrans.
chloride.
Description. An almost colourless, slightly viscous solution.
Sulphated ash.
Tests
Foreign protein. Inject 0.5 ml on three occasions at intervals
of 2 days into the peritoneal cavity of each of six healthy pH (2.4.24). 3.5 to 6.5 for solutions in Dextrose Injection; 5.0 to
guinea-pigs, weighing not less than 250 g, which have not 7.0 for solutions in Sodium Chloride Injection.
previously been treated with any material, which will interfere Molecular size. For solutions in Dextrose Injection, before
with the test. Inject 0.2 ml intravenously into each of the three proceeding with tests A and B, add 4 volumes of ethanol
393
(95 per cent), centrifuge and dissolve the residue in a volume Ethanol. Distil 100 ml, collect the first 45 ml of distillate and
of Sodium Chloride Injection sufficient to restore the original dilute to 50 ml with water. Mix 10 ml of 0.0167 M potassium
volume. dichromate and 10 ml of sulphuric acid in a stoppered boiling
A (2.4.28). Determine the viscosity ratios by Method A, using tube, immediately add 5 ml of the distillate, mix, stopper the
size C U-tube viscometer at 37°, of solutions in saline solution tube, and allow to stand for 5 minutes. Transfer to a 500-ml
containing about 2.0, 1.0, 0.5 and 0.25 per cent w/v of dextrans, flask, dilute to about 300 ml with carbon dioxide-free water,
accurately determined. For each solution, plot (viscosity ratio add 2 g of potassium iodide and 1 ml of a 10 per cent w/v
- 1.00)/concentration (in per cent w/v) against concentration solution of potassium thiocyanate, allow to stand for
(in per cent w/v). The intercept on the viscosity ratio axis of 5 minutes and titrate the liberated iodine with 0.1 M sodium
the straight line joining the points represents the intrinsic thiosulphate using starch solution, added towards the end
viscosity; the intrinsic viscosity is 0.27 to 0.32. of the titration, as indicator. Repeat the determination
beginning at the words “Mix 10 ml of 0.0167M potassium
B. Place 100 ml in each of five stoppered flasks and adjust the dichromate..” but using 5 ml of water in place of 5 ml of the
temperature to 25.0° ± 0.1°. With precautions to maintain this distillate. The difference between the titrations is not more
temperature, add slowly with continuous stirring sufficient than 4.2 ml.
ethanol to produce a faint cloudiness (about 45 ml is usually
required). Add 0.5, 1.0, 1.5, 2.0 and 2.5 ml of ethanol to the Heavy metals (2.3.13). To 4.0 ml add 5 ml of dilute acetic acid
separate flasks, stopper the flasks and immerse in a water- and sufficient water to produce 25.0 ml. The resulting solution
bath at about 35° with occasional shaking until clear solutions complies with the limit test for heavy metals, Method A
are obtained. Transfer the flasks to a water-bath maintained at (5 ppm).
25.0° ± 0.1° and allow to stand overnight or until two clear Nitrogen (2.3.30). Determine by Method B, using 50 ml. For
liquid phases are formed. Reject the supernatant liquids, solutions in Dextrose Injection, use 30 ml of nitrogen-free
dissolve separately the syrupy residues in sufficient saline sulphuric acid. For solutions in Sodium Chloride Injection
solution to produce 25.0 ml, remove the ethanol by use 20 ml of nitrogen-free sulphuric acid.
evaporation at a pressure of about 2 kPa, dilute to 25.0 ml with
Not more than 0.35 ml of 0.05 M sulphuric acid is required.
water and determine the optical rotation (2.4.22). From the
optical rotations calculate the amount of dextrans precipitated Sulphated ash. Titrate 25 ml with 0.1 M silver nitrate using
as described in the Assay. Choose that fraction containing as potassium chromate solution as indicator. Deduct the
nearly as possible but not more than 10 per cent of the dextrans theoretical value of the sulphated ash due to the sodium
present in the injection and determine its intrinsic viscosity chloride present.
by the method described under test A; the intrinsic viscosity 1 ml of 0.1 M silver nitrate is equivalent to 0.007102 g of
is not more than 0.40. sulphated ash (0.05 per cent w/v)
Content of dextrose. For solutions in Dextrose Injection, Foreign protein. Inject 0.5 ml on three occasions at intervals
between 4.5 and 5.5 per cent w/v determined by the following of 2 days into the peritoneal cavity of each of six healthy
method. Dilute 15.0 ml to 50.0 ml with water. To 5.0 ml in a guinea-pigs weighing not less than 250 g that have not
stoppered flask add 25 ml of a buffer solution containing previously been treated with any material that will interfere
14.3 per cent w/v of sodium carbonate and 4.0 per cent w/v of with the test. Inject 0.2 ml intravenously into each of the three
potassium iodide and 25.0 ml of 0.05 M iodine. Stopper the guinea-pigs 14 days after the first intra-peritoneal injection,
flask and allow to stand for exactly 30 minutes at 20°, add 30 ml and into each of the other three guinea-pigs 21 days after the
of dilute hydrochloric acid and titrate immediately with first intra-peritoneal injection. Observe the guinea-pigs for
0.1 M sodium thiosulphate. Repeat the operation beginning 30 minutes after each intravenous injection and again 24 hours
at the words “add 25 ml of a buffer solution...” but using 5 ml later; the animals exhibit no signs of anaphylaxis such as
of water in place of 5 ml of the preparation under examination. coughing, bristling of hair or respiratory distress.
The difference between the titrations represents the amount
of iodine required to oxidise the dextrose. Bacterial endotoxins (2.2.3). Not more than 1.25 Endotoxin
Units per ml.
1 ml of 0.05 M iodine is equivalent to a 0.00901 g of dextrose.
Acetone. To 10 ml add sufficient ammonium sulphate to give
Preparations (Infusions).
a saturated solution, add 1 ml of sodium nitroprusside solution
and 5 ml of strong ammonia solution, and allow to stand for Assay. For solutions in Dextrose Injection — Add a drop of
10 minutes. Any purple colour produced is not more intense dilute ammonia solution to the required volume and determine
than that produced by treating in the same manner 10 ml of a the optical rotation (2.4.22). Calculate the content of dextrans
0.02 per cent v/v solution of acetone. from the following expression 0.5076(α - 0.528D), where α is
394
IP 2007 DEXTROMETHORPHAN HYDROBROMIDE
the observed angular rotation and D the content of dextrose Ethanol-soluble substances. Not more than 1 per cent,
per cent w/v, determined in the test for Content of dextrose. determined by the following method. Boil under a reflux
For solutions in Sodium Chloride Injection — Measure the condenser 1 g with 20 ml of ethanol (95 per cent) for 5 minutes
optical rotation (2.4.22), and multiply the value obtained by and filter while hot. Evaporate 10 ml of the filtrate on a water-
0.5076. bath, dry the residue at 105° and weigh.
Protein. Not more than 0.5 per cent, determined by the
Storage. Store at a temperature not exceeding 30°. The injection
following method. Carry out Method A for the determination
should not be exposed to undue fluctuations of temperature.
of nitrogen (2.3.30), using 5 g, accurately weighed, and 30 ml
Labelling. The label states (1) the strength as the percentage of nitrogen-free sulphuric acid. Calculate the content of
w/v of dextrans; (2) the name of the solvent; (3) the strain of protein by multiplying the percentage of nitrogen in the
Leuconostoc mesenteroides used; (4) that the injection should substance under examination by 6.25.
not be used if it is cloudy or if a deposit is present.
Reducing substances. Not more than 10 per cent, calculated
as dextrose, C6H12O6, determined by the following method.
Weigh accurately a quantity containing 2 g of the dried
Dextrin substance, add 100 ml of water, shake for 30 minutes, dilute to
Dextrin is starch partially hydrolysed by heat with or without 200.0 ml with water and filter. To 10 ml of cupri-tartaric solution
the aid of suitable acids and buffers. add 20.0 ml of the filtrate, mix and heat at a rate such that the
solution is brought to boil in 3 minutes. Boil for a further
Description. A white or pale yellow powder; odour, slight and 2 minutes and cool quickly. Add 5 ml of a 30 per cent w/v
characteristic. solution of potassium iodide and 10 ml of 1 M sulphuric acid,
mix and titrate immediately with 0.1 M sodium thiosulphate
Identification using starch solution, added towards the end of the titration,
A. Microscopic characteristics - Granules have similar as indicator. Repeat the procedure using 20.0 ml of a 0.1 per
appearance to the starch from which the dextrin has been cent w/v solution of dextrose in place of the filtrate beginning
prepared. In dextrin prepared from maize starch many of the at the words “To 10 ml of...”. Carry out a blank titration using
granules show concentric striations and in dextrin prepared 20 ml of water in place of 20.0 ml of the sample filtrate. The
from potato starch concentric striations are not clearly visible; titre obtained with the sample filtrate is not greater than the
the hilum may be bicleft and some of the granules may be titre obtained with the dextrose solution.
distorted. Ash (2.3.19). Not more than 1 per cent, determined on 1.0 g.
B. Boil 1 g in 50 ml of water, cool. To 5 ml of the cloudy Loss on drying (2.4.19). Not more than 12 per cent, determined
suspension, add a drop of iodine solution and mix; a purple on 1.0 g by drying in an oven at 110°.
colour is produced.
C. To 5 ml of the suspension produced in test B add 2 ml of
2 M sodium hydroxide, mix, add dropwise with shaking 0.5 ml
of cupric sulphate solution and boil; a red precipitate is
produced. Dextromethorphan Hydrobromide
Tests H3CO
Acidity. Add 10 g to 100 ml of ethanol (70 per cent), previously
neutralised to phenolphthalein solution, shake for 1 hour,
filter and titrate 50 ml of the filtrate with 0.1 M sodium hydroxide
using phenolphthalein solution as indicator. Not more than N ,HBr, H2O
H CH3
1 ml of 0.1 M sodium hydroxide is required to change the
colour of the solution.
C18H25NO,HBr,H2O Mol. Wt. 370.3
heavy metals, Method A (40 ppm). Dextromethorphan Hydrobromide is ent-3-methoxy-9a-
Chlorides (2.3.12). Dissolve 2.5 g in 50 ml of boiling water, methylmorphinan hydrobromide monohydrate.
cool, dilute to 100 ml with water and filter. 5 ml of the filtrate Dextromethorphan Hydrobromide contains not less than
diluted to 15 ml complies with the limit test for chlorides 99.0 per cent and not more than 101.0 per cent of C18H25NO,HBr
(0.2 per cent). calculated on the anhydrous basis.
395
DEXTROMETHORPHAN HYDROBROMIDE SYRUP IP 2007
Description. An almost white crystalline powder. Reference solution (c). A 0.25 per cent w/v solution of
dextromethorphan hydrobromide RS in methanol.
Identification
Test A may be omitted if tests B, C and D are carried out. Tests dry the plate in a current of air, spray with potassium
B and C may be omitted if tests A and D are carried out. iodobismuthate solution until spots appear and then
A. Determine by infrared absorption spectrophotometry (2.4.6). immediately spray with hydrogen peroxide solution (10 vol).
Compare the spectrum with that obtained with Any secondary spot in the chromatogram obtained with test
dextromethorphan hydrobromide RS or with the reference solution (a) is not more intense than the spot in the
spectrum of dextromethorphan hydrobromide. chromatogram obtained with reference solution (a) and not
more than one such spot is more intense than the spot in the
B. When examined in the range 230 nm to 360 nm (2.4.7), a chromatogram obtained with reference solution (b).
an absorption maximum only at about 278 nm. Sulphated ash (2.3.18). Not more than 0.1 per cent.
C. In the test for Related substances, the principal spot in the Water (2.3.43). 4.0 to 5.5 per cent, determined on 0.2 g.
chromatogram obtained with test solution (b) corresponds to Assay. Weigh accurately about 0.3 g and dissolve in 20 ml of
that in the chromatogram obtained with reference solution (c). ethanol (95 per cent) and titrate with 0.1 M sodium hydroxide,
D. Gives the reaction of bromides (2.3.1). determining the end-point potentiometrically (2.4.25).
Tests 1 ml of 0.1 M sodium hydroxide is equivalent to 0.03523 g of

C18H25NO,HBr.
(95 per cent) is clear (2.4.1), and colourless (2.4.1).
Acidity or alkalinity. Dissolve 0.4 g in carbon dioxide-free
water with gentle heat, cool and dilute to 20 ml with the same
solvent. Add 0.1 ml of methyl red solution and 0.2 ml of Dextromethorphan Hydrobromide
0.01 M sodium hydroxide. The solution is yellow and not
more than 0.4 ml of 0.01 M hydrochloric acid is required to Syrup
change the colour to red. Dextromethorphan Hydrobromide Syrup is a solution of
Specific optical rotation (2.4.22). +28.0° to +30.0°, determined Dextromethorphan Hydrobromide in a suitable flavoured
in a 2.0 per cent w/v solution in 0.1 M hydrochloric acid. vehicle.
N,N-Dimethylaniline. Dissolve 0.5 g in 20 ml water with the Dextromethorphan Hydrobromide Syrup contains not less
help of gentle heat on a water-bath, cool and add 2 ml of 2 M than 95.0 per cent and not more than 105.0 per cent of the
acetic acid, 1 ml of a 1 per cent w/v solution of sodium nitrite stated amount of dextromethorphan hydrobromide,
and sufficient water to produce 25 ml. The resulting solution C18H25NO,HBr,H2O.
is not more intensely coloured than that obtained by treating
at the same time and in the same manner a solution containing Identification
5 µg of N,N-dimethylaniline in 20 ml of water.
A. To 50 ml, add 20 ml of water, 5 ml of 2.5 M sodium hydroxide
Related substances. Determine by thin-layer chromatography and extract with three quantities, each of 40 ml of hexane,
(2.4.17), coating the plate with silica gel G. collect the hexane layer and filter through anhydrous sodium
Mobile phase. A mixture of 55 volumes of toluene, 20 volumes sulphate placed over absorbent cotton wetted with hexane.
of ethyl acetate, 13 volumes of methanol, 10 volumes of Evaporate the combined extracts at 50° under nitrogen to
dichloromethane and 2 volumes of strong ammonia solution. dryness, dissolve and dilute the residue in 10 ml of chloroform;
the solution is dextrorotatory (2.4.22). Retain the chloroform
Test solution (a). Dissolve 0. 5 g of the substance under
solution for test B.
B. Evaporate the chloroform solution obtained from test A on
a water-bath to dryness, dissolve the residue in 2 ml of 1 M
methanol.
sulphuric acid and add 1 ml of a solution prepared freshly by
Reference solution (a). Dilute 10 ml of test solution (b) to dissolving 700 mg of mercuric nitrate in 4 ml of water, adding
100 ml with methanol. 100 mg of sodium nitrate, mixing and filtering; the solution
Reference solution (b). Dilute 25 ml of reference solution (a) gives no colour, but after heating, a yellow to red colour
to 50 ml with methanol. develops in about 15 minutes.
396
IP 2007 DEXTROSE
Tests Acidity or alkalinity. Dissolve 6.0 g in 25 ml of carbon dioxide-

free water and add 0.3 ml of phenolphthalein solution. The
Other tests. Complies with the tests stated under Oral Liquids. solution is colourless and not more than 0.15 ml of 0.1 M
Assay. Determine by liquid chromatography (2.4.14). sodium hydroxide is required to change the colour of the
Test solution. Dilute a volume of the syrup containing about solution to pink.
10 mg of Dextromethorphan Hydrobromide to 100.0 ml with Specific optical rotation (2.4.22). +52.5° to +53.3°, determined
water. in a solution prepared by dissolving 10.0 g in 80 ml of water,
Reference solution. A 0.01 per cent w/v solution of adding 0.2 ml of 5 M ammonia, mixing well, allowing to stand
dextromethorphan hydrobromide RS. for 30 minutes and diluting to 100.0 ml with water.
Chromatographic system Arsenic (2.3.10). Dissolve 10.0 g in 50 ml of water and add
– a stainless steel column 25 cm x 4.6 mm, packed with 10 ml of stannated hydrochloric acid AsT. The resulting
octadecylsilyl silica gel (5 µm), solution complies with the limit test for arsenic (1 ppm).
– mobile phase: a filtered and degassed solution of Heavy metals (2.3.13). A solution prepared by dissolving 4.0 g
0.007 M ammonium nitrate in a mixture of 70 volumes in 10 ml of water, 2 ml of dilute acetic acid and sufficient
of acetonitrile and 30 volumes of water adjusted to water to produce 25 ml, complies with the limit test for heavy
pH 3.4 with glacial acetic acid, metals, Method A (5 ppm).
– flow rate. 1 ml per minute, Chlorides (2.3.12). 20 ml of a 10.0 per cent w/v solution
– spectrophotometer set at 280 nm, (solution A) complies with the limit test for chlorides (125 ppm).
Sulphates (2.3.17). 7.5 ml of solution A diluted to 15 ml with
Calculate the content of C18H25NO,HBr,H2O. distilled water complies with the limit test for sulphates
Storage. Store protected from light. (200 ppm).
Sulphite. Dissolve 5.0 g in 40 ml of water, add 2.0 ml of 0.1 M
sodium hydroxide and dilute to 50.0 ml with water. To 10.0 ml
Dextrose of the solution add 1 ml of a 31 per cent w/v solution of
hydrochloric acid, 2.0 ml of decolorised magenta solution
Glucose; D-Glucose and 2.0 ml of a 0.5 per cent v/v solution of formaldehyde
HO solution. Allow to stand for 30 minutes and measure the
O 583 nm (2.4.7). The absorbance is not more than that of a
OH OH standard prepared in the following manner. Dissolve 76 mg of
HO sodium metabisulphite in sufficient water to produce 50.0 ml,
OH
dilute 5,0 ml of this solution to 100.0 ml and to 3.0 ml of resulting
C6H12O6 Mol. Wt. 180.2 (anhydrous) solution add 4.0 ml of 0.1 M sodium hydroxide and dilute to
C6H12O6,H2O Mol. Wt. 198.2 (monohydrate) 100.0 ml with water. Immediately treat 10.0 ml of the resulting
solution in the same manner as the test solution beginning at
Dextrose is D-(+)-glucopyranose or D-(+)-glucopyranose the words “add 1 ml of a 31 per cent w/v solution.....”. Use as
monohydrate. the blank for both measurements a solution prepared in the
Description. A white crystalline powder. same manner using 10 ml of water.
Barium. To 10 ml of solution A add 1 ml of 1 M sulphuric acid.
Identification Examine exactly after 1 hour; any opalescence in the solution
A. When heated, it melts, swells up and burns, and an odour is not more intense than that in a mixture of 10 ml of solution A
of burnt sugar is perceptible. and 1 ml of water.
B. Dissolve 0.1 g in 10 ml of water, add 3 ml of potassium Foreign sugars, soluble starch and dextrins. Boil 1.0 g in
cupri-tartrate solution; the solution is blue and clear. Heat to 30 ml of ethanol (90 per cent) to dissolve. The appearance of
boiling; a copious red precipitate is formed. the solution does not change on cooling.
Sulphated ash (2.3.18). Not more than 0.1 per cent determined
Tests
by the following method. Dissolve 5.0 g in 5 ml of water, add
Appearance and odour of solution. Dissolve 10.0 g in 15 ml of 2 ml of sulphuric acid, evaporate to dryness and ignite to
water. The solution is clear (2.4.1), not more intensely coloured constant weight. If necessary, repeat the heating with the
than reference solution BYS7 (2.4.1), and is odourless. sulphuric acid.
397
DEXTROSE INJECTION IP 2007
Water (2.3.43). Not more than 1.0 per cent (anhydrous form) Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Units
and 7.0 to 9.5 per cent (monohydrate), determined on 0.5 g. per ml for preparations containing 5 per cent w/v or less of
Dextrose intended for use in the manufacture of parenteral Dextrose. Dilute injections containing more than 5 per cent
preparations complies with the following additional w/v of Dextrose with sufficient water BET so as to contain
requirement. 5 per cent w/v of Dextose.
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Units Other tests. Complies with the tests stated under Parenteral
per ml of a 5 per cent w/v solution in water for injections. Preparations (Infusions).
Storage. Store protected from moisture. Assay. To an accurately measured volume containing between
2 g and 5 g of Dextrose, add 0.2 ml of 5 M ammonia and
Labelling. The label states (1) whether the material is the
sufficient water to produce 100.0 ml. Mix well, allow to stand
monohydrate or is in the anhydrous form; (2) whether or not
for 30 minutes and determine the optical rotation in a 2-dm
the contents are intended for use in the manufacture of
tube (2.4.22). The observed rotation in degrees multiplied by
parenteral preparations.
0.9477 represents the weight, in g, of dextrose, C6H12O6, in the
volume taken for assay.
Dextrose Injection Storage. Store in single dose containers at a temperature not
exceeding 30°.
Dextrose Intravenous Infusion; Glucose Intravenous
Labelling. The label states (1) the strength as the percentage
Infusion w/v of anhydrous dextrose, C6H12O6; (2) that the injection
Dextrose Injection is a sterile solution of Dextrose in Water for should not be used if it contains visible solid particles.
Injection.
Dextrose Injection contains not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of Diazepam
anhydrous dextrose, C6H12O6.
CH3
Description. A clear, colourless solution. Solutions containing O
20.0 per cent w/v or more of Dextrose may be not more than N
faintly straw-coloured.
Cl N
Identification
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
the solution is blue and clear. Heat to boiling; a copious red
precipitate is formed.
B. The solution prepared as directed in the Assay is C16H13ClN2O Mol. Wt. 284.7
dextrorotatory. Diazepam is 7-chloro-1,3-dihydro-1-methyl-5-phenyl-1,4-
Tests benzodiazepin-2-one.
Diazepam contains not less than 98.5 per cent and not more
pH (2.4.24). 3.5 to 6.5, determined in a solution diluted, if
than 101.0 per cent of C16H13ClN2O, calculated on the dried
necessary, with water for injections to contain not more than
basis.
the equivalent of 5 per cent w/v of Dextrose and to which 0.30
ml of a saturated solution of potassium chloride has been Description. A white or almost white to pale yellow, crystalline
added for each 100 ml of solution. powder; odourless or almost odourless.
5-Hydroxymethylfurfural and related substances. Dilute a Identification
volume containing 1.0 g of Dextrose to 250.0 ml with water.
Absorbance of the resulting solution at the maximum at about Test A may be omitted if tests B, C and D are carried out. Tests
284 nm, not more than 0.25 (2.4.7). B, C and D may be omitted if test A is carried out.
Heavy metals (2.3.13). A solution prepared by evaporating a A. Determine by infrared absorption spectrophotometry (2.4.6).
volume containing 4 g of Dextrose to 10 ml and adding 2 ml of Compare the spectrum with that obtained with diazepam RS
dilute acetic acid and sufficient water to produce 25 ml or with the reference spectrum of diazepam.
complies with the limit test for heavy metals, Method A B. Measure the absorbances in subdued light immediately
(5 ppm). after preparation of the solution.
398
IP 2007 DIAZEPAM CAPSULES
When examined in the range 230 nm to 360 nm (2.4.7), a 0.0005 Diazepam Capsules
per cent w/v solution in 0.05 M methanolic sulphuric acid
shows absorption maxima at about 241 nm and 284 nm; Diazepam Capsules contain not less than 92.5 per cent and
absorbance at about 241 nm, about 0.5 and at about 284 nm, not more than 107.5 per cent of the stated amount of diazepam,
about 0.23. C16H13ClN2O.
C. Measure the absorbance in subdued light immediately Identification
after preparation of this solution.
When examined in the range 325 nm to 400 nm (2.4.7), a
0.0025 per cent w/v solution in 0.05 M methanolic sulphuric
acid shows an absorption maximum only at about 366 nm; Mobile phase. A mixture of 100 volumes of chloroform and
absorbance at about 366 nm, between 0.35 and 0.39. 10 volumes of methanol.
D. Determine by the oxygen-flask method (2.3.34), using Test solution. Shake a quantity of the contents of the capsules
20 mg of the substance under examination and 5 ml of dilute with sufficient methanol to produce a solution containing
sodium hydroxide solution as the absorbing liquid. When 0.5 per cent w/v of Diazepam, allow to settle and decant the
the process is complete, acidify the solution with dilute supernatant liquid.
sulphuric acid and boil gently for 2 minutes; the solution Reference solution. A 0.5 per cent w/v solution of diazepam
gives the reactions of chlorides (2.3.1). RS in methanol.
Tests Apply to the plate 2 µl of each solution. After development,
dry the plate in air, spray with a 10 per cent v/v solution of
Related substances and decomposition products. Determine sulphuric acid in ethanol, heat at 105° for 10 minutes and
in subdued light by thin-layer chromatography (2.4.17), coating examine in ultraviolet light at 365 nm. The principal spot in the
the plate with silica gel GF254. chromatogram obtained with the test solution corresponds to
Mobile phase. A mixture of equal volumes of hexane and that in the chromatogram obtained with the reference solution.
ethyl acetate.
B. When examined in the range 230 nm to 360 nm (2.4.7), the
Prepare the following solutions freshly. solution obtained in the Assay shows two absorption maxima
Test solution. Dissolve 1 g of the substance under examination at about 242 nm and 284 nm.
in sufficient acetone to produce 10 ml.
Tests
with acetone. Related substances and decomposition products. Determine
in subdued light by thin-layer chromatography (2.4.17), coating
Any secondary spot in the chromatogram obtained with the Mobile phase. A mixture of equal volumes of hexane and
test solution is not more intense than the spot in the ethyl acetate.
chromatogram obtained with the reference solution. Prepare the following solutions freshly.
Heavy metals (2.3.13). 1.0 g complies with the limit test for Test solution. Shake a quantity of the contents of the capsules
heavy metals, Method B (20 ppm). containing 50 mg of Diazepam with 5 ml of acetone and filter.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Reference solution. Dilute 1 volume of the test solution to
Loss on drying (2.4.19). Not more than 0.5 per cent, determined 50 volumes with acetone.
on 1.0 g by drying in an oven over phosphorus pentoxide at Apply to the plate 20 µl of the test solution and 5 µl of the
60° at a pressure of 1.5 to 2.5 kPa for 4 hours. reference solution. After development, dry the plate in air and
Assay. Weigh accurately about 0.25 g, dissolve in 80 ml of examine in ultraviolet light at 254 nm. Any secondary spot in
anhydrous glacial acetic acid with the aid of heat, if necessary the chromatogram obtained with the test solution is not more
and cool. Titrate with 0.1 M perchloric acid, determining the intense than the spot in the chromatogram obtained with the
end-point potentiometrically (2.4.25). Carry out a blank titration. reference solution.
1 ml of 0.1 M perchloric acid is equivalent to 0.02847 g of Dissolution (2.5.2).
C16H13ClN2O. Apparatus. No. 1
Storage. Store protected from light. Medium. 900 ml of 0.1 M hydrochloric acid.
399
DIAZEPAM INJECTION IP 2007
Speed and time. 100 rpm and 45 minutes. Mobile phase. A mixture of 100 volumes of chloroform and
Withdraw a suitable volume of the medium and filter promptly 10 volumes of methanol.
through a membrane filter disc with an average pore diameter Test solution. Dilute if necessary, an accurately measured
not greater than 1.0 µm. Reject the first few ml of the filtrate volume of the injection with water to produce a solution
and dilute a suitable volume of the filtrate with 0.1 M containing 5 mg of Diazepam per ml.
solution at the maximum at about 242 nm (2.4.7). Calculate the Reference solution. A 0.5 per cent w/v solution of diazepam
content of diazepam, C16H13ClN2O in the medium from the RS in methanol.
absorbance obtained from a solution of known concentration Apply to the plate 2 µl of each solution. After development,
of diazepam RS. dry the plate in air, spray with a 10 per cent v/v solution of
D. Not less than 85 per cent of the stated amount of sulphuric acid in ethanol, heat at 105° for 10 minutes and
C16H13ClN2O. examine in ultraviolet light at 254 nm. The principal spot in the
Uniformity of content. Comply with the test stated under that in the chromatogram obtained with the reference solution.
Capsules using the following method of analysis. Weigh an
intact capsule. Open the capsule without losing any part of B. When examined in the range 230 nm to 360 nm (2.4.7), the
the shell and transfer the contents as completely as possible solution obtained in the Assay shows absorption maxima at
to a 100-ml volumetric flask. Weigh the shell, remove any about 242 and at about 284 nm.
retained contents and reweigh the shell. To the flask add 1 ml
of water, mix and allow to stand for 15 minutes. Add 80 ml of a Tests
0.5 per cent w/v solution of sulphuric acid in methanol, shake
pH (2.4.24). 6.2 to 6.9.
for 15 minutes, add sufficient of the methanolic sulphuric
acid to produce 100.0 ml and filter. Dilute suitably, if necessary Bacterial endotoxins (2.2.3). Not more than 11.6 Endotoxin
and measure the absorbance at the maximum at about 284 nm Units per mg of diazepam.
(2.4.7). Calculate the content of C16H13ClN2O taking 450 as the Other tests. Complies with the tests stated under Parenteral
specific absorbance at 284 nm, making an appropriate Preparations (Injections).
adjustment for any retained capsule content.
Assay. To an accurately measured volume containing 10 mg
Other tests. Comply with the tests stated under Capsules. of Diazepam, add 20 ml of buffer solution pH 7.0 and extract
Assay. Weigh accurately a quantity of the mixed contents of with four quantities, each of 20 ml, of chloroform, passing
20 capsules containing about 10 mg of Diazepam, add 5 ml of each extract through about 5 g of anhydrous sodium sulphate
water and complete the test as described under Uniformity of and dilute the combined chloroform extracts to 100.0 ml with
content beginning at the words “mix and allow to stand for chloroform. Evaporate 10.0 ml of this solution to dryness under
15 minutes....”. Calculate the content of C16H13ClN2O taking nitrogen, add sufficient volume of a 0.5 per cent w/v solution
450 as the specific absorbance at 284 nm. of sulphuric acid in methanol to produce 100.0 ml. Measure
Storage. Store protected from light. about 284 nm (2.4.7). Calculate the content of C16H13ClN2O
Storage. Store in single dose or multiple dose containers
Diazepam Injection
Diazepam Injection is a sterile solution of Diazepam in Water
for Injections or other suitable solvent.
Diazepam Injection contains not less than 90.0 per cent and Diazepam Tablets
not more than 110.0 per cent of the stated amount of diazepam,
Diazepam Tablets contain not less than 92.5 per cent and not
C16H13ClN2O.
more than 107.5 per cent of the stated amount of diazepam,
Description. A clear, colourless or almost colourless solution. C16H13ClN2O.
A. Determine by thin-layer chromatography (2.4.17), coating A. Determine by thin-layer chromatography (2.4.17), coating
the plate with silica gel G. the plate with silica gel G.
400
IP 2007 DIBUTYL PHTHALATE
Mobile phase. A mixture of 100 volumes of chloroform and D. Not less than 85 per cent of the stated amount of
10 volumes of methanol. C16H13ClN2O.
Test solution. Shake a quantity of the powdered tablets with Uniformity of content. Comply with the test stated under
sufficient methanol to produce a solution containing 0.5 per Tablets.
cent w/v of Diazepam, allow to settle and decant the supernatant Powder one tablet, add 1 ml of water, mix and allow to stand
liquid. for 15 minutes. Add 80 ml of a 0.5 per cent w/v solution of
Reference solution. A 0.5 per cent w/v solution of diazepam sulphuric acid in methanol, shake for 15 minutes, add
RS in methanol. sufficient of the methanolic sulphuric acid to produce 100.0 ml
and filter. Dilute suitably, if necessary and measure the
absorbance at the maximum at about 284 nm (2.4.7). Calculate
the content of C16H13ClN2O in the tablet taking 450 as the
sulphuric acid in ethanol, heat at 105° for 10 minutes and
chromatogram obtained with the test solution corresponds to Other tests. Comply with the tests stated under Tablets.
B. When examined in the range 230 to 360 nm (2.4.7), the quantity of the powder containing about 10 mg of Diazepam,
solution obtained in the Assay shows absorption maxima at add 5 ml of water and complete the test as described under
about 242 and at about 284 nm. Uniformity of content beginning at the words “mix and allow
to stand for 15 minutes....”. Calculate the content of
Tests C16H13ClN2O taking 450 as the specific absorbance at 284 nm.
Related substances and decomposition products. Determine Storage. Store protected from light.
in subdued light by thin-layer chromatography (2.4.17), coating
Mobile phase. A mixture of equal volumes of hexane and
ethyl acetate. Dibutyl Phthalate
Test solution. Prepare freshly by shaking a quantity of the
powdered tablets containing 50 mg of Diazepam with 5 ml of
O
ethanol (95 per cent) and filtering.
Reference solution. Dilute 1 ml of the test solution to 50 ml O CH3
with ethanol (95 per cent). O CH3
Apply to the plate 20 µl of the test solution and 5 µl of the
O
reference solution. After development, dry the plate in air and
the chromatogram obtained with the test solution is not more C16H22O4 Mol. Wt. 278.4
intense than the spot in the chromatogram obtained with the Dibutyl Phthalate is dibutyl benzene-1,2-dicarboxylate.
reference solution.
Dibutyl Phthalate contains not less than 99.0 per cent and not
Dissolution (2.5.2). more than 101.0 per cent of C16H22O4, calculated on the
Apparatus. No 1 anhydrous basis.
Medium. 900 ml of 0.1 M hydrochloric acid. Description. A clear, colourless or very slightly yellow, oily
Speed and time. 100 rpm and 45 minutes. liquid.
through a membrane filter disc with an average pore diameter Identification
not greater than 1.0 µm. Reject the first few ml of the filtrate Test A may be omitted if tests B, C, D and E are carried out.
and dilute a suitable volume of the filtrate with 0.1 M Tests B, C and D may be omitted if tests A and E are carried
hydrochloric acid. Measure the absorbance of the resulting out.
content of diazepam,C16H13ClN2O in the medium from the A. Determine by infrared absorption spectrophotometry (2.4.6).
absorbance obtained from a solution of known concentration Compare the spectrum with that obtained with dibutyl
of diazepam RS. phthalate RS.
401
DICLOFENAC SODIUM IP 2007
B. Determine by thin-layer chromatography (2.4.17), coating Diclofenac Sodium

Mobile phase. A mixture of 70 volumes of ether and 30 volumes
COONa
of n-heptane. Cl H
Test solution. Dissolve 50 mg of the substance under N
examination in sufficient ether to produce 10 ml.
Reference solution. Dissolve 50 mg of dibutyl phthalate RS Cl
in sufficient ether to produce 10 ml.
Apply to the plate 10 µl of each solution. After development, C14H10Cl2NNaO2 Mol. Wt. 318.1
dry the plate in air and examine in ultraviolet light at 254 nm. Diclofenac Sodium is sodium 2-[(2,6-dichlorophenyl)-
The principal spot in the chromatogram obtained with the test amino]phenylacetate.
Diclofenac Sodium contains not less than 98.5 per cent and
not more than 101.0 per cent of C14H10Cl2NNaO2, calculated on
C. To about 0.1 ml add 0.25 ml of sulphuric acid and 50 mg of the dried basis.
resorcinol, heat in a water-bath for 5 minutes, allow to cool
and add 10 ml of water and 1 ml of 10 M sodium hydroxide; Description. A white to slightly yellowish crystalline powder;
the solution becomes yellow or brownish yellow and shows a slightly hygroscopic.
green fluorescence.
Identification
D. Relative density (2.4.29). 1.043 to 1.048.
E. Refractive index (2.4.27). 1.490 to 1.495, determined at 20°.
Compare the spectrum with that obtained with diclofenac
Tests sodium RS or with the reference spectrum of diclofenac
sodium.
Appearance of solution. The liquid under examination is clear
B. To 1 ml of a 0.4 per cent w/v solution in methanol add 1 ml
(2.4.1), and not more intensely coloured than reference solution
of nitric acid; a dark red colour develops.
YS6 (2.4.1).
C. In the test for Related substances, the principal peak in the
Acidity. Dissolve 20.0 g in 50 ml of ethanol (95 per cent)
previously neutralised to phenolphthalein solution and add
the peak in the chromatogram obtained with the reference
0.2 ml of phenolphthalein solution. Not more than 0.5 ml of
solution.
0.1 M sodium hydroxide is required to change the colour of
the solution. C. A 1 per cent w/v solution gives the reaction of sodium salts
(2.3.1).
Water (2.3.43). Not more than 0.2 per cent, determined on Tests
10.0 g.
Assay. Weigh accurately about 1.5 g into a hard-glass flask, methanol is clear (2.4.1), and not more intensely coloured
dissolve in 5 ml of ethanol (95 per cent), previously boiled than reference solution BYS6 (2.4.1).
thoroughly to expel carbon dioxide and neutralised to
phenolphthalein solution. Neutralise any free acid in the pH (2.4.24). 6.5 to 8.5, determined on a 1.0 per cent w/v solution.
solution with 0.1 M ethanolic potassium hydroxide using Light absorption (2.4.7). Absorbance of a 5.0 per cent w/v
0.2 ml of phenolphthalein solution as indicator. Add 25.0 ml solution in methanol at about 440 nm, not more than 0.050.
of 0.5 M ethanolic potassium hydroxide and boil under a
reflux condenser on a water-bath for 1 hour. Add 20 ml of Related substances. Determine by liquid chromatography
water and titrate the excess of alkali with 0.5 M hydrochloric (2.4.14).
acid using a further 0.2 ml of phenolphthalein solution as Test solution. Dissolve 50 mg of the substance under
indicator. Repeat the operation without the substance under examination in methanol and dilute to 50 ml with the same
examination. The difference between the titrations represents solvent.
the alkali required to saponify the ester.
1 ml of 0.5 M ethanolic potassium hydroxide is equivalent to with methanol. Dilute 1 ml of this solution to 10 ml with
0.06959 g of C16H22O4. methanol.
402
IP 2007 DICLOFENAC TABLETS
Chromatographic system Mobile phase. A mixture of 90 volumes of chloroform,

– a stainless steel column 25 cm x 4.6 mm, packed with 5 volumes of acetone and 5 volumes of formic acid in a
end-capped octylsilyl silica gel (5 µm), saturated chamber.
– mobile phase: a mixture of 34 volumes of a solution Test solution. Dilute a suitable volume of the injection
containing 0.5 g per litre of phosphoric acid and 0.8 g containing 25 mg of Diclofenac Sodium to 10 ml with methanol.
per litre of sodium dihydrogen phosphate adjusted to
pH 2.5 with phosphoric acid, and 66 volumes of Reference solution. A 0.25 per cent w/v solution of diclofenac
methanol, sodium RS in methanol.
– flow rate. 1 ml per minute, Apply to the plate 2 µl of each solution. After development,
– spectrophotometer set at 254 nm, dry the plate in a current of warm air and examine in ultraviolet
– a 20 µl loop injector. light at 254 nm. Alternatively, spray with a 0.5 per cent w/v
Inject the test solution and the reference solution. In the solution of potassium dichromate in sulphuric acid (20 per
chromatogram obtained with the test solution: the area of any cent). By both methods of visualisation, the principal spot in
peak other than the principal peak is not greater than the area the chromatogram obtained with the test solution corresponds
of the principal peak in the chromatogram obtained with the to that in the chromatogram obtained with the reference
reference solution (0.2 per cent); the sum of the areas of all solution.
peaks other than the principal peak is not greater than
2.5 times that of the principal peak in the chromatogram Tests
obtained with the reference solution (0.5 per cent). Ignore any pH (2.4.24). 8.1 to 9.0.
peak with an area less than 0.25 times the area of the principal
peak in the chromatogram obtained with the reference solution. Other tests. Complies with the tests stated under Parenteral
heavy metals, Method B (10 ppm). Assay. Determine by liquid chromatography (2.4.14).
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Test solution. Dilute a suitable volume of the injection
on 1.0 g by drying in an oven at 105° for 3 hours. containing 25 mg of Diclofenac Sodium to 10.0 ml with the
mobile phase.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Reference solution. A 0.25 per cent w/v solution of diclofenac
acid, determining the end-point potentiometrically (2.4.25). sodium RS in the mobile phase.
Carry out a blank titration. Chromatographic system
1 ml of 0.1 M perchloric acid is equivalent to 0.03181 g of – a stainless steel column 12.5 cm x 4.6 mm, packed with
C14H10Cl2NNaO2. octylsilyl silica gel (5 µm),
Storage. Store protected from light. 40 volumes of 0.1 M sodium acetate solution,
Diclofenac Injection – a 10 µl loop injector.
Diclofenac Sodium Injection Inject alternately the test solution and the reference solution
and record the chromatograms for 2.5 times the retention time
Diclofenac Injection is a sterile solution of Diclofenac Sodium
of the principal peak. If necessary adjust the concentration of
in Water for Injections. It may contain Propylene Glycol, Benzyl
methanol in the mobile phase to obtain the resolution of the
Alcohol and sufficient Sodium Hydroxide to adjust the pH of
peak due to diclofenac sodium.
the solution.
Calculate the content of C14H10Cl2NNaO2 in the injection.
Diclofenac Injection contains not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of diclofenac
sodium, C14H10Cl2NNaO2.
Diclofenac Tablets
Description. A clear, colourless to yellowish liquid.
Diclofenac Sodium Tablets
Identification
Diclofenac Tablets contain not less than 90.0 per cent and not
Determine by thin-layer chromatography (2.4.17), coating the more than 110.0 per cent of the stated amount of diclofenac
plate with silica gel GF254. sodium, C14H10Cl2NNaO2. The tablets may be enteric-coated.
403
DICYCLOMINE HYDROCHLORIDE IP 2007
Identification Dicyclomine Hydrochloride contains not less than 99.0 per

Determine by thin-layer chromatography (2.4.17), coating the cent and not more than 101.0 per cent of C19H35NO2,HCl,
plate with silica gel 60 F254 or using a precoated silica gel calculated on the dried basis.
60 F254 plate. Description. A white or almost white, crystalline powder;
odourless or almost odourless.
Mobile phase. A mixture of 100 volumes of toluene, 10 volumes
of hexane and 10 volumes of anhydrous formic acid. Identification
A. Dissolve a suitable quantity in acetone and evaporate to
containing 50 mg of Diclofenac Sodium with 5 ml of methanol,
dryness. The residue complies with the following test.
centrifuge and use the supernatant liquid.
Reference solution. A 1 per cent w/v solution of diclofenac
Compare the spectrum with that obtained with dicyclomine
sodium RS in methanol.
Apply separately to the plate 1 µl of each solution. After dicyclomine hydrochloride.
development, dry the plate in a current of warm air and examine
B. To 3 ml of a 0.1 per cent w/v solution of sodium dodecyl
in ultraviolet light at 254 nm. Alternatively, spray the plate
sulphate, add 5 ml of chloroform and 0.05 ml of a 0.25 per cent
with a 0.5 per cent w/v solution of potassium dichromate in
w/v solution of methylene blue, mix gently and allow to
sulphuric acid (20 per cent). By both methods of
separate; the chloroform layer is blue. Add 20 mg of the
visualisation, the principal spot in the chromatogram obtained
substance under examination dissolved in 2 ml of water, mix
with the test solution corresponds to that in the chromatogram
gently and allow to separate; the aqueous layer is blue and
the chloroform layer is colourless.
Tests C. Dissolve 10 mg in 5 ml of water and add 0.2 ml of 2 M nitric
acid and 0.5 ml of silver nitrate solution; a white precipitate is
produced.
quantity of the powder containing about 50 mg of Diclofenac Tests
Sodium, shake with 60 ml of methanol in a 200-ml volumetric Related substances. Determine by thin-layer chromatography
flask and dilute to volume with methanol. Dilute 5.0 ml of this (2.4.17), coating the plate with silica gel G..
solution to 100.0 ml with methanol and measure the absorbance
of the resulting solution at the maximum at about 285 nm (2.4.7). Mobile phase. A mixture of 50 volumes of 1-propanol,
Calculate the content of C14H10Cl2NNaO2 from the absorbance 30 volumes of ethyl acetate, 15 volumes of water and 5 volumes
obtained by repeating the procedure using diclofenac sodium of strong ammonia solution.
RS in place of the substance under examination. Test solution. Dissolve 0.5 g of the substance under
Storage. Store protected from light. examination in 10 ml of methanol.
with methanol and mix. To 2 ml of this solution add sufficient
methanol to produce 100 ml.
Dicyclomine Hydrochloride Apply to the plate 10 µl of each solution. After development,
Dicycloverine Hydrochloride
CH3 intense than the spot in the chromatogram obtained with the
O reference solution.
N CH3 , HCl
O Sulphated ash (2.3.18). Not more than 0.1 per cent.
C19H35NO2,HCl Mol. Wt. 345.9
Dicyclomine Hydrochloride is 2-diethylaminoethyl- acetate solution. Titrate with 0.1 M perchloric acid, using
bicyclohexyl-1-carboxylate hydrochloride. crystal violet solution as indicator. Carry out a blank titration.
404
IP 2007 DICYCLOMINE ORAL SOLUTION
1 ml of 0.1 M perchloric acid is equivalent to 0.03460 g of of sodium hydroxide, diluting to 500 ml with water and
C19H35NO2, HCl. 30 volumes of acetonitrile,
Dicyclomine Injection Inject the reference solution. The test is not valid unless the
tailing factor for the analyte peak is not more than 2.0, and the
Dicyclomine Hydrochloride Injection
Dicyclomine Injection is a sterile, isotonic solution of than 2.0 per cent.
Dicyclomine Hydrochloride in Water for Injections.
Dicyclomine Injection contains not less than 93.0 per cent
Calculate the content of C19H35NO2,HCl in the injection.
dicyclomine hydrochloride, C19H35NO2,HCl. Storage. Store protected from light, in single dose or multiple-
dose containers.
Identification
A. To a volume containing 0.1 g of Dicyclomine Hydrochloride
add 10 ml of water and 1 ml of hydrochloric acid, shake with Dicyclomine Oral Solution
25 ml of ether and allow to separate. Extract the aqueous layer
with 30 ml of chloroform, wash the extract with two quantities, Dicyclomine Hydrochloride Oral Solution; Dicycloverine
each of 10 ml, of water and filter the chloroform solution through Hydrochloride Oral Solution
anhydrous sodium sulphate. Evaporate the filtrate to dryness. Dicyclomine Oral Solution is a solution of Dicyclomine
The residue complies with the following test. Hydrochloride in a suitable flavoured vehicle.
Determine by infrared absorption spectrophotometry (2.4.6). Dicyclomine Oral Solution contains not less than 90.0 per
Compare the spectrum with that obtained with dicyclomine cent and not more than 110.0 per cent of the stated amount of
hydrochloride RS treated in the same manner. dicyclomine hydrochloride, C19H35NO2,HCl.
B. In the Assay the principal peak in the chromatogram
obtained with the test solution corresponds to the peak in the Identification
chromatogram obtained with the reference solution. A. To a volume containing 0.1 g of Dicyclomine Hydrochloride
add 10 ml of water and 1 ml of hydrochloric acid, shake with
Tests
30 ml of ether and allow to separate. Extract the aqueous layer
Bacterial endotoxins (2.2.3). Not more than 17.2 Endotoxin with 30 ml of chloroform, wash the extract with two quantities,
Unit per mg of Dicyclomine Hydrochloride. each of 10 ml, of water and filter the chloroform solution through
anhydrous sodium sulphate. Evaporate the filtrate to dryness,
recrystallise the residue from hot acetone and dry at 105° for
30 minutes. The residue complies with the following test.
Solvent mixture. 1 volume of 0.04 M phosphate buffer, pH 7.5 Compare the spectrum with that obtained with dicyclomine
and 1 volume of acetonitrile. hydrochloride RS or with the reference spectrum of
Test solution. Dilute a volume containing about 20 mg of dicyclomine hydrochloride.
Dicyclomine Hydrochloride to 50.0 ml with the solvent mixture. B. Acidify the oral solution with 2 M nitric acid and add silver
Reference solution. A 0.04 per cent w/v solution of dicyclomine nitrate solution; a white precipitate is produced.
hydrochloride RS in the solvent mixture.
Tests
– a stainless steel column 15 cm x 4.6 mm, packed with Other tests. Complies with the tests stated under Oral Liquids.
octylsilane chemically bonded to porous silica (5 µm), Assay. Weigh accurately a quantity containing about 5 mg of
– mobile phase: a mixture of 70 volumes of 0.02 M Dicyclomine Hydrochloride add 5 ml of sulphuric acid
phosphate buffer pH 7.5 prepared by dissolving 2.72 g (10 per cent v/v) and 2 ml of 0.02 M potassium permanganate,
of monobasic potassium phosphate in 450 ml of water, mix, allow to stand, add 20 ml of water and 20 ml of chloroform
adjusting the pH to 7.5 ± 0.1 with 10 per cent w/v solution to the decolorised solution and titrate with 0.001 M sodium
405
DICYCLOMINE TABLETS IP 2007
dodecyl sulphate, using 1 ml of dimethyl yellow solution as quantities, each of 20 ml, of chloroform, shake with anhydrous
indicator. sodium sulphate, filter, evaporate the filtrate to dryness and
1 ml of 0.001 M sodium dodecyl sulphate is equivalent to dissolve the residue in 4 ml of chloroform.
0.0003460 g of C19H35NO2,HCl. Reference solution. Dilute 1 volume of the test solution to
500 volumes with chloroform.
Determine the weight per ml of the oral solution (2.4.29), and
calculate the content of C19H35NO2,HCl, weight in volume. Apply to the plate 10 µl of each solution. After development,
Storage. Store protected from light. iodobismuthate solution. Any secondary spot in the
Dicyclomine Tablets reference solution.
Dicyclomine Hydrochloride Tablets; Dicycloverine Other tests. Comply with the tests stated under Tablets.
Hydrochloride Tablets Assay. Weigh and powder 20 tablets. Weigh accurately a
Dicyclomine Tablets contain not less than 92.5 per cent and quantity of the powder containing about 30 mg of Dicyclomine
not more than 107.5 per cent of the stated amount of Hydrochloride, add 20 ml of water and shake. Add 10 ml of
dicyclomine hydrochloride, C19H35NO2,HCl. 1 M sulphuric acid, 1 ml of dimethyl yellow solution and
40 ml of chloroform, shake and titrate with 0.004 M sodium
Identification dodecyl sulphate, shaking vigorously and allowing the layers
to separate after each addition, until a permanent orange-pink
colour is produced in the chloroform layer.
of Dicyclomine Hydrochloride with 20 ml of chloroform, filter,
evaporate the filtrate to dryness, recrystallise the residue from 1 ml of 0.004 M sodium dodecyl sulphate is equivalent to
hot acetone and dry at 105° for 4 hours. The residue complies 0.001384 g of C19H35NO2,HCl.
with the following test. Storage. Store protected from light.
Compare the spectrum with that obtained with dicyclomine
hydrochloride RS or with the reference spectrum of Didanosine
dicyclomine hydrochloride.
O
B. To 3 ml of a 0.1 per cent w/v solution of sodium dodecyl
sulphate, add 5 ml of chloroform and 0.05 ml of a 0.25 per cent HN N
w/v solution of methylene blue, mix gently and allow to
separate; the chloroform layer is blue. Add a quantity of the N N
powdered tablets containing 20 mg of Dicyclomine O
Hydrochloride dispersed in 2 ml of water, mix gently and allow HO
to separate; the aqueous layer is blue and the chloroform
layer is colourless.
C10H12N4O3 Mol. Wt. 236.2
C. Shake a quantity of the powdered tablets containing 10 mg
Didanosine is 2′,3′−dideoxyinosine.
of Dicyclomine Hydrochloride with 5 ml of water and 0.2 ml of
2 M nitric acid, filter and add 0.5 ml of silver nitrate solution Didanosine contains not less than 98.0 per cent and not more
to the filtrate; a white precipitate is produced. than 102.0 per cent of C10H12N4O3, calculated on the dried
basis.
Tests
Description. A white or almost white crystalline powder.
(2.4.17), coating the plate with silica gel G.. Identification
Mobile phase. A mixture of 50 volumes of 1-propanol, A. Determine by infrared absorption spectrophotometry (2.4.6).
30 volumes of ethyl acetate, 15 volumes of water and 5 volumes Compare the spectrum with that obtained with didanosine RS
of strong ammonia solution. or with the reference spectrum of didanosine.
Test solution. Shake a quantity of the powdered tablets B. In the Assay, the principal peak in the chromatogram
containing 0.2 g of Dicyclomine Hydrochloride with 8 ml of obtained with the test solution corresponds to the peak in the
water and 2 ml of strong ammonia solution, extract with two chromatogram obtained with the reference solution.
406
IP 2007 DIDANOSINE CAPSULES
Tests Didanosine Capsules

Specific optical rotation (2.4.22). –24.0° to –28.0°, determined Didanosine Capsules contain enteric-coated granules of
in a 1.0 per cent w/v solution in water. Didanosine.
Related substances. Determine by liquid chromatography Didanosine Capsules contain not less than 90.0 per cent and
(2.4.14). not more than 110.0 per cent of the stated amount of
Test solution. Dissolve 0.1 g of the substance under didanosine, C10H12N4O3.
Identification
substance under examination in the mobile phase. In the Assay, the principal peak in the chromatogram obtained
octadecylsilyl silica gel (5 µm), Tests
– mobile phase: a filtered and degassed mixture of
6 volumes of acetonitrile and 94 volumes of water, Dissolution (2.5.2).
– flow rate. 1 ml per minute, A. Apparatus No. 2
– spectrophotometer set at 254 nm, Medium. 1000 ml of 0.1 M hydrochloric acid.
– a 20 µl loop injector. Speed and time. 100 rpm and 120 minutes.
Inject the test solution and the reference solution. In the Determine by liquid chromatography (2.4.14).
Test solution. At the end of the test period dissolve all the
peak other than the principal peak is not greater than half of
granules from the basket in 750 ml of buffer solution pH 7.5
the area of the principal peak in the chromatogram obtained
prepared by dissolving 1.41 g of disodium hydrogen
with the reference solution (0.5 per cent) and the sum of the
orthophosphate anhydrous in 1000 ml of water, adjusting the
areas of all such peaks is not greater than the area of the
pH to 7.5 with orthophosphoric acid and filtering, and dilute
principal peak in the chromatogram obtained with the reference
to 1000 ml with the buffer solution. Dilute suitably to get a
solution (1.0 per cent).
solution containing about 0.005 per cent w/v of didanosine.
didanosine RS in the buffer solution.
Loss on drying (2.4.19). Not more that 0.5 per cent, determined – a stainless steel column 15 cm x 4.6 mm, packed with
on 1.0 g by drying in an oven at 105° for 3 hours. octadecylsilane bonded porous silica (5µm),
Assay. Determine by liquid chromatography (2.4.14) as given – mobile phase: a mixture of 950 volumes of buffer solution
under the test for Related substances using the following pH 7.5 and 50 volumes of acetonitrile,
solutions. – flow rate. 1.5 ml per minute.
Test solution. Dilute 5.0 ml of a 0.1 per cent w/v solution of the – a 10 µl loop injector.
substance under examination in water to 100.0 ml with the
mobile phase. Inject the reference solution. The test is not valid unless the
column efficiency is not less than 3000 theoretical plates the
Reference solution. Dilute 5.0 ml of a 0.1 per cent w/v solution tailing factor is not more than 1.5 and the relative standard
of didanosine RS in water to 100.0 ml with the mobile phase. deviation of replicate injections is not more than 2.0 per cent.
Inject the reference solution. The test is not valid unless the Inject the test solution and the reference solution.
column efficiency determined from the didanosine peak is not
less than 5000 theoretical plates, the tailing factor is not more Calculate the percentage of C10H12N4O3 released in the acid
than 1.5 and the relative standard deviation for replicate medium by subtracting the content of C10H12N4O3 in the test
injections is not more than 2.0 per cent. solution from the total content of didanosine, C10H12N4O3
determined in the Assay.
Not more than 10 per cent of the stated amount of C10H12N4O3
Calculate the content of C10H12N4O3. is dissolved in 120 minutes.
Storage. Store protected from light. B. Apparatus No. 2
407
DIDANOSINE TABLETS IP 2007
Medium. 1000 ml of a buffer solution prepared by mixing 250 Inject reference solution (a). The test is not valid unless the
ml of 0.2 M tribasic sodium phosphate buffer and 750 ml of column efficiency is not less than 4500 theoretical plates and
0.1 M hydrochloric acid and adjusting the pH to 6.8 with 2 M the tailing factor is not more than 2.0.
hydrochloric acid or 2 M sodium hydroxide. Inject the test solution and the reference solution (b). In the
Speed and time. 100 rpm and 45 minutes. chromatogram obtained with the test solution, the area of any
Run for 120 minutes at 100 rpm using the medium given in secondary peak is not more than 4 times the area of the peak in
method A. At the end of this period discard the medium from the chromatogram obtained with the reference solution (b)
each vessel without losing any of the granules and fill the (4.0 per cent) and the sum of all the secondary peaks is not
empty vessel with the dissolution medium preheated to 37º. more than 5.5 times the area of the peak in the chromatogram
After running the apparatus for 45 minutes, withdraw a suitable obtained with the reference solution (5.5 per cent),
volume of the medium and dilute to get a concentration of Other tests. Comply with the tests stated under Capsules.
about 0.005 per cent w/v of didanosine in the dissolution
medium. Assay. Determine by liquid chromatography (2.4.14).
Determine by liquid chromatography (2.4.14). Test solution. Weigh accurately a quantity of the contents of
the capsules containing 50 mg of Didanosine, dissolve in 100.0
Test solution. The solution obtained in the manner described ml of the buffer solution pH 7.5 and filter. Dilute 5.0 ml of the
above. solution to 50.0 ml with the buffer solution pH 7.5.
didanosine RS in the dissolution medium.
didanosine RS in buffer solution pH 7.5.
Use the chromatographic system described in test A.
Inject the reference solution. The test is not valid unless the – a stainless steel column 15 cm x 4.6 mm packed with
column efficiency is not less than 3000 theoretical plates, the octadecylsilane bonded to porous silica (5µm),
tailing factor is not more than 1.5 and the relative standard – mobile phase: a mixture of 950 volumes of buffer solution
deviation of replicate injections is not more than 2.0 per cent. pH 7.5 and 50 volumes of acetonitrile,
Inject the test solution and the reference solution. – flow rate. 1.5 ml per minute.
C10H12N4O3.
column efficiency is not less than 3000 theoretical plates the
(2.4.14).
NOTE — Prepare the solutions immediately before use. deviation of replicate injections is not more than 2.0 per cent.
Test solution. Weigh accurately a quantity of the contents of Inject the test solution and the reference solution.
the capsules containing 100 mg of Didanosine, dissolve in
Calculate the content of C10H12N4O3 in the capsules.
100 ml of mobile phase and filter.
exceeding 30º.
didanosine RS in the mobile phase.
100 ml with the mobile phase. Didanosine Tablets
Didanosine Tablets contain not less than 90.0 per cent and
octadecylsilane bonded to porous silica (5µm), (such
didanosine, C10H12N4O3. The tablets may contain permitted
as Lichrospher RP18e),
flavouring agents.
prepared by dissolving 1.15 g of ammonium dihydrogen Identification
orthophosphate in 1000 ml of water, and 5 volumes of
acetonitrile, adjust the pH to 6.8 with triethylamine A. Shake a quantity of the powdered tablets containing 0.1 g
and filter, of Didanosine with 80 ml of water, dilute to 100 ml with water
– flow rate. 1 ml per minute. and filter. Dilute 5 ml of the filtrate to 100 ml with water. When
– spectrophotometer set at 254 nm, examined in the range 220 nm to 350 nm (2.4.7), the resulting
– a 20 µl loop injector. solution shows an absorption maximum only at about 250 nm.
408
IP 2007 DIENOESTROL
B. In the Assay, the principal peak in the chromatogram not be more than 6.0 per cent when calculated by percentage
obtained with the test solution corresponds to the peak in the area normalisation.
chromatogram obtained with the reference solution. Other tests. Comply with the tests stated under Tablets.
Tests Assay. Determine by liquid chromatography (2.4.14).
Related substances. Determine by liquid chromatography Test solution. Weigh accurately a quantity of the powdered
(2.4.14). tablets containing about 100 mg of Didanosine and transfer to
a 100-ml volumetric flask. Add about 50 ml of buffer solution
Test solution. Weigh accurately a quantity of the powdered pH 7.0, mix with the aid of ultrasound for 10 minutes, dilute to
tablets containing about 50 mg of Didanosine and transfer to volume with the same solvent, mix and filter through a
a 50-ml volumetric flask. Add about 25 ml of buffer solution membrane filter disc with an average pore diameter not greater
pH 7.0, and mix with the aid of ultrasound for 5 minutes, dilute than 0.45 µm.
to volume with the same solvent, mix and filter.
Reference solution. A 0.1 per cent w/v solution of didanosine
Reference solution. Weigh accurately about 50 mg didanosine RS in buffer solution pH 7.0. Filter through a membrane filter
RS and transfer to a 50-ml volumetric flask. Dissolve in about disc with an average pore diameter not greater than 0.45 µm.
25 ml of buffer solution pH 7.0 and dilute to volume with the
same solvent. Dilute 5.0 ml of this solution to 50.0 ml with the
same solvent. Dilute further 5.0 ml to 50.0 ml with the same
octadecylsilyl silica gel (5 µm)( such as Kromasil C18),
solvent and filter through a membrane filter disc with an average
pore diameter not greater than 0.45 µm.
5 volumes of acetonitrile and 95 volumes of a buffer
Chromatographic system solution prepared by dissolving 1.42 g of disodium
– a stainless steel column 15 cm x 4.6 mm, packed with hydrogen phosphate in 1000 ml of water, adjusting the
octadecylsilyl silica gel (5 µm)( such as Kromasil C18), pH to 7.5 ± 0.05 with dilute phosphoric acid,
– mobile phase: filtered and degassed gradient mixtures – flow rate. 1.5 ml per minute,
of acetonitrile and buffer solution pH 7.0 prepared by – spectrophotometer set at 245 nm,
dissolving 1.42 g of disodium hydrogen phosphate and – a 5 µl loop injector.
6.8 g of tetrabutylammonium hydrogen sulphate in 1000 Inject the reference solution and record the chromatogram for
ml of water, adjusting the pH of the solution to twice the retention time of didanosine. The test is not valid
7.0 ± 0.05 with sodium hydroxide solution, unless the column efficiency determined from the didanosine
– flow rate. 1.5 ml per minute, peak is not less than 4500 theoretical plates, the tailing factor
– a linear gradient programme using the conditions given is not more than 1.5 and the relative standard deviation for
below, replicate injections is not more than 1.0 per cent.
– a 5 µl loop injector. Inject alternately the test solution and the reference solution.
Time Buffer (pH 7.0) Acetonitrile Coment Calculate the content of C10H12N4O3 in the tablets.
(in min.) (per cent v/v) (per cent v/v) Storage. Store protected from light.
0–8 100 → 0 0 isocratic Labelling. The label states that the tablets should be chewed
20 – 25 70 →30 30 linear gradient before swallowing.
26 – 35 100 → 0 0 re-equilibrium
Inject the reference solution. The test is not valid unless the Dienoestrol
column efficiency determined from the didanosine peak is not
less than 3000 theoretical plates and the tailing factor is not Dienestrol
more than 1.5.
H3C H OH
Inject separately the buffer and test solution. Examine the
chromatogram obtained with the buffer solution for any
extraneous peaks and ignore the corresponding peaks
observed in the chromatogram obtained with the test solution. HO H CH3
Any secondary peak observed in the chromatogram obtained
with the test solution should not be more than 5.0 per cent C18H18O2 Mol. Wt. 266.3
and the sum of the areas of all the secondary peaks should Dienoestrol is (E,E)-4,4′ -[bis(ethylidene)ethylene]-diphenol.
409
DIENOESTROL TABLETS IP 2007
Dienoestrol contains not less than 98.5 per cent and not more is not valid unless the chromatogram obtained with reference
than 101.5 per cent of C18H18O2, calculated on the dried basis. solution (c) shows at least two clearly separated spots having
Description. A white or almost white, crystalline powder. approximately the same intensity.
Identification
Test A may be omitted if tests B, C and D are carried out. Tests on 1.0 g by drying in an oven at 105°.
B and C may be omitted if tests A and D are carried out. Assay. Weigh accurately about 25 mg and dissolve in sufficient
A. Determine by infrared absorption spectrophotometry (2.4.6). ethanol to produce 100.0 ml. To 5.0 ml of this solution add
Compare the spectrum with that obtained with dienoestrol 10 ml of ethanol, dilute with 0.1 M sodium hydroxide to
RS. 250.0 ml and measure the absorbance of the resulting solution
of C18H18O2 from the absorbance obtained by repeating the
procedure using dienoestrol RS in place of the substance
under examination.
C. Heat a mixture of about 1 mg in 5 ml of glacial acetic acid
and 1 ml of a 1 per cent w/v solution of bromine in glacial
acetic acid in a water-bath for 2 minutes. To 0.5 ml of the
solution in a dry test tube add 0.5 ml of ethanol, mix and add
10 ml of water; a reddish-violet colour is produced. Add 5 ml Dienoestrol Tablets
of chloroform, shake vigorously and allow to separate; the
chloroform layer is red and the aqueous layer is almost Dienestrol Tablets
colourless. Dienoestrol Tablets contain not less than 90.0 per cent and
D. Dissolve 0.5 mg in 0.2 ml of glacial acetic acid, add 1 ml of not more than 110.0 per cent of the stated amount of
phosphoric acid and heat on a water-bath for 3 minutes; a dienoestrol, C18H18O2.
reddish-violet colour is produced.
Identification
Tests
A. Extract a quantity of the powdered tablets containing about
Related substances. Determine by thin-layer chromatography 15 mg of Dienoestrol with ether and filter; evaporate the filtrate
(2.4.17), coating the plate with silica gel G. to dryness. Reserve a portion of the residue for test C. Heat a
mixture of about 1 mg of the residue in 5 ml of glacial acetic
acid and 1 ml of a 1 per cent w/v solution of bromine in
10 volumes of diethylamine.
glacial acetic acid in a water-bath for 2 minutes. To 0.5 ml of
Test solution (a). Dissolve 0.5 g of the substance under the solution in a dry test tube add 0.5 ml of ethanol, mix and
examination in 5 ml of ethanol (95 per cent). add 10 ml of water; a reddish-violet colour is produced. Add
Test solution (b). Dilute 5 ml of test solution (a) to 100 ml with 5 ml of chloroform, shake vigorously and allow to separate;
ethanol (95 per cent). the chloroform layer is red and the aqueous layer is almost
colourless.
dienoestrol RS in ethanol (95 per cent). B. Dissolve 0.5 mg of the residue in 0.2 ml of glacial acetic
acid, add 1 ml of phosphoric acid and heat on a water-bath
Reference solution (b). Dilute 5 ml of reference solution (a) to for 3 minutes; a reddish-violet colour is produced.
50 ml with ethanol (95 per cent).
Reference solution (c). A solution containing 0.25 per cent the plate with silica gel G.
w/v each of dienoestrol RS and stilbestrol RS in ethanol
(95 per cent). Mobile phase. A mixture of 90 volumes of toluene and
10 volumes of diethylamine.
dry the plate in air, spray with ethanolic sulphuric acid Test solution. Shake a quantity of the powdered tablets
(20 per cent v/v) and heat at 120° for 10 minutes. Any containing 2 mg of Dienoestrol with 4 ml of acetone, centrifuge
secondary spot in the chromatogram obtained with test and use the supernatant liquid.
solution (a) is not more intense than the spot in the Reference solution (a). A 0.05 per cent w/v solution of
chromatogram obtained with reference solution (b). The test dienoestrol RS in acetone.
410
IP 2007 DIETHYLCARBAMAZINE CITRATE
Reference solution (b). A solution containing 0.1 per cent Diethylcarbamazine Citrate is N,N-diethyl-4-
w/v each of dienoestrol RS and stilbestrol RS in acetone. methylpiperazine-1-carboxamide dihydrogen citrate.
Apply to the plate 10 µl of each solution. After development, Diethylcarbamazine Citrate contains not less than 98.0 per
dry the plate in air, spray with ethanolic sulphuric acid cent and not more than 101.0 per cent of C10H21N3O,C6H8O7,
(20 per cent v/v) and heat at 120° for 10 minutes. The principal calculated on the dried basis.
Description. A white, crystalline powder; odourless; slightly
hygroscopic.
reference solution (a). The test is not valid unless the
chromatogram obtained with reference solution (b) shows two Identification
clearly separated spots having approximately the same
intensity. A. Determine by infrared absorption spectrophotometry (2.4.6).
Tests diethylcarbamazine citrate RS or with the reference spectrum
of diethylcarbamazine citrate.
Uniformity of content. Comply with test stated under Tablets.
B. In the test for N,N‘-Dimethylpiperazine and
Powder one tablet and extract with successive quantities of
N-methylpiperazine, the principal spot in the chromatogram
ether until complete extraction is effected. Filter the ether
solution and wash the filter with small quantities of ether.
chromatogram obtained with the reference solution (a).
Evaporate the ether and dissolve the residue in 10 ml of
ethanol and add sufficient 0.1 M sodium hydroxide to C. A 2 per cent w/v solution gives reaction A of citrates (2.3.1).
produce a solution containing 0.0005 per cent w/v of
Dienoestrol. Complete the test as described in the Assay Tests
beginning at the words “Measure the absorbance....”. Appearance of solution. A 10.0 per cent w/v solution is not
Calculate the content of C18H18O2 in the tablet. more opalescent than opalescence standard OS2 (2.4.1), and
Other tests. Comply with the tests stated under Tablets. not more intensely coloured than reference solution BYS6
(2.4.1).
quantity of the powder containing about 10 mg of Dienoestrol N,N’-Dimethylpiperazine and N-methylpiperazine. Determine
and triturate with successive quantities of ether until complete by thin-layer chromatography (2.4.17), coating the plate with
extraction is effected. Filter the ether extracts and wash the silica gel G.
filter with small quantities of ether. Combine the filtrate and Mobile phase. A mixture of 65 volumes of methanol,
washings and remove the ether; dissolve the residue in 30 volumes of 2-butanone and 5 volumes of strong ammonia
sufficient ethanol to produce 50.0 ml. To 5.0 ml of this solution solution.
add 10 ml of ethanol and sufficient 0.1 M sodium hydroxide
to produce 200.0 ml. Measure the absorbance of the resulting
content of C18H18O2 from the absorbance obtained by repeating Reference solution (a). A 5 per cent w/v solution of
the operation using a solution obtained by dissolving 2.5 mg, diethylcarbamazine citrate RS in methanol.
accurately weighed, of dienoestrol RS in 20 ml of ethanol and Reference solution (b). A 0.01 per cent w/v solution of N,N‘-
diluting with sufficient 0.1 M sodium hydroxide to produce dimethylpiperazine in methanol.
500.0 ml.
Storage. Store protected from light. N-methylpiperazine in methanol.
Diethylcarbamazine Citrate phase to rise 12 cm. Dry the plate at 105° and expose it to
iodine vapour for 30 minutes. Any spots corresponding to
O N,N‘-dimethylpiperazine and N-methylpiperazine in the
N N CH3 HO COOH intense than the spots in the chromatogram obtained with
,
N HOOC COOH reference solutions (b) and (c) respectively.
H3C CH3
Heavy metals (2.3.13). 1.0 g dissolved in 20 ml of water, 0.5 ml
C10H21N3O,C6H8O7 Mol. Wt. 391.4 of 0.1 M hydrochloric acid and sufficient water to produce
411
DIETHYLCARBAMAZINE TABLETS IP 2007
25 ml complies with the limit test for heavy metals, Method A Determine by infrared absorption spectrophotometry (2.4.6).
(20 ppm). Compare the spectrum with that obtained with
Sulphated ash (2.3.18). Not more than 0.1 per cent. diethylcarbamazine citrate RS or with the reference spectrum
of diethylcarbamazine citrate.
on 1.0 g by drying in an oven over phosphorus pentoxide at Tests
60° at a pressure of 1.5 to 2.5 kPa for 4 hours.
N,N’-Dimethylpiperazine and N-methylpiperazine. Determine
Assay. Determine by liquid chromatography (2.4.14). by thin-layer chromatography (2.4.17), coating the plate with
Test solution. Weigh accurately about 25 mg of the substance silica gel G.
under examination, dissolve in 20 ml of a 3.124 per cent w/v Mobile phase. A mixture of 65 volumes of methanol,
solution of potassium dihydrogen phosphate, dilute to 30 volumes of 2-butanone and 5 volumes of strong ammonia
25.0 ml with the same solvent, mix well and filter. Dilute 5.0 ml solution.
of the filtrate to 50.0 ml with the potassium dihydrogen
phosphate solution. Test solution. Dissolve 0.5 g of the substance under
diethylcarbamazine citrate RS in a 3.124 per cent w/v solution Reference solution (a). A 5 per cent w/v solution of
of potassium dihydrogen phosphate. diethylcarbamazine citrate RS in methanol.
Chromatographic system Reference solution (b). A 0.01 per cent w/v solution of
– a stainless steel column 30 cm x 3.9 mm, packed with N,N‘- dimethylpiperazine in methanol.
octadecylsilyl silica gel (5 µm), Reference solution (c). A 0.01 per cent w/v solution of
– mobile phase: a mixture of 100 volumes of methanol and N-methylpiperazine in methanol.
900 volumes of a 1 per cent solution of potassium
dihydrogen phosphate, Apply to the plate 10 µl of each solution. Allow the mobile
– flow rate. 2.5 ml per minute, phase to rise 12 cm. Dry the plate at 105° and expose it to
– spectrophotometer set at 220 nm, iodine vapour for 30 minutes. Any spots corresponding to
– a 20 µl loop injector. N,N‘-dimethylpiperazine and N-methylpiperazine in the
Inject alternately suitable volumes of the test solution and intense than the spots in the chromatogram obtained with
reference solution. The test is not valid unless the relative reference solutions (b) and (c) respectively.
standard deviation of the peak areas of diethylcarbamazine in
replicate injections is not more than 2.0 per cent. Dissolution (2.5.2).
Calculate the content of C10H21N3O,C6H8O7. Apparatus. No 1

Diethylcarbamazine Tablets not greater than 1.0 µm, rejecting the first few ml of the filtrate.
Diethylcarbamazine Citrate Tablets Dilute the filtrate, if necessary, with an equal volume of a
6.248 per cent w/v solution of potassium dihydrogen
Diethylcarbamazine Tablets contain not less than 92.5 per cent
phosphate. Carry out the determination as described in the
Assay. Calculate the content of C10H21N3O,C6H8O7 using a
diethylcarbamazine citrate, C10H21N3O,C6H8O7.
solution of known concentration of diethylcarbamazine
Identification citrate RS in a 3.124 per cent w/v solution of potassium
dihydrogen phosphate.
To a quantity of the powdered tablets containing 0.15 g of
Diethylcarbamazine Citrate add 15 ml of ethanol (95 per cent), D. Not less than 75 per cent of the stated amount of
shake for 5 minutes, filter and evaporate the filtrate to dryness. C10H21N3O,C6H8O7.
To the residue add 10 ml of 2 M sodium hydroxide and extract Other tests. Comply with the tests stated under Tablets.
with three quantities, each of 10 ml, of chloroform. Dry the
combined extracts over anhydrous sodium sulphate, filter and
evaporate the chloroform. The residue complies with the Test solution. Weigh and powder 20 tablets. Weigh accurately
following test. a quantity of the powder containing about 25 mg of
412
IP 2007 DIETHYL PHTHALATE
Diethylcarbamazine Citrate, add 20 ml of a 3.124 per cent w/v Relative density (2.4.29). about 1.010 at 30º,
solution of potassium dihydrogen phosphate and place in an Refractive index (2.4.27). 1.439 to 1.447 at 20º sodium D lines.
ultrasonic bath for 5 minutes. Cool, dilute to 25.0 ml with the
same solvent and filter. Dilute 5.0 ml of the filtrate to 50.0 ml Water (2.3.43). Not more than 0.5 per cent.
with the same solvent. Assay. Determine by gas chromatography (2.4.13).
Reference solution. A 0.01 per cent w/v solution of Internal standard solution. A 3 per cent w/v solution of
diethylcarbamazine citrate RS in a 3.124 per cent w/v solution diethyl sebaeate in acetone.
of potassium dihydrogen phosphate.
Test solution. To 0.2 g of the substance under examination,
Chromatographic system add 10 ml of internal standard solution and dilute to the 100.0
– a stainless steel column 30 cm x 3.9 mm, packed with ml with acetone.
– mobile phase: a mixture of 100 volumes of methanol and Reference solution. To 50 mg of diethyl phenylacetamide
900 volumes of a 1 per cent w/v solution of potassium RS, add 2.5 ml of internal standard solution and dilute to the
dihydrogen phosphate, 25.0 ml with acetone.
– spectrophotometer set at 220 nm, – a glass column 1.2 m x 2 mm packed with 10 per cent OV-
– a 20 µl loop injector. 101 on chromosorb WHP (100-120 mesh)
Inject alternately suitable volumes of the test solution and – temperature :
reference solution. The test is not valid unless the relative column 150º,
standard deviation of the peak areas of diethylcarbamazine in inlet port and detector at 300º,
replicate injections is not more than 2.0 per cent. – flow rate 30 ml per minute of the Nitrogen, 30 ml per
minute of the Hydrogen, 210 ml per minute of the Air.
Calculate the content of C10H21N3O,C6H8O7. in the tablets. Inject 2 µl of the test solution and the reference solution.
Storage. Store protected from moisture. Calculate the content of C12H17NO.
Diethyl Phenyl Acetamide
O Diethyl Phthalate
C12H14O4 Mol. Wt. 222.2
N CH3
Diethyl phthalate is diethyl benzene-1,2-dicarboxylate.
CH3
Diethyl phthalate contains not less than 99.0 per cent and not
more than 101.0 per cent of diethyl phthalate, C12H14O4.
C12H17NO Mol. Wt. 191.3
Description. A clear, oily liquid, colourless or very slightly
Diethyl Phenyl Acetamide is N,N-diethylbenzeneacetamide
yellow.
Diethyl Phenyl Acetamide contains not less than 99.0 per cent
and not more than 101.0 per cent of C12H17NO, calculated on Identification
the anhydrous basis.
Description. A clear to faintly yellow liquid. It shall be free B, C and D may be omitted if test A is carried out.
from suspended matter.
Identification Compare the spectrum with that obtained with diethyl
phthalate RS.
B. Relative density (2.4.29). 1.117 to 1.121.
chromatogram obtained with the reference solution. C. Determine by thin layer chromatography (2.4.17), coating
the plate with silica gel GF 254.
Tests
Mobile phase. A mixture of 30 volumes of heptane and
Boiling point (2.4.8). about 143º, 70 volumes of ether.
413
DIETHYLTOLUAMIDE IP 2007
Test solution. Dissolve 50 mg of the substance under Inject 1 µl of test solution (a). In the chromatogram obtained,
examination in 10 ml of ether. verify that there is no peak with the same retention time as the
Reference solution. Dissolve 50 mg of diethyl phthalate RS internal standard.
in 10 ml of ether. Inject separately 1 µl of test solution (b) and the reference
solution. Continue the chromatography for three times the
Apply to the plate 10 µl of each solution. After development
retention time of diethyl phthalate. From the chromatogram
obtained with the reference solution, calculate the ratio (R) of
the area of the peak due to diethyl phthalate to the area of the
peak due to the internal standard. From the chromatogram
with reference solution.
obtained with test solution (b), calculate the ratio of the sum
D. To about 0.1 ml, add 0.25 ml of sulphuric acid and 50 mg of of the areas of any peaks, other than the principal peak and
resorcinol. Heat on a water-bath for 5 minutes. Allow to cool. the peaks due to the internal standard and the solvent, to the
Add 10 ml of water and 1 ml of strong sodium hydroxide area of the peak due to the internal standard; this ratio is not
solution. The solution becomes yellow or brownish-yellow greater than R (1.0 per cent).
and shows green fluorescence.
Tests 5.0 g.
Appearance. The substance under examination is clear (2.4.1)
on 1.0 g.
and not more intensely coloured than reference solution YS6,
(2.4.1). Assay. Weigh accurately about 0.75 g, dissolve in 25.0 ml of
0.5 M alcoholic potassium hydroxide and add few glass
Acidity. Dissolve 20.0 g in 50 ml of alcohol previously
beads. Boil on a water-bath under a reflux condenser for
neutralised to phenolphthalein solution. Add 0.2 ml of
1 hour. Add 1 ml of phenolphthalein solution and titrate
phenolphthalein solution. Not more than 0.1 ml of 0.1 M
immediately with 0.5 M hydrochloric acid. Carry out a blank
sodium hydroxide is required to change the colour of the
titration.
indicator to pink.
1 ml of 0.5 M alcoholic potassium hydroxide is equivalent to
Related substances. Determine by gas chromatography
0.05556 g of C12H14O4.
(2.4.13).
Internal standard solution. Dissolve 60 mg of naphthalene
in 20 ml of methylene chloride.
Test solution (a). Dissolve 1 g of the substance under
examination in 20 ml of methylene chloride.
Diethyltoluamide
Test solution (b). Dissolve 1 g of the substance under
examination in methylene chloride, add 2.0 ml of the internal Deet
standard solution and dilute to 20 ml with methylene chloride.
Reference solution. To 1 ml of test solution (a) add 10 ml of the O
internal standard solution and dilute to 100 ml with methylene H3C
N CH3
chloride.
Chromatographic system CH3
– a glass column 2.0 m x 2 mm, packed with silanised
diatomaceous earth for gas chromatography (150 µm to C12H17NO Mol. Wt. 191.3
180 µm) impregnated with 3 per cent m/m of Diethyltoluamide is N,N-diethyl-3-toluamide.
polymethylphenylsiloxane,
– temperature : Diethyltoluamide contains not less than 95.0 per cent and not
column 150°, more than 103.0 per cent of C12H17NO, calculated on the
inlet port and detector at 225°, anhydrous basis.
– flow rate. 30 ml per minute of the carrier gas. Description. A colourless or faintly yellow liquid; odourless
Inject 1 µl of the reference solution. The test is not valid unless or almost odourless.
the resolution between the peaks corresponding to CAUTION — Diethyltoluamide is irritant to the eyes and
naphthalene and diethyl phthalate is at least 10. mucous membranes.
414
IP 2007 DIGITOXIN
Identification Digitoxin
O O
diethyltoluamide RS. H3C
B. Heat 2 ml with 25 ml of a 50 per cent v/v solution of H3C H
hydrochloric acid under a reflux condenser for 1 hour. Make
the mixture alkaline with sodium hydroxide solution, cool and H OH
extract with three quantities, each of 30 ml, of ether. Reserve CH3 O
the aqueous layer. Evaporate the ether, dissolve the residue in O H
5 ml of dilute hydrochloric acid, cool to 5°, add 5 ml of sodium
nitrite solution and allow to stand for 10 minutes at 5°. Add O 3
10 ml of water and extract with two quantities, each of 20 ml, of OH
H
ether. Evaporate the ether, add 1 g of phenol to the residue,
cool and add 1 ml of sulphuric acid; an intense green colour
is produced, which becomes red on pouring into water and C41H64O13 Mol. Wt. 764.9
green on making alkaline with dilute sodium hydroxide
Digitoxin is 3β-[(O-2,6-dideoxy-β-D-ribo-hexopyranosyl-
solution.
(1→ 4)-O-2,6-dideoxy-β-D-ribo-hexopyranosyl-
C. Acidify the aqueous layer reserved in test B with dilute (1128W1Ä4)-2,6-dideoxy-β-D-ribo-hexopyranosyl)oxy]-14β-
hydrochloric acid, extract with two quantities, each of 20 ml, hydroxy-5β-card-20(22)-enolide.
of ether and evaporate the ether from the combined extracts.
Digitoxin contains not less than 95.0 per cent and not more
The residue, after drying at 60°, melts at about 108° (2.4.21).
than 103.0 per cent of C41H64O13, calculated on the dried basis.
Tests Description. A white or almost white powder; odourless.
Weight per ml (2.4.29). 0.997g to 1.000 g, determined at 20°.
Identification
Refractive index (2.4.27). 1.520 to 1.524.
Acidity. A solution of 10.0 g dissolved in 50 ml of ethanol B, C and D may be omitted if test A is carried out.
(95 per cent) previously neutralised to phenolphthalein
solution requires not more than 4.0 ml of 0.01 M sodium A. Determine by infrared absorption spectrophotometry 2.4.6).
hydroxide to change the colour of the solution, using Compare the spectrum with that obtained with digitoxin RS.
phenolphthalein solution as indicator. B. In the test for Related substances, the principal spot in the
Sulphated ash (2.3.18). Not more than 0.1 per cent. chromatogram obtained with the test solution corresponds to
1.0 g. C. Dissolve about 1 mg in 2 ml of glacial acetic acid with the
aid of gentle heat, cool and add 0.05 ml of ferric chloride test
Assay. Weigh accurately about 0.3 g, add 7 ml of nitrogen-
solution. Cautiously add 1 ml of sulphuric acid under the two
free sulphuric acid and carry out the determination of nitrogen
liquids without mixing; a brown ring develops at the interface
(2.3.30), using 0.05 M sulphuric acid as the titrant.
which gradually becomes blue and a green colour, finally
1 ml of 0.05 M sulphuric acid is equivalent to 0.01913 g of passes to the upper layer.
C12H17NO.
D. Suspend about 0.5 mg in 0.2 ml of ethanol (60 per cent)
Storage. Store protected from moisture in dry containers. and add 0.1 ml dinitrobenzoic acid solution and 0.1 ml of 2 M
sodium hydroxide; a violet colour develops.
Tests
of equal volumes of chloroform and methanol is clear (2.4.1),
415
DIGITOXIN TABLETS IP 2007
Specific optical rotation (2.4.22). +16.0 ° to +18.5°, determined absorbance of the resulting solution at the maximum at about
at 20° in a 2.5 per cent w/v solution in chloroform. 495 nm (2.4.7), using as the blank a mixture of 5.0 ml of ethanol
(95 per cent) and 3.0 ml of alkaline picric acid solution.
Calculate the content of C41H64O13 from the absorbance
obtained by repeating the operation using digitoxin RS in
Mobile phase. A mixture of 90 volumes of chloroform, place of the substance under examination.
40 volumes of cyclohexane and 15 volumes of methanol.
Storage. Store protected from moisture and light in a refrigerator
Solvent mixture. A mixture of equal volumes of chloroform (2° to 8°).
and methanol.
Digitoxin Tablets
Reference solution (a). A 1.0 per cent w/v solution of digitoxin
RS in the same solvent mixture. Digitoxin Tablets contain not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of digitoxin,
Reference solution (b). Dilute 1 ml of reference solution (a) to C41H64O13.
100 ml with the same solvent mixture.
Reference solution (c). A 0.02 per cent w/v solution of gitoxin Identification
RS in the same solvent mixture.
To a quantity of the powdered tablets containing 250 µg of
Reference solution (d). Dilute 5 ml of reference solution (b) to Digitoxin add 1 ml of glacial acetic acid containing 0.01 per
10 ml with the same solvent mixture. cent w/v of ferric chloride, shake for a few minutes, filter
Reference solution (e). A solution containing 0.5 per cent through sintered-glass and add cautiously 1 ml of sulphuric
w/v of digitoxin RS and 0.01 per cent w/v of gitoxin RS in the acid to the filtrate without mixing; a brown ring free from red
same solvent mixture. colour is produced at the interface which gradually becomes
blue and finally the upper layer acquires an indigo colour.
Apply to the plate 5 µl of each solution and develop the
chromatograms immediately after applying the solutions. After Tests
development, dry the plate in a current of cold air for 5 minutes.
Repeat the development and again dry the plate in a current of Dissolution (2.5.2).
cold air for 5 minutes. Spray with ethanolic sulphuric acid Apparatus. No 2
(10 per cent) and heat at 130° for 15 minutes. Examine the Medium. 600 ml of freshly distilled water
chromatograms in daylight. Any spot in the chromatogram Speed and time. 120 rpm and 60 minutes.
obtained with the test solution corresponding to gitoxin is
not more intense than the spot in the chromatogram obtained Place six tablets in each basket in the test
with reference solution (c). Any other secondary spot in the Withdraw a suitable volume of the medium and filter through
chromatogram obtained with the test solution is not more a membrane filter disc having an average pore diameter not
intense than the spot in the chromatogram obtained with greater than 0.8 µm, rejecting the first 1 ml of the filtrate. Transfer
reference solution (b). The test is not valid unless the 1.0 ml to a 10-ml volumetric flask, add 3.0 ml of a 0.1 per cent
chromatogram obtained with reference solution (e) shows w/v solution of L-ascorbic acid in methanol and 0.2 ml of a
clearly separated spots corresponds to digitoxin and gitoxin 0.009M solution of hydrogen peroxide [prepared by accurately
and the spot in the chromatogram obtained with reference diluting hydrogen peroxide solution (100 vol) that has been
solution (d) is clearly visible. standardised by titration with 0.02 M potassium
permanganate], mix and dilute to volume with hydrochloric
acid. After exactly 30 minutes measure the fluorescence of the
on the residue obtained from the test for Loss on drying.
solution (2.4.5), using an excitation wavelength of about
Loss on drying (2.4.19). Not more than 1.5 per cent, determined 400 nm and an emission wavelength of about 570 nm and
on 0.5 g by drying in an oven at 105° for 2 hours. setting the spectrophotofluorimeter to zero with water and to
Assay. Weigh accurately about 40 mg, dissolve in sufficient 100 with a solution of suitable concentration of digitoxin RS
ethanol (95 per cent) to produce 50.0 ml and dilute 5.0 ml of prepared at the same time and treated in the same manner as
this solution to 100.0 ml with the same solvent. To 5.0 ml of the test solution.
this solution add 3.0 ml of alkaline picric acid solution, allow D. Not less than 75 per cent of the stated amount of digitoxin,
to stand in subdued light for 30 minutes and measure the C41H64O13.
416
IP 2007 DIGOXIN
Uniformity of content. Comply with test stated under Tablets. Digoxin

Test solution. For tablets containing 100 µg of Digitoxin, shake
1 tablet with 15 ml of methanol (50 per cent) for 30 minutes
O O
and dilute to 25.0 ml with the same solvent.
For tablets containing 200 µg of Digitoxin, shake 1 tablet with HO
30 ml of methanol (50 per cent) for 30 minutes and dilute to H3C
50.0 ml with the same solvent.
H3C H
Filter through a suitable membrane filter disc having an average
pore diameter not greater than 0.8 µm, rejecting the first few ml H OH
of the filtrate. Transfer 1.0 ml to a 10-ml volumetric flask, add CH3 O
3.0 ml of a 0.1 per cent w/v solution of L-absorbic acid in O H
methanol and 0.2 ml of a 0.009M solution of hydrogen peroxide
[prepared by accurately diluting hydrogen peroxide solution O 3
(100 vol) that has been standardised by titration with 0.02 M H OH
potassium permanganate], mix and dilute to volume with
hydrochloric acid. After exactly 30 minutes measure the C41H64O14 Mol. Wt. 780.9
fluorescence of the solution (2.4.5), using an excitation
Digoxin is 3β-[(O-2,6-dideoxy-β-D-ribo-hexopyranosyl-
wavelength of about 400 nm and an emission wavelength of
(1→ 4)-O-2,6-dideoxy-β-D-ribo-hexopyranosyl-
about 570 nm and setting the spectrophotofluorimeter to zero
(1128W1Ä4)-2, 6-dideoxy-β-D-ribo-hexopyranosyl)oxy]-
with water. Calculate the content of digitoxin, C41H64O13, from
12b,1428β-dihydroxy-5β-card-20(22)-enolide.
the fluorescence obtained by carrying out the operation
described above at the same time using a 0.0004 per cent w/v Digoxin contains not less than 95.0 per cent and not more
solution of digitoxin RS in methanol (50 per cent) and than 103.0 per cent of C41H64O14, calculated on the dried basis.
beginning at the words “Transfer 1.0 ml to a 10-ml volumetric Description. Colourless crystals or a white or almost white
flask....”. powder.
Identification
a quantity of the powder containing about 1.25 mg of Digitoxin, Test A may be omitted if tests B ,C and D are carried out. Tests
add 3.0 ml of water, swirl to disperse the powder and allow to B, C and D may be omitted if test A is carried out.
stand for 10 minutes, swirling occasionally. Add 25.0 ml of A. Determine by infrared absorption spectrophotometry (2.4.6).
glacial acetic acid, shake for 1 hour and filter, discarding the Compare the spectrum with that obtained with digoxin RS or
first few ml of the filtrate. To 4.0 ml of the filtrate add 1.0 ml of with the reference spectrum of digoxin.
dimethyl sulphoxide, dilute to 25.0 ml with xanthydrol
reagent, mix well and allow to stand in the dark for 4 ½ hours B. In the test for Related substances, the principal spot in the
(solution A). At the same time prepare two further solutions in chromatogram obtained with the test solution corresponds to
the same manner but using for solution B 4.0 ml of digitoxin that in the chromatogram obtained with reference solution (a).
standard solution and for solution C 4.0 ml of a mixture of C. Dissolve about 1 mg in 2 ml of glacial acetic acid with the
25 volumes of glacial acetic acid and 3 volumes of water and aid of gentle heat, cool and add 0.05 ml of ferric chloride test
beginning at the words “add 1.0 ml of dimethyl solution. Cautiously add 1 ml of sulphuric acid under the two
sulphoxide......”. Measure the absorbances of solutions A and liquids without mixing; a brown ring develops at the interface
B at the maximum at about 550 nm (2.4.7), using solution C as which gradually becomes blue and a green colour, finally
the blank. Calculate the content of C 41H64O13 from the passes to the upper layer.
absorbances obtained.
D. Suspend about 0.5 mg in 0.2 ml of ethanol (60 per cent)
Storage. Store protected from light and moisture at a and add 0.1 ml dinitrobenzoic acid solution and 0.1 ml of 2 M
temperature not exceeding 30°. sodium hydroxide; a violet colour develops.
Tests
of equal volumes of dichloromethane and methanol is clear
417
DIGOXIN INJECTION IP 2007
Specific optical rotation (2.4.22). +10.0° to +13.0°, determined Assay. Weigh accurately about 40 mg, dissolve in sufficient
in a 2.0 per cent w/v solution in anhydrous pyridine. ethanol (95 per cent) to produce 50.0 ml and dilute 5.0 ml of
Related substances. Determine by thin-layer chromatography this solution to 100.0 ml with the same solvent. To 5.0 ml of the
(2.4.17), coating the plate with kieselguhr G. Place the dry resulting solution add 3.0 ml of alkaline picric acid solution,
plate in a closed tank containing the necessary quantity of a allow to stand in subdued light for 30 minutes and measure
mixture of 90 volumes of acetone and 10 volumes of formamide the absorbance of the solution at the maximum at about
so that the plate dips about 5 mm into the liquid and allow the 495 nm (2.4.7), using as the blank a mixture of 5.0 ml of
impregnating solvent to ascend at least 15 cm. Remove the ethanol(95 per cent) and 3.0 ml of alkaline picric acid
plate from the tank, allow to stand for 30 minutes and then use solution. Calculate the content of C 41H 64O 14 from the
immediately. absorbance obtained by repeating the operation using digoxin
RS in place of the substance under examination.
50 volumes of xylene and 4 volumes of formamide.
Solvent mixture. A mixture of equal volumes of
dichloromethane and methanol.
Test solution. Dissolve 0.1g of the substance under examination
in 10 ml with solvent mixture. Digoxin Injection
Reference solution (a). A 1.0 per cent w/v solution of digoxin Digoxin Injection is a sterile solution of Digoxin in Water for
RS in the same solvent mixture. Injections and Ethanol or other suitable solvents.
Reference solution (b). Dilute 1 ml of reference solution (a) to Digoxin Injection contains not less than 90.0 per cent and not
50 ml with the same solvent mixture. more than 110.0 per cent of the stated amount of digoxin,
Reference solution (c). A 0.01 per cent w/v solution of digoxin C41H64O14.
Reference solution (d). Dilute 1 ml of reference solution (a) to
Identification
100 ml with the same solvent mixture. Evaporate 2 ml to dryness, dissolve the residue in 1 ml of
Reference solution (e). A 0.02 per cent w/v of gitoxin RS in glacial acetic acid containing 0.01 per cent w/v of ferric
the same solvent mixture. chloride and cautiously add 1 ml of sulphuric acid without
mixing; a brown ring develops at the interface which gradually
Apply to the plate 2 µl of each solution. Allow the mobile becomes blue and finally the upper layer acquires a blue colour.
phase to rise 12 cm. Dry the plate in a current of cold air until
only the lower edge is still moist. Repeat the development and Tests
dry the plate at 115° for 20 minutes. Allow to cool, spray with
a mixture of 15 volumes of a 25 per cent w/v solution of pH (2.4.24). 6.7 to 7.3.
trichloroacetic acid in ethanol (95 per cent) and 1 volume of
freshly prepared 3 per cent w/v solution of chloramine T and
heat at 115° for 5 minutes. Examine in ultraviolet light at
365 nm. Any spots corresponding to digitoxin and gitoxin in Assay. Transfer 20 ml, accurately measured, to a separating
the chromatogram obtained with the test solution are not more funnel containing 10 ml of water. Make alkaline with 5 M
intense than the spots in the chromatograms obtained with ammonia and extract with four quantities, each of 25 ml, of
reference solutions (d) and (e) respectively. Any other chloroform. Wash each extract with the same 10 ml of water.
secondary spot in the chromatogram obtained with the test Evaporate the combined extracts to dryness on a water-bath,
solution is not more intense than the spot in the chromatogram dissolve the residue in 5.0 ml of a mixture of 65 volumes of
obtained with reference solution (b) and not more than one chloroform and 35 volumes of methanol and add 20.0 ml of
such spot is more intense than the spot in the chromatogram glacial acetic acid (solution A). To 5.0 ml of a 0.2 per cent
obtained with reference solution (c). w/v solution of digoxin RS in glacial acetic acid add 10.0 ml
of a mixture of 65 volumes of chloroform and 35 volumes of
methanol and sufficient glacial acetic acid to produce
on the residue obtained from the test for Loss on drying.
50.0 ml (solution B). Dilute 5.0 ml of solution A to 25.0 ml with
Loss on drying (2.4.19). Not more than 1.0 per cent, determined digoxin reagent, mix, allow to stand for 1 hour and measure
on 0.5 g by drying over phosphorus pentoxide at a pressure the absorbance of the resulting solution at about 590 nm,
not exceeding 2.7 kPa. using water as the blank (2.4.7). Calculate the content of
418
IP 2007 DIGOXIN TABLETS
C41H64O14 from the absorbance obtained by treating 5.0 ml of beginning at the words “Dilute 5.0 ml of the filtrate......” and
solution B at the same time and in the same manner. using water as the blank.
Storage. Store protected from light in single dose containers. Storage. Store protected from light at a temperature not
exceeding 30°.
Digoxin Paediatric Solution

Digoxin Tablets
Paediatric Digoxin Elixir
Digoxin Tablets contain not less than 90.0 per cent and not
Digoxin Paediatric Solution is a solution of Digoxin in a suitable more than 110.0 per cent of the stated amount of digoxin,
flavoured vehicle. C41H64O14.
Digoxin Paediatric Solution contains not less than 90.0 per
cent and not more than 110.0 per cent w/v of the stated amount Identification
of digoxin, C41H64O14.
To a quantity of the powdered tablets containing 250 µg of
Digoxin add 1 ml of glacial acetic acid containing 0.01 per
Identification
cent w/v of ferric chloride, shake for a few minutes, filter
Digoxin Paediatric Solution should not be diluted before use through sintered-glass and add cautiously 1 ml of sulphuric
and should be measured with a pipette. acid to the filtrate without mixing; a brown ring free from red
colour is produced at the interface which gradually becomes
Extract a quantity of the solution containing 250 µg of Digoxin
blue and finally the upper layer acquires an indigo colour.
with four quantities, each of 20 ml, of chloroform, washing
each extract with the same 10 ml of water, evaporate the
combined extracts to dryness and dissolve the residue in 1 ml
Tests
of glacial acetic acid containing 0.01 per cent w/v of ferric Dissolution (2.5.2).
chloride. Add cautiously 1 ml of sulphuric acid without
Apparatus. No 2
mixing; a brown ring develops at the interface which gradually
becomes blue and finally the upper layer acquires a blue colour. Medium. 600 ml of freshly distilled water
Tests Place six tablets in each basket in a test
pH (2.4.24). 6.8 to 7.2. Withdraw a suitable volume of the medium and filter through
a membrane filter disc having an average pore diameter not
Other tests. Complies with the tests stated under Oral Liquids.
greater than 0.8 µm, rejecting the first 1 ml of the filtrate. Transfer
Assay. Extract an accurately measured volume containing 1.0 ml to a 10-ml volumetric flask, add 3.0 ml of a 0.1 per cent
about 5 mg of Digoxin with four quantities, each of 25 ml, of w/v solution of L-absorbic acid in methanol and 0.2 ml of a
chloroform, washing each extract with the same 5 ml of water, 0.009M solution of hydrogen peroxide (prepared by accurately
and evaporate the combined extracts to dryness. To the residue diluting hydrogen peroxide solution (100 vol) that has been
add 3 ml of ethanol and carefully evaporate to dryness on a standardised by titration with 0.02 M potassium
water-bath with the aid of a gentle current of air. Repeat the permanganate), mix and dilute to volume with hydrochloric
evaporation using a further 3 ml of ethanol and cool. Dissolve acid. After exactly 2 hours measure the fluorescence of the
the residue in 5.0 ml of a mixture of 65 volumes of chloroform solution (2.4.5), using an excitation wavelength of about
and 35 volumes of methanol, add 20.0 ml of glacial acetic 360 nm and an emission wavelength of about 490 nm and
acid and filter if necessary. Dilute 5.0 ml of the filtrate to setting the spectrophotofluorimeter to zero with water and to
25.0 ml with digoxin reagent, allow to stand for 1 hour and 100 with a solution prepared at the same time as the test solution
measure the absorbance of the resulting solution at the in the following manner. Dilute 2.5 ml of a 0.1 per cent w/v
maximum at about 590 nm (2.4.7). Calculate the content of solution of digoxin RS in ethanol (80 per cent) to 100.0 ml
C41H64O14 from the absorbance obtained by carrying out the with water, dilute the resulting solution further with water to
operation described above at the same time but using a produce a solution containing in 1 ml an amount of digoxin
solution prepared by mixing 5.0 ml of a 0.2 per cent w/v solution equal to one-hundredth of the strength of the tablets under
of digoxin RS in glacial acetic acid with 10.0 ml of a mixture examination, transfer 1.0 ml of the solution to a 10-ml volumetric
of 65 volumes of chloroform and 35 volumes of methanol and flask and carry out the operation described above, beginning
adding sufficient glacial acetic acid to produce 50.0 ml at the words “add 3.0 ml….”.
419
DIIODOHYDROXYQUINOLINE IP 2007
D. Not less than 75 per cent of the stated amount of digoxin, Diiodohydroxyquinoline
C41H64O14.
Iodoquinol
Tablets.
OH
Test solution. For tablets containing 250 µg of Digoxin, place
I N
1 tablet with 10 ml of water at 37°, agitate to disintegrate, add
56 ml of ethanol (95 per cent), shake for 60 minutes and add
sufficient ethanol (80 per cent) to produce 100.0 ml.
For tablets containing 125 µg and 62.5 µg of Digoxin, repeat I
the above procedure by using proportionately smaller
C9H5I2NO Mol. Wt. 396.9
quantities of water, ethanol (95 per cent) and ethanol
(80 per cent). Diiodohydroxyquinoline is 5,7-diiodoquinolin-8-ol.
Filter through a suitable membrane filter disc having an average Diiodohydroxyquinoline contains not less than 97.0 per cent
pore diameter not greater than 0.8 µm, rejecting the first few ml and not more than 100.5 per cent of C9H5I2NO, calculated on
of the filtrate. Transfer 1.0 ml to a 10-ml volumetric flask, add the dried basis.
3.0 ml of a 0.1 per cent w/v solution of L-absorbic acid in Description. A light yellowish to yellowish-brown,
methanol and 0.2 ml of a 0.009M solution of hydrogen peroxide microcrystalline powder; odourless or with a faint odour.
[prepared by accurately diluting hydrogen peroxide solution
(100 vol) that has been standardised by titration with 0.02 M Identification
potassium permanganate], mix and dilute to volume with
hydrochloric acid. After exactly 2 hours measure the A. Determine by infrared absorption spectrophotometry (2.4.6).
fluorescence of the solution (2.4.5), using an excitation Compare the spectrum with that obtained with
wavelength of about 360 nm and an emission wavelength of diiodohydroxyquinoline RS or with the reference spectrum
about 490 nm and setting the spectrophotofluorimeter to zero of diiodohydroxyquinoline.
with water. Calculate the content of digoxin, C41H64O14, from B. Dissolve 10 mg in 100 ml of dioxan and dilute 5 ml to 100 ml
the fluorescence obtained by carrying out the operation with ethanol. When examined in the range 230 nm to 360 nm
described above at the same time using a 0.00025 per cent (2.4.7), the resulting solution shows an absorption maximum
w/v solution of digoxin RS in ethanol (80 per cent) and at about 258 nm; absorbance at about 258 nm, about 0.53.
beginning at the words “Transfer 1.0 ml to a 10-ml volumetric
C. Heat a few crystals with about 1 ml of sulphuric acid; violet
flask....”
vapours of iodine are evolved.
Tests
a quantity of the powder containing about 1.25 mg of Digoxin, Acidity or alkalinity. Shake 0.5 g with 10 ml of water previously
add 3.0 ml of water, swirl to disperse the powder and allow to neutralised to phenolphthalein solution. The solution is
stand for 10 minutes, swirling occasionally. Add 25.0 ml of colourless and not more than 0.1 ml of 0.1 M sodium hydroxide
glacial acetic acid, shake for 1 hour and filter, discarding the is required to change the colour of the solution to pink.
first few ml of the filtrate. To 4.0 ml of the filtrate add 1.0 ml of Free iodine and iodide. Shake 1.0 g with 20 ml of water for
dimethyl sulphoxide, dilute to 25.0 ml with xanthydrol 30 seconds, allow to stand for 5 minutes and filter. To 10 ml of
reagent, mix well and allow to stand in the dark for 4 1/2 hours the filtrate add 1 ml of 1 M sulphuric acid and 2 ml of
(solution A). At the same time prepare two further solutions in chloroform and shake; the chloroform layer does not become
the same manner but using for solution B 4.0 ml of digoxin violet. To the mixture add 5 ml of 1 M sulphuric acid and 1 ml
standard solution and for solution C 4.0 ml of a mixture of of potassium dichromate solution and shake for 15 seconds;
25 volumes of glacial acetic acid and 3 volumes of water and the colour of the chloroform layer does not become more
beginning at the words “add 1.0 ml of dimethyl intense than that produced by diluting 2 ml of a 0.016 per cent
sulphoxide......”. Measure the absorbances of solutions A and w/v solution of potassium iodide to 10 ml with water, adding
B at the maximum at about 545 nm (2.4.7), using solution C as 6 ml of 1 M sulphuric acid, 1 ml of potassium dichromate
the blank. Calculate the content of C 41H64O14 from the solution and 2 ml of chloroform and shaking for 15 seconds.
absorbances obtained.
Related substances. Determine by gas chromatography
Storage. Store protected from light. (2.4.13).
420
IP 2007 DIIODOHYDROXYQUINOLINE TABLETS
Test solution. Add 0.5 ml of N,O-bis (trimethylsilyl)acetamide Diiodohydroxyquinoline Tablets

to 0.5 ml of a solution in pyridine containing 0.4 per cent w/v
of each of 5-chloro-8- hydroxyquinoline, 5,7-dichloro-8- Iodoquinol Tablets
hydroxyquinoline and 5-chloro-7-iodo-8-hydroxyquinoline Diiodohydroxyquinoline Tablets contain not less than
and 0.04 per cent w/v of the substance under examination, 90.0 per cent and not more than 110.0 per cent of the stated
mix, allow to stand for 15 minutes and add 5 ml of a 0.05 per amount of diiodohydroxyquinoline, C9H5I2NO.
cent w/v solution of dibutylphthalate (internal standard) in
hexane. Identification
Reference solution (a). Add 0.5 ml of N,O-bis (trimethylsilyl) A. Triturate a quantity of the powdered tablets containing
acetamide to a mixture of 0.1 g of the substance under about 50 mg of Diiodohydroxyquinoline with 10 ml of carbon
examination and 0.5 ml of pyridine, mix, allow to stand for 15 disulphide, filter and evaporate the solvent. The residue
minutes and add 5 ml of hexane. complies with the following test.
Reference solution (b). Treat a mixture of 0.1 g of the substance Determine by infrared absorption spectrophotometry (2.4.6).
under examination and 0.5 ml of pyridine as described for the Compare the spectrum with that obtained with
diiodohydroxyquinoline RS or with the reference spectrum
test solution.
of diiodohydroxyquinoline.
Chromatographic system B. Shake a quantity of the powdered tablets containing about
– a glass column 1.5m x 4 mm, packed with silanised 10 mg of Diiodohydroxyquinoline with 100 ml of dioxan, filter
diatomaceous support (100 to 120 mesh) coated with and dilute 5 ml of the filtrate to 100 ml with ethanol. When
3 per cent w/w of methyl silicone gum, examined in the range 230 nm to 360 nm (2.4.7), the resulting
– temperature: solution shows an absorption maximum at about 258 nm;
column.190°, absorbance at about 258 nm, about 0.53 (2.4.7).
inlet port and detector. 240°,
Tests
– nitrogen as carrier gas. Soluble iodides. Digest a quantity of the powdered tablets
containing 0.1 g of Diiodohydroxyquinoline with 5 ml of water
In the chromatogram obtained with the test solution the peaks
for 10 minutes, cool and filter. To the filtrate add 1 ml of 3 M
following the solvent peak, in order of emergence, are due to
hydrochloric acid, 0.1 ml of ferric chloride test solution and
(a) 5-chloro-8-hydroxyquinoline, (b) 5,7-dichloro-8-hydroxy-
2 ml of chloroform, shake gently and allow to separate; any
quinoline, (c) the internal standard, (d) 5-chloro-7-iodo-8-
violet colour in the chloroform is not more intense than that in
hydroxyquinoline and (e) diiodohydroxyquinoline. In the a blank to which 1 ml of a 0.02 per cent w/v solution of
chromatogram obtained with reference solution (b) calculate potassium iodide has been added.
the content of 5-chloro-8-hydroxy-quinoline, 5,7-dichloro-8-
hydroxyquinoline and 5-chloro-7-iodo-8-hydroxyquinoline by Disintegration (2.5.1). 30 minutes.
reference to the corresponding peaks in the chromatogram Other tests. Comply with the tests stated under Tablets.
obtained with the test solution. The total content of the named Assay. Weigh and powder 20 tablets. Weigh accurately a
impurities and any other impurities does not exceed 4.0 per quantity of the powder containing about 12 mg of
cent w/w. Diiodohydroxyquinoline and determine by the oxygen-flask
Sulphated ash (2.3.18). Not more than 0.1 per cent. method (2.3.34), using a mixture of 10 ml of water and 2 ml of
1 M sodium hydroxide as the absorbing liquid. When the
Loss on drying (2.4.19). Not more than 0.5 per cent, determined process is complete, add to the flask an excess (5 ml to 10 ml)
on 1.0 g by drying over phosphorus pentoxide at a pressure of acetic bromine solution and allow to stand for 2 minutes.
not exceeding 0.7 kPa for 4 hours. Remove the excess of bromine by the addition of formic acid
Assay. Weigh accurately about 0.3 g and dissolve in 50 ml of (about 0.5 ml to 1 ml). Rinse the sides of the flask with water
anhydrous pyridine. Titrate with 0.1 M tetrabutylammonium and sweep out any bromine vapour above the liquid with a
hydroxide, determining the end-point potentiometrically current of air. Add 1 g of potassium iodide and titrate with
(2.4.25). Carry out a blank titration. 0.02 M sodium thiosulphate using starch solution, added
towards the end of the titration, as the indicator.
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
1 ml of 0.02 M sodium thiosulphate is equivalent to
0.03969 g of C9H5I2NO. 0.0006616 g of C9H5I2NO.
421
DILOXANIDE FUROATE IP 2007
Diloxanide Furoate Apply to the plate 5 µl of each solution. After development,

CH3 Cl test solution is not more intense than the spot in the
N chromatogram obtained with the reference solution.
O Cl
O O
O Loss on drying (2.3.19). Not more than 0.5 per cent, determined
C14H11Cl2NO4 Mol. Wt. 328.2 anhydrous pyridine. Titrate with 0.1 M tetrabutylammonium
Diloxanide Furoate is 4-(N-methyl-2,2-dichloroacetamido) hydroxide, determining the end-point potentiometrically
phenyl 2-furoate. (2.4.25). Carry out a blank titration.
Diloxanide Furoate contains not less than 98.0 per cent and 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
not more than 102.0 per cent of C14H11Cl2NO4, calculated on 0.03282 g of C14H11Cl2NO4.
the dried basis. Storage. Store protected from light.
odourless or almost odourless
Identification
Diloxanide Tablets
Diloxanide Furoate Tablets
Compare the spectrum with that obtained with diloxanide Diloxanide Tablets contain not less than 95.0 per cent and not
furoate RS or with the reference spectrum of diloxanide furoate. more than 105.0 per cent of the stated amount of diloxanide
furoate, C14H11Cl2NO4.
0.001 per cent w/v solution in ethanol (95 per cent) shows an
Identification
absorption maximum only at about 258 nm; absorbance at
about 258 nm, about 0.70. A. Extract a quantity of the powdered tablets containing 0.2 g
C. On 20 mg determine by the oxygen-flask method (2.3.34), of Diloxanide Furoate with 20 ml of chloroform, filter and
using 10 ml of 1 M sodium hydroxide as the absorbing liquid. evaporate the filtrate to dryness. The residue complies with
When the process is complete, acidify the liquid with nitric the following test.
acid and add silver nitrate solution; a white precipitate is Determine by infrared absorption spectrophotometry (2.4.6).
produced. Compare the spectrum with that obtained with diloxanide
furoate RS or with the reference spectrum of diloxanide furoate.
Tests
B. On 20 mg of the residue obtained in test A determine by the
Free acidity. Shake 3.0 g with 50 ml of water, filter and wash oxygen-flask method (2.3.34), using 10 ml of 1 M sodium
the residue with three quantities, each of 20 ml, of water. Titrate hydroxide as the absorbing liquid. When the process is
the combined filtrate and washings with 0.1 M sodium complete, acidify the liquid with nitric acid and add silver
hydroxide using phenolphthalein solution as indicator; not nitrate solution; a white precipitate is produced.
more than 1.3 ml is required. C. The residue obtained in test A melts at 114° to 116° (2.4.21).
(2.4.17), coating the plate with silica gel HF254. Tests
Mobile phase. A mixture of 96 volumes of dichloromethane Related substances. Determine by thin-layer chromatography
and 4 volumes of methanol. (2.4.17), coating the plate with silica gel HF254.
Test solution. Dissolve 0.5 g of the substance under Mobile phase. A mixture of 96 volumes of dichloromethane
examination in 5 ml of chloroform. and 4 volumes of methanol.
Reference solution. Dilute 1 ml of the test solution to 100 ml Test solution. Shake a quantity of the powdered tablets
with chloroform and mix. Dilute 5 ml of the resulting solution containing 0.5 g of Diloxanide Furoate with 5 ml of chloroform,
to 20 ml with chloroform. centrifuge and use the supernatant liquid.
422
IP 2007 DILTIAZEM HYDROCHLORIDE
Reference solution. Dilute 1 ml of the test solution to 100 ml B. In the test for Related substances, the principal peak in the
with chloroform and mix. Dilute 5 ml of the resulting solution chromatogram obtained with the test solution corresponds to
to 20 ml with chloroform. the peak due to diltiazem hydrochloride in the chromatogram
Apply to the plate 5 µl of each solution. After development, obtained with the reference solution.
dry the plate in air and examine in ultraviolet light at 254 nm. C. A 5 per cent w/v solution gives the reactions of chlorides
Any secondary spot in the chromatogram obtained with the (2.3.1).
chromatogram obtained with the reference solution. Tests
Other tests. Comply with the tests stated under Tablets. Specific optical rotation (2.4.22). +110° to +116°, determined
Assay. Weigh and powder 20 tablets. Weigh accurately a in a 1.0 per cent w/v solution.
quantity of the powder containing about 40 mg of Diloxanide Related substances. Determine by liquid chromatography
Furoate, shake with 150 ml of ethanol (95 per cent) for (2.4.14).
30 minutes, add sufficient ethanol (95 per cent) to produce
200.0 ml, mix and filter. Dilute 10.0 ml of the filtrate to 250.0 ml Test solution. Dissolve 0.12g of the substance under
with ethanol (95 per cent) and measure the absorbance of examination in 100 ml of methanol.
the resulting solution at the maximum at about 258 nm (2.4.7). Reference solution. A solution containing 0.12 per cent w/v
Calculate the content of C14H11Cl2NO4 taking 705 as the specific each of diltiazem hydrochloride RS and desacetyl diltiazem
absorbance at 258 nm. hydrochloride RS in methanol.
Storage. Store protected from light. Chromatographic system
Diltiazem Hydrochloride containing 0.116 per cent w/v of d-10-
camphorsulphonic acid in 0.1 M sodium acetate, with
the pH adjusted to 6.2 by the addition of 0.1 M sodium
OCH3
hydroxide, 25 volumes of acetonitrile and 25 volumes
of methanol, filtered and degassed,
S – spectrophotometer set at 240 nm,
O CH3 , HCl – a 10 µl loop injector.
N Inject the reference solution and measure the peak responses
O O of all the peaks. The relative retention times for desacetyl
diltiazem and diltiazem are about 0.65 and 1.0 respectively.
N The resolution between desacetyl diltiazem and diltiazem is
H3 C CH3 not less than 3, and the theoretical plates for the diltiazem
peak is not less than 1200. The relative standard deviation of
C22H26N2O4S,HCl Mol. Wt. 451.0 the peak response for replicate injections due to diltiazem
hydrochloride and desacetyl diltiazem is not more than 2.0 per
Diltiazem Hydrochloride is s-2,3,4,5-tetrahydro-5-(2- cent.
dimethylaminoethyl)-2-(4-methoxyphenyl)-4-
oxobenzo[b]thiazepin-3-yl acetate monohydrochloride. Inject the test solution and the reference solution and measure
the responses of all the peaks.
Diltiazem Hydrochloride contains not less than 98.5 per cent
and not more than 101.5 per cent of C22H26N2O4S,HCl, Calculate the percentage content of desacetyl diltiazem
calculated on the dried basis. hydrochloride in the substance under examination by
comparing the area of the peaks due to desacetyl diltiazem in
Description. A white, crystalline powder or small crystals. the chromatograms of the test solution and the reference
solution and from the content of desacetyl diltiazem
Identification
hydrochloride in the reference solution. Similarly, calculate
A. Determine by infrared absorption spectrophotometry (2.4.6). the percentage content of each impurity peak other than the
Compare the spectrum with that obtained with diltiazem peaks due to diltiazem and desacetyl diltiazem with that of the
hydrochloride RS. peak due to desacetyl diltiazem in the chromatogram obtained
423
DILTIAZEM TABLETS IP 2007
with the test solution and from the content of desacetyl Dissolution (2.5.2).
diltiazem hydrochloride in the reference solution. Desacetyl Apparatus. No 1
diltiazem hydrochloride content is not more than 0.5 per cent
Medium. 900 ml of freshly distilled water
w/v, the total impurities including desacetyl diltiazem
Speed and time. 100 rpm and 30 minutes and 3 hours
hydrochloride content is not more than 1 per cent w/v with no
individual impurity more than 0.5 per cent w/v. Withdraw a suitable volume of the medium after 30 minutes
Heavy metals (2.3.13). 1.0 g complies with the limit test for and 3 hours. Filter promptly through a membrane filter disc
heavy metals. Method A (20 ppm). with an average pore diameter not greater than 1.0 µm, rejecting
the first few ml of the filtrate. Dilute a suitable volume of the
Loss on drying (2.4.19). Not more than 0.5 per cent, determined filtrate with the same solvent and measure the absorbance of
on 1.0 g by drying in an oven at 105° for 2 hours. the resulting solution at the maximum at about 240 nm (2.4.7).
Assay. Determine by liquid chromatography (2.4.14) as given Calculate the content of C22H26N2O4S,HCl from the absorbance
under the test for Related substances using the following obtained from a solution of known concentration of diltiazem
solutions. hydrochloride RS.
Test solution. Dissolve 0.12 g of the substance under Use the following acceptance criteria for the 30-minute time
examination in 100 ml of methanol. interval. At S1: no unit is more than D; at S2, the average value
is equal to or less than D, and no unit is greater than D + 10 per
Reference solution (a). A solution containing 0.0012 per cent cent; at S3. the average value is equal to or less than D, not
w/v each of diltiazem hydrochloride RS and desacetyl more than 2 units are more than D + 10 per cent and no unit is
diltiazem hydrochloride RS in methanol. more than D + 25 per cent. Use the acceptance criteria in
Reference solution (b). Dissolve 60 mg of diltiazem Acceptance Table 1(2.5.2) for the 3-hour time interval.
hydrochloride RS in 50 ml of methanol. D. Not less than 60 per cent of the stated amount of
Inject reference solution (a) and check the system suitability C22H26N2O4S,HCl is dissolved in 30 minutes and not less than
parameters like the relative retention times, the resolution and 80 per cent is dissolved in 3 hours.
the column efficiency in terms of theoretical plates. Assay. Determine by liquid chromatography (2.4.14).
Inject reference solution (b). The test is not valid unless the Test solution. Weigh and powder 20 tablets. Weigh accurately
relative standard deviation for replicate injections is not more a quantity of the powder containing about 0.6 g of Diltiazem
than 2.0 per cent. Hydrochloride, add 200 ml of methanol, mix with the aid of
Inject alternately the test solution and reference solution (b). ultrasound for 1 hour, cool and dilute to 500.0 ml with methanol;
centrifuge 25 ml at 3500 rpm for 15 minutes and use the clear,
Calculate the content of C22H26N2O4S,HCl.
supernatant liquid.
w/v each of diltiazem hydrochloride RS and desacetyl
diltiazem hydrochloride RS in methanol.
Reference solution (b). Dissolve 60 mg of diltiazem
Diltiazem Tablets hydrochloride RS in 50 ml of methanol.
Diltiazem Hydrochloride Tablets Chromatographic system
Diltiazem Tablets contain not less than 90.0 per cent and not – a stainless steel column 30 cm x 3.9 mm, packed with
more than 110.0 per cent of the stated amount of diltiazem octadecylsilyl silica gel (5 µm),
hydrochloride, C22H26N2O4S, HCl. The tablets may be coated. – mobile phase: a mixture of 50 volumes of a buffer solution
containing 0.116 per cent w/v of d-10-
Identification camphorsulphonic acid in 0.1 M sodium acetate, with
the pH adjusted to 6.2 by the addition of 0.1 M sodium
In the Assay, the principal peak in the chromatogram obtained hydroxide, 25 volumes of acetonitrile and 25 volumes
with the test solution corresponds to the peak due to diltiazem of methanol, filtered and degassed,
hydrochloride in the chromatogram obtained with the reference – flow rate. 1.6 ml per minute,
solution. – spectrophotometer set at 240 nm,
Tests
Inject reference solution (a) and measure the peak responses
Other tests. Comply with the tests stated under Tablets. of all the peaks. The relative retention times for desacetyl
424
IP 2007 DIMERCAPROL INJECTION
diltiazem and diltiazem are about 0.65 and 1.0 respectively. Weight per ml (2.4.29). 1.238 g to 1.240 g.
The resolution between desacetyl diltiazem and diltiazem is Iron (2.3.14). Ignite 2.0 g with 1 g of anhydrous sodium
not less than 3, and the theoretical plates for the diltiazem carbonate, cool, dissolve the residue in 15 ml of dilute
peak is not less than 1200. hydrochloric acid and dilute to 45 ml with water; the resulting
Inject reference solution (b).The test is not valid unless the solution complies with the limit test for iron (20 ppm).
Halides. To 2.0 g add 25 ml of 0.5 M ethanolic potassium
than 2.0 per cent.
hydroxide and heat under a reflux condenser for 2 hours.
Inject alternately the test solution and reference solution (b). Remove the ethanol by evaporation in a current of warm air,
add 20 ml of water and cool. Add a mixture of 10 ml of strong
Calculate the content of C22H26N2O4S,HCl in the tablets..
hydrogen peroxide solution and 40 ml of water. Boil gently
Storage. Store protected from light. for 10 minutes; cool and filter rapidly. Add 10 ml of dilute
nitric acid and 5 ml of 0.1 M silver nitrate and titrate with
0.1 M ammonium thiocyanate using ferric ammonium
sulphate solution as indicator. Repeat the operation without
Dimercaprol the substance under examination. The difference in the
volumes of 0.1 M ammonium thiocyanate used in the two
B.A.L.
titrations is not more than 1.0 ml.
HO OH Assay. Weigh accurately about 0.1 g, dissolve in 40 ml of
SH methanol and add 20 ml of 0.1 M hydrochloric acid and
50.0 ml of 0.05 M iodine. Allow to stand for 10 minutes and
C3H8OS2 Mol. Wt. 124.2 titrate with 0.1 M sodium thiosulphate. Repeat the operation
Dimercaprol is (RS)-2,3-dimercaptopropanol. without the substance under examination. The difference
between the titrations represents the amount of iodine required.
Dimercaprol contains not less than 98.5 per cent w/w and not
more than 101.5 per cent w/w of C3H8OS2. 1 ml of 0.05 M iodine is equivalent to 0.00621 g of C3H8OS2.
Description. A clear, colourless or slightly yellow liquid; odour, Storage. Store protected from light in well-filled containers in
strong, characteristic and alliaceous. a refrigerator (2° to 8°).
Identification
A. Dissolve 0.1 ml in 4 ml of water and to 2 ml of the solution Dimercaprol Injection
add lead acetate solution; a yellow precipitate is obtained.
B. To 2 ml of the solution prepared for test A add 1 ml of B.A.L. Injection
0.05 M iodine; the colour of iodine is immediately discharged. Dimercaprol injection is a sterile solution of Dimercaprol in a
C. In a ground-glass-stoppered tube suspend 0.6 g of sodium mixture of Benzyl Benzoate and Arachis Oil.
bismuthate, previously heated to 200° for 2 hours, in a mixture Dimercaprol Injection contains not less than 90.0 per cent and
of 6 ml of water and 2.8 ml of a 10 per cent w/w solution of more than 110.0 per cent of the stated amount of dimercaprol,
phosphoric acid. Add 0.2 ml of the substance under C3H8OS2.
examination, mix and allow to stand for 10 minutes shaking
Description. A clear, yellow, viscous solution, having a
frequently. To 1 ml of the supernatant liquid add 5 ml of a
pungent, disagreeable odour.
0.4 per cent w/v solution of chromotropic acid sodium salt in
sulphuric acid, mix and heat for 15 minutes in a water-bath; a Tests
violet-red colour is produced.
Acidity. Shake with an equal volume of water for 2 minutes
Tests and set aside for separation; pH of the aqueous layer after
filtration through a neutral filter is 4.5 to 6.5 (2.4.24).
Appearance of solution. The substance under examination is
clear (2.4.1), and not more intensely coloured than reference Refractive index (2.4.27). 1.481 to 1.486, determined at 20°.
solution BS6 or BYS6 (2.4.1). Weight per ml (2.4.29). About 0.95 g.
pH (2.4.24). 5.0 to 6.5, determined in a saturated solution. Other tests. Complies with the tests stated under Parenteral
Refractive index (2.4.27). 1.568 to 1.574, determined at 20°. Preparations (Injections).
425
ACTIVATED DIMETHICONE IP 2007
Assay. Weigh accurately about 1.0 g, add 20 ml of 0.1 M Shake the second tube for about 10 seconds and heat on a
hydrochloric acid and titrate with 0.05 M iodine. water-bath for 5 minutes; the solution is violet.
1 ml of 0.05 M iodine is equivalent to 0.00621 g of C3H8OS2. C. To 50 mg in a platinum crucible add 0.15 ml of sulphuric
acid and ignite until a white residue is obtained; the residue
Determine the weight per ml of the injection (2.4.29), and
gives the reaction of silicates (2.3.1).
calculate the percentage w/v of C3H8OS2.
Storage. Store protected from light. Tests
Labelling. The label states (1) the nature of the solvent; (2)
Acidity. To 2.0 g add 25 ml of a mixture of equal volumes of
that the injection is meant for intramuscular use only.
ethanol and ether previously neutralised to 0.2 ml of
bromothymol blue solution and shake; not more than 0.15 ml
of 0.01 M sodium hydroxide is required to change the colour
Activated Dimethicone of the solution to blue.
Simethicone; Activated Polydimethylsiloxane Heavy metals (2.3.13). 2.0 g complies with the limit test for
CH3 CH3 Defoaming activity. Weigh accurately about 0.2 g, transfer to

a 100-ml bottle, add 50 ml of 2-methylpropanol and shake
H3C Si O Si CH3 + SiO2
vigorously, warming, if necessary, to effect solution. Add
CH3 CH3 dropwise, 0.5 ml of this solution to a clean, unused, cylindrical
n
250-ml glass jar, fitted with a 50-mm cap, containing 100 ml of
(CH3)3Si[OSi(CH3)2]CH3+SiO2 a 1.0 per cent w/v solution of octoxinol. Cap the jar and clamp
it in an upright position in a wrist-action shaker capable of
Activated Dimethicone is a mixture of fully methylated linear moving the jar through a radius of 13.3 ± 0.4 cm (measured
siloxane polymers containing repeating -(CH3)2SiO- units from the centre of the shaft to the centre of the jar) and an arc
stabilised with trimethylsiloxy, (CH3)3SiO-, end-blocking units of 10 degrees at a frequency of 300 ± 30 strokes per minute.
and finely divided silicon dioxide. Shake for 10 seconds and record the time required in seconds
Activated Dimethicone contains not less than 90.0 per cent for the foam to collapse. The time for foam collapse is
and not more than 99.0 per cent of polydimethylsiloxane, determined at the instant the first portion of foam-free liquid
[-(CH3)2SiO-]n and not less than 4.0 per cent and not more than surface appears, measured from the end of the shaking period.
7.0 per cent of silicon dioxide, SiO2. The defoaming activity time is not more than 15 seconds.
Description. A translucent, grey viscous liquid; almost Assay. For polydimethylsiloxane - Weigh accurately about
odourless. 50 mg, transfer to a narrow-mouthed glass bottle and add
25 ml of carbon tetrachloride. Swirl to disperse, add 50 ml of
Identification dilute hydrochloric acid, close the bottle securely with a cap
having an inert liner and shake for exactly 5 minutes. Transfer
A. To 50 mg add 25 ml of carbon tetrachloride and swirl to the mixture to a 125-ml separating funnel and remove about
disperse. Add 50 ml of dilute hydrochloric acid and shake for 5 ml of the lower layer to a stoppered test-tube containing
5 minutes. Transfer to a separating funnel and remove about 0.5 g of anhydrous sodium sulphate. Close the test-tube,
5 ml of the lower layer to a stoppered tube containing 0.5 g of agitate vigorously and centrifuge the mixture until a clear
anhydrous sodium sulphate. Shake vigorously and centrifuge supernatant liquid is obtained. Prepare a blank by mixing 10 ml
the mixture until a clear supernatant liquid is obtained. The of carbon tetrachloride with 0.5 g of anhydrous sodium
resulting liquid complies with the following test. sulphate and centrifuging to obtain a clear supernatant liquid.
Determine by infrared absorption spectrophotometry (2.4.6). Determine the absorbance of a 0.5-mm layer of the solution at
Compare the spectrum with that obtained with dimethicone the maximum at about 7.9 µm in a suitable infra-red
RS. Ignore the region of the spectrum from 850 to 750 cm-1 spectrophotometer (2.4.6), using the blank to set the
since slight differences may be observed depending on the instrument. Calculate the content of [-(CH3)2SiO-]n from the
degree of polymerisation. absorbance obtained by repeating the Assay on a 0.2 per cent
w/v solution of dimethicone RS in place of the substance
B. Heat 0.5 g in a test-tube over a small flame until white fumes
under examination.
are evolved. Invert the test-tube over a second tube containing
1 ml of a 0.1 per cent w/v solution of chromotropic acid sodium For silicon dioxide - Mix thoroughly and weigh accurately
salt in sulphuric acid so that the fumes reach the solution. about 1.0 g; transfer to a tared, sintered-glass filtering crucible
426
IP 2007 DIPHENHYDRAMINE CAPSULES
(porosity No. 4) and pass through the filter, with suction, Tests
200 ml of carbon tetrachloride, added with stirring in small
portions, followed by similar washing of the material on the Appearance of solution. A 5.0 per cent w/v solution in carbon
filter with 200 ml of n-hexane, and discard the filtrates. Place dioxide-free water, and a 5-fold dilution thereof, are both clear
the filtering crucible in a muffle furnace at room temperature, (2.4.1). The 5.0 per cent solution is not more intensely coloured
raise the temperature of the furnace to 550°. Heat at 550° ± 25° than reference solution BYS6 (2.4.1).
for 2 hours. Cool the filtering crucible with its contents in a pH (2.4.24). 4.0 to 6.0, determined in a 5.0 per cent w/v solution.
desiccator, weigh and calculate the content of silicon dioxide,
SiO2, in the sample taken.
(2.4.17), coating the plate with silica gel H.
20 volumes of methanol and 1 volume of diethylamine.
Diphenhydramine Hydrochloride Test solution. Dissolve 0.2 g in 10 ml of methanol immediately
before use.
CH3 Reference solution. Dilute 1 ml of the test solution to 100 ml
N with methanol immediately before use.
O CH3
, HCl phase to rise 10 cm. Dry the plate in air for 5 minutes, spray
with sulphuric acid and heat at 120° for 10 minutes until spots
appear. Any secondary spot in the chromatogram obtained
C17H21NO,HCl Mol. Wt. 291.8 with the test solution is not more intense than the spot in the
Diphenhydramine Hydrochloride is 2-benzhydryloxy-
ethyldimethylamine hydrochloride. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Diphenhydramine Hydrochloride contains not less than Loss on drying (2.4.19). Not more than 0.5 per cent, determined
99.0 per cent and not more than 101.0 per cent of C17H21NO,HCl, on 1.0 g by drying in an oven at 105°.
calculated on the dried basis. Assay. Weigh accurately about 0.5 g, dissolve in 20 ml of
Description. A white or almost white, crystalline powder; anhydrous glacial acetic acid and add 10 ml of mercuric
odourless or almost odourless. acetate solution. Titrate with 0.1 M perchloric acid, using
C17H21NO,HCl.
B and C may be omitted if tests A and D are carried out. Storage. Store protected from light.
diphenhydramine hydrochloride RS or with the reference
spectrum of diphenhydramine hydrochloride. Examine the Diphenhydramine Capsules
substances as discs prepared using potassium chloride IR. Diphenhydramine Hydrochloride Capsules
B. When examined in the range 230 nm to 360 nm (2.4.7), a Diphenhydramine Capsules contain not less than 93.0 per
0.025 per cent w/v solution in ethanol (95 per cent) shows cent and not more than 107.0 per cent of the stated amount of
absorption maxima at about 253 nm, 258 nm and 264 nm; diphenhydramine hydrochloride, C17H21NO,HCl.
absorbance at about 253 nm, about 0.31, at about 258 nm,
about 0.38 and at about 264 nm, about 0.3. Identification
C. To 0.05 ml of a 5 per cent w/v solution add 2 ml of sulphuric
Extract a quantity of the contents of the capsules containing
acid; an intense yellow colour develops which changes to
0.1 g of Diphenhydramine Hydrochloride with two quantities,
red on the addition of 0.5 ml of nitric acid. Add 15 ml of water,
each of 15 ml, of chloroform. Evaporate the combined extracts
cool, add 5 ml of chloroform and shake; an intense violet
to dryness on a water-bath and dry the residue at 80° for
colour develops in the chloroform layer.
1 hour. The residue melts at about 168° (2.4.21), and complies
D. Gives the reactions of chlorides (2.3.1). with the following tests.
427
DIPHENOXYLATE HYDROCHLORIDE IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). 1 ml of 0.1 M hydrochloric acid is equivalent to 0.02918 g of
Compare the spectrum with that obtained with C17H21NO,HCl.
diphenhydramine hydrochloride RS or with the reference Storage. Store protected from moisture.
spectrum of diphenhydramine hydrochloride. Examine the
substances as discs prepared using potassium chloride IR.
0.025 per cent w/v solution in ethanol (95 per cent) shows Diphenoxylate Hydrochloride
absorption maxima at about 253 nm, 258 nm and 264 nm;
absorbance at about 253 nm, about 0.31, at about 258 nm,
about 0.38 and at about 264 nm, about 0.3.
C. To 0.05 ml of a 5 per cent w/v solution add 2 ml of sulphuric O CH3
acid; an intense yellow colour develops which changes to , HCl
N
red on the addition of 0.5 ml of nitric acid. Add 15 ml of water, O
cool, add 5 ml of chloroform and shake; an intense violet CN
colour develops in the chloroform layer.
Tests
C30H32N2O2,HCl Mol. Wt. 489.1
(2.4.17), coating the plate with silica gel H. Diphenoxylate Hydrochloride is ethyl 1-(3-cyano-3,3-
diphenylpropyl)-4-phenylpiperidine-4-carboxylate
hydrochloride.
20 volumes of methanol and 1 volume of diethylamine.
Test solution. Shake a quantity of the contents of the capsules Diphenoxylate Hydrochloride contains not less than 98.0 per
containing 100 mg of Diphenhydramine Hydrochloride with cent and not more than 102.0 per cent of C30H32N2O2,HCl,
three quantities, each of 10 ml, of chloroform, filter and calculated on the dried basis.
evaporate the combined filtrate almost to dryness; dissolve Description. A white or almost white, crystalline powder;
the residue in 5 ml of chloroform. odourless or almost odourless.
with chloroform. Identification
Apply to the plate 5 µl of each solution. Allow the mobile A. Determine by infrared absorption spectrophotometry (2.4.6).
phase to rise 10 cm. Dry the plate in air for 5 minutes, spray Compare the spectrum with that obtained with diphenoxylate
with sulphuric acid and heat at 120° for 10 minutes until spots hydrochloride RS or with the reference spectrum of
appear. Any secondary spot in the chromatogram obtained diphenoxylate hydrochloride.
with the test solution is not more intense than the spot in the B. Dissolve about 30 mg in 5 ml of methanol, add 0.25 ml of
chromatogram obtained with the reference solution. nitric acid and 0.4 ml of silver nitrate solution. Shake and
Other tests. Comply with the tests stated under Capsules. allow to stand; a curdled precipitate is formed. Centrifuge and
Assay. Weigh accurately a quantity of the mixed contents of rinse the precipitate with three quantities, each of 2 ml, of
20 capsules and transfer to a 100-ml volumetric flask, add methanol. Carry out this operation rapidly in subdued light.
sufficient water to produce 100.0 ml, shake well to dissolve Suspend the precipitate in 2 ml of water and add 1.5 ml of
and filter. To an accurately measured volume of the filtrate 10 M ammonia; the precipitate dissolves easily.
containing 0.3 g of Diphenhydramine Hydrochloride add 5 g C. Gives reaction A of chlorides (2.3.1).
of sodium chloride and 5 ml of sodium hydroxide solution
and extract with successive quantities, each of 20 ml, of ether Tests
until complete extraction is effected. Wash the combined
extracts with two quantities, each of 5 ml, of water, extract the
dichloromethane is clear (2.4.1), and not more intensely
combined washings with two quantities, each of 10 ml, of
ether, add the ether to the combined ether extracts and
evaporate to about 10 ml. Add 25.0 ml of 0.1 M hydrochloric Related substances. Determine by thin-layer chromatography
acid, warm gently to complete the removal of ether, cool and (2.4.17), coating the plate with a suitable octadecylsilyl silica
titrate the excess of acid with 0.1 M sodium hydroxide using gel (5 µm) containing a fluorescent indicator with a maximum
methyl red solution as indicator. intensity at 254 nm.
428
IP 2007 DISODIUM EDETATE INJECTION
Mobile phase. A mixture of 60 volumes of dioxan, 30 volumes Disodium Edetate is disodium ethylenediaminetetraacetate
of a 5.9 per cent w/v solution of sodium chloride and 10 volumes dihydrate.
of methanol. Disodium Edetate contains not less than 98.5 per cent and not
Test solution. Dissolve 1 g of the substance under examination more than 101.0 per cent of C10H14N2Na2O8,2H2O.
in a mixture of 1 volume of methanol and 2 volumes of dichloro-
Description. A white, crystalline powder; odourless.
methane and dilute to 20 ml with the same solvent mixture.
Reference solution (a). Dilute 0.5 ml of the test solution to Identification
100 ml with the same solvent mixture.
Reference solution (b). Dissolve 0.5 g of the substance under Compare the spectrum with that obtained with disodium
examination in 25 ml of a 1.5 per cent w/v solution of potassium edetate RS.
hydroxide in methanol and add 1 ml of water. Heat on a water-
bath under a reflux condenser for 4 hours. Cool and add 25 ml B. Dissolve 2 g in 25 ml of water, add 6 ml of lead nitrate
of 0.5 M hydrochloric acid and shake with 100 ml of solution, shake and add 3 ml of potassium iodide solution;
dichloromethane. Evaporate the organic layer to dryness on no yellow precipitate is produced. Make alkaline to red litmus
a water-bath. Dissolve the residue in 10 ml of a mixture of paper with 2 M ammonia and add 5 ml of ammonium oxalate
1 volume of methanol and 2 volumes of dichloromethane, solution; no precipitate is produced.
add 10 ml of test solution and dilute to 25 ml with a mixture of C. Dissolve 0.5 g in 10 ml of water, add 0.5 ml of a 10 per cent
1 volume of methanol and 2 volumes of dichloromethane. w/v solution of calcium chloride, make alkaline to red litmus
Apply to the plate 1 µl of each solution. Allow the mobile paper with 2 M ammonia and add 3 ml of ammonium oxalate
phase to rise 7 cm in an unsaturated tank. Dry the plate in an solution; no precipitate is produced.
oven at 160° for 15 minutes and place the plate while hot in a D. Gives the reactions of sodium salts (2.3.1).
closed tank containing 20 ml of fuming nitric acid for 30
minutes. Remove the plate and heat it again at 160° for Tests
15 minutes. Allow to cool and examine immediately in ultraviolet
light at 254 nm. Any secondary spot in the chromatogram Appearance of solution. A 5.0 per cent w/v solution in carbon
obtained with the test solution is not more intense than the dioxide-free water is clear (2.4.1), and colourless (2.4.1).
spot in the chromatogram obtained with reference solution pH (2.4.24). 4.0 to 5.5, determined in a 5.0 per cent w/v solution.
(a). The test is not valid unless the chromatogram obtained
with reference solution (b) shows two clearly separated Heavy Metals (2.3.13). 1.0 g complies with the limit test for
principal spots. heavy metals, Method A (20 ppm).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Iron (2.3.14). 20 ml of a 2.5 per cent w/v solution complies with
the limit test for iron (80 ppm). Add 0.25 g of calcium chloride
to each solution before adding mercaptoacetic acid.
Assay. Weigh accurately about 0.5 g, dissolve in sufficient
water to produce 300 ml and add 2 g of hexamine and 2 ml of
ethanol (95 per cent) and add 5 ml of 0.01 M hydrochloric
2 M hydrochloric acid. Titrate with 0.1 M lead nitrate using
acid. Titrate with 0.1 M ethanolic sodium hydroxide,
about 50 mg of xylenol orange triturate as indicator.
determining the end-point potentiometrically (2.4.25). Read
the volume added between the two points of inflection. 1 ml of 0.1 M lead nitrate is equivalent to 0.03722 g of
1 ml of 0.1 M ethanolic sodium hydroxide is equivalent to C10H14N2Na2O8,2H2O.
0.04891 g of C30H32N2O2,HCl.
Disodium Edetate Injection
Disodium Edetate Disodium Edetate Injection is a sterile solution of Disodium
Edetate in Water for Injections, containing varying amounts
of the disodium and trisodium salts as a result of pH
N N adjustment.
NaOOC COONa , 2H2O
Disodium Edetate Injection contains not less than 90.0 per
NaOOC COONa
C10H14N2Na2O8,2H2O Mol. Wt. 372.2 disodium edetate, C10H14N2Na2O8.
429
DISULFIRAM IP 2007
Identification Description. A white or almost white, crystalline powder.
To a volume containing about 3 g of Disodium Edetate add Identification

3 M hydrochloric acid to adjust the pH to 5.0 and evaporate
to dryness on a steam-bath to dryness. The residue so obtained Test A may be omitted if tests B and C are carried out. Tests B
complies with the following tests. and C may be omitted if test A is carried out.
A. Determine by infrared absorption spectrophotometry (2.4.6). A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with disodium Compare the spectrum with that obtained with disulfiram RS
edetate RS. or with the reference spectrum of disulfiram.
B. Dissolve 2 g in 25 ml of water, add 6 ml of lead nitrate B. In the test for Related substances, the principal spot in the
solution, shake and add 3 ml of potassium iodide solution; chromatogram obtained with test solution (b) corresponds to
no yellow precipitate is produced. Make alkaline to red litmus that in the chromatogram obtained with reference solution
paper with 2 M ammonia and add 5 ml of ammonium oxalate (b).
solution; no precipitate is produced. C. Dissolve about 10 mg in 10 ml of methanol and add 2 ml of
C. Dissolve 0.5 g in 10 ml of water, add 0.5 ml of a 10 per cent a 0.05 per cent w/v solution of cupric chloride in methanol; a
w/v solution of calcium chloride, make alkaline to red litmus yellow colour is produced which changes to greenish-yellow.
paper with 2 M ammonia and add 3 ml of ammonium oxalate
solution; no precipitate is produced. Tests
Tests Diethyldithiocarbamate. Dissolve 0.2 g in 10 ml of ether, add

5 ml of phosphate buffer pH 8.0, shake vigorously, reject the
pH (2.4.24). 6.5 to 7.5. ether layer and wash the aqueous layer with 10 ml of ether. To
Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit the aqueous layer add 0.2 ml of a 0.4 per cent w/v solution of
per mg of disodium edetate. cupric sulphate and 5 ml of carbon tetrachloride and shake
well. Any yellow colour in the carbon tetrachloride layer is not
Other tests. Complies with the tests stated under Parenteral more intense than that of a standard prepared at the same time
Preparations (Injections). and in the same manner using 0.2 ml of a freshly prepared
Assay. Dilute an accurately measured volume containing about 0.015 per cent w/v solution of sodium diethyldithiocarbamate
0.6 g of Disodium Edetate with water to produce 100 ml, mix in place of the substance under examination (150 ppm).
and add 2 g of hexamine and 2 ml of 2 M hydrochloric acid. Related substances. Determine by thin-layer chromatography
Titrate with 0.1 M lead nitrate using about 50 mg of xylenol (2.4.17), coating the plate with silica gel GF254.
orange triturate as indicator.
Mobile phase. A mixture of 70 volumes of n-hexane and
1 ml of 0.1 M lead nitrate is equivalent to 0.03722 g of 30 volumes of butyl acetate.
C10H14N2Na2O8,2H2O.
Storage. Store in single dose containers. examination in 10 ml of ethyl acetate.
Labelling. The label states the strength in terms of anhydrous Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with
disodium edetate contained in a suitable dose-volume. ethyl acetate.
Disulfiram with ethyl acetate.
H3C disulfiram RS with ethyl acetate.
S
H3C N S Apply to the plate 10 µl of each solution. After development,
S N CH3 dry the plate in air and examine in ultraviolet light at 254 nm.
S Any secondary spot in the chromatogram obtained with test
CH3 solution (a) is not more intense than the spot in the
C10H20N2S4 Mol. Wt. 296.5 chromatogram obtained with reference solution (a).
Disulfiram is tetraethylthiuram disulphide. Heavy metals (2.3.13). 1.0 g complies with the limit test for
heavy metals, Method C (20 ppm).
Disulfiram contains not less than 98.5 per cent and not more
than 101.0 per cent of C10H20N2S4, calculated on the dried basis. Sulphated ash (2.3.18). Not more than 0.1 per cent.
430
IP 2007 DITHRANOL
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Related substances. Determine by thin-layer chromatography
on 1.0 g by drying in an oven over phosphorus pentoxide at (2.4.17), coating the plate with silica gel GF254.
50° at a pressure of 1.5 to 2.5 kPa. Mobile phase. A mixture of 70 volumes of n-hexane and
Assay. Weigh accurately about 0.45 g and dissolve in 80 ml of 30 volumes of butyl acetate.
acetone. Add 20 ml of a 2 per cent w/v solution of potassium
Test solution (a). Extract a quantity of the powdered tablets
nitrate. Titrate with 0.1 M silver nitrate determining the end-
containing 0.5 g of Disulfiram with 20 ml of ethyl acetate and
point potentiometrically (2.4.25), using a silver indicator
filter.
electrode and a silver-silver chloride double-junction electrode
saturated with potassium nitrate. Test solution (b). Dilute 1 ml of test solution (a) to 100 ml with
ethyl acetate.
C10H20N2S4. Reference solution. A 0.025 per cent w/v solution of disulfiram
RS in ethyl acetate.
Any secondary spot in the chromatogram obtained with test
Disulfiram Tablets solution (a) is not more intense than the spot in the
Disulfiram Tablets contain not less than 95.0 per cent and not chromatogram obtained with the reference solution.
more than 105.0 per cent of the stated amount of disulfiram, Other tests. Comply with the tests stated under Tablets.
C10H20N2S4.
quantity of the powder containing about 0.4 g of Disulfiram
Identification and shake with 75 ml of methanol for 30 minutes. Add sufficient
A. Extract a quantity of the powdered tablets containing 0.2 g methanol to produce 100.0 ml, mix, filter and dilute 5.0 ml of
of Disulfiram by boiling with 5 ml of carbon tetrachloride, the filtrate to 100.0 ml with methanol. To 5.0 ml add sufficient
filter and evaporate the filtrate to dryness. The residue, after of a 0.1 per cent w/v solution of cupric chloride in methanol
drying at 40° at a pressure not exceeding 0.7 kPa, complies to produce 25.0 ml, mix and allow to stand for 1 hour. Measure
with the following test. the absorbance of the resulting solution at the maximum at
about 400 nm (2.4.7), using as the blank a solution prepared
Determine by infrared absorption spectrophotometry (2.4.6). by diluting 5.0 ml of methanol to 25.0 ml with the cupric chloride
Compare the spectrum with that obtained with disulfiram RS solution. Calculate the content of C10H20N 2S 4 from the
or with the reference spectrum of disulfiram. absorbance obtained by repeating the operation using 5.0 ml
B. In the test for Related substances, the principal spot in the of 0.02 per cent w/v solution of disulfiram RS in methanol
chromatogram obtained with test solution (b) corresponds to beginning at the words “add sufficient of a 0.1 per cent w/v
that in the chromatogram obtained with the reference solution. solution of cupric chloride....”.
C. Extract a quantity of the powdered tablets containing 0.3 g Storage. Store protected from light.
of Disulfiram with ethanol (95 per cent), filter and evaporate
the filtrate to dryness. Dissolve the residue in 5 ml of ethanol
(95 per cent), add 1 ml of potassium cyanide solution; a
yellow colour is produced which changes to green and then Dithranol
darkens to bluish-green.
Anthralin; Dioxyanthranol
Tests
OH O OH
Diethyldithiocarbamate. Shake a quantity of the powdered
tablets containing 0.1 g of Disulfiram with 10 ml of chloroform
and filter. To the filtrate add 10 ml of 0.1 M sodium hydroxide,
shake, reject the chloroform layer and wash the aqueous layer
with three quantities, each of 10 ml, of chloroform. To the C14H10O3 Mol. Wt. 226.2
aqueous layer add 0.25 ml of a 0.4 per cent w/v solution of
Dithranol is 1,8-dihydroxyanthrone.
cupric sulphate and 2 ml of carbon tetrachloride, shake and
allow to separate. The lower layer is not more intensely Dithranol contains not less than 98.5 per cent and not more
coloured than reference solution BYS4 (2.4.1). than 101.0 per cent of C14H10O3, calculated on the dried basis.
431
DITHRANOL OINTMENT IP 2007
Description. A yellow or orange-yellow, microcrystalline Tests

powder; odourless or almost odourless.
Dihydroxyanthracene. Dissolve a quantity containing 0.1 g
Identification of Dithranol in 5 ml of hot benzene; a yellow or orange solution
is produced.
Dihydroxyanthraquinone. Dissolve a quantity containing
Compare the spectrum with that obtained with dithranol RS
1 mg of Dithranol in a few drops of sulphuric acid; an orange
or with the reference spectrum of dithranol.
solution with no trace of violet colour is produced.
Other tests. Complies with the tests stated under Ointments.
0.001 per cent w/v solution in chloroform shows absorption
maxima at about 255 nm, 287 nm and 354 nm; absorbances at Assay. Determine by liquid chromatography (2.4.14)
the maxima, about 0.55, 0.5 and 0.45 respectively. Test solution. Weigh accurately a quantity of the ointment
C. Melts at about 178° (2.4.21). containing about 5 mg of Dithranol, disperse in 20 ml of
dichloromethane, add 1.0 ml of glacial acetic acid, dilute to
Tests 100.0 ml with hexane and filter.
Dihydroxyanthracene. Dissolve 0.1 g in 5 ml of hot benzene; Reference solution. Add 1.0 ml of glacial acetic acid to
a clear yellow or orange solution is produced. 20.0 ml of a 0.025 per cent w/v solution of dithranol RS in
dichloromethane and add sufficient hexane to produce
Dihydroxyanthraquinone. Dissolve 1 mg in a few drops of 100.0 ml.
sulphuric acid; a clear orange solution with no trace of violet
colour is produced. Chromatographic system
Sulphated ash (2.3.18). Not more than 0.1 per cent. porous silica particles (5 µm),
Loss on drying (2.4.19). Not more than 0.5 per cent, determined – mobile phase: a mixture of 82 volumes of hexane,
on 1.0 g by drying in an oven at 105°. 5 volumes of dichloromethane and 1 volume of glacial
acetic acid,
anhydrous pyridine. Titrate with 0.1 M tetrabutylammonium
(2.4.25). Carry out a blank titration.
Inject the test solution and reference solution.
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to Calculate the content of C14H10O3 in the ointment.
0.02262 g of C14H10O3. Storage. Store protected from light.
Docusate Sodium
Dioctyl Sodium Sulphosuccinate
Dithranol Ointment
Dithranol Ointment contains Dithranol, in fine powder, in a CH3
O
suitable base. O CH3
Na H3C O
Dithranol Ointment contains not less than 90.0 per cent and
SO3 O
not more than 110.0 per cent of the stated amount of dithranol, H3C
C14H10O3.
C20H37NaO7S Mol. Wt.444.6
Identification Docusate Sodium is sodium 1,4-bis[(2-ethylhexyl)oxy]-1,4-
A. In the Assay, the principal peak in the chromatogram dioxobutane-2-sulphonate.
obtained with the test solution corresponds to the peak in the Docusate Sodium contains not less than 98.0 per cent and not
chromatogram obtained with the reference solution. more than 101.0 per cent of C20H37NaO7S, calculated on the
B. Heat a quantity containing 0.5 mg of Dithranol with 5 ml of anhydrous basis.
1 M sodium hydroxide on a water-bath with constant stirring; Description. White or almost white, waxy masses or flakes,
a pink colour is produced in the aqueous layer. hygroscopic.
432
IP 2007 DOMPERIDONE MALEATE
Identification Not more than 0.5 ml of 0.1 M silver nitrate is required to

change the colour of the indicator from yellow to orange ( 350
A. Determine by infrared absorption spectrophotometry (2.4.6). ppm).
Compare the spectrum with that obtained with docusate sodium
RS. Sodium sulphate. Not more than 2 per cent.
B. Ignite 0.75 g in the presence of dilute sulphuric acid, until Dissolve 0.25 g in 40 ml of a mixture of 20 volumes of water
an almost white residue is obtained. Cool and add 5 ml of and 80 volumes of 2-propanol. Adjust to pH between 2.5 and
water, Filter. 2 ml of the filtrate gives reaction (a) of sodium 4.0 using perchloric acid solution. Add 0.4 ml of naphtharson
salts (2.3.1). solution and 0.1 ml of 0.0125 per cent w/v solution of methylene
blue. Not more than 1.5 ml of 0.025 M barium perchlorate is
Tests required to change the colour of the indicator from yellowish-
green to yellowish-pink.
Alkalinity. Dissolve 1.0 g in 100 ml of a mixture of equal
volumes of methanol and water, previously neutralised to Heavy metals (2.3.13). Dissolve 4.0 g in 20 ml of alcohol
methyl red solution. Add 0.1 ml of methyl red solution. Not (80 per cent v/v). 12 ml of the solution complies with the limit
more than 0.2 ml of 0.1 M hydrochloric acid is required to test for heavy metals, Method B (10 ppm ).
change the colour of the indicator to red. Water (2.3.43). Not more than 3.0 per cent, determined on
Related non-ionic substances. Determine by gas 0.25 g.
Internal standard solution. Dissolve 10 mg of methyl behenate 0.5 M alcoholic potassium hydroxide and heat on a water-
in 50 ml of hexane. bath under reflux for 45 minutes. Cool, add 0.25 ml of
Test solution (a). Dissolve 0.1 g of the substance under phenolphthalein solution and titrate with 0.5 M hydrochloric
examination in 2.0 ml of the internal standard solution and acid until the red colour disappears. Carry out a blank titration.
dilute to 5.0 ml with hexane. Pass the solution, at a rate of 1 ml of 0.5 M hydrochloric acid is equivalent to 0.1112 g of
about 1.5 ml per minute, through a column 10 mm in internal C20H37NaO7S.
diameter, packed with 5 g of basic aluminium oxide and
previously washed with 25 ml of hexane. Elute with 5 ml of
hexane and discard the eluate. Elute with 20 ml of a mixture of
equal volumes of ether and hexane. Evaporate the eluate to
dryness and dissolve the residue in 2.0 ml of hexane. Domperidone Maleate
Test solution (b). Prepare as described for test solution (a)
but dissolving 0.1 g of the substance under examination in N N
5.0 ml of hexane and using a new column. O
N COOH
Reference solution. Dilute 2.0 ml of the internal standard ,
H N
solution to 5.0 ml with hexane.
O COOH
Chromatographic system N
– a glass column 2 m x 2 mm, packed with silanised Cl
H
diatomaceous earth for gas chromatography (150 µm to
180 µm) impregnated with 3 per cent m/m of C22H24ClN5O2,C4H4O4 Mol. Wt. 542.0
polymethylphenylsiloxane, Domperidone Maleate is 5-chloro-1-[1-[3-(2,3-dihydro-2-
– temperature : oxo-1H-benzimidazol-1-yl)propyl]-4-piperidinyl]-1,3-
column 230°, dihydro-2H-benzimidazol-2-one maleate.
Inlet port and detector at 280°,
– flow rate. 30 ml per minute of the nitrogen carrier gas. Domperidone Maleate contains not less than 99.0 per cent
and not more than 101.0 per cent of domperidone maleate,
Inject 1 ml of the test solution (a), (b) and the reference solution.
C22H24ClN5O2,C4H4O4, calculated on the dried basis.
There is no peak with the same retention time as the internal
standard in the chromatogram obtained with test solution (b). Description. A white or almost white powder.
The area of any impurity peak is not more than the area of the
peak due to the internal standard (0.4 per cent). Identification
Chlorides (2.3.12). Dissolve 5.0 g in 50 ml of alcohol (50 per Determine by infrared absorption spectrophotometry (2.4.6).
cent v/v) and add 0.1 ml of potassium dichromate solution. Compare the spectrum with that obtained with domperidone
433
DOMPERIDONE TABLETS IP 2007
maleate RS or with the reference spectrum of domperidone the mobile phase or adjust the time programme for the linear
maleate. If the spectra obtained show differences, dissolve gradient.
the substance under examination and the reference substance Inject dimethylformamide as a blank, the test solution and
separately in the minimum volume of 2-propanol, evaporate reference solution (b). In the chromatogram obtained with the
to dryness on a water-bath and record new spectra using the test solution, the area of any peak, other than the principal
residues. peak, is not greater than the area of the principal peak in the
Tests cent); the sum of the areas of all peaks, other than the principal
Appearance of solution. Dissolve 0.2 g in dimethylformamide peak, is not greater than twice the area of the principal peak in
and dilute to 20.0 ml with the same solvent. The solution is the chromatogram obtained with reference solution (b) (0.5
clear (2.4.1) and not more intensely coloured than reference per cent). Ignore any peak in the chromatogram obtained with
solution Y6 (2.4.1). the blank run, any peak due to maleic acid at the beginning of
the chromatogram and any peak with an area less than 0.2
Related substances. Determine by liquid chromatography times that of the principal peak in the chromatogram obtained
(2.4.14). with reference solution (b).
NOTE - Prepare the solutions immediately before use. Heavy metals (2.3.13.). 1.0 g complies with the limit test for
Test solution. Dissolve 0.1 g of the substance under heavy metals, Method B (20 ppm). Prepare the standard using
examination in dimethylformamide and dilute to 10 ml with the 2 ml of lead standard solution (10 ppm Pb).
same solvent. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Reference solution (a). Dissolve 10 mg of domperidone Loss on drying (2.4.19). Not more than 0.5 per cent, determined
maleate RS and 15 mg of droperidol RS in dimethylformamide on 1.0 g by drying in an oven at 100º to 105º.
and dilute to 100 ml with the same solvent. Assay. Dissolve 0.4 g in 50 ml of anhydrous acetic acid.
Reference solution (b). Dilute 1 ml of the test solution to 100 Titrate with 0.1 M perchloric acid using 0.2 ml of
ml with dimethylformamide. Dilute 5 ml of the solution to 20 naphtholbenzein solution as indicator, until the colour
ml with dimethylformamide. changes from orange-yellow to green.
Chromatographic system 1 ml of 0.1 M perchloric acid is equivalent to 0.0542 g of
– a stainless steel column 10 cm x 4.6 mm, packed with C26H28ClN5O6.
base-deactivated octadecylsilyl silica gel (3 µm), Storage. Store protected from light.
– mobile phase: a mixture of 3 volumes of methanol and 7
volumes of a 0.5 per cent w/v solution of ammonium
acetate,
– a linear gradient programme with the mobile phase
Domperidone Tablets
changing to methanol over 10 Domperidone Tablets contain not less than 95.0 per cent and
minutes, followed by elution with methanol for 2 not more than 105.0 per cent of the stated amount of
minutes, domperidone, C22H24ClN5O2.
– a 10 µl loop injector. Identification
Equilibrate the column for at least 30 minutes with methanol A. Determine by thin-layer chromatography (2.4.17), coating
and then equilibrate with the initial mobile phase. the plate with silica gel F254 or using a precoated plate (such
as Merck silica gel 60 F254 plates).
principal peak in the chromatogram obtained with reference Mobile phase. A mixture of 5 volumes of a solution prepared
solution (b) is at least 50 per cent of the full scale of the by dissolving 1.36 g of sodium acetate in 50 ml of water,
recorder. adjusting the pH to 4.7 with dilute acetic acid and adding
sufficient water to produce 100 ml, 18 volumes of methanol,
Inject reference solution (a). The retention times are:
23 volumes of dichloromethane and 54 volumes of ethyl
domperidone maleate, about 6.5 minutes and droperidol, about
acetate.
7 minutes. The test is not valid unless the resolution between
the peaks due to domperidone maleate and droperidol is at Test solution. Shake a quantity of the powdered tablets
least 2.0. If necessary adjust the concentration of methanol in containing 10 mg of domperidone with 10 ml of a mixture of
434
IP 2007 DOMPERIDONE TABLETS
equal volumes of dichloromethane and methanol and filter – mobile phase: A. methanol
through a glass microfibre filter (such as Whatman GF/C). B. a 0.5 per cent w/v solution of
Reference solution. A 0.127 per cent w/v solution of ammonium acetate,
domperidone maleate RS in a mixture of equal volumes of – flow rate. 1.5 ml per minute,
dichloromethane and methanol. – a linear gradient programme using the conditions given
below,
allow the plate to dry in air and examine in ultraviolet light at – a 10 µl loop injector.
254 nm. Spray the plate with potassium iodobismuthate
Time mobile phase A mobile phase B
solution and examine again. With each method of visualisation,
the principal spot in the chromatogram obtained with the test
solution corresponds to that in the chromatogram obtained 0 30 70
with the reference solution. 10 100 0
B. In the Assay, the principal peak in the chromatogram 12 100 0
obtained with the test solution corresponds to the peak in the Equilibrate the column for at least 30 minutes with methanol
chromatogram obtained with the reference solution. and equilibrate with the initial mobile phase for at least 5
minutes. Adjust the sensitivity of the system so that the height
Tests of the principal peak in the chromatogram obtained with
Dissolution (2.5.2). reference solution (a) is at least 50 per cent of the full scale of
the recorder.
Apparatus No.1
Medium. 900 ml of 0.1 M hydrochloric acid Inject a mixture of equal volumes of 0.01 M hydrochloric acid
Speed and time. 50 rpm and 45 minutes. and methanol as a blank, the test solution and reference
solutions (a) and (b). The test is not valid unless, in the
chromatogram obtained with reference solution (b), the
the absorbance of the filtrate, diluted with the dissolution
resolution between the two principal peaks is at least 2. If
medium if necessary, at 286 nm (2.4.7), using the dissolution
necessary adjust the concentration of methanol in the mobile
medium as the blank. Calculate the content of C22H24ClN5O2 in
phase or adjust the time programme for the linear gradient.
the medium from the absorbance obtained from a solution of
known concentration domperidone maleate RS. In the chromatogram obtained with the test solution the area
of any secondary peak is not greater than the area of the
(2.4.14).
solution (a) (0.25 per cent) and the sum of the areas of any
Note - Prepare the following solutions immediately before secondary peaks is not greater than twice the area of the
use. principal peak in the chromatogram obtained with the reference
Test solution. To a quantity of the powdered tablets containing solution (a) (0.5 per cent). Ignore any peak in the
50 mg of domperidone add 10 ml of a mixture of equal volumes chromatogram obtained with the blank solution and any peak
of 0.01 M hydrochloric acid and methanol, mix with the aid with an area less than 0.2 times the area of the peak in the
of ultrasound for 20 minutes and filter through a glass chromatogram obtained with reference solution (a) (0.05 per
microfibre filter (such as Whatman GF/C). cent).
Reference solution (a). Dilute 1 volume of the test solution to Other tests. Comply with the tests stated under Tablets.
200 volumes with a mixture of equal volumes of 0.01 M Assay. Determine by liquid chromatography (2.4.14) as
hydrochloric acid and methanol. Dilute 1 volume of the described under Related substances using the following
resulting solution to 2 volumes with a mixture of equal volumes solutions.
of 0.01 M hydrochloric acid and methanol.
Test solution. Add sufficient methanol to 10 whole tablets to
Reference solution (b). A solution containing 0.01 per cent w/ produce a solution containing 0.02 per cent w/v of
v of domperidone maleate RS and 0.015 per cent w/v of domperidone, mix with the aid of ultrasound for 20 minutes
droperidol RS in a mixture of equal volumes of 0.01 M and filter through a glass microfibre filter (such as Whatman
hydrochloric acid and methanol. GF/C). To 50.0 ml of the filtrate add 1 ml of 0.1 M hydrochloric
Chromatographic system acid and sufficient water to produce 100.0 ml.
– a stainless steel column 10 cm x 4.6 mm, packed with Reference solution. A solution containing 0.0127 per cent w/v
base-deactivated, end-capped octadecylsilyl silica gel of domperidone maleate RS in a mixture of equal volumes of
(3 µm) (such as Hypersil BDS). 0.002 M hydrochloric acid and methanol.
435
DONEPEZIL HYDROCHLORIDE IP 2007
Calculate the content of C22H24ClN5O2 in the tablets. – spectrophotometer set at 230 nm,
Storage. Store protected from moisture. – a 20 µl loop injector.
equivalent amount of domperidone.
Donepezil Hydrochloride secondary peak is not more than 0.5 times the area of the peak
O N per cent) and the sum of areas of all the secondary peaks is
H3CO not more than twice the area of the peak in the chromatogram
, HCl obtained with the reference solution (b) (2.0 per cent).
H3CO Heavy metals (2.3.13). The residue obtained from residue on
ignition complies with limit test for heavy metals, Method B
C24H29NO3,HCl Mol. Wt. 415.5 (20 ppm).
Donepezil Hydrochloride is (±)-2-[(1-Benzyl-4-piperidyl) Sulphated ash (2.3.19). Not more than 0.2 per cent.
methyl]-5,6-dimethoxy-1-indanone hydrochloride. Water (2.3.43). Not more than 7.0 per cent, determined on
Donepezil Hydrochloride contains not less than 98.0 per cent 1.0 g.
and not more than 102.0 per cent of C24H29NO3,HCl, calculated Assay. Weigh accurately about 0.4 g, dissolve in a mixture of
on the anhydrous basis. 40 ml of glacial acetic acid and 10 ml of 5 per cent of mercuric
Description. A white to off-white powder. acetate in glacial acetic acid. Titrate with 0.1 M perchloric
acid. Determine the end-point potentiometrically (2.4.25). Carry
Identification out a blank titration.
A. Determine by infrared absorption spectrophotometry (2.4.6). 1 ml of 0.1 M perchloric acid is equivalent to 0.04155 g of
Compare the spectrum with that obtained with donepezil C24H29NO3,HCl.
hydrochloride RS or with the reference spectrum of donepezil Storage. Store protected from moisture.
hydrochloride.
B. When examined in the range 200 nm to 400 nm ( 2.4.7), a
maximum at about 230 nm, 268 nm and 313 nm.
Donepezil Tablets
Tests Donepezil Hydrochloride Tablets
Related substances. Determine by liquid chromatography Donepezil Tablets contain not less than 90.0 per cent and not
(2.4.14). more than 110.0 per cent of the stated amount of donepezil
hydrochloride, C24H29NO3,HCl.
examination in 50 ml of mobile phase. Identification
Reference solution (a). A 0.1 per cent w/v solution of In the Assay, the principal peak in the chromatogram obtained
donepezil hydrochloride RS in mobile phase. with the test solution corresponds to the peak in the
Reference solution (b). Dilute 1 ml of reference solution (a) to chromatogram obtained with the reference solution.
100 ml with mobile phase.
Tests
– a stainless steel column 25 cm x 4.6 mm packed with Dissolution (2.5.2).
Apparatus No. 1
400 volumes of methanol and 1 volume of triethylamine,
adjust the pH to 3.0 with orthophosphoric acid and Speed and time. 50 rpm and 45 minutes.
filter, Withdraw a suitable volume of the medium and filter. Measure
– flow rate. 1 ml per minute, the absorbance of the filtered solution, suitably diluted with
436
IP 2007 DOTHIEPIN HYDROCHLORIDE
the medium if necessary, at the maximum at about 230 nm not more than 3 times the area of the peak in the chromatogram
(2.4.7). Calculate the content of C24H29NO3,HCl in the medium obtained with the reference solution (b) (3.0 per cent).
from the absorbance obtained from a solution of known Other tests. Comply with the tests stated under Tablets.
concentration of donepezil hydrochloride RS in the same
medium. Assay. Determine by liquid chromatography (2.4.14).
D. Not less than 75 per cent of the stated amount of Test solution. Transfer intact tablets and sonicate to disperse
C24H29NO3,HCl. the tablets completely, add mobile phase and sonicate again
for 30 minutes and cool and make up the volume with mobile
Related substances. Determine by liquid chromatography phase. Allow the excipients to settle down completely and
(2.4.14). dilute finally with mobile phase to obtain a solution of final
Solvent mixture. 25 volumes of acetonitrile and 75 volumes concentration of 0.01 per cent w/v and filter.
of water. Reference solution. A 0.01 per cent w/v solution of donepezil
Test solution. Weigh accurately a quantity of powdered tablets hydrochloride RS in mobile phase.
containing about 50 mg of Donepezil Hydrochloride, add Chromatographic system
25 ml of solvent mixture, sonicate for 15 minutes and make up – a stainless steel column 25 cm x 4.6 mm packed with
the volume to 50 ml with solvent mixture and filter. octadecylsilane bonded to porous silica (5 µm),
– mobile phase: a mixture of 60 volumes of buffer pH 2.2
donepezil hydrochloride RS in solvent mixture.
prepared by dissolving about 6.8 g of potassium
Reference solution (b). Dilute 1 ml of reference solution (a) to dihydrogen phosphate in 1000 ml of water. Add 5 ml of
100 ml with solvent mixture. triethylamine and adjust the pH to 2.2 with
Chromatographic system orthophosphoric acid, filter and 40 volumes of
– a stainless steel column 25 cm x 4.6 mm packed methanol,
octadecylsilane bonded to porous silica (5 µm), – flow rate. 1.2 ml per minute,
– mobile phase: A. a buffer solution pH 6.5 prepared by – a 20 µl loop injector.
adding 1 ml of orthophosphoric acid in Inject the reference solution. The test is not valid unless the
1000 ml of water, adjust the pH to 6.5 with triethylamine column efficiency is not less than 7000 theoretical plates. The
and filter, tailing factor is not more than 1.5. The relative standard
B. acetonitrile, deviation of replicate injections is not more than 2.0 per cent.
Inject the test solution and the reference solution
below,
– flow rate. 1.5 ml per minute, Calculate the content of C24H29NO3,HCl
– spectrophotometer set at 286 nm, Storage. Store protected from moisture, at a temperature not
– a 20 µl loop injector. exceeding 25°.
(mins.) (per cent w/v) (per cent w/v)
0 75 25 Dothiepin Hydrochloride
10 40 60 Dosulepin Hydrochloride
40 40 60
CH3
41 75 25
N
50 75 25 CH3
Inject reference solution (b). Test is not valid unless the column
,HCl
effciency is not less than 20000 theoretical plates and the
tailing factor is not more than 2.0.
S
chromatogram obtained with the test solution, the area of any C19H21NS,HCl Mol. Wt. 331.9
secondary peak is not more than 1.5 times the area of the peak Dothiepin Hydrochloride is 3-(6H-dibenzo[b,e]thiepin-11-
in the chromatogram obtained with reference solution (b) (1.5 ylidene)propyldimethylamine hydrochloride, consisting
per cent) and the sum of areas of all the secondary peaks is predominantly of the E-isomer.
437
DOTHIEPIN CAPSULES IP 2007
Dothiepin Hydrochloride contains not less than 98.0 per cent Test solution. Dissolve 0.5 g of the substance under
and not more than 101.0 per cent of C19H21NS,HCl, calculated examination in 100 ml of methanol.
on the dried basis. Reference solution. A 0.5 per cent w/v solution of dothiepin
Description. A white to faintly yellow, crystalline powder; hydrochloride RS in methanol.
Identification – a glass column 1.8 m x 3 mm, packed with acid-washed,
silanised diatomaceous support (100 to 120 mesh) coated
A. Determine by infrared absorption spectrophotometry (2.4.6). with 3 per cent w/w of cyanopropylmethyl phenyl methyl
Compare the spectrum with that obtained with dothiepin silicone fluid (such as OV-225),
hydrochloride RS or with the reference spectrum of dothiepin – temperature:
hydrochloride. – column.200°,
– inlet port. 260°,
B. Dissolve 1 mg in 5 ml of sulphuric acid; a dark red colour is
produced.
– nitrogen as carrier gas.
C. On 20 mg determine by the oxygen-flask method (2.3.34),
In the chromatogram obtained with the reference solution a
using a mixture of 15 ml of water and 1 ml of hydrogen peroxide
peak due to Z-dothiepin is present with a retention time of
solution (20 volume) as the absorbing liquid. The solution
approximately 0.83 relative to the retention time of the principal
gives the reactions of sulphates (2.3.1).
peak which is due to E-dothiepin. In the chromatogram obtained
D. Gives reaction A of chlorides (2.3.1). with the test solution the area of any peak corresponding to
Z-dothiepin is not greater than 7.5 per cent of the sum of the
Tests areas of the peaks due to Z-dothiepin and E-dothiepin.
pH (2.4.24). 3.0 to 4.5, determined in a 5.0 per cent w/v solution. Heavy metals (2.3.13). Dissolve the residue obtained in the
Related substances. Determine by thin-layer chromatography test for Sulphated ash in 0.5 ml of hydrochloric acid, evaporate
(2.4.17), coating the plate with silica gel HF254. to dryness, dissolve the residue in 2 ml of water, neutralise to
phenolphthalein solution with dilute sodium hydroxide
Mobile phase. A mixture of 90 volumes of 1,2-dichloroethane, solution and dilute to 15 ml with water, 12 ml of the resulting
10 volumes of 2-propanol and 1 volume of strong ammonia solution complies with the limit test for heavy metals, Method
solution. D (20 ppm).
Prepare the following solutions freshly before use. Sulphated ash (2.3.18). Not more than 0.1 per cent, determined
Test solution (a). Dissolve 1.0 g of the substance under on 1.5 g.
examination in 10 ml of chloroform. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Test solution (b). Dissolve 0.4 g of the substance under on 1.0 g by drying in an oven at 105°.
examination in 10 ml of chloroform. Assay. Weigh accurately about 0.6 g, dissolve in 100 ml of
Reference solution. A solution containing 0.02 per cent w/v acetone and add 15 ml mercuric acetate solution. Titrate with
each of 11-(3-dimethylamino-propylidene)-6H- 0.1 M perchloric acid, using 3 ml of a saturated solution of
dibenzo[b,e]thiepin- 5-oxide RS and 6H-dibenzo- methyl orange in acetone as indicator. Carry out a blank
[b,e]thiepin-11-one RS in chloroform. titration.
Apply to the plate 5 µl of each solution. After development, 1 ml of 0.1 M perchloric acid is equivalent to 0.03319 g of
dry the plate in air and examine in ultraviolet light at 254 nm. In C19H21NS,HCl.
the chromatogram obtained with the reference solution the Storage. Store protected from light.
spot with the lower R f value is more intense than any
corresponding spot in the chromatogram obtained with test
solution (b). In the chromatogram obtained with test solution Dothiepin Capsules
(a) any secondary spot other than any spot corresponding to
the spot with the lower Rf value in the chromatogram obtained Dothiepin Hydrochloride Capsules; Dosulepin Capsules;
with the reference solution is not more intense than the Dosulepin Hydrochloride Capsules;
proximate spot in the chromatogram obtained with the
Dothiepin Capsules contain not less than 92.5 per cent and
reference solution.
not more than 107.5 per cent of the stated amount of dothiepin
Z-Isomer. Determine by gas chromatography (2.4.13). hydrochloride, C19H21NS,HCl.
438
IP 2007 DOXEPIN HYDROCHLORIDE

Extract a quantity of the contents of the capsules containing silanised diatomaceous support (100 to 120 mesh) coated
0.1 g of Dothiepin Hydrochloride with 20 ml of ethanol, filter with 3 per cent w/w of cyanopropylmethyl phenyl methyl
and remove the ethanol from the filtrate by evaporation. The silicone fluid (such as OV-225),
residue complies with the following tests. – temperature:
A. Dissolve 1 mg in 5 ml of sulphuric acid; a dark red colour is column.200°,
produced. inlet port. 260 °,
B. On 20 mg determine by the oxygen-flask method (2.3.34),
using a mixture of 15 ml of water and 1 ml of hydrogen peroxide
solution (20 volume) as the absorbing liquid. The solution In the chromatogram obtained with the reference solution a
gives the reactions of sulphates (2.3.1). peak due to Z-dothiepin is present with a retention time of
approximately 0.83 relative to the retention time of the principal
peak which is due to E-dothiepin. In the chromatogram obtained
with the test solution the area of any peak corresponding to
Tests
Z-dothiepin is not greater than 7.5 per cent of the sum of the
Related substances. Determine by thin-layer chromatography areas of the peaks due to Z-dothiepin and E-dothiepin.
(2.4.17), coating the plate with silica gel HF254. Other tests. Comply with the tests stated under Capsules.
Mobile phase. A mixture of 90 volumes of 1,2-dichloroethane, Assay. Weigh accurately a quantity of the mixed contents of
10 volumes of 2-propanol and 1 volume of strong ammonia 20 capsules containing about 0.5 g of Dothiepin Hydrochloride
solution. and extract with 20 ml followed by four quantities, each of
Test solution (a). Extract a quantity of the contents of the 10 ml, of chloroform, filtering each extract through the same
capsules containing 0.25 g of Dothiepin Hydrochloride by filter. Evaporate the combined extracts to dryness, dissolve
shaking for 2 minutes with 5 ml of chloroform, centrifuge and the residue in 100 ml of acetone and add 15 ml of mercuric
use the supernatant liquid. acetate solution. Titrate with 0.1 M perchloric acid, using
3 ml of a saturated solution of methyl orange in acetone as
Test solution (b). Dilute 2 ml of test solution (a) to 5 ml with indicator. Carry out a blank titration.
chloroform.
Reference solution. A freshly prepared solution containing C19H21NS,HCl.
0.02 per cent w/v each of 11-(3-dimethylamino-propylidene)-
6H-dibenzo[b,e]thiepin-5-oxide RS and 6H-dibenzo- Storage. Store protected from moisture.
[b,e]thiepin-11-one RS in chloroform.
Apply to the plate 5 µl of the reference solution and 10 µl of
the test solutions. After development, dry the plate in air and
examine in ultraviolet light at 254 nm. In the chromatogram Doxepin Hydrochloride
obtained with the reference solution the spot with the lower
Rf value is more intense than any corresponding spot in the CH3
chromatogram obtained with test solution (b). In the N
chromatogram obtained with test solution (a) any secondary CH3
spot other than any spot corresponding to the spot with the
lower Rf value in the chromatogram obtained with the reference ,HCl
solution is not more intense than the proximate spot in the
O
Z-Isomer. Determine by gas chromatography (2.4.13). C19H21NO,HCl Mol. Wt. 315.8
Test solution. Use the supernatant liquid obtained by extracting Doxepin Hydrochloride is 3-(6H-dibenz[b,e]oxepin-11-
a quantity of the mixed contents of 20 capsules containing ylidene)propyldimethylamine hydrochloride. It consists of a
25 mg of Dothiepin Hydrochloride with 5 ml of methanol and mixture of Z and E isomers.
centrifuging.
Doxepin Hydrochloride contains not less than 98.0 per cent
Reference solution. A 0.5 per cent w/v solution of dothiepin and not more than 101.0 per cent of C19H21NO,HCl, calculated
hydrochloride RS in methanol. on the dried basis.
439
DOXEPIN CAPSULES IP 2007
Description. A white, crystalline powder; odour, slight and with 0.1 M perchloric acid, using 3 ml of a saturated solution
amine-like. of methyl orange in acetone as indicator. Carry out a blank
titration.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). C19H21NO,HCl.
Compare the spectrum with that obtained with doxepin Storage. Store protected from light.
hydrochloride RS or with the reference spectrum of doxepin
hydrochloride.
0.004 per cent w/v solution in 0.01 M methanolic hydrochloric
Doxepin Capsules
acid shows an absorption maximum only at about 297 nm; Doxepin Hydrochloride Capsules
absorbance at about 297 nm, about 0.60.
Doxepin Capsules contain not less than 90.0 per cent and not
C. Dissolve 5 mg in 2 ml of nitric acid; a red colour is produced. more than 110.0 per cent of the stated amount of doxepin,
D. Gives reaction A of chlorides (2.3.1). C19H21NO.
Tests Identification
Z-isomer. 13.0 per cent to 18.5 per cent, determined by the Wash a quantity of the contents of the capsules containing
following method. 0.1 g of doxepin with 3 quantities, each of 5 ml, of light
petroleum (40° to 60°). Dry the residue in air and extract with
3 quantities, each of 10 ml, of chloroform, evaporate the
Test solution. Dissolve 0.5 g of the substance under combined extracts to dryness and dry the residue at 105°. The
examination in 100 ml of methanol. dried residue complies with the following tests.
Reference solution. A 0.5 per cent w/v solution of doxepin A. Determine by infrared absorption spectrophotometry (2.4.6).
hydrochloride RS in methanol. Compare the spectrum with that obtained with doxepin
Chromatographic system hydrochloride RS or with the reference spectrum of doxepin
– a glass column 1.5 m x 4 mm, packed with acid-washed, hydrochloride.
silanised diatomaceous support (100 to 120 mesh) coated B. When examined in the range 230 nm to 360 nm (2.4.7), a
with 3 per cent w/w of cyanopropylmethyl phenyl methyl 0.004 per cent w/v solution in 0.01 M methanolic hydrochloric
silicone fluid (such as OV-225), acid shows an absorption maximum only at about 297 nm;
– temperature: absorbance at about 297 nm, about 0.60.
column.200°,
C. Dissolve 5 mg in 2 ml of nitric acid; a red colour is produced.
inlet port. 260 °,
– flame ionisation detector, D. Gives reaction A of chlorides (2.3.1).
Tests
In the chromatogram obtained with the reference solution a
peak due to Z-doxepin immediately precedes and is adequately Z-isomer. 13.0 per cent to 18.5 per cent, determined by the
separated from the principal peak which is due to E-doxepin. following method.
Measure the areas or heights of the peaks due to Z-doxepin
and E-doxepin in the chromatograms obtained with the test
solution and the reference solution. Calculate the content of Test solution. Use the supernatant liquid obtained by extracting
the Z-isomer in the substance under examination. a quantity of the mixed contents of 20 capsules containing
25 mg of doxepin with 5 ml of methanol and centrifuging.
heavy metals, Method B (20 ppm). Reference solution. A 0.5 per cent w/v solution of doxepin
hydrochloride RS in methanol.
Assay. Weigh accurately about 0.6 g, dissolve in 100 ml of with 3 per cent w/w of cyanopropylmethyl phenyl methyl
acetone and add 15 ml of mercuric acetate solution. Titrate silicone fluid (such as OV-225),
440
IP 2007 DOXORUBICIN HYDROCHLORIDE
– temperature: Description. An orange-red, crystalline powder; hygroscopic.

column.200°, CAUTION - Doxorubicin Hydrochloride is poisonous. It must
inlet port. 260 °, be handled with care avoiding contact with skin and
– flame ionisation detector, inhalation of airborne particles.
In the chromatogram obtained with the reference solution a Identification
peak due to Z-doxepin immediately precedes and is adequately
separated from the principal peak which is due to E-doxepin. Test A may be omitted if tests B, C and D are carried out. Tests
Measure the areas or heights of the peaks due to Z-doxepin B and C may be omitted if tests A and D are carried out.
and E-doxepin in the chromatograms obtained with the test A. Determine by infrared absorption spectrophotometry (2.4.6).
solution and the reference solution. Calculate the content of Compare the spectrum with that obtained with doxorubicin
the Z-isomer in the capsules. hydrochloride RS.
Other tests. Comply with the tests stated under Capsules. B. When examined in the range 220 nm to 550 nm (2.4.7), a
Assay. Weigh accurately a quantity of the mixed contents of 0.001 per cent w/v solution in ethanol (95 per cent) exhibits
20 capsules containing about 30 mg of doxepin, add 50 ml of maxima at about 234 nm, 252 nm, 288 nm, 475 nm, 495 nm and
0.1 M methanolic hydrochloric acid, shake for 30 minutes 530 nm.
and add sufficient 0.01 M methanolic hydrochloric acid to C. In the test for Related substances, the principal peak in the
produce 100.0 ml. Centrifuge 40 ml of this solution and dilute chromatogram obtained with test solution (b) corresponds to
10.0 ml of the clear supernatant liquid to 100.0 ml with 0.01 M the peak in the chromatogram obtained with reference solution
methanolic hydrochloric acid. Measure the absorbance of (c).
the resulting solution at the maximum at about 297 nm (2.4.7).
Calculate the content of C19H21NO taking 150 as the specific D. Dissolve 10 mg in 0.5 ml of nitric acid, add 0.5 ml of water
absorbance at 297 nm. and heat over a flame for 2 minutes. Allow to cool and add
0.5 ml of silver nitrate solution; a white precipitate is
Storage. Store protected from light and moisture. produced.
equivalent amount of doxepin. Tests
Doxorubicin Hydrochloride Related substances. Determine by liquid chromatography
(2.4.14).
O OH O Test solution (a). Dissolve 50 mg of the substance under
OH examination in sufficient of the mobile phase to produce 50 ml.
OH Test solution (b). Dilute 10 ml of test solution (a) to 100 ml
,HCl with the mobile phase.
H3 CO O OH O Reference solution (a). Dissolve 10 mg of doxorubicin
O hydrochloride RS and 10 mg of epirubicin hydrochloride RS
CH3
in the mobile phase and dilute to 50 ml with the mobile phase.
HO Dilute 10 ml of this solution to 100 ml with the mobile phase.
NH2
C27H29NO11,HCl Mol. Wt. 580.0 20 ml with the mobile phase.
Doxorubicin Hydrochloride is (8S,10S)-10-[(3-amino-2,3, 6- Reference solution (c). Dissolve 50 mg of doxorubicin
trideoxy-α -L-lyxo-hexopyranosyl)oxy]-6,8,11-trihydroxy- 8- hydrochloride RS in the mobile phase and dilute to 50 ml with
hydroxyacetyl-1-methoxy-7,8,9,10-tetrahydronaphthacene- the mobile phase. Dilute 10 ml of this solution to 100 ml with
5,12-dione hydrochloride, a substance produced by the the mobile phase.
growth of certain strains of Streptomyces coeruleorubidus Chromatographic system
or S. peucetius or obtained by any other means. – a stainless steel column 25 cm x 4 mm, packed with
Doxorubicin Hydrochloride contains not less than 98.0 per octadecylsilyl silica gel (5 µm),
cent and not more than 102.0 per cent of C27H29NO11,HCl, – mobile phase: a mixture of equal volumes of acetonitrile
calculated on the anhydrous and solvent-free basis. and a solution containing 2.88 g per litre of sodium
441
DOXORUBICIN INJECTION IP 2007
dodecyl sulphate and 2.25 g per litre of phosphoric Doxorubicin Hydrochloride intended for use in the
acid, manufacture of parenteral preparations without a further
– flow rate. 1 ml per minute, appropriate sterilisation procedure complies with the
– spectrophotometer set at 254 nm, following additional requirement.
– a 20 µl loop injector. Sterility (2.2.11). Complies with the test for sterility.
Inject test solution (a) and reference solutions (a) and (b).
Storage. Store protected from moisture. If the material is sterile,
Continue the chromatography for 3.5 times the retention time
it should be stored in sterile, tamper-evident containers and
of doxorubicin of about 8 minutes. The test is not valid unless
in the chromatogram obtained with reference solution (a) the
resolution factor between the peaks due to doxorubicin and Labelling. The label states whether or not the material is
epirubicin is at least 2.0. intended for use in the manufacture of parenteral preparations.
In the chromatogram obtained with the test solution the area
of any peak other than the principal peak is not more than the
area of the peak corresponding to doxorubicin in the Doxorubicin Injection
cent). Ignore any peak with an area less than 0.1 times the area Doxorubicin Hydrochloride Injection
of the peak corresponding to doxorubicin in the chromatogram Doxorubicin Injection is a sterile solution of Doxorubicin
obtained with reference solution (b) (0.5 per cent). Hydrochloride in Water for Injections made isotonic with
Acetone and ethanol. Not more than 2.0 per cent w/w together Sodium Chloride, Dextrose or other suitable added
of which not more than 0.5 per cent w/w is acetone, determined substances. It is either supplied as preformed solution or it is
by gas chromatography (2.4.13), injecting 1 µl of each of two prepared by dissolving the contents of a sealed container
solutions containing Doxorubicin Hydrochloride with or without auxiliary
substances in the requisite amount of Water for Injections or
Test solution. A 5.0 per cent w/v solution of the substance Sodium Chloride Injection as directed on the label.
under examination and 0.1 per cent w/v of dioxon in water.
Doxorubicin Injection contains not less than 90.0 per cent and
Reference solution. 0.05 per cent w/v of acetone, not more than 115.0 per cent of the stated amount of
0.05 per cent w/v of ethanol and 0.1 per cent w/v of the internal doxorubicin hydrochloride, C27H29NO11,HCl.
standard.
– a glass column 2 m x 3 mm, packed with acid-washed
diatomaceous support (180 to 250 mesh) impregnated
with 10 per cent w/w of polyethylene glycol 20,000
(such as Carbowax 20M or Chromosorb E/AW),
– temperature: Tests
column.70°,
inlet port and detector. 125°, pH (2.4.24). 2.5 to 4.5 for the preformed solution and 4.5 to 6.5,
– flow rate. 30 ml per minute of the carrier gas. determined in the injection prepared in accordance with the
directions on the label.
Bacterial endotoxins (2.2.3). Not more than 2.2 Endotoxin Units
Assay. Determine by liquid chromatography (2.4.14) by the
per mg of doxorubicin hydrochloride, determined in a solution
procedure described under the test for Related substances.
prepared by diluting the injection, if necessary, with water
Inject test solution (b) and reference solution (c). BET to obtain a concentration of 2.0 mg of doxorubicin
Calculate the content of C27H29NO11,HCl. hydrochloride per ml.
Doxorubicin Hydrochloride intended for use in the Sterility. Complies with the test for sterility, Method A, (2.2.11),
manufacture of parenteral preparations without a further using the entire contents of all the containers collected
appropriate procedure for the removal of bacterial aseptically.
endotoxins complies with the following additional Assay. Determine by liquid chromatography (2.4.14).
requirement.
Test solution. A solution containing 0.05 per cent w/v of
Bacterial endotoxins (2.2.3). Not more than 2.2 Endotoxin doxorubicin hydrochloride prepared by diluting an accurately
Units per mg. measured volume of the injection containing not less than
442
IP 2007 DOXYCYCLINE HYDROCHLORIDE
2 mg of Doxorubicin Hydrochloride with the mobile phase or carboxamide hydrochloride hemiethanolate hemihydrate, an
by dissolving the contents of the sealed container in sufficient antimicrobial substance obtained from oxytetracycline or
mobile phase to give a solution of the same strength. methacycline or by any other means.
Reference solution (a). A 0.05 per cent w/v solution of Doxycycline Hydrochloride has a potency not less than
doxorubicin hydrochloride RS in the mobile phase. 880 µg and not more than 940 µg of C22H24N2O8 per mg,
calculated on the anhydrous and ethanol-free basis.
w/v each of doxorubicin hydrochloride RS and epirubicin Description. A yellow, crystalline powder; odour, slightly
hydrochloride RS in the mobile phase. ethanolic; hygroscopic.
octadecylsilyl silica gel (5 µm), Test A may be omitted if tests B, C and D are carried out. Tests
– mobile phase: a mixture of 50 volumes of a solution B and C may be omitted if tests A and D are carried out.
containing 0.288 per cent w/v of sodium dodecyl
sulphate and 0.23 per cent w/v of phosphoric acid,
Compare the spectrum with that obtained with doxycycline
45 volumes of acetonitrile and 5 volumes of methanol,
doxycycline hydrochloride.
– a 20 µl loop injector. B. Determine by thin-layer chromatography (2.4.7), coating
the plate with silica gel H.
Inject reference solution (b) six times. The assay is not valid
unless the resolution factor between the peaks due to Mobile phase. A mixture of 59 volumes of dichloromethane,
doxorubicin and epirubicin is not less than 2.0 and the relative 35 volumes of methanol and 6 volumes of water.
standard deviation for the area of the peak due to doxorubicin Test solution. Dissolve 50 mg of the substance under
is not more than 1.0 per cent. If these requirements are not examination in 100 ml of methanol.
met, adjust the operating conditions.
Calculate the content of C27H29NO11,HCl in the injection. doxycycline hydrochloride RS in methanol.
Storage. Store the sealed container at a temperature not Reference solution (b). A solution containing 0.05 per cent
exceeding 30°. Store the preformed solution protected from w/v each of doxycycline hydrochloride RS and tetracycline
light in a refrigerator. Use the solution prepared in the liquid hydrochloride RS in methanol.
stated on the label immediately after preparation but, in any
case, within the period recommended by the manufacturer Spray the plate evenly with a 10 per cent w/v solution of
when prepared and stored strictly in accordance with the disodium edetate the pH of which has been adjusted to
instructions of the manufacturer. 9.0 with 10 M sodium hydroxide. Allow the plate to dry in a
horizontal position for at least 1 hour. Immediately before use
dry it at 110° for 1 hour. Apply to the plate 1 µl of each solution.
After development, dry the plate in a current of air and examine
Doxycycline Hydrochloride in ultraviolet light at 365 nm. The principal spot in the
Doxycycline Hyclate that in the chromatogram obtained with reference solution (a).
OH O OH O reference solution (b) shows two clearly separated spots.
OH
CONH2
C. To about 2 mg add 5 ml of sulphuric acid; a yellow colour
,HCl, 1/2 C 2H5OH, 1/2 H2O is produced.
OH
H H
CH 3 OH N(CH3)2 D. A 5 per cent w/v solution gives the reactions of chlorides
(2.3.1).
Tests
C22H24N2O8,HCl,½C2H6O,½H2O Mol. Wt. 513.0
Doxycycline Hydrochloride is (4S,4aR,5S,5aR,6R,12aS)-4- pH (2.4.24). 2.0 to 3.0, determined in a 1.0 per cent w/v solution.
dimethylamino-1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12, 12a- Specific optical rotation (2.4.22). -105° to -120°, determined
pentahydroxy-6-methyl-1,11-dioxonaphthacene-2- within 5 minutes of preparing, in a 1.0 per cent w/v solution in
443
DOXYCYCLINE HYDROCHLORIDE IP 2007
a mixture of 1 volume of 1 M hydrochloric acid and 99 volumes unless (a) the resolution factor between the first peak
of methanol. (methacycline) and the second peak (6-epidoxycycline) is at
Light absorption (2.4.7). Absorbance of a 0.001 per cent w/v least 1.25, (b) the resolution factor between the second peak
solution in a mixture of 1 volume of 1 M hydrochloric acid and the third peak (doxycycline) is at least 2.0 (adjust the
and 99 volumes of methanol, measured within 1 hour of content of 2-methyl-2-propanol in the mobile phase if necessary)
preparing the solution, at the maximum at about 349 nm, and (c) the symmetry factor for the third peak is at most 1.25.
0.300 to 0.335. Inject reference solution (a) six times. The test is not valid
Light-absorbing impurities. Dissolve 0.1 g in sufficient of a unless the relative standard deviation of the area of the peak
mixture of 1 volume of 1 M hydrochloric acid and 99 volumes due to doxycycline is not more than1.0 per cent.
of methanol to produce 10 ml. Absorbance of the solution at Inject the test solution and reference solution (d). In the
about 490 nm, when measured within 1 hour of preparing the chromatogram obtained with the test solution the area of any
solution, not more than 0.07 (2.4.7). peak corresponding to methacycline or 6-epidoxycycline is
Related substances. Determine by liquid chromatography not greater than the area of the corresponding peak in the
(2.4.14). chromatogram obtained with reference solution (d); the area
of any peak appearing between the solvent peak and the peak
Test solution. Dissolve 80 mg of the substance under corresponding to methacycline and the area of any peak
examination in 100 ml of 0.01 M hydrochloric acid. appearing on the tail of the main peak is not greater than
Reference solution (a). A 0.08 per cent w/v solution of 25 per cent of that of the peak corresponding to
doxycycline hydrochloride RS in 0.01 M hydrochloric acid. 6-epidoxycycline in the chromatogram obtained with reference
solution (d).
6-epidoxycycline hydrochloride RS in 0.01 M hydrochloric Ethanol. 4.3 to 6.0 per cent w/w of C2H6O, determine by gas
acid. chromatography (2.4.13).
Reference solution (c). A 0.08 per cent w/v solution of Test solution (a). A 1 per cent w/v solution of the substance
methacycline hydrochloride RS in 0.01 M hydrochloric acid. under examination in a 0.05 per cent v/v solution of 1-propanol
Reference solution (d). A solution containing 0.0016 per cent (internal standard) in water (solution A).
w/v each of 6-epidoxyccline hydrochloride RS and Test solution (b). A 1 per cent w/v solution of the substance
methacycline hydrochloride RS in 0.01 M hydrochloric acid. under examination in water.
Reference solution (e). Dilute a mixture of 4 volumes of Reference solution. A 0.05 per cent v/v solution of ethanol in
reference solution (a), 1.5 volumes of reference solution (b) solution A.
and 1 volume of reference solution (c) to 25 volumes with
0.01 M hydrochloric acid.
– a column 1.5m x 4 mm, packed with porous polymer
Chromatographic system beads (80 to 100 mesh) (such as Porapak Q),
– a stainless steel column 25 cm x 4.6 mm, packed with – temperature:
styrene-divinylbenzene co-polymer (8 to10 µm), column.135°,
– mobile phase: a solution prepared by adding 60 g of inlet port and detector. 150°,
2-methyl-2-propanol to a volumetric flask with the aid
Calculate the content of C2H6O taking 0.790 g as its weight per
of 200 ml of water, adding 400 ml of phosphate buffer
ml at 20°.
pH 8.0, 50 ml of a 1 per cent w/v solution of tetrabutyl
ammonium hydrogen sulphate previously adjusted to Heavy metals (2.3.13). 0.4 g complies with the limit test for
pH 8.0 with 2 M sodium hydroxide and 10 ml of a 4 per heavy metals, Method B (50 ppm).
cent w/v solution of disodium edetate previously
adjusted to pH 8.0 with 2 M sodium hydroxide and
diluting to 1 litre with water, Water (2.3.43). 1.4 to 2.8 per cent, determined on 1.2 g.
– column temperature: 60° Assay. Determine by liquid chromatography (2.4.14).
– spectrophotometer set at 254 nm, Test solution. Weigh accurately 80 mg of the substance under
– a 20 µl loop injector. examination and dissolve in 100 ml of 0.01 M hydrochloric
acid.
Using reference solution (e) adjust the attenuation to obtain
peaks with a height corresponding to at least 50 per cent of Reference solution. A 0.08 per cent w/v solution of doxycycline
full-scale deflection on the chart paper. The test is not valid hydrochloride RS in 0.01 M hydrochloric acid.
444
IP 2007 DOXYCYCLINE CAPSULES

styrene-divinylbenzene co-polymer (8 to 10 µm), A. Determine by thin-layer chromatography (2.4.17), coating
– column temperature: 60° the plate with silica gel H.
– mobile phase: a solution prepared by adding 60 g of Mobile phase. A mixture of 59 volumes of dichloromethane,
2-methyl-2-propanol to a volumetric flask with the aid 35 volumes of methanol and 6 volumes of water.
of 200 ml of water, adding 400 ml of phosphate buffer Test solution. Shake a quantity of the contents of the capsules
pH 8.0, 50 ml of a 1 per cent w/v solution of tetrabutyl containing 50 mg of anhydrous doxycycline with 100 ml of
ammonium hydrogen sulphate previously adjusted to methanol for 1 to 2 minutes, centrifuge and use the supernatant
pH 8.0 with 2 M sodium hydroxide and 10 ml of a 4 per liquid. Prepare freshly.
cent w/v solution of disodium edetate previously
adjusted to pH 8.0 with 2 M sodium hydroxide and Reference solution (a). A 0.05 per cent w/v solution of
diluting to 1 litre with water, doxycycline hydrochloride RS in methanol.
– flow rate. 1 ml per minute, Reference solution (b). A solution containing 0.05 per cent
– spectrophotometer set at 254 nm, w/v each of doxycycline hydrochloride RS and tetracycline
– a 20 µl loop injector. hydrochloride RS in methanol.
Inject the reference solution six times. The test is not valid Spray the plate evenly with a 10 per cent w/v solution of
unless the relative standard deviation of the area of the peak disodium edetate the pH of which has been adjusted to
due to doxycycline is not more than 1.0 per cent. 9.0 with 10 M sodium hydroxide. Allow the plate to dry in a
horizontal position for at least 1 hour. Immediately before use
dry it at 110° for 1 hour. Apply to the plate 1 µl of each solution.
Calculate the content of C22H24N2O8. After development, dry the plate in a current of air and examine
Doxycycline Hydrochloride intended for use in the it in ultraviolet light at 365 nm. The principal spot in the
manufacture of parenteral preparations without a further chromatogram obtained with the test solution corresponds to
appropriate procedure for the removal of bacterial that in the chromatogram obtained with reference solution (a).
endotoxins complies with the following additional The test is not valid unless the chromatogram obtained with
requirement. reference solution (b) shows two clearly separated spots.
B. To 0.5 mg of the contents of the capsules add 2 ml of
sulphuric acid; a yellow colour is produced.
Units per mg.
C. A 5 per cent w/v solution of the contents of the capsules
Doxycycline Hydrochloride intended for use in the
gives the reactions of chlorides (2.3.1).
appropriate sterilisation procedure complies with the Tests
Light-absorbing impurities. Dissolve the contents of
Sterility (2.2.11). Complies with the test for sterility. 5 capsules as completely as possible in sufficient of a mixture
Storage. Store protected from light and moisture at a of 1 volume of 1 M hydrochloric acid and 99 volumes of
temperature not exceeding 30°. If the substance is intended methanol to produce a solution containing the equivalent of
for use in the manufacture of parenteral preparations, the 1.0 per cent w/v of anhydrous doxycycline and filter.
container should be sterile, tamper-evident and sealed so as Absorbance of the filtrate at about 490 nm, not greater than
to exclude micro-organisms. 0.2 (2.4.7), calculated with reference to the dried contents of
the capsules.
Labelling. The label states, where applicable, that the material
is sterile.
(2.4.14).
Test solution. Dissolve a quantity of the contents of the
capsules containing 7 mg of anhydrous doxycycline in 10 ml
Doxycycline Capsules of 0.01 M hydrochloric acid, filter and use the filtrate.
Doxycycline Hydrochloride Capsules doxycycline hydrochloride RS in 0.01 M hydrochloric acid.
Doxycycline Capsules contain not less than 90.0 per cent and Reference solution (b). A 0.08 per cent w/v solution of
not more than 120.0 per cent of the stated amount of 6-epidoxycycline hydrochloride RS in 0.01 M hydrochloric
doxycycline, C22H24N2O8. acid.
445
DYDROGESTERONE IP 2007
Reference solution (c). A 0.08 per cent w/v solution of Loss on drying (2.4.19). Not more than 8.5 per cent, determined
methacycline hydrochloride RS in 0.01 M hydrochloric acid. on 0.5 g of the contents of the capsules by drying in an oven
at 105° for 2 hours.
w/v each of 6-epidoxyccline hydrochiloride RS and Other tests. Comply with the tests stated under Capsules.
methacycline hydrochloride RS in 0.01 M hydrochloric Assay. Determine by liquid chromatography (2.4.14).
acid.
Test solution. Dissolve the mixed contents of 20 capsules
Reference solution (e). Dilute a mixture of 4 volumes of containing about 17.5 mg of anhydrous doxycycline in
reference solution (a), 1.5 volumes of reference solution (a) sufficient 0.01 M hydrochloric acid to produce 25.0 ml and
and 1 volume of reference solution (c) to 25 volumes with dilute 4.0 ml of this solution to 25.0 ml with the same solvent.
0.01 M hydrochloric acid. Reference solution. A 0.0128 per cent w/v solution of
Chromatographic system doxycycline hydrochloride RS in 0.01 M hydrochloric acid.
styrene-divinylbenzene co-polymer (8 to 10 µm), – a stainless steel column 25 cm x 4.6 mm, packed with
– column temperature 60° styrene-divinylbenzene co-polymer (8 to 10 µm),
– mobile phase: a solution prepared by adding 60 g of – column temperature 60°,
2-methyl-2-propanol to a volumetric flask with the aid – mobile phase: a solution prepared by adding 60 g of 2-
of 200 ml of water, adding 400 ml of phosphate buffer methyl-2-propanol to a volumetric flask with the aid of
pH 8.0, 50 ml of a 1 per cent w/v solution of tetrabutyl 200 ml of water, adding 400 ml of phosphate buffer pH
ammonium hydrogen sulphate previously adjusted to 8.0, 50 ml of a 1 per cent w/v solution of tetrabutyl
pH 8.0 with 2 M sodium hydroxide and 10 ml of a 4 per ammonium hydrogen sulphate previously adjusted to
cent w/v solution of disodium edetate previously pH 8.0 with 2 M sodium hydroxide and 10 ml of a 4 per
adjusted to pH 8.0 with 2 M sodium hydroxide and cent w/v solution of disodium edetate previously
diluting to 1000 ml with water, adjusted to pH 8.0 with 2 M sodium hydroxide and
– flow rate. 1 ml per minute, diluting to 1 litre with water,
– spectrophotometer set at 254 nm, – flow rate. 1 ml per minute,
– a 20 µl loop injector. – spectrophotometer set at 254 nm,
Using reference solution (e) adjust the attenuation to obtain – a 20 µl loop injector.
peaks with a height corresponding to at least 50 per cent of Inject the reference solution six times. The test is not valid
full-scale deflection of the recorder. The test is not valid unless unless the relative standard deviation of the area of the peak
(a) the resolution factor between the first peak (methacycline) due to doxycycline is not more than1.0 per cent.
and the second peak (6-epidoxycycline) is at least 1.25, (b) the Inject alternately the test solution and the reference solution.
resolution factor between the second peak and the third peak
Calculate the content of C22H24N2O8 in the capsules.
(doxycycline) is at least 2.0 (adjust the content of
2-methylpropan-2-ol in the mobile phase if necessary). Storage. Store protected from light and moisture at a
Inject reference solution (a).The test is not valid unless the
relative standard deviation for replicate injections is not more Labelling. The label states the strength in terms of the
than 2.0 per cent. equivalent amount of doxycycline.
Inject the test solution and reference solution (d) and record
the chromatograms. In the chromatogram obtained with the Dydrogesterone
test solution the area of any peak corresponding to
methacycline or 6- epidoxycycline is not greater than the area O
of the corresponding peak in the chromatogram obtained with H3 C CH3
reference solution (d) (2 per cent, with reference to doxycycline
hydrochloride), the area of any peak appearing between the H3C H
solvent peak and the peak corresponding to methacycline
and the area of any peak appearing on the tail of the main peak H H
is not greater than 25 per cent of that of the peak corresponding O
to 6-epidoxycycline in the chromatogram obtained with
reference solution (d) (0.5 per cent, with reference to C21H28O2 Mol. Wt. 312.5
doxycycline hydrochloride). Dydrogesterone is 9β,10α-pregna-4,6-diene-3,20-dione.
446
IP 2007 DYDROGESTERONE TABLETS
Dydrogesterone contains not less than 97.0 per cent and not dydrogesterone), 1.0 and impurity C (∆ 8-14 triene
more than 103.0 per cent of C21H28O2, calculated on the dried dydrogesterone), 0.899.
basis. The test is not valid unless the column efficiency is not less
Description. A white or almost white, crystalline powder; than 10000 theoretical plates in the chromatogram obtained
odourless or almost odourless. with the test solution.
For impurity A and impurity B, spectrophotometer set at
Identification
280 nm.
A. Determine by infrared absorption spectrophotometry (2.4.6). Inject the test solution and reference solution (b). In the
Compare the spectrum with that obtained with dydrogesterone chromatogram obtained with the test solution the area of any
RS or with the reference spectrum of dydrogesterone. peak corresponding to impurity A is not more than the area of
B. In the Assay, the principal peak in the chromatogram the principal peak in the chromatogram obtained with reference
obtained with the test solution corresponds to the peak in the solution (b) (0.2 per cent) and the area of any peak
chromatogram obtained with the reference solution. corresponding to impurity B is not more than 2.5 times the
Tests reference solution (b) (0.5 per cent).
Specific optical rotation (2.4.22). –446º to –464º, determined For impurity C, spectrophotometer set at 385 nm:
in a 1.0 per cent w/v solution in dioxan. Inject the test solution and reference solution (b). In the
Light absorption (2.4.7). When examined in the range 230 nm chromatogram obtained with the test solution the area of any
to 360 nm, the final solution obtained in the Assay shows an peak corresponding to impurity C, using the response factor,
absorption maximum only at about 286 nm. The ratio of the is not more than the area of the principal peak in the
absorbance at about 240 nm to that at about 286 nm is not chromatogram obtained with reference solution (b) (0.5 per
more than 0.12. cent).
Related substances. Determine by liquid chromatography The area of any other individual impurity is not more than
(2.4.14). 0.5 times the area of the principal peak in the chromatogram
examination in the mobile phase and dilute to 100.0 ml with the The sum of the areas of all the impurities is not greater than
mobile phase. 5 times the area of the principal peak in the chromatogram
Reference solution (a). Dissolve 20 mg of dydrogesterone RS
in the mobile phase and dilute to 100 ml with the mobile phase. Sulphated ash (2.3.18). Not more than 0.1 per cent, determined
on 2.0 g.
Reference solution (b). Dilute 1 ml of the test solution to 500
ml with the mobile phase. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
on 1.0 g by drying in an oven at 105º for 3 hours.
Assay. Determine by liquid chromatography (2.4.14) as
– a stainless steel column 15 cm x 4.6 mm, packed with described under Related substances.
– column. temperature 40º, Inject alternately the test solution and reference solution (a).
– mobile phase: a mixture of 53 volumes of water with 26 Calculate the content of C21H28O2.
volumes of ethanol (95 per cent) and 21 volumes of Storage. Store protected from light and moisture.
acetonitrile,
– spectrophotometer set at 280 nm and 385 nm,
– a 10 µl loop injector. Dydrogesterone Tablets
The relative retention time with respect to dydrogesterone Dydrogesterone Tablets contain not less than 92.5 per cent
(retention time, about 10.5 minutes): impurity A and not more than 107.5 per cent of the stated amount of
(6-dehydroprogesterone), about 1.16, impurity B (17α- dydrogesterone, C21H28O2.
dydrogesterone) about 1.32 and impurity C (∆ 8-14 triene
dydrogesterone), about 0.97. Identification
The response factors relative to that of dydrogestrone: Extract a quantity of the powdered tablets containing 60 mg of
impurity A (6-dehydroprogesterone) 1.0, impurity B (17α− Dydrogesterone with 20 ml of methanol, filter and evaporate
447
DYDROGESTERONE TABLETS IP 2007
the filtrate to dryness. The residue complies with the following peaks corresponding to 6-dehydroprogesterone and 17α-
tests. dydrogesterone are not more than the area of the principal
A. Determine by infrared absorption spectrophotometry (2.4.6). peak obtained with reference solution (b) (0.5 per cent each)
Compare the spectrum with that obtained with dydrogesterone and the sum of all impurities found is not more than twice the
RS or with the reference spectrum of dydrogesterone. area of the principal peak obtained with reference solution (b)
(1.0 per cent).
Tablets.
Determine by liquid chromatography (2.4.14) as given under
Tests Related substances using the following test solution.
Related substances. Determine by liquid chromatography Test solution. To one tablet add 5 ml of water and mix with the
(2.4.14). aid of ultrasound. Add 40 ml of acetonitrile and mix with the
aid of ultrasound for 15 minutes. Add about 50 ml of water
Test solution. Determine the average weight of 20 tablets. Do
and swirl for 15 minutes. Dilute with sufficient water to
not powder the tablets. Accurately weigh 20 tablets. Add
produce 100.0 ml.
25 ml of water and mix with aid of ultrasound until complete
disintegration has occurred. Add 200 ml of acetonitrile and Other tests. Comply with the tests stated under Tablets.
mix with aid of ultrasound for 15 minutes, dilute to 500.0 ml Assay. Determine by liquid chromatography (2.4.14) as
with water and mix. Centrifuge a part of this solution in a described under Related substances.
closed tube for about 5 minutes. Dilute a suitable volume of
the supernatant liquid with the mobile phase to obtain a Inject alternately the test solution and reference solution (a).
solution containing about 0.1 mg of dydrogesterone per ml Calculate the content of C21H28O2 in the tablets.
and mix. Filter a part of this solution, discarding the first few
ml of the filtrate. Storage. Store protected from light.
Reference solution (a). Weigh accurately about 10 mg of

dydrogesterone RS, add 40 ml of acetonitrile and mix with aid
of ultrasound and dilute to 100.0 with water,
Reference solution (b). Dilute 1 ml of the test solution to 200.0
– mobile phase: a mixture of 600 volumes of water and 425
volumes of acetonitrile,
The relative retention time with respect to dydrogesterone
(retention time, about 10.5 minutes); impurity A (6-
dehydroprogesterone), about 1.16 and impurity B (17α−
dydrogesterone), about 1.32.
The response factors relative to dydrogestrone are 0.74 for
impurity A and 1.0 for impurity B.
The test is not valid unless the column efficiency is not less
than 10000 theoretical plates in the chromatogram obtained
with test solution.
448
E
Econazole Nitrate ....
Econazole Cream ....
Econazole Pessaries ....
Efavirenz ....
Efavirenz Capsules ....
Efavirenz Tablets ....
Emetine Hydrochloride ....
Emetine Injection ....
Emtricitabine ....
Emtricitabine Capsules ....
Emulsifying Wax ....
Enalapril Maleate ....
Enalapril Maleate Tablets ....
Ephedrine ....
Ephedrine Hydrochloride ....
Ephedrine Oral Solution ....
Ephedrine Tablets ....
Ergocalciferol ....
Ergometrine Maleate ....
Ergometrine Injection ....
Ergometrine Tablets ....
Ergotamine Tartrate ....
Ergotamine Injection ....
Ergotamine Tablets ....
Erythromycin ....
Erythromycin Tablets ....
Erythromycin Estolate ....
Erythromycin Estolate Tablets ....
Erythromycin Stearate ....
Erythromycin Stearate Tablets ....
449
Ethacrynic Acid ....

Ethacrynic Acid Tablets ....
Ethambutol Hydrochloride ....
Ethambutol Hydrochloride Tablets ....
Ethambutol And Isoniazid Tablets ....
Ethanol ....
Ethanol (95 Per Cent) ....
Anaesthetic Ether ....
Ethinyloestradiol ....
Ethinyloestradiol Tablets ....
Ethionamide ....
Ethionamide Tablets ....
Ethopropazine Hydrochloride ....
Ethopropazine Tablets ....
Ethosuximide ....
Ethosuximide Capsules ....
Ethosuximide Syrup ....
Ethylcellulose ....
Ethyl Chloride ....
Ethyl Oleate ....
Ethylenediamine Hydrate ....
Ethyloestrenol ....
Ethyloestrenol Tablets ....
Etoposide ....
Etoposide Capsules ....
Etoposide Injection ....
Etoposide Concentrate ....
450
IP 2007 ECONAZOLE CREAM
Solvent mixture. A mixture of 9 volumes of methanol and 1

Econazole Nitrate volume of strong ammonia solution.
Cl examination in 10 ml of the solvent mixture.
O Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with

N the solvent mixture.
, HNO3
N Reference solution (a). Dilute 5 ml of test solution (b) to 200
ml with the solvent mixture.
Cl Reference solution (b). A 0.5 per cent w/v solution of
Cl econazole nitrate RS in the solvent mixture.
C18H15Cl3N2O,HNO3 Mol. Wt. 444.7 phase to rise 10 cm. Dry the plate in a current of air for
Econazole Nitrate is (RS)-1-[2-(4-chlorophenylmethoxy)-2- 15 minutes and examine in ultraviolet light at 254 nm. Any
(2,4-dichlorophenyl)ethyl]-1H-imidazole nitrate. secondary spot in the chromatogram obtained with test
Econazole Nitrate contains not less than 98.5 per cent and not chromatogram obtained with reference solution (a). Spray the
more than 101.5 per cent of C18H15Cl3N2O,HNO3, calculated on plate with modified potassium iodobismuthate solution and
the dried basis. examine in daylight. Any secondary spot in the chromatogram
Description. A white or almost white, crystalline powder. obtained with test solution (a) is not more intense than the
Identification (a). The test is not valid unless a spot is clearly visible in the
Compare the spectrum with that obtained with econazole
nitrate RS or with the reference spectrum of econazole nitrate. Assay. Weigh accurately about 0.4 g and dissolve in 50 ml of
B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.04
per cent w/v solution in a mixture of 1 volume of 0.1 M
hydrochloric acid and 9 volumes of 2-propanol shows
absorption maxima at about 265 nm, 271 nm and 280 nm; the 1 ml of 0.1 M perchloric acid is equivalent to 0.04447 g of
ratio of the absorbance at the maximum at about 271 nm to that C18H15Cl3N2O,HNO3.
at the maximum at about 280 nm is 1.55 to 1.70. Storage. Store protected from light.
C. In the test for Related substances examine the
chromatograms obtained in ultraviolet light at 254 nm before
spraying. The principal spot in the chromatogram obtained
with test solution (b) corresponds to that in the chromatogram Econazole Cream
Econazole Nitrate Cream
D. Gives reaction A of nitrates (2.3.1).
Econazole Cream contains Econazole Nitrate in a suitable basis.
Tests Econazole Cream contains not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of econazole
Appearance of solution. A 1.0 per cent w/v solution in nitrate, C18H15Cl3N2O,HNO3.
than reference solution YS7 (2.4.1). Identification
Related substances. Determine by thin-layer chromatography A. Mix a quantity of the cream containing 40 mg of Econazole
(2.4.17), coating the plate with silica gel GF254. Nitrate with 20 ml of a mixture of 1 volume of 1 M sulphuric
Mobile phase. A mixture of 60 volumes of dioxan, 40 volumes acid and 4 volumes of methanol and shake with two quantities,
of toluene and 1 volume of strong ammonia solution. each of 50 ml, of carbon tetrachloride, discarding the organic
451
ECONAZOLE PESSARIES IP 2007
layers. Make the aqueous phase alkaline with 2 M ammonia Calculate the content of C18H15Cl3N2O,HNO3 in the cream.
and extract with two quantities, each of 40 ml, of chloroform. Storage. Store protected from light at a temperature not
Combine the chloroform extracts, shake with 5 g of anhydrous exceeding 30°. If it is packed in aluminium tubes the inner
sodium sulphate, filter and dilute the filtrate to 100 ml with surfaces of the tubes should be coated with a suitable lacquer.
chloroform. Evaporate 50 ml to dryness and dissolve the
residue in 50 ml of a mixture of 1 volume of 0.1 M hydrochloric
acid and 9 volumes of 2-propanol.
When examined in the range 230 nm to 360 nm (2.4.7), the Econazole Pessaries
resulting solution shows absorption maxima at about 265 nm, Econazole Nitrate Pessaries; Econazole Vaginal Tablets
271 nm and 280 nm. The ratio of the absorbance at about
271 nm to that at about 280 nm is 1.55 to 1.70. Econazole Pessaries contain Econazole Nitrate in a suitable
basis.
obtained with the test solution corresponds to the peak due Econazole Pessaries contain not less than 90.0 per cent and
to econazole in the chromatogram obtained with the reference not more than 110.0 per cent of the stated amount of econazole
solution (a). nitrate, C18H15Cl3N2O,HNO3.
Identification
Tests
A. Mix a quantity of the crushed pessaries containing 40 mg
Other tests. Complies with the tests stated under Creams. of Econazole Nitrate with 20 ml of a mixture of 1 volume of 1 M
Assay. Determine by gas chromatography (2.4.13). sulphuric acid and 4 volumes of methanol and shake with
two quantities, each of 50 ml, of carbon tetrachloride,
Test solution. Mix a quantity of the cream containing about 40
discarding the organic layers. Make the aqueous phase alkaline
mg of Econazole Nitrate with 20 ml of a mixture of 1 volume of
with 2 M ammonia and extract with two quantities, each of 40
0.5 M sulphuric acid and 4 volumes of methanol and shake
ml, of chloroform. Combine the chloroform extracts, shake
with two quantities, each of 50 ml, of carbon tetrachloride.
with 5 g of anhydrous sodium sulphate, filter and dilute the
Wash each organic layer in turn with the same 10-ml quantity
filtrate to 100 ml with chloroform. Evaporate 50 ml to dryness
of a mixture of 1 volume of 0.5 M sulphuric acid and 4 volumes
and dissolve the residue in 50 ml of a mixture of 1 volume of
of methanol. Combine the aqueous phase and the washings,
0.1 M hydrochloric acid and 9 volumes of 2-propanol.
make alkaline with 2 M ammonia and extract with two
quantities, each of 50 ml, of chloroform. To the combined When examined in the range 230 nm to 360 nm (2.4.7), the
extracts add 10.0 ml of a 0.3 per cent w/v solution of 1,2,3,4- resulting solution shows absorption maxima at about 265 nm,
tetraphenylcyclopenta-1,3-diene (internal standard) in 271 nm and 280 nm. The ratio of the absorbance at about
chloroform and 5 g of anhydrous sodium sulphate, shake, 271 nm to that at about 280 nm is 1.55 to 1.70.
filter, evaporate the filtrate to a low volume and add sufficient B. In the test for Related substances, the principal spot in the
chloroform to produce 10.0 ml. chromatogram obtained with the test solution corresponds to
Reference solution (a). Shake 40 mg of econazole nitrate RS that in the chromatogram obtained with reference solution
with 10.0 ml of a 0.3 per cent w/v solution of the internal (b).
standard in chloroform and 0.2 ml of strong ammonia solution,
add 1 g of anhydrous sodium sulphate, shake again and filter. Tests
Reference solution (b). Prepare in the same manner as Related substances. Determine by thin-layer chromatography
reference solution (a) but omit the addition of the internal (2.4.17), using a precoated silica gel plate (such as Merck
standard solution. silica gel 60 plates).
Chromatographic system Mobile phase. A mixture of 70 volumes of chloroform, 20
– a glass column 1.5 m x 2 mm, packed with acid-washed, volumes of methanol and 10 volumes of an 85 per cent w/v
silanised diatomaceous support (80 to 100 mesh) coated solution of formic acid.
with 3 per cent w/w of phenyl methyl silicone fluid Test solution. Mix a quantity of the crushed pessaries
(50 per cent phenyl) (such as OV-17), containing 40 mg of Econazole Nitrate with 40 ml of methanol
– temperature: and heat under a reflux condenser for 15 minutes. Allow to
column. 270°, cool, filter, wash the filter paper with methanol and evaporate
inlet port and detector. 300°, the filtrate and washings to a volume of about 5 ml. Filter
– flow rate. 30 ml per minute of the carrier gas. through a filter paper (such as Whatman No. 50 paper), wash
452
IP 2007 EFAVIRENZ
the paper with methanol, evaporate the filtrate and washings B. In the Assay, the principal peak in the chromatogram
to dryness and dissolve the residue in 2 ml of methanol. obtained with the test solution corresponds to the peak in the
Reference solution (a). Dilute 0.5 ml of the test solution to 100 chromatogram obtained with the reference solution.
ml with methanol.
Tests
econazole nitrate RS in methanol. Specific optical rotation (2.4.22). –90.0° to –100.0°, determined
dry the plate in air and expose to iodine vapour for 1 hour. Any Related substances. Determine by liquid chromatography
secondary spot in the chromatogram obtained with the test (2.4.14).
solution is not more intense than the spot in the chromatogram Test solution. Dissolve 0.1 g of the substance under
obtained with reference solution (a). Ignore any spot with an examination in 10 ml of methanol.
Rf value higher than 0.9.
Other tests. Complies with the tests stated under Pessaries. with methanol.
Assay. Dissolve 5 pessaries in 250.0 ml of anhydrous glacial Chromatographic system
acetic acid with the aid of gentle heat and allow to cool. – a stainless steel column 25 cm x 4.6 mm, packed with
Titrate 100.0 ml of the solution with 0.1 M perchloric acid, base deactivated octadecylsilyl silica gel (5 µm),
determining the end-point potentiometrically (2.4.25). Carry – mobile phase: a filtered and degassed mixture of 50
out a blank titration. volumes of acetonitrile and 50 volumes of a 0.86 per
1 ml of 0.1 M perchloric acid is equivalent to 0.04447 g of cent w/v solution of ammonium dihydrogen phosphate,
C18H15Cl3N2O,HNO3. the pH of which is adjusted to 3.0 ± 0.05 with phosphoric
acid,
Storage. Store protected from light. – flow rate. 1.5 ml per minute,
Efavirenz column efficiency determined from the efavirenz peak is not
less than 6000 theoretical plates and the tailing factor is not
H more than 2.0.
N O Inject the test solution. Determine the amount of related
substances by area normalisation method. The content of
O
Cl any individual impurity is not more than 0.5 per cent and the
F3C C sum of all the impurities is not more than 1.0 per cent.
C
C14H9ClF3NO2 Mol. Wt. 315.7 Loss on drying (2.4.19). Not more than 1.0 per cent, determined
Efavirenz is (4S)-6-chloro-4-(cyclopropylethynyl)-1,4- on 1.0 g by drying at 105° in an oven for 3 hours.
dihydro-4-(trifluoromethyl)-2H-3,1-benzoxazin-2-one. Assay. Determine by liquid chromatography (2.4.14).
Efavirenz contains not less than 98.0 per cent and not more Test solution. Dissolve 0.12 g of the substance under
than 102.0 per cent of C14H9ClF3NO2, calculated on the dried examination in 100 ml of methanol.
basis.
Reference solution. A 0.12 per cent w/v solution of efavirenz
Description. A white or almost white powder. RS in methanol.

A. Determine by infrared absorption spectrophotometry (2.4.6). octadecylsilyl silica gel (5 µm),
Compare the spectrum with that obtained with efavirenz RS or – mobile phase: a filtered and degassed mixture of 50
with the reference spectrum of efavirenz. volumes of acetonitrile and 50 volumes of a 0.86 per
453
EFAVIRENZ CAPSULES IP 2007
cent w/v solution of ammonium dihydrogen phosphate, – spectrophotometer set at 252 nm,
the pH of which is adjusted to 3.0 ± 0.05 with phosphoric – a 20 µl loop injector.
acid, Inject the reference solution. The test is not valid unless the
– flow rate. 1.5 ml per minute, column efficiency determined from the efavirenz peak is not
– spectrophotometer set at 254 nm, less than 6000 theoretical plates and the tailing factor is not
– a 20 µl loop injector. more than 2.0.
Inject the reference solution. The test is not valid unless the Inject the test solution. Determine the amount of related
column efficiency determined from the efavirenz peak is not substances by the area normalisation method. The content of
less than 6000 theoretical plates, the tailing factor is not more any individual impurity is not more than 1.0 per cent and the
than 2.0 and the relative standard deviation for replicate sum of all impurities is not more than 2.0 per cent.
Apparatus. No. 1
Calculate the content of C14H9ClF3NO2. Medium. 900 ml of a 1 per cent w/v solution of sodium lauryl
Storage. Store protected from light. sulphate
a membrane filter disc with an average pore diameter not greater
Efavirenz Capsules than 1.0 µm, rejecting the first few ml of the filtrate and dilute
Efavirenz Capsules contain not less than 90.0 per cent and not a suitable volume of the filtrate if necessary, with the same
more than 110.0 per cent of the stated amount of efavirenz, solvent. Measure the absorbance of the resulting solution at
C14H9ClF3NO2. the maximum at about 252 nm (2.4.7). Calculate the content of
efavirenz, C14H9ClF3NO2 in the medium from the absorbance
Identification obtained from a solution of known concentration of efavirenz
RS in the same solvent.
test solution in the Assay shows an absorption maximum at
C14H9ClF3NO2.
about 252 nm.
obtained with the test solution corresponds to the peak in the Assay. Determine by liquid chromatography (2.4.14).
chromatogram obtained with the reference solution. Test solution. Mix well the contents of 20 capsules and shake
a quantity of the mixed contents containing about 60 mg of
Tests Efavirenz with sufficient methanol to obtain a solution
Related substances. Determine by liquid chromatography containing 6 mg of Efavirenz per ml. Disperse the mixture with
(2.4.14). the aid of ultrasound for 20 minutes, filter through a membrane
filter disc with an average pore diameter not greater than 1.0
Test solution. Mix well the contents of 20 capsules and shake µm, rejecting the first few ml of the filtrate, and dilute 10.0 ml of
a quantity of the mixed contents containing about 5 mg of the filtrate to 50.0 ml with methanol.
Efavirenz with sufficient methanol to obtain a solution
Reference solution. A 0.12 per cent w/v solution of efavirenz
containing 1 mg per ml of Efavirenz.
RS in methanol.
Reference solution. A 0.1 per cent w/v solution of efavirenz Chromatographic system
RS in methanol. – a stainless steel column 25 cm x 4.6 mm, packed with
Chromatographic system base deactivated octadecylsilyl silica gel (5 µm),
– a stainless steel column 25 cm x 4.6 mm, packed with – mobile phase: a filtered and degassed mixture of
base deactivated octadecylsilyl silica gel (5 µm), 50 volumes of acetonitrile and 50 volumes of a 0.86 per
– mobile phase: a filtered and degassed mixture of 50 cent w/v solution of ammonium dihydrogen phosphate,
volumes of acetonitrile and 50 volumes of a 0.86 per the pH of which is adjusted to 3.0 ± 0.05 with phosphoric
cent w/v solution of ammonium dihydrogen phosphate, acid,
the pH of which is adjusted to 3.0 ± 0.05 with phosphoric – flow rate. 1.5 ml per minute,
acid, – spectrophotometer set at 252 nm,
– flow rate. 1.5 ml per minute, – a 20 µl loop injector.
454
IP 2007 EFAVIRENZ TABLETS
Inject the reference solution. The test is not valid unless the Inject the test solution. Determine the amount of related
column efficiency determined from the efavirenz peak is not substances by the area normalisation method. The content of
less than 6000 theoretical plates, the tailing factor is not more any individual impurity is not more than 1.0 per cent and the
than 2.0 and the relative standard deviation for replicate sum of all impurities is not more than 2.0 per cent.
injections is not more than 2.0 per cent. Dissolution (2.5.2).
Apparatus. No. 1
Calculate the content of C14H9ClF3NO2. Medium. 900 ml of a 1 per cent w/v solution of sodium lauryl
sulphate
Efavirenz Tablets a membrane filter disc with an average pore diameter not greater
Efavirenz Tablets contain not less than 90.0 per cent and not than 1.0 µm, reject the first few ml of the filtrate and dilute a
more than 110.0 per cent of the stated amount of efavirenz, suitable volume of the filtrate if necessary, with the same
C14H9ClF3NO2. solvent. Measure the absorbance of the resulting solution at
Identification efavirenz, C14H9ClF3NO2 in the medium from the absorbance
obtained from a solution of known concentration of efavirenz
A. When examined in the range 220 nm to 350 nm (2.4.7), the RS in the same solvent.
test solution in the Assay shows an absorption maximum at
about 252 nm. D. Not less than 70 per cent of the stated amount of
C14H9ClF3NO2.
obtained with the test solution corresponds to the peak in the Other tests. Comply with the tests stated under Tablets.
chromatogram obtained with the reference solution. Assay. Determine by liquid chromatography (2.4.14).
Tests Test solution. Weigh and powder 20 tablets. Weigh accurately

a quantity of the powdered tablets containing about 100 mg
Related substances. Determine by liquid chromatography of Efavirenz and shake with sufficient methanol to obtain a
(2.4.14). mixture containing 6 mg of Efavirenz per ml. Disperse the
Test solution. Shake a quantity of the powdered tablets with a mixture with the aid of ultrasound for 20 minutes, filter through
suitable quantity of methanol to obtain a mixture containing a membrane filter disc with an average pore diameter not greater
0.1 per cent w/v of Efavirenz and filter through a membrane than 1.0 µm, rejecting the first few ml of the filtrate, and dilute
filter disc with an average pore diameter not exceeding 1.0 µm, 10.0 ml of the filtrate to 50.0 ml with methanol.
rejecting the first few ml of the filtrate. Reference solution. A 0.12 per cent w/v solution of efavirenz
Reference solution. A 0.1 per cent w/v solution of efavirenz RS in methanol.
RS in methanol. Chromatographic system
Chromatographic system – a stainless steel column 25 cm x 4.6 mm, packed with
– a stainless steel column 25 cm x 4.6 mm, packed with base deactivated octadecylsilyl silica gel (5 µm),
base deactivated octadecylsilyl silica gel (5 µm), – mobile phase: a filtered and degassed mixture of
– mobile phase: a filtered and degassed mixture of 50 50 volumes of acetonitrile and 50 volumes of a 0.86 per
volumes of acetonitrile and 50 volumes of a 0.86 per cent w/v solution of ammonium dihydrogen phosphate,
cent w/v solution of ammonium dihydrogen phosphate, the pH of which is adjusted to 3.0 ± 0.05 with phosphoric
the pH of which is adjusted to 3.0 ± 0.05 with phosphoric acid,
acid, – flow rate. 1.5 ml per minute,
– flow rate. 1.5 ml per minute, – spectrophotometer set at 252 nm,
– a 20 µl loop injector. Inject the reference solution. The test is not valid unless the
Inject the reference solution. The test is not valid unless the column efficiency determined from the efavirenz peak is not
column efficiency determined from the efavirenz peak is not less than 6000 theoretical plates and the tailing factor is not
less than 6000 theoretical plates and the tailing factor is not more than 2.0 and the relative standard deviation is not more
more than 2.0. than 2.0 per cent.
455
EMETINE HYDROCHLORIDE IP 2007
Inject alternately the test solution and the reference solution. pH (2.4.24). 4.0 to 6.0, determined in a 2.0 per cent w/v solution.
Calculate the content of C14H9ClF3NO2. Specific optical rotation (2.4.22). +16.0 ° to +19.0°, determined
Emetine Hydrochloride
volumes of 2-methoxyethanol, 10 volumes of methanol and 1
H3CO volume of diethylamine.
N
H3CO Solvent mixture. A 1 per cent v/v solution of 2 M ammonia in
H
CH3 methanol.
,2HCl,7H2O
H
OCH3 examination in 100 ml with solvent mixture.
HN
Reference solution (a). A 0.05 per cent w/v solution of emetine
OCH3 hydrochloride RS in the same solvent.
C29H40N2O4,2HCl,7H2O Mol. Wt. 679.7 isoemetine hydrobromide RS in the same solvent.
Emetine Hydrochloride is 6',7',10,11-tetramethoxyemetan Reference solution (c). A 0.001 per cent w/v solution of
dihydrochloride heptahydrate. cephaeline hydrochloride RS in the same solvent.
Emetine Hydrochloride contains not less than 98.0 per cent Reference solution (d). A 0.0005 per cent w/v solution of
and not more than 102.0 per cent of C29H40N2O4,2HCl, emetine hydrochloride RS in the same solvent.
calculated on the dried basis. Reference solution (e). Mix 1 ml each of reference solutions
Description. A white or very slightly yellowish, crystalline (a), (b) and (c). Prepare immediately before use.
powder; odourless. Develops a faint yellow tint on exposure Apply to the plate 10 µl of each of the test solution and
to light. reference solutions (a) to (d) and 30 µl of reference solution
(e). After development, dry the plate in air until the odour of
Identification solvent is no longer detectable, spray with a 0.5 per cent w/v
Test A may be omitted if tests B, C and D are carried out. Tests solution of iodine in chloroform, heat at 60° for 15 minutes
B and C may be omitted if tests A and D are carried out. and examine in ultraviolet light at 365 nm. Any spots
corresponding to isoemetine and cephaeline in the
A. Determine by infrared absorption spectrophotometry (2.4.6). chromatogram obtained with the test solution are not more
Compare the spectrum with that obtained with emetine intense than the spots in the chromatograms obtained with
hydrochloride RS or with the reference spectrum of emetine reference solution (b) and (c) respectively. Any other
hydrochloride. secondary spot in the chromatogram obtained with the test
B. In the test for Related substances, the principal spot in the solution is not more intense than the spot in the chromatogram
chromatogram obtained with the test solution corresponds to obtained with reference solution (d). The test is not valid
that in the chromatogram obtained with the reference solution unless the chromatogram obtained with reference solution (e)
(a). shows three clearly separated spots.
C. Sprinkle 5 mg on the surface of 1 ml of a 5 per cent w/v Sulphated ash (2.3.18). Not more than 0.1 per cent.
solution of ammonium molybdate in sulphuric acid; a bright Loss on drying (2.4.19). 15.0 to 19.0 per cent, determined on
green colour develops. 0.5 g by drying in an oven at 105° for 3 hours.
D. Gives reaction A of chlorides (2.3.1). Assay. Weigh accurately about 0.4 g, dissolve in 40 ml of
anhydrous glacial acetic acid and add 7 ml of mercury acetate
Tests solution. Titrate with 0.1 M perchloric acid, using crystal
Appearance of solution. A 5.0 per cent w/v solution is clear violet solution as indicator. Carry out a blank titration.
(2.4.1) and not more intensely coloured than reference solution 1 ml of 0.1 M perchloric acid is equivalent to 0.02768 g of
YS5 or BYS5 (2.4.1). C29H40N2O4,2HCl.
456
IP 2007 EMTRICITABINE
Storage. Store protected from light. Emtricitabine

NH2
F
Emetine Injection N
Emetine Hydrochloride Injection O N

O
Emetine Injection is a sterile solution of Emetine Hydrochloride
in Water for Injections. HO
S
Emetine Injection contains not less than 95.0 per cent and not
more than 105.0 per cent of the stated amount of emetine C8H10FN3O3S Mol. Wt. 247.3
hydrochloride, C29H40N2O4,2HCl,7H2O. Emtricitabine is 4-amino-5-fluoro-1-[(2R,5S)-2-
(hydroxymethyl)-1,3-oxathiolan-5-yl]-2-(1H)-pyrimidone.
Identification
Emtricitabine contain not less than 98.0 per cent and not more
A. To a volume of the injection containing 0.2 g of Emetine than 102.0 per cent of C8H10FN3O3S, calculated on the dried
Hydrochloride add an excess of 5 M sodium hydroxide and basis.
extract with ether. Evaporate the ether, dissolve the residue in Description. A white to off-white, crystalIine powder.
a few ml of ethanol (95 per cent), add 0.25 ml of carbon
disulphide, boil and scratch the sides of the container with a Identification
glass rod; a crystalline precipitate separates which melts at
about 206° (2.4.21). A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with emtricitabine
B. Evaporate 1 ml on a water-bath to dryness. The residue RS or with the reference spectrum of emtricitabine.
complies with the following tests.
Sprinkle 5 mg on the surface of 1 ml of a 5 per cent w/v solution obtained with the test solution corresponds to the peak in the
of ammonium molybdate in sulphuric acid; a bright green chromatogram obtained with the reference solution.
colour develops.
C. Gives reaction A of chlorides (2.3.1). Tests
Specific optical rotation (2.4.22). - 125.0º to - 150.0º, determined
Tests in a 0.5 per cent w/v solution in methanol.
pH (2.4.24). 2.7 to 4.0. Enantiomeric purity. Determine by liquid chromatography
Other tests. Complies with the tests stated under Parenteral (2.4.14).
Preparations (Injections). Test solution. Dissolve 25 mg of the substance under
Assay. Dilute a volume containing about 0.2 g of Emetine examination in 50 ml of the mobile phase.
Hydrochloride to 20 ml with water, add 10 ml of 5 M sodium Reference solution. Dissolve 25 mg of racemic emtricitabine
hydroxide and shake with successive quantities, each of RS in 25 ml of the mobile phase.
50 ml, of ether until complete extraction of the alkaloid is
effected. Wash the combined ether extracts with successive
– a stainless steel column 25 cm x 4.6 mm, packed with a
quantities, each of 10 ml, of water until the washings, after
chiral stationary phase (5 µm) (such as Chirobiotic V),
extraction with a further 50 ml of ether, are neutral to litmus
– mobile phase: a mixture of 1000 volumes of methanol, 2
paper. Mix the ether solutions, add 20 ml of water and 10.0 ml
volumes of diethyl amine and 1volume of glacial acetic
of 0.1 M hydrochloric acid, shake, allow to separate and
acid,
collect the aqueous layer, shake the ether solution with two
further quantities, each of 20 ml, of water, mix the aqueous
solutions and titrate with 0.1 M sodium hydroxide using
methyl red solution as indicator.
Inject the reference solution. The elution order is, the 5-fluoro-
1 ml of 0.1 M hydrochloric acid is equivalent to 0.03398 g of
1-(2R,5S)-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine
C29H40N2O4,2HCl,7H2O.
isomer followed by the other isomer. The resolution between
Storage. Store protected from light. the two isomers should not be less than 2.0.
457
EMTRICITABINE CAPSULES IP 2007
Inject the test solution and measure the areas of the two – mobile phase: a mixture of 95 volumes of 0.025 M
isomers. ammonium acetate solution adjusted the pH to 5.0 and
Calculate the content of the 5-fluoro-1-(2R,5S)-[2- 5 volumes of methanol,
(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine isomer by area – flow rate. 1 ml per minute,
normalization, not less than 99.0 per cent. – spectrophotometer set at 277 nm,
(2.4.14). Inject the reference solution. The test is not valid unless the
Test solution. Dissolve 25 mg of the substance under less than 2000 theoretical plates and the relative standard
examination in 25 ml of the mobile phase. deviation for replicate injections is not more than 2.0 per cent.
emtricitabine RS in the mobile phase.
Reference solution (b). Dilute 1 ml of reference solution (a) to Calculate the content of C8H10FN3O3S.
100 ml with the mobile phase. Storage. Store protected from light and moisture.
pentaflurophenyl bonded to silica (5 µm) (such as F-5
Supelco discovery) Emtricitabine Capsules
– mobile phase: a mixture of 99 volumes of 0.025 M Emtricitabine Capsules contain not less than 90.0 per cent and
ammonium acetate solution with the pH adjusted to 5.0 not more than 110.0 per cent of the stated amount of
with glacial acetic acid, and 1 volume of methanol, emtricitabine, C8H10FN3O3S.
Inject reference solution (a). The test is not valid unless the obtained with the test solution corresponds to the peak in the
tailing factor is not more than 2.0, the column efficiency in not chromatogram obtained with the reference solution.
less than 2000 theoretical plates.
B. Disperse the quantity of contents of the capsules containing
Inject the test solution and the reference solution (b). In the 1 mg of Emtricitabine in 100 ml with methanol and filter.
secondary peak is not more than 0.5 times the area of the peak When examined in the range 200 nm to 400 nm (2.4.7), the
due to the reference solution (b) (0.5 per cent) and the sum of filtrate shows absorption maxima at the same wavelengths as
areas of all the secondary peaks is not more than twice the 0.001 per cent w/v solution of emtricitabine RS in methanol.
area of the peak due to the reference solution (b) (2.0 per cent).
Tests
Heavy metals (2.3.13). 1 g complies with the limit test for heavy
metals, Method B (20 ppm). Dissolution (2.5.2).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Apparatus. No 1
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Medium. 900 ml of 0.1 M hydrochloric acid.
on 1 g by drying in an oven at 105º. Speed and time. 50 rpm and 30 minutes.
Assay. Determine by liquid chromatography (2.4.14). Withdraw a suitable volume of the medium and filter promptly
not more than 0.5 µm, rejecting the first 2 ml of the filtrate.
examination in 25.0 ml of the mobile phase. Dilute 5.0 ml of the
solution to 50.0 ml with the mobile phase. Determine by liquid chromatography (2.4.14).
Reference solution. A 0.1 per cent w/v solution of Test solution. Use the filtrate.
emtricitabine RS in the mobile phase. Dilute 5.0 ml of the
Reference solution. Dissolve 27.5 mg of emtricitabine RS in
solution to 50.0 ml with the mobile phase.
15 ml of methanol, dilute to 25 ml with the mobile phase. Dilute
Chromatographic system 2 ml of the solution to 10 ml with the dissolution medium.
Use the chromatographic system described under Assay.
Supelco discovery), Inject the test solution and the reference solution.
458
IP 2007 EMULSIFYING WAX
D. Not less than 75 per cent of the stated amount of 1000 ml of water and adjusting the pH to 3.8 with glacial
C8H10FN3O3S. acetic acid, and 20 volumes of methanol,
Related substances. Determine by liquid chromatography – flow rate. 1 ml per minute,
(2.4.14). – spectrophotometer set at 277 nm,
Test solution. Weigh a quantity of the contents of the capsules Inject the reference solution. The test is not valid unless the
containing 100 mg of Emtricitabine, disperse in 100 ml of the tailing factor is not more than 2.0, the column efficiency in not
mobile phase and filter. less than 2000 theoretical plates and the relative standard
Reference solution (a). A 0.1 per cent w/v solution of deviation for replicate injections is not more than 2.0 per cent.
emtricitabine RS in the mobile phase. Inject the test solution and the reference solution.
Reference solution (b). Dilute 1 ml of reference solution (a) to Calculate the content of C8H10FN3O3S in the capsules.
Supelco discovery), Emulsifying Wax
Anionic Emulsifying Wax
prepared by dissolving 1.9 g of ammonium acetate in
1000 ml of water and adjusting the pH to 5.0 with glacial Emulsifying Wax is a waxy solid containing 90 parts of
acetic acid, and 1 volume of methanol, Cetostearyl Alcohol, 10 parts of Sodium Lauryl Sulphate or
– flow rate. 1.5 ml per minute, sodium salts of similar sulphated higher primary aliphatic
– spectrophotometer set at 277 nm, alcohols, and 4 parts of Purified Water.
– a 10 µl loop injector. Description. An almost white or pale yellow, waxy solid or
Inject reference solution (a). The test is not valid unless the flakes; odour, faint and characteristic. It becomes soft on
tailing factor is not more than 2.0 and the column efficiency in warming.
not less than 2000 theoretical plates.
Inject the test solution and reference solution (b). In the Identification
chromatogram obtained with the test solution, the area of any The residue obtained in the test for Unsaponifiable matter
secondary peak is not more than the area of the peak due to melts at about 52° (2.4.21).
the reference solution (b) (1.0 per cent) and the sum of areas
of all the secondary peaks is not more than 3 times the area of Tests
the peak due to the reference solution (b) (3.0 per cent).
Acidity. Weigh accurately about 20.0 g, add a mixture of 40 ml
of ether and 75 ml of ethanol (95 per cent), previously
Water (2.3.43). Not more than 5.0 per cent, determined on neutralised to phenolphthalein solution, and warm gently
0.5 g. until solution is effected. Titrate with 0.1 M sodium hydroxide
Assay. Determine by liquid chromatography (2.4.14). using phenolphthalein solution as indicator until a pink colour
which persists for at least 15 seconds is obtained. Not more
Test solution. Weigh accurately a quantity of the mixed than 1.0 ml of 0.1 M sodium hydroxide is required.
contents of 20 capsules containing about 100 mg of
Emtricitabine, disperse in 100.0 ml of methanol and filter. Dilute Alkalinity. 25 ml of a 20 per cent w/v dispersion in warm
5.0 ml of the solution to 50.0 ml with the mobile phase. ethanol (95 per cent), previously neutralised to
phenolphthalein solution and cooled, exhibits no colour on
Reference solution. A 0.1 percent w/v solution of emtricitabine the addition of 0.5 ml of phenolphthalein solution.
RS in methanol. Dilute 5.0 ml of the solution to 50.0 ml with the
mobile phase. Saponification value (2.3.37). Not more than 2.0, determined
on 20.0 g.
– a stainless steel column 25 cm x 4.6 mm, packed with Unsaponifiable matter (2.3.39). Not less than 86.0 per cent,
octadecylsilane bonded to porous silica (5 µm) (such as calculated on the anhydrous basis, determined on 5 g and
Intersil ODS 3V), omitting the titration of the residue.
– mobile phase: a mixture of 80 volumes of a buffer solution Iodine value (2.3.28). Not more than 3.0, determined by the
prepared by dissolving 1.9 g of ammonium acetate in iodine monochloride method.
459
ENALAPRIL MALEATE IP 2007
Alcohols. To 3.5 g of the residue obtained in the test for Heavy metals (2.3.13). 2.0 g complies with the limit test for
Unsaponifiable matter add 12 g of stearic anhydride and heavy metals, Method B (10 ppm).
10 ml of xylene and heat gently under a reflux condenser for Sulphated ash (2.3.18). Not more than 0.2 per cent.
30 minutes. Cool, add a mixture of 40 ml of pyridine and 4 ml of
water, reflux for a further 30 minutes and titrate the hot solution Loss on drying (2.4.19). Not more than 1.0 per cent, determined
with 1 M sodium hydroxide using phenolphthalein solution on 1.0 g by drying in an oven at 60° at a pressure not exceeding
as indicator. Repeat the operation omitting the residue. The 0.7 kPa for 2 hours.
difference between the titrations is not less than 12.8 ml and Assay. Determine by liquid chromatography (2.4.14).
not more than 14.2 ml.
Test solution. Weigh accurately about 30 mg of the substance
Sodium alkyl sulphates. Not less than 8.7 per cent, calculated under examination and dissolve in 100.0 ml of the mobile phase.
as C12H25O4SNa, on the anhydrous basis, determined by the
following method. Weigh accurately about 0.25 g, dissolve as Reference solution. A freshly prepared 0.03 per cent w/v
completely as possible in 15 ml of chloroform, add 30 ml of solution of enalapril maleate RS in the mobile phase.
water, 10 ml of 1 M sulphuric acid and 1 ml of dimethyl yellow- Chromatographic system
oracet blue B solution and titrate with 0.004 M benzethonium – a stainless steel column 25 cm x 4.6 mm, packed with a
chloride, shaking vigorously and allowing the layers to rigid spherical styrene divinylbenzene copolymer (5 to
separate after each addition, until the chloroform layer acquires 10 µm),
a permanent clear green colour. – mobile phase: a mixture of 4 volumes of mixed phosphate
1 ml of 0.004 M benzethonium chloride is equivalent to buffer pH 6.8 and 1 volume of acetonitrile,
0.001154 g of C12H25O4SNa. – column temperature 70°,
Water (2.3.43). Not more than 4.0 per cent, determined on 0.6 g. – spectrophotometer set at 210 nm,
Inject the references solution. The test is not valid unless the
Enalapril Maleate column efficiency is not less than 300 theoretical plates and
the relative standard deviation for the area of the peak due to
O COOH enalapril maleate is not more than 1.0 per cent.
H
N COOH
N , Inject the test solution and reference solution.
CH3 COOH
H3C O O Calculate the content of C20H28N2O5,C4H4O4.
C20H28N2O5,C4H4O4 Mol. Wt. 492.5

Enalapril Maleate is (2S)-1-[(2S)-2-[[(1S)-1-(ethoxycarbonyl)- Enalapril Maleate Tablets
3-phenylpropyl]amino]propanoyl]pyrrolidin-2-carboxylic
acid hydrogen maleate. Enalapril Maleate Tablets contain not less than 90.0 per cent
Enalapril Maleate contains not less 98.0 per cent and not more enalapril maleate, C20H28N2O5,C4H4O4.
than 102.0 per cent of C20H28N2O5,C4H4O4, calculated on the
dried basis. Identification
Description. An off-white, crystalline powder. In the Assay, the principal peak in the chromatogram obtained
Identification chromatogram obtained with the reference solution.
Compare the spectrum with that obtained with enalapril
Tests
maleate RS or with the reference spectrum of enalapril maleate. Uniformity of content (For tablets containing 10 mg or less).
B. Melts at about 144° (2.4.21). Comply with the test stated under Tablets.
Tests
Test solution. Finely crush one tablet, transfer to a 50-ml
Specific optical rotation (2.4.22). –41.0° to –43.5°, determined volumetric flask, add about 30 ml of mixed phosphate buffer
in a 1.0 per cent w/v solution in methanol. pH 2.0, disperse with the aid of ultrasound for 15 minutes,
460
IP 2007 EPHEDRINE
shake for another 30 minutes, dilute to volume with the buffer Description. Colourless crystals or a white, crystalline powder.
solution, mix and filter. Dilute a portion of the filtrate with the Gradually decomposes on exposure to light.
buffer solution to obtain a solution containing 0.01 per cent
w/v of Enalapril Maleate. Identification
Reference solution. A 0.01 per cent w/v solution of enalapril Test A may be omitted if tests B, C, D and E are carried out.
maleate RS in the same buffer solution. Tests B and C may be omitted if tests A, D and E are carried
Chromatographic system out.
– a stainless steel column 20 cm x 4.6 mm, packed with A. Determine by infrared absorption spectrophotometry (2.4.6)
octylsilyl silica gel (3 to 10 µm), on discs prepared in the following manner. Separately dissolve
– mobile phase: a filtered and degassed mixture of 40 mg of the substance under examination and 50 mg of
75 volumes of phosphate buffer pH 2.0 and 25 volumes ephedrine hydrochloride RS in 1 ml of water, add 1 ml of 2 M
of acetonitrile, sodium hydroxide and 4 ml of chloroform, shake and dry the
– column. temperature 50°, organic layer with 0.2 g of anhydrous sodium sulphate. Prepare
– flow rate. 2 ml per minute, the discs using 0.3 g of potassium bromide IR, apply dropwise
– spectrophotometer set at 215 nm, to the discs 0.1 ml of the chloroform layer, allowing the solvent
– a 50 µl loop injector. to evaporate between applications, and dry the discs at 50°
Inject alternately the test solution and the reference solution. for 2 minutes.
Calculate the content of C20H28N2O5,C4H4O4 in the tablet. Compare the spectrum with that obtained with the base isolated
from ephedrine hydrochloride RS or with the reference
Other tests. Complies with the tests stated under Tablets. spectrum of ephedrine.
Assay. Determine by liquid chromatography (2.4.14). B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.05
Test solution. Weigh and powder 20 tablets. Weigh accurately per cent w/v solution in 0.1 M hydrochloric acid shows
a quantity of the powder containing about 50 mg of Enalapril absorption maxima at about 251 nm, 259 nm and 265 nm;
Maleate, add 150 ml of phosphate buffer pH 2.0, disperse with absorbance at about 251 nm, about 0.37; at about 259 nm,
the aid of ultrasound for 15 minutes, shake for another about 0.48; at about 265 nm about 0.36.
30 minutes and dilute with the buffer to 250.0 ml, mix and filter. C. In the test for Related substances, the principal spot in the
Reference solution. A 0.02 per cent w/v solution of enalapril chromatogram obtained with test solution (b) corresponds to
maleate RS in phosphate buffer pH 2.0. that in the chromatogram obtained with reference solution
(b).
Follow the chromatographic procedure described under
Uniformity of content. D. Dissolve 10 mg in 1 ml of water, add 0.2 ml of dilute
hydrochloric acid and add 0.1 ml of cupric sulphate solution
Calculate the content of C20H28N2O5,C4H4O4 in the tablets. followed by 1 ml of sodium hydroxide solution; a violet colour
is produced. Add 2 ml of ether and shake; the ether layer is
purple and the aqueous layer is blue.
Ephedrine E. Melting range (2.4.21). 40° to 43° (hydrated material),
determined on the undried substance. The anhydrous material
OH H melts at about 36°.
N
CH3 Tests
CH3 Appearance of solution. A 2.5 per cent w/v solution is clear
C10H15NO Mol. Wt. 165.2 (anhydrous)
Specific optical rotation (2.4.22). –41.0° to –43.0°, determined
C10H15NO, ½ H2O Mol. Wt. 174.2 (hemihydrate) at 20° in a solution prepared by dissolving 2.25 g in 15 ml of
Ephedrine is (1R,2S)-2-methylamino-1-phenylpropan-1-ol, dilute hydrochloric acid and diluting to 50.0 ml with water.
an alkaloid obtained from Ephedra or prepared Related substances. Determine by thin-layer chromatography
synthetically. It may be anhydrous or a hemihydrate. (2.4.17), coating the plate with silica gel G.
Ephedrine contains not less than 98.5 per cent and not more Mobile phase. A mixture of 80 volumes of 2-propanol,15
than 101.0 per cent of C10H15NO, calculated on the anhydrous volumes of strong ammonia solution and 5 volumes of
basis. chloroform.
461
EPHEDRINE HYDROCHLORIDE IP 2007
Test solution (a). Dissolve 0.2 g of the substance under A. Determine by infrared absorption spectrophotometry (2.4.6).
examination in 10 ml of methanol. Compare the spectrum with that obtained with ephedrine
Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with hydrochloride RS or with the reference spectrum of ephedrine
methanol. hydrochloride.
Reference solution (a). Dilute 5 ml of test solution (b) to 100 B. In the test for Related substances, the principal spot in the
ml with methanol. chromatogram obtained with test solution (b) corresponds to
ephedrine hydrochloride RS in methanol. C. Dissolve 10 mg in 1 ml of water and add 0.1 ml of cupric
sulphate solution and 1 ml of sodium hydroxide solution; a
Apply to the plate 10 µl of each solution. After development, violet colour is produced. Add 2 ml of ether and shake; the
dry the plate in air, spray with ninhydrin solution and heat at ether layer is purple and the aqueous layer is blue.
110° for 5 minutes. Any secondary spot in the chromatogram
obtained with test solution (a) is not more intense than the D. A 5 per cent w/v solution gives reaction A of chlorides
spot in the chromatogram obtained with reference solution (2.3.1).
(a). Ignore any spot of lighter colour than the background.
Tests
Chlorides. Dissolve 0.18 g in 10 ml of water, add 5 ml of 2 M
nitric acid and 0.5 ml of silver nitrate solution and allow to Appearance of solution. A 10.0 per cent w/v solution (Solution
stand for 2 minutes protected from bright light. Any A) is clear (2.4.1), and colourless (2.4.1).
opalescence produced is not more intense than that obtained Acidity or alkalinity. To 10 ml of Solution A add 0.1 ml of
by repeating the operation at the same time and in the same methyl red solution and 0.2 ml of 0.01 M sodium hydroxide;
manner using 2.0 ml of chloride standard solution (25 ppm the solution is yellow. Add 0.4 ml of 0.01 M hydrochloric
Cl) in place of the solution of the substance under examination acid; the solution is red.
(280 ppm). Specific optical rotation (2.4.22). –33.5° to –35.5°, determined
Sulphated ash (2.3.18). Not more than 0.1 per cent. in a 5.0 per cent w/v solution.
Water (2.3.43). 4.5 to 5.5 per cent (for hemihydrate) and not Related substances. Determine by thin-layer chromatography
more than 1.0 per cent (for anhydrous), determined on 0.3 g. (2.4.17), coating the plate with silica gel G.
Assay. Weigh accurately about 0.5 g and dissolve in 5 ml of Mobile phase. A mixture of 80 volumes of 2-propanol,15
ethanol (95 per cent). Add 50.0 ml of 0.1 M hydrochloric volumes of strong ammonia solution and 5 volumes of
acid and titrate with 0.1 M sodium hydroxide using methyl chloroform.
red solution as indicator until a yellow colour is obtained. Test solution (a). Dissolve 0.2 g of the substance under
1 ml of 0.1 M hydrochloric acid is equivalent to 0.01652 g of examination in 10 ml of methanol.
C10H15NO. Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with
Storage. Store protected from light. methanol.
Reference solution (a). Dilute 5 ml of test solution (b) to100
ml with methanol.
Ephedrine Hydrochloride Reference solution (b). A 0.2 per cent w/v solution of
C10H15NO,HCl Mol. Wt. 201.7 ephedrine hydrochloride RS in methanol.
Ephedrine Hydrochloride is (1R,2S)-2-methylamino-1- Apply to the plate 10 µl of each solution. After development,
phenylpropan-1-ol hydrochloride. dry the plate in air, spray with ninhydrin solution and heat at
Ephedrine Hydrochloride contains not less than 99.0 per cent
and not more than 101.0 per cent of C10H15NO,HCl calculated
on the dried basis.
Description. Colourless crystals or a white, crystalline powder;
Sulphates (2.3.17). 15 ml of solution A complies with the limit
odourless. It is affected by light.
test for sulphates (100 ppm).
Test A may be omitted if tests B, C and D are carried out. Tests Loss on drying (2.4.19). Not more than 0.5 per cent, determined
B and C may be omitted if tests A and D are carried out. on 1.0 g by drying in an oven at 105°.
462
IP 2007 EPHEDRINE ORAL SOLUTION
Assay. Weigh accurately about 0.17 g, dissolve in 10 ml of Reference solution (b). A 5.0 per cent v/v solution of the
mercuric acetate solution, warming gently, add 50 ml of internal standard in water.
acetone and mix. Titrate with 0.1 M perchloric acid, using 1 Chromatographic system
ml of a saturated solution of methyl orange in acetone as – a column 1.5 m x 4 mm, packed with porous polymer
indicator, until a red colour is obtained. Carry out a blank beads (100 to 200 mesh) (such as Porapak Q and
titration. Chromosorb 101),
1 ml of 0.1 M perchloric acid is equivalent to 0.02017 g of – temperature:
C10H15NO,HCl. column.150°,
Calculate the percentage content of ethanol from the areas of
the peaks due to ethanol in the chromatograms obtained with
Ephedrine Oral Solution reference solutions (a) and (b).
Ephedrine Hydrochloride Oral Solution; Ephedrine Related substances. Determine by thin-layer chromatography
Hydrochloride Elixir; Ephedrine Elixir (2.4.17), coating the plate with silica gel G.
Ephedrine Oral Solution is a solution containing 0.3 per cent Mobile phase. A mixture of 80 volumes of 2-propanol,15
w/v of Ephedrine Hydrochloride in a suitable flavoured vehicle volumes of strong ammonia solution and 5 volumes of
containing a sufficient volume of Ethanol (95 per cent) or of chloroform.
an appropriate dilute ethanol to give a final concentration of Test solution (a). Add sufficient 5 M ammonia to 50 ml of the
not more than 3 per cent v/v of ethanol. oral solution to make it alkaline, extract with two quantities,
Ephedrine Oral Solution contains not less than 0.27 per cent each of 100 ml, of ether, wash the combined extracts with 10 ml
and not more than 0.33 per cent w/v of ephedrine of water, dry with anhydrous sodium sulphate, filter and
hydrochloride, C10H15NO,HCl. evaporate the filtrate to dryness. Dissolve the oily residue in
sufficient methanol to produce 5 ml.
Identification Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with
A. To 30 ml add 2 ml of 2 M hydrochloric acid, extract with methanol.
two quantities, each of 20 ml, of ether and discard the ether. Reference solution (a). Dilute 1 ml of test solution (a) to 200
Add sufficient dilute ammonia solution to the aqueous phase ml with methanol.
to make it alkaline, extract with two quantities, each of 30 ml, of
ether, wash the combined ether extracts with three quantities,
ephedrine hydrochloride RS in methanol.
each of 15 ml, of water, dry over anhydrous sodium sulphate,
filter and evaporate the filtrate to dryness. The residue complies Apply to the plate 10 µl of each solution. After development,
with the following test. dry the plate in air, spray with ninhydrin solution and heat at
Compare the spectrum with that obtained with ephedrine
reference spectrum of ephedrine.
Other tests. Complies with the tests stated under Oral Liquids.
chromatogram obtained with test solution (b) corresponds to Assay. Determine by liquid chromatography (2.4.14).
that in the chromatogram obtained with reference solution (b). Test solution. Dilute an accurately weighed quantity of the
oral solution containing 60 mg of Ephedrine Hydrochloride to
Tests
Ethanol content. Not more than 3 per cent v/v, determined by Reference solution. A 0.12 per cent w/v solution of ephedrine
gas chromatography (2.4.13). hydrochloride RS in methanol.
Test solution. Use the preparation under examination. Chromatographic system
Reference solution (a). Add sufficient of 1-propanol (internal – a stainless steel column 20 cm x 4.6 mm, packed with
standard) to the test solution to produce a solution containing octadecylsilyl silica gel (5 µm),
5.0 per cent v/v of 1-propanol. – mobile phase: 0.005 M dioctyl sodium sulphosuccinate
463
EPHEDRINE TABLETS IP 2007
in a mixture of 65 volumes of methanol, 35 volumes of Tests

water and 1 volume of glacial acetic acid,
– a 20 µl loop injector. Mobile phase. A mixture of 80 volumes of 2-propanol,15
volumes of strong ammonia solution and 5 volumes of
Determine the weight per ml of the oral solution (2.4.29), and
chloroform.
calculate the content of C10H15NO,HCl, weight in volume.
Test solution (a). Extract a quantity of the powdered tablets
containing 0.1 g of Ephedrine Hydrochloride with 5 ml of
methanol and filter.
Ephedrine Tablets Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with
methanol.
Ephedrine Hydrochloride Tablets Reference solution (a). Dilute 1 ml of test solution (a) to
Ephedrine Tablets contain not less than 92.5 per cent and not 200 ml with methanol.
more than 107.5 per cent of the stated amount of ephedrine Reference solution (b). A 0.2 per cent w/v solution of
hydrochloride, C10H15NO,HCl. ephedrine hydrochloride RS in methanol.
Identification Apply to the plate 10 µl of each solution. After development,
dry the plate in air, spray with ninhydrin solution and heat at
A. Shake a quantity of the powdered tablets containing 0.1 g 110° for 5 minutes. Any secondary spot in the chromatogram
of Ephedrine Hydrochloride with 20 ml of 0.1 M hydrochloric obtained with test solution (a) is not more intense than the
acid, filter, wash the filtrate with two quantities, each of 20 ml, spot in the chromatogram obtained with reference solution
of chloroform and discard the chloroform. Make the aqueous (a). Ignore any spot of lighter colour than the background.
layer alkaline with 5 M ammonia and extract with two quantities, Other tests. Comply with the tests stated under Tablets.
each of 30 ml, of a mixture of 3 volumes of chloroform and 1
Assay. Weigh 20 tablets and reduce to a fine powder. Weigh
volume of ethanol (95 per cent). Dry the combined extracts
accurately a quantity of the powder containing about 0.15 g
over anhydrous sodium sulphate, filter and evaporate to a
of Ephedrine Hydrochloride and add 30 ml of anhydrous
low volume at a pressure of 2 kPa. Prepare a disc using 0.3 g of
glacial acetic acid and 10 ml of mercuric acetate solution.
potassium bromide IR, apply dropwise to the disc 0.1 ml of
Warm gently to effect solution and cool. Titrate with 0.1 M
the chloroform solution, allowing the solvent to evaporate
perchloric acid, using 0.1 ml of crystal violet solution as
between applications, and dry the disc at 50° for 2 minutes.
indicator, until the violet colour changes to green-blue. Carry
The disc so obtained complies with the following test.
Compare the spectrum with that obtained with ephedrine
C10H15NO,HCl.
reference spectrum of ephedrine. Storage. Store protected from light.
chromatogram obtained with test solution (b) corresponds to Ergocalciferol
Calciferol; Vitamin D2
C. Triturate a quantity of the powdered tablets containing
about 0.4 g of Ephedrine Hydrochloride with 10 ml of
CH3
chloroform and discard the chloroform. Repeat trituration with
H 3C CH3
a further 10 ml of chloroform and again discard the chloroform.
Shake the residue with 30 ml of warm ethanol (95 per cent) for H3C
CH3
20 minutes, filter, evaporate the filtrate to dryness on a water-
H2C
bath and dry the residue at 80° (residue R). Dissolve 10 mg of
residue R in 1 ml of water and add 0.1 ml of cupric sulphate H
solution followed by 1 ml of sodium hydroxide solution; a
HO
violet colour is produced. Add 1 ml of ether and shake; the
ether layer is purple and the aqueous layer is blue. C28H44O Mol. Wt. 396.7
D. A 5 per cent w/v solution of residue R gives reaction A of Ergocalciferol is (5Z,7E,22E)-(3S)-9,10-secoergosta-
chlorides (2.3.1). 5,7,10(19),22-tetraen-3-ol.
464
IP 2007 ERGOCALCIFEROL
Ergocalciferol contains not less than 97.0 per cent and not per cent) and beginning at the words “add 0.5 ml of a 0.5 per
more than 103.0 per cent of C28H44O. cent w/v solution....”.
Description. White or almost white crystals or a white or Ergosterol. Determine by thin-layer chromatography (2.4.17),
slightly yellowish, crystalline powder. It is sensitive to air, coating the plate with silica gel G.
heat and light. A reversible isomerisation to pre-ergocalciferol Mobile phase. A 0.01 per cent w/v solution of butylated
may occur in solution, depending on temperature and time. hydroxytoluene in a mixture of equal volumes of cyclohexane
and peroxide-free ether.
Identification
Test A may be omitted if tests B, C and D are carried out. Tests examination in sufficient 1,2-dichloroethane containing 1 per
B, C and D may be omitted if test A is carried out. cent w/v of squalane and 0.1 per cent w/v butylated
A. Determine by infrared absorption spectrophotometry (2.4.6). hydroxytoluene (solvent A) to produce 5 ml.
Compare the spectrum with that obtained with ergocalciferol Reference solution (a). A 5.0 per cent w/v solution of
RS. ergocalciferol RS in solvent A.
B. Dissolve 1 mg in 1 ml of 1,2-dichloroethane and 4 ml of Reference solution (b). A 0.01 per cent w/v solution of
antimony trichloride solution; a yellowish-orange colour is ergosterol RS in solvent A.
produced.
Reference solution (c). Mix equal volumes of reference
C. In the test for Ergosterol, the principal spot in the solutions (a) and (b).
Apply to the plate 10 µl of each solution. Develop the
that in the chromatogram obtained with the reference solution
chromatograms immediately, protected from light. After
(a).
development dry the plate in air and spray three times with
D. To a solution of about 0.5 mg in 5 ml of chloroform add antimony trichloride reagent. Examine the chromatograms
0.3 ml of acetic anhydride and 0.1 ml of sulphuric acid and for not more than 4 minutes after spraying. The principal spot
shake vigorously; a bright red colour is produced which rapidly in the chromatogram obtained with the test solution is initially
changes through violet and blue to green. orange-yellow but becomes brown later. In the chromatogram
obtained with the test solution any violet spot with an
Tests
Rf value slightly lower than that of the principal spot (due to
Specific optical rotation (2.3.22). +103° to +107°, determined ergosterol and appearing slowly) is not more intense than the
within 30 minutes of preparation, in a solution prepared by spot in the chromatogram obtained with reference solution
dissolving 0.2 g rapidly and without heating in sufficient (b). The chromatogram obtained with the test solution shows
aldehyde-free ethanol (95 per cent) to produce 25.0 ml. no spot that does not correspond to one of the spots in the
Light absorption. Dissolve 10 mg rapidly and without heating chromatograms obtained with reference solutions (a) and (b).
in sufficient aldehyde-free ethanol (95 per cent) to produce The test is not valid unless the chromatogram obtained with
100.0 ml. Dilute 5.0 ml of this solution to 50.0 ml with aldehyde- reference solution (c) shows two clearly separated principal
free ethanol (95 per cent). Absorbance of the resulting spots.
solution at the maximum at about 265 nm (2.4.7), measured Assay. Carry out the following procedure as rapidly as
within 30 minutes of preparation, 0.45 to 0.50. possible in subdued light and protected from air.
Reducing substances. To 10 ml of a 1 per cent w/v solution in Determine by liquid chromatography (2.4.14).
aldehyde-free ethanol (95 per cent) add 0.5 ml of a 0.5 per Test solution. Weigh accurately about 50.0 mg of the substance
cent w/v solution of blue tetrazolium in aldehyde-free ethanol under examination, dissolve in 10 ml of toluene without heating
(95 per cent) and 0.5 ml of a solution prepared by diluting 1 and dilute to 100.0 ml with the mobile phase; dilute 5.0 ml of
volume of tetramethylammonium hydroxide solution (10 per this solution to 50.0 ml with the mobile phase; further dilute
cent) with aldehyde-free ethanol (95 per cent) to make 10 5.0 ml of this solution to 50.0 ml with the mobile phase.
volumes. Allow to stand for exactly 5 minutes and add 1 ml of
Reference solution (a). Dissolve 50.0 mg of ergocalciferol
glacial acetic acid. Measure the absorbance of the resulting
RS in 10 ml of toluene without heating and dilute to 100.0 ml
solution at the maximum at about 525 nm (2.4.7), using as the
with the mobile phase; dilute 5.0 ml of this solution to 50.0 ml
blank a solution prepared by treating 10 ml of aldehyde-free
with the mobile phase; further dilute 5.0 ml of this solution to
ethanol (95 per cent) in the same manner. The absorbance is
not more than that obtained by carrying out the procedure
described above simultaneously using a solution containing Reference solution (b). Dissolve 50.0 mg of cholecalciferol
0.2 µg per ml of hydroquinone in aldehyde-free ethanol (95 RS in 10 ml of toluene without heating and dilute to 100.0 ml
465
ERGOMETRINE MALEATE IP 2007
with the mobile phase. Dilute 5.0 ml of this solution to 50.0 ml Ergometrine Maleate is 9,10-didehydro-N-[(S)-2-hydroxy-1-
with the mobile phase.Heat under a reflux condenser 5.0 ml of methylethyl]-6-methylergoline-8β-carboxamide hydrogen
this solution, under nitrogen, using a water-bath for 60 minutes maleate.
to obtain a solution of cholecalciferl, precholecalciferol and Ergometrine Maleate contains not less than 98.0 per cent and
trans-cholecalciferol. Cool and dilute the refluxed solution to not more than 101.0 per cent of C19H23N3O2,C4H4O4, calculated
50.0 ml with the mobile phase. on the dried basis.
Description. A white or faintly yellow, crystalline powder;
odourless. It is affected by light.
porous silica particles 3 to 10 µm) ( such as Nucleosil
50-S 5 µm), Identification
– mobile phase: a mixture of 997 volumes of hexane and
3 volumes of 1-pentanol, Test A may be omitted if tests B, C, D and E are carried out.
– flow rate. 2 ml per minute, Tests B, D and E may be omitted if tests A and C are carried
– spectrophotometer set at 254 nm, out.
– a 10 or 20 µl loop injector.
Inject a suitable volume of reference solution (b) and adjust Compare the spectrum with that obtained with ergometrine
the sensitivity so that the height of the peak due to maleate RS or with the reference spectrum of ergometrine
cholecalciferol is more than 50 per cent of full-scale maleate.
deflection.The approximate relative retention times calculated
B. Dissolve 30 mg in sufficient 0.01 M hydrochloric acid to
with reference to cholecalciferol are 0.4 for precholecalciferol
produce 100 ml and dilute 10 ml of the solution to 100 ml with
and 0.5 for trans-cholecalciferol. The resolution between
the same solvent. When examined in the range 230 nm to
precholecalciferol and trans-cholecalciferol should not be less
360 nm (2.4.7), the resulting solution shows an absorption
than 1.0; if necessary adjust the proportions of the
maximum at about 311 nm and a minimum at 265 nm to 272 nm;
constituents and flow rate of the mobile phase to obtain the
absorbance at about 311 nm, 0.52 to 0.58.
required resolution.
Inject a suitable volume of reference solution (a) and adjust
the sensitivity so that height of the peak due to ergocalciferol
is more than 50 per cent of full-scale deflection. Inject the
same volume of the test solution. Measure the areas of the D. Dissolve 0.1 g, without heating and protected from light, in
major peaks. sufficient carbon dioxide-free water to produce 10 ml (solution
A). To 0.1 ml of solution A add 1 ml of glacial acetic acid,
Calculate the content of C28H44O.
1 drop of ferric chloride solution and 1 ml of phosphoric acid
Storage. Store protected from light in hermetically-sealed and heat on a water-bath at 80°; a blue or violet colour is
containers under nitrogen in a refrigerator (2° to 8°). The produced after about 10 minutes.
contents of an opened container should be used immediately.
E. To 1 ml of a 0.01 per cent w/v solution add 2 ml of 4-
dimethylaminobenzaldehyde solution; a deep blue colour is
produced after about 10 minutes.
Ergometrine Maleate
Tests
Ergonovine Maleate
Appearance of solution. Solution A is clear (2.4.1), and not
H more intensely coloured than reference solution YS5 or BYS5
O N (2.4.1).
OH
CH3 COOH pH (2.4.24). 3.6 to 4.4, determined in solution A.
, Specific optical rotation (2.4.22). +50.0° to +56.0°, determined
N COOH
CH3 in solution A.
H
Related substances. Carry out the following operations as
rapidly as possible, protected from light.
HN
C19H23N3O2,C4H4O4 Mol.Wt.4415 plate with silica gel G.
466
IP 2007 ERGOMETRINE INJECTION
Mobile phase. A mixture of 75 volumes of chloroform, 25 Description. A clear, colourless or faintly yellow solution.
Identification
Solvent mixture. A mixture of 1 volume of strong ammonia A. In the test for Related substances, the principal spot in the
solution and 9 volumes of ethanol (80 per cent). chromatogram obtained with the test solution corresponds to
that in the chromatogram obtained with the reference solution
Test solution (a). Dissolve 0.1 g of the substance under (a).
B. Exhibits a blue fluorescence.
the same solvent mixture. C. To a volume containing 0.1 mg of Ergometrine Maleate add
0.5 ml of water and 2 ml of 4-dimethylaminobenzaldehyde
Reference solution (a). A 0.1 per cent w/v solution of solution; a deep blue colour is produced after 10 minutes.
ergometrine maleate RS in the same solvent mixture.
Tests
ergometrine maleate RS in the same solvent mixture. pH (2.4.24). 2.7 to 3.5.
Reference solution (c). A 0.005 per cent w/v solution of Related substances. Carry out the following procedure in
ergometrine maleate RS in the same solvent mixture. subdued light and protect from light any solutions not used
immediately.
phase to rise 14 cm. Dry the plate in a current of cold air and Determine by thin-layer chromatography (2.4.17), coating the
spray with 4-dimethylaminobanzaldehyde solution. Dry in a plate with silica gel G slurried with 0.1 M sodium hydroxide.
current of warm air for about 2 minutes. Any secondary spot Mobile phase. A mixture of 90 volumes of chloroform and 10
in the chromatogram obtained with the test solution is not volumes of methanol.
with reference solution (b) and not more than one such spot is Test solution. Evaporate a volume of the injection containing
more intense than the spot in the chromatogram obtained 1 mg of Ergometrine Maleate to dryness at 20° at a pressure
with reference solution (c). not exceeding 2 kPa and dissolve the residue in 0.25 ml of
methanol.
on 0.2 g by drying in an oven at 80° at a pressure not exceeding Reference solution (a). A 0.4 per cent w/v solution of
2.7 kPa for 2 hours. ergometrine maleate RS in methanol.
Assay. Weigh accurately about 0.15 g and dissolve in 40 ml of Reference solution (b). Dilute 5 ml of reference solution (a) to
anhydrous glacial acetic acid. Titrate with 0.05 M perchloric 50 ml with methanol.
acid, determining the end-point potentiometrically (2.4.25). Reference solution (c). Dilute 5 ml of reference solution (b) to
Carry out a blank titration. 10 ml with methanol.
1 ml of 0.05 M perchloric acid is equivalent to 0.02207 g of Reference solution (d). Dilute 5 ml of reference solution (c) to
C19H23N3O2,C4H4O4. 10 ml with methanol.
Storage. Store protected from light in a refrigerator (2° to 8°). Apply to the plate 5 µl of each solution. After development,
Assess the intensities of any secondary spots in the
chromatogram obtained with the test solution by reference to
the spots in the chromatograms obtained with reference
Ergometrine Injection solutions (b), (c) and (d). The total of the intensities so
Ergometrine Maleate Injection; Ergonovine Injection, assessed does not exceed 10 per cent of the intensity of the
Ergonovine Maleate Injection principal spot.
Ergometrine Injection is a sterile solution of Ergometrine Other tests. Complies with the tests stated under Parenteral
Maleate in Water for Injections containing suitable stabilising Preparations (Injections).
agents. Assay. Carry out the following procedure protected from
Ergometrine Injection contains not less than 90.0 per cent and light.
not more than 110.0 per cent of the stated amount of Dilute a suitable volume, accurately measured, of the injection
ergometrine maleate, C19H23N3O2,C4H4O4. with sufficient water to produce a solution containing
467
ERGOMETRINE TABLETS IP 2007
0.004 per cent w/v of Ergometrine Maleate. To 3.0 ml add 6.0 ml centrifuge and remove the supernatant liquid. Extract the
of 4-dimethylaminobenzaldehyde solution, mix, cool to room residue with two quantities, each of 1 ml, of methanol,
temperature and allow to stand for 30 minutes (solution A). At evaporate the combined extracts to dryness at 20° at a pressure
the same time prepare solution B in the same manner but using not exceeding 2 kPa and dissolve the residue in 0.25 ml of
3.0 ml of a 0.004 per cent w/v solution of ergometrine maleate methanol, centrifuge if necessary.
RS and beginning at the words “add 6.0 ml......”. Measure the Reference solution (a). A 0.4 per cent w/v solution of
absorbance of solution B at the maximum at about 545 nm ergometrine maleate RS in methanol.
(2.4.7), using as the blank a solution prepared by mixing 6.0 ml
of 4-dimethylaminobenzaldehyde solution and 3.0 ml of water. Reference solution (b). Dilute 5 ml of reference solution (a) to
Without delay replace solution B with solution A, using the 50 ml with methanol.
same cell, and measure the absorbance of solution A at the Reference solution (c). Dilute 5 ml of reference solution (b) to
same wavelength. Calculate the content of C19H23N3O2,C4H4O4. 10 ml with methanol.
Storage. Store protected from light in single dose containers Reference solution (d). Dilute 5 ml of reference solution (c) to
in a refrigerator (2° to 8°). 10 ml with methanol.
Ergometrine Tablets Assess the intensities of any secondary spots in the
chromatogram obtained with the test solution by reference to
Ergometrine Maleate Tablets; Ergonovine Tablets; the spots in the chromatograms obtained with reference
Ergonovine Maleate Tablets solutions (b), (c) and (d). The total of the intensities so
assessed does not exceed 10 per cent of the intensity of the
Ergometrine Tablets contain not less than 90.0 per cent and
principal spot.
ergometrine maleate, C19H23N3O2,C4H4O4. The tablets may be Uniformity of content. Comply with the test stated under
coated. Tablets.
Protect the solutions from light throughout the test.
Identification
To one tablet add 10.0 ml of a 1 per cent w/v solution of
A. In the test for Related substances, the principal spot in the tartaric acid, shake for 30 minutes, centrifuge and use the
chromatogram obtained with the test solution corresponds to supernatant liquid. Dilute a suitable volume, accurately
that in the chromatogram obtained with the reference solution measured, with sufficient water to produce a solution
(a). containing 0.004 per cent w/v of Ergometrine Maleate. To 3.0
B. Extract a quantity of the powdered tablets containing 2 mg ml add 6.0 ml of 4-dimethylaminobenzaldehyde solution, mix,
of Ergometrine Maleate with 20 ml of water, filter and wash the cool to room temperature and allow to stand for 30 minutes
residue with sufficient water to produce 20 ml. The solution (solution A). At the same time prepare solution B in the same
exhibits a blue fluorescence. manner but using 3.0 ml of a 0.004 per cent w/v solution of
ergometrine maleate RS and beginning at the words “add 6.0
C. To 2 ml of the solution obtained in test B add 4 ml of 4- ml....” Measure the absorbance of solution B at the maximum
dimethylaminobenzaldehyde solution; a deep blue colour is at about 545 nm (2.4.7), using as the blank a solution prepared
produced after about 10 minutes. by mixing 6.0 ml of 4-dimethylaminobenzaldehyde solution
and 3.0 ml of water. Without delay replace solution B with
Tests
solution A, using the same cell, and measure the absorbance
Related substances. Carry out the following procedure in of solution A at the same wavelength. Calculate the content of
subdued light and protect from light any solutions not used C19H23N3O2,C4H4O4 in the tablet.
immediately. Other tests. Comply with the tests stated under Tablets.
Determine by thin-layer chromatography (2.4.17), coating the Assay. Weigh and powder 20 tablets. Weigh accurately a
plate with silica gel G slurried with 0.1 M sodium hydroxide. quantity of the powder containing about 2 mg of Ergometrine
Mobile phase. A mixture of 90 volumes of chloroform and 10 Maleate, shake with 50.0 ml of a 1 per cent w/v solution of
volumes of methanol. tartaric acid for 30 minutes, centrifuge and use the supernatant
liquid. Carry out the procedure described under Uniformity of
content beginning at the words “To 3.0 ml add 6 ml.....”.
containing 1 mg of Ergometrine Maleate with 0.2 ml of a 1 per
cent w/v solution of domiphen bromide, add 2 ml of methanol, Storage. Store protected from light.
468
IP 2007 ERGOTAMINE TARTRATE
Ergotamine Tartrate the red tinge becomes less apparent and the blue colour more
pronounced.
H E. Dissolve 1 mg in 5 ml of a 1 per cent w/v solution of tartaric

O N O OH acid. To 1 ml of this solution add slowly 3 ml of
4-dimethylaminobenzaldehyde solution and mix; a deep blue
N
H3C N H OH
colour is produced.
, COOH
O
N O HOOC
CH3 OH Tests
H
Carry out the following tests as rapidly as possible, protected
from light.
HN
Appearance of solution. Mix 50 mg with 25 mg of tartaric acid
(C33H35N5O5)2,C4H6O6 Mol. Wt. 1313.4 and dissolve in 20 ml of water. The solution is clear (2.4.1),
and not more intensely coloured than reference solution YS6
Ergotamine Tartrate is (5'S)-12'-hydroxy-2'-methyl-3',6',18-
(2.4.1).
trioxo-5-benzylergotaman (2R,3R)tartrate.
Ergotamine Tartrate contains not less than 98.0 per cent and
suspension.
not more than 101.0 per cent of (C33H35N5O5)2,C4H 6O6,
calculated on the dried basis. Specific optical rotation (2.4.22). The specific optical rotation
Description. Colourless crystals, or a white or almost white, of the ergotamine base, checked for purity by the method
crystalline powder; odourless; slightly hygroscopic. It may given below, is –154 ° to –165°, determined by the following
contain two molecular equivalents of methanol of method. Dissolve 0.4 g in 40 ml of a 1 per cent w/v solution of
crystallisation. tartaric acid, cautiously add 0.5 g of sodium bicarbonate in
small portions and mix well. Wash 100 ml of chloroform by
Identification shaking with 5 quantities, each of 50 ml, of water and extract
the solution of the substance under examination with
Test A may be omitted if tests B, C, D and E are carried out. 4 quantities, each of 10 ml, of the washed chloroform. Filter
Tests B, C and D may be omitted if tests A and E are carried the combined chloroform extracts through a small filter
out. moistened with the washed chloroform, dilute to 50 ml with
A. Determine by infrared absorption spectrophotometry (2.4.6). the same solvent and measure the optical rotation.
Before triturating with potassium bromide IR during To 25 ml of the chloroform solution add 50 ml of anhydrous
preparation of the disc, triturate first with 0.2 ml of methanol. glacial acetic acid. Titrate with 0.05 M perchloric acid,
Compare the spectrum with that obtained with ergotamine determining the end-point potentiometrically (2.4.25). Carry
tartrate RS or with the reference spectrum of ergotamine out a blank titration.
tartrate.
B. When examined in the range 230 nm to 360 nm (2.4.7), a ergotamine base, C33H35N5O5.
0.005 per cent solution in 0.01 M hydrochloric acid shows an
absorption maximum at 311 nm to 321 nm and a minimum at Related substances. Determine by thin-layer chromatography
265 nm to 275 nm; absorbance at the maximum, 0.59 to 0.64, (2.4.17), coating the plate with silica gel G.
calculated on the dried basis. Mobile phase. A mixture of 70 volumes of ether, 15 volumes of
C. In the test for Related substances, the principal spot in the dimethylformamide, 10 volumes of chloroform and 5 volumes
chromatogram obtained with test solution (b) corresponds to of ethanol.
that in the chromatogram obtained with reference solution (a) Prepare the following solutions immediately before use in
when examined for not more than 1 minute in ultraviolet light the order stated.
at 365 nm or when examined in daylight after spraying with
ethanolic 4-dimethylaminobenzaldehyde solution. Solvent mixture. A mixture of 9 volumes of chloroform and 1
volume of methanol.
D. Dissolve 1 mg in a mixture of 5 ml of glacial acetic acid and
5 ml of ethyl acetate. To 1 ml of the solution add 1 ml of Test solution (a). Dissolve 0.1 g of the substance under
sulphuric acid, with continuous shaking and cooling; a blue examination in 10 ml with solvent mixture.
colour with a red tinge develops. Add 0.1 ml of ferric chloride Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with
test solution previously diluted with an equal volume of water; the same solvent mixture.
469
ERGOTAMINE INJECTION IP 2007
Reference solution (a). A 0.1 per cent w/v solution of Identification

ergotamine tartrate RS in the same solvent mixture.
A. In the test for Ergot alkaloids and related substances, the
Reference solution (b). A 0.015 per cent w/v solution of principal spot in the chromatogram obtained with the test
ergotamine tartrate RS in the same solvent mixture. solution corresponds to that due to ergotamine in the
Reference solution (c). A 0.005 per cent w/v solution of chromatogram obtained with the reference solution.
ergotamine tartrate RS in the same solvent mixture. B. To a volume containing 0.2 mg of Ergotamine Tartrate add 1
Apply to the plate 5 µl of each solution. Immediately after ml of 4-dimethylaminobenzaldehyde solution; a deep blue
application expose the plate to an atmosphere saturated with colour is produced.
ammonia vapour for exactly 20 seconds, dry the plate at the C. Mix a volume containing 2 mg of Ergotamine Tartrate with 2
line of application in a current of cold air and immediately start ml of dilute sulphuric acid, dissolve a few mg of magnesium
developing the chromatogram, allowing the mobile phase to powder in the solution and add 25 mg of resorcinol. Shake to
rise 17 cm. Dry the plate in a current of dry air for 2 minutes dissolve, carefully add 2 ml of sulphuric acid down the inside
and examine in ultraviolet light at 365 nm for not more than of the tube and warm gently; a red ring forms at the interface
1 minute. Spray abundantly with ethanolic 4- of the two liquid layers and spreads throughout the lower
dimethylaminobenzaldehyde solution and dry in a current of layer.
warm air for about 2 minutes. Any secondary spot in the
chromatogram obtained with test solution (a) is not more Tests
intense than the spot in the chromatogram obtained with Carry out the following tests as rapidly as possible, protected
reference solution (b) and not more than one such spot is from light.
pH (2.4.24). 2.8 to 3.8.
with reference solution (c).
Ergot alkaloids and related substances. Determine by thin-
Loss on drying (2.4.19). Not more than 6.0 per cent, determined layer chromatography (2.4.17), coating the plate with silica
on 0.1 g by drying over phosphorus pentoxide at a pressure gel G slurried with 0.1 M sodium hydroxide.
of 1.5 to 2.5 kPa for 6 hours.
Mobile phase. A mixture of 90 volumes of chloroform and 10
Assay. Weigh accurately about 0.2 g and dissolve in 40 ml of volumes of methanol.
anhydrous glacial acetic acid Titrate with 0.05 M perchloric
Test solution. Add sufficient of a 10 per cent w/v solution of
sodium bicarbonate to a volume of the injection containing 5
mg of Ergotamine Tartrate to make it distinctly alkaline to litmus
1 ml of 0.05 M perchloric acid is equivalent to 0.03284 g of paper. Extract with five quantities, each of 10 ml, of chloroform,
(C33H35N5O5)2,C4H6O6. filter the extracts through a small double filter paper, wash the
Storage. Store protected from light in sealed glass containers, filter with chloroform, evaporate the combined filtrates and
under nitrogen and in a refrigerator (2° to 8°). washings to dryness at 20° at a pressure of about 1.5 kPa and
dissolve the residue in 1 ml of a mixture of equal volumes of
chloroform and methanol.
Reference solution. Dissolve 5 mg of ergotamine tartrate RS
Ergotamine Injection in 10 ml of a 1 per cent w/v solution of tartaric acid and
complete the preparation described for the test solution
Ergotamine Tartrate Injection beginning at the words “Extract with five quantities...”.
Ergotamine Injection is a sterile solution of Ergotamine Tartrate Apply without delay, to the plate 20 µl of the test solution and
in Water for Injection containing Ethanol (95 per cent), Glycerin 14 µl, 10 µl, 7 µl and 2 µl of the reference solution. After
and sufficient Tartaric Acid to adjust the pH of the solution to development, dry the plate in air and examine in ultraviolet
3.3. light at 365 nm. The chromatogram obtained with the test
solution shows two principal spots, corresponding to
Ergotamine Injection contains a quantity of total alkaloids,
ergotamine and, of higher Rf value, ergotaminine; a spot
calculated as (C33H35N5O5)2,C4H6O6, equivalent to not less than
between the two principal spots and a number of spots of
lower Rf values may also be seen. Compare the chromatogram
amount of ergotamine tartrate, of which 50 to 70 per cent is
obtained with the test solution with the chromatograms
present as ergotamine tartrate.
obtained with the reference solution. The spot corresponding
Description. A clear, colourless or almost colourless solution. to ergotaminine is not larger or more intense than the spot
470
IP 2007 ERGOTAMINE TABLETS
corresponding to ergotamine obtained with 7 µl of the the residue add 10 ml of chloroform saturated with strong
reference solution. The spot corresponding to ergotamine is ammonia solution, triturate, filter and evaporate the filtrate to
not smaller or less intense than the spot corresponding to dryness on a water-bath. The residue so obtained complies
ergotamine obtained with 10 µl of the reference solution and with the following tests.
is not larger or more intense than the spot corresponding to
Dissolve 1 mg in a mixture of 5 ml of glacial acetic acid and 5
ergotamine obtained with 14 µl of the reference solution,
ml of ethyl acetate. To 1 ml of the solution add 1 ml of sulphuric
corresponding to not less than 50 per cent and not more than
acid, with continuous shaking and cooling; a blue colour
70 per cent of ergotamine tartrate. Any other spots are not
with a red tinge develops. Add 0.1 ml of ferric chloride test
larger or more intense than the spot corresponding to
solution previously diluted with an equal volume of water;
ergotamine obtained with 2 µl of the reference solution.
the red tinge becomes less apparent and the blue colour more
Other tests. Complies with the tests stated under Parenteral pronounced.
B. Dissolve 1 mg in 5 ml of a 1 per cent w/v solution of tartaric
Assay. To an accurately measured volume add sufficient of a acid. To 1 ml of this solution add slowly 3 ml of
0.25 per cent w/v solution of tartaric acid to produce a solution 4-dimethylaminobenzaldehyde solution and mix; a deep blue
containing about 0.005 per cent w/v of Ergotamine Tartrate. colour is produced.
Mix 3.0 ml of this solution with 6.0 ml of
4-dimethylaminobenzaldehyde solution, cool to room
temperature and allow to stand for 30 minutes (solution A). At
the same time mix 3.0 ml of a 0.003 per cent w/v solution of
ergometrine maleate RS in a 0.25 per cent w/v solution of Tests
tartaric acid with 6.0 ml of 4-dimethylaminobenzaldehyde
solution, cool to room temperature and allow to stand for 30 Carry out the following tests as rapidly as possible, protected
minutes (solution B). Prepare solution C by mixing 3.0 ml of a from light.
0.25 per cent w/v solution of tartaric acid with 6.0 ml of 4-
dimethylaminobenzaldehyde solution. Measure the
absorbance of solution B at 545 nm (2.4.7), using solution C as
the blank. Without delay replace solution B with solution A, Mobile phase. A mixture of 70 volumes of ether, 15 volumes of
using the same cell, and measure the absorbance of solution dimethylformamide, 10 volumes of chloroform and 5 volumes
A at the same wavelength. Calculate the content of total of ethanol.
alkaloids as (C33H35N5O5)2,C4H6O6 from the absorbances Prepare the following solutions immediately before use in
obtained. the order stated.
1 mg of ergometrine maleate RS is equivalent to 1.488 mg of Test solution. Extract a quantity of the powdered tablets
(C33H35N5O5)2,C4H6O6. containing 1 mg of Ergotamine Tartrate with 2 ml of a mixture
Storage. Store protected from light in single dose glass of equal volumes of chloroform and methanol and centrifuge.
containers at a temperature not exceeding 30°. Remove the supernatant liquid, extract the residue with two
quantities, each of 1 ml, of the solvent mixture, evaporate the
combined extracts to dryness at 20° at a pressure of 2 kPa and
dissolve the residue in 0.25 ml of a mixture of equal volumes of
chloroform and methanol; centrifuge if necessary.
Ergotamine Tablets
Ergotamine Tartrate Tablets ergotamine tartrate RS in the same solvent mixture.
Ergotamine Tablets contain not less than 90.0 per cent and Reference solution (b). A 0.04 per cent w/v solution of
not more than 110.0 per cent of the stated amount of ergotamine tartrate RS in the same solvent mixture.
ergotamine tartrate, (C33H35N5O5)2,C4H6O6. The tablets may be
coated. Reference solution (c). A 0.02 per cent w/v solution of
Identification Reference solution (d). A 0.01 per cent w/v solution of
A. Triturate a quantity of the powdered tablets containing 5
mg of Ergotamine Tartrate with 10 ml of light petroleum (40° Apply to the plate 5 µl of each solution. Immediately after
to 60°), allow to settle and discard the petroleum extract. To application expose the plate to an atmosphere saturated with
471
ERYTHROMYCIN IP 2007
ammonia vapour for exactly 20 seconds, dry the plate at the Erythromycin
line of application in a current of cold air and immediately start
developing the chromatogram, allowing the mobile phase to O
rise 17 cm. Dry the plate in air and examine in ultraviolet light H3C CH3
at 365 nm. Assess the intensity of any secondary spots in the R 1 OH
chromatogram obtained with the test solution by reference to H3C OH CH3
H3C
the spots in the chromatograms obtained with reference O Erythromycin R1 R2
H3C
solutions (a), (b) and (c). The sum of the intensities so assessed O O CH3 O A OH CH3
B H CH3
in the chromatogram obtained with the test solution should CH3 O
C OH H
O N(CH3)2
not exceed 10 per cent of the intensity of the principal spot in OH
the chromatogram obtained with the test solution. In addition, OR2
OH
any single secondary spot in the chromatogram obtained with CH3
chromatogram obtained with reference solution (d). C37H67NO13 Mol. Wt. 733.9
Uniformity of content. Comply with the test stated under Erythromycin is a mixture of macrolide antibiotics consisting
Tablets. largely of erythromycin A,
To one tablet add sufficient quantity of a 1 per cent w/v solution (2R,3S,4S,5R,6R,8R,10R,11R,12S,13R)-5-(3-amino-3,4,6-
of tartaric acid to produce a solution containing 0.05 mg of trideoxy-N,N-dimethyl-α-D-xylo-hexopyranosyloxy)-3-(2,6-
Ergotamine Tartrate per ml. Shake for 30 minutes and centrifuge. dideoxy-3-C,3-O- dimethyl-α-L-ribo-hexopyranosyloxy)-13-
Mix 3.0 ml of this solution with 6.0 ml of ethyl-6,11,12- trihydroxy-2,4,6,8,10,12-hexamethyl-9-
4-dimethylaminobenzaldehyde solution, cool to room oxotridecan-13-olide, it is produced by the growth of certain
temperature and allow to stand for 30 minutes (solution A). At strains of Streptomyces erythreus.
the same time mix 3.0 ml of a 0.003 per cent w/v solution of Erythromycin has a potency not less than 920 Units per mg,
ergometrine maleate RS in a 0.25 per cent w/v solution of calculated on the anhydrous basis.
tartaric acid with 6.0 ml of 4-dimethylaminobenzaldehyde
solution, cool to room temperature and allow to stand for 30 Description. Colourless or slightly yellow crystals or a white
minutes (solution B). Prepare solution C by mixing 3.0 ml of a or slightly yellow powder; slightly hygroscopic.
0.25 per cent w/v solution of tartaric acid with 6.0 ml of 4-
dimethylaminobenzaldehyde solution. Measure the
Identification
absorbance of solution B at 545 nm (2.4.7), using solution C as Test A may be omitted if tests B, C and D are carried out. Tests
the blank. Without delay replace solution B with solution A, B and D may be omitted if tests A and C are carried out.
using the same cell, and measure the absorbance of solution
A at the same wavelength. Calculate the content of total A. Determine by infrared absorption spectrophotometry (2.4.6).
alkaloids as (C33H35N5O5)2,C4H6O6 from the absorbances Compare the spectrum with that obtained with erythromycin
obtained. RS or with the reference spectrum of erythromycin.
1 mg of ergometrine maleate RS is equivalent to 1.488 mg of B. Determine by thin-layer chromatography (2.4.17), coating

(C33H35N5O5)2,C4H6O6. the plate with silica gel G.
Calculate the content of (C33H35N5O5)2,C4H6O6 in the tablet. Mobile phase. The upper layer obtained by shaking together
45 volumes of ethyl acetate, 40 volumes of a 15 per cent w/v
Other tests. Comply with the tests stated under Tablets. solution of ammonium acetate previously adjusted to pH 9.6
Assay. Weigh and powder 20 tablets. Weigh accurately a with 10 M ammonia and 20 volumes of 2-propanol and allowing
quantity of the powder containing about 5 mg of Ergotamine to separate.
Tartrate and dissolve in 50 ml of a 1 per cent w/v solution of Test solution. Dissolve 0.1 g of the substance under
tartaric acid, allow to stand for 30 minutes with frequent examination in 100 ml of methanol.
shaking and dilute to 100.0 ml with water. Using 3.0 ml of the
clear supernatant liquid, carry out the procedure described Reference solution (a). A 0.1 per cent w/v solution of
under Uniformity of content beginning at the words “Mix 3.0 erythromycin RS in methanol.
ml of this solution with 6.0 ml of 4-dimethylaminobenzaldehyde Reference solution (b). A 0.2 per cent w/v solution of
solution....”. spiramycin RS in methanol.
Storage. Store protected from light at a temperature not Apply to the plate 10 µl of each solution. After development,
exceeding 30°. dry the plate in air, spray with ethanolic anisaldehyde
472
IP 2007 ERYTHROMYCIN TABLETS
solution, heat at 110° for 5 minutes and allow to cool. The Storage. Store protected from light at a temperature not
principal spot in the chromatogram obtained with the test exceeding 30°.
with reference solution (a) and is different in position and
colour from the spots in the chromatogram obtained with
reference solution (b). Erythromycin Tablets
C. To about 5 mg add 5 ml of a 0.02 per cent w/v solution of Erythromycin Tablets contain not less than 90.0 per cent and
xanthydrol in a mixture of 1 volume of hydrochloric acid and not more than 110.0 per cent of the stated amount of
99 volumes of 5 M acetic acid and heat on a water-bath; a red erythromycin, C37H67NO13. The tablets are enteric-coated.
colour is produced.
Identification
D. Dissolve about 10 mg in 5 ml of 7 M hydrochloric acid and
allow to stand for about 20 minutes; a yellow colour develops. A. Shake a quantity of the powdered tablets containing 0.1 g
of Erythromycin with 5 ml of chloroform, decolorise if
Tests necessary, with decolorising charcoal, filter and evaporate
the filtrate to dryness. The residue after drying at a pressure
pH (2.4.24). 8.0 to 10.5, determined in a 0.066 per cent w/v not exceeding 0.7 kPa complies with the following test.
solution in carbon dioxide-free water.
Specific optical rotation (2.4.22). –71.0° to –78.0°, determined Compare the spectrum with that obtained with erythromycin
in a 2.0 per cent w/v solution in ethanol. Measure the optical RS or with the reference spectrum of erythromycin.
rotation at least 30 minutes after preparing the solution.
B. Dissolve a quantity of the powdered tablets containing 3
Related substances. Determine by thin-layer chromatography mg of Erythromycin as completely as possible in 2 ml of
(2.4.17), coating the plate with silanised silica gel H. acetone and add 2 ml of hydrochloric acid; an orange colour
Mobile phase. A mixture of 75 volumes of methanol and 45 is produced which changes to red and then to deep purplish
volumes of a 5 per cent w/v solution of ammonium acetate. red. Add 2 ml of chloroform and shake; the chloroform layer
becomes purple.
examination in 100 ml of methanol. Tests
erythromycin RS in methanol.
manner.
Weigh and powder 20 tablets. Weigh accurately a quantity of
dry the plate in air, spray with ethanolic anisaldehyde
the powder containing about 0.4 g of Erythromycin and
solution, heat at 110° for 5 minutes and allow to cool. Any
triturate with 10 ml of sterile phosphate buffer pH 8.0 and add
secondary spot with an Rf value lower than that of the principal
sufficient sterile phosphate buffer pH 8.0 to produce 100.0 ml.
spot in the chromatogram obtained with the test solution is
Calculate the content of erythromycin in the tablets, taking
not more intense than the principal spot in the chromatogram
each 1000 Units found to be equivalent to 1 mg of
erythromycin.
exceeding 30°.
Water (2.3.43). Not more than 6.5 per cent, determined on 0.2
g using a 10 per cent w/v solution of imidazole in anhydrous
methanol as the solvent.
Method A (2.2.10), using a solution prepared by dissolving
about 25 mg, accurately weighed, in 10 ml of methanol and
adding sufficient water to produce 100.0 ml. Express the results
as units per mg.
473
ERYTHROMYCIN ESTOLATE IP 2007
Erythromycin Estolate Tests

pH (2.4.24). 5.5 to 7.0, determined in the supernatant liquid
obtained by suspending 0.4 g in 10 ml of carbon dioxide-free
O
H3C water, shaking for 5 minutes and allowing to stand.
CH3
1 OH Related substances. Determine by thin-layer chromatography
R
OH CH3 (2.4.17), coating the plate with silanised silica gel G.
H3C
H3C
O Mobile phase. A mixture of 85 volumes of chloroform, 5
H3C volumes of ethanol (95 per cent) and 1 volume of a 15 per
O O CH3 O , C12H25OSO3H cent w/v solution of ammonium acetate previously adjusted
CH3 O to pH 7.0.
O N(CH3)2
OH Test solution (a). Dissolve 0.4 g of the substance under
2
OR examination in 100 ml of acetone.
OH
CH3
Erythromycin R1 R2 acetone.
A OH CH3 Reference solution (a). A 0.1 per cent w/v solution of
B H CH3 erythromycin estolate RS in acetone.
C OH H
w/v each of erythromycin estolate RS and erythromycin
C40H71NO14,C12H26O4S Mol. Wt. 1056.4 ethylsuccinate RS in acetone.
Erythromycin Estolate is Reference solution (c). A 0.008 per cent w/v of erythromycin
(2R,3S,4S,5R,6R,8R,10R,11R,12S,13R)-5-(3-amino-3,4,6- RS in acetone.
trideoxy-N,N-dimethyl-β-D-xylo-hexopyranosyloxy)-3-(2,6-
dideoxy-3-C,3-O-dimethyl-α-L-ribo-hexopyranosyloxy)-13- Apply to the plate 10 µl of each solution. After development,
ethyl-6,11,12-trihydroxy-2,4,6,8,10,12-hexamethyl-9- dry the plate in air, spray with anisaldehyde solution, heat at
oxotridecan-13-olide dodecyl sulphate. 110° for 5 minutes and allow to cool. Any secondary spot in
Erythromycin Estolate has a potency not less than 610 Units intense than the principal spot in the chromatogram obtained
per mg, calculated on the anhydrous basis. with reference solution (c).
Description. A white, crystalline powder. Content of dodecyl sulphate, C12H26O4S. 23.0 per cent to 25.5
per cent, calculated on the anhydrous basis and determined
Identification by the following method. Weigh accurately about 0.5 g and
Test A may be omitted if tests B, C and D are carried out. Tests dissolve in 25 ml of dimethylformamide. Titrate with 0.1 M
B and C may be omitted if tests A and D are carried out. sodium methoxide, using 0.05 ml of a 0.3 per cent w/v solution
of thymol blue in methanol as indicator. Carry out a blank
titration.
Compare the spectrum with that obtained with erythromycin
estolate RS or with the reference spectrum of erythromycin 1 ml of 0.1 M sodium methoxide is equivalent to 0.02664 g of
estolate. C12H26O4S.
B. In the test for Related substances, the principal spot in the Sulphated ash (2.3.18). Not more than 0.5 per cent, determined
chromatogram obtained with test solution (b) corresponds to on 0.5 g.
that in the chromatogram obtained with reference solution (a). Water (2.3.43). Not more than 4.0 per cent, determined on 0.3
The test is not valid unless the chromatogram obtained with g using a 10 per cent w/v solution of imidazole in anhydrous
reference solution (b) shows two clearly separated principal methanol as the solvent.
spots.
C. Suspend about 3 mg in 2 ml of 1 M sulphuric acid, add 0.1
ml of a 0.01 per cent w/v solution of methylene blue and 2 ml
manner. Weigh accurately about 40 mg, dissolve in 40 ml of
of chloroform and shake; the chloroform layer becomes blue.
methanol and add 20 ml of phosphate buffer pH 7.0 and
D. Dissolve about 10 mg in 5 ml of 7 M hydrochloric acid and sufficient water to produce 100.0 ml. Maintain the solution at
allow to stand for 20 minutes; a yellow colour is produced. 60° for 3 hours and cool. Express the results as units per mg.
474
IP 2007 ERYTHROMYCIN STEARATE
Storage. Store protected from light at a temperature not Erythromycin Stearate has a potency not less than 600 Units
exceeding 30°. per mg, calculated on the anhydrous basis.
Description. Colourless or slightly yellow crystals or a white
or slightly yellow, crystalline powder.
Erythromycin Estolate Tablets
Identification
Erythromycin Estolate Tablets contain not less than 90.0 per
cent and not more than 115.0 per cent of the stated amount of A. Determine by thin-layer chromatography (2.4.17), coating
erythromycin, C37H67NO13. The tablets may be coated. the plate with silica gel G.
Identification Mobile phase. The upper layer of a mixture of 45 volumes of

ethyl acetate, 40 volumes of a 15 per cent w/v solution of
A. To a quantity of the powdered tablets containing 0.1 g of ammonium acetate, previously adjusted to pH 9.6 with 9 M
erythromycin add 10 ml of chloroform, shake well, decolorise ammonia, and 20 volumes of 2-propanol.
if necessary, with decolorising charcoal, filter and evaporate
the filtrate to dryness. The residue after drying at a pressure
not exceeding 0.7 kPa complies with the following test.
Compare the spectrum with that obtained with erythromycin
estolate RS or with the reference spectrum of erythromycin Reference solution (b). A 0.1 per cent w/v solution of stearic
estolate. acid in methanol.
B. Dissolve a quantity of the powdered tablets containing Apply to the plate 5 µl of each solution. After development,
12 mg of erythromycin as completely as possible in 2 ml of dry the plate in air, spray with a solution containing 0.02 per
acetone and add 2 ml of hydrochloric acid; an orange-red cent w/v of 2,7-dichlorofluorescein and 0.01 per cent w/v of
colour is produced which changes to red and then to deep rhodamine B in ethanol (95 per cent), allow the plate to stand
purple. Add 2 ml of chloroform and shake; the chloroform for a few seconds in the vapour above a water-bath and examine
layer becomes purple. in ultraviolet light at 365 nm. The chromatogram obtained with
the test solution exhibits two spots, one of which corresponds
Tests in position to the principal spot in the chromatogram obtained
Other tests. Comply with the tests stated under Tablets. with reference solution (a) and the other to the principal spot
in the chromatogram obtained with reference solution (b).
Assay. Determine by the microbiological assay of antibiotics, Spray the plate with ethanolic anisaldehyde solution, heat at
Method A (2.2.10) on a solution prepared in the following 110° for 5 minutes and examine in daylight. The coloured spot
manner. in the chromatogram obtained with the test solution
Weigh and powder 20 tablets. Weigh accurately a quantity of corresponds to the principal spot in the chromatogram
the powder containing 25 mg of erythromycin, dissolve in obtained with reference solution (a).
40 ml of methanol, add 20 ml of phosphate buffer pH 7.0 and B. To about 5 mg add 5 ml of a 0.02 per cent w/v solution of
sufficient water to produce 100.0 ml. Maintain the solution at xanthydrol in a mixture of 1 volume of hydrochloric acid and
60° for 3 hours, cool and filter. Calculate the content of 99 volumes of 5 M acetic acid and heat on a water-bath; a red
erythromycin in the tablets, taking each 1000 Units found to colour is produced.
be equivalent to 1 mg of erythromycin.
C. Dissolve about 10 mg in 5 ml of 7 M hydrochloric acid and
Storage. Store at a temperature not exceeding 30°. allow to stand for about 20 minutes; a yellow colour develops.
equivalent amount of erythromycin. Tests
suspension.
Erythromycin Stearate Related substances. Determine by thin-layer chromatography
C37H67NO13,C18H36O2 Mol. Wt. 1018.4 (2.4.17), coating the plate with silanised silica gel H.
Erythromycin Stearate is a mixture of the stearate of Mobile phase. A mixture of 100 volumes of methanol and 60
Erythromycin with an excess of stearic acid. volumes of a 15 per cent w/v solution of ammonium acetate.
475
ERYTHROMYCIN STEARATE TABLETS IP 2007
Test solution. Dissolve 0.28 g of the substance under about 50 mg accurately weighed in sufficient methanol to
examination in 100 ml of methanol. produce 100.0 ml. Express the results as units per mg.
Reference solution (a). A 0.2 per cent w/v solution Storage. Store protected from light at a temperature not
erythromycin RS in methanol. exceeding 30°.
Apply to the plate 10 µl of each solution. After development, Erythromycin Stearate Tablets
dry the plate in air, spray with ethanolic anisaldehyde
Erythromycin Stearate Tablets contain not less than 90.0 per
solution, heat at 110° for 5 minutes and allow to cool. Any
spot with an Rf value lower than that of the principal spot in
erythromycin, C37H67NO13. The tablets may be coated.
intense than the corresponding spot in the chromatogram Identification
obtained with reference solution (a) and any spot with an
Rf value higher than that of the principal spot is not more A. To a quantity of the powdered tablets containing 0.1 g of
intense than the principal spot in the chromatogram obtained erythromycin add 10 ml of water and shake well. Decant the
with reference solution (b). supernatant liquid and discard. Extract the residue by shaking
Erythromycin stearate. Not less than 84.0 per cent of with 10 ml of methanol, filter the extract and evaporate to
C37H67NO13,C18H36O2, calculated on the anhydrous basis and dryness. The residue after drying at a pressure not exceeding
determined by the following method. Weigh accurately about 0.7 kPa complies with the following test.
0.5 g and dissolve in 30 ml of chloroform. If the solution is Determine by infrared absorption spectrophotometry (2.4.6).
opalescent, filter and shake the residue with three quantities, Compare the spectrum with that obtained with erythromycin
each of 25 ml, of chloroform. Filter, if necessary, and wash the stearate RS or with the reference spectrum of erythromycin
filter with chloroform. Evaporate the combined filtrate and stearate.
washings on a water-bath to about 30 ml, add 50 ml of
B. Dissolve a quantity of the powdered tablets containing
3 mg of erythromycin as completely as possible in 2 ml of
acetone and add 2 ml of hydrochloric acid; an orange colour
is produced which changes to red and then to deep purplish
1 ml of 0.1 M perchloric acid is equivalent to 0.1018 g of red. Add 2 ml of chloroform and shake; the chloroform layer
C37H67NO13,C18H36O2. becomes purple.
Free stearic acid. Not more than 14.0 per cent of C18H36O2, C. Extract a quantity of the powdered tablets containing 50 mg
calculated on the anhydrous basis and determined by the of erythromycin with 10 ml of chloroform, filter and evaporate
following method. Weigh accurately about 0.4 g and dissolve to dryness. Heat 0.1 g of the residue gently with 5 ml of 2 M
in 50 ml of methanol. Titrate with 0.1 M sodium hydroxide, hydrochloric acid and 10 ml of water until the solution boils;
determining the end-point potentiometrically (2.4.25). Calculate oily globules rise to the surface. Cool, remove the fatty layer,
the volume of 0.1 M sodium hydroxide required for each g of heat it with 3 ml of 0.1 M sodium hydroxide and allow to cool;
the substance and subtract the volume of 0.1 M perchloric the solution sets to a gel. Add 10 ml of hot water and shake;
acid required for each g of the substance in the test for the solution froths. To 1 ml add a 10 per cent w/v solution of
Erythromycin stearate. calcium chloride; a granular precipitate is produced which is
1 ml of the difference is equivalent to 0.02845 g of C18H36O2. insoluble in hydrochloric acid.
Erythromycin stearate and free stearic acid. 98.0 to 103.0 per Tests
cent, calculated by adding together the percentages of
erythromycin stearate and free stearic acid determined as Disintegration (2.5.1). 90 minutes.
described above. Other tests. Comply with the tests stated under Tablets.
Sulphated ash (2.3.18). Not more than 0.5 per cent. Assay. Determine by the microbiological assay of antibiotics,
Water (2.3.43). Not more than 4.0 per cent, determined on Method A (2.2.10) on a solution prepared in the following
0.3 g using a 10 per cent w/v solution of imidazole in manner.
anhydrous methanol as the solvent.
Assay. Determine by the microbiological assay of antibiotics, the powder containing about 25 mg of erythromycin and
Method A (2.2.10) using a solution prepared by dissolving dissolve as completely as possible in sufficient methanol to
476
IP 2007 ETHACRYNIC ACID TABLETS
produce 100.0 ml. Calculate the content of erythromycin in the dilute sulphuric acid and boil gently for 2 minutes; the
tablets, taking each 1000 Units found to be equivalent to 1 mg solution gives the reactions of chlorides (2.3.1).
of erythromycin.
Tests
Labelling. The label states the strength in terms of the Related substances. Determine by thin-layer chromatography
equivalent amount of erythromycin. (2.4.17), coating the plate with silica gel GF254.
volumes of ethyl acetate and 20 volumes of glacial acetic
acid.
Ethacrynic Acid Test solution. Dissolve 0.2 g of the substance under
Etacrynic Acid examination in 10 ml of ethanol (95 per cent).
O COOH substance under examination in ethanol (95 per cent).
CH2
H3C Reference solution (b). A 0.01 per cent w/v solution of the
Cl substance under examination in ethanol (95 per cent).
O Cl Apply to the plate 10 µl of each solution. After development.
C13H12Cl2O4 Mol. Wt. 303.1 Any secondary spot in the chromatogram obtained with the
Ethacrynic Acid is 2-[2,3-dichloro-4-(2-ethylacryloyl) test solution is not more intense than the spot in the
phenoxy]acetic acid chromatogram obtained with reference solution (a) and not
Ethacrynic Acid contains not less than 98.0 per cent and not
more than 102.0 per cent of C13H12Cl2O4, calculated on the
dried basis. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Description. A white or almost white, crystalline powder. Loss on drying (2.3.19). Not more than 0.5 per cent, determined
CAUTION - As Ethacrynic Acid irritates the skin, eyes and
60° at a pressure of 0.1 to 0.5 kPa.
the mucous membranes it should be handled with care.
Identification methanol and add 5 ml of water. Titrate with 0.1 M sodium
hydroxide solution, determining the end-point
potentiometrically (2.4.25).
C13H12Cl2O4.
Compare the spectrum with that obtained with ethacrynic
acid RS or with the reference spectrum of ethacrynic acid.
0.005 per cent w/v solution in a mixture of 99 volumes of
Ethacrynic Acid Tablets
methanol and 1 volume of 1 M hydrochloric acid shows a Etacrynic Acid Tablets
well defined absorption maximum at about 270 nm and a
Ethacrynic Acid Tablets contain not less than 90.0 per cent
shoulder at about 285 nm; absorbance at about 270 nm, 0.55 to
0.60.
ethacrynic acid, C13H12Cl2O4.
C. To 25 mg add 2 ml of 1 M sodium hydroxide and heat in a
water-bath for 5 minutes, cool, add 0.25 ml of sulphuric acid Identification
(50 per cent v/v) and 0.5 ml of a 10 per cent w/v solution of
Mix a quantity of the powdered tablets containing 50 mg of
chromotropic acid sodium salt and add cautiously 2 ml of
Ethacrynic Acid with 0.1 M hydrochloric acid and extract
sulphuric acid; a deep violet colour is produced.
with two quantities, each of 40 ml, of dichloromethane. Dry
D. On 20 mg determine by the oxygen-flask method (2.3.34), the combined extracts with anhrydrous sodium sulphate, filter
using 5 ml of dilute sodium hydroxide solution as the and evaporate to dryness with the aid of gentle heat. The
absorbing liquid. When the process is complete, acidify with residue complies with the following tests.
477
ETHAMBUTOL HYDROCHLORIDE IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). with a mixture of 70 volumes of acetonitrile and 30 volumes of
Compare the spectrum with that obtained with ethacrynic water.
acid RS or with the reference spectrum of ethacrynic acid. Chromatographic system
B. When examined in the range 230 nm to 360 nm (2.4.7), a – a stainless steel column 20 cm x 4 mm, packed with
0.005 per cent w/v solution in a mixture of 99 volumes of octadecylsilyl silica gel (10 µm),
methanol and 1 volume of 1 M hydrochloric acid shows a – mobile phase: a mixture of 60 volumes of water, 40
well defined absorption maximum at about 270 nm and a volumes of acetonitrile and 1 volume of glacial acetic
shoulder at about 285 nm; absorbance at about 270 nm, 0.55 to acid,
0.60. – flow rate. 2 ml per minute,
C. To 25 mg add 2 ml of 1 M sodium hydroxide and heat in a
water-bath for 5 minutes, cool, add 0.25 ml of sulphuric acid
(50 per cent v/v) and 0.5 ml of a 10 per cent w/v solution of Inject the test and reference solution.
chromotropic acid sodium salt and add cautiously 2 ml of
Calculate the content of C13H12Cl2O4 in the tablets.
sulphuric acid; a deep violet colour is produced.
Tests
Related substances. Determine by thin-layer chromatography Ethambutol Hydrochloride
Mobile phase. A mixture of 60 volumes of chloroform, 50 OH
volumes of ethyl acetate and 20 volumes of glacial acetic H
H3C N , 2HCl
acid. N CH3
H
OH
containing 0.2 g of Ethacrynic Acid with 10 ml of ethanol
(95 per cent) and filter. C10H24N2O2,2HCl Mol. Wt. 277.2
Reference solution (a). Dilute 3 volumes of the test solution Ethambutol Hydrochloride is (S,S)-N,N’-ethylenebis(2-
to 200 volumes with ethanol (95 per cent). aminobutan-1-ol) dihydrochloride.
Reference solution (b). Dilute 1 volume of the test solution to Ethambutol Hydrochloride contains not less than 97.0 per
200 volumes with ethanol (95 per cent). cent and not more than 101.0 per cent of C10H24N2O2,2HCl,
Apply to the plate 10 µl of each solution. After development, calculated on the dried basis.
dry the plate in air and examine in ultraviolet light at 254 nm. Description. A white, crystalline powder; almost odourless.
test solution is not more intense than the spot in the Identification
more than one such spot is more intense than the spot in the A. Determine by infrared absorption spectrophotometry (2.4.6).
chromatogram obtained with reference solution (b). Compare the spectrum with that obtained with ethambutol
Other tests. Comply with the tests stated under Tablets. ethambutol hydrochloride.
Assay. Determine by liquid chromatography (2.4.14). B. In the test for 2-Aminobutanol, the principal spot in the
Test solution (a). Weigh and powder 20 tablets. Shake a chromatogram obtained with test solution (b) corresponds to
quantity of the powdered tablets containing about 50 mg of that in the chromatogram obtained with reference solution
Ethacrynic Acid with 0.5 ml of glacial acetic acid and 50.0 ml (b).
of acetonitrile for 15 minutes and filter. C. A 5 per cent w/v solution gives reaction A of chlorides
Test solution (b). Prepare in the same manner as test solution (2.3.1).
(a) but using 0.5 ml of glacial acetic acid, 45 ml of acetonitrile Tests
and 5.0 ml of a 0.15 per cent w/v solution of propyl
hydroxybenzoate (internal standard) in acetonitrile. pH (2.4.24). 3.4 to 4.0, determined in a 2.0 per cent w/v solution.
Reference solution. Dissolve 50 mg of ethacrynic acid RS in Specific optical rotation (2.4.22). +6.0° to +6.6°, determined in
5.0 ml of the internal standard solution and dilute to 50.0 ml a 10.0 per cent w/v solution.
478
IP 2007 ETHAMBUTOL HYDROCHLORIDE
2-Aminobutanol. Determine by thin-layer chromatography – mobile phase: A. a mixture of equal volumes of methanol
(2.4.17), coating the plate with silica gel G. and water,
Mobile phase. A mixture of 75 volumes of methanol, 15 volumes B. water,
of water and 10 volumes of strong ammonia solution. – flow rate. 1 ml per minute,
Test solution (a). Dissolve 0.5 g of the substance under below,
examination in 10 ml of methanol. – spectrophotometer set at 215 nm,
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with – a 10 µl loop injector.
methanol. Time Mobile phase A Mobile phase B
Reference solution (a). A 0.05 per cent w/v solution of 2- (in min.) (per cent v/v) (per cent v/v)
aminobutanol RS in methanol. 0-30 82 18
Reference solution (b). A 0.5 per cent w/v solution of 31-35 0 100
ethambutol hydrochloride RS in methanol. 35-37 0 100
Apply to the plate 2 µl of each solution. After development, 37-38 82 18
dry the plate in air, heat at 110° for 10 minutes, cool, spray with
ninhydrin solution and heat at 110° for 5 minutes. Any spot
resolution between the peaks due to ethambutol and the RS
corresponding to 2-aminobutanol in the chromatogram
isomer is not less than 4.0 and the relative retention of RS
isomer with reference to ethambutol (retention time about 14
spot in the chromatogram obtained with reference solution (a).
min) is about 1.3.
Meso ethambutol (RS isomer). Determine by either of the
Inject reference solution (a) and the test solution.
following methods.
In the chromatogram obtained with the test solution, the area
Method A. Determine by differential scanning calorimetry
of the peak due to the RS isomer is not more than the area of
(DSC) (2.4.31).
the peak in the chromatogram obtained with reference solution
Test preparation: Weigh between 4 and 6 mg of the sample in (a) (1.0 per cent)
the 40 µl aluminium DSC crucible. Carry out the test by heating
at a rate of 10º per minute from 25º to 250º, under nitrogen
purging (20 ml/min) and record the thermogram. Observe the
endotherms at 42º ± 2º and 77º ± 2º corresponding to the Sulphated ash (2.3.18). Not more than 0.1 per cent.
transitions of the RS isomer and SS isomer, respectively. There
should not be any endothermic peak at 42º ± 2º in the
thermogram.
Method B. Determine by liquid chromatography (2.4.14)
Diluent. Dissolve 1.4 g of disodium hydrogen orthophosphate
Test solution. Suspend 4.0 mg of the substance under
anhydrous in 1000 ml of water and adjust the pH to 6.8 ± 0.05
examination in 4.0 ml of acetonitrile and 100 µl of triethylamine.
with orthophosphoric acid.
Stir the mixture with the aid of ultrasound for 5 minutes. Add
15 ìl of R-(+)-phenyl isocyanate and heat the mixture for 20 Test solution. Dissolve 30.0 mg of the substance under
minutes at 70º in a water-bath. examination in 100.0 ml of the diluent.
Reference solution (a). Dilute 1.0 ml of the test solution to Reference solution. A 0.03 per cent w/v solution of ethambutol
100.0 ml with acetonitrile. hydrochloride RS in the diluent.
Reference solution (b). Suspend 4.0 mg of ethambutol for Chromatographic system
system suitability RS (containing RS isomer) 4.0 ml of – a stainless steel column 15 cm x 4.6 mm, packed with
acetonitrile and 100 µl of triethylamine. Mix the mixture with nitrile groups chemically bonded to porous silica
the aid of ultrasound for 5 minutes. Add 15 ìl of R-(+)-phenyl particles (5 µm) ( such as Zorbax SB-CN),
isocyanate and heat the mixture for 20 minutes at 70º in a – mobile phase: a mixture of equal volumes of a buffer
water-bath. consisting of 1 ml of triethylamine in sufficient water
Chromatographic system to produce 1000 ml adjusted to pH 7.0 with
– a column 10 cm x 4.6 mm, packed with octadecylsilane orthophosphoric acid, and acetonitrile,
bonded to silica (3 µm) – flow rate. 1 ml per minute,
479
ETHAMBUTOL TABLETS IP 2007
– a 100 µl loop injector. Dissolution (2.5.2).

Inject the reference solution. The test is not valid unless the Apparatus. No 1
tailing factor is not more than 3.0 and the relative standard Medium. 900 ml of freshly distilled water
deviation for replicate injections is not more than 2.0 per cent. Speed and time. 100 rpm and 45 minutes.
Inject alternately the test solution and the reference solution. Withdraw a suitable volume of the medium and filter promptly
Calculate the content of C10H24N2O2,2HCl. through a membrane filter disc having an average pore diameter
Storage. Store protected from moisture. Dilute suitably with water to produce a solution containing
about 0.030 per cent w/v of ethambutol hydrochloride. Using
the resulting solution as the test solution carry out the
Ethambutol Tablets procedure described under Assay.
Ethambutol Hydrochloride Tablets Calculate the content of C10H24N2O2,2HCl.
Ethambutol Tablets contain not less than 95.0 per cent and D. Not less than 75 per cent of the stated amount of
not more than 105.0 per cent of the stated amount of C10H24N2O2,2HCl.
ethambutol hydrochloride, C10H24N2O2,2HCl. The tablets may Other tests. Comply with the tests stated under Tablets.
be coated.
Identification Prepare the following solutions freshly.
A. Extract a quantity of the powdered tablets containing 50 Test solution. Weigh and powder 20 tablets. Weigh accurately
mg of Ethambutol Hydrochloride with 5 ml of methanol, filter a quantity of the powder containing about 30 mg of Ethambutol
and evaporate the filtrate to dryness. The residue complies Hydrochloride, add 50 ml of water and shake for about 15
with the following test. minutes and add sufficient water to produce 100.0 ml. Filter
Determine by infrared absorption spectrophotometry (2.4.6). and discard the first 10 ml of the filtrate. Use the clear filtrate.
Compare the spectrum with that obtained with ethambutol Standard solution. A 0.03 per cent w/v solution of ethambutol
hydrochloride RS or with the reference spectrum of hydrochloride RS in water.
ethambutol hydrochloride.
B. Shake a quantity of the powdered tablets containing 0.1 g – a stainless steel column 15 cm x 4.6 mm, packed with
of Ethambutol Hydrochloride with 10 ml of water, filter, and to nitrile groups chemically bonded to porous silica
the filtrate add 2 ml of a 1 per cent w/v solution of copper particles (5 µm) ( such as Zorbax SB-CN),
sulphate and 1 ml of 1 M sodium hydroxide; a distinct blue – column. temperature 30°,
colour is produced. – mobile phase: a mixture of equal volumes of a buffer
consisting of 1 ml of triethylamine in sufficient water
Tests
to produce 1000 ml adjusted to pH 7.0 with
2-Aminobutanol. Determine by thin-layer chromatography orthophosphoric acid, and acetonitrile,
(2.4.17), coating the plate with silica gel G. – spectrophotometer set at 200 nm,
Mobile phase. A mixture of 75 volumes of methanol, 15 volumes – a 50 µl loop injector.
of water and 10 volumes of strong ammonia solution. Inject alternately the test solution and the reference solution.
Test solution. Shake a quantity of the powdered tablets Calculate the content of C10H24N2O2,2HCl in the tablets.
containing 0.5 g of Ethambutol Hydrochloride for 5 minutes
with sufficient methanol to produce 10 ml and filter.
Reference solution. A 0.05 per cent w/v solution of 2-
aminobutanol RS in methanol. Ethambutol and Isoniazid Tablets
dry the plate in air, heat at 110° for 5 minutes, cool, spray with Ethambutol Hydrochloride and Isoniazid Tablets
ninhydrin solution and heat at 110° for 5 minutes. Any spot Ethambutol and Isoniazid Tablets contain not less than
corresponding to 2-aminobutanol in the chromatogram 90.0 per cent and not more than 110.0 per cent of the stated
obtained with the test solution is not more intense than the amounts of ethambutol hydrochloride, C10H24N2O2.2HCl and
spot in the chromatogram obtained with the reference solution. isoniazid, C6H7N3O.
480
IP 2007 ETHAMBUTOL AND ISONIAZID TABLETS
Identification 263 nm (2.4.7). Calculate the content of C6H7N3O in the medium

from the absorbance obtained by repeating the determination
A. In the Assay, the principal peak in the chromatogram using a 0.0015 per cent w/v solution of isoniazid RS in place
obtained with the test solution corresponds to the peak due of the filtrate.
to ethambutol hydrochloride RS in the chromatogram
obtained with the reference solution. D. Not less than 75 per cent of the stated amount of
C10H24N2O2.2HCl and C6H7N3O.
to isoniazid RS in the chromatogram obtained with the Assay. For isoniazid - Determine by liquid chromatography
reference solution. (2.4.14).
Diluent. Dissolve 1.4 g of disodium hydrogen orthophosphate
Tests anhydrous in water, adjust the pH to 6.8 ± 0.05 with dilute
2-Aminobutanol. Determine by thin-layer chromatography phosphoric acid and add sufficient water to produce 1000 ml.
(2.4.17), coating the plate with silica gel G. Test solution. Weigh and powder 20 tablets. Weigh accurately
a quantity of powdered tablets containing about 40 mg of
Mobile phase. A mixture of 75 volumes of methanol, 15 volumes
Isoniazid, dissolve in 50.0 ml of methanol and dilute to
of water and 10 volumes of strong ammonia solution.
500.0 ml with the diluent.
Reference solution. Weigh accurately about 40 mg of isoniazid
containing 0.5 g of ethambutol hydrochloride for 5 minutes
RS, dissolve in 50.0 ml of methanol and dilute to 500.0 ml with
with sufficient methanol to produce 10 ml and filter.
the diluent.
Reference solution. A 0.05 per cent w/v solution of 2- Chromatographic system
aminobutanol RS in methanol. – a stainless steel column 15 cm x 4.6 mm, packed with
Apply to the plate 2 µl of each solution. After development, octadecylsilyl silica gel (5 µm) (such as Intersil ODS-3),
dry the plate in air, heat at 110° for 5 minutes, cool, spray with – column temperature 30°,
ninhydrin solution and heat at 110° for 5 minutes. Any spot – mobile phase: 96 volumes of buffer solution pH 6.8
corresponding to 2-aminobutanol in the chromatogram prepared by dissolving 1.4 g disodium hydrogen
obtained with the test solution is not more intense than the orthophsophate anhydrous in 1000 ml of water, the pH
spot in the chromatogram obtained with the reference solution. of which is adjusted to 6.8 ± 0.05 with dilute phosphoric
Dissolution (2.5.2). acid and 4 volumes of acetonitrile,
Apparatus. No 1 – spectrophotometer set at 254 nm,
Medium. 900 ml of water – a 20 µl loop injector.
Withdraw a suitable volume of the medium and filter promptly tailing factor is not more than 2.0, the column efficiency
through a membrane filter disc having an average pore diameter determined from the isoniazid peak is not more than 1500
not greater than 1.0 µm, rejecting the first 10 ml of the filtrate. theoretical plates and the relative standard deviation for
On the filtrate determine by liquid chromatography (2.4.14). replicate injections is not more than 2.0 per cent.
For Ethambutol Hydrochloride —
Calculate the content of C6H7N3O in the tablets.
Test solution. Dilute the filtrate to obtain 0.044 per cent w/v
solution in the dissolution medium. For ethambutol hydrochloride -— Determine by liquid
ethambutol hydrochloride RS in the dissolution medium. Test solution. Weigh and powder 20 tablets. Weigh accurately
a quantity of the powder containing about 60 mg of Ethambutol
Determine the content of C10H24N2O2.2HCl by the procedure Hydrochloride and dissolve in 100.0 ml of the diluent.
given under Assay of Ethambutol hydrochloride.
Reference solution. A 0.06 per cent w/v solution of ethambutol
Calculate the content of C10H24N2O2.2HCl in the medium. hydrochloride RS in the diluent.
For Isoniazid — Determine the amount of C6H7N3O dissolved Chromatographic system
by measuring the absorbance of the filtrate, suitably diluted – a stainless steel column 15 cm x 4.6 mm, packed with
with the dissolution medium to obtain a solution containing nitrile groups chemically bonded to porous silica
about 0.015 mg of isoniazid per ml, at the maximum at about particles (such as Zorbax SB CN 5 µm),
481
ETHANOL IP 2007
– mobile phase: a mixture of 50 volumes of buffer pH 7.0 Acidity or alkalinity. To 20 ml add 0.25 ml of phenolphthalein
prepared by mixing 1 ml of triethylamine in 1000 ml of solution; the solution remains colourless and requires not
water the pH of which is adjusted to 7.0 ± 0.05 with more than 0.2 ml of 0.1 M sodium hydroxide to produce a pink
phosphoric acid and 50 volumes of acetonitrile. colour.
– flow rate. 1 ml per minute, Methanol. To 1 drop add 1 drop of water, 1 drop of dilute
– spectrophotometer set at 200 nm, phosphoric acid and 1 drop of potassium permanganate
– a 50 µl loop injector. solution. Mix, allow to stand for 1 minute and add sodium
Inject the reference solution. The test is not valid unless the bisulphite solution dropwise until the permanganate colour
tailing factor is not more than 3.0, the column efficiency is discharged. If a brown colour remains, add 1 drop of dilute
determined from Isoniazid peak is not more than 1500 theoretical phosphoric acid. To the colourless solution add 5 ml of freshly
plates and the relative standard deviation for replicate prepared chromotropic acid solution and heat on a water-
injections is not more than 2.0 per cent. bath at 60° for 10 minutes; no violet colour is produced.
Inject alternately the test solution and reference solution. Foreign organic substances. Clean a glass-stoppered cylinder
thoroughly with hydrochloric acid, rinse with water and finally
Calculate the content of C10H24N2O2.2HCl the tablets.
rinse with the substance under examination. Put 20 ml in the
Storage. Store protected from moisture. cylinder, cool to about 15° and then add from a carefully
cleaned pipette 0.1 ml of 0.1 M potassium permanganate. Mix
at once by inverting the stoppered cylinder and allow to stand
at 15° for 5 minutes; the pink colour does not entirely disappear.
Ethanol
2-Propanol and 2-methyl-2-propanol. To 1 ml add 3 ml of water
Absolute Alcohol; Dehydrated Alcohol and 10 ml of mercuric sulphate solution and heat in a boiling
CH3CH2OH Mol. Wt. 46.1 water-bath; no precipitate is formed within 3 minutes.
Ethanol contains not less than 99.0 per cent w/w and not more Aldehydes. Not more than 10 ppm, determined by the following
than 100.0 per cent w/w, corresponding to not less than method. To 5.0 ml add 5 ml of water and 1 ml of decolorised
99.4 per cent v/v and not more than 100.0 per cent v/v, at magenta solution and allow to stand for 30 minutes. Any
15.56°, of C2H6O. colour produced is not more intense than that produced by
treating in the same manner 5.0 ml of a 0.001 per cent w/v
Description. A clear, colourless, mobile and volatile liquid; solution of redistilled acetaldehyde in aldehyde-free ethanol
odour, characteristic and spirituous; hygroscopic. Readily (95 per cent).
volatilises even at low temperature; boils at 78°; flammable,
burning with a blue, smokeless flame. Benzene and related substances. Determine by gas
Identification Test solution. The substance under examination.
A. Mix 0.25 ml in a small beaker with 1 ml of potassium Reference solution (a). A 0.1 per cent v/v solution of
permanganate solution and 0.25 ml of dilute sulphuric acid 2-butanol reagent in the test solution.
and cover the beaker immediately with a filter paper moistened Reference solution (b). A solution containing 0.1 per cent v/v
with a solution freshly prepared by dissolving 0.1 g of sodium each of 2-butanol reagent and 1-propanol in the test solution.
nitroprusside and 0.5 g of piperazine hydrate in 5 ml of water;
Reference solution (c). A 0.0002 per cent v/v solution of
an intense blue colour is produced on the filter paper, the
benzene in the test solution.
colour becoming lighter after a few minutes.
B. To 5 ml of a 0.5 per cent v/v solution add 1 ml of 1 M sodium
– a glass column 1.8 m x 2 mm, packed with acid-washed
hydroxide followed by slow addition of 2 ml of iodine solution;
diatomaceous support (80 to 100 mesh) coated with 15
the odour of iodoform develops and a yellow precipitate is
per cent w/w of polyethyelene glycol 400,
produced.
– temperature:
Tests column. 50°,
inlet port. 150°,
Relative density (2.4.29). 0.7871 to 0.7902, determined at 25°. – flame ionisation detector at 250°,
Appearance of solution. Dilute 5.0 ml to 100.0 ml with water. – flow rate. 30 ml per minute of the carrier gas (nitrogen).
The solution is clear (2.4.1). Cool to 10° for 30 minutes; the Inject separately 2 µl of each of the test solution and reference
solution remains clear. solution (a). The chromatogram obtained with the test solution
482
IP 2007 ETHANOL (95 PER CENT)
shows no peak with a retention time similar to the peak due to Identification
2-butanol (retention time relative to isopropyl alcohol, about
1.5) obtained with reference solution (a). Inject 2 µl of reference A. Mix 0.25 ml in a small beaker with 1 ml of potassium
solution (b) and adjust the sensitivity of the system so that permanganate solution and 0.25 ml of dilute sulphuric acid
the heights of the peaks due to 2-butanol and propanol in the and cover the beaker immediately with a filter paper moistened
chromatogram obtained with reference solution (b) are not with a solution freshly prepared by dissolving 0.1 g of sodium
less than 50 per cent of the full scale of the recorder. nitroprusside and 0.5 g of piperazine hydrate in 5 ml of water;
an intense blue colour is produced on the filter paper, the
The test is not valid unless the resolution between the peaks colour becoming lighter after a few minutes.
due to 2-butanol and propanol in the chromatogram obtained
with reference solution (b) is at least 1.2. B. To 5 ml of a 0.5 per cent v/v solution add 1 ml of 1 M sodium
hydroxide followed by slow addition of 2 ml of iodine solution;
Inject alternately 2 µl each of the test solution and reference the odour of iodoform develops and a yellow precipitate is
solution (c). The area of any peak due to benzene in the produced.
chromatogram obtained with the test solution is not greater
than the difference between the area of the peak due to benzene Tests
in the chromatogram obtained with reference solution (c) and
Relative density (2.4.29). 0.8084 to 0.8104, determined at 25°.
that of the peak due to benzene in the chromatogram obtained
with the test solution. Appearance of solution. Dilute 5.0 ml to 100.0 ml with water.
The solution is clear (2.4.1). Cool to 10° for 30 minutes; the
In the chromatogram obtained with reference solution (a) the
solution remains clear.
sum of the areas of any peaks other than the principal peak
and the peaks due to 2-butanol is not greater than 3 times the Acidity or alkalinity. To 20 ml add 0.25 ml of phenolphthalein
area of the peak due to 2-butanol (0.3 per cent). solution; the solution remains colourless and requires not
more than 0.2 ml of 0.1 M sodium hydroxide to produce a pink
Fusel oil constituents. Place 25 ml in a porcelain dish protected
colour.
from dust and allow the liquid to evaporate on a water-bath
until a little of the liquid remains. Remove the dish from the Methanol. To 1 drop add 1 drop of water, 1 drop of dilute
water-bath and allow the liquid to evaporate at room phosphoric acid and 1 drop of potassium permanganate
temperature till the dish is almost dry. No foreign odour is solution. Mix, allow to stand for 1 minute and add sodium
perceptible. Add 1 ml of sulphuric acid; no red or brown bisulphite solution dropwise until the permanganate colour
colour is produced. is discharged. If a brown colour remains, add 1 drop of dilute
phosphoric acid. To the colourless solution add 5 ml of freshly
Non-volatile matter. Evaporate 100.0 ml in a tared dish on a prepared chromotropic acid solution and heat on a water-
water-bath and dry the residue at 105°; the residue weighs not bath at 60° for 10 minutes; no violet colour is produced.
more than 5 mg.
Foreign organic substances. Clean a glass-stoppered cylinder
Storage. Store in tightly-closed containers at a temperature thoroughly with hydrochloric acid, rinse with water and finally
not exceeding 30°, away from fire and protected from moisture. rinse with the substance under examination. Put 20 ml in the
Labelling. The label states that it is flammable. cylinder, cool to about 15° and then add from a carefully
cleaned pipette 0.1 ml of 0.1 M potassium permanganate. Mix
at once by inverting the stoppered cylinder and allow to stand
at 15° for 5 minutes; the pink colour does not entirely disappear.
Ethanol (95 Per Cent) 2-Propanol and 2-methyl-2-propanol. To 1 ml add 3 ml of water
and 10 ml of mercuric sulphate solution and heat in a boiling
Alcohol (95 per cent)
water-bath; no precipitate is formed within 3 minutes.
Ethanol (95 per cent) is a mixture of Ethanol and Water.
Aldehydes. Not more than 10 ppm, determined by the following
Ethanol (95 per cent) contains not less than 92.0 per cent w/w method. To 5.0 ml add 5 ml of water and 1 ml of decolorised
and not more than 92.7 per cent w/w, corresponding to not magenta solution and allow to stand for 30 minutes. Any
less than 94.7 per cent v/v and not more than 95.2 per cent colour produced is not more intense than that produced by
v/v, at 15.56°, of C2H6O. treating in the same manner 5 ml of a 0.001 per cent w/v solution
Description. A clear, colourless, mobile and volatile liquid; of redistilled acetaldehyde in aldehyde-free ethanol (95 per
odour, characteristic and spirituous. It is readily volatilised cent).
even at low temperatures; boils at about 78°; flammable, Benzene and related substances. Determine by gas
burning with a blue, smokeless flame. chromatography (2.4.13).
483
ANAESTHETIC ETHER IP 2007
Test solution. The substance under examination. Storage. Store in tightly-closed containers at a temperature
Reference solution (a). A 0.1 per cent v/v solution of 2- not exceeding 30° and away from fire.
butanol reagent in the test solution. Labelling. The label states that it is flammable.
v/v each of 2-butanol reagent and 1-propanol in the test
solution. Anaesthetic Ether
Reference solution (c). A 0.0002 per cent v/v solution of CH3CH2OCH2CH3
benzene in the test solution.
C4H10O Mol. Wt. 74.1
– a glass column 1.8 m x 2 mm, packed with acid-washed Anaesthetic Ether is diethyl ether to which a suitable non-
diatomaceous support (80 to 100 mesh) coated with 15 volatile stabiliser in a proportion not greater than 0.002 per
per cent w/w of polyethyelene glycol 400, cent w/v may have been added.
– temperature: Description. A clear, colourless, very mobile liquid; odour,
column. 50°, characteristic; highly flammable.
inlet port. 150°,
– flame ionisation detector at 250°, NOTE — It is absolutely essential that a preservative of the
– flow rate. 30 ml per minute of the carrier gas (nitrogen). type of sodium pyrogallate, hydroquinone or propyl gallate
in suitable concentrations shall be added in Anaesthetic
Inject alternately 2 µl of each of the test solution and reference
Ether intended for use in tropical climates unless the
solution (a). The chromatogram obtained with the test solution
Anaesthetic Ether is stored in a copper container or in a
shows no peak with a retention time similar to the peak due to
container copper-plated internally. The preservative used
2-butanol (retention time relative to isopropyl alcohol, about
and its concentration shall be declared on the label.
1.5) obtained with reference solution (a). Inject 2 µl of reference
solution (b) and adjust the sensitivity of the system so that Tests
the heights of the peaks due to 2-butanol and propanol in the
chromatogram obtained with reference solution (b) are not Relative density (2.4.29). 0.714 to 0.716, determined at 20°.
less than 50 per cent of the full scale of the recorder. Boiling range (2.4.8). 34° to 35°.
The test is not valid unless the resolution between the peaks CAUTION — It is dangerous to determine the boiling range
due to 2-butanol and propanol in the chromatogram obtained if the sample does not comply with the test for peroxides.
with reference solution (b) is at least 1.2. Acidity. To 20 ml of ethanol (95 per cent) add 0.25 ml of
Inject alternately 2 µl each of the test solution and reference bromothymol blue solution add dropwise 0.02 M sodium
solution (c). The area of any peak due to benzene in the hydroxide until the blue colour persists for 30 seconds. Add
chromatogram obtained with the test solution is not greater 25 ml of the substance under examination, shake and again
than the difference between the area of the peak due to benzene add dropwise 0.2 M sodium hydroxide until the blue colour
in the chromatogram obtained with reference solution (c) and reappears and persists for 30 seconds. Not more than 0.4 ml of
that of the peak due to benzene in the chromatogram obtained 0.02 M sodium hydroxide is required.
with the test solution. Peroxides. Place 8 ml of potassium iodide and starch solution
In the chromatogram obtained with reference solution (a) the in a 12-ml glass-stoppered cylinder of about 1.5 cm diameter.
sum of the areas of any peaks other than the principal peak Fill completely with the substance under examination, insert
and the peaks due to 2-butanol is not greater than 3 times the the stopper, shake vigorously and allow to stand in the dark
area of the peak due to 2-butanol (0.3 per cent). for 30 minutes; no colouration is produced.
Fusel oil constituents. Place 25 ml in a porcelain dish protected Acetone and aldehydes. Place 2 ml of alkaline potassium
from dust and allow the liquid to evaporate on a water-bath mercuri-iodide solution in a 12-ml glass-stoppered cylinder
until a little of the liquid remains. Remove the dish from the of about 1.5 cm diameter and fill completely with the substance
water-bath and allow the liquid to evaporate at room under examination, insert the stopper and shake vigorously
temperature till the dish is almost dry. No foreign odour is for 15 seconds and set aside for 5 minutes, protected from
perceptible. Add 1 ml of sulphuric acid; no red or brown light; no colour or turbidity, except for slight opalescence, is
colour is produced. produced.
Non-volatile matter. Evaporate 100.0 ml in a tared dish on a If the ether does not comply with the test, distil 40 ml (after
water-bath and dry the residue at 105°; the residue weighs not ensuring that it complies with the test for peroxides) until
more than 5 mg. only 5 ml remains and repeat the test using 10 ml of the distillate.
484
IP 2007 ETHINYLOESTRADIOL
Foreign odour. Pour 10 ml in successive portions on to a clean ethinyloestradiol RS or with the reference spectrum of
filter paper and allow to evaporate spontaneously; no foreign ethinyloestradiol.
odour is detectable at any stage of evaporation. B. In the test for Related substances, the principal spot in the
Non-volatile matter. Evaporate 50 ml in a tared dish on a water- chromatogram obtained with test solution (b) corresponds to
bath and dry at 105°(after ensuring that the sample complies that in the chromatogram obtained with reference solution
with the test for peroxides); the residue weighs not more than (b).
1.5 mg.
C. Dissolve about 1 mg in 1 ml of sulphuric acid; an orange-
Methanol. To 10 ml, add 5 ml of ethanol (20 per cent) and 5 ml red colour develops which exhibits a greenish fluorescence
of water, in a separator, shake vigorously, set aside and allow when examined in ultraviolet light at 365 nm. Add the solution
the mixture to separate and draw off the lower layer. To 5 ml of to 10 ml of water; the colour changes to violet and a violet
the lower layer add 2.0 ml of potassium permanganate and precipitate is produced.
phosphoric acid solution, set aside for 10 minutes and add
2.0 ml of oxalic acid and sulphuric acid solution and 5 ml of Tests
decolorised magenta solution. Set aside for 30 minutes; no
colour is produced. Appearance of solution. A 5.0 per cent w/v solution in ethanol
is clear (2.4.1), and not more intensely coloured than reference
Water (2.3.43). Not more than 0.2 per cent, determined on solution BYS6 (2.4.1).
20.0 ml.
Storage. Store protected from light at a temperature not at 20° in a 5.0 per cent w/v solution in pyridine.
exceeding 30°. Ether remaining in a partly used container may
deteriorate rapidly. Light absorption (2.4.7). Absorbance of a 0.01 per cent w/v
solution in ethanol (95 per cent) at about 281 nm, 0.69 to 0.73.
Labelling. The label states that (1) it is very flammable and
should not be used near a naked flame; (2) the name and Related substances. Determine by thin-layer chromatography
proportion of any stabiliser added. (2.4.17), coating the plate with silica gel G..
Mobile phase. A mixture of 90 volumes of toluene and 10
volumes of ethanol (95 per cent).
Ethinyloestradiol Test solution (a). Dissolve 0.2 g of the substance under
Ethinylestradiol examination in 10 ml of a mixture of 9 volumes of chloroform
and 1 volume of methanol.
H3C OH Test solution (b). Dilute 5 ml of test solution (a) to 100 ml with
C CH
H
H H with the same solvent mixture.
HO Reference solution (b). A 0.1 per cent w/v solution of
ethinyloestradiol RS in the same solvent mixture.
C20H24O2 Mol. Wt. 296.4
Reference solution (c). A 0.02 per cent w/v solution of estrone
Ethinylestradiol is 19-nor-17α-pregna-1,3,5(10)-trien-20yne-
3,17β-diol.
Ethinyloestradiol contains not less than 97.0 per cent and not Apply to the plate 5 µl of each solution. After development,
more than 102.0 per cent of C20H24O2, calculated on the dried dry the plate in air until the odour of the solvent is no longer
basis. detectable, heat at 110° for 10 minutes and spray the hot plate
with ethanolic sulphuric acid (20 per cent v/v). Heat again at
Description. A white or slightly yellowish-white, crystalline 110° for 10 minutes and examine in ultraviolet light at 365 nm.
powder. In the chromatogram obtained with test solution (a) any spot
corresponding to estrone is not more intense than the spot in
Identification
the chromatogram obtained with reference solution (c) and
Test A may be omitted if tests B and C are carried out. Test C any other secondary spot is not more intense than the spot in
may be omitted if tests A and B are carried out. the chromatogram obtained with reference solution (a).
A. Determine by infrared absorption spectrophotometry (2.4.6). Loss on drying (2.4.19). Not more than 1.0 per cent, determined
Compare the spectrum with that obtained with on 0.5 g by drying in an oven at 105° for 3 hours.
485
ETHINYLOESTRADIOL TABLETS IP 2007
Assay. Dissolve 0.2 g in 40 ml of tetrahydrofuran, add 5 ml of and filter. Take 1 ml of this solution dilute 10 ml with the same
a 10 per cent w/v solution of silver nitrate and titrate with 0.1 solvent.
M sodium hydroxide, determining the end-point Reference solution. A 0.0025 per cent w/v solution of
potentiometrically (2.4.25). Carry out a blank titration. ethinylestradiol RS in mobile phase.
C20H24O2.
Storage. Store protected from light. – mobile phase: a mixture of 60 volume of acetonitrile
and 40 volumes of water.
Ethinyloestradiol Tablets – spectrophotometer set at 280 nm,
Ethinylestradiol Tablets – a 20 µl loop injector.
Ethinyloestradiol Tablets contain not less than 90.0 per cent Inject the test solution and reference solution.
and not more than 110.0 per cent of the stated amount of Calculate the content of C20H24O2 in the tablet.
ethinyloestradiol, C20H24O2.
Identification Assay. Determine by liquid chromatography (2.4.14).
A. Determine by thin-layer chromatography (2.4.17), coating Test solution. Weigh and powder 20 tablets. Weight accurately
the plate with silica gel G. a quantity of the powder containing 2.5 mg of ethinylestradiol,
Mobile phase. A mixture of 90 volumes of toluene and 10 add 20.0 ml of mobile phase, shake for 10 minutes, dilute to
volumes of ethanol (95 per cent). 100 ml and filter. Take 1 ml of this solution and dilute to 10 ml
containing 0.25 mg of Ethinyloestradiol with four quantities, Reference solution. A 0.0025 per cent w/v solution of
each of 20 ml of chloroform, filter each extract in turn, evaporate ethinylestradiol RS in mobile phase.
the combined filtrates to dryness on a water-bath in a current Carry out the chromatographic procedure described under
of nitrogen and dissolve the residue in 0.25 ml of chloroform. Uniformity of content. Calculate the content of C20H24O2 in
Reference solution. A 0.1 per cent w/v solution of the tablets.
ethinyloestradiol RS in chloroform. Storage. Store protected from light.
dry the plate in air, spray with ethanolic sulphuric acid
(20 per cent v/v), heat at 110° for 10 minutes and examine in
ultraviolet light at 365 nm and in daylight. By both methods of
Ethionamide
visualisation, the principal spot in the chromatogram obtained N
with the test solution corresponds to that in the chromatogram CH3
B. Triturate a quantity of the powdered tablets containing 0.1
mg of Ethinyloestradiol with 0.5 ml of 0.1 M sodium hydroxide S NH2
and 5 ml of water, allow to stand for 5 minutes, filter, acidify
the filtrate with 0.15 ml of sulphuric acid, add 3 ml of ether, C8H10N2S Mol. Wt. 166.2
shake and allow to separate. Evaporate the ether layer to Ethionamide is 2-ethylpyridine-4-carbothioamide.
dryness and heat the residue on a water-bath for 5 minutes
with 0.2 ml of glacial acetic acid and 2 ml of phosphoric acid; Ethionamide contains not less than 98.5 per cent and not more
a pink colour with an intense orange fluorescence is produced. than 101.0 per cent of C8H10N2S, calculated on the dried basis.
Tests Description. A yellow crystalline powder or small yellow

crystals.
Tablets. Identification
Determine by liquid chromatography (2.4.14). A. Determine by infrared absorption spectrophotometry (2.4.6).
Test solution. Finely crush one tablet, add 20.0 ml of mobile Compare the spectrum with that obtained with ethionamide
phase, shake for 10 minutes, dilute to 100 ml with mobile phase RS or with the reference spectrum of ethionamide.
486
IP 2007 ETHIONAMIDE TABLETS
B. Dissolve about 10 mg in 5 ml of methanol and add 5 ml of Test solution. Dissolve about 50 mg of the substance under
0.1 M silver nitrate; a dark brown precipitate is produced. examination in 100.0 ml of the mobile phase. Dilute 5.0 ml of
C. Melting point (2.4.21). 158° to 164°. this solution to 50.0 ml with the mobile phase.
Reference solution. Dissolve 50 mg of the ethionamide RS in
Tests 100.0 ml of the mobile phase. Dilute 5.0 ml of this solution to
Appearance of solution. Dissolve 0.5 g in 10 ml of methanol,
heating to about 50° and allow to cool to room temperature. Inject the reference solution. The test is not valid unless the
The solution is not more opalescent than opalescence standard relative standard deviation for replicate injections is not more
OS2 (2.4.1). than 2.0 per cent, the tailing factor is not more than 2.0 and the
column efficiency in not less than 5000 theoretical plates.
Acidity. Dissolve 2.0 g in 20 ml of methanol, heating to about
50°, and add 20 ml of water. Cool slightly, shake until Inject alternately the test solution and the reference solution.
crystallisation occurs and allow to cool to room temperature. Calculate the content of C8H10N2S.
Add 60 ml of water and titrate with 0.1 M sodium hydroxide
using 0.2 ml of cresol red solution as indicator. Not more than
0.2 ml is required to change the colour of the solution to red.
(2.4.14).
Ethionamide Tablets
Test solution. Dissolve 50 mg of the substance under Ethionamide Tablets contain not less than 95.0 per cent and
examination in 100 ml of the mobile phase. not more than 105.0 per cent of ethionamide, C8H10N2S. The
tablets may be coated.
100 ml of the mobile phase. Dilute 1 ml of this solution to100 ml Identification
A. Extract a quantity of the powdered tablets containing 25
Chromatographic system mg of Ethionamide with 5 ml of methanol, filter and evaporate
– a stainless steel column 25 cm x 4.6 mm, packed with the filtrate to dryness. The residue complies with the following
octadecylsilyl silica gel (5 µm) (such as Intersil ODS-3), test.
– mobile phase: a mixture of 60 volumes of a buffer
prepared by dissolving 2 ml of triethylamine in water,
Compare the spectrum with that obtained with ethionamide
adjusting the pH to 6.0 with orthophosphoric acid and
RS or with the reference spectrum of ethionamide.
diluting to 1000 ml, and 40 volumes of acetonitrile and
filtered, B. In the Assay, the principal peak in the chromatogram
– flow rate. 1 ml per minute, obtained with the test solution corresponds the peak in the
– spectrophotometer set at 290 nm, chromatogram obtained with the reference solution.
Tests
(2.4.14) as given under Assay using the following solutions.
than 2.0 per cent.
Test solution. Weigh accurately a quantity of the powder
Inject the test solution and the reference solution. Any
containing 50 mg of Ethionamide and dissolve in 100 ml of the
individual impurity is not more than 0.5 per cent and the sum
mobile phase.
of all impurities found is not more than 1.0 per cent.
100 ml of the mobile phase. Dilute 1 ml of this solution to100 ml
Loss on drying (2.4.19). Not more than 0.5 per cent, determined relative standard deviation for replicate injections is not more
on 1.0 g by drying in an oven at 105° for 3 hours. than 2.0 per cent.
Assay. Determine by liquid chromatography (2.4.14) as given Inject the test solution. Any individual impurity is not more
under the test for Related substances using the following than 0.5 per cent and the sum of all impurities found is not
solutions. more than 1.0 per cent.
487
ETHOPROPAZINE HYDROCHLORIDE IP 2007
Dissolution (2.5.2). Ethoprozanine Hydrochloride is 10-[2-

Apparatus. No 2 (diethylamino)propyl]phenothiazine hydrochloride.
Medium. 900 ml of 0.1 M hydrochloric acid Ethopropazine Hydrochloride contains not less than 99.0 per
Speed and time. 100 rpm and 45 minutes. cent and not more than 101.5 per cent of C19H24N2S,HCl,
Withdraw a suitable volume of the medium, filter and dilute a calculated on the dried basis.
suitable volume of the filtrate with the same solvent. Measure Description. A white or slightly creamy-white, crystalline
the absorbance of the resulting solution at the maximum at powder.
about 274 nm (2.7.4). Calculate the content of C8H10N2S from
the absorbance of a solution of known concentration of Identification
ethionamide RS.
D. Not less than 75 per cent of the stated amount of C8H10N2S. Compare the spectrum with that obtained with ethopropazine
Other tests. Comply with the tests stated under Tablets. hydrochloride RS.
Assay. Determine by liquid chromatography (2.4.14). B. When examined in the range 230 nm to 360 nm (2.4.7), a
Test solution. Weigh and powder 20 tablets. Weigh accurately 0.0005 per cent w/v solution in ethanol (95 per cent) shows
a quantity of the powder containing about 50 mg of Ethionamide an absorption maximum at about 252 nm and a less well-defined
in 100.0 ml of the mobile phase. Dilute 5.0 ml of this solution to maximum at about 303 nm; absorbance at about 252 nm, about
50.0 ml with the mobile phase. 0.42.
Reference solution. Dissolve 50 mg of the ethionamide RS in C. In the test for Related substances, the principal spot in the
100 ml of the mobile phase. Dilute 5.0 ml of this solution to 50.0 chromatogram obtained with the test solution corresponds to
ml with the mobile phase. that in the chromatogram obtained with reference solution
(b).
– a stainless steel column 25 cm x 4.6 mm, packed with D. Gives reaction A of chlorides (2.3.1).
octadecylsilyl silica gel (5 µm) (such as Intersil ODS-3),
– mobile phase: a mixture of 40 volumes of acetonitrile Tests
and 60 volumes of buffer pH 6.0 prepared by mixing 2 ml Acidity or alkalinity. Dissolve 0.15 g in 50 ml of carbon
of triethylamine to 1000 ml with water and adjusting dioxide-free water and add 0.15 ml of methyl red solution; the
the pH to 6.0 with phosphoric acid, solution is yellow and not more than 0.2 ml of 0.01 M
– flow rate. 1 ml per minute, hydrochloric acid is required to change the colour of the
– spectrophotometer set at 290 nm, solution to red.
than 2.0 per cent, the tailing factor is not more than 2.0 and the Mobile phase. A freshly prepared mixture of equal volumes of
column efficiency in not less than 5000 theoretical plates. ether and ethyl acetate saturated with strong ammonia
Inject alternately the test solution and the reference solution. solution.
Calculate the content of C8H10N2S in the tablets. Test solution. Dissolve 0.5 g of the substance under
Storage. Store protected from light and moisture. examination in 100 ml of methanol.
100 volumes with methanol.
Ethopropazine Hydrochloride Reference solution (b). A 0.5 per cent w/v solution of
ethopropazine hydrochloride RS in methanol.
H3C N CH3 Apply to the plate 2 µl of each solution. After development,
CH3
N test solution is not more intense than the spot in the
,HCl
S Heavy metals (2.3.13). 1.0 g complies with the limit test for
C19H24N2S,HCl Mol. Wt. 348.9 heavy metals, Method B (20 ppm).
488
IP 2007 ETHOSUXIMIDE
Sulphated ash (2.3.18). Not more than 0.1 per cent. Reference solution (a). A 0.2 per cent w/v solution of
Loss on drying (2.4.19). Not more than 0.5 per cent, determined ethopropazine hydrochloride RS in chloroform.
on 1.0 g by drying in an oven at 105°. Reference solution (b). A 0.002 per cent w/v solution of
ethopropazine hydrochloride RS in chloroform.
acetone, add 15 ml of mercuric acetate solution. Titrate with Apply to the plate 2 µl of each solution. After development,
0.1 M perchloric acid, using 0.15 ml of a saturated solution of dry the plate in air and examine in ultraviolet light at 254 nm.
methyl orange in acetone as indicator. Carry out a blank Any secondary spot in the chromatogram obtained with the
titration. test solution is not more intense than the spot in the
C19H24N2S,HCl. Other tests. Comply with the tests stated under Tablets.
Storage. Store protected from light. Assay. Protect the solution from light throughout the test.
the powder containing about 50 mg of Ethopropazine
Hydrochloride, extract with four quantities, each of 20 ml, of
Ethopropazine Tablets ethanol (95 per cent). Filter and dilute the filtrate to 100.0 ml
with ethanol (95 per cent). Dilute 10.0 ml of this solution to
Ethopropazine Hydrochloride Tablets
100.0 ml with ethanol (95 per cent). Dilute 10.0 ml of this
Ethopropazine Tablets contain not less than 92.5 per cent and solution further to 100.0 ml and measure the absorbance of
not more than 107.5 per cent of the stated amount of the resulting solution at the maximum at about 252 nm (2.4.7).
ethopropazine hydrochloride, C19H24N2S,HCl. The tablets may Calculate the content of C19H24N2S,HCl, taking 845 as the
be coated. specific absorbance at 252 nm.
Identification Storage. Store protected from light.
A. Extract a quantity of the powdered tablets containing 50

mg of Ethopropazine Hydrochloride with 20 ml of chloroform,
filter, evaporate the filtrate to dryness and dry the residue at Ethosuximide
60° at a pressure not exceeding 0.7 kPa. The residue complies
H
Determine by infrared absorption spectrophotometry (2.4.6). O N O
Compare the spectrum with that obtained with ethopropazine
CH3
hydrochloride RS.
CH3
that in the chromatogram obtained with reference solution (a). C7H11NO2 Mol. Wt. 141.2
C. To a quantity of the powdered tablets containing 5 mg of Ethosuximide is (RS)-2-ethyl-2-methylsuccinimide.

Ethopropazine Hydrochloride add 5 ml of sulphuric acid and Ethosuximide contains not less than 98.0 per cent and not
allow to stand for 5 minutes; a red colour is produced. more than 101.0 per cent of C7H11NO2, calculated on the
anhydrous basis.
Tests
Description. A white or almost white powder or waxy solid.
(2.4.17), coating the plate with silica gel G. Identification
Mobile phase. A freshly prepared mixture of equal volumes of Test A may be omitted if tests B and C are carried out. Tests B
ether and ethyl acetate saturated with strong ammonia and C may be omitted if test A is carried out.
solution.
Test solution. Shake a quantity of the powdered tablets Melt a sufficient quantity at 50°, prepare a thin film between
containing 0.1 g of Ethopropazine Hydrochloride with 50 ml of two previously warmed bromide plates and record the
chloroform for 15 minutes, centrifuge and use the supernatant spectrum immediately. Compare the spectrum with that
liquid. obtained with ethosuximide RS.
489
ETHOSUXIMIDE CAPSULES IP 2007
B. When examined in the range 230 nm to 360 nm (2.4.7), a inlet port and detector. 240°,
0.1 per cent w/v solution in ethanol (95 per cent) shows an – flow rate. 30 ml per minute of the carrier gas.
absorption maximum at about 248 nm; absorbance at 248 nm, Inject 1 µl of reference solution (c) and adjust the sensitivity
about 0.85. of the detector so that the heights of the three principal peaks
C. Dissolve 0.1 g in 3 ml of methanol and add 0.05 ml of a 10 are not less than 70 per cent of full-scale deflection. The peaks
per cent w/v solution of cobalt chloride, 0.05 ml of a 10 per in order of emergence, are due to 2-ethyl-2-methylsuccinic
cent w/v solution of calcium chloride and 0.1 ml of 2 M sodium acid, ethosuximide, and anthracene.
hydroxide; a purple colour develops and no precipitate is
The test is not valid unless the resolution factor between the
produced.
peaks corresponding to 2-ethyl-2-methylsuccinic acid and
Tests ethosuximide in the chromatogram obtained with test solution
(a) is at least 4.
Appearance of solution. Dissolve 2.5 g in sufficient water to
Inject 1 µl of test solution (a) and verify that there is no peak
produce 25 ml. The solution is clear (2.4.1) and colourless
with the same retention time as the internal standard. Inject
(2.4.1).
separately 1 µl of test solution (b) and reference solution (b)
Acidity. Dissolve 5.0 g in 50 ml of water by warming on a and record the chromatogram for twice the retention time of
water-bath for 5 minutes. Cool and titrate with 0.1 M sodium ethosuximide. Calculate the ratio (R) of the area of the peak
hydroxide using bromocresol green solution as indicator. due to ethosuximide to the area of the peak due to the internal
Not more than 0.7 ml of 0.1 M sodium hydroxide is required. standard in the chromatogram obtained with reference solution
Cyanide. Dissolve 1.0 g in 10 ml of ethanol (90 per cent) and (b). In the chromatogram obtained with test solution (b) the
add 0.5 ml of ferrous sulphate solution, 1 ml of 2 M sodium ratio of the sum of the areas of any secondary peaks to the
hydroxide and 0.1 ml of ferric chloride solution. Heat to area of the peak due to the internal standard is not greater
boiling, cool and acidify using 3 ml of 1 M sulphuric acid. than R.
After 15 minutes, there is no blue colour and no blue precipitate Sulphated ash (2.3.18). Not more than 0.1 per cent.
is produced.
Related substances. Determine by gas chromatography 1.0 g.
(2.4.13).
Assay. Weigh accurately about 0.12 g, dissolve 20 ml of
Test solution (a) Dissolve 1 g of the substance under dimethylformamide and add 0.2 ml of a 0.5 per cent w/v solution
examination in sufficient chloroform to produce 10 ml. of thymolphthalein in dimethylformamide. Titrate with 0.1 M
Test solution (b). Dilute 5 ml of test solution (a) to 10 ml with tetrabutylammonium hydroxide, until a distinct blue colour
a 0.01 per cent w/v solution of anthracene (internal standard) is produced.
in chloroform. 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
Reference solution (a). Dissolve 10 mg of 2-ethyl-2- 0.01412 g of C7H11NO2.
methylsuccinic acid in sufficient chloroform to produce Storage. Store protected from light.
10 ml.
Reference solution (b). Dilute 1 ml of test solution (a) to 100
ml with chloroform. To 1 ml of this solution add 5 ml of the Ethosuximide Capsules
internal standard solution and sufficient chloroform to produce
10 ml. Ethosuximide Capsules contain not less than 92.5 per cent
Reference solution (c). Dilute 1 ml of test solution (b) to 50 ml
ethosuximide, C7H11NO2.
with chloroform. Add 1 ml of this solution to 1 ml of reference
solution (a), add 5 ml of the internal standard solution and Identification
sufficient chloroform to produce 10 ml.
A. Heat a quantity of the contents of the capsules containing
0.1 g of Ethosuximide with 0.2 g of resorcinol and 0.1 ml of
– a glass column 2 m x 2 mm, packed with silanised
sulphuric acid at 140° for 5 minutes, add 5 ml of water, make
diatomaceous support (125 to 180 mesh) impregnated
alkaline with 5 M sodium hydroxide and add 0.2 ml to a large
with 3 per cent w/w polycyanopropylmethyl-
volume of water; a bright green fluorescence is produced.
phenylmethyl siloxane,
– temperature: B. Shake a quantity of the contents of the capsules containing
column.165°, 0.25 g of Ethosuximide with 80 ml of ethanol (95 per cent) for
490
IP 2007 ETHYLCELLULOSE
a few minutes, add sufficient ethanol (95 per cent) to produce each of 25 ml, of chloroform, washing each extract with the
100 ml, mix and filter. Dilute 20 ml of the filtrate to 100 ml with same 10 ml of water. To the combined extracts add 10 ml of a
ethanol (95 per cent). Absorbance of the resulting solution 3.0 per cent w/v solution of dimethyl phthalate (internal
at the maximum at about 248 nm, about 0.43 (2.4.7). standard) in chloroform, shake with 10 g of anhydrous sodium
sulphate and filter.
Tests
Other tests. Comply with the tests stated under Capsules. (a) but omit the internal standard.
Assay. Weigh accurately a quantity of the contents of the Reference solution. Add 2 ml of the internal standard solution
capsules containing about 0.2 g of Ethosuximide and dissolve to 25.0 ml of a 0.2 per cent w/v solution of ethosuximide RS in
in 30 ml of dimethylformamide. Titrate with 0.1 M chloroform.
tetrabutylammonium hydroxide, using a 0.1 per cent w/v Chromatographic system
solution of azo violet in dimethylformamide as indicator. Carry – a glass column 1.5 m x 4 mm, packed with acid-washed
out a blank titration. silanised diatomaceous support (80 to 100 mesh)
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to impregnated with 3 per cent w/w of cyanopropylmethyl
0.01412 g of C7H11NO2. phenyl methyl silicone fluid (OV-225) of
cyanopropylmethyl phenyl methyl silicone fluid (such
as OV-225),
exceeding 30°.
– temperature:
column.165°,
Ethosuximide Syrup – flow rate. 30 ml per minute of the carrier gas.
Determine the weight per ml of the syrup (2.4.29), and calculate
Ethosuximide Oral Solution
the content of C7H11NO2, weight in volume.
Ethosuximide Syrup is a solution of Ethosuximide in a suitable
Storage. Store at a temperature not exceeding 30°.
flavoured vehicle.
Ethosuximide Syrup contains not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of Ethylcellulose
ethosuximide, C7H11NO2.
Cellulose ethyl ether
Identification Ethylcellulose is an ethyl ether of cellulose.
A. Extract a quantity of the syrup containing 0.5 g of Ethylcellulose contains not less than 44.0 per cent and not
Ethosuximide with two quantities, each of 30 ml, of chloroform, more than 51.0 per cent of ethoxy (-OC2H5) groups, calculated
filter the combined extracts through a plug of cotton and on the dried basis.
evaporate the filtrate to dryness. Heat 100 mg of the residue
with 0.2 g of resorcinol and 0.1 ml of sulphuric acid at 140° for Description. A white to light tan powder; almost odourless.
5 minutes, cool, add 5 ml of water, make alkaline with 5 M Identification
sodium hydroxide and add 0.2 ml to a large volume of water;
a bright green fluorescence is produced. Dissolve 15 mg of the dried sample in 10 ml of dried
dichloromethane. Grind 0.5 ml of this solution to dryness
with 0.3 g of potassium bromide. The residue complies with
obtained with test solution (b) has the same retention time as
the following test.
that of the peak due to ethosuximide in the chromatogram
obtained with reference solution (a). Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with ethylcellulose
Tests RS.
Other tests. Complies with the tests stated under Oral liquids. Tests
Assay. Determine by gas chromatography (2.4.13). pH (2.4.24). 5.5 to 8.0, determined in a solution prepared in the
Test solution (a). Add 10 ml of water and 2 g of sodium following manner. Stir 1.0 g with 50 ml of carbon dioxide-free
bicarbonate to a weighed quantity of the syrup containing water previously heated to 90°, then cool and dilute with
about 0.25 g of Ethosuximide and extract with five quantities, sufficient carbon dioxide-free water to produce 100 ml.
491
ETHYL CHLORIDE IP 2007
Apparent viscosity. 90.0 to 110.0 per cent of that stated on the Ethyl Chloride
label for viscosity types of 10 millipascal seconds or more;
80.0 to 120.0 per cent of that stated on the label for viscosity CH3CH2Cl
types of 6 to 10 millipascal seconds; 75.0 to 140.0 per cent of
that stated on the label for viscosity types of 6 millipascal C2H5Cl Mol. Wt. 64.5
seconds or less, determined by the following method. Weigh Ethyl Chloride is chloroethane.
accurately about 0.5 g, calculated on the dried basis and
Description. Gaseous at ambient temperatures and pressures
dissolve in 95.0 ± 0.05 g of a mixture of 80 parts of toluene and
but is generally compressed to a colourless, mobile, flammable
20 parts of ethanol by weight. For ethylcellulose containing and very volatile liquid; odour, pleasant and ethereal.
less than 46.5 per cent of ethoxy groups use a mixture of
60 parts of toluene and 40 parts of ethanol. Determine the Identification
viscosity at 25° by Method A (2.4.28).
A. Burns with a luminous flame with the production of
Arsenic (2.3.10). Mix 1.0 g with 5 ml of sulphuric acid AsT, hydrogen chloride.
add a few glass beads and digest in a fumehood, preferably
on a hot plate at a temperature not exceeding 120°, until charring B. Hydrolyse a few ml with 5 M sodium hydroxide; the resulting
begins. (Additional acid may be necessary to wet some samples solution gives the reactions of chlorides (2.3.1), and after the
completely but the total volume added should not exceed 10 addition of iodine solution and warming, crystals of iodoform
ml). Cautiously add, dropwise, hydrogen peroxide solution are produced.
(30 per cent) allowing the reaction to subside and again
heating between additions of drops. Add the first few drops
Tests
very slowly with sufficient mixing to prevent a rapid reaction. Acidity or alkalinity. Shake 10 ml with 10 ml of ice-cold water
Discontinue heating if foaming becomes excessive. When the and allow the ethyl chloride to evaporate at room temperature;
reaction has abated, heat cautiously, rotating the flask the residual liquid (liquid A) is neutral to litmus solution.
occasionally to prevent the sample from caking on glass
Ionisable chlorides. 5 ml of liquid A gives no turbidity with
exposed to the heating unit. Maintain oxidising conditions
silver nitrate solution.
at all times during the digestion by adding small quantities
of the hydrogen peroxide solution whenever the mixture turns Ethanol. Warm 5 ml of liquid A with iodine solution and sodium
brown or darkens. Continue the digestion until the organic carbonate; no iodoform is produced.
matter is destroyed, gradually raising the temperature of the Distillation range. Into a dry 100-ml measuring cylinder insert
heating unit until fumes of sulphur trioxide are copiously a stopper carrying a short exit tube not less than 6 mm in
evolved and the solution becomes colourless or retains only internal diameter and an accurately standardised short-bulb
a light straw colour. Cool, add cautiously 10 ml of water, mix, thermometer covering the range –20° to +30° and graduated
and again evaporate till strong fuming, repeating this in tenths of a degree. Cover the bulb of the thermometer with
procedure to remove any trace of hydrogen peroxide. Cool, a piece of very fine muslin, free from grease and sizing materials,
add cautiously 10 ml of water, wash the sides of the flask with so that one end hangs down about 10 mm below the bulb.
a few ml of water, and dilute with water to 35 ml. The resulting Cool the cylinder in ice-water, transfer to it 100 ml of the sample,
solution complies with the limit test for arsenic (3 ppm). previously cooled in ice-water, insert the stopper and adjust
Heavy metals (2.3.13). 0.5 g complies with the limit test for the thermometer so that the end of the muslin dips into the
heavy metals, Method B (40 ppm). liquid and the bulb is above the surface. Replace the ice-water
with water at 24° to 26° and observe the temperature when 5
Sulphated ash (2.3.18). Not more than 0.5 per cent. ml of sample has evaporated and again when 5 ml remains.
Loss on drying (2.4.19). Not more than 3.0 per cent, determined Continually lower the thermometer to maintain its position
on 1.0 g by drying in an oven at 105° for 2 hours. relative to the liquid surface throughout the test. Correct the
observed temperature by adding 0.26° for every kPa that the
Assay. Weigh accurately about 50 mg in an empty, tared Hard barometric pressure is below 101.3 kPa or by subtracting 0.26°
Gelatin Capsule Shell and carry out the determination of for every kPa above. The corrected temperature is not lower
methoxyl (2.3.29). than 12.0° and not higher than 12.5°.
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.0007510 Other organic compounds. On evaporation, no foreign odour
of ethoxy (-OC2H5) groups. is detectable at any stage.
Labelling. The label states the apparent viscosity in mPa s of Non-volatile matter. Not more than 0.01 per cent w/w, when
a 2.0 per cent w/v solution and its ethoxy content. evaporated and dried at 105°.
492
IP 2007 ETHYLENEDIAMINE HYDRATE
Storage. Store protected from light in a refrigerator (2° to 8°). Ethylenediamine Hydrate
NH2 ,H2O
H2N
Ethyl Oleate C2H8N2,H2O Mol. Wt. 78.1
Ethylenediamine Hydrate is ethane-1,2-diamine
O
O CH3 monohydrate.
CH3 Ethylenediamine Hydrate contains not less than 97.5 per cent
w/w and not more than 101.5 per cent w/w of C2H8N2,H2O.
C20H38O2 Mol. Wt. 310.5 Description. A clear, colourless or slightly yellow liquid; odour,
ammoniacal.
Ethyl Oleate consists of the ethyl esters of (Z)-oleic and
related acids. Identification
Ethyl Oleate contains not less than 100.0 per cent w/w and not A. Dilute 1 ml to 6 ml with water. To 3 drops of the solution
more than 105.0 per cent w/w of the ethyl esters of (Z)-oleic add 2 ml of a 1 per cent w/v solution of copper sulphate and
and related acids, calculated as C20H38O2. shake; a purple-blue colour is produced.
Description. A pale yellow oil; odour, slight but not rancid. B. It is strongly alkaline.
Tests Tests
Weight per ml (2.4.29). 0.869 g to 0.874 g, determined at 20°. Ammonia and other bases. Weigh accurately about 1.5 ml and
Acid value (2.3.23). Not more than 0.5. transfer with the aid of ethanol (95 per cent) to a small dish.
Add, with stirring, 20 ml of dilute hydrochloric acid. Evaporate
Peroxides. Dissolve 5.0 g in 15 ml of chloroform, add 20 ml of the solution to dryness on a water-bath, breaking up any cake
glacial acetic acid and 0.5 ml of a saturated solution of formed with a glass rod, and dry at 105° for 1 hour.
potassium iodide, mix and allow to stand for exactly 1 minute
in the dark. Add 30 ml of water and titrate with 0.01 M sodium 1 g of residue is equivalent to 0.5872 g of C2H8N2,H2O.
thiosulphate using starch solution as indicator; not more Calculate the percentage of C2H8N2,H2O; the result is within
than 2.5 ml of 0.01 M sodium thiosulphate is required. 0.5 per cent of the percentage of ethylenediaminehydrate
Iodine value (2.3.28). 75 to 85. determined in the Assay.
Assay. Boil a suitable volume of ethanol (95 per cent) to Heavy metals (2.3.13). Evaporate 5.0 ml on a water-bath to
expel carbon dioxide and neutralise it to phenolphthalein dryness, add to the residue 1 ml of hydrochloric acid and
solution. Weigh accurately about 2.0 g of the substance under 0.5 ml of nitric acid and evaporate to dryness. Dissolve the
examination, dissolve in 5 ml of the neutralised ethanol residue in 20 ml of warm water, cool, add sufficient water to
contained in a hard-glass flask and neutralise the free acid in produce 100 ml and mix. 20 ml of the resulting solution complies
the solution with 0.1 M ethanolic potassium hydroxide using with the limit test for heavy metals, Method A (20 ppm).
0.2 ml of phenolphthalein solution as indicator. Add 25.0 ml Iron (2.3.14). To the residue obtained in the test for Non-
of 0.5 M ethanolic potassium hydroxide and boil under a volatile matter add 1 ml of hydrochloric acid and 0.5 ml of
reflux condenser on a water-bath for 1 hour and continue nitric acid and evaporate to dryness on a water-bath. Dissolve
boiling for 2 hours over a flame. Add 20 ml of water and titrate the residue in 20 ml of warm water and dilute with water to 100
the excess of alkali with 0.5 M hydrochloric acid using a ml. 40 ml of the solution complies with the limit test for iron
further 0.2 ml of phenolphthalein solution as indicator. Repeat (20 ppm).
the operation without the substance under examination. The
Non-volatile matter. Not more than 0.02 per cent w/v,
difference between the titres represents the alkali required to
determined on 5.0 ml by evaporating to dryness on a water-
saponify the substance under examination.
bath and drying at 105° for 1 hour.
1 ml of 0.5 M ethanolic potassium hydroxide is equivalent to
0.1553 g of C20H38O2.
water and titrate with 1 M hydrochloric acid using
Storage. Store protected from light in small, well-filled and bromophenol blue solution as indicator until a yellow colour
well-closed containers under an atmosphere of nitrogen. is produced.
493
ETHYLOESTRENOL IP 2007
1 ml of 1 M hydrochloric acid is equivalent to 0.03906 g of Tests

C2H8N2,H2O.
Storage. Store protected from light. in a 1.0 per cent w/v solution in dioxan.
17αα -Ethyloestran-17β β -ol. Determine by thin-layer
chromatography (2.4.17), coating the plate with silica gel G
Ethyloestrenol containing 20 per cent w/v of silver nitrate.
H3 C OH volumes of nonan-5-one.
CH3 Solvent mixture. A mixture of 9 volumes of chloroform and 1
H H
volume of methanol.
H H
C20H32O Mol. Wt. 288.5
Reference solution. A 0.08 per cent w/v solution of 17α−
Ethyloestrenol is 17α-ethylestr-4-en-17β-ol containing a ethyloestran-17β-ol RS in the same solvent mixture.
variable amount of methanol of crystallisation.
Ethyloestrenol contains not less than 95.0 per cent and not dry the plate, heat it at 105° for 10 minutes, spray with ethanolic
more than 103.0 per cent of C20H32O, calculated on the sulphuric acid (20 per cent), heat at 105° for a further 10
anhydrous and methanol-free basis. minutes and allow to cool. Any spot corresponding to 17α−
Description. A white or almost white, crystalline powder; ethyloestran-17β-ol in the chromatogram obtained with the
almost odourless. test solution is not more intense than the spot in the
Identification Related substances. Carry out Identification test B but using
A. Determine by infrared absorption spectrophotometry (2.4.6). 10 µl of the following solutions.
Compare the spectrum with that obtained with ethyloestrenol Test solution. A 1 per cent w/v solution of the substance
RS or with the reference spectrum of ethyloestrenol. under examination.
B. Determine by thin-layer chromatography (2.4.17), coating Reference solution (a). A 0.01 per cent w/v solution of the
the plate with silica gel G.. substance under examination.
Mobile phase. A mixture of 80 volumes of heptane and 20 Reference solution (b). A 0.005 per cent w/v solution of the
volumes of acetone. substance under examination.
Solvent mixture. A mixture of 90 volumes of chloroform and Any secondary spot in the chromatogram obtained with the
10 volumes of methanol. test solution is not more intense than the spot in the
Test solution. Dissolve 0.25 g of the substance under chromatogram obtained with reference solution (a) and not
examination in 100 ml with solvent mixture. more than one such spot is more intense than the spot in the
ethyloestrenol RS in the same solvent mixture. Methanol. Not more than 4.0 per cent w/w, determined by the
following method.
solution and reference solution (a). Determine by gas chromatography (2.4.13).
Apply to the plate 2 µl of each solution. After development, Test solution (a). A solution containing 10.0 per cent w/v of
dry the plate in air, heat it at 105° for 10 minutes, spray with the substance under examination in acetone.
ethanolic sulphuric acid (20 per cent) and heat at 105° for a
Test solution (b). A solution containing 10.0 per cent w/v of
further 10 minutes. Allow to cool and examine in daylight and
the substance under examination and 0.4 per cent v/v of
in ultraviolet light at 365 nm. The principal spot in the
ethanol (internal standard) in acetone.
that in the chromatogram obtained with reference solution (a). Reference solution. A solution containing 0.4 per cent v/v of
The principal spot in the chromatogram obtained with methanol and 0.4 per cent v/v of the internal standard in
reference solution (b) appears as a single, compact spot. acetone.
494
IP 2007 ETHYLOESTRENOL TABLETS
Chromatographic system Test solution. Extract a quantity of the powdered tablets

– a glass column 2.0 m x 0.4 mm, packed with porous containing 1 mg of Ethyloestrenol with chloroform, filter,
polymer beads (100 to 120 mesh) (such as Porapak Q), evaporate the filtrate to dryness at room temperature at a
– temperature: pressure not exceeding 0.2 kPa and dissolve the residue in
column.170°, 0.4 ml with solvent mixture.
inlet port and detector. 240°, Reference solution (a). A 0.25 per cent w/v solution of
– flame ionisation detector, ethyloestrenol RS in the same solvent mixture.
– flow rate. 30 ml per minute of the carrier gas (nitrogen).
Calculate the percentage w/w of methanol, assuming its weight solution and reference solution (a).
per ml at 20° to be 0.792 g.
Sulphated ash (2.3.18). Not more than 0.1 per cent. dry the plate in air, heat it at 105° for 10 minutes, spray with
Water (2.3.43). Not more than 0.5 per cent, determined on 5.0 ethanolic sulphuric acid (20 per cent) and heat at 105° for a
g. further 10 minutes. Allow to cool and examine in daylight and
in ultraviolet light at 365 nm. The principal spot in the
Assay. Determine by gas chromatography (2.4.13).
Test solution (a). A solution containing 0.2 per cent w/v of the that in the chromatogram obtained with reference solution (a).
substance under examination in chloroform. The principal spot in the chromatogram obtained with
Test solution (b). A solution containing 0.2 per cent w/v of the reference solution (b) appears as a single, compact spot.
substance under examination and 0.1 per cent w/v of arachidic B. In the Assay, the principal peak the chromatogram obtained
alcohol (internal standard) in chloroform. with the test solution has the same retention time as that of
Reference solution. A 0.2 per cent w/v solution of the peak due to ethyloestrenol RS in the chromatogram
ethyloestrenol RS in chloroform. obtained with the reference solution (a).
silanised diatomaceous support (80 to 100 mesh) coated 17αα -Ethyloestran-17β β -ol. Determine by thin-layer
with 3 per cent w/w of phenyl methyl silicone fluid chromatography (2.4.17), coating the plate with silica gel G
(50 per cent phenyl) (such as OV-17), containing 20 per cent w/v of silver nitrate.
– temperature: Mobile phase. A mixture of 75 volumes of toluene and 25
column.200°, volumes of nonan-5-one.
– flame ionisation detector, Solvent mixture. A mixture of 90 volumes of chloroform and
– flow rate. 30 ml per minute of the carrier gas (nitrogen). 10 volumes of methanol.
Calculate the content of C20H32O. Test solution. Dissolve 20 mg of the residue obtained in the
test for Related substances in 0.5 ml with solvent mixture.
Storage. Store protected from light in a refrigerator (2° to 8°).
Reference solution. A 0.08 per cent w/v solution of 17α-
ethyloestran-17β-ol RS in the same solvent mixture.
Ethyloestrenol Tablets dry the plate, heat it at 105° for 10 minutes, spray with ethanolic
Ethyloestrenol Tablets contain not less than 90.0 per cent and sulphuric acid (20 per cent), heat at 105° for a further 10
not more than 110.0 per cent of the stated amount of minutes and allow to cool. Any spot corresponding to 17α-
ethyloestrenol, C20H32O. ethyloestran-17β-ol in the chromatogram obtained with the
Identification chromatogram obtained with the reference solution.
A. Determine by thin-layer chromatography (2.4.17), coating Related substances. Carry out Identification test A but using
the plate with silica gel G. 10 µl of the following solutions.
Mobile phase. A mixture of 80 volumes of heptane and 20 Solvent mixture. A mixture of 90 volumes of chloroform and
volumes of acetone. 10 volumes of methanol.
Solvent mixture. A mixture of 90 volumes of chloroform and Test solution. Extract a quantity of the powdered tablets
10 volumes of methanol. containing 40 mg of Ethyloestrenol with chloroform, filter,
495
ETOPOSIDE IP 2007
evaporate the filtrate to dryness and dissolve 10 mg of the inlet port and detector. 280°,
residue in 1 ml with solvent mixture. – flame ionisation detector,
Reference solution (a). Dilute 1 volume of the test solution to – flow rate. 30 ml per minute of the carrier gas (nitrogen).
100 volumes with the same solvent mixture. Calculate the content of C20H32O in the tablets.
Reference solution (b). Dilute 1 volume of reference solution Storage. Store protected from light in a refrigerator (2° to 8°).
(a) to 2 volumes with the same solvent mixture.
Etoposide
H
H3 C O
Tablets. O O
Proceed as directed in the Assay using the following solutions. HO
OH O
Test solution. Powder one tablet, extract with 5 ml of chloroform O
in a centrifuge tube, centrifuge, evaporate 2.0 ml of the O
supernatant liquid in a current of nitrogen, dissolve the residue O
in 2 ml of acetone, evaporate the solution to dryness on a
O
water-bath and dissolve the residue in 0.4 ml of chloroform.
w/v of ethyloestranol RS and 0.1 per cent w/v of arachidic H3CO OCH3
alcohol (internal standard) in chloroform. OH
solution on 2.0 ml of the supernatant liquid but by extracting C29H32O13 Mol. Wt. 588.6
with 2.0 ml of a 0.02 per cent w/v solution of arachidic alcohol
Etoposide is 4′-demethylepipodophyllotoxin-9-[4,6-O-
in acetone.
ethylidene-β-D-glucopyranoside].
Calculate the content of C20H32O in the tablet. Etoposide contains not less than 95.0 per cent and not more
Other tests. Comply with the tests stated under Tablets. than 105.0 per cent of C29H32O13, calculated on the anhydrous
basis.
Test solution. Weigh and powder 20 tablets. Extract a quantity CAUTION — Etoposide is potentially cytotoxic. Great care
of the powdered tablets containing about 8 mg of should be taken in handling the powder and preparing
Ethyloestrenol with 20 ml of acetone, filter, evaporate the solutions.
filtrate to dryness on a water-bath and dissolve the residue in Description. A white or almost white crystalline powder.
4.0 ml of chloroform.
Identification
w/v of ethyloestrenol RS and 0.1 per cent w/v of arachidic Test A may be omitted if tests B and C are carried out. Tests B
alcohol (internal standard) in chloroform. and C may be omitted if test A is carried out.
Reference solution (b). Prepared in a similar manner as solution A. Determine by infrared absorption spectrophotometry (2.4.6).
(1) but extracting with 20 ml of a 0.02 per cent w/v solution of Compare the spectrum with that obtained with etoposide RS.
arachidic alcohol in acetone.
Chromatographic system the plate with silica gel H.
silanised diatomaceous support (80 to 100 mesh) coated Solvent mixture. 1 volume of methanol and 9 volumes of
with 3 per cent w/w of phenyl methyl silicone fluid dichloromethane.
(50 per cent phenyl) (such as OV-17), Mobile phase. A mixture of 1.5 volumes of water, 8 volumes of
– temperature: glacial acetic acid, 20 volumes of acetone and 100 volumes
column.200°, of dichloromethane.
496
IP 2007 ETOPOSIDE
Test solution. Dissolve 50 mg of the substance under Chromatographic system

examination in 10 ml of the solvent mixture. – a stainless steel column 25 cm x 4.6 mm, packed with
Reference solution. A 0.5 per cent w/v solution of etoposide octadecylsilane bonded to porous silica (5 µm),
RS in the solvent mixture. – column temperature 40º,
– mobile phase: A. a mixture of 1 volume of triethylamine,
Apply to the plate 5µl of each solution as bands 10 mm by 3 1 volume of anhydrous formic acid and 998 volumes of
mm. Allow the mobile phase to rise 17 cm. Dry the plate in water,
warm air, spray it with a mixture of 1 volume of sulphuric acid B. a mixture of 1 volume of triethylamine,
and 9 volumes of ethanol and heat at 140º for 15 minutes. The 1 volume of anhydrous formic acid and 998 volumes of
principal spot in the chromatogram obtained with the test acetonitrile,
solution corresponds to that in the chromatogram obtained – flow rate. 1 ml per minute,
with the reference solution. – a linear gradient programme using the conditions given
C. Dissolve 5 mg in 5 ml of glacial acetic acid and add 0.05 ml below,
of ferric chloride solution. Mix and cautiously add 2 ml of – spectrophotometer set at 285 nm,
sulphuric acid. Avoid mixing the 2 layers. Allow to stand for – a 10 µl loop injector.
about 30 minutes; a pink to reddish-brown ring develops at Time Mobile Mobile Comment
the interface and the upper layer is yellow. phase A phase B
Tests
0 75 25 isocratic
Appearance of solution. A 3.0 per cent w/v solution in a mixture 7 75 25 begin linear
of 1 volume of methanol and 9 volumes of dichloromethane gradient
is clear (2.4.1) and not more intensely coloured than reference 23 27 73 end chromatogram,
solution Y6 or BY6 (2.4.1). return to 75A
Specific optical rotation (2.4.22). – 106.0º to - 114.0º, determined 25 75 25 end equilibration,
in a 0.5 per cent w/v solution in a mixture of 1 volume of begin next
methanol and 9 volumes of dichloromethane. chromatogram
Related substances. Determine by liquid chromatography Inject reference solution (d). Continue the chromatography
(2.4.14). until the peak due to phenolphthalein is eluted. The
chromatogram shows two principal peaks corresponding to
Solvent mixture. Equal volumes of mobile phase A and mobile etoposide and to cis-etoposide. Ignore any peak due to
phase B. phenolphthalein.
Test solution (a). Dissolve 40 mg of the substance under Inject test solution (a) and reference solutions (a), (b) and (d).
examination in the solvent mixture and dilute to 10.0 ml with The resolution between the peaks due to etoposide and to
the solvent mixture. cis-etoposide is not less than 3.0. The area of any impurity
Test solution (b). Dissolve 50.0 mg of the substance under peak is not more than the area of the principal peak in the
examination in the solvent mixture and dilute to 50.0 ml with chromatogram obtained with reference solution (a) (0.5 per
the solvent mixture. cent) and not more than two such peaks have an area more
Reference solution (a). Dilute 1.0 ml of test solution (a) to 10.0
obtained with reference solution (b) (0.2 per cent), the sum of
ml with the solvent mixture. Dilute 1.0 ml of this solution to
the impurity peaks is not more than twice the area of the
20.0 ml with the solvent mixture.
Reference solution (b). Dilute 4.0 ml of reference solution (a) solution (a) (1 per cent). Ignore any peak that is less than 0.1
to 10.0 ml with the solvent mixture. times the area of the principal peak in the chromatogram
Reference solution (c) A 0.1 per cent w/v solution of etoposide obtained with reference solution (a).
RS in the solvent mixture. Heavy metals (2.3.13). 1.0 g complies with the limit test for
Reference solution (d) To 10 ml of test solution (b), add 0.1 ml heavy metals, Method B (20 ppm).
of a 4 per cent v/v solution of glacial acetic acid and 0.1 ml of Sulphated ash (2.3.18). Not more than 0.1 per cent.
phenolphthalein solution. Add 1 M sodium hydroxide until
the solution becomes faintly pink (about 0.15 ml). After 15 Water (2.3.43). Not more than 6.0 per cent, determined on
minutes, add 0.1 ml of a 4 per cent v/v solution of glacial 0.25 g.
acetic acid. Assay. Determine by liquid chromatography (2.4.14).
497
ETOPOSIDE CAPSULES IP 2007
Follow the chromatographic procedure described under Inject the reference solution. The test is not valid unless the
Related substances. resolution between the principal peak and the peak immediately
Inject reference solution (c). The relative standard deviation following the principal peak (cis-etoposide) is at least 1.0.
for replicate injections is not more than 2.0 per cent. Inject test solution (a). The area of any peak corresponding
to cis-etoposide is not more than the area of the peak in the
Inject alternately test solution (b) and reference solution (c).
chromatogram obtained with test solution (b) (2 per cent).
Calculate the content of C29H32O13.
Apparatus No. 2
Medium. 900 ml of a pH 4.5 buffer prepared by dissolving 2.99
g of sodium acetate and 14 ml of 2 M acetic acid in 1000 ml of
Etoposide Capsules water.
Etoposide Capsules contain not less than 90.0 per cent and Speed and time. 50 rpm and 45 minutes.
not more than 110.0 per cent of the stated amount of etoposide, Withdraw a suitable volume of the medium and filter. Determine
C29H32O13. by liquid chromatography (2.4.14).
CAUTION —Etoposide is potentially cytotoxic. Great care Test solution. Use the filtrate.
should be taken in handling the powder and preparing
solutions. Reference solution (a). A 0.005 per cent w/v solution of
etoposide RS in the dissolution medium.
Identification Reference solution (b). A solution containing 0.005 per cent
w/v of etoposide RS and 0.00025 per cent w/v of ethyl
Add a quantity of the contents of the capsules containing 0.1
parahydroxybenzoate in the dissolution medium.
g of Etoposide to a separating funnel containing 100 ml of
water,extract with two quantities, each of 20 ml, of Chromatographic system
dichloromethane, dry the combined organic extracts over – a stainless steel column 30 cm × 3.9 mm, packed with
anhydrous sodium sulphate and filter. Extract the filtrate with phenyl silica gel for chromatography (10 µm)(such as
30 ml of water, filter the dichloromethane layer through Bondapak phenyl),
anhydrous sodium sulphate and evaporate to dryness at 25º – mobile phase: a mixture of 26 volumes of acetonitrile
to 35º under reduced pressure. Dissolve the oily residue in 5 and 74 volumes of a 0.272 per cent w/v solution of sodium
ml of water, shake gently and allow to stand for 30 minutes. acetate adjusted to pH 4.0 with glacial acetic acid,
Filter through a sintered-glass funnel, wash the precipitate in – flow rate. 1 ml per minute,
the funnel with three quantities, each of 20 ml, of water and – spectrophotometer set at 254 nm,
dry the precipitate in the funnel at 40º at a pressure of 2 kPa for – a 20 µl loop injector.
90 minutes. The residue complies with the following test. D. Not less than 80 per cent of the stated amount of C29H32O13.
Compare the spectrum with that obtained with etoposide RS.
Tests Test solution. Weigh accurately a quantity of the mixed
cis-Etoposide. Determine by liquid chromatography (2.4.14). contents of 20 capsules containing about 40 mg of Etoposide
dissolve in the mobile phase and dilute to 100.0 ml with the
Test solution (a). Weigh accurately a quantity of the mixed mobile phase; use immediately.
contents of 20 capsules containing about 0.5 g of Etoposide,
dissolve in the mobile phase and dilute to 100 ml with the Reference solution (a). A 0.04 per cent w/v solution of
mobile phase; use immediately. etoposide RS in the mobile phase.
Test solution (b). Dilute 1 ml of test solution (a) to 50 ml with Reference solution (b). A solution containing 0.005 per cent
the mobile phase. w/v of etoposide RS and 0.00025 per cent w/v of ethyl
parahydroxybenzoate in the mobile phase.
Reference solution. A 0.5 per cent w/v solution of etoposide
RS in a mixture of 50 volumes of acetonitrile, 50 volumes of Use the chromatographic system described under Dissolution.
water and 0.1 volume of triethylamine and allow to stand for Inject reference solution (b). The test is not valid unless the
40 minutes. resolution between the two principal peaks is at least 2.0.
Use the chromatographic system described under Dissolution. Inject alternately the test solution and reference solution (a).
498
IP 2007 ETOPOSIDE INJECTION
Calculate the content of C29H32O13 in the capsules. B. In the Assay, the principal peak in the chromatogram
Storage. Store protected from moisture at a temperature not obtained with the test solution corresponds to the peak in the
exceeding 30º. The capsules should not be stored in a chromatogram obtained with the reference solution.
refrigerator.
Tests
pH (2.4.24). 3.0 to 4.0, determined in a solution prepared by
diluting a volume of the concentrate containing 0.1 g of
Etoposide Injection Etoposide to 50 ml with carbon dioxide-free water.
Etoposide Injection is a sterile material consisting of Etoposide cis-Etoposide. Determine by liquid chromatography (2.4.14).
Concentrate. It is filled in a sealed container.
Test solution. Dilute a volume of the concentrate containing
0.5 g of Etoposide to 100 ml with the mobile phase.
Injections in accordance with the manufacturer’s instructions, Reference solution (a). Dilute 1.0 ml of the test solution to 50
immediately before use. ml with the mobile phase.
The constituted solution complies with the requirements for Reference solution (b). A 0.5 per cent w/v solution of
Clarity of solution and Particulate matter stated under etoposide RS in the mobile phase.
Parenteral Preparations (Infusions). Use the chromatographic system described under Assay.
resolution between the principal peak and the peak immediately
after the principal peak (cis-etoposide) is not less than 1.0.
Etoposide Concentrate Inject the test solution and reference solution (a). The area of
any peak corresponding to cis-etoposide is not more than the
Etoposide Concentrate is a sterile solution of Etoposide in a
suitable ethanolic vehicle.
The concentrate complies with the requirements stated under
Parenteral Preparations (Concentrated Solutions for
Injection) and with the following requirements. Test solution. Dilute a volume of the concentrate containing
about 40 mg of Etoposide to 100.0 ml with the mobile phase.
Etoposide Concentrate contains not less than 95.0 per cent
and not more than 105.0 per cent of the stated amount of Reference solution. A 0.04 per cent w/v solution of etoposide
etoposide, C29H32O13. RS in the mobile phase.

Determine by thin-layer chromatography (2.4.17), coating the phenyl silica gel (10 µm)(such as ìBondapak phenyl),
plate with silica gel GF254. – mobile phase: a mixture of 26 volumes of acetonitrile
Mobile phase. A mixture of 80 volumes of dichloromethane, and 74 volumes of a 0.272 per cent w/v solution of sodium
25 volumes of acetone, 2.5 volumes of ethanol (95 per cent) acetate adjusted to pH 4.0 with glacial acetic acid,
and 0.5 volume of water. – flow rate. 1 ml per minute,
Test solution. Dilute a volume containing 20 mg of Etoposide – a 20 µl loop injector.
to 25 ml with a mixture of 9 volumes of dichloromethane and
1 volume of methanol. Inject the reference solution. The test is not valid unless the
Reference solution. A 0.08 per cent w/v solution of etoposide than 2.0 per cent.
RS in a mixture of 9 volumes of dichloromethane and 1 volume
of methanol. Inject alternately the test solution and the reference solution.
Apply to the plate 10 µl of each solution. Allow the mobile Calculate the content of C29H32O13 in the concentrate.
phase to rise 17 cm. Dry the plate in air, and examine in ultraviolet Storage. Store protected from light.
obtained with the test solution corresponds to that in the Labelling. The label states: (1) the directions for dilution of
chromatogram obtained with the reference solution. the contents; (2) that the diluted injection is to be given by
499
ETOPOSIDE INJECTION IP 2007
intravenous injection; (3) that the concentrate should be

500
F
Ferrous Fumarate ....
Ferrous Fumarate Tablets ....
Ferrous Gluconate ....
Ferrous Gluconate Tablets ....
Ferrous Sulphate ....
Dried Ferrous Sulphate ....
Ferrous Sulphate Tablets ....
Fludrocortisone Acetate ....
Fludrocortisone Tablets ....
Fluocinolone Acetonide ....
Fluocinolone Cream ....
Fluorescein Sodium ....
Fluorescein Eye Drops ....
Fluorouracil ....
Fluorouracil Injection ....
Fluphenazine Decanoate ....
Fluphenazine Decanoate Injection ....
Fluphenazine Hydrochloride ....
Fluphenazine Hydrochloride Injection ....
Fluphenazine Tablets ....
Fluticasone Propionate ....
Fluticasone Propionate Inhalation ....
Fluticasone Propionate powder for Inhalation ....
Flurbiprofen ....
Flurbiprofen Tablets ....
Folic Acid ....
Folic Acid Tablets ....
Formeterol Fumarate Dihydrate ....
Formeterol Fumarate and Budesonide powder for Inhalation ....
Framycetin Sulphate ....
501
Fructose ....
Fructose Injection ....
Frusemide ....
Frusemide Injection ....
Frusemide Tablets ....
Furazolidone ....
Furazolidone Oral Suspension ....
Furazolidone Tablets ....
Fusidic Acid ....
Fusidic Acid Oral Suspension ....
502
IP 2007 FERROUS FUMARATE TABLETS
citric acid, make alkaline with 5 M ammonia and add 1 ml of

Ferrous Fumarate potassium cyanide solution. Dilute to 50 ml with water and
O add 0.1 ml of sodium sulphide solution. Any brown colour
O produced is not more intense than that produced by treating
Fe O 1.0 ml of lead standard solution (20 ppm Pb) in a similar
O manner.
C4H2FeO4 Mol. Wt. 169.9 Sulphates (2.3.17). Boil 0.15 g with 10 ml of 2 M hydrochloric
acid and 20 ml of freshly boiled and cooled water, cool in ice
Ferrous Fumarate contains not less than 93.0 per cent and not
and filter; the filtrate complies with the limit test for sulphates
more than 101.0 per cent of C4H2FeO4, calculated on the dried
(0.1 per cent).
basis.
Ferric iron. Not more than 2.0 per cent, determined by the
Description. A reddish orange to reddish brown, fine powder;
following method. Weigh accurately about 3.0 g and dissolve
may contain soft lumps that produce a yellow streak when
in a mixture of 200 ml of water and 20 ml of hydrochloric acid
crushed; odour, slight.
by heating rapidly to boiling point. Boil for 15 seconds, cool
Identification rapidly, add 3 g of potassium iodide, close the flask, allow to
stand in the dark for 15 minutes and titrate the liberated iodine
A. Heat 1 g with 25 ml of a mixture of equal volumes of with 0.1 M sodium thiosulphate using starch solution, added
hydrochloric acid and water on a water-bath for 15 minutes, towards the end of the titration, as indicator. Repeat the
cool and filter. Reserve the precipitate for test B; the filtrate operation without the substance under examination. The
gives reaction A of ferrous salts (2.3.1). difference between the titrations represents the amount of
B. Wash the precipitate reserved in test A with a mixture of 1 iodine liberated by the ferric iron.
volume of dilute hydrochloric acid and 9 volumes of water 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.005585 g
and dry at 105°. Suspend 0.1 g of the residue in 2 ml of sodium of ferric iron.
carbonate solution and add dilute potassium permanganate
solution dropwise; the permanganate is decolorised and a Loss on drying (2.4.19). Not more than 1.0 per cent, determined
brownish solution is obtained. on 1.0 g by drying in an oven at 105°.
C. Mix 0.5 g with 1 g of resorcinol. To 0.5 g of the mixture in a Assay. Weigh accurately about 0.3 g and dissolve in 15 ml of
crucible, add a few drops of sulphuric acid and heat gently; a dilute sulphuric acid with the aid of gentle heat. Cool, add 50
deep red semi-solid mass is formed. Add the mass to a large ml of water and immediately titrate with 0.1 M ceric ammonium
volume of water; an orange-yellow solution without any sulphate using ferroin sulphate solution as indicator.
fluorescence is obtained. 1 ml of 0.1 M ceric ammonium sulphate is equivalent to 0.01699
g of C4H2FeO4.
Tests
Arsenic (2.3.10). Mix 2.0 g with 1.5 g of anhydrous sodium
carbonate, add 10 ml bromine water and mix thoroughly.
Evaporate to dryness on a water-bath, ignite gently and
Ferrous Fumarate Tablets
dissolve the cooled residue in 20 ml of brominated Ferrous Fumarate Tablets contain not less than 90.0 per cent
hydrochloric acid and 10 ml of water. Transfer to a small and not more than 105.0 per cent of the stated amount of
flask, add sufficient stannous chloride solution AsT to remove ferrous fumarate, C4H2FeO4. The tablets may be coated.
the yellow colour, connect to a condenser and distil 22 ml. The
distillate complies with the limit test for arsenic (5 ppm). Identification
Heavy metals. Not more than 20 ppm, determined by the The powdered tablets comply with the following tests.
following method. Ignite 1.0 g gently until free from carbon,
A. Heat 1 g with 25 ml of a mixture of equal volumes of
dissolve in 5 ml of hydrochloric acid by heating on a water-
hydrochloric acid and water on a water-bath for 15 minutes,
bath and evaporate to dryness. Dissolve the residue in a
cool and filter. Reserve the precipitate for test B; the filtrate
mixture of 15 ml of hydrochloric acid, 4 ml of nitric acid and
gives reaction A of ferrous salts (2.3.1).
6 ml of water. Boil gently for 1 minute, cool and extract with
three quantities, each of 20 ml, of ether. If the aqueous layer is B. Wash the precipitate reserved in test A with a mixture of
more than slightly yellow, extract with a fourth quantity of 1 volume of dilute hydrochloric acid and 9 volumes of water
20 ml of ether and reject the ether extracts, heat the aqueous and dry at 105°. Suspend 0.1 g of the residue in 2 ml of sodium
solution gently to remove the dissolved ether, add 1 g of carbonate solution and add dilute potassium permanganate
503
FERROUS GLUCONATE IP 2007
solution dropwise; the permanganate is decolorised and a Ferrous Gluconate contains not less than 95.0 per cent and
brownish solution is obtained. not more than 102.0 per cent of C12H22FeO14, calculated on the
C. Mix 0.5 g with 1 g of resorcinol. To 0.5 g of the mixture in a dried basis.
crucible, add a few drops of sulphuric acid and heat gently; a Description. A yellowish grey or pale greenish-yellow, fine
deep red semi-solid mass is formed. Add the mass to a large powder or granules; odour, slight, resembling that of burnt
volume of water; an orange-yellow solution without any sugar.
fluorescence is obtained.
Identification
Tests A. Dissolve 5 g in carbon dioxide-free water at 60°, cool and
Ferric iron. Weigh accurately a quantity of the powder dilute to 50 ml with water. 1 ml of the resulting solution gives
prepared for the Assay, containing about 1.5 g of Ferrous reaction A of ferrous salts (2.3.1).
Fumarate, in a stoppered flask, dissolve as completely as B. To 0.75 g in a test-tube add 7.5 ml of warm water, add 1 ml of
possible with the aid of heat in a mixture of 100 ml of freshly glacial acetic acid and 1 ml of freshly distilled
boiled and cooled water and 10 ml of hydrochloric acid, boil phenylhydrazine. Heat the mixture on a water-bath for
for 15 seconds, cool rapidly, add 3 g of potassium iodide, 30 minutes. Cool and scratch the inner surface of the test tube
close the flask and allow to stand in the dark for 15 minutes. with a glass rod until crystals of gluconic acid phenylhydrazide
Titrate the liberated iodine with 0.1 M sodium thiosulphate begin to form. Set aside for 10 minutes, filter, dissolve the
using starch solution, added towards the end of the titration, precipitate in hot water, mix a small amount of decolorising
as indicator. Repeat the operation without the substance under charcoal and filter into a test tube. Allow the filtrate to cool,
examination. The difference between the titrations represents and scratch the inner surface of the test tube; white crystals
the amount of iodine liberated by the ferric iron. Not more are obtained which melt at about 202°, with decomposition
than 13.5 ml of 0.1 M sodium thiosulphate is required. (2.4.21).
Disintegration (2.5.1). 60 minutes.
Tests
Appearance of solution. Dissolve 5.0 g in carbon dioxide-free
Assay. Weigh and powder 20 tablets. Weigh accurately a water at 60°, cool and dilute to 50 ml with the same solvent
quantity of the powder containing about 0.3 g of Ferrous (solution A). Dilute 2 ml of solution A to 10 ml with water.
Fumarate and dissolve in 15 ml of dilute sulphuric acid with When examined against the light, the resulting solution is
the aid of gentle heat. Cool, add 50 ml of water and immediately clear (2.4.1).
titrate with 0.1 M ceric ammonium sulphate using ferroin
sulphate solution as indicator. pH (2.4.24). 4.0 to 5.5, determined in solution A, 3 to 4 hours
after preparation.
1 ml of 0.1 M ceric ammonium sulphate is equivalent to 0.01699
g of C4H2FeO4. Arsenic (2.3.10). To 5.0 g add 15 ml of water and 15 ml of
stannated hydrochloric acid, distil 22 ml and add to the
Labelling. The label states the quantity of the active ingredient distillate 40 ml of water and 0.2 ml of stannous chloride solution
both as the amount of Ferrous Fumarate and in terms of the AsT. The resulting solution complies with the limit test for
equivalent amount of ferrous iron in each tablet. arsenic (2 ppm).
Heavy metals (2.3.13). Warm 2.0 g gently with 10 ml of nitric
Ferrous Gluconate acid until reaction begins and allow to stand until the evolution
of nitrous fumes subsides. Boil gently to complete oxidation,
adding a further 5 ml of nitric acid, if necessary, and continue
COO boiling until the volume is reduced to about 5 ml. Add 20 ml of
H OH hydrochloric acid, boil gently for 1 minute, cool and extract
with three quantities, each of 20 ml, of ether. If the acid solution
HO H
Fe is still more than faintly yellow, extract with a fourth quantity
H OH of 20 ml of ether and discard the ether extracts. Transfer the
H OH acid solution to a narrow-necked flask, rinse the separator
OH with 5 ml of water, and add the rinsings to the flask. Heat to
2 remove the dissolved ether and part of the hydrochloric acid.
Cool and dilute to 50 ml with water. 25 ml of the resulting
C12H22FeO14,xH2O Mol. Wt. 446.1 (anhydrous) solution complies with the limit test for heavy metals, Method
Ferrous Gluconate is ferrous di(D-gluconate). A (20 ppm).
504
IP 2007 FERROUS GLUCONATE TABLETS
Chlorides (2.3.12). 0.4 g complies with the limit test for chlorides 1 ml of 0.1 M ceric ammonium nitrate is equivalent to 0.04461
(625 ppm). g of C12H22FeO14.
Sulphates (2.3.17). 0.3 g complies with the limit test for Storage. Store protected from light.
Barium. Dissolve 0.1 g in 50 ml of distilled water, and 5 ml of
dilute sulphuric acid, and allow to stand for 5 minutes. The
solution is not more opalescent than a mixture of 10 ml of
Ferrous Gluconate Tablets
solution A and 45 ml of distilled water, when examined against Ferrous Gluconate Tablets contain not less than 90.0 per cent
the light. and not more than 105.0 per cent of the stated amount of
ferrous gluconate, C12H22FeO14,2H2O. The tablets may be
Ferric iron. Not more than 1.0 per cent, determined by the
coated.
following method. Weigh accurately about 5.0 g, transfer to a
glass-stoppered flask and dissolve in a mixture of 100 ml of Identification
freshly boiled and cooled water and 10 ml of hydrochloric
acid. Add 3 g of potassium iodide, shake well and allow to Dissolve a quantity of the powdered tablets containing about
stand in the dark for 5 minutes. Titrate the liberated iodine 1 g of Ferrous Gluconate in 10 ml of water and filter; the filtrate
with 0.1 M sodium thiosulphate using starch solution, added complies with the following tests.
towards the end of the titration, as indicator. Repeat the A. 1 ml of the filtrate gives reaction A of ferrous salts (2.3.1).
operation without the substance under examination. The
difference between the titrations represents the amount of B. To 7.5 ml of the filtrate add 1 ml of glacial acetic acid and
iodine liberated by the ferric iron. 1 ml of freshly distilled phenylhydrazine. Heat the mixture on
a water-bath for 30 minutes. Cool and scratch the inner surface
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.005585 g of the test tube with a glass rod until crystals of gluconic acid
of ferric iron. phenylhydrazide begin to form. Set aside for 10 minutes, filter,
Oxalic acid. Dissolve 1 g in 5 ml of water, add 2 ml of dissolve the precipitate in hot water, mix a small amount of
hydrochloric acid and transfer to a separator. Extract with decolorising charcoal and filter into a test tube. Allow the
two quantities, each of 20 ml, of ether. Evaporate the combined filtrate to cool, and scratch the inner surface of the test tube;
ether extracts to dryness on a water-bath and dissolve the white crystals are obtained which melt at about 202°, with
residue in 5 ml of water. Add 0.05 ml of acetic acid and 3 ml of decomposition (2.4.21).
calcium chloride solution; no turbidity is produced. C. Shake a quantity of the powdered tablets containing 0.5 g
Reducing sugars. Dissolve 0.5 g in 10 ml of water and make of Ferrous Gluconate with 10 ml of dilute hydrochloric acid,
alkaline with dilute ammonia solution. Pass hydrogen filter and add to the filtrate 1 ml of barium chloride solution;
sulphide into the solution and allow to stand for 30 minutes. an opalescence may be produced but no precipitate is formed.
Filter and wash the precipitate with two quantities, each of
5 ml, of water. Combine the filtrate and the washings and
Tests
acidify with dilute hydrochloric acid. Add 2 ml of dilute Ferric iron. Weigh accurately a quantity of the powder
hydrochloric acid in excess. Boil the solution until the vapours prepared for the Assay, containing about 5.0 g of Ferrous
no longer darken lead acetate paper and, if necessary, boil Gluconate, in a stoppered flask, dissolve as completely as
further to concentrate the solution to about 10 ml. Cool and possible without the aid of heat in a mixture of 100 ml of freshly
add 10 ml of sodium carbonate solution, set aside for 5 boiled and cooled water and 10 ml of hydrochloric acid, add
minutes, filter and dilute the filtrate to 100 ml with water. To 5 3 g of potassium iodide, close the flask and allow to stand in
ml of the filtrate add 2 ml of potassium cupri-tartrate solution the dark for 5 minutes. Titrate the liberated iodine with 0.1 M
and boil for 1 minute; no red precipitate is formed. sodium thiosulphate using starch solution, added towards
Loss on drying (2.4.19). 5.0 per cent to 10.0 per cent, determined the end of the titration, as indicator. Repeat the operation
on 1.0 g by drying in an oven at 105°. without the substance under examination. The difference
between the titrations represents the amount of iodine liberated
Assay. Dissolve 0.5 g of sodium bicarbonate in a mixture of 70 by the ferric iron. Not more than 11.2 ml of 0.1 M sodium
ml of water and 30 ml of 1 M sulphuric acid. When thiosulphate is required.
effervescence ceases, add about 1.0 g, accurately weighed, of
the substance under examination, shake gently to dissolve Other tests. Comply with the tests stated under Tablets.
and titrate with 0.1 M ceric ammonium nitrate, using 0.1 ml of Assay. Weigh and powder 20 tablets. Weigh accurately a
ferroin solution as indicator, until the red colour disappears. quantity of the powder containing about 1.0 g of Ferrous
505
FERROUS SULPHATE IP 2007
Gluconate, dissolve in a mixture of 140 ml of water and 60 ml of tetrachloride extracts and add sufficient carbon tetrachloride
1 M sulphuric acid and titrate with 0.1 M ceric ammonium to produce 100.0 ml. The resulting solution is not more intensely
sulphate, using ferroin solution as indicator, until the red coloured than a solution prepared by treating 4.0 ml of copper
colour disappears. standard solution (10 ppm Cu) and 7.5 ml of water in the
same manner (50 ppm).
1 ml of 0.1 M ceric ammonium sulphate, is equivalent to 0.04822
g of C12H22FeO14,2H2O. Lead. Make 25.0 ml of solution B alkaline with dilute ammonia
solution, add 1 ml of potassium cyanide solution and sufficient
water to produce 50.0 ml. Add 0.1 ml of sodium sulphide
Labelling. The label states the quantity of the active ingredient solution; the solution is not more intensely coloured than a
both as the amount of Ferrous Gluconate and in terms of the mixture of 10 ml of hydrochloric acid, 0.5 ml of nitric acid,
equivalent amount of ferrous iron in each tablet. 5.0 ml of lead standard solution (20 ppm Pb), 0.1 ml of sodium
sulphide solution and sufficient water to produce 50.0 ml
(50 ppm).
Zinc. To 2.5 ml of solution B add 1 g of citric acid and 1 g of
Ferrous Sulphate resorcinol, neutralise the solution with dilute ammonia
solution using thymol blue solution as indicator and shake
FeSO4,7H2O Mol. Wt. 278.0 for 1 minute with two quantities, each of 20 ml, of dithizone
Ferrous Sulphate contains not less than 98.0 per cent and solution. To the combined extracts add 10 ml of 0.1 M
not more than 105.0 per cent of FeSO4,7H2O. hydrochloric acid and shake for 1 minute. Separate the acid
layer, add 3 ml of 1 M hydrochloric acid and 20 ml of ammonium
Description. Bluish green crystals or a light green, crystalline
chloride solution and adjust the volume to 50.0 ml with water.
powder; odourless. Efflorescent in air. On exposure to moist
Add 1.0 ml of potassium ferrocyanide solution and allow to
air, the crystals rapidly oxidise and become brown.
stand for 15 minutes. Any turbidity produced is not greater
than that developed in 15 minutes by the addition of 1.0 ml of
Identification potassium ferrocyanide solution to a freshly prepared mixture
Gives reaction A of ferrous salts and the reactions of sulphates of 10.0 ml of zinc standard solution (10 ppm Zn), 4 ml of 1 M
(2.3.1). hydrochloric acid, 20 ml of ammonium chloride solution and
sufficient water to produce 50.0 ml (500 ppm).
Tests Manganese. Dissolve 1.0 g in 40 ml of water, add 10 ml of
Appearance of solution. Dissolve 2.5 g in carbon dioxide-free nitric acid and boil until red fumes are evolved. Add 0.5 g of
water, add 0.5 ml of 1 M sulphuric acid and dilute to 50.0 ml ammonium persulphate and boil for 10 minutes. Discharge
with water (solution A). The solution is not more opalescent any pink colour by the dropwise addition of a 5 per cent w/v
than opalescence standard OS2 (2.4.1). solution of sodium sulphite and boil until any odour of sulphur
dioxide is eliminated. Add 10 ml of water, 5 ml of phosphoric
pH (2.4.24). 3.0 to 4.0, determined in solution A. acid and 0.5 g of sodium periodate, boil for 1 minute and
Arsenic (2.3.10). Dissolve 5.0 g in 10 ml of water, add 15 ml of allow to cool. The resulting solution is not more intensely
stannated hydrochloric acid and distil 20 ml. To the distillate coloured than that of a solution prepared at the same time and
add a few drops of bromine solution, remove the excess of in the same manner using 1.0 ml of 0.02 M potassium
bromine with a few drops of stannous chloride solution AsT permanganate in place of the substance under examination
and add 40 ml of water. The resulting solution complies with (0.1 per cent).
the limit test for arsenic (2 ppm). Chlorides (2.3.12). 20 ml of solution A complies with the limit
Copper. Dissolve 8.0 g in 40 ml of hydrochloric acid, add 10 test for chlorides (250 ppm).
ml of nitric acid and 15 ml of water, boil gently for 5 minutes
Assay. Dissolve 2.5 g of sodium bicarbonate in a mixture of
and cool. Shake with four quantities, each of 30 ml, of ether
150 ml of water and 10 ml of sulphuric acid. When
and discard the ether extracts. Heat the acid solution on a
effervescence ceases, add about 0.5 g of the substance under
water-bath to remove the dissolved ether, cool and add
examination, accurately weighed, shake gently to dissolve
sufficient water to produce 100.0 ml (solution B). To 10.0 ml of
and titrate with 0.1 M ceric ammonium nitrate, using 0.1 ml of
solution B add 1 g of citric acid, make alkaline with dilute
ferroin solution as indicator, until the red colour disappears.
ammonia solution, add 25 ml of water and 5 ml of sodium
diethyldithiocarbamate solution. Extract successively with 1 ml of 0.1 M ceric ammonium nitrate is equivalent to
5, 3 and 2-ml quantities of carbon tetrachloride, mix the carbon 0.02780 g of FeSO4,7H2O.
506
IP 2007 FERROUS SULPHATE TABLETS
Dried Ferrous Sulphate hydrochloric acid, 20 ml of ammonium chloride solution and

sufficient water to produce 50.0 ml (500 ppm).
Dried Ferrous Sulphate is Ferrous Sulphate from which a part
of the water of crystallisation has been removed by drying at Manganese. Dissolve 1.0 g in 40 ml of water, add 10 ml of
a temperature of 40°. nitric acid and boil until red fumes are evolved. Add 0.5 g of
ammonium persulphate and boil for 10 minutes. Discharge
Dried Ferrous Sulphate contains not less than 86.0 per cent any pink colour by the dropwise addition of a 5 per cent w/v
and not more than 90.0 per cent of FeSO4. solution of sodium sulphite and boil until any odour of sulphur
Description. A greyish white to buff coloured powder. dioxide is eliminated. Add 10 ml of water, 5 ml of phosphoric
acid and 0.5 g of sodium periodate, boil for 1 minute and
Identification allow to cool. The resulting solution is not more intensely
coloured than that of a solution prepared at the same time and
Gives reaction A of ferrous salts and the reactions of sulphates in the same manner using 1.0 ml of 0.02 M potassium
(2.3.1). permanganate in place of the substance under examination
(0.1 per cent).
Tests
Arsenic (2.3.10). Dissolve 3.3 g in 10 ml of water, add 15 ml of
Copper. Dissolve 8.0 g in 40 ml of hydrochloric acid, add 10 stannated hydrochloric acid and distil 20 ml. To the distillate
ml of nitric acid and 15 ml of water, boil gently for 5 minutes add a few drops of bromine solution, remove the excess of
and cool. Shake with four quantities, each of 30 ml, of ether bromine with a few drops of stannous chloride solution AsT
and discard the ether extracts. Heat the acid solution on a and add 40 ml of water. The resulting solution complies with
water-bath to remove the dissolved ether, cool and add the limit test for arsenic (3 ppm).
sufficient water to produce 100.0 ml (solution A). To 10.0 ml of Basic sulphate. 2.0 g dissolves slowly in a mixture of 7.5 ml of
solution A add 1 g of citric acid, make alkaline with dilute freshly boiled and cooled water and 0.5 ml of 0.5 M sulphuric
ammonia solution, add 25 ml of water and 5 ml of sodium acid, producing a solution that is not more than faintly turbid.
diethyldithiocarbamate solution. Extract successively with
5, 3 and 2 ml quantities of carbon tetrachloride, mix the carbon Assay. Weigh accurately about 0.5 g, dissolve in a mixture of
tetrachloride extracts and add sufficient carbon tetrachloride 30 ml of water and 20 ml of 1 M sulphuric acid and titrate with
to produce 100.0 ml. The resulting solution is not more intensely 0.1 M ceric ammonium sulphate using ferroin solution as
coloured than a solution prepared by treating 4.0 ml of copper indicator.
standard solution (10 ppm Cu) and 7.5 ml of water in the 1 ml of 0.1 M ceric ammonium sulphate is equivalent to 0.01519
same manner (50 ppm). g of FeSO4.
Lead. Make 25.0 ml of solution A alkaline with dilute ammonia
solution, add 1 ml of potassium cyanide solution and sufficient
water to produce 50.0 ml. Add 0.1 ml of sodium sulphide
solution; the solution is not more intensely coloured than a Ferrous Sulphate Tablets
mixture of 10 ml of hydrochloric acid, 0.5 ml of nitric acid, 5.0 Ferrous Sulphate Tablets contain Dried Ferrous Sulphate.
ml of lead standard solution (20 ppm Pb), 0.1 ml of sodium
sulphide solution and sufficient water to produce 50.0 ml Ferrous Sulphate Tablets contain not less than 80.0 per cent
(50 ppm). and not more than 90.0 per cent of the stated amount of dried
ferrous sulphate, FeSO4. The tablets are coated.
Zinc. To 2.5 ml of solution A add 1 g of citric acid and 1 g of
resorcinol, neutralise the solution with dilute ammonia Identification
solution using thymol blue solution as indicator and shake
for 1 minute with two quantities, each of 20 ml, of dithizone A. The powdered tablets give reaction A of ferrous salts (2.3.1).
solution. To the combined extracts add 10 ml of 0.1 M B. Extract the powdered tablets with 2 M hydrochloric acid
hydrochloric acid and shake for 1 minute. Separate the acid and filter. The filtrate gives reaction A of sulphates (2.3.1).
layer, add 3 ml of 1 M hydrochloric acid and 20 ml of ammonium
chloride solution and adjust the volume to 50.0 ml with water. Tests
Add 1.0 ml of potassium ferrocyanide solution and allow to
stand for 15 minutes. Any turbidity produced is not greater
than that developed in 15 minutes by the addition of 1.0 ml of Assay. Weigh and powder 20 tablets. Weigh accurately a
potassium ferrocyanide solution to a freshly prepared mixture quantity of the powder containing about 0.5 g of Dried Ferrous
of 10.0 ml of zinc standard solution (10 ppm Zn), 4 ml of 1 M Sulphate, dissolve in a mixture of 30 ml of water and 20 ml of
507
FLUDROCORTISONE ACETATE IP 2007
1 M sulphuric acid and titrate with 0.1 M ceric ammonium Reference solution (a). Dissolve 25 mg of fludrocortisone RS
sulphate using ferroin solution as indicator. in 10 ml of the solvent mixture.
1 ml of 0.1 M ceric ammonium sulphate is equivalent to 0.01519 Reference solution (b). Mix equal volumes of the test solution
g of FeSO4. and reference solution (a).
Labelling. The label states the quantity of the active ingredient Place the dry plate in a tank containing a shallow layer of the
both as the amount of Dried Ferrous Sulphate and in terms of solvent mixture, allow the solvent mixture to ascend to the
the equivalent amount of ferrous iron in each tablet. top, remove the plate from the tank and allow the solvent to
phase in the direction in which the aforementioned treatment
was done.
Fludrocortisone Acetate
O the solvent to evaporate, heat at 120° for 15 minutes and spray
O the hot plate with ethanolic sulphuric acid (20 per cent v/v).
H3 C O CH3
OH Heat at 120° for a further 10 minutes, allow to cool and examine
HO
H3C H spot in the chromatogram obtained with the test solution
F H
O
compact spot.
C23H31FO6 Mol. Wt. 422.5
Fludrocortisone Acetate is 9α-fluoro-11β,17α,21-trihydroxy- Tests
pregn-4-ene-3,20-dione 21-acetate.
Fludrocortisone Acetate contains not less than 96.0 per cent in a 1.0 per cent w/v solution in dioxan.
and not more than 104.0 per cent of C23H31FO6, calculated on
the dried basis. Light absorption (2.4.7). Absorbance of a 0.001 per cent w/v
solution in ethanol at the maximum at about 240 nm, 0.39 to
Description. A white or almost white, crystalline powder; 0.42.
odourless or almost odourless; hygroscopic.
Identification (2.4.14).
Test A may be omitted if tests B and C are carried out. Tests B Test solution. Dissolve 20 mg of the substance under
and C may be omitted if test A is carried out. examination in the mobile phase and dilute to 10 ml with the
mobile phase.
Compare the spectrum with that obtained with fludrocortisone Reference solution (a). Dissolve 2 mg of fludrocortisone
acetate RS or with the reference spectrum of fludrocortisone acetate RS and 2 mg of hydrocortisone acetate RS in the
acetate. mobile phase and dilute to 50 ml with the mobile phase.
B. To a warm 1 per cent w/v solution in methanol add an equal Reference solution (b). Dilute 1 ml of the test solution to 50 ml
volume of potassium cupri-tartrate solution; a red precipitate with the mobile phase.
is produced. Chromatographic system
C. Determine by thin-layer chromatography (2.4.17), coating – a stainless steel column 20 cm x 4.6 mm, packed with
the plate with silica gel G. octadecylsilyl silica gel (5 µm),
– mobile phase: a mixture of 35 volumes of
Solvent mixture. A mixture of 90 volumes of acetone and 10 tetrahydrofuran and 65 volumes of water,
volumes of formamide. – flow rate. 1 ml per minute,
Mobile phase. A mixture of 30 volumes of toluene and 10 – spectrophotometer set at 254 nm,
volumes of chloroform. – a 20 µl loop injector.
Test solution. Dissolve 25 mg of the substance under Equilibrate the column with the mobile phase for about 30
examination in 10 ml of the solvent mixture. minutes.
508
IP 2007 FLUDROCORTISONE TABLETS
Adjust the sensitivity of the system so that the height of the Mobile phase. A mixture of 30 volumes of toluene and 10
principal peak in the chromatogram obtained with reference volumes of chloroform.
solution (b) is 70 per cent to 90 per cent of the full scale of the Test solution. Shake a quantity of the powdered tablets
recorder. containing 1 mg of Fludrocortisone Acetate with 20 ml of
Inject reference solution (a). The retention times are: chloroform for 5 minutes, filter and evaporate the filtrate to
hydrocortisone acetate, about 8.5 minutes and fludrocortisone dryness. Dissolve the residue in 4 ml of a mixture of 90 volumes
acetate about 10 minutes. The test is not valid unless the of chloroform and 10 volumes of methanol.
resolution between the peaks corresponding to hydrocortisone Reference solution (a). Dissolve 25 mg of fludrocortisone RS
acetate and fludrocortisone acetate is at least 1.0. If this is not in 10 ml of the solvent mixture.
achieved, adjust the concentration of tetrahydrofuran in the
mobile phase. Increasing the concentration of tetrahydrofuran
reduces the retention times.
Place the dry plate in a tank containing a shallow layer of the
Inject the test solution and reference solution (b). Continue solvent mixture, allow the solvent mixture to ascend to the
the chromatography for twice the retention time of the principal top, remove the plate from the tank and allow the solvent to
peak. In the chromatogram obtained with the test solution: evaporate. Use within 2 hours, with the flow of the mobile
the area of any peak other than the principal peak is not greater phase in the direction in which the aforementioned treatment
than half the area of the principal peak in the chromatogram was done.
obtained with reference solution (b) (1.0 per cent) and the sum
of the areas of all such peaks is not greater than 0.75 times the Apply to the plate 20 µl of each solution. Allow the mobile
area of the principal peak in the chromatogram obtained with phase to rise 12 cm. Dry the plate in a current of warm air, allow
reference solution (b) (1.5 per cent). Ignore any peak due to the solvent to evaporate, heat at 120° for 15 minutes and spray
the solvent and any peak with an area less than 0.025 times the hot plate with ethanolic sulphuric acid (20 per cent v/v).
that of the principal peak in the chromatogram obtained with Heat at 120° for a further 10 minutes, allow to cool and examine
reference solution (b). in daylight and in ultraviolet light at 365 nm. The principal
Sulphated ash (2.3.18). Not more than 0.1 per cent. corresponds to that in the chromatogram obtained with
Loss on drying (2.4.19). Not more than 1.0 per cent, determined reference solution (a). The principal spot in the chromatogram
on 1.0 g by drying in an oven at 105°. obtained with reference solution (b) appears as a single,
compact spot.
Assay. Weigh accurately about 10 mg, dissolve in 50 ml of
ethanol and add sufficient ethanol to produce 100.0 ml. Dilute Tests
5.0 ml of this solution to 50.0 ml with ethanol and mix. Measure
the absorbance of the resulting solution (2.4.7) at the maximum
Tablets.
at about 238 nm. Calculate the content of C23H31FO6 taking 405
as the specific absorbance at 238 nm. Powder one tablet, disperse in 10 ml of water and extract with
three quantities, each of 5 ml, of chloroform. Filter the extracts
Storage. Store protected from light at a temperature not through a plug of cotton wool moistened with chloroform.
exceeding 30°. Evaporate the chloroform on a water-bath just to dryness.
Cool and dissolve the residue in 10.0 ml of ethanol. Measure
Fludrocortisone Tablets about 240 nm (2.4.7). Calculate the content of C23H31FO6 taking
Fludrocortisone Acetate Tablets Other tests. Comply with the tests stated under Tablets.
Fludrocortisone Tablets contain not less than 90.0 per cent Assay. Determine by liquid chromatography (2.4.14).
Test solution (a). Weigh and powder 20 tablets. Shake a
fludrocortisone acetate, C23H31FO6.
weighed quantity of the powdered tablets containing about
Identification 0.5 mg of Fludrocortisone Acetate with 2 ml of water for one
minute, add 8 ml of acetonitrile and shake on a mechanical
Determine by thin-layer chromatography (2.4.17), coating the shaker for 40 minutes. Dilute the mixture to 20.0 ml with
plate with silica gel G. acetonitrile, centrifuge and use the supernatant liquid.
Solvent mixture. A mixture of 90 volumes of acetone and 10 Test solution (b). Prepare in the same manner as test solution
volumes of formamide. (a) but using 4.0 ml of a solution containing 0.01 per cent w/v
509
FLUOCINOLONE ACETONIDE IP 2007
of norethisterone RS (internal standard) in acetonitrile and Mobile phase. A mixture of 30 volumes of toluene and 10
4.0 ml of acetonitrile in place of 8 ml of acetonitrile. volumes of chloroform.
Reference solution. Mix 20.0 ml of internal standard, 25.0 ml of Test solution. Dissolve 25 mg of the substance under
a 0.01 per cent w/v solution of fludrocortisone acetate RS in examination in 10 ml of the solvent mixture.
acetonitrile and 10 ml of water and dilute to 100.0 ml with Reference solution (a). Dissolve 25 mg of fludrocortisone RS
acetonitrile. in 10 ml of the solvent mixture.
Chromatographic system Reference solution (b). Mix equal volumes of the test solution
– a stainless steel column 20 cm x 4.6 mm, packed with and reference solution (a).
octadecylsilyl silica gel (such as Spherisorb ODS 1),
– mobile phase: a mixture of 40 volumes of acetonitrile Place the dry plate in a tank containing a shallow layer of the
and 60 volumes of water, solvent mixture, allow the solvent mixture to ascend to the
– flow rate. 2 ml per minute, top, remove the plate from the tank and allow the solvent to
– spectrophotometer set at 240 nm, evaporate. Use within 2 hours, with the flow of the mobile
– a 20 µl loop injector. phase in the direction in which the aforementioned treatment
was done.
Calculate the content of C23H31FO6 in the tablets.
Storage. Store protected from light. phase to rise 12 cm. Dry the plate in a current of warm air, allow
Fluocinolone Acetonide Heat at 120° for a further 10 minutes, allow to cool and examine
OH spot in the chromatogram obtained with the test solution
O CH3 corresponds to that in the chromatogram obtained with
H3 C O CH3 reference solution (a). The principal spot in the chromatogram
HO obtained with reference solution (b) appears as a single,
O
H3C H compact spot.
F H C. Carry out the procedure given in Test B but using solutions
prepared in the following manner.
O
F Test solution. Dissolve 10 mg in 1.5 ml of glacial acetic acid
in a separating funnel, add 0.5 ml of a 2 per cent w/v solution
C24H30F2O6 Mol. Wt. 452.5 of chromium trioxide and allow to stand for 30 minutes. Add
5 ml of water and 2 ml of dichloromethane and shake
Fluocinolone Acetonide is 6α,9α-difluoro-11β,21-dihydroxy- vigorously for 2 minutes. Allow to separate and use the lower
16α,17α -isopropylidenedioxypregna-1,4-diene-3,20-dione. layer.
Fluocinolone Acetonide contains not less than 96.0 per cent Reference solution (a). Prepare in the same manner as the test
and not more than 104.0 per cent of C24H30F2O6, calculated on solution but using 10 mg of fluocinolone acetonide RS.
the dried basis.
Description. A white or almost white, crystalline powder. solution corresponds to that in the chromatogram obtained
with reference solution (a).
Identification D. Heat 0.5 ml of chromic-sulphuric acid in a test-tube (5 cm
Test A may be omitted if tests B, C and D are carried out. Tests x about 6 mm) in a naked flame until white fumes are evolved;
C and D may be omitted if tests A and B are carried out. the solution wets the sides of the tube readily and there is no
greasiness. Add 2 or 3 mg of the substance under examination
A. Determine by infrared absorption spectrophotometry (2.4.6). and again heat in a naked flame until white fumes appear; the
Compare the spectrum with that obtained with fluocinolone solution does not wet the sides of the tube and does not pour
acetonide RS. easily from the tube.
the plate with silica gel G. Tests
Solvent mixture. A mixture of 90 volumes of acetone and 10 Specific optical rotation (2.4.22). +92.0° to +96.0°, determined
volumes of formamide. in a 1.0 per cent w/v solution in dioxan.
510
IP 2007 FLUOCINOLONE CREAM
Light absorption. Dissolve 15 mg in about 50 ml of ethanol ml of this solution to 100.0 ml with ethanol. Measure the
and dilute to 100.0 ml with ethanol. Dilute 10.0 ml of the absorbance of the resulting solution at the maximum at about
solution to 100.0 ml with ethanol. Absorbance of the resulting 238 nm. Calculate the content of C24H30F2O6 taking 355 as the
solution at the maximum at about 239 nm, 0.52 to 0.56 (2.4.7). specific absorbance at 238 nm.
Related substances. Determine by liquid chromatography Storage. Store protected from light.
(2.4.14).
examination in acetonitrile and dilute to 10 ml with the same
solvent.
Fluocinolone Cream
Reference solution (a). Dissolve 2.5 mg of fluocinolone Fluocinolone Acetonide Cream
acetonide RS and 2.5 mg of triamcinolone acetonide RS in 45 Fluocinolone Cream contains Fluocinolone Acetonide in a
ml of acetonitrile and dilute to 100 ml with water. suitable base.
Reference solution (b). Dilute 1 ml of the test solution to100 Fluocinolone Cream contains not less than 90.0 per cent and
ml with acetonitrile. not more than 110.0 per cent of the stated amount of
Chromatographic system fluocinolone acetonide, C24H30F2O6.
base-deactivated end-capped octadecylsilyl silica gel Identification
(5 µm),
– mobile phase: a mixture of 450 ml of acetonitrile and 500
ml of water, allowed to equilibrate, the volume adjusted
to 1000 ml with water and mixed again, Mobile phase. A mixture of 60 volumes of n-hexane, 40 volumes
– flow rate. 1 ml per minute, of chloroform, 10 volumes of methanol and
– spectrophotometer set at 238 nm, 1 volume of triethylamine.
– a 20 µl loop injector. Test solution. Disperse, by shaking a quantity of the cream
Adjust the sensitivity of the system so that the height of the containing 0.25 mg of Fluocinolone Acetonide in 2 ml of
principal peak in the chromatogram obtained with reference chloroform, add 10 ml of methanol, shake vigorously, cool in
solution (b) is not less than 50 per cent of the full scale of the ice for 15 minutes, centrifuge at 3000 rpm for 15 minutes, decant
recorder. the clear supernatant liquid, evaporate to dryness on a water-
bath in a current of nitrogen and dissolve the residue in 1 ml of
chloroform.
triamcinolone acetonide about 8.5 minutes and fluocinolone
acetonide about 10 minutes. The test is not valid unless the Reference solution. A 0.025 per cent w/v solution of
resolution between the peaks corresponding to triamcinolone fluocinolone acetonide RS in chloroform.
acetonide and fluocinolone acetonide is not less than 3.0.
Inject the test solution and reference solution (b). Continue dry the plate in air until the odour of the solvent is no longer
the chromatography for 4 times the retention time of detectable, heat at 105° for 5 minutes and spray whilst hot
fluocinolone acetonide. In the chromatogram obtained with with blue tetrazolium solution. The principal spot in the
the test solution the area of any peak other than the principal chromatogram obtained with the test solution corresponds to
peak is not greater than the area of the principal peak in the that in the chromatogram obtained with the reference solution.
cent) and the sum of the areas of all such peaks is not greater
obtained with the test solution has the same retention time as
than 2.5 times the area of the principal peak in the chromatogram
that of the peak due to Fluocinolone Acetonide in the
obtained with reference solution (b) (2.5 per cent). Ignore any
peak due to the solvent and any peak with an area less than
0.05 times that of the principal peak in the chromatogram
Tests
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Other tests. Complies with the tests stated under Creams.
on 1.0 g by drying in an oven at 105° for 3 hours. Assay. Determine by liquid chromatography (2.4.14).
Assay. Weigh accurately about 50 mg, dissolve in ethanol, For creams containing 0.025 per cent to 0.2 per cent w/w of
add sufficient ethanol to produce 50.0 ml and mix. Dilute 2.0 fluocinolone acetonide:
511
FLUORESCEIN SODIUM IP 2007
Test solution (a). Weigh accurately a quantity of the cream Reference solution. A solution containing 0.00625 per cent
containing about 2.5 mg of Fluocinolone Acetonide, add 60 ml w/v of fluocinolone acetonide RS and 0.00125 per cent w/v of
of a solution prepared by adding 80 ml of methanol to 20 ml of phenacetin in chloroform.
a 25 per cent w/v solution of lithium chloride and disperse by For creams containing 0.0025 per cent w/w of fluocinolone
shaking vigorously. Add 100 ml of cyclohexane, shake gently acetonide:
for 2 minutes and separate the lower, aqueous methanolic
layer, taking care to exclude any solid matter that separates at Test solution (a). Prepare as described above but using a
the interface. Repeat the extraction using a further 25 ml of the quantity of the cream containing about 0.25 mg of Fluocinolone
lithium chloride solution. To the combined extracts add a Acetonide.
solution containing 11 g of alum in 214 ml of water followed Test solution (b). Prepare in the same manner as test solution
by 50 ml of chloroform, shake vigorously for about 3 minutes, (a) but adding 1.0 ml of a 0.005 per cent w/v solution of
allow the layers to separate and filter the chloroform extract phenacetin (internal standard) to the chloroform solution
through filter paper ( such as Whatman No 1), previously before diluting to 10.0 ml.
moistened with chloroform, again excluding any solid matter Reference solution. A solution containing 0.0025 per cent w/v
at the interface. Repeat the extraction with 50- and 10-ml of fluocinolone acetonide RS and 0.0005 per cent w/v of
quantities of chloroform, filtering the extracts as before. phenacetin in chloroform.
Evaporate the combined extracts to dryness on a water-bath
in a current of nitrogen, dissolve the residue in 5 ml of
chloroform, transfer to a 10-ml volumetric flask with the aid of
porous silica particles (5 µm),
chloroform and add sufficient chloroform to produce 10.0 ml.
– mobile phase: a mixture of 58 volumes of n-hexane, 40
Test solution (b). Prepare in the same manner as test solution volumes of chloroform, 2 volumes of methanol and 0.1
(a) but adding 1.0 ml of a 0.05 per cent w/v solution of volume of glacial acetic acid,
phenacetin (internal standard) to the chloroform solution – flow rate. 1.8 ml per minute,
before dilution to 10.0 ml. – spectrophotometer set at 243 nm,
of fluocinolone acetonide RS and 0.005 per cent w/v of The assay is not valid unless the resolution between the peaks
phenacetin in chloroform. due to fluocinolone acetonide and phenacetin is more than 2,
and the capacity factors of fluocinolone acetonide and
For creams containing 0.01 per cent w/w of fluocinolone phenacetin are about 3 and 2, respectively. If these conditions
acetonide: are not achieved, adjust the concentration of methanol and
Test solution (a). Prepare as described above but using a chloroform in the mobile phase. Repeat the adjustment of
quantity of the cream containing about 1 mg of Fluocinolone chloroform and methanol concentration until correct values
Acetonide. for both resolution and capacity factors have been obtained.
Calculate the content of C24H30F2O6 in the cream.
(a) but adding 1.0 ml of a 0.02 per cent w/v solution of Storage. Store at a temperature not exceeding 30°.
phenacetin (internal standard) to the chloroform solution
before diluting to 10.0 ml.
Fluorescein Sodium
fluocinolone acetonide RS and 0.002 per cent w/v of Soluble Fluorescein
phenacetin in chloroform.
NaO O O
For creams containing 0.00625 per cent w/w of fluocinolone
acetonide:
Test solution (a). Prepare as described above but using a
quantity of the cream containing about 0.625 mg of COONa
Fluocinolone Acetonide.
(a) but adding 1.0 ml of a 0.0125 per cent w/v solution of C20H10Na2O5 Mol. Wt. 376.3
phenacetin (internal standard) to the chloroform solution Fluorescin Sodium is disodium 2-(3-oxo-6-oxido-3H-
before diluting to 10.0 ml. xanthen-9-yl)benzoate.
512
IP 2007 FLUORESCEIN SODIUM
Fluorescein Sodium contains not less than 98.5 per cent and Chlorides (2.3.12). Dissolve 75 mg in 20 ml of water, add 2 ml
not more than 100.5 per cent of C20H10Na2O5, calculated on the of nitric acid and filter; the filtrate complies with the limit test
dried basis. for chlorides (0.33 per cent).
Description. An orange-red powder; odourless or almost Sulphates (2.3.17). Dissolve 62.5 mg in 100 ml of water. To
odourless; hygroscopic. 20 ml add 2.5 ml of dilute hydrochloric acid and filter; the
filtrate complies with the limit test for sulphates (1.2 per cent).
Identification Dimethylformamide. Determine by gas chromatography
Compare the spectrum with that obtained with fluorescein Test solution (a). Dissolve 1.0 g of the substance under
sodium RS or with the reference spectrum of fluorescein examination in 10 ml of water, add, with stirring, 10 ml of 0.6 M
sodium. hydrochloric acid, allow to stand for 15 minutes and
B. A solution is strongly fluorescent, even in extreme dilutions. centrifuge. To 5 ml of the supernatant liquid add 0.1 g of
The fluorescence disappears when the solution is made acidic trisodium phosphate and shake to dissolve.
and reappears when it is made alkaline. Test solution (b). Prepare in the same manner as test solution
C. A drop of a 0.05 per cent w/v solution, absorbed on a piece (a) but using 10 ml of a 0.02 per cent w/v solution of
of filter paper, colours the paper yellow. On exposing the moist dimethylacetamide (internal standard) in place of water.
paper to the vapours of bromine for 1 minute and then to the Reference solution. Mix 10 ml of a 0.02 per cent w/v solution
vapours of ammonia, the yellow colour becomes deep pink. of dimethylformamide with 10 ml of the internal standard.
D. The residue after incineration gives the reactions of sodium Chromatographic system
salts (2.3.1). – a glass column 1.5m x 4 mm, packed with acid-washed,
Tests with 10 per cent w/w of polyethylene glycol 1000,
– temperature:
column.120°,
Related substances. Determine by thin-layer chromatography inlet port and detector. 180°,
(2.4.17), using a silica gel F254 precoated plate (such as Merck – flame ionisation detector,
silica gel 60 F254 plate). – flow rate. 30 ml per minute of the carrier gas (nitrogen).
Mobile phase. A mixture of 80 volumes of chloroform and 20 In the chromatogram obtained with test solution (b) the ratio
volumes of methanol. of the area of any peak corresponding to dimethylformamide
Test solution. Dissolve 0.1 g of the substance under to the area of the peak due to the internal standard is not
examination in 10 ml of 0.1 M methanolic hydrochloric acid. greater than the corresponding ratio in the chromatogram
substance under examination in 100 ml of 0.1 M methanolic Resorcinol. Determine by thin-layer chromatography (2.4.17),
hydrochloric acid. using a silica gel F254 precoated plate (such as Merck silica
gel 60 F254 plate).
dry the plate in air and examine in ultraviolet light at 365 nm. Mobile phase. A mixture of 60 volumes of hexane and 40
Expose the plate to iodine vapour for 30 minutes. Any volumes of ethyl acetate.
secondary spot in the chromatogram obtained with the test Test solution. Dissolve 1 g of the substance under examination
solution is not more intense than the spot in the chromatogram in 10 ml of water, add slowly with constant stirring, 10 ml of
obtained with the reference solution. 0.6 M hydrochloric acid, allow to stand for 15 minutes,
Chloroform-soluble matter. Dissolve 0.2 g in 10 ml of 0.1 M centrifuge and use the supernatant liquid.
sodium hydroxide and extract with 10 ml of chloroform. Dry Reference solution. A 0.025 per cent w/v solution of resorcinol
the chloroform layer with anhydrous sodium sulphate and in water.
filter. Absorbance of the resulting solution at about 480 nm,
using chloroform as the blank, not more than 0.10 (2.4.7).
dry the plate in air and expose to iodine vapour for 30 minutes.
Zinc. Dissolve 0.1 g in 10 ml of water, add 2 ml of hydrochloric Any spot corresponding to resorcinol in the chromatogram
acid, filter and add 0.1 ml of potassium ferrocyanide solution; obtained with the test solution is not more intense than the
no turbidity or precipitate is produced immediately. spot in the chromatogram obtained with the reference solution.
513
FLUORESCEIN EYE DROPS IP 2007
Acriflavin. Dissolve 10 mg in 5 ml of water, and add a few Related substances. Determine by thin-layer chromatography
drops of sodium salicylate solution; no precipitate is formed. (2.4.17), using a silica gel F254 precoated plate (such as Merck
Loss on drying (2.4.19). Not more than 10.0 per cent, silica gel 60 F254 plate).
determined on 1.0 g by drying in an oven at 105°. Mobile phase. A mixture of 80 volumes of chloroform and 20
volumes of methanol.
water, add 5 ml of dilute hydrochloric acid, and extract with Test solution. Dilute a suitable volume of the eye drops, if
four quantities, each of 20 ml, of a mixture of equal volumes of necessary, with an equal volume of 0.1 M methanolic
2-methyl-1-propanol and chloroform. Wash the combined hydrochloric acid so as it give a concentration of 1.0 per cent
extracts with 10 ml of water, extract the washings with 5 ml of w/v of fluorescein sodium.
the mixture of 2-methyl-1-propanol and chloroform and add Reference solution. Dilute 1 volume of the test solution to 500
to the combined extracts. Evaporate the combined extracts to volumes with 0.1 M methanolic hydrochloric acid.
dryness on a water-bath in a current of air, dissolve the residue
in 10 ml of ethanol (95 per cent), evaporate to dryness on a Apply to the plate 5 µl of each solution. After development,
water-bath and dry to constant weight at 105°. dry the plate in air and examine in ultraviolet light at 365 nm.
Expose the plate to iodine vapour for 30 minutes. Any
1 g of the residue is equivalent to 1.132 g of C20H10Na2O5. secondary spot in the chromatogram obtained with the test
Storage. Store protected from light. solution is not more intense than the spot in the chromatogram
Chloroform-soluble matter. To a volume of the eye drops
containing 0.1 g of Fluorescein Sodium add 1 ml of 2 M sodium
Fluorescein Eye Drops hydroxide, extract with 10 ml of chloroform, dry the chloroform
layer with anhydrous sodium sulphate and filter; absorbance
Fluorescein Sodium Eye Drops of the resulting solution at about 480 nm, using chloroform as
Fluorescein Eye Drops are a sterile solution of Fluorescein the blank, not more than 0.05 (2.4.7).
Sodium in Purified Water. Dimethylformamide. Determine by gas chromatography
Fluorescein Eye Drops contain not less than 90.0 per cent and (2.4.13).
not more than 110.0 per cent of the stated amount of Test solution (a). Dilute the eye drops with water, if necessary,
fluorescein sodium, C20H10Na2O5. to produce a solution containing 1.0 per cent w/v of
Fluorescein Sodium. To 5 ml of this solution add, with stirring,
Identification 0.3 ml of 1 M hydrochloric acid, allow to stand for 15 minutes
and centrifuge; dissolve 10 mg of trisodium phosphate in 2 ml
A. Evaporate a volume of the eye drops containing 20 mg of
of the supernatant liquid.
Fluorescein Sodium and dry at 105° for 30 minutes. The residue
complies with the following test. Test solution (b). Prepare in the same manner as test solution
(a) but adding 1.0 ml of a 0.01 per cent v/v solution of dimethyl-
Determine by infrared absorption spectrophotometry (2.4.6). acetamide (internal standard) before the hydrochloric acid.
Compare the spectrum with that obtained with fluorescein
sodium RS or with the reference spectrum of fluorescein Reference solution. A solution containing 0.002 per cent v/v
sodium. of dimethylformamide and 0.002 per cent v/v of the internal
standard.
B. Dilute the eye drops with water to produce a solution
containing 0.05 per cent w/v of Fluorescein Sodium. One drop Chromatographic system
of the solution, absorbed by a piece of filter paper, colours the – a glass column 1.5 m x 4 mm, packed with acid-washed,
paper yellow. On exposing the moist paper to the vapours of silanised diatomaceous support (80 to 100 mesh) coated
bromine for 1 minute and then to vapours of ammonia, the with 10 per cent w/w of polyethylene glycol 1000,
yellow colour becomes deep pink. – temperature:
column.120°,
C. The eye drops are strongly fluorescent, even in extreme inlet port and detector. 180°,
dilution. The fluorescence disappears when the solution is – flame ionisation detector,
made acidic and reappears when it is made alkaline. – flow rate. 30 ml per minute of the carrier gas (nitrogen).
Tests In the chromatogram obtained with test solution (b) the ratio
of the area of any peak corresponding to dimethylformamide
pH (2.4.24). 7.0 to 9.0. to the area of the peak due to the internal standard is not
514
IP 2007 FLUOROURACIL
greater than the corresponding ratio in the chromatogram Fluorouracil contains not less than 98.0 per cent and not more
obtained with the reference solution. than 101.0 per cent of C4H3FN2O2, calculated on the dried
Resorcinol. Determine by thin-layer chromatography (2.4.17), basis.
using a silica gel F254 precoated plate (such as Merck silica Description. A white or almost white, crystalline powder.
gel 60 F254 plate).
CAUTION - Great care should be taken to avoid inhaling
Mobile phase. A mixture of 60 volumes of hexane and 40 particles of Fluorouracil and exposing the skin to it.
volumes of ethyl acetate.
Identification
Test solution. Dilute the eye drops with water, if necessary, to
produce a solution containing 1.0 per cent w/v of Fluorescein Test A may be omitted if tests B, C and D are carried out. Tests
Sodium and to 10 ml add, with stirring, 2.5 ml of 0.25 M B, C and D may be omitted if test A is carried out.
hydrochloric acid. Allow to stand for 15 minutes, centrifuge
and dissolve 0.1g of trisodium phosphate in 5ml of the
Compare the spectrum with that obtained with fluorouracil
supernatant liquid.
RS or with the reference spectrum of fluorouracil.
Reference solution. A 0.004 per cent w/v solution of resorcinol B. When examined in the range 230 nm to 360 nm (2.4.7), a
in water. 0.001 per cent w/v solution in acetate buffer pH 4.7 shows an
Apply to the plate 5 µl of each solution. After development, absorption maximum only at about 266 nm.
dry the plate in air and expose to iodine vapour for 30 minutes. C. In the test for Related substances, the principal spot in the
Any spot corresponding to resorcinol in the chromatogram chromatogram obtained with test solution (b) corresponds to
obtained with the test solution is not more intense than the that in the chromatogram obtained with reference solution (a).
D. To 5 ml of a 1 per cent w/v solution add 1 ml of bromine
Other tests. Comply with the tests stated under Eye Drops. water; the colour of bromine is discharged.
Assay. Dilute an accurately measured volume of the eye drops
containing about 0.1 g of Fluorescein Sodium to 20 ml with
Tests
water. Add 5 ml of dilute hydrochloric acid and extract with Appearance of solution. A 1.0 per cent w/v solution in carbon
four quantities, each of 20 ml, of a mixture of equal volumes of dioxide-free water is clear (2.4.1), and not more intensely
2-methyl-1-propanol and chloroform. Wash the combined coloured than reference solution YS7 or BYS7 (2.4.1).
extracts with 10 ml of water, extract the washings with 5 ml of pH (2.4.24). 4.5 to 5.0, determined in a 1.0 per cent w/v solution.
the mixture of 2-methyl-1-propanol and chloroform and add to
the combined extracts. Evaporate the combined extracts to Related substances. Determine by thin-layer chromatography
dryness on a water-bath in a current of air, dissolve the residue (2.4.17), coating the plate with silica gel GF254.
in 10 ml of ethanol (95 per cent), evaporate to dryness on a Mobile phase. A mixture of 70 volumes of ethyl acetate,
water-bath and dry to constant weight at 105°. 15 volumes of methanol and 15 volumes of water.
1 g of the residue is equivalent to 1.132 g of C20H10Na2O5. Test solution (a). Dissolve 0.1 g of the substance under
examination in 10 ml of methanol (50 per cent).
Test solution (b). Dilute 5 ml of the test solution to 25 ml with
Labelling. The label states that the eye drops should be
methanol (50 per cent).
discarded after use on a single occasion.
fluorouracil RS in methanol (50 per cent).
Fluorouracil fluorouracil RS in methanol (50 per cent).
H Reference solution (c). A 0.0025 per cent w/v solution of
N O 5-hydroxyuracil in methanol (50 per cent).
NH Apply to the plate 10 µl of each solution. After development,

F dry the plate in air and examine in ultraviolet light at 254 nm.
O Any secondary spot in the chromatogram obtained with the
C4H3FN2O2 Mol. Wt. 130.1 chromatogram obtained with reference solution (b). Spray the
Fluorouracil is 5-fluoro-1H,3H-pyrimidine-2,4-dione. plate with a freshly prepared 0.5 per cent w/v solution of fast
515
FLUOROURACIL INJECTION IP 2007
blue B salt and then with 0.1 M sodium hydroxide. Any spot Related substances. Determine by thin-layer chromatography
corresponding to 5-hydroxyuracil in the chromatogram (2.4.17), coating the plate with silica gel GF254.
Mobile phase. A mixture of 70 volumes of ethyl acetate,15
(c). Ignore any secondary spot on or near the line of application.
Test solution. Dilute a suitable quantity of the injection with
water to produce a solution containing the equivalent of 2 per
cent w/v of Fluorouracil.
on 1.0 g in a platinum crucible. Reference solution (a). Dilute 1 volume of test solution to 400
volumes with methanol (50 per cent).
on 1.0 g by drying over phosphorus pentoxide in an oven at Reference solution (b). A 0.005 per cent w/v solution of
80° at a pressure of 1.5 to 2.5 kPa for 4 hours. 5-hydroxyuracil in methanol.
Assay. Weigh accurately about 0.2 g, dissolve in 80 ml of Apply to the plate 10 µl of each solution. After development,
dimethylformamide with the aid of gentle heat and cool. Titrate dry the plate in air and examine in ultraviolet light at 254 nm.
with 0.1 M tetrabutylammonium hydroxide in methanol, using Any secondary spot in the chromatogram obtained with the
0.25 ml of a 1 per cent w/v solution of thymol blue in test solution is not more intense than the spot in the
dimethylformamide as indicator. Carry out a blank titration. chromatogram obtained with reference solution (a). Spray with
a freshly prepared solution containing 0.5 per cent w/v of fast
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
blue B salt and then with 0.1 M sodium hydroxide. Any spot
0.01301 g of C4H3FN2O2.
corresponding to 5-hydroxyuracil in the chromatogram
Storage. Store protected from light. obtained with the test solution is not more intense than the
(b). Ignore any secondary spot on or near the line of
Fluorouracil Injection application.
Fluorouracil Injection is a sterile solution in Water for Injections Urea. Carry out the method described under Related
of fluorouracil sodium, prepared by the interaction of substances applying separately to the plate 20 µl of the
Fluorouracil and Sodium Hydroxide. following solutions. For the test solution dilute a suitable
quantity of the injection with water to produce a solution
Fluorouracil Injection contains not less than 90.0 per cent and containing the equivalent of 0.5 per cent w/v of Fluorouracil.
not more than 110.0 per cent of the stated amount of The reference solution contains 0.02 per cent w/v of urea in
fluorouracil, C4H3FN2O2. water. After development, dry the plate in air, spray with a
Description. A colourless or almost colourless solution. mixture of 10 volumes of a 1 per cent w/v solution of
4-dimethylaminobenzaldehyde in ethanol (95 per cent) and
Identification 1 volume of hydrochloric acid and heat at 105° until maximum
A. Acidify carefully a volume of the injection containing 0.1 g intensity of the spots is obtained. Any spot corresponding to
of Fluorouracil with glacial acetic acid, stir, cool and filter. urea in the chromatogram obtained with the test solution is
Wash the precipitate with 1 ml of water and dry over not more intense than the spot in the chromatogram obtained
phosphorus pentoxide at 80° at a pressure of 2 kPa for 4 hours. with the reference solution.
The residue complies with the following test. Other tests. Complies with the tests stated under Parenteral
Determine by infrared absorption spectrophotometry (2.4.6). Preparations (Injections).
Compare the spectrum with that obtained with fluorouracil Assay. To an accurately measured volume containing about
RS or with the reference spectrum of fluorouracil. 50 mg of Fluorouracil add 20 ml of 1 M hydrochloric acid and
B. When examined in the range 230 nm to 360 nm (2.4.7), the sufficient water to produce 250.0 ml. Dilute 5.0 ml to 100.0 ml
final solution obtained in the Assay shows an absorption with 0.1 M hydrochloric acid and measure the absorbance of
maximum only at about 266 nm. the resulting solution at the maximum at about 266 nm (2.4.7).
Calculate the content of C4H3FN2O2 taking 552 as the specific
C. To a volume of the injection containing 50 mg of Fluorouracil
add 1 ml of bromine water; the colour of bromine is discharged.
Storage. Store protected from light in single dose containers
Tests at a temperature not exceeding 30°. The injection should not
pH (2.4.24). 8.5 to 9.5. be allowed to freeze.
516
IP 2007 FLUTICASONE PROPIONATE
Labelling. The label states (1) the strength in terms of the Chromatographic system
equivalent amount of Fluorouracil in a suitable dose-volume; – a stainless steel column 25 cm x 4.6 mm, packed with
(2) that, if separation has occurred, the injection should be octadecylsilane bonded to porous silica (5 µm),
heated to 60°, shaken vigorously and allowed to cool to body – column temperature 40º,
temperature prior to use. – mobile phase: A. 0.05 per cent v/v orthophosphoric
acid and 3 per cent v/v methanol in acetonitrile.
B. 0.05 per cent v/v orthophosphoric
acid and 3 per cent v/v methanol in water.
Fluticasone Propionate – flow rate. 1 ml per minute,
F O below,
O – spectrophotometer set at 239 nm,
S
H3C CH3 – a 50 µl loop injector.
HO O
H3C H CH3 Time Mobile phase A Mobile phase B
(min.) (per cent v/v) (per cent v/v)
F H 0 43 57
O 40 55 45
F 60 90 10
C25H31F3O5S Mol. Wt. 500.6 70 90 10

75 43 57
Fluticasone Propionate is 5-fluoromethyl-6α,9α-difluoro-
11β-hydroxy-16α-methyl-17α-propionyloxy-3-oxoandrosta- 85 43 57
1,4-diene-17β-carbothioate. Inject the reference solution. The test is not valid unless the
Fluticasone Propionate contains not less than 96.0 per cent tailing factor is not more than 2.0 and the column efficiency in
and not more than 102.0 per cent of fluticasone, C25H31F3O5S, not less than 20,000 theoretical plates.
calculated on the anhydrous basis. Inject the test solution. Any individual impurity is not more
Description. A white or almost white powder. than 0.5 per cent and sum of all the impurities is not more than
2.0 per cent.
Identification Water (2.3.43). Not more than 0.5 per cent, determined on 0.25
A. Determine by infrared absorption spectrophotometry (2.4.6). g, using as solvent a mixture of equal volumes of chloroform
Compare the spectrum with that obtained with fluticasone and methanol.
propionate RS or with the reference spectrum of fluticasone Assay. Determine by liquid chromatography (2.4.14).
propionate.
B. In the Assay, the principal peak in the chromatogram examination in 100.0 ml of the mobile phase. Dilute 10.0 ml of
obtained with the test solution corresponds to the principal this solution to 100.0 ml with the mobile phase.
peak in the chromatogram obtained with the reference solution.
Tests fluticasone propionate RS in the mobile phase.
Specific optical rotation (2.4.22). + 32.0º to + 36.0º, determined Chromatographic system

in a 0.5 per cent w/v solution in dichloromethane. – a stainless steel column 25 cm x 4.6 mm, packed with
Related substances. Determine by liquid chromatography – column temperature 40º,
(2.4.14). – mobile phase: a mixture of 15 volumes of acetonitrile,
Solvent mixture. 50 volumes of mobile phase A and 50 volumes 35 volumes of a buffer solution prepared by dissolving
of mobile phase B. 1.15 g of ammonium dihydrogen phosphate in 1000 ml
of water and adjusting the pH to 3.5 with
Test solution. Dissolve 20 mg of the substance under orthophosphoric acid, and 50 volumes of methanol,
examination in 100 ml of the solvent mixture. – flow rate. 1.5 ml per minute,
Reference solution. A 0.02 per cent w/v solution fluticasone – spectrophotometer set at 239 nm,
propionate RS in the solvent mixture. – a 20 µl loop injector.
517
FLUTICASONE PROPIONATE INHALATION IP 2007
Inject the reference solution. The test is not valid unless relative – mobile phase: a mixture of 40 volumes of a buffer solution
standard deviation for replicate injections is not more than 2.0 prepared by dissolving 1.3 g of diammonium hydrogen
per cent. orthophosphate in 1000 ml of water and adjusting the
Inject the test solution and the reference solution. pH to 7.0 with orthophosphoric acid, and 60 volumes
of acetonitrile.
Calculate the content of C25H31F3O5S. – flow rate. 2 ml per minute,
Storage. Store protected from light. – spectrophotometer set at 238 nm,
– inject 200 µl.
Inject the reference solution (b). The test is not valid unless
the column efficiency is not less than 1500 theoretical plates,
Fluticasone Propionate Inhalation the tailing factor is not more than 2.0 and the relative standard
Fluticasone Propionate Inhalation is a suspension of microfine
Fluticasone Propionate in a suitable liquid filled in a suitable Inject the test solution and reference solution (b).
pressurized container. It may contain suitable pharmaceutical Calculate the content of C25H31F3O5S in the solution and the
aids such as surfactants, stabilizing agents. content of C25H31F3O5S delivered per actuation of the valve.
Fluticasone Propionate Inhalation delivers not less than 80.0 Determine the content of active ingredient a second and third
per cent and not more than 120.0 per cent of the stated amount time by repeating the procedure on the middle ten and on the
of fluticasone propionate, C25H31F3O5S, per inhalation by last ten successive combined actuations of the valve. For
actuation of the valve. each of the three determinations the average content of
C25H31F3O5S delivered per actuation of the valve meets the
Identification requirements.
In the Assay, the principal peak in the chromatogram obtained Storage. Store protected from moisture at a temperature not
with the test solution corresponds to the principal peak in the exceeding 30º.
chromatogram obtained with reference solution (b). Labelling. The label states the amount of active ingredient
delivered per inhalation.
Tests
Other tests. Complies with the tests stated under Inhalation
Preparations (Pressurised Metered-dose Preparations). Fluticasone Propionate Powder for
Follow the procedure described under Assay with suitable Inhalation
dilution of the reference solution wherever the amount of
Fluticasone Propionate Powder for Inhalation consists of
active substance is to be determined in any test.
Fluticasone propionate in microfine powder either alone or
Assay. Carry out the test for Content of active ingredient admixed with Lactose in a pre-metered unit for use in a suitable
delivered per actuation stated under Inhalation Preparations powder inhaler.
(Pressurised Metered-dose Preparations). Fluticasone Propionate Powder for Inhalation contains not
Determine by liquid chromatography (2.4.14). less than 90.0 per cent and not more than 125.0 per cent of the
stated amount of fluticasone propionate, C25H31F3O5S per unit
Test solution. Prepare using the mobile phase as described
dose.
under the test for Content of active ingredient delivered per
actuation stated under Inhalation Preparations (Pressurised Identification
Metered-dose Preparations).
Reference solution (a). A 0.05 per cent w/v solution of with the test solution corresponds to the peak in the
fluticasone propionate RS in acetonitrile. chromatogram obtained with the reference solution.
Reference solution (b). Dilute reference solution (a) with the
mobile phase to obtain a solution containing 25 µg of Tests
fluticasone propionate per ml Other tests. Complies with the tests stated under Inhalation
Chromatographic system Preparations (Powders for Inhalation).
– a stainless steel column 15 cm x 4.6 mm, packed with Follow the procedure described under Assay with suitable
octadecylsilyl silica gel (5 mm), dilution of the reference solution wherever the amount of
– column temperature 40º, active substance is to be determined in any test.
518
IP 2007 FLUPHENAZINE DECANOATE
Assay. Determine by liquid chromatography (2.4.14). Fluphenazine Decanoate is 2-{4-[3-(2-

Test solution. To 10 intact capsules add 10 ml of water and mix trifluoromethylphenothiazin-10-yl)propyl]piperazin-1-
with the aid of ultrasound till the shells get dispersed. Add 60 yl}ethyl decanoate.
ml of the mobile phase and continue mixing with the aid of Fluphenazine Decanoate contains not less than 98.5 per cent
ultrasound for 10 minutes with intermittent shaking. Add and not more than 101.5 per cent of C32H44F3N3O2S, calculated
sufficient of the mobile phase to produce 100.0 ml. Dilute on the dried basis.
suitably, if required, to get a solution containing 25 µg of
Description. A pale yellow, viscous liquid or yellow, crystalline,
Fluticasone Propionate per ml.
oily solid; odour, faint and ester-like.
Reference solution. A solution containing 0.5 mg of
fluticasone propionate per ml prepared by dissolving 10 mg of Identification
fluticasone propionate RS in 10 ml acetonitrile and adding
sufficient of the mobile phase to produce 20 ml and furthur A. Determine by infrared absorption spectrophotometry (2.4.6).
dilute with mobile phase to obtain a solution containing 25 µg Compare the spectrum with that obtained with fluphenazine
of Fluticasone Propionate per ml. . decanoate RS or with the reference spectrum of fluphenazine
decanoate.
– a stainless steel column 15 cm x 4.6 mm, packed with B. When examined in the range 230 nm to 360 nm (2.4.7), a
octadecylsilyl silica gel (5 µm), 0.001 per cent w/v solution in ethanol shows an absorption
– column temperature 40º, maximum at about 261 nm and a less well-defined maximum at
– mobile phase: a mixture of 40 volumes of a buffer solution about 310 nm; absorbance at about 261 nm, about 0.60.
prepared by dissolving 1.3 g of diammonium hydrogen C. Determine in subdued light by thin-layer chromatography
orthophosphate in 1000 ml of water and adjusting the (2.4.17), coating the plate with silica gel GF254. Impregnate
pH to 7.0 with orthophosphoric acid and 60 volumes of the dry plate by placing it in a tank containing a 5 per cent
acetonitrile, v/v solution of n-tetradecane in n-hexane, allowing the
– flow rate. 2 ml per minute, impregnating solvent to ascend to the top and allowing to
– spectrophotometer set at 238 nm, dry.
Mobile phase. Methanol (90 per cent).
examination in 10 ml of ethanol (95 per cent).
the tailing factor is not more than 2.0 and the relative standard
deviation for replicate injections is not more than 2.0 per cent. Reference solution. A 2.0 per cent w/v solution of
Inject the test solution and the reference solution. fluphenazine decanoate RS in ethanol (95 per cent).
Calculate the content of C25H31F3O5S per unit. Apply to the plate 1 µl of each solution. After development,
dry the plate dry in air and examine in ultraviolet light at
Storage. Store protected from moisture, at temperature not 254 nm. The principal spot in the chromatogram obtained with
exceeding 30º. the test solution corresponds to that in the chromatogram
Labelling. The label states the quantity of active ingredient obtained with the reference solution.
per pre-metered unit.
D. Dissolve 5 mg in 2 ml of sulphuric acid and allow to stand
for 5 minutes; a reddish-brown colour is produced.
Fluphenazine Decanoate Tests

Fluphenazine Decanoate Ester Related substances. Determine in subdued light by thin-layer
N F254.
N O (CH2 )8 Mobile phase. A mixture of 80 volumes of acetone, 30 volumes
CH3 of cyclohexane and 5 volumes of strong ammonia solution.
N CF3 O
S
C32H44F3N3O2S Mol. Wt. 591.8 substance under examination in methanol.
519
FLUPHENAZINE DECANOATE INJECTION IP 2007
Apply to the plate 20 µl of each solution. After development, ultraviolet light at 254 nm. The principal spot in the
dry the plate in air and examine in ultraviolet light at 254 nm. chromatogram obtained with the test solution corresponds to
Spray the plate with sulphuric acid (50 per cent v/v) and that in the chromatogram obtained with the reference solution.
examine in daylight. By both methods of visualisation, any
B. Shake a volume of the injection containing 5 mg of
Fluphenazine Decanoate with 1 ml of a 1 per cent w/v solution
obtained with the reference solution. of sucrose in hydrochloric acid and allow to stand for 5
minutes; a red colour is produced in the acid layer.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Tests
on 1.0 g by drying in an oven at 60° at a pressure not exceeding Related substances. Determine in subdued light by thin-layer
0.7 kPa for 3 hours. chromatography (2.4.17),coating the plate with silica gel
Assay. Weigh accurately about 0.6 g and dissolve in 50 ml of GF254.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Mobile phase. Chloroform for the first development and a
acid, using crystal violet solution as indicator. Carry out a mixture of 80 volumes of acetone, 30 volumes of cyclohexane
blank titration. and 5 volumes of strong ammonia solution for the second
1 ml of 0.1 M perchloric acid is equivalent to 0.02959 g of development.
C32H44F3N3O2S. Test solution. Dilute a suitable volume of the injection with
Storage. Store protected from light. ethanol (95 per cent) to produce a solution containing 2.5 mg
of Flufenazine Decanoate per ml.
fluphenazine hydrochloride RS in methanol.
Fluphenazine Decanoate Injection Apply to the bottom right-hand corner of the plate 10 µl of the
Fluphenazine Decanoate Injection is a sterile solution of test solution. After development, dry the plate in air, turn it
Fluphenazine Decanoate in Sesame Oil. through 90° in a clockwise direction. Apply to the bottom
right-hand corner of the plate, to the right of the solvent front
Fluphenazine Decanoate Injection contains not less than 90.0 of the first development, 1 µl of the reference solution. After
per cent and not more than 110.0 per cent of the stated amount the second development, dry the plate in air and examine in
of fluphenazine decanoate, C32H44F3N3O2S. ultraviolet light at 254 nm. Spray the plate with sulphuric acid
(50 per cent v/v). By both methods of visualisation, any
Identification secondary spot in the chromatogram obtained with the test
A.Determine in subdued light by thin-layer chromatography solution is not more intense than the spot in the chromatogram
(2.4.17), coating the plate with silica gel GF254. obtained with the reference solution.
Mobile phase. Chloroform for the first development and Other tests. Complies with the tests stated under Parenteral
methanol (90 per cent) for the second development. Preparations (Injections).
Test solution. Dilute a suitable volume of the injection with Assay. Carry out the following procedure protected from
ethanol (95 per cent) to produce a solution containing 2.5 mg light.
of Flufenazine Decanoate per ml. Measure accurately a volume of the injection containing about
Reference solution. A 0.25 per cent w/v solution of 0.25 g of Fluphenazine Decanoate and dilute with 75 ml of
fluphenazine decanoate RS in ethanol (95 per cent). anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
acid, using crystal violet solution as indicator. Carry out a
Apply to the bottom right-hand corner of the plate 10 µl of the blank titration.
test solution. After development, dry the plate in air, turn the
plate through 90° in a clockwise direction, impregnate the 1 ml of 0.1 M perchloric acid is equivalent to 0.02959 g of
coating with a 5 per cent v/v solution of n-tetradecane in n- C32H44F3N3O2S.
hexane and allow it to dry in air. Apply to the bottom right- Storage. Store protected from light.
hand corner of the plate, to the right of the solvent front of the
first development, 10 µl of the reference solution. After the Labelling. The label states that the injection is for intramuscular
second development, dry the plate in air and examine in injection only.
520
IP 2007 FLUPHENAZINE HYDROCHLORIDE
Fluphenazine Hydrochloride Reference solution. A 0.2 per cent w/v solution of

OH Apply to the plate 2 µl of each solution. After development,
N
dry the plate in air, examine in ultraviolet light at 365 nm and
N observe the fluorescence produced after about 2 minutes.
Heat the plate at 120° for 20 minutes, cool, spray with ethanolic
N CF3 ,2HCl sulphuric acid (20 per cent) and observe the colour produced.
S solution corresponds to that in the chromatogram obtained
C22H26F3N3OS,2HCl Mol. Wt. 510.5
D. Mix about 5 mg with 45 mg of heavy magnesium oxide and
Fluphenazine Hydrochloride is 2-{4-[3-(2-
ignite in a crucible until an almost white residue is obtained
trifluoromethylphenothiazin-10-yl)propyl]piperazin-1-
(usually less than 5 minutes). Allow to cool, add 1 ml of water,
yl}ethanol dihydrochloride.
0.05 ml of dilute phenolphthalein solution and about 1 ml of
Fluphenazine Hydrochloride contains not less than 98.5 per 2 M hydrochloric acid to render the solution colourless and
cent and not more than 101.0 per cent w/v of filter. To a freshly prepared mixture of 0.1 ml of alizarin red S
C22H26F3N3OS,2HCl, calculated on the dried basis. solution and 0.1 ml of zirconyl nitrate solution, add 1 ml of
Description. A white or almost white, crystalline powder. the filtrate. Mix, allow to stand for 5 minutes and examine the
colour of the solution as well as of a blank prepared in the
Identification same manner. The colour of the test solution is yellow and
that of the blank is red.
Tests B, C and D may be omitted if tests A and E are carried E. Gives the reaction of chlorides (2.3.1).
out.
Tests
A. Dissolve 0.1 g in 10 ml of water, make alkaline with 1 M
sodium hydroxide, extract with 5 ml of chloroform, filter pH (2.4.24). 1.9 to 2.3, determined in a 5.0 per cent w/v solution
through anhydrous sodium sulphate and evaporate the in carbon dioxide-free water.
solvent in a current of nitrogen. The oily residue complies Related substances. Determine in subdued light by thin-layer
with the following test. chromatography (2.4.17), coating the plate with silica gel
Determine by infrared absorption spectrophotometry (2.4.6). GF254.
Compare the spectrum with that obtained with fluphenazine Mobile phase. A mixture of 80 volumes of acetone, 30 volumes
hydrochloride RS treated in the same manner or with the of cyclohexane and 5 volumes of strong ammonia solution.
reference spectrum of fluphenazine.
B. When examined in the range 230 nm to 360 nm (2.4.7), a examination in 10 ml of 0.1 M methanolic sodium hydroxide.
maximum at about 258 nm and a less well-defined maximum at Reference solution (a). Dilute 1 ml of the test solution to100
about 310 nm; absorbance at about 258 nm, between 0.63 and ml with 0.1 M methanolic sodium hydroxide.
0.70. Reference solution (b). Dilute 5 ml of reference solution (a)
C. Determine in subdued light by thin-layer chromatography to 10 ml with 0.1 M methanol sodium hydroxide.
(2.4.17), coating the plate with kieselguhr G. Impregnate the Apply to the plate 10 µl of each solution. After development,
dry plate by placing in a tank containing a shallow layer of a dry the plate in air and examine in ultraviolet light at 254 nm.
mixture of 36 volumes of acetone, 3 volumes of formamide Any secondary spot in the chromatogram obtained with the
and 1 volume of 2-phenoxyethanol. Allow the impregnating test solution is not more intense than the spot in the
solvent to ascend to the top, remove the plate from the tank chromatogram obtained with reference solution (a) and not
and use it immediately. more than one such spot is more intense than the spot in the
Mobile phase. A mixture of 100 volumes light petroleum (40° chromatogram obtained with reference solution (b). Ignore
to 60°) saturated with 2-phenoxyethanol and 2 volumes of any spot remaining on the line of application.
diethylamine. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Test solution. Dissolve 0.2 g of the substance under Loss on drying (2.4.19). Not more than 1.0 per cent, determined
examination in 100 ml of methanol. on 1.0 g by drying in an oven at 105° for 3 hours.
521
FLUPHENAZINE HYDROCHLORIDE INJECTION IP 2007
Assay. Weigh accurately about 0.25 g and dissolve in a mixture Related substances. Determine in subdued light by thin-layer
of 10 ml of anhydrous formic acid and 40 ml of acetic chromatography (2.4.17), coating the plate with silica gel
anhydride. Titrate with 0.1 M perchloric acid, determining GF254.
the end-point potentiometrically (2.4.25). Carry out a blank Mobile phase. A mixture of 80 volumes of acetone, 30 volumes
titration. of cyclohexane and 5 volumes of strong ammonia solution.
1 ml of 0.1 M perchloric acid is equivalent to 0.02552 g of Test solution. Dilute a quantity of the injection containing 20
C22H26F3N3OS,2HCl. mg of Fluphenazine Hydrochloride with sufficient 0.1 M
Storage. Store protected from light. methanolic sodium hydroxide to make 10 ml.
50 volumes with 0.1 M methanolic sodium hydroxide.
Fluphenazine Hydrochloride Injection Reference solution (b). Dilute 1 volume of the test solution to
100 volumes with 0.1 M methanolic sodium hydroxide.
Fluphenazine Hydrochloride Injection is a sterile solution of
Fluphenazine Hydrochloride in Water for Injection. Apply to the plate 50 µl of the test solution and 25 µl of
reference solutions (a) and (b). After development, dry the
Fluphenazine Hydrochloride Injection contains not less than
plate in air and examine in ultraviolet light at 254 nm. Any
amount of fluphenazine hydrochloride, C22H26F3N3OS,2HCl.
Description. A clear, colourless solution. obtained with reference solution (a) and not more than two
such spots are more intense than the spot in the chromatogram
Identification obtained with reference solution (b). Ignore any spot remaining
on the line of application.
A. Determine in subdued light by thin-layer chromatography
(2.4.17), coating the plate with kieselguhr G. Impregnate the Other tests. Complies with the tests stated under Parenteral
dry plate by placing in a tank containing a shallow layer of a Preparations (Injections).
mixture of 36 volumes of acetone, 3 volumes of formamide Assay. Carry out the following procedure protected from
and 1 volume of 2-phenoxyethanol. Allow the impregnating light.
solvent to ascend to the top, remove the plate from the tank
and use it immediately. To an accurately measured quantity of the injection containing
about 5 mg of Fluphenazine Hydrochloride add a mixture of 1
Mobile phase. A mixture of 100 volumes light petroleum (40° volume of 1 M hydrochloric acid and 99 volumes of ethanol
to 60°) saturated with 2-phenoxyethanol and 2 volumes of (90 per cent) to produce 50.0 ml. Dilute 10.0 ml of this solution
diethylamine. to 100.0 ml with the acid-ethanol mixture and measure the
Test solution. Use a quantity of the injection containing 2 mg absorbance of the resulting solution at the maximum at about
of Fluphenazine Hydrochloride and dilute it to 1 ml with 258 nm (2.4.7). Calculate the content of C22H26F3N3OS,2HCl
methanol. taking 620 as the specific absorbance at 258 nm.
Reference solution. A 0.2 per cent w/v solution of Storage. Store protected from light.
After development, dry the plate in air, examine in ultraviolet
light at 365 nm and observe the fluorescence produced after Fluphenazine Tablets
about 2 minutes. Heat the plate at 120° for 20 minutes, cool,
Fluphenazine Hydrochloride Tablets
spray with ethanolic sulphuric acid (20 per cent) and observe
the colour produced. The principal spot in the chromatogram Fluphenazine Tablets contain not less than 90.0 per cent and
obtained with the test solution corresponds to that in the not more than 110.0 per cent of the stated amount of
chromatogram obtained with the reference solution. fluphenazine hydrochloride, C22H26F3N3OS,2HCl. The tablets
are coated.
B. To a quantity of the injection containing 5 mg of
Fluphenazine Hydrochloride add 2 ml of sulphuric acid and Identification
allow to stand for 5 minutes; an orange colour is produced.
A. Determine in subdued light by thin-layer chromatography
Tests (2.4.17), coating the plate with kieselguhr G. Impregnate the
dry plate by placing in a tank containing a shallow layer of a
pH (2.4.24). 4.8 to 5.2. mixture of 36 volumes of acetone, 3 volumes of formamide
522
IP 2007 FLURBIPROFEN
and 1 volume of 2-phenoxyethanol. Allow the impregnating Apply to the plate 50 µl of the test solution and 25 µl of
solvent to ascend to the top, remove the plate from the tank reference solutions (a) and (b). After development, dry the
and use it immediately. plate in air and examine in ultraviolet light at 254 nm. Any
Mobile phase. A mixture of 100 volumes light petroleum (40° secondary spot in the chromatogram obtained with the test
to 60°) saturated with 2-phenoxyethanol and 2 volumes of solution is not more intense than the spot in the chromatogram
diethylamine. obtained with reference solution (a) and not more than two
such spots is more intense than the spot in the chromatogram
Test solution. Shake a quantity of the powdered tablets with obtained with reference solution (b). Ignore any spot remaining
sufficient methanol to produce a solution containing 0.2 per on the line of application.
cent w/v of Fluphenazine Hydrochloride, centrifuge and use
the supernatant liquid.
Tablets.
Carry out the procedure protected from light.
Powder 1 tablet and dissolve the powder as completely as
After development, dry the plate in air, examine in ultraviolet
possible in a mixture of 99 volumes of ethanol (80 per cent)
light at 365 nm and observe the fluorescence produced after
and 1 volume of 1 M hydrochloric acid. Add sufficient of the
about 2 minutes. Heat the plate at 120° for 20 minutes, cool,
acid-ethanol mixture to produce 100.0 ml and filter. Dilute
spray with ethanolic sulphuric acid (20 per cent) and observe
suitably, if necessary with the acid-ethanol mixture and
the colour produced. The principal spot in the chromatogram
C22H26F3N3OS,2HCl taking 620 as the specific absorbance at
B. Extract a quantity of the powdered tablets containing 5 mg 258 nm.
of Fluphenazine Hydrochloride with 5 ml of acetone, filter and
evaporate the filtrate to dryness. Add 2 ml of sulphuric acid
to the residue and allow to stand for 5 minutes; an orange Assay. Carry out the following procedure protected from
colour is produced. light.
C. Extract a quantity of the powdered tablets containing 10 mg Weigh and powder 20 tablets. Weigh accurately a quantity of
of Fluphenazine Hydrochloride with 10 ml of ethanol the powder containing about 5 mg of Fluphenazine
containing 0.2 per cent v/v of strong ammonia solution and Hydrochloride, dissolve as completely as possible in a mixture
evaporate the extract to dryness. Heat 0.5 ml of chromic- of 99 volumes of ethanol (80 per cent) and 1 volume of 1 M
sulphuric acid mixture in a small test-tube in a water-bath for hydrochloric acid, add sufficient of the acid-ethanol mixture
5 minutes; the solution wets the sides of the tube readily and to produce 100.0 ml and filter. Dilute 10.0 ml of the filtrate to
there is no greasiness. Add 2 or 3 mg of the residue and again 50.0 ml with the acid-ethanol mixture and measure the
heat in a water-bath for 5 minutes; the solution does not wet absorbance of the resulting solution at the maximum at about
the sides of the tube and does not pour easily from the tube. 258 nm (2.4.7). Calculate the content of C22H26F3N3OS,2HCl
Tests Storage. Store protected from light.
Related substances. Determine in subdued light by thin-layer
GF254. Flurbiprofen
of cyclohexane and 5 volumes of strong ammonia solution. CH3
F
Test solution. Remove the coating from a suitable quantity of COOH
tablets; shake a quantity of the powdered tablet cores
containing 20 mg of Fluphenazine Hydrochloride with 10 ml of
0.1 M methanolic sodium hydroxide for 5 minutes, centrifuge
and use the supernatant liquid.
Reference solution (a). Dilute 1 volume of the test solution to C15H13FO2 Mol. Wt. 244.3
50 volumes with 0.1 M methanolic sodium hydroxide. Flurbiprofen is (RS)-2-(2-fluorobiphenyl-4-yl)propionic acid.
Reference solution (b). Dilute 1 volume of the test solution to Flurbiprofen contains not less than 99.0 per cent and not more
100 volumes with 0.1 M methanolic sodium hydroxide. than 100.5 per cent of C15H13FO2, calculated on the dried basis.
523
FLURBIPROFEN TABLETS IP 2007
Description. A white or almost white, crystalline powder. The test is not valid unless a peak due to 2-(biphenyl-4-yl)
propionic acid appears immediately before the principal peak
Identification in the chromatogram obtained with test solution (b) and the
height of the trough separating the two peaks is less than 4
per cent of the full-scale deflection on the chart paper.
Compare the spectrum with that obtained with flurbiprofen
RS or with the reference spectrum of flurbiprofen. Heavy metals (2.3.13). 2.0 g complies with the limit test for
0.001 per cent w/v solution in 0.1 M sodium hydroxide shows Sulphated ash (2.3.18). Not more than 0.1 per cent.
an absorption maximum only at about 247 nm; absorbance at Loss on drying. Not more than 0.5 per cent, determined on 1.0
about 247 nm, about 0.8. g by drying in an oven at 60° at a pressure not exceeding 0.7
C. Heat 0.5 ml of chromic-sulphuric acid mixture in a small kPa.
test-tube in a water-bath for 5 minutes; the solution wets the Assay. Weigh accurately about 0.5 g, dissolve in 100 ml of
sides of the tube readily and there is no greasiness. Add 2 or ethanol (95 per cent) previously neutralised to
3 mg of the substance under examination and heat in a water- phenolphthalein solution and titrate with 0.1 M sodium
bath for 5 minutes; the solution does not wet the sides of the hydroxide using phenolphthalein solution as indicator.
tube and does not pour easily from the tube.
Tests C15H13FO2.

(2.4.17). Flurbiprofen Tablets
Solvent mixture. A mixture of 45 volumes of acetonitrile and
Flurbiprofen Tablets contain not less than 92.5 per cent and
55 volumes of water.
Test solution (a). Dissolve 0.2 g of the substance under flurbiprofen, C15H13FO2. The tablets are coated.
Identification
Test solution (b). Dissolve 0.2 g of the substance under
examination and 1 mg of 2-(biphenyl- 4-yl) propionic acid RS Extract a quantity of the powdered tablets containing 0.5 g of
in 100 ml of the same solvent mixture. Flurbiprofen with 25 ml of acetone, filter, evaporate the filtrate
to dryness with the aid of a current of air without heating and
Reference solution. A 0.001 per cent w/v solution of 2- dry at 60° at a pressure of 2 kPa. The residue complies with the
(biphenyl-4-yl) propionic acid RS in the same solvent mixture. following tests.
Chromatographic system A. Determine by infrared absorption spectrophotometry (2.4.6).
– a stainless steel column 15 cm x 3.9 mm, packed with Compare the spectrum with that obtained with flurbiprofen
octadecylsilyl silica gel (5 µm), RS or with the reference spectrum of flurbiprofen.
volumes of acetonitrile and 5 volumes of glacial acetic B. Heat 0.5 ml of chromic-sulphuric acid mixture in a small
acid, test-tube in a water-bath for 5 minutes; the solution wets the
– flow rate. 1 ml per minute, sides of the tube readily and there is no greasiness. Add 2 or
– spectrophotometer set at 254 nm, 3 mg of the substance under examination and heat in a water-
– a 20 µl loop injector. bath for 5 minutes; the solution does not wet the sides of the
tube and does not pour easily from the tube.
Adjust the sensitivity of the instrument so that with the
reference solution the height of the peak due to 2-(biphenyl- Tests
4-yl)propionic acid is about 40 per cent of the full-scale
deflection on the recorder.
(2.4.17).
In the chromatogram obtained with test solution (a) the area
Test solution. Disperse a quantity of the powdered tablets
of any secondary peak is not greater than the area of the peak
containing 0.5 g of Flurbiprofen in 50 ml of water, add 200 ml
in the chromatogram obtained with the reference solution and
of acetonitrile, mix and centrifuge. Use the supernatant liquid.
the sum of the areas of any such peaks is not greater than
twice the area of the peak in the chromatogram obtained with Reference solution. A 0.001 per cent w/v solution of 2-
the reference solution. (biphenyl-4-yl)propionic acid RS in the test solution.
524
IP 2007 FOLIC ACID
Chromatographic system Description. A yellow to yellowish-orange, crystalline powder;

– a stainless steel column 15 cm x 3.9 mm, packed with odourless or almost odourless.
– mobile phase: a mixture of 60 volumes of water, Identification
35 volumes of acetonitrile and 5 volumes of glacial
acetic acid, A. When examined in the range 230 nm to 380 nm (2.3.13), a
– flow rate. 1 ml per minute, 0.001 per cent w/v solution in 0.1 M sodium hydroxide shows
– spectrophotometer set at 254 nm, three absorption maxima, at about 256 nm, 283 nm and 365 nm;
– a 20 µl loop injector. absorbance at about 256 nm, about 0.59, at about 283 nm,
about 0.575 and at about 365 nm, about 0.206.
Adjust the sensitivity of the instrument so that with the
reference solution the height of the peak due to 2-(biphenyl- B. Determine by thin-layer chromatography (2.4.17), coating
4-yl) propionic acid is about 40 per cent of the full-scale the plate with silica gel G.
deflection on the recorder. Mobile phase. A mixture of 60 volumes of ethanol (95 per
In the chromatogram obtained with the test solution the area cent), 20 volumes of strong ammonia solution and 20 volumes
of any secondary peak is not greater than the area of the peak of 1-propanol.
in the chromatogram obtained with the reference solution and Test solution. Dissolve 50 mg of the substance under
the sum of the areas of any such peaks is not greater than examination in 100 ml of a mixture of 9 volumes of methanol
twice the area of the peak in the chromatogram obtained with and 2 volumes of strong ammonia solution.
the reference solution.
Reference solution. A 0.05 per cent w/v solution of folic acid
Other tests. Comply with the tests stated under Tablets. RS in a mixture of 9 volumes of methanol and 2 volumes of
Assay. Weigh and powder 20 tablets. Weigh accurately a strong ammonia solution.
quantity of the powder containing about 0.1 g of Flurbiprofen, Apply to the plate 2 µl of each solution. After development,
shake with 60 ml of 0.1 M sodium hydroxide for 5 minutes, dry the plate in air and examine in ultraviolet light at 365 nm.
dilute to 100.0 ml with 0.1 M sodium hydroxide, filter if The principal spot in the chromatogram obtained with the test
necessary and dilute 10.0 ml of the filtrate to 100.0 ml with the solution corresponds to that in the chromatogram obtained
same solvent. Further dilute 10.0 ml to 100.0 ml with the same with the reference solution.
solvent and measure the absorbance of the resulting solution
at the maximum at about 247 nm (2.4.7). Calculate the content Tests
of C15H13FO2 taking 802 as the specific absorbance at 247 nm.
Specific optical rotation (2.4.22). About +20°, determined in a
1.0 per cent w/v solution in 0.1 M sodium hydroxide.
Free amines. The absorbance (2.4.7) of the unreduced solution
Folic Acid as determined in the Assay is not more than one-sixth of the
absorbance of the reduced solution.
Pteroylglutamic Acid Sulphated ash (2.3.18). Not more than 0.5 per cent.
H
H2N N N Assay. Weigh accurately about 50 mg, dissolve in 50 ml of
H 0.1 M sodium hydroxide and add sufficient 0.1 M sodium
N N
N hydroxide to produce 100.0 ml (solution A). To 3.0 ml of solution
H
O N COOH A add 20 ml of 2 M hydrochloric acid and dilute to 100.0 ml
with water. To 50 ml add 0.5 g of zinc powder, allow to stand
O COOH protected from light for 20 minutes with frequent shaking and
filter. Dilute 10.0 ml of the filtrate to 25 ml with water, add 5 ml
C19H19N7O6 Mol. Wt. 441.4 of 2 M hydrochloric acid and 5 ml of a 0.1 per cent w/v solution
of sodium nitrite, mix and allow to stand for 2 minutes. Add ml
Folic Acid is (2S)-[4-[(2-amino-4-hydroroxypteridin-6-yl) of a 0.5 per cent w/v solution of ammonium sulphamate, mix
methylamino]benzamido]glutamic acid. and allow to stand for 2 minutes. Add 5 ml of a 0.1 per cent
Folic acid contains not less than 95.0 per cent and not more w/v solution of N-(1-naphthyl)ethylenediamine
than 102.0 per cent of C19H19N7O6, calculated on the anhydrous dihydrochloride, mix and allow to stand for 10 minutes. Add
basis. sufficient water to produce 50.0 ml and measure the
525
FOLIC ACID TABLETS IP 2007
absorbance of the resulting solution at about 550 nm (2.4.7), Tests

using as the blank a solution prepared in a similar manner but
using 25 ml of water and beginning at the words “add 5 ml of Hydrolysis products. Determine by liquid chromatography
2 M hydrochloric acid...”. To a further 30.0 ml of solution A (2.4.14).
add 20 ml of 2 M hydrochloric acid and sufficient water to Protect the solutions from light.
produce 100.0 ml. Mix 10.0 ml of this solution with 15 ml of Test solution. Shake a quantity of the powdered tablets
water and carry out the operations described above beginning containing 5.0 mg of Folic Acid with 50 ml of the mobile phase,
at the words “add 5 ml of 2 M hydrochloric acid...”. Subtract centrifuge and use the supernatant liquid.
one-tenth of the absorbance of the unreduced solution from
that of the reduced solution and from the result calculate the Reference solution. A solution containing 0.00005 per cent
amount of C19H19N7O6, using the result obtained by repeating w/v of 4-aminobenzoic acid and 0.0002 per cent w/v of N-(4-
the operation using folic acid RS instead of the substance aminobenzoyl)-L-glutamic acid in the mobile phase.
under examination. Chromatographic system
Storage. Store protected from light. – a stainless steel column 20 cm x 4.6 mm, packed with
octadecylsilyl silica gel (5 µm) (such as Spherisorb ODS
1),
– mobile phase: 0.05 M potassium dihydrogen phosphate,
Folic Acid Tablets adjusted to pH 5.5 with 5 M sodium hydroxide,
Folic Acid Tablets contain not less than 90.0 per cent and not – flow rate. 2 ml per minute,
more than 115.0 per cent of the stated amount of folic acid, – spectrophotometer set at 269 nm,
C19H19N7O6. – a 20 µl loop injector.
In the chromatogram obtained with the reference solution two
Identification peaks due to N-(4-aminobenzoyl)-L-glutamic acid and 4-
aminobenzoic acid are obtained in the order of their
emergence.The areas of the peaks due to 4-aminobenzoic-
acid and N-(4-aminobenzoyl)-L-glutamic acid in the
Mobile phase. A mixture of 60 volumes of ethanol (95 per chromatogram obtained with the reference solution are greater
cent), 20 volumes of strong ammonia solution and 20 volumes than the areas of any corresponding peaks in the chromatogram
of 1-propanol. obtained with the test solution. The test is not valid unless
Test solution. Extract a quantity of the powdered tablets the resolution between the two peaks in the chromatogram
containing 0.5 mg of Folic Acid with 1 ml of a mixture of 1 obtained with the reference solution is greater than 3.
volume of strong ammonia solution and 9 volumes of Uniformity of content. Comply with the test stated under
methanol, centrifuge and use the supernatant liquid. Tablets.
Reference solution. A 0.05 per cent w/v solution of folic acid Test solution. Shake 1 tablet with 5.0 ml of 0.1 M sodium
RS in a mixture of 9 volumes of methanol and 2 volumes of hydroxide, add sufficient mobile phase to produce a solution
strong ammonia solution. containing 0.001 per cent w/v of Folic Acid, centrifuge and
use the supernatant liquid.
dry the plate in air and examine in ultraviolet light at 365 nm. Reference solution. Add 1.0 ml of 0.5 M hydrochloric acid to
The principal spot in the chromatogram obtained with the test 5.0 ml of a 0.002 per cent w/v solution of folic acid RS in 0.1 M
solution corresponds to that in the chromatogram obtained sodium hydroxide and dilute to 10.0 ml with the mobile phase.
with the reference solution. Chromatographic system
B. Dissolve a quantity of the powdered tablets containing – a stainless steel column 20 cm x 4.6 mm, packed with
5 mg of Folic Acid as completely as possible in 5 ml of 0.1 M octadecylsilyl silica gel (5 µm) (such as Spherisorb ODS
sodium hydroxide and filter. To the filtrate add 45 ml of 0.5 M 1),
hydrochloric acid and 5 g of zinc powder and shake for 30 – mobile phase: 0.05 M potassium dihydrogen phosphate,
minutes. To 5 ml of the reduced solution, add 2 ml of a 0.1 per adjusted to pH 5.5 with 5 M sodium hydroxide,
cent w/v solution of sodium nitrite, allow to stand for 2 minutes, – flow rate. 2 ml per minute,
add 2 ml of a 0.5 per cent w/v solution of ammonium – spectrophotometer set at 269 nm,
sulphamate, mix, allow to stand for 3 minutes and add 2 ml of – a 20 µl loop injector.
a 0.1 per cent w/v solution of N-(1-naphthyl)ethylenediamine The chromatographic procedure described under Assay may
dihydrochloride; a deep magenta colour is produced. be carried out. Calculate the content of C19H19N7O6 in the tablet.
526
IP 2007 FORMOTEROL FUMARATE DIHYDRATE
Assay. Determine by liquid chromatography (2.4.14). pH (2.4.24). 5.5 to 6.5, determined in a 0.1 per cent w/v solution
Test solution. Weigh and powder 20 tablets. Shake a quantity in carbon dioxide-free water.
of the powdered tablets containing about 20 mg of Folic Acid Optical rotation (2.4.22). – 0.10º to + 0.10º, determined in a 1.0
with 50 ml of 0.1 M sodium hydroxide, dilute to 100.0 ml with per cent w/v solution in methanol.
the same solvent, centrifuge and dilute 5.0 ml of the
supernatant liquid to 100.0 ml with the mobile phase. Related substances. Determine by liquid chromatography
(2.4.14).
Reference solution. Dilute 5.0 ml of a 0.02 per cent w/v solution
of folic acid RS in 0.1 M sodium hydroxide to 100.0 ml with Solvent mixture. 84 volumes of a buffer solution prepared by
the mobile phase. dissolving 6.10 g of sodium dihydrogen phosphate
monohydrate and 1.03 g of disodium hydrogen phosphate
Chromatographic system dihydrate in 1000 ml water, and 16 volumes of acetonitrile.
octadecylsilyl silica gel (10 µm), Test solution. Dissolve 20 mg of the substance under
– mobile phase: a mixture of 93 volumes of 0.05 M examination in 100 ml of the solvent mixture.
potassium dihydrogen phosphate and 7 volumes of Reference solution. A 0.00004 per cent w/v solution of
acetonitrile adjusted to pH 6.0 with 5 M sodium formoterol fumarate RS in the solvent mixture.
hydroxide,
spherical octylsilyl silica gel (5 µm)
– mobile phase: A. acetonitrile,
NOTE - For tablets containing 2 mg or less of Folic Acid the B. a buffer solution prepared by
average of the 10 individual results obtained in the test for dissolving 3.73 g of sodium dihydrogen phosphate
Uniformity of content may be taken in lieu of the Assay. monohydrate and 0.35 g of orthophosphoric acid in
Storage. Store protected from light. 1000 ml of water,
below,
Formoterol Fumarate Dihydrate – spectrophotometer set at 214 nm,
O
HN H
HO OCH3 H COOH
(min.) (per cent v/v) (per cent v/v)
CH3 , , 2H2O 0 16 84
N HOOC H
H 10 16 84
OH
2
37 70 30
(C19H24N2O4)2, C4H4O4, 2H2O Mol. Wt. 840.91 40 16 84
Formoterol Fumarate Dihydrate is dihydrate salt of fumaric 55 16 84
acid with (RS)-2′-hydroxy-5′-[(RS)-1-hydroxy-2-[[(RS)-p- Inject the reference solution. The test is not valid unless the
methoxy-α-methylphenethyl]amino]ethyl]formanilide. tailing factor is not more than 2.0 and the column efficiency in
Formoterol Fumarate Dihydrate contains not less than 98.5 not less than 4000 theoretical plates.
per cent and not more than 101.5 per cent of formoterol Inject the test solution. Any individual impurity is not more
fumarate, C42H52N4O12, calculated on the anhydrous basis. than 0.5 per cent and the sum of all the impurities is not more
Description. A white or almost white or slightly yellow powder. than 1.0 per cent.
Identification g.
Determine by infrared absorption spectrophotometry (2.4.6). Assay. Weigh accurately about 0.35 g and dissolve in 50 ml of
Compare the spectrum with that obtained with formoterol anhydrous acetic acid. Titrate with 0.1 M perchloric acid,
fumarate dihydrate RS or with the reference spectrum of determining the end-point potentiometrically (2.4.25). Carry
formoterol fumarate dihydrate. out a blank titration.
527
FORMOTEROL FUMARATE AND BUDESONIDE POWDER FOR INHALATION IP 2007
1 ml of 0.1 M perchloric acid is equivalent to 0.04024 g of Chromatographic system

C42H52N4O12. – a stainless steel column 15 cm x 4.6 mm, packed with
Storage. Store protected from light and moisture. octadecylsilyl silica gel (5 µm),
prepared by dissolving 0.138 g sodium dihydrogen
orthophosphate monohydrate and 0.122 g of decane
sulphonic acid sodium salt in 100 ml of water and
Formoterol Fumarate and Budesonide adjusting the pH to 3.0 with orthophosphoric acid, and
Powder for Inhalation 35 volumes of acetonitrile,
Formoterol Fumarate and Budesonide Powder for Inhalation – spectrophotometer set at 220 nm,
consists of Formoterol Fumarate and Budesonide in microfine – inject 200 µl.
powder either alone or admixed with Lactose in a pre-metered
Inject reference solution (c). The order of elution is formoterol
unit for use in a suitable powder inhaler.
fumarate, budesonide epimer B and epimer A. The test is not
Formoterol Fumarate and Budesonide Powder for Inhalation valid unless the column efficiency determined from the
contains not less than 90.0 per cent and not more than 125.0 formoterol fumarate and both the epimer peaks of Budesonide
per cent of the stated amounts of formoterol fumarate is not less than 1800 and 4000 theoretical plates respectively,
C42H52N4O12 and budesonide C25H34O6 per pre-metered unit. the resolution between budesonide epimer peaks is not less
than 1.5 and the relative standard deviation for formoterol
Identification fumarate and sum of peaks of budesonide epimer A and epimer
B in replicate injections is not more than 2.0 per cent.
Inject the test solution and reference solution (c).
with the test solution correspond to the peaks in the
chromatogram obtained with reference solution (c). Calculate the contents of C42H52N4O12 and C25H34O6 per unit.
Storage. Store protected from moisture, at temperature not
Tests exceeding 30º.
Other tests. Complies with the tests stated under the Inhalation Labelling. The label states the quantities of active ingredients
Preparations (Powders for Inhalation). per pre-metered unit.
Follow the procedure described under Assay with suitable
dilution of the reference solution wherever the amount of
active substance is to be determined in any test. Framycetin Sulphate
Test solution. To a suitable number of intact capsules add 10 CH2NH2
ml of water and disperse with the aid of ultrasound till the O
shells get disintegrated. Add 60 ml of the mobile phase and
mix further with the aid of ultrasound for 10 minutes with OH
intermittent shaking. Add sufficient of the mobile phase to OH
produce 100.0 ml. Dilute suitably with the mobile phase, if NH2
required, to get a final concentration of 0.6 µg per ml of O
Formoterol Fumarate in the mobile phase. H2N
OH
Reference solution (a). A 0.06 mg per ml solution of formoterol ,3H2 SO4
H2N
fumarate RS prepared by initially dissolving in 5 ml acetonitrile
and then making up to volume with the mobile phase. HOH2C O
O
Reference solution (b). A 0.2 mg per ml solution of budesonide O
RS prepared by initially dissolving in 5 ml acetonitrile and CH2NH2
then making up to volume with the mobile phase. OH O OH
HO
Reference solution (c). Dilute suitable volumes of reference NH2
solution (a) and reference solution (b) with the mobile phase
to obtain a solution containing 0.6 µg of Formoterol Fumarate
and 40 µg per ml of Budesonide per ml. C23H46N6O13,3H2SO4 Mol. Wt. 908.9
528
IP 2007 FRAMYCETIN SULPHATE
Framycetin Sulphate is 2-deoxy-4-O-(2,6-diamino-2,6- Specific optical rotation (2.4.22). +52.0° to +55.5°, determined
dideoxy-α-D-glucopyranosyl)-5-O-[3-O-(2,6-diamino-2,6- at 20° in a 10.0 per cent w/v solution.
dideoxy-β-L-idopyranosyl)-β-D-ribofuranosyl]streptamine Neamine. Determine by thin-layer chromatography (2.4.17),
(neomycin B) sulphate. The base is produced by the growth coating the plate with silica gel H.
of selected strains of Streptomyces fradiae or Streptomyces
decaris or by any other means. Mobile phase. A mixture of 30 volumes of methanol, 20 volumes
of strong ammonia solution and 10 volumes of
Framycetin Sulphate contains an amount of framycetin dichloromethane.
sulphate equivalent to not less than 630 µg of neomycin B
per mg, calculated on the dried basis. Test solution. Dissolve 0.25 g of the substance under
examination in water and dilute to 10 ml with water.
Description. A white or yellowish-white powder; odourless
or almost odourless; hygroscopic. Reference solution. Dissolve 0.5 mg of neamine RS in 2 ml of
water.
Identification Apply to the plate as 5-mm bands 5 µl of each solution. Dry
the bands and develop over a path of at least 8 cm. Dry the
plate at 105° for 10 minutes. Spray it with ninhydrin and
the plate in the following manner. Mix 0.3 g of carbomer with
stannous chloride reagent and heat at 110° for 15 minutes.
240 ml of water, allow to stand for 1 hour with moderate
Spray the plate again with the same reagent and heat at 110°
shaking, adjust the pH to 7 by the gradual addition, with
for 15 minutes. Any band corresponding to neamine in the
shaking, of 2 M sodium hydroxide and add 30 g of silica gel
H. Spread a uniform layer of the suspension 0.75 mm thick,
intense than the band in the chromatogram obtained with the
heat at 110° for 1 hour and allow to cool. Use the plate
reference solution (1 per cent).
immediately.
Neomycin C. Determine by thin-layer chromatography (2.4.17),
Mobile phase. A 10 per cent w/v solution of potassium
coating the plate with silica gel of a suitable grade.
Mobile phase. A mixture of 80 volumes of a 20 per cent w/v
solution of sodium chloride and 20 volumes of methanol.
under examination.
examination in water and dilute to 5 ml with water.
framycetin sulphate RS.
Reference solution (a). Dissolve 40 mg of framycetin sulphate
RS in water and dilute to 5 ml with water.
w/v each of framycetin sulphate RS, kanamycin sulphate RS
and streptomycin sulphate RS. Reference solution (b). Dissolve 30 mg of framycetin sulphate
RS in water and dilute to 25 ml with water. Dilute 5 ml of this
solution to 25 ml with water.
dry the plate in a current of warm air, spray with a mixture of
equal volumes of a 46 per cent w/v solution of sulphuric acid Reference solution (c). Dissolve 40 mg of neomycin sulphate
and of a 0.2 per cent w/v solution of 1,3-naphthalenediol in RS in water and dilute to 5 ml with water.
ethanol (95 per cent) and heat at 150° for about 10 minutes. Apply to the plate as 5-mm bands 10 µl of each solution. Dry
The principal spot in the chromatogram obtained with the test the bands and develop over a path of at least 12 cm. Dry the
solution corresponds to that in the chromatogram obtained plate at 100° to 105° for 10 minutes. In the chromatogram
with reference solution (a). The test is not valid unless the obtained with the test solution, the principal band corresponds
chromatogram obtained with reference solution (b) shows to the principal band in the chromatogram obtained with
three clearly separated spots. reference solution (a) and the band for neomycin C with Rf
B. Dissolve 10 mg in 5 ml of water, add 0.1 ml of pyridine and value slightly less than that of the principal band is not more
2 ml of a 0.1 per cent w/v solution of ninhydrin and heat in a intense than the band in the chromatogram obtained with
water-bath at 65° to 70° for 10 minutes; an intense violet colour reference solution (b) (3 per cent).
is produced. The test is not valid unless in the chromatogram obtained
C. A 5 per cent w/v solution gives the reactions of sulphates with reference solution (c) a band appears with Rf value slightly
(2.3.1). less than that of the principal band.
Sulphate. 27.0 to 31.0 per cent of SO4, calculated on the dried
Tests
basis, determined by the following method. Weigh accurately
pH (2.4.24). 6.0 to 7.0, determined in a 1.0 per cent w/v solution. about 0.25 g, dissolve in 100 ml of water, adjust the pH to 11
529
FRUCTOSE IP 2007
with strong ammonia solution and add 10.0 ml of 0.1 M barium free from bacterial endotoxins; (c) where applicable, that the
chloride. Titrate with 0.1 M disodium edetate using 0.5 mg of material is sterile.
metalphthalein as indicator; add 50 ml of ethanol (95 per
cent) when the colour of the solution begins to change and
continue the titration until the violet-blue colour disappears.
1 ml of 0.1 M barium chloride is equivalent to 0.009606 g of
Fructose
SO4. d-Fructose
Alcohols. Not more than 2 per cent w/w, calculated as methanol,
CH4O, determined by the following method. Dissolve 0.2 g in CH2OH
5 ml of water and add 0.05 ml of 0.05 M sulphuric acid. Distil O
the mixture and collect about 2.5 ml of the distillate in a flask. OH
OH OH
Add 25.0 ml of a 0.0167 M potassium dichromate in a mixture
OH
of 30 volumes of water and 20 volumes of sulphuric acid.
Heat on a water-bath for 30 minutes, cool and add sufficient C6H12O6 Mol. Wt. 180.2
water to produce 500.0 ml. Add 10 ml of potassium iodide
solution, allow to stand for 5 minutes and titrate with 0.1 M Fructose is D-(-)-fructopyranose.
sodium thiosulphate using starch solution, added towards Description. A white, crystalline powder with a very sweet
the end of the titration, as indicator, until the solution becomes taste.
pale green. Repeat the operation without the substance under
examination. The difference between the titrations represents Identification
the amount of potassium dichromate equivalent to the
alcohols present. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with fructose RS or
1 ml of 0.0167 M potassium dichromate is equivalent to with the reference spectrum of fructose.
0.000534 g of CH4O.
B. Dissolve 0.1 g in 10 ml of water, add 3 ml of potassium
Sulphated ash (2.3.18). Not more than 1.0 per cent. cupri-tartrate solution; the solution remains blue and clear.
Loss on drying (2.4.19). Not more than 8.0 per cent, determined Heat to boiling, a copious red precipitate is formed.
on 1.0 g by drying over phosphorus pentoxide at 60° at a C. Dissolve 5 g in water and dilute to 10 ml with the same
pressure not exceeding 0.7 kPa for 3 hours. solvent. To 0.5 ml of this solution add 0.2 g of resorcinol and
Assay. Determine by the microbiological assay of antibiotics, 9 ml of dilute hydrochloric acid and heat on a water-bath for
Method A (2.2.10), and express the results in µg of neomycin 2 minutes; a red colour is produced.
B per mg.
Tests
Framycetin Sulphate intended for administration into
internal body cavities without a further appropriate Appearance of solution. Dissolve 5.0 g in water and dilute to
procedure for removal of bacterial endotoxins complies with 10 ml with the same solvent. The solution is clear (2.4.1). Add
the following additional requirement. 10 ml of water. The solution is colourless (2.4.1).
Bacterial endotoxins (2.2.3). Not more than 1.3 Endotoxin Acidity or alkalinity. Dissolve 6.0 g in 25 ml of carbon dioxide-
Units per mg. free water and add 0.3 ml of phenolphthalein solution. The
solution is colourless. Not more than 0.15 ml of 0.1 M sodium
Framycetin Sulphate intended for administration into
hydroxide is required to change the colour of the solution to
internal body cavities without a further appropriate
pink.
additional requirement. Specific optical rotation (2.4.22). -91.0° to –93.5°, calculated
on the anhydrous basis and determined on a solution prepared
by dissolving 10.0 g in 80 ml of water, adding 0.2 ml of 5 M
Storage. Store protected from light and moisture at a ammonia, mixing well, allowing to stand for 30 minutes and
temperature not exceeding 30°. If the material is sterile, the diluting to 100.0 ml with water.
container should be tamper-evident and sealed so as to
5-Hydroxymethylfurfural and related compounds. To 5 ml of
exclude micro-organisms.
solution A add 5 ml of water and measure the absorbance
Labelling. The label states (1) the strength in terms of µg of (2.4.7) of the resulting solution at the maximum at about 284
neomycin B per mg; (b) where applicable, that the material is nm; absorbance at about 284 nm, not more than 0.32).
530
IP 2007 FRUSEMIDE
Arsenic (2.3.10). Dissolve 10.0 g in 50 ml of water and add B. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
10 ml of stannated hydrochloric acid. The resulting solution the solution remains blue and clear. Heat to boiling, a copious
complies with the limit test for arsenic (1 ppm). red precipitate is formed.
Heavy metals (2.3.13). A solution of 4.0 g in 10 ml of water, 2 ml Tests
of dilute acetic acid and sufficient water to produce 25.0 ml
complies with the limit test for heavy metals, Method A pH (2.4.24). 3.0 to 6.0, determined in a solution diluted, if
(5 ppm). necessary, with water for injections to contain not more than
5.0 per cent w/v of Fructose and to which 0.30 ml of a saturated
Chlorides (2.3.12). 20 ml of a 10 per cent w/v solution (solution solution of potassium chloride has been added for each
A) complies with the limit test for chlorides (125 ppm). 100 ml of solution.
Sulphates (2.3.17). 7.5 ml of solution A diluted to 15 ml with 5-Hydroxymethylfurfural and Related substances. Dilute a
water complies with the limit test for sulphates (200 ppm). volume containing 1.0 g of Fructose to 500.0 ml with water
Foreign sugars. Dissolve 5.0 g in water and dilute to 10 ml and measure the absorbance (2.4.7) of the resulting solution
with the same solvent. To 1 ml of the solution add 9 ml of at the maximum at about 284 nm; absorbance at about 284 nm,
ethanol (95 per cent). Any opalescence in the solution is not not more than 0.50.
more intense than that in a mixture of 1 ml of the initial solution Heavy metals (2.3.13). A volume of the injection containing
and 9 ml of water. 4.0 g of fructose that has been evaporated to a volume of
Barium. To 10 ml of solution A add 1 ml of 1 M sulphuric acid. about 10 ml, cooled and diluted to 25 ml with water complies
Examine exactly after 1 hour; any opalescence in the solution with the limit test fpr heavy metals, Method A (5 ppm).
is not more intense than that in a mixture of 10 ml of solution A Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit
and 1 ml of water. per ml.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Other tests. Complies with the tests stated under Parenteral
Water (2.3.43). Not more than 0.5 per cent, determined on Preparations (Injections).
1.0 g. Assay. To an accurately measured volume containing about
5.0 g of Fructose, add 0.2 ml of 5 M ammonia and sufficient
Fructose intended for use in the manufacture of parenteral
water to produce 100.0 ml. Mix well, allow to stand for
preparations without a further appropriate procedure for
30 minutes and determine the optical rotation in a 2-dm tube
the removal of bacterial endotoxins complies with the
(2.4.22). The observed rotation in degrees multiplied by 0.5427
represents the weight, in g, of fructose, C6H12O6, in the volume
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit taken for Assay.
per ml of a 5 per cent w/v solution in water for injections.
Storage. Store in single dose containers at a temperature not
Storage. Store protected from moisture. exceeding 30°.
Labelling. The label states whether or not the contents are Labelling. The label states (1) the strength as the percentage
intended for use in the manufacture of parenteral preparations. w/v of fructose, C6H12O6; (2) that the injection should not be
used if it contains visible particles.
Fructose Injection
Frusemide
Fructose Intravenous Infusion
Furosemide
Fructose Injection is a sterile solution of Fructose in Water for
Injections. OH
O
Fructose Injection contains not less than 95.0 per cent and H
not more than 105.0 per cent of the stated amount of fructose, N
O
C6H12O6. It contains no antimicrobial agent.
H2N
Description. A clear, colourless solution. S
O O Cl
Identification
A. The solution prepared as directed in the Assay is laevo- C12H11ClN2O5S Mol. Wt. 330.7
rotatory. Frusemide is 4-chloro-N-furfuryl-5-sulphamoylanthranilic acid.
531
FRUSEMIDE INJECTION IP 2007
Frusemide contains not less than 98.5 per cent and not more Sulphated ash (2.3.18). Not more than 0.1 per cent.
than 101.0 per cent of C12H11ClN2O5S, calculated on the dried Loss on drying (2.4.19). Not more than 0.5 per cent, determined
basis. on 1.0 g by drying in an oven at 105°.
Description. A white or almost white, crystalline powder. Assay. Weigh accurately about 0.5 g, dissolve in 40 ml of
dimethylformamide and titrate with 0.1 M sodium hydroxide
Identification using bromothymol blue solution as indicator. Carry out a
Test A may be omitted if tests B and C are carried out. Tests B blank titration.
and C may be omitted if test A is carried out. 1 ml of 0.1 M sodium hydroxide is equivalent to 0.03307 g of
A. Determine by infrared absorption spectrophotometry (2.4.6). C12H11ClN2O5S.
Compare the spectrum with that obtained with frusemide RS Storage. Store protected from light.
or with the reference spectrum of frusemide.
0.0005 per cent w/v solution in 0.1 M sodium hydroxide shows Frusemide Injection
three absorption maxima at about 228 nm, 271 nm and 333 nm.
Furosemide Injection
The ratio of the absorbance at the maximum at about 271 nm to
that at the maximum at about 228 nm is 0.52 to 0.57. Frusemide Injection is a sterile solution of Frusemide in Water
for Injections prepared with the aid of Sodium Hydroxide.
C. Dissolve about 5 mg in 10 ml of methanol. Transfer 1 ml of
this solution to a flask, add 10 ml of dilute hydrochloric acid Frusemide Injection contains not less than 95.0 per cent and
and boil under a reflux condenser on a water-bath for not more than 105.0 per cent of the stated amount of frusemide,
15 minutes. Cool, add 15 ml of 1 M sodium hydroxide and 5 ml C12H11ClN2O5S.
of a 0.1 per cent w/v solution of sodium nitrite. Allow to stand Description. A clear, colourless or almost colourless solution.
for 3 minutes, add 5 ml of a 0.5 per cent w/v solution of
ammonium sulphamate, mix and add 5 ml of a 0.1 per cent Identification
w/v solution of N-(1-naphthyl)ethylenediamine
dihydrochloride; a red-violet colour is produced. A. When examined in the range 220 nm to 360 nm (2.4.7), the
final solution obtained in the Assay shows three absorption
Tests maxima at about 228 nm, 271 nm and 333 nm.
Heavy metals (2.3.13). 1.0 g complies with the limit test for B. To a volume of the injection containing 5 mg of Frusemide
heavy metals, Method B (20 ppm). add 10 ml of dilute hydrochloric acid and boil under a reflux
condenser on a water-bath for 15 minutes. Cool, add 15 ml of
Chlorides (2.3.12). Shake 1 g with 40 ml of water for 5 minutes 1 M sodium hydroxide and 5 ml of a 0.1 per cent w/v solution
and filter. The filtrate complies with the limit test for chlorides of sodium nitrite. Allow to stand for 3 minutes, add 5 ml of a
(250 ppm). 0.5 per cent w/v solution of ammonium sulphamate, mix and
Sulphates (2.3.17). Shake 1.0 g with a mixture of 30 ml of add 5 ml of a 0.1 per cent w/v solution of N- (1-naphthyl)
distilled water and 0.2 ml of 5 M acetic acid for 5 minutes and ethylenediamine dihydrochloride; a red-violet colour is
filter. 15 ml of the filtrate complies with the limit test for produced.
sulphates (300 ppm). Tests
Free amines. Dissolve 0.1 g in 25 ml of methanol. To 1 ml of
pH (2.4.24). 8.0 to 9.3.
the solution add 3 ml of dimethylformamide, 12 ml of water
and 1 ml of 1 M hydrochloric acid. Cool, add 1 ml of a 0.5 per Free amines. To a volume of the injection containing 0.1 g of
cent w/v solution of sodium nitrite, shake and allow to stand Frusemide add sufficient methanol to produce 25 ml, shake
for 5 minutes. Add 1 ml of a 2.5 per cent w/v solution of and filter. To 1 ml of the filtrate add 3 ml of dimethylformamide,
sulphamic acid, shake and allow to stand for 3 minutes. Add 12 ml of water and 1 ml of 1 M hydrochloric acid. Cool, add 1
1 ml of a 0.5 per cent w/v solution of N- (1-naphthyl)- ml of a 0.5 per cent w/v solution of sodium nitrite, shake and
ethylenediamine dihydrochloride and dilute to 25 ml with allow to stand for 5 minutes. Add 1 ml of a 2.5 per cent w/v
water. Measure the absorbance of the resulting solution at solution of sulphamic acid, shake and allow to stand for
the maximum at about 530 nm (2.4.7), using as the blank a 3 minutes. Add 1 ml of a 0.5 per cent w/v solution of N-(1-
solution obtained by treating 1 ml of methanol and 3 ml of naphthyl)ethylenediamine dihydrochloride and dilute to 25
dimethylformamide in a similar manner; absorbance is not ml with water. Measure the absorbance of the resulting solution
more than 0.12. at the maximum at about 530 nm (2.4.7), using as the blank a
532
IP 2007 FURAZOLIDONE
solution obtained by treating 1 ml of methanol and 3 ml of Other tests. Comply with the tests stated under Tablets.
dimethylformamide in a similar manner; absorbance is not Assay. Weigh and powder 20 tablets. Weigh accurately a
more than 0.20. quantity of the powder containing about 0.1 g of Frusemide
Bacterial endotoxins (2.2.3). Not more than 3.5 Endotoxin Units and shake with 150 ml of 0.1 M sodium hydroxide for 10
per mg of frusemide. minutes. Add sufficient 0.1 M sodium hydroxide to produce
Other tests. Complies with the tests stated under Parenteral 250.0 ml and filter. Dilute 5.0 ml to 200.0 ml with 0.1 M sodium
Preparations (Injections). hydroxide and measure the absorbance of the resulting
Assay. Dilute an accurately measured volume of the injection content of C12H11ClN2O5S taking 580 as the specific absorbance
containing about 20 mg of Frusemide with water to produce at 271 nm.
100.0 ml. Dilute 5.0 ml to 100.0 ml with 0.1 M sodium hydroxide.
Measure the absorbance of the resulting solution at the Storage. Store protected from light.
C12H11ClN2O5S taking 580 as the specific absorbance at
271 nm.
Furazolidone
H
O C O
O2N N
Frusemide Tablets N
O
Furosemide Tablets
Frusemide Tablets contain not less than 90.0 per cent and not C8H7N3O5 Mol. Wt. 225.2
more than 110.0 per cent of the stated amount of frusemide, Furazolidone is 3-(5-nitrofurfurylideneamino)oxazolidin-2-
C12H11ClN2O5S. one
Identification Furazolidone contains not less than 97.0 per cent and not
more than 103.0 per cent of C8H7N3O5, calculated on the dried
basis.
final solution obtained in the Assay shows three absorption
maxima at about 228 nm, 271 nm and 333 nm. Description. A yellow, crystalline powder; odourless or almost
odourless.
B. Shake a quantity of the powdered tablets containing 80 mg
of Frusemide with 10 ml of ethanol (95 per cent), filter and
Identification
evaporate the filtrate to dryness. Dissolve 25 mg of the residue
obtained in 2.5 ml of ethanol (95 per cent) and add 2 ml of 4- A. Determine by infrared absorption spectrophotometry (2.4.6).
dimethylaminobenzaldehyde solution; a green colour is Compare the spectrum with that obtained with furazolidone
produced which changes to deep red. RS or with the reference spectrum of furazolidone.
Tests B. Dissolve 1 mg in 1 ml of dimethylformamide and add 0.05 ml

of 1 M ethanolic potassium hydroxide; a deep blue colour is
Free amines. To a quantity of the powdered tablets containing produced.
0.1 g of Frusemide add 25 ml of methanol, shake and filter. To
1 ml of the filtrate add 3 ml of dimethylformamide, 12 ml of Tests
water and 1 ml of 1 M hydrochloric acid. Cool, add 1 ml of a
0.5 per cent w/v solution of sodium nitrite, shake and allow to pH (2.4.24). 4.5 to 7.0, determined in a solution prepared by
stand for 5 minutes. Add 1 ml of a 2.5 per cent w/v solution of shaking 1.0 g for 15 minutes with 100 ml of carbon dioxide-
sulphamic acid, shake and allow to stand for 3 minutes. Add free water and filtering.
1 ml of a 0.5 per cent w/v solution of N-(1-naphthyl) Nitrofurfural diacetate. Determine by thin-layer
ethylenediamine dihydrochloride and dilute to 25 ml with chromatography (2.4.17), coating the plate with silica gel G.
water. Measure the absorbance of the resulting solution at
the maximum at about 530 nm (2.4.7), using as the blank a
volumes of dioxan.
solution obtained by treating 1 ml of methanol and 3 ml of
dimethylformamide in a similar manner; the absorbance is not Test solution. Dissolve 50 mg of the substance under
more than 0.20. examination in 5 ml of dimethylformamide by heating on a
533
FURAZOLIDONE ORAL SUSPENSION IP 2007
water-bath for a few minutes, allow to cool and dilute to 10 ml Other tests. Complies with the tests stated under Oral Liquids.
with acetone. Assay. Carry out the following procedure protected from
Reference solution. A solution containing 0.01 per cent w/v of light.
nitrofurfural diacetate RS in a mixture of equal volumes of
To an accurately measured volume of the suspension
dimethylformamide and acetone.
containing about 50 mg of Furazolidone add 5 ml of water and
Apply to the plate 20 µl of the test solution and 10 µl of the mix. Transfer this mixture to a 250-ml volumetric flask with the
reference solution. After development, dry the plate in air and aid of dimethylformamide. Add about 150 ml of
heat it at 105° for 5 minutes. Spray with a solution prepared by dimethylformamide, shake by mechanical means for 10
dissolving 0.75 g of phenylhydrazine hydrochloride in 10 ml minutes, dilute to volume with dimethylformamide and mix.
of ethanol (95 per cent), diluting to 50 ml with water, adding Dilute 5.0 ml of this solution to 100.0 ml with water and mix
activated charcoal, filtering and then adding 25 ml of well. Measure the absorbance of the resulting solution at the
hydrochloric acid and sufficient water to produce 200 ml. maximum at about 367 nm (2.4.7), using dimethylformamide
Any spot corresponding to nitrofurfural diacetate in the solution (5 per cent v/v) as the blank. Calculate the content of
chromatogram obtained with the test solution is not more C8H7N3O5 taking 750 as the specific absorbance at 367 nm.
reference solution.
exceeding 30°.
Furazolidone Tablets
Assay. Carry out the following procedure protected from
light. Furazolidone Tablets contain not less than 90.0 per cent and
Weigh accurately about 80 mg, add 150 ml of furazolidone, C8H7N3O5.
dimethylformamide, swirl to dissolve and add sufficient water
to produce 500.0 ml. Dilute 5.0 ml to 100.0 ml with water and Identification
mix. Measure the absorbance of the resulting solution at the
maximum at about 367 nm (2.4.7). Calculate the content of To a quantity of the powdered tablets containing 50 mg of
C8H7N3O5 taking 750 as the specific absorbance at 367 nm. Furazolidone add 10 ml of a freshly prepared mixture of
9 volumes of dimethylformamide and 1 volume of 1 M
Storage. Store protected from light. ethanolic potassium hydroxide. The solution turns purple,
immediately changes to deep blue and on standing for 10
minutes, again turns purple.
Furazolidone Oral Suspension Tests

Furazolidone Oral Suspension is a suspension of Furazolidone Other tests. Comply with the tests stated under Tablets.
in a suitable aqueous flavoured vehicle.
Assay. Carry out the following procedure protected from
Furazolidone Oral Suspension contains not less than 90.0 per light.
furazolidone, C8H7N3O5.
the powder containing about 80 mg of Furazolidone into a
Identification 200-ml volumetric flask, add 100 ml of dimethylformamide,
warm to about 50° and shake well. Cool, dilute to volume with
Add a quantity of the suspension containing 50 mg of dimethylformamide, mix and centrifuge a small quantity of the
Furazolidone to 10 ml of a freshly prepared mixture of 9 volumes mixture. Dilute 5.0 ml of the clear, supernatant liquid to 250.0
of dimethylformamide and 1 volume of 1 M ethanolic ml with water and mix. Measure the absorbance of the resulting
potassium hydroxide. The solution turns purple, immediately solution at the maximum at about 367 nm (2.4.7), using
changes to deep blue and on standing for about 10 minutes, dimethylformamide diluted 50 times with water as the blank.
again turns purple. Calculate the content of C8H7N3O5 taking 750 as the specific
Tests
pH (2.4.24). 6.0 to 8.5. exceeding 30°.
534
IP 2007 FUSIDIC ACID ORAL SUSPENSION
Fusidic Acid Reference solution (a). Dissolve 5 mg of 3-ketofusidic acid

RS in 5 ml of the mobile phase. To 1 ml of this solution add 0.2
ml of the test solution and dilute to 20 ml with the mobile
H phase.
H3C
COOH Reference solution (b). Dilute 2.0 ml of the test solution to 10
H3C H ml with the mobile phase.
HO
1 Chromatographic system
H3C H3C OCOCH3 , /2H2O
– a stainless steel column 12.5 cm x 4 mm, packed with
H CH3 octadecylsilyl silica gel (5 µm),
HO – mobile phase: a mixture of 50 volumes of acetonitrile,
H 20 volumes of water, 20 volumes of a 1 per cent w/v
CH3
solution of phosphoric acid and 10 volumes of
methanol,
C31H48O6,½H2O Mol. Wt. 525.7 – flow rate. 2 ml per minute,
Fusidic Acid is ent-16α-acetoxy-3β,11β-dihydroxy- – spectrophotometer set at 235 nm,
4β,8β,14β-trimethyl-18-nor-5β,10α-cholesta(17Z)-17(20),24- – a 20 µl loop injector.
dien-21-oic acid hemihydrate, an antimicrobial substance Continue the chromatography for at least 3.5 times the retention
produced by the growth of certain strains of Fusidium time of the principal peak. In the chromatogram obtained with
coccineum or by any other means. the test solution the sum of the areas of any secondary peaks
Fusidic Acid contains not less than 97.5 per cent and not more is not greater than 4 times the area of the peak corresponding
than 101.0 per cent of C31H48O6, calculated on the anhydrous to fusidic acid in the chromatogram obtained with reference
basis. solution (a). Ignore any peak with an area less than that of the
solution (b). The test is not valid unless the resolution factor
Identification between the peaks corresponding to 3-ketofusidic acid and
fusidic acid in the chromatogram obtained with reference
A. Determine by infrared absorption spectrophotometry (2.4.6). solution (a) is not less than 2.5 and unless the principal peak
Compare the spectrum with that obtained with fusidic acid RS in the chromatogram obtained with reference solution (b) has
or with the reference spectrum of fusidic acid. a signal-to-noise ratio of not less than 3.
B. Determine by thin-layer chromatography (2.4.17), coating Sulphated ash (2.3.18). Not more than 0.2 per cent.
the plate with silica gel G. Water (2.3.43). 1.4 to 2.0 per cent, determined on 0.5 g.
Mobile phase. A mixture of 80 volumes of chloroform, 10 Assay. Weigh accurately about 0.5 g and dissolve in 10 ml of
volumes of glacial acetic acid, 10 volumes of cyclohexane ethanol (95 per cent). Titrate with 0.1 M sodium hydroxide
and 2.5 volumes of methanol. using phenolphthalein solution as indicator.
Test solution. Dissolve 0.2 g of the substance under 1 ml of 0.1 M sodium hydroxide is equivalent to 0.05167 g of
examination in 100 ml of ethanol (95 per cent). C31H48O6.
Reference solution. A 0.24 per cent w/v solution of Storage. Store protected from light.
diethanolamine fusidate RS in ethanol (95 per cent).
dry the plate at 105° for 10 minutes and examine in ultraviolet
light at 365 nm. The principal spot in the chromatogram Fusidic Acid Oral Suspension
chromatogram obtained with the reference solution. Fusidic Acid Mixture
Fusidic Acid Oral Suspension is a suspension of Fusidic Acid
Tests
in powder of suitable fineness in a suitable flavoured vehicle.
Related substances. Determine by liquid chromatography It should not be diluted.
(2.4.17). Fusidic Acid Oral Suspension contains not less than 92.5 per
Test solution. Dissolve 0.5 g of the substance under cent and not more than 107.5 per cent of the stated amount of
examination in 100 ml of the mobile phase. anhydrous fusidic acid, C31H48O6.
535
FUSIDIC ACID ORAL SUSPENSION IP 2007
Identification and examine in ultraviolet light at 365 nm. Any secondary spot
in the chromatogram obtained with the test solution is not
A. To a quantity of the oral suspension containing 0.1 g of more intense than the principal spot in the chromatogram
anhydrous fusidic acid add 5 ml of water and extract with obtained with reference solution (a). Any yellow spot in the
three quantities, each of 10 ml, of chloroform. Wash the chromatogram obtained with the test solution is not more
combined extracts with two quantities, each of 10 ml, of water, intense than the principal spot in the chromatogram obtained
dry with anhydrous sodium sulphate, evaporate to dryness with reference solution (b).
and dissolve the residue in 1 ml of chloroform IR. The resulting
solution complies with the following test. Other tests. Complies with the tests stated under Oral Liquids.
Determine by infrared absorption spectrophotometry (2.4.6). Assay. Determine by liquid chromatography (2.4.14).
Compare the spectrum obtained with that obtained with fusidic Test solution. Shake thoroughly an accurately weighed
acid RS or with the reference spectrum of fusidic acid. quantity of the oral suspension containing about 30 mg of
B. In the Assay, the principal peak in the chromatogram anhydrous fusidic acid with 25 ml of the mobile phase, dilute
obtained with the test solution corresponds to the peak due to 50.0 ml with the mobile phase, filter and use the clear filtrate.
to fusidic acid in the chromatogram obtained with the reference Reference solution. A solution containing 0.07 per cent w/v of
solution. diethanolamine fusidate RS in the mobile phase.
Tests Chromatographic system

pH (2.4.24). 4.8 to 5.2. octadecylsilyl silica gel (5 µm) (such as LiChrosorb RP-
Related substances. Determine by thin-layer chromatography 18),
(2.4.17), coating the plate with silica gel G. – mobile phase: a mixture of 60 volumes of acetonitrile,
30 volumes of a 1 per cent v/v solution of glacial acetic
Mobile phase. A mixture of 160 volumes of chloroform, 40 acid and 10 volumes of methanol,
volumes of cyclohexane, 40 volumes of glacial acetic acid – flow rate. 1.5 ml per minute,
and 5 volumes of methanol. – spectrophotometer set at 235 nm,
Test solution. Extract a quantity of the oral suspension – a 20 µl loop injector.
containing 0.1 g of anhydrous fusidic acid with two quantities, The assay is not valid unless the column efficiency,
each of 20 ml, of chloroform, evaporate the combined extracts determined using the principal peak in the chromatogram
to dryness and dissolve the residue in 10 ml of ethanol (95 obtained with the reference solution, is not less than 14,000
per cent). theoretical plates per metre.
Reference solution (a). A solution containing 0.04 per cent w/ Determine the weight per ml of the oral suspension (2.4.29),
v of diethanolamine fusidate RS in ethanol (95 per cent). and calculate the content of C31H48O6, weight in volume.
Reference solution (b). A solution containing 0.04 per cent w/ Storage. Store protected from light at a temperature not
v of 3-ketofusidic acid RS in ethanol (95 per cent). exceeding 30°.
Apply to the plate 5 µl of each solution. After development, Labelling. The label states the quantity of active ingredient in
dry the plate at 110° for 10 minutes, spray with ethanolic terms of the equivalent amount of anhydrous fusidic acid.
sulphuric acid (10 per cent v/v), dry at 110° for 10 minutes
536
G
Gallamine Triethiodide ....
Gallamine Injection ....
Gatifloxacin ....
Gatifloxacin Infusion ....
Gatifloxacin Tablets ....
Gelatin ....
Hard Gelatin Capsule Shells ....
Gentamicin Sulphate ....
Gentamicin Eye Drops ....
Gentamicin Injection ....
Glibenclamide ....
Glibenclamide Tablets ....
Glipizide ....
Glipizide Tablets ....
2-Deoxy-D-Glucose ....
Glycerin ....
Glyceryl Monostearate ....
Concentrated Glyceryl Trinitrate Solution ....
Glyceryl Trinitrate Tablets ....
Glycine ....
Glycine Irrigation Solution ....
Griseofulvin ....
Griseofulvin Tablets ....
Guaiphenesin ....
537
IP 2007 GALLAMINE INJECTION
Gallamine Triethiodide Test solution. Dissolve 0.5 g of the substance under

examination in 100 ml of ethanol (95 per cent).
N(C2H5)3 Reference solution. Dilute 1 volume of the test solution to 100

O volumes with ethanol (95 per cent).
O O Apply to the plate 10 µl of each solution. After development,
(C2H5)3 N N(C2H5 )3 3I
dry the plate in a current of warm air and spray with
iodoplatinate reagent. In the chromatogram obtained with
the test solution an elongated blue spot, which may appear to
C30H60I3N3O3 Mol. Wt. 891.5 be double, is produced. Any secondary spot in the
Gallamine Triethiodide is 2,2',2''-(benzene-1,2,3- chromatogram obtained with the test solution of higher Rf
triyltrioxy)tris(tetraethylammonium) triiodide. value than the principal spot is not more intense than the
principal spot in the chromatogram obtained with the reference
Gallamine Triethiodide contains not less than 98.0 per cent
solution.
and not more than 101.0 per cent of C30H60I3N3O3, calculated
on the dried basis. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Description. A white or almost white powder; odourless or Loss on drying (2.4.19). Not more than 1.5 per cent, determined
almost odourless; hygroscopic. on 1.0 g by drying in an oven at 105°.
Identification of 40 ml of acetone and 15 ml of mercuric acetate solution.
Test A may be omitted if tests B, C and D are carried out. Tests Titrate with 0.1 M perchloric acid, determining the end-point
B and C may be omitted if tests A and D are carried out. potentiometrically (2.4.25). Carry out a blank titration.
Compare the spectrum with that obtained with gallamine C30H60I3N3O3.
triethiodide RS. Storage. Store protected from light.
an absorption maximum at about 225 nm; absorbance at about Gallamine Injection
225 nm, 0.50 to 0.55.
Gallamine Triethiodide Injection
C. To 5 ml of a 1 per cent w/v solution add 1 ml of potassium
mercuri-iodide solution; a yellow precipitate is produced. Gallamine Injection is a sterile solution of Gallamine
Triethiodide in Water for Injections.
D. Acidify 2 ml of a 0.5 per cent w/v solution with 0.2 ml of 2 M
Gallamine Injection contains not less than 95.0 per cent and
nitric acid; the resulting solution gives reaction A of iodides
not more than 105.0 per cent of the stated amount of gallamine
(2.3.1).
triethiodide, C30H60I3N3O3.
Tests Description. A clear, colourless or almost colourless solution.
Appearance of solution. A 2.0 per cent w/v solution is clear Identification
(2.4.1), and, when examined immediately after preparation, not
more intensely coloured than reference solution YS7 (2.4.1). A. When examined in the range 220 nm to 360 nm (2.4.7), the
final solution obtained in the Assay shows an absorption
Acidity or alkalinity. To 50 ml of water add 0.2 ml of methyl
maximum only at about 225 nm.
red solution and adjust the pH to 6 by adding either 0.01 M
sulphuric acid or 0.02 M sodium hydroxide until the colour is B. To 1 ml add 1 ml of iodinated potassium iodide solution; a
orange-yellow. Add 1 g of the substance under examination brown precipitate is produced.
and shake to dissolve. Not more than 0.2 ml of 0.01 M C. To 1 ml add 1 ml of potassium mercuri-iodide solution; a
sulphuric acid or 0.02 M sodium hydroxide is required to yellow precipitate is produced.
restore the orange-yellow colour.
Tests
(2.4.17), coating the plate with microcrystalline cellulose. pH (2.4.24). 5.5 to 7.5.
Mobile phase. A mixture of 66 volumes of 1-butanol, 17 Other tests. Complies with the tests stated under Parenteral
volumes of glacial acetic acid and 17 volumes of water. Preparations (Injections).
539
GATIFLOXACIN IP 2007
Assay. Dilute an accurately measured volume containing about – mobile phase: a mixture of 85 volumes of buffer solution
40 mg of Gallamine Triethiodide with sufficient 0.01 M prepared by dissolving 4.1 g of sodium acetate
hydrochloric acid to produce 200.0 ml. Dilute 5.0 ml of this anhydrous in 1000 ml with water, add 4 ml triehylamine.
solution to 100.0 ml with 0.01 M hydrochloric acid and measure Adjust the pH to 4.0 with acetic acid and 15 volumes of
the absorbance of the resulting solution at the maximum at acetonitrile,
about 225 nm (2.4.7). Calculate the content of C30H60I3N3O3 – flow rate. 1 ml per minute,
taking 525 as the specific absorbance at 225 nm. – spectrophotometer set at 295 nm,
Storage. Store protected from light. – a 20 µl loop injector.
Gatifloxacin
CH3 secondary peak is not more than 0.5 times the area of the peak
HN H3CO
N N not more than the area of the peak in the chromatogram
, 11/2 H2O obtained with the reference solution (b) (1.0 per cent).
F COOH Heavy metals (2.3.13). 1.0 g complies with the limit test for
O heavy metals, Method B (20 ppm).
C19H22FN3O4 Wt. 402.4
Gatifloxacin is (RS)-1-cyclopropyl-6-fluoro-1,4-dihydro-8-
methoxy-7-(3-methylpiperazin-1-yl)-4-oxo-3- Assay. Weigh accurately about 0.3 g, dissolve in 20 ml of
quinolinecarboxylic acid sesquihydrate N,N’-dimethylformamide, add 50 ml of methanol. Titrate with
0.1 M hydrochloric acid. Determine the end-point
Gatifloxacin contains not less than 98.0 per cent and not more
than 102.0 per cent of C19H22FN3O4, calculated on the anhydrous
basis. 1 ml 0.1 M hydrochloric acid is equivalent of 0.03754 g of
C19H22FN3O4,1.5H2O.
Description. A white to light yellow crystalline powder.
Identification
Compare the spectrum with that obtained with gatifloxacin
RS. Gatifloxacin Infusion
Tests Gatifloxacin Infusion contains Gatifloxacin.
Gatifloxacin Infusion contain not less than 90.0 per cent and
(2.4.14).
gatifloxacin C19H22FN3O4.
examination in 100 ml of the mobile phase. Identification
Reference solution (a). A 0.03 per cent w/v solution of In the Assay, the principal peak in the chromatogram obtained
gatifloxacin RS in the mobile phase. with the test solution corresponds to the peak in the
Reference solution (b). Dilute 1 ml of reference solution (a) to chromatogram obtained with the reference solution.
Tests
– a stainless steel column 25 cm x 4.6 mm packed with pH (2.4.24). 3.5 to 5.5.
octadecylsilane bonded to porous silica (5µm), Related substances. Determine by liquid chromatography
– column temperature 50º, (2.4.14).
540
IP 2007 GATIFLOXACIN TABLETS
NOTE — Protect the solutions from light. Inject the test solution and the reference solution.
Test solution. Dilute the Infusion to obtain 0.1 per cent w/v Calculate the content of C19H22FN3O4.
solution in mobile phase. Storage. Store protected from light and moisture.
gatifloxacin RS in the mobile phase.
Reference solution (b). Dilute 1 ml of reference solution (a) to Gatifloxacin Tablets
Gatifloxacin Tablets contain not less than 90.0 per cent and
Chromatographic system not more than 110.0 per cent of the stated amount of
– a stainless steel column 25 cm x 4.6 mm packed with gatifloxacin, C19H22FN3O4.
– mobile phase: a mixture of 80 volumes of buffer solution Identification
prepared by dissolving 1.6 ml of orthophosphoric acid
in 1000 ml of water. Adjust the pH 3.0 with triethylamine, In the Assay, the principal peak in the chromatogram obtained
– flow rate. 1 ml per minute, with the test solution corresponds to the peak in the
– spectrophotometer set at 240 nm, chromatogram obtained with the reference solution.
Tests
effciency is not less than 2000 theoretical plates and the tailing Dissolution (2.5.2).
factor is not more than 2.0. Apparatus. No 1
Inject the test solution and reference solution (b). In the Medium. 900 ml of acetate buffer pH 4.0.
chromatogram obtained with the test solution, the area of any Speed and time. 50 rpm for 45 minutes.
secondary peak is not more than the area of the peak in the Withdraw a suitable volume of the medium and filter.
cent) and the sum of areas of all the secondary peaks is not Determine by liquid chromatography (2.4.14).
more than twice the area of the peak in the chromatogram Test solution. The filtrate obtained as given above.
Reference solution. A 0.1 per cent w/v solution of gatifloxacin
Other tests. Comply with the tests stated under Infusion. RS in dissolution medium.
Assay. Determine by liquid chromatography (2.4.14). Chromatographic system
NOTE — Protect the solutions from light.
Test solution. Dilute the Infusion to obtain 0.1 per cent w/v – mobile phase: a mixture of 80 volumes of 1.6 per cent
solution in mobile phase. v/v solution of phosphoric acid adjusted to pH 3.0
Reference solution. A 0.1 per cent w/v solution of gatifloxacin with triethylamine and 20 volumes of acetonitrile,
RS in the mobile phase. – flow rate. 1 ml per minute,
Chromatographic system – a 50 µl loop injector.
C19H22FN3O4.
– mobile phase: a mixture of 80 volumes of buffer solution
prepared by dissolving 1.6 ml of orthophosphoric acid Related substances. Determine by liquid chromatography
in 1000 ml of water, adjust to pH 3.0 with triethylamine, (2.4.14).
– flow rate. 1 ml per minute, NOTE — Protect the solutions from light.
– a 20 µl loop injector. Test solution. Dilute the Infusion to obtain 0.1 per cent w/v
solution in mobile phase.
column efficiency determined from the principal peak is not Reference solution (a). A 0.1 per cent w/v solution of
less than 2500 theoretical plates, the tailing factor is not more gatifloxacin RS in the mobile phase.
than 3.0 and the relative standard deviation of five replicate Reference solution (b). Dilute 1 ml of reference solution (a) to
injections is not more than 2.0 per cent. 100 ml with mobile phase.
541
GELATIN IP 2007
Chromatographic system as described under Assay. Identification

Inject reference solution (a). Test is not valid unless the column A. Dissolve 1 g in sufficient carbon dioxide-free water at
effciency is not less than 2000 theoretical plates and the tailing about 55° to produce 100 ml and maintain the solution at this
factor is not more than 2.0. temperature until required for use (solution A). To 2 ml of
Inject the test solution and reference solution (b). In the solution A add 0.05 ml of copper sulphate solution, mix and
chromatogram obtained with the test solution, the area of any add 0.5 ml of 2 M sodium hydroxide; a violet colour is produced.
B. Add 10 ml of water to 0.5 g in a test-tube, allow to stand for
10 minutes, heat at 60° for 15 minutes, allow to stand upright
at 0° for 6 hours and invert the test-tube; the contents do not
more than 3 times the area of the peak in the chromatogram
immediately flow out.
Assay. Determine by liquid chromatography (2.4.14). Appearance of solution. Solution A is not more opalescent
Test solution. Weigh and powder 20 tablets. Weigh accurately than opalescence standard OS4 (2.4.1), and not more intensely
a quantity of the powder containing 100 mg of Gatifloxacin, coloured than reference solution YS4 (2.4.1).
dissolve in 50 ml of 0.1 per cent phosphoric acid solution pH (2.4.24). 3.8 to 7.6, determined in solution A.
and dilute to 100 ml with same solvent and filter. Dilute 5 ml of
the solution to 10 ml with mobile phase. Arsenic (2.3.10). To 5.0 g add 10 ml of water and allow to
stand for 1 hour. Warm to dissolve and add 10 ml of
Reference solution. A 0.05 per cent of gatifloxacin RS in mobile
hydrochloric acid and a slight excess of bromine solution.
phase.
Add 2 ml of stannated hydrochloric acid, heat under a reflux
Chromatographic system condenser for 1 hour, cool and add 10 ml of water and 10 ml of
– a stainless steel column 25 cm x 4.6 mm packed with hydrochloric acid. The resulting solution complies with the
octadecylsilane bonded to porous silica (5 µm), limit test for arsenic (2 ppm).
– mobile phase: a mixture of 80 volumes of 1.6 per cent
Heavy metals (2.3.13). To the residue obtained in the test for
v/v solution of phosphoric acid adjusted to pH 3.0
Ash add 2 ml of hydrochloric acid and 0.5 ml of nitric acid,
with triethylamine and 20 volumes of acetonitrile,
and evaporate to dryness. To the residue add 1 ml of 1 M
hydrochloric acid and 15 ml of water and warm for a few
minutes. Filter, wash with water and dilute the filtrate to 100 ml
with water. Dilute 8 ml of this solution to 25 ml with water. The
Inject the reference solution. The test is not valid unless the resulting solution complies with the limit test for heavy metals,
tailing factor is not more than 3.0. The column efficiency in Method A (50 ppm).
not less than 2500 theoretical plates. The relative standard
deviation for replicate injections is not more than 2.0 per cent. Sulphur dioxide. Not more than 200 ppm, determined by the
following method. Add 150 ml of water to a 500-ml three-
Inject the test solution and the reference solution. necked, round-bottomed flask fitted with a water-cooled reflux
Calculate the content of C19H22FN3O4. condenser 200 mm long the upper end of which is connected
to an absorption tube. The flask is fitted with a 100-ml dropping
funnel and a gas inlet tube that reaches nearly to the bottom
of the flask. Pass a stream of carbon dioxide through the flask
at a rate of 100 ml per minute for 15 minutes. Connect an
Gelatin absorption tube containing 10 ml of hydrogen peroxide
solution (10 vol) previously neutralised to a 0.1 per cent w/v
Gelatin is a purified protein obtained by partial hydrolysis of solution of bromophenol blue in ethanol (20 per cent) and
animal collagen. Gelatin used in the manufacture of capsule without interrupting the flow of carbon dioxide, introduce
shells or as a pharmaceutical aid in the manufacture of tablets through the funnel 25 g of the substance under examination
may contain suitable antimicrobial agents. and 80 ml of 2 M hydrochloric acid. Boil for 1 hour, disconnect
Description. Light amber to faintly yellow, translucent flakes, the absorption tube and stop the flow of carbon dioxide. Wash
sheets, shreds, powder or granules; odour, slight. Stable in air the contents of the absorption tube into a 250-ml conical flask,
but is subject to microbial decomposition when moist or in heat on a water-bath for 15 minutes and allow to cool. Titrate
solution. with 0.1 M sodium hydroxide using a 0.1 per cent w/v solution
542
IP 2007 HARD GELATIN CAPSULE SHELLS
of bromophenol blue in ethanol (20 per cent) as indicator, one end of which is rounded and closed and the other, open.
until the colour changes from yellow to violet-blue. Shapes other than cylindrical can also be formed as per
1 ml of 0.1 M sodium hydroxide is equivalent to 0.003203 g of requirements. The two pieces are uncoloured or coloured; if
sulphur dioxide. coloured, of identical or different colours; transparent or
opaque, partially or completely and printed or unprinted or
Microbial contamination (2.2.9). Total microbial count, not bear other surface markings. The cap overlaps the body and
more than 1000 per g; 1 g is free from Escherichia coli; 10 g is maintains a tight friction closure. The closure may be
free from Salmonellae. strengthened by suitable means.
Ash. Not more than 3.25 per cent, determined by the following The shells are of various sizes, usually designated by different
method. Weigh accurately 5.0 g, add about 2 g of liquid paraffin numbers, 5 being the smallest and 000 the biggest. Shells of
(to avoid loss due to swelling) and incinerate at a temperature sizes 0 to 4 are commonly used. Shells of special shapes,
not exceeding 500° until free from carbon. Cool and weigh. sizes, lengths and designations are also available. The shells
Loss on drying (2.4.19). Not more than 16.0 per cent, are smooth and uniform in size, shape and colour. Guidelines
determined by the following method. Weigh accurately about on dimensions in respect of different sizes of commonly used
1.0 g in a stainless steel dish weighing about 25 g and with a capsules are given in chapter 5.8.
diameter of 70 mm and a height of 15 mm, fitted with a cover.
Identification
Add 10 ml of water and allow to soak. Heat on a water-bath to
form a homogeneous solution and continue heating until most Boil one capsule shell with 20 ml of water, allow to cool and
of the water has evaporated. Dry for 2 hours at 105° and for centrifuge. To 5 ml of the supernatant liquid add 1 ml of picric
further periods of 30 minutes until two successive weighings acid solution and to another 5 ml add 1 ml of tannic acid
do not differ by more than 1 mg (Do not powder sheet gelatin solution; a precipitate is produced in each case.
while preparing for this test).
Tests
Gelatin intended for use in the preparation of pessaries and
suppositories complies with the following additional Odour. Keep 100 capsule shells in a well-closed bottle for 24
requirement. hours at a temperature between 30° and 40°; the shells do not
develop any foreign odour.
Jelly strength (2.4.18). Between 150 and 250 g. (The exact
jelly strength may be negotiated between the manufacturer NOTE - In order to ensure that the quality of the shells is not
and user on the basis of end use of the gelatin). affected by temperature and humidity, the capsule shells
should be conditioned at a temperature of 25° ± 2° and a
Storage. Store protected from moisture. relative humidity of 50 ± 5 per cent for not less than 12 hours
Labelling. The label states, where applicable, that the material before conducting the test for Average weight.
is suitable for the preparation of pessaries and suppositories Average weight. Weigh 100 capsule shells and determine the
and, if so, the jelly strength. average weight of a capsule. The average weight is within
± 10 per cent of the target weight shown in Table 1. (As sizes
0 to 4 are commonly used, detailed requirements are included
Hard Gelatin Capsule Shells for these sizes only. Requirements for other sizes may be
decided upon mutually between the manufacturer of the Hard
Hard Gelatin Capsule Shells are soluble containers for Gelatin Capsule Shells and the user).
incorporation of drugs, usually in the form of powders, pellets
Table 1 - Target weight of capsules
or granules, and are commonly intended for oral
administration. The shells are acted upon by digestive fluids Size Target weight (mg)
and the filled contents are released. They are composed of 0 96
gelatin, water and additives such as plasticizers, humectants, 1 76
surfactants, dispersing agents, flavouring agents, antimicrobial
2 63
agents, sweetening agents, opacifying agents and one or more
colouring agents permitted under the Drugs and Cosmetics 3 50
Rules, 1945. Ingredients other than colouring agents and 4 40
opacifying agents comply with the standards of this Disintegration (2.5.1). 15 minutes, using discs.
Pharmacopoeia.
Microbial contamination (2.2.9). Total microbial count, not
Description. Hard Gelatin Capsule Shells (shells or cases) more than 1000 per g. 1 g is free from Escherichia coli and
consist of two cylindrical, telescoping pieces (cap and body), Salmonellae.
543
GENTAMICIN SULPHATE IP 2007
Loss on drying (2.4.19). 12.5 to 16.0 per cent, determined on Test solution. Dissolve 0.5 g of the substance under
1.0 g by drying in an oven at 105° for 4 hours or to constant examination in 100 ml of water.
weight. Reference solution. A 0.5 per cent w/v solution of gentamicin
Storage. Store protected from moisture at a temperature not sulphate RS.
exceeding 30°.
Labelling. The label states (1) the size of the capsule shells; dry the plate in air, spray with ethanolic ninhydrin solution
(2) that only permitted colours, if any, have been used; (3) the and heat at 110° for 5 minutes. The three principal spots in the
storage conditions. chromatogram obtained with the test solution correspond to
those in the chromatogram obtained with the reference
solution.
Gentamicin Sulphate B. Dissolve 10 mg in 1 ml of water and add 5 ml of a 40 per cent
w/v solution of sulphuric acid. Heat on a water-bath for
10 minutes, cool and dilute to 25 ml with water. When examined
OH O in the range 240 nm to 330 nm (2.4.7), the resulting solution
1 R2 shows no absorption maximum.
R NHCH3
R3 CH3 C. In the test for Composition of gentamicin sulphate, the four
O principal peaks in the chromatogram obtained with the test
OH
solution correspond to the four peaks in the chromatogram
NH2 + H2SO4 D. Gives reaction A of sulphates (2.3.1).

O
O
Tests
OH
NH2
dioxide-free water (solution A) is clear (2.4.1), and not more
NH2 intensely coloured than degree 6 of the appropriate range of
reference solutions (2.4.1).
Gentamycin R1 R2 R3
C1 CH3 NHCH3 H
C2 CH3 NH2 H
C1a H NH2 H
C2a H NH2 CH3 Composition of gentamicin sulphate. Determine by liquid
Test solution. Add 5 ml of methanol and 4 ml of phthalaldehyde
Gentamicin Sulphate is a mixture of the sulphates of reagent to 10 ml of a 0.1 per cent w/v solution of the substance
antimicrobial substances produced by Micromonospora under examination in water, mix, add sufficient methanol to
purpurea. produce 25 ml, heat in a water-bath at 60° for 15 minutes and
Gentamicin Sulphate has a potency of not less than 590 µg of cool. If the solution is not used immediately, cool to 0° and
gentamicin per mg, calculated on the anhydrous basis. use within 4 hours.
Description. A white or almost white powder; hygroscopic. Reference solution. Prepare in the same manner as the test
solution but using 10 ml of a 0.1 per cent w/v solution of
Identification gentamicin sulphate RS in place of the solution of the
Tests A and B may be omitted if tests C and D are carried out.
Test C may be omitted if tests A, B and D are carried out. Chromatographic system
– a stainless steel column 10 to 12.5 cm x 4.6 to 5 mm,
A. Determine by thin-layer chromatography (2.4.17), coating packed with octadecylsilyl silica gel (5 µm),
the plate with silica gel G. – mobile phase: a solution containing 0.55 per cent w/v of
Mobile phase. The lower layer obtained by shaking together sodium heptanesulphonate monohydrate in a mixture
equal volumes of strong ammonia solution, chloroform and of 70 volumes of methanol, 25 volumes of water and
methanol and allowing to separate. 5 volumes of glacial acetic acid,
544
IP 2007 GENTAMICIN EYE DROPS

– spectrophotometer set at 330 nm, – a column 1.5 m x 4 mm, packed with porous polymer
– a 5 µl loop injector. beads (80 to 100 mesh)(such as Porapak Q),
If necessary, adjust the methanol content of the mobile phase – temperature:
so that in the chromatogram obtained with the reference column. constant at a point between 120° and 140°,
solution, the retention time of component C2 is 10 to 20 minutes inlet port and detector. 50° higher than column
and the peaks are well separated with relative retention times temperature,
of about 0.13 (reagent), 0.27 (component C1), 0.65 (component – flow rate. constant at 30 to 40 ml per minute of the carrier
C1a), 0.85 (component C2a) and 1.00 (component C2). gas.
Calculate the percentage w/w of methanol taking 0.792 g as its
Adjust the sensitivity and the volume of the reference solution
weight per ml (2.4.29) at 20°.
injected so that the height of the peak due to component C1 is
about 75 per cent of the full-scale deflection on the chart Sulphated ash (2.3.18). Not more than 1.0 per cent, determined
paper. Plot a horizontal baseline on the chromatogram from on 0.5 g.
the level portion of the curve immediately prior to the reagent Water (2.3.43). Not more than 15.0 per cent, determined on
peak. Measure the peak height above this baseline for each 0.3 g.
component. Repeat the procedure with the test solution. The
test is not valid unless the resolution between the peaks due Assay. Determine by the microbiological assay of antibiotics,
to components C2a and C2 is not less than 1.3. Method A (2.2.10), and express the result in µg of gentamicin
per mg.
From the peak heights in the chromatogram obtained with the
reference solution and the proportions of the components Gentamicin Sulphate intented for use in the manufacture of
declared for gentamicin sulphate RS, calculate the response parenteral preparations without a further appropriate
factors for components C1, C1a, C2a and C2. From these response procedure for removal of bacterial endotoxins complies with
factors and peak heights in the chromatogram obtained with the following additional requirement.
the test solution, calculate the proportions of components C1, Bacterial endotoxins (2.2.3). Not more than 1.67 Endotoxin
C1a, C2a and C2 in the substance under examination. The Units per mg of gentamicin.
proportions are within the following limits. C1, 25.0 to 50.0 per
Sterility. Complies with the test for sterility (2.2.11).
cent; C1a, 10.0 to 35.0 per cent; C2 + C2a, 25.0 to 55.0 per cent.
Storage. Store protected from moisture. If it is intended for
Sulphate. 32.0 to 35.0 per cent of SO4, calculated on the
use in the manufacture of parenteral or ophthalmic
anhydrous basis, determined by the following method.
preparations, the container should be sterile and sealed so as
Dissolve 0.25 g in 100 ml of distilled water, adjust the pH to 11
to exclude micro-organisms.
with strong ammonia solution and add 10 ml of 0.1 M barium
chloride. Titrate with 0.1 M disodium edetate using 0.5 mg of Labelling. The label states (1) the potency in terms of µg of
metalphthalein as indicator; when the colour of the solution gentamicin per mg; (2) whether or not the contents are intended
begins to change add 50 ml of ethanol (95 per cent) and for use in the manufacture of parenteral or ophthalmic
continue the titration until the violet-blue colour disappears. preparations.
Perform a blank determination and make any necessary
correction.
1 ml of 0.1 M barium chloride is equivalent to 0.009606 g of
sulphate, SO4. Gentamicin Eye Drops
Methanol. Not more than 1.0 per cent w/w, determined by gas Gentamicin Sulphate Eye Drops
chromatography (2.4.13). Gentamicin Eye Drops are a sterile solution of Gentamicin
Test solution (a). A 25 per cent w/v solution of the substance Sulphate in Purified Water.
under examination. Gentamicin Eye Drops contain not less than 90.0 per cent and
Test solution (b). A solution containing 25 per cent w/v of the not more than 120.0 per cent of the stated amount of
substance under examination and 0.25 per cent v/v of gentamicin.
1-propanol (internal standard).
Identification
Reference solution. A solution containing 0.25 per cent v/v of
the substance under examination and 0.25 per cent v/v of the A. Determine by thin-layer chromatography (2.4.6), coating
internal standard. the plate with silica gel G.
545
GENTAMICIN INJECTION IP 2007
Mobile phase. The lower layer obtained by shaking together Adjust the sensitivity and the volume of reference solution
equal volumes of strong ammonia solution, chloroform and injected so that the height of the peak due to component C1 is
methanol and allowing to separate. about 75 per cent of the full-scale deflection on the recorder.
Test solution. A volume of the eye drops containing 60 µg of Plot a horizontal baseline on the chromatogram from the level
gentamicin. portion of the curve immediately prior to the reagent peak.
Measure the peak height above this baseline for each
Reference solution. Dissolve 0.1 mg of gentamicin sulphate component. Repeat the procedure with the test solution. The
RS in a volume of water equivalent to the volume of the eye test is not valid unless the resolution between the peaks due
drops used. to components C2a and C2 is not less than 1.3.
Apply to the plate the specified volumes of each solution. From the peak heights in the chromatogram obtained with the
After development, dry the plate in air, spray with ethanolic reference solution and the proportions of the components
ninhydrin solution and heat at 110° for 5 minutes. The three declared for gentamicin sulphate RS, calculate the response
principal spots in the chromatogram obtained with the test factors for components C1, C1a, C2a and C2. From these response
solution correspond to those in the chromatogram obtained factors and peak heights in the chromatogram obtained with
with the reference solution. the test solution, calculate the proportions of components C1,
B. In the test for Composition of gentamicin sulphate, the four C1a, C2a and C2 in the eye drops. The proportion are within the
principal peaks in the chromatogram obtained with the test following limits. C1, 25.0 to 50.0 per cent; C1a, 10.0 to 35.0 per
solution correspond to the four peaks in the chromatogram cent; C2 + C2a, 25.0 to 55.0 per cent.
obtained with the reference solution. Other tests. Comply with the tests stated under Eye Drops.
Tests Assay. Determine by the microbiological assay of antibiotics
(2.2.10) on a solution prepared in the following manner.
pH (2.4.24). 6.5 to 7.5.
Dilute a volume of the eye drops containing about 15 mg of
Composition of gentamicin sulphate. Determine by liquid gentamicin to 50.0 ml with sterile phosphate buffer pH 8.0 and
chromatography (2.4.14). dilute 10.0 ml of the resulting solution to 50.0 ml with the same
Test solution. Dilute a suitable volume of the eye drops with solvent.
water to contain 0.045 per cent w/v of gentamicin. To 10 ml of Calculate the content of gentamicin in the eye drops, taking
the resulting solution add 5 ml of methanol, swirl and add 4 ml each 1000 Units found to be equivalent to 1 mg of gentamicin.
of phthalaldehyde reagent, mix, add sufficient methanol to
Labelling. The label states the quantity of active ingredient in
produce 25 ml, heat on a water-bath at 60° for 15 minutes and
terms of the equivalent amount of gentamicin.
cool.
gentamicin sulphate RS in place of the solution of the Gentamicin Injection
preparation under examination.
Gentamicin Sulphate Injection
Gentamicin Injection is a sterile solution of Gentamicin
– a stainless steel column 10 to 12.5 cm x 4.6 to 5 mm,
Sulphate in Water for Injection.
packed with octadecylsilyl silica gel (5 µm),
– mobile phase: 0.025 M sodium heptanesulphonate Gentamicin Injection contains not less than 95.0 per cent and
monohydrate in a mixture of 70 volumes of methanol, not more than 110.0 per cent of the stated amount of
25 volumes of water and 5 volumes of glacial acetic gentamicin.
acid, Description. A clear, colourless to pale-yellow solution with a
– flow rate. 1.5 ml per minute, faint odour.
– a 5 µl loop injector. Identification
If necessary, adjust the methanol content of the mobile phase
so that in the chromatogram obtained with the reference
solution the retention time of component C2 is 10 to 20 minutes
and the peaks are well separated with relative retention times Mobile phase. The lower layer obtained by shaking together
of about 0.13 (reagent), 0.27 (component C1), 0.65 (component equal volumes of strong ammonia solution, chloroform and
C1a), 0.85 (component C2a) and 1.00 (component C2). methanol and allowing to separate.
546
IP 2007 GLIBENCLAMIDE
Test solution. A volume of the injection containing 60 µg of Adjust the sensitivity and the volume of reference solution
gentamicin. injected so that the height of the peak due to component C1 is
about 75 per cent of full-scale deflection on the recorder. Plot
Reference solution. Dissolve 0.1 mg of gentamicin sulphate
a horizontal baseline on the chromatogram from the level
RS in a volume of water equivalent to the volume of the
portion of the curve immediately prior to the reagent peak.
injection used.
Measure the peak height above this baseline for each
Apply to the plate the specified volumes of each solution. component. Repeat the procedure with the test solution. The
After development, dry the plate in air, spray with ethanolic test is not valid unless the resolution factor between the peaks
ninhydrin solution and heat at 110° for 5 minutes. The three due to components C2a and C2 is not less than 1.3.
principal spots in the chromatogram obtained with the test From the peak heights in the chromatogram obtained with the
solution correspond to those in the chromatogram obtained reference solution and the proportions of the components
with the reference solution. declared for gentamicin sulphate RS, calculate the response
B. In the test for Composition of gentamicin sulphate, the four factors for components C1, C1a, C2a and C2. From these response
principal peaks in the chromatogram obtained with the test factors and peak heights in the chromatogram obtained with
solution correspond to the four peaks in the chromatogram the test solution, calculate the proportions of components C1,
obtained with the reference solution. C1a, C2a and C2 in the eye drops. The proportion are within the
following limits. C1, 25.0 to 50.0 per cent; C1a, 10.0 to 35.0 per
Tests cent; C2 + C2a, 25.0 to 55.0 per cent.
pH (2.4.24). 3.0 to 5.0. Bacterial endotoxins (2.2.3). Not more than 1.67 Endotoxin
Units per mg of gentamicin.
Composition of gentamicin sulphate. Determine by liquid
chromatography (2.4.14). Other tests. Complies with the tests stated under Parenteral
Test solution. Add 5 ml of methanol to 10 ml of a solution
prepared by diluting a suitable volume of the injection with Assay. Determine by the microbiological assay of antibiotics,
water to contain the equivalent of 0.045 per cent w/v of Method A (2.2.10), and express the result in mg of gentamicin
gentamicin, swirl and add 4 ml of phthalaldehyde reagent, per ml.
mix, add sufficient methanol to produce 25 ml, heat in a water- Calculate the content of gentamicin in the injection, taking
bath at 60° and cool. If the solution is not used immediately, each 1000 Units found to be equivalent to 1 mg of gentamicin.
cool at 0° and use within 4 hours.
Reference solution. Prepare in the same manner as the test equivalent amount of gentamicin in a suitable dose-volume.
gentamicin sulphate RS in place of the solution of the injection
under examination.
Glibenclamide
– a stainless steel column 10 to 12.5 cm x 4.6 to 5 mm, Glyburide
packed with octadecylsilane bonded to porous silica
(5 µm), O O O O
– mobile phase: 0.025 M sodium heptanesulphonate Cl S
monohydrate in a mixture of 70 volumes of methanol, NH N N
H H
25 volumes of water and 5 volumes of glacial acetic
acid, OCH3
C23H28ClN3O5S Mol. Wt. 494.0
– a 5 µl loop injector. Glibenclamide is 1-{4-[2-(5-chloro-2-
methoxybenzamido)ethyl}benzenesulphonyl}-3-
If necessary, adjust the methanol content of the mobile phase cyclohexylurea.
so that in the chromatogram obtained with the reference
solution the retention time of component C2 is 10 to 20 minutes Glibenclamide contains not less than 99.0 per cent and not
and the peaks are well separated with relative retention times more than 101.0 per cent of C23H28ClN3O5S, calculated on the
of about 0.13 (reagent), 0.27 (component C1), 0.65 (component dried basis.
C1a), 0.85 (component C2a) and 1.00 (component C2). Description. A white or almost white, crystalline powder.
547
GLIBENCLAMIDE TABLETS IP 2007
Identification Assay. Weigh accurately about 0.4 g and dissolve in 100 ml of

ethanol (95 per cent) with the aid of heat; titrate with 0.1 M
Test A may be omitted if tests B, C and D are carried out. Tests sodium hydroxide using 1 ml of dilute phenolphthalein
B, C and D may be omitted if test A is carried out. solution as indicator until a red colour is obtained.
A.Determine by infrared absorption spectrophotometry (2.4.6). 1 ml of 0.1 M sodium hydroxide is equivalent to 0.04940 g of
Compare the spectrum with that obtained with glibenclamide C23H28ClN3O5S.
RS or with the reference spectrum of glibenclamide
0.01 per cent w/v solution in 0.01 M methanolic hydrochloric
acid shows an absorption maximum at about 300 nm and a Glibenclamide Tablets
less intense maximum at about 275 nm; absorbance at about Glibenclamide Tablets contain not less than 90.0 per cent and
300 nm, about 0.63 and at about 275 nm, about 0.29. not more than 110.0 per cent of the stated amount of
C. In the test for Related substances, the principal spot in the glibenclamide, C23H28ClN3O5S.
that in the chromatogram obtained with reference solution (a). Identification
D. Dissolve 20 mg in 2 ml of sulphuric acid (96 per cent w/w); A. When examined in the range 230 nm to 360 nm (2.4.7), the
the solution is colourless and exhibits a blue fluorescence in solution obtained in the Assay shows an absorption maximum
ultraviolet light at 365 nm. Dissolve about 0.1 g of chloral at about 300 nm and a less intense maximum at about 275 nm.
hydrate in the solution; within 5 minutes the colour changes B. In the test for Related substances, the principal spot in the
to deep yellow and after about 20 minutes a brownish tinge is chromatogram obtained with the test solution corresponds to
produced. that in the chromatogram obtained with reference solution (a).
Tests Tests
Appearance of solution. A 1.0 per cent w/v solution in ethanol Related substances. Determine by thin-layer chromatography
(95 per cent), prepared with the aid of heat, is clear (2.4.1), (2.4.17), coating the plate with silica gel GF254.
volumes of cyclohexane, 5 volumes of glacial acetic acid
and 5 volumes of ethanol (95 per cent).
volumes of cyclohexane, 5 volumes of glacial acetic acid
containing 20 mg of Glibenclamide with four quantities, each
of 5 ml, of a mixture of 20 volumes of dichloromethane and 10
Test solution. Dissolve 0.2 g of the substance under volumes of acetone, evaporate the combined extracts to
examination in 10 ml of a mixture of equal volumes of dryness at a pressure of 2 kPa and at a temperature not
dichloromethane and methanol. exceeding 40° and dissolve the residue in 4 ml of a mixture of
Reference solution (a). A 2 per cent w/v solution of equal volumes of chloroform and methanol.
glibenclamide RS in the same solvent mixture. Reference solution (a). A 0.5 per cent w/v solution of
Reference solution (b). Dilute 1 volume of reference solution glibenclamide RS in the same solvent mixture.
(a) to 200 volumes with the same solvent mixture. Reference solution (b). Dilute 2 ml of reference solution (a) to
Apply to the plate 10 µl of each solution. After development, 100 ml with the same solvent mixture.
dry the plate in air and examine in ultraviolet light at 254 nm. Apply to the plate 10 µl of each solution. After development,
Any secondary spot in the chromatogram obtained with the dry the plate in air and examine in ultraviolet light at 254 nm.
test solution is not more intense than the spot in the Any secondary spot in the chromatogram obtained with the
chromatogram obtained with reference solution (b). test solution is not more intense than the spot in the
Heavy metals (2.3.13). 1.0 g complies with the limit test for chromatogram obtained with reference solution (b).
heavy metals, Method B (20 ppm). Uniformity of content. Comply with the test stated under
Sulphated ash (2.3.18). Not more than 0.2 per cent. Tablets.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Powder one tablet, warm with 10 ml of 0.1 M methanolic
on 1.0 g by drying in an oven at 105°. hydrochloric acid and centrifuge. Repeat the extraction with
548
IP 2007 GLIPIZIDE
three further quantities, each of 10 ml, of 0.1 M methanolic C. Determine by thin-layer chromatography (2.4.17), coating
hydrochloric acid. Cool the combined extracts, add sufficient the plate with silica gel GF254.
0.1 M methanolic hydrochloric acid to produce 50.0 ml and
Solvent mixture. Equal volumes of methanol and methylene
chloride.
C23H28ClN3O5S taking 63 as the specific absorbance at 300 nm. Mobile phase. A mixture of 25 volumes of anhydrous formic
acid, 25 volumes of ethyl acetate and 50 volumes of methylene
Other Tests. Comply with the tests stated under Tablets.
chloride.
examination in 100 ml of solvent mixture.
Glibenclamide and shake with 40 ml of 0.1 M methanolic
hydrochloric acid, heat gently and centrifuge. Repeat the Reference solution. A 0.10 per cent w/v solution of glipizide
extraction with three further quantities, each of 20 ml, of 0.1 M RS in solvent mixture.
methanolic hydrochloric acid. To the combined extracts add
sufficient 0.1 M methanolic hydrochloric acid to produce
dry the plate in air and examine in the ultraviolet light at 254
200.0 ml and measure the absorbance of the resulting solution
nm. The principal spot in the chromatogram obtained with the
at the maximum at about 300 nm (2.4.7), using 0.1 M methanolic
test solution corresponds to the principal spot in the
hydrochloric acid heated to the same degree as the blank.
Calculate the content of C23H28ClN3O5S taking 63 as the specific
Tests
(2.4.14).
Glipizide Test solution. Dissolve 25 mg of the substance under
O O O
glipizide RS in mobile phase.
S
O N N Reference solution (b). Dilute 1 ml of reference solution (a) to
N H H
N
H Chromatographic system
H3 C N – a stainless steel column 15 cm x 4.6 mm packed with
octadecylsilyl silica (5 µm),
C21H27N5O4S Mol. Wt. 445.5 – mobile phase: a mixture of 17 volumes of acetonitrile
Glipizide is 1-cyclohexyl-3-[[4-[2-[[(5-methylpyrazine-2- and 83 volumes of a 0.35 per cent w/v of dipotassium
yl)carbonyl]amino]ethyl]phenyl]sulphonyl]urea hydrogen phosphate, adjusted to pH 8.0 with
orthophosphoric acid,
Glipizide contains not less than 98.0 per cent and not more – flow rate. 1 ml per minute.
than 102.0 per cent of C21H27N5O4S, calculated on the dried – spectrophotometer set at 274 nm,
basis. – a 50 µl loop injector.
Description. A white or almost white, crystalline powder. Inject reference solution (a). Test is not valid unless the column
Identification
Test A may be omitted if tests B and C are carried out. Tests B Inject the test solution and reference solution (b). In the
and C may be omitted if test A is carried out. chromatogram obtained with the test solution, the area of any
A. Determine by infrared absorption spectrophotometry (2.4.6). secondary peak is not more than 0.5 times the area of the peak
Compare the spectrum with that obtained with glipizide RS. in the chromatogram obtained with reference solution (b) (0.5
B. When examined in the range 220 to 350 nm (2.4.7), a 0.002
not more than the area of the peak in the chromatogram
per cent solution in methanol, shows two maxima, at about
226 nm and 274 nm. The ratio of the absorbance at 226 nm to
that at about 274 nm, 2.0 to 2.4 . Sulphated ash (2.3.18). Not more than 0.2 per cent.
549
GLIPIZIDE TABLETS IP 2007
Loss on drying (2.4.19). Not more than 0.5 per cent, determined ethyl]benzenesulphonamide) in a mixture of equal volumes of
on 1 g by drying in an oven at 105º. dichloromethane and methanol.
Assay. Weigh accurately about 0.4 g, dissolve in 50 ml of Apply to the plate 20 µl of each solution. After development,
dimethylformamide, add 0.2 ml of quinaldine red solution. dry the plate in air and examine in the ultraviolet light at
Titrate with 0.1 M lithium methoxide until the colour changes 254 nm. Any spot corresponding to glipizide impurity A in the
from red to colourless. chromatogram obtained with test solution is not more intense
1 ml of 0.1 M lithium methoxide is equivalent to 0.04455 g of
solution (c) (0.5 per cent). Any other secondary spot is not
C21H27N5O4S.
Storage. Store protected from moisture. with reference solution (a) (0.5 per cent) and not more than
two such spots are more intense than the spot in the
Glipizide Tablets cent).
Glipizide Tablets contain not less than 90.0 per cent and not Assay. Weigh and powder 20 tablets. Weigh accurately a
more than 110.0 per cent of the stated amount of glipizide, quantity of the powder containing 15 mg of Glipizide, dissolve
C21H27N5O4S. in 30 ml of methanol with gentle heating on a water bath, cool
and add sufficient methanol to produce 50.0 ml. Filter and
Identification dilute 5.0 ml of the filtrate to 50.0 ml with methanol . Measure
the absorbance of the resulting solution at 274 nm (2.4.7).
A. Shake a quantity of the powdered tablets containing 25 mg Calculate the content of C21H27N5O4S taking 237 as the specific
of Glipizide with 10 ml of dichloromethane for 5 minutes, filter, absorbance at 274 nm.
dry the filtrate with anhydrous sodium sulphate, filter again
and evaporate the filtrate to dryness. The residue complies
2-Deoxy-D-Glucose
Compare the spectrum with that obtained with glipizide RS.
HO
final solution obtained in the assay shown are absorption O
maximum at about 226 nm and 274 nm. OH OH
Tests OH
Related substances. Determine by thin-layer chromatography C6H12O5 Mol. Wt. 164.2

(2.4.17). 2-Deoxy-D-Glucose is 2-deoxyGlucose
Mobile phase. A mixture of 20 volumes of ethyl acetate, 2-Deoxy-D-Gluocse contains not less than 97.0 per cent and
20 volumes of anhydrous formic acid and 40 volumes of not more than 103.0 per cent of C6H12O5, calculated on the
dichloromethane. dried basis.
Test solution. Extract a quantity of powdered tablet containing Description. A white to off white powder.
0.1 g of Glipizide with four 10 ml quantities of acetone,
evaporate the combined extracts to dryness under reduced Identification
pressure at a temperature not exceeding 30° and dissolve the
A. Determine by infrared absorption spectrophotmetry (2.4.6).
residue in sufficient of a mixture of equal volumes of
Compare with spectrum with that obtained with 2-deoxy-D-
dichloromethane and methanol to produce 5 ml.
glucose RS.
Reference solution (a). Dilute 1 volume of test solution to
200 volumes with a mixture of equal volumes of
Reference solution (b). Dilute 1 volume of test solution to
500 volumes with the same solvent mixture. Tests
Reference solution (c). A 0.010 per cent w/v solution of Specific optical rotation (2.4.22). + 44.0° to + 48.0°, determined
glipizide impurity A RS (4-[2-(5-methylpyrazine-2-carboxamido) in a 1.0 per cent w/v solution in water.
550
IP 2007 GLYCERIN
Test solution. Dissolve 0.5 g of the substance under Water (2.3.43). Not more than 1 per cent, determined on 0.2 g.
examination in 5 ml of dimethyl sulpoxide in a 20-ml headspace Sulphated ash (2.3.18). Not more than 0.2 per cent.
vial.
Reference stock solution. Weigh about 44.5 mg of toluene,
dilute to 50 ml with dimethyl sulphoxide. Test solution. Dissolve 0.2 g of the substance under
examination in 100.0 ml of the mobile phase.
Reference solution. Weigh about 25 mg of isopropyl alcohol
and 15 mg of methanol in to the 50 ml volumetric flask, add Reference solution. A 0.2 per cent w/v solution of 2-deoxy-D-
5 ml of reference stock solution and make up to the volume glucose RS in the mobile phase.
with dimethyl sulphoxide. Take 5 ml in a 20-ml headspace vial. Chromatographic system
Chromatographic system – a stainless steel column 25 cm × 4.6 mm, packed with
– a capillary column 30 m × 0.53 mm, packed with 6 per rigid spherical etyrene- divinyl benzenecopolymer (5 to
cent cynopropylphenyl and 94 per cent 10 µm),
dimethylpolysiloxane, – mobile phase: a mixture of 55 volumes of water and
– temperature 45 volumes of methanol,
column 42º to 200° @ 30º per minute, – flow rate. 0.5 ml per minute,
inlet port 180º and detector 240° – detector set at refractive index,
– flow rate. 3.6 ml per minute of the helium carrier gas. – a 10 µl loop injector.
Headspace conditions Inject the reference solution. The test is not valid unless the
vial pressure 10 psi, sqample oven 85°, pressurisation relative standard deviation for replicate injections is not more
time 0.2 minute. than 2.0 per cent.
Inject 1 ml of the refereence solution. The test is not valid Inject the test solution and the reference solution.
unless the resolution between the peaks due to isopropyl Calculated the content of C6H12O5.
alcohol, the peak due to methanol and toluene is not less than
2 and the tailing factor is not more than 1.5 for each component.
Inject 1 ml of the test solution and the refereence solution. In
the chromatogram obtained with the test solution, the area of
the peaks due to isopropyl alcohol, methanol and toluene is Glycerin
not more than the area of peaks in the chromatogram obtained
Glycerol
Related substances. Determine by liquid chromatography H OH
HO OH
(2.4.14).
Test solution. Dissolve 1 g of the substance under examination C3H8O3 Mol. Wt. 92.1
in 100 ml of the mobile phase. Glycerin is propane-1,2,3-triol.
Reference solution (a). A 1.0 per cent w/v solution of 2-deoxy- Glycerin contains not less than 98.0 per cent and not more
D-glucose RS in the mobile phase. than 101.0 per cent of C3H8O3, calculated on the anhydrous
Reference solution (b). Dilute 1 ml of reference solution (a) to basis.
100 ml with the mobile phase. Description. A clear, colourless or almost colourless, syrupy
The chromatographic system described under Assay. liquid; odourless; very hygroscopic.
Inject reference solution (a). The test is not valid unless the Identification
tailing factor is not more than 2.0 and the column efficiency in
not less than 2000 theoretical plates. Test A may be omitted if tests B, C and D are carried out. Tests
chromatogram obtained with the test solution, the area of any A. To 5 ml add 1 ml of water and mix carefully. The resulting
secondary peak is not more than 0.5 times the area of the peak solution complies with the following test.
in the chromatogram obtained with reference solution (b) Determine by infrared absorption spectrophotometry (2.4.6).
(0.5 per cent) and the sum of areas of all the secondary peaks Compare the spectrum with that obtained with glycerin (85
is not more than twice the area of the peak in the chromatogram per cent) RS or with the reference spectrum of glycerin (85 per
obtained with reference solution (b) (2.0 per cent). cent).
551
GLYCERIN IP 2007
B. Mix 1 ml with 0.5 ml of nitric acid and superimpose 0.5 ml of Reference solution. A solution containing 0.05 per cent w/v of
potassium dichromate solution; a blue ring develops at the the substance under examination, 0.05 per cent w/v of ethylene
interface of the two liquids. Allow to stand for 10 minutes; the glycol and 0.05 per cent w/v of diethylene glycol.
blue colour does not diffuse into the lower layer. Chromatographic system
C. Heat 1 ml with 2 g of potassium hydrogen sulphate in an – a glass column 2 m x 3 mm, packed with 10 per cent
evaporating dish. Irritant vapours are evolved which blacken diethylene glycol succinate on acid-washed and
filter paper moistened with alkaline potassium mercuri-iodide silanised, flux-calcinated siliceous earth (such as
solution. Chromosorb WHP 80-100 mesh),
– temperature:
D. Refractive index (2.4.27). 1.470 to 1.475, determined at 20°.
column.200°,
Tests inlet port and detector. 280°,
Appearance of solution. Dissolve 50 g of the substance under – flow rate. 30 ml per minute of the carrier gas.
examination in sufficient carbon dioxide-free water to produce
100 ml (solution A). Solution A is clear (2.4.1). Dilute 10 ml of Inject 3 µl or other suitable volume of the test solution. Record
solution A to 25 ml with water. The solution is colourless the chromatogram adjusting the sensitivity so that the height
(2.4.1). of the peak due to glycerin is more than 50 per cent of full-
scale deflection. Inject the same volume of the reference
Acidity or alkalinity. To 50 ml of solution A add 0.5 ml of solution and record the chromatogram. The order of elution is
phenolphthalein solution. The solution is colourless and not ethylene glycol, diethylene glycol and glycerin.
more than 0.2 ml of 0.1 M sodium hydroxide is required to
produce a pink colour. Reserve the final solution for the test The test is not valid unless in the chromatogram obtained
for Ester. with the reference solution the resolution factor between the
peaks corresponding to diethylene glycol and glycerin is not
Heavy metals (2.3.13). Dissolve 4.0 g in 2 ml of 0.1 M less than 3.0 and the area of any secondary peak in the
hydrochloric acid and sufficient water to produce 25 ml. The chromatogram obtained with the test solution is less than the
resulting solution complies with the limit test for heavy metals, area of the peak corresponding to diethylene glycol in the
Method A (5 ppm). chromatogram obtained with the reference solution.
Iron (2.3.14). 10.0 g complies with the limit test for iron Sugars. Heat 10 ml of solution A with 1 ml of 1 M sulphuric
(4 ppm). acid on a water-bath for 5 minutes. Add 3 ml of 2 M sodium
Chlorides (2.3.12). 20.0 ml of solution A complies with the limit hydroxide (carbonate-free), mix and add dropwise 1 ml of
test for chlorides (25 ppm). freshly prepared copper sulphate solution; a clear blue
Sulphates (2.3.17). 10.0 ml of solution A complies with the limit solution is produced. Continue heating on the water-bath for
test for sulphates (30 ppm). 5 minutes; the solution remains blue and no precipitate is
produced.
Aldehydes and reducing substances. To 7.5 ml of solution A
in a glass-stoppered flask add 7.5 ml of water and 1 ml of Sulphated ash (2.3.18). Not more than 0.01 per cent, determined
decolorised pararosaniline solution, close the flask and allow on 5.0 g.
to stand for 1 hour. Any colour produced is not more intense Water (2.3.43). Not more than 2.0 per cent, determined on 1.5 g.
than that obtained in a standard prepared at the same time and Assay. Weigh accurately about 0.1 g, mix thoroughly with 45
in the same manner but using 7.5 ml of formaldehyde standard ml of water, add 25.0 ml of a 2.14 per cent w/v solution of
solution (5 ppm CH2O) in place of solution A. The test is not sodium periodate and 1.0 ml of 1 M sulphuric acid. Allow the
valid unless the standard solution is pink. mixture to stand protected from light for 15 minutes. Add 5 ml
Ester. Add 0.1 M sodium hydroxide to the solution reserved of a 50 per cent w/v solution of ethylene glycol, allow to stand
in the test for Acidity or alkalinity until a total of 10.0 ml has protected from light for 20 minutes and titrate with 0.1 M
been added and boil under a reflux condenser for 5 minutes. sodium hydroxide using 0.5 ml of phenolphthalein solution
Cool, add 0.5 ml of phenolphthalein solution and titrate with as indicator. Repeat the procedure without the substance under
0.1 M hydrochloric acid. Not less than 8.0 ml of 0.1 M examination. The difference between the titrations represents
hydrochloric acid is required to decolorise the solution. the amount of sodium hydroxide required by the test
Ethylene glycol, diethylene glycol and related substances. substance.
Determine by gas chromatography (2.4.13). 1 ml of 0.1 M sodium hydroxide is equivalent to 0.00921 g of
Test solution. A 10.0 per cent w/v solution of the substance C3H8O3.
under examination. Storage. Store protected from moisture.
552
IP 2007 CONCENTRATED GLYCERYL TRINITRATE SOLUTION
Glyceryl Monostearate dichloromethane solution (solution A). Filter the combined

aqueous extracts through a filter paper moistened with water,
Monostearin wash the filter with two quantities, each of 5 ml, of water and
Glyceryl Monostearate is a mixture of monoglycerides of stearic dilute the combined filtrate and washings to 100.0 ml with
and palmitic acids, together with variable quantities of di- and water (solution B).
triglycerides. For monoglycerides — Filter solution A through a cotton
Glyceryl Monostearate contains not less than 35.0 per cent of wool plug. Wash the separating funnel and the filter with
monoglycerides, calculated as glyceryl monostearopalmitate, three quantities, each of 5 ml, of dichloromethane. Dilute the
C20H40O4, and not more than 7.0 per cent of free glycerin combined filtrate and washings to 100.0 ml with
C3H8O3, both calculated on the anhydrous basis. dichloromethane. To 25.0 ml of this solution add 25.0 ml of
periodic-acetic acid solution, shake cautiously, allow to stand
Description. A white or almost white, hard, waxy mass or at 25° to 30° for 30 minutes, add 100 ml of water and 12 ml of
unctuous powder or flakes; almost odourless. potassium iodide solution. Titrate the liberated iodine with
0.1 M sodium thiosulphate using 1 ml of starch solution as
Identification
indicator. Repeat the determination using 25 ml of
A. Heat 1 g with 2 g of potassium bisulphate in an evaporating dichloromethane instead of 25.0 ml of the solution under
dish. Irritant, lachrymatory fumes are evolved which darken examination. The difference between the titrations represents
filter paper impregnated with alkaline potassium mercuri- the amount of sodium thiosulphate required.
iodide solution. 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.00172 g
B. Heat 2.5 g with 40 ml of ethanolic potassium hydroxide of monoglycerides, calculated as glyceryl monostearo-
solution for 30 minutes on a water-bath under a reflux palmitate, C20H40O4.
condenser. Add 30 ml of water, evaporate the ethanol, acidify The quantity of 0.1 M sodium thiosulphate used in the assay
the hot mixture with 15 ml of dilute hydrochloric acid, cool is not less than 85 per cent of the quantity of sodium
and extract with 50 ml of ether. Wash the ether layer with two thiosulphate used in the blank assay.
quantities, each of 10 ml, of a 20 per cent w/v solution of
sodium chloride, dry the ether extract over anhydrous sodium For free glycerin — To 50.0 ml of solution B in a 400-ml conical
sulphate and filter. Evaporate the solvent and dry the residue flask fitted with a ground-glass stopper add 25.0 ml of periodic-
under reduced pressure. Melt the residue and fill one or two acetic acid solution, shake cautiously, allow to stand at 25°
capillary tubes (for the determination of melting range) and to 30° for 30 minutes, add 100 ml of water and 12 ml of potassium
allow to stand for 24 hours in a desiccator. Carry out the iodide solution. Titrate the liberated iodine with 0.1 M sodium
determination of melting range by Method II (2.4.21); the thiosulphate using 1 ml of starch solution as indicator. Repeat
residue melts at 54° to 64°. the determination using 50 ml of water instead of 50 ml of the
solution under examination. The difference between the
Tests titrations represents the amount of sodium thiosulphate
required.
Acid value (2.3.23). Not more than 5.0, determined on 0.5 g
dissolved in 50 ml of a mixture of equal volumes of ethanol 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.0023 g of
(95 per cent) and ether. glycerin, calculated as C3H8O3.
Saponification value (2.3.37). 155 to 170. Storage. Store protected from light.
Iodine value (2.3.28). Not more than 5.0 (iodine bromide
method).
Concentrated Glyceryl Trinitrate
g dissolved in a mixture of 10 ml of anhydrous methanol and Solution
10 ml of anhydrous chloroform. Concentrated Nitroglycerin Solution
Concentrated Glyceryl Trinitrate Solution is a solution of
dichloromethane in a glass-stoppered separating funnel. Add
propane-1,2,3-triol trinitrate in Ethanol (95 per cent).
25 ml of water and shake vigorously for 1 minute. Allow the
layers to separate (if an emulsion is formed, add a few drops of Concentrated Glyceryl Trinitrate Solution contains not less
glacial acetic acid). Repeat the extraction with three further than 9.0 per cent w/v and not more than 11.0 per cent w/v of
quantities, of 25, 20 and 20 ml, of water and reserve the C3H5N3O9.
553
GLYCERYL TRINITRATE TABLETS IP 2007
CAUTION — Undiluted glyceryl trinitrate can be exploded solution. Any secondary spot in the chromatogram obtained
by percussion or excessive heat. Proper precautions should with the test solution is not more intense than the spot in the
be exercised in handling it and only exceedingly small chromatogram obtained with the reference solution.
amounts should be isolated. Assay. Dilute 1.0 ml to 50.0 ml with a 90 per cent v/v solution of
Description. A clear, colourless to pale yellow solution. glacial acetic acid and dilute 10.0 ml of this solution to 100.0
ml with the same solvent. To 1.0 ml of the resulting solution
Identification add 2 ml of phenoldisulphonic acid solution, mix and allow to
stand for 15 minutes. Add 8 ml of water, mix well, allow to cool
Carry out the procedure described under Related substances
and add slowly, with swirling, 10 ml of strong ammonia
but using the following solutions.
solution. Cool and dilute to 20.0 ml with water. Measure the
Mobile phase. Toluene. absorbance of the resulting solution at about 405 nm (2.4.7),
Test solution. Dilute the substance under examination with using as the blank 1 ml of a 90 per cent v/v solution of glacial
acetone to contain 0.05 per cent w/v of glyceryl trinitrate. acetic acid treated in the same manner, beginning at the words
“add 2 ml of phenoldisulphonic acid solution,” Dissolve
Reference solution. Extract one powdered glyceryl trinitrate 0.1335 g of potassium nitrate previously dried at 105° in water
tablet 0.5 mg RS with 1 ml of acetone and centrifuge. to produce 50.0 ml; to 10.0 ml add sufficient glacial acetic
The principal spot in the chromatogram obtained with the test acid to produce 100.0 ml. Using 1.0 ml of this solution, repeat
solution corresponds to that in the chromatogram obtained the procedure beginning at the words “add 2 ml of
with the reference solution. phenoldisulphonic acid solution,....”.
B. To 1 ml add 200 ml of ether, evaporate 6 ml of the resulting Calculate the content of C3H5N3O9 from the values of the
solution to dryness and dissolve the residue in 0.2 ml of absorbances so obtained.
sulphuric acid containing a trace of diphenylamine; an intense 1 ml of the potassium nitrate solution is equivalent to 0.0002
blue colour is produced. g of C3H5N3O9.
Tests Storage. Store protected from light at a temperature between
8° and 15°.
Inorganic nitrates. Determine by thin-layer chromatography
Glyceryl Trinitrate Tablets
of acetone and 15 volumes of glacial acetic acid Nitroglycerin Tablets; Trinitrin Tablets
Test solution. The substance under examination. Glyceryl Trinitrate Tablets contain not less than 85.0 per cent
Reference solution. A freshly prepared 0.1 per cent w/v solution glyceryl trinitrate, C3H5N3O9.
of potassium nitrate in ethanol (90 per cent).
Apply to the plate 5 µl of each solution. After development, Identification
dry the plate in a stream of air and spray with diphenylamine A. Determine by thin-layer chromatography (2.4.17), coating
solution. Any spot corresponding to potassium nitrate in the the plate with silica gel G.
intense than the spot in the chromatogram obtained with the Mobile phase. Toluene.
reference solution. Test solution. Extract a quantity of the powdered tablets
Related substances. Determine by thin-layer chromatography containing 0.5 mg of glyceryl trinitrate with 1 ml of acetone
(2.4.17), coating the plate with silica gel G. and centrifuge.
Mobile phase. A mixture of 80 volumes of toluene and 20 Reference solution. Extract one powdered glyceryl trinitrate
volumes of ethyl acetate. tablet 0.5 mg RS with 1 ml of acetone and centrifuge.
Test solution. The substance under examination. Apply to the plate 20 µl of each solution. After development,
dry the plate in a stream of air, spray with diphenylamine
Reference solution. Dilute 1 volume of the test solution to 100
solution and irradiate for 15 minutes with ultraviolet light at
volumes with acetone.
365 nm. Examine the plate in daylight. The principal spot in the
Apply to the plate 5 µl of each solution. After development, chromatogram obtained with the test solution corresponds to
dry the plate in a stream of air and spray with diphenylamine that in the chromatogram obtained with the reference solution.
554
IP 2007 GLYCINE
B. Extract a quantity of the powdered tablets containing 3 mg resulting solution add 2 ml of phenoldisulphonic acid solution,
of glyceryl trinitrate with 5 ml of ether and filter. Evaporate the mix and allow to stand for 15 minutes. Add 8 ml of water, mix
ether and dissolve the residue in 0.2 ml of sulphuric acid well, allow to cool and add slowly, with swirling, 10 ml of
containing a trace of diphenylamine; an intense blue colour strong ammonia solution. Cool and dilute to 20.0 ml with
is produced. water. Measure the absorbance of the resulting solution at
about 405 nm (2.4.7), using as the blank 1 ml of a 90 per cent v/
Tests v solution of glacial acetic acid treated in the same manner,
beginning at the words “add 2 ml of phenoldisulphonic acid
Uniformity of content. Comply with the test stated under solution,”. Dissolve 0.1335 g of potassium nitrate previously
Tablets. dried at 105° in water to produce 50.0 ml; to 10.0 ml add
Place one tablet in a centrifuge tube containing a few glass sufficient glacial acetic acid to produce 100.0 ml. Using 1.0
beads, add 5 ml of a 90 per cent v/v solution of glacial acetic ml of this solution, repeat the procedure beginning at the
acid, shake for 1 hour and centrifuge. To 1.0 ml of the resulting words “add 2 ml of phenoldisulphonic acid solution,”.
solution add 2 ml of phenoldisulphonic acid solution, mix Calculate the content of C3H5N3O9 from the values of the
and allow to stand for 15 minutes. Add 8 ml of water, mix well, absorbances so obtained.
allow to cool and add slowly, with swirling, 10 ml of strong
ammonia solution. Cool and dilute to 20.0 ml with water. 1 ml of the potassium nitrate solution is equivalent to 0.0002 g
Measure the absorbance of the resulting solution at about of C3H5N3O9.
405 nm (2.4.7), using as the blank 1 ml of a 90 per cent v/v Storage. Store protected from light and moisture in glass
solution of glacial acetic acid treated in the same manner, containers of not more than 100 tablets, at a temperature not
beginning at the words “add 2 ml of phenoldisulphonic acid exceeding 30°. The container should be closed by means of a
solution,” Dissolve 0.1335 g of potassium nitrate previously screw cap lined with aluminium or tin foil. Cotton wool wadding
dried at 105° in water to produce 50.0 ml; to 10.0 ml add sufficient or other additional packing that absorbs glyceryl trinitrate
glacial acetic acid to produce 100.0 ml. Using 1.0 ml of this should be avoided.
solution, repeat the procedure beginning at the words “add 2
ml of phenoldisulphonic acid solution,....”. Labelling. The label states that the tablets should be allowed
to dissolve slowly in the mouth.
Calculate the content of C3H5N3O9 in the tablet from the values
of the absorbances so obtained.
1 ml of the potassium nitrate solution is equivalent to 0.0002 g Glycine
of C3H5N3O9.
Aminoacetic acid
For tablets containing 400 to 600 µg — Use 1.0 ml of a
mixture of equal volumes of the potassium nitrate solution O
and glacial acetic acid in the repeat procedure.
H2 N
For tablets containing 200 to 300 µg — Use 2.0 ml of the OH
resulting solution, prepare the blank with 2.0 ml of a 90 per C2H5NO2 Mol. Wt. 75.1
cent v/v solution of glacial acetic acid and use 2.0 ml of a
mixture of 3 volumes of glacial acetic acid and 1.0 volume of Glycine is 2-aminoethanoic acid.
the potassium nitrate solution in the repeat procedure. Glycine contains not less than 98.5 per cent and not more than
For tablets containing less than 200 µg — Use 2.0 ml of the 101.5 per cent of C2H5NO2, calculated on the dried basis.
resulting solution, measure the absorbance of 2-cm layers, Description. A white, crystalline powder; odourless.
prepare the blank with 2.0 ml of a 90 per cent v/v solution of
glacial acetic acid and use 2.0 ml of a mixture of 7 volumes of Identification
glacial acetic acid and 1.0 volume of the potassium nitrate
solution in the repeat procedure.
Compare the spectrum with that obtained with glycine RS.
Other tests. Comply with the tests stated under Tablets. The Examine the substances as discs prepared using about 1 mg
test for Disintegration does not apply. for 0.4 g of potassium bromide IR.
Assay. Weigh and powder 20 tablets. Weigh accurately a B. Dissolve 50 mg in 5 ml of water, add 1 ml of sodium
quantity of the powder containing about 1 mg of glyceryl hypochlorite solution (3 per cent Cl), boil for 2 minutes, add
trinitrate, add 5 ml of a 90 per cent v/v solution of glacial 1 ml of hydrochloric acid and boil for 4 to 5 minutes. To the
acetic acid, shake for 1 hour and centrifuge. To 1.0 ml of the resulting solution add 2 ml of hydrochloric acid and 1 ml of a
555
GLYCINE IRRIGATION SOLUTION IP 2007
2 per cent w/v solution of resorcinol, boil for 1 minute, cool, Reference solution. A 0.25 per cent w/v solution of glycine
add 10 ml of water and mix. To 5 ml of this solution add 6 ml of RS.
2 M sodium hydroxide. The resulting solution is violet with a Apply to the plate 2 µl of each solution. After development,
greenish yellow fluorescence. After a few minutes the solution dry the plate at 105° for 10 minutes, spray with ninhydrin
becomes orange and then yellow and the intense fluorescence solution and heat at 105° for 2 minutes. The principal spot in
remains. the chromatogram obtained with the test solution corresponds
Tests to that in the chromatogram obtained with the reference
solution.
dioxide-free water is clear (2.4.1), and not more intensely Tests
pH (2.4.24). 4.5 to 6.5.
Heavy metals (2.3.13). 2.0 g complies with the limit test for per ml.
Other tests. Complies with the tests stated under parenteral
Chlorides (2.3.12). 2.5 g dissolved in 20 ml of water complies Preparations (Injections).
with the limit test for chlorides (100 ppm).
Assay. Dilute an accurately measured volume of the preparation
Sulphated ash (2.3.18). Not more than 0.1 per cent. under examination containing about 0.15 g of Glycine to 25 ml
Loss on drying (2.4.19). Not more than 0.5 per cent, determined with water. Add 10 ml of formaldehyde solution, previously
on 1.0 g by drying in an oven at 105° for 2 hours. adjusted to a pH of 9.0, and 0.25 ml of a mixed indicator solution
prepared by dissolving 75 mg of phenolphthalein and 25 mg
Assay. Weigh accurately about 0.15 g and dissolve in 100 ml
of thymol blue in 100 ml of ethanol (50 per cent). Titrate with
of anhydrous glacial acetic acid. Immediately after dissolution
0.1 M sodium hydroxide until the yellow colour disappears
titrate 0.1 M perchloric acid, using 0.05 ml of crystal violet
and a faint violet colour appears.
solution as indicator. Carry out a blank titration.
C2H5NO2.
C2H5NO2.
exceeding 30°.
Glycine Irrigation Solution Labelling. The label states (1) Not for Injection; (2) that the
solution should not be used if it contains visible particles.
Glycine Irrigation Solution is a sterile solution of Glycine in
Water for Injections.
Glycine Irrigation Solution contains not less than 95.0 per
cent and not more than 105.0 per cent of the stated amount of Griseofulvin
C2H5NO2. It contains no antimicrobial agent.
H3CO O OCH3
Description. A clear, colourless solution.
Identification O
H3CO O
A.Evaporate 5 ml to dryness on a water-bath and dry at 105° CH3
for one hour. The residue complies with the following test. Cl
Determine by infrared absorption spectrophotometry (2.4.6). C17H17ClO6 Mol. Wt. 352.8
Compare the spectrum with that obtained with glycine RS. Griseofulvin is (1S,6'R)-7-chloro-2',4,6-trimethoxy-6'-
B. Determine by thin-layer chromatography (2.4.17), coating methylbenzofuran-2-spiro-1-cyclohex-2'-ene-3,4'-dione
the plate with silica gel G. produced by the growth of certain strains of Penicillium
griseofulvum or obtained by any other means.
Mobile phase. A mixture of 70 volumes of 1-propanol and 30
volumes of strong ammonia solution. Griseofulvin contains not less than 97.0 per cent and not more
Test solution. Dilute a suitable volume of the preparation under than 102.0 per cent of C17H17ClO6, calculated on the dried basis.
examination with water so that the resulting solution contains Description. A white to yellowish white powder, the particles
0.25 per cent w/v of Glycine. of which are generally upto 5 µm in maximum dimension,
556
IP 2007 GRISEOFULVIN TABLETS
although larger particles, which may occasionally exceed 30 Calculate the ratio (r) of the area of the peak due to griseofulvin
µm may be present; almost odourless. to that of the peak due to the internal standard in the
chromatogram obtained with the reference solution. The ratio
Identification of the area of any peak corresponding to dechlorogriseofulvin
A. Determine by infrared absorption spectrophotometry (2.4.6). to that of the peak due to the internal standard in the
Compare the spectrum with that obtained with griseofulvin chromatogram obtained with test solution (b) is less than 0.6r.
RS. The ratio of the area of any peak corresponding to
dehydrogriseofulvin to that of the peak due to the internal
B. Dissolve about 5 mg in 1 ml of sulphuric acid and add 5 mg
standard in the chromatogram obtained with test solution (b)
of powdered potassium dichromate; a wine-red colour is
is less than 0.15r.
produced.
C. Melting range (2.4.21). 217 ° to 224°.
Tests Matter soluble in light petroleum. Not more than 0.2 per cent,
Appearance of solution. A 7.5 per cent w/v solution in determined by the following method. Extract 1 g with 20 ml of
dimethylformamide is clear (2.4.1), and not more intensely light petroleum (40° to 60°) by boiling under a reflux
coloured than reference solution YS4 (2.4.1). condenser for 10 minutes; cool, filter, wash the filter with three
quantities, each of 15 ml, of the light petroleum, evaporate the
Acidity. Suspend 0.25 g in 20 ml of ethanol (95 per cent) and
combined filtrate and washings to dryness, dry the residue at
titrate with 0.2 M sodium hydroxide using phenolphthalein
105° for 1 hour and weigh.
solution as indicator; not more than 1.0 ml is required to change
the colour of the solution. Sulphated ash (2.3.18). Not more than 0.2 per cent.
Specific optical rotation (2.4.22). +352° to +364°, determined Loss on drying (2.4.19). Not more than 1.0 per cent, determined
at 20° in a 1.0 per cent w/v solution in dimethylformamide. on 1.0 g by drying in an oven at 105°.
Related substances. Determine by gas chromatography Assay. Weigh accurately about 80 mg and dissolve in sufficient
(2.4.13). ethanol to produce 200.0 ml. Dilute 2.0 ml to 100.0 ml with
Test solution (a). Dissolve 1.0 g of the substance under ethanol and measure the absorbance of the resulting solution
examination in 100 ml of acetone. at the maximum at about 291 nm (2.4.7). Calculate the content
of C17H17ClO6 taking 686 as the specific absorbance at 291 nm.
examination and 20 mg of 9,10-diphenylanthracene (internal
standard) in 100 ml of acetone.
Reference solution. A solution containing 0.050 per cent w/v Griseofulvin Tablets
of griseofulvin RS and 0.020 per cent w/v of the internal
Griseofulvin Tablets contain not less than 95.0 per cent and
standard in acetone.
Chromatographic system griseofulvin, C17H17ClO6.
diatomaceous support (100 to 200 mesh) coated with Identification
1 per cent w/w of cyanopropylmethyl phenyl silicone
fluid, A. Extract a quantity of the powdered tablets containing
– temperature: 0.125 g of Griseofulvin with 20 ml of chloroform, add 1 g of
column. 250°, anhydrous sodium sulphate, shake and filter. Evaporate the
inlet port and detector. 270°, filtrate to dryness and dry at a pressure not exceeding 0.7 kPa
– flow rate. 60 ml per minute of the carrier gas. for 1 hour. The residue complies with the following test.
Continue the chromatography for three times the retention Determine by infrared absorption spectrophotometry (2.4.6).
time of griseofulvin. Compare the spectrum with that obtained with griseofulvin
RS.
The chromatogram obtained with test solution (a) shows a
peak due to griseofulvin (retention time about 11 minutes) B. Shake a quantity of the powdered tablets containing 80 mg
and may show a peak due to dechlorogriseofulvin (retention of Griseofulvin with 150 ml of ethanol (95 per cent) for
time about 0.6 times that of griseofulvin) and a peak due to 20 minutes. Dilute to 200 ml with ethanol (95 per cent) and
dehydrogriseofulvin (retention time about 1.4 times that of filter. Dilute 2 ml of the filtrate to 100 ml with ethanol (95 per
griseofulvin). cent). When examined in the range 230 nm to 360 nm (2.4.7),
557
GUAIPHENESIN IP 2007
the resulting solution shows absorption maxima at about 291 Calculate the ratio (r) of the area of the peak due to griseofulvin
nm and 325 nm, and a shoulder at about 250 nm. to that of the peak due to the internal standard in the
C. Dissolve about 5 mg of the powdered tablets in 1 ml of chromatogram obtained with the reference solution. The ratio
sulphuric acid and add 5 mg of powdered potassium of the area of any peak corresponding to dechlorogriseofulvin
dichromate; a wine-red colour is produced. (retention time about 0.6 times that of griseofulvin) to that of
the peak due to the internal standard in the chromatogram
Tests obtained with test solution (b) is less than 0.6r. The ratio of
the area of any peak corresponding to dehydrogriseofulvin
Related substances. Determine by gas chromatography (retention time about 1.4 times that of griseofulvin) to the area
(2.4.13). of the peak due to the internal standard is less than 0.15r.
Test solution (a). Add 60 ml of chloroform to a quantity of the Dissolution (2.5.2).
powdered tablets containing 50 mg of Griseofulvin, heat at Apparatus. No 1
60° with shaking for 20 minutes, cool and dilute to 100 ml with
Medium. 900 ml of a 4.0 per cent w/v solution of sodium lauryl
chloroform. Centrifuge and evaporate 20 ml of the clear
sulphate
supernatant liquid to about 1 ml.
(a) but adding 1 ml of a 0.1 per cent w/v solution of 9,10-
the absorbance of the filtrate, suitably diluted with methanol
diphenylanthracene (internal standard) in chloroform before
(80 per cent), at the maximum at about 291 nm (2.4.7). Calculate
diluting to 100 ml with chloroform.
the content of C 17H 17ClO 6. taking 725 as the specific
Reference solution. Dissolve 5 mg of griseofulvin RS in absorbance at the maximum at about 291 nm.
chloroform and add 2 ml of the internal standard solution and
D. Not less than 70 per cent of the stated amount of C17H17ClO6.
sufficient chloroform to produce 200 ml. Evaporate 20 ml of
the solution to about 1 ml. Other tests. Comply with the tests stated under Tablets.
Chromatographic system Assay. Weigh and powder 20 tablets. Weigh accurately a
– a glass column 1.0 m x 4 mm, packed with acid-washed quantity of the powder containing about 35 mg of Griseofulvin,
diatomaceous support (100 to 200 mesh) coated with add 60 ml of ethyl acetate. Centrifuge and transfer two
1 per cent w/w of cyanopropylmethyl phenyl silicone quantities, each of 5 ml, of the clear supernatant liquid into
fluid, separate 100-ml volumetric flasks. Add 5 ml of 2 M methanolic
– temperature: methanesulphonic acid to the first flask, allow to stand at 20°
column. 250°, for 30 minutes and dilute to 100.0 ml with methanol (solution
inlet port and detector. 270°, A). Dilute the contents of the second flask to 100.0 ml with
– flow rate. 60 ml per minute of the carrier gas. methanol (solution B). To a third volumetric flask add 5 ml of
2 M methanolic methanesulphonic acid and dilute to 100.0
Continue the chromatography for three times the retention
ml with methanol (solution C). Measure the absorbance of
time of griseofulvin.
each solution at the maximum at about 266 nm (2.4.7). Calculate
The chromatogram obtained with test solution (a) shows a the content of C17H17ClO6 from the difference between the
peak due to griseofulvin (retention time about 11 minutes) absorbance obtained with solution A and the sum of the
and may show a peak due to dechlorogriseofulvin (retention absorbances obtained with solutions B and C and from the
time about 0.6 times that of griseofulvin) and a peak due to difference obtained by repeating the experiment using 35 mg
dehydrogriseofulvin (retention time about 1.4 times that of of griseofulvin RS in place of the powdered tablets.
griseofulvin).
Calculate the ratio (r) of the area of the peak due to griseofulvin
to that of the peak due to the internal standard in the Guaiphenesin
chromatogram obtained with the reference solution. The ratio
of the area of any peak corresponding to dechlorogriseofulvin OCH3 OH
to that of the peak due to the internal standard in the O OH
chromatogram obtained with test solution (b) is less than 0.6r.
The ratio of the area of any peak corresponding to
dehydrogriseofulvin to that of the peak due to the internal
C10H14O4 Mol. Wt. 198.2
standard in the chromatogram obtained with test solution (b)
is less than 0.15r. Guaiphenesin is (RS)-3-(2-methoxyphenoxy)propane-1,2-diol.
558
IP 2007 GUAIPHENESIN
Guaiphenesin contains not less than 98.0 per cent and not Reference solution (b). A 0.2 per cent w/v solution of
more than 101.5 per cent of C10H14O4, calculated on the dried guaiphenesin RS in dichloromethane.
basis. Apply to the plate 5 µl of each solution. After development,
Description. A white or almost white, crystalline powder; dry the plate in air and spray with a mixture of equal volumes
odourless or with a slight characteristic odour. of a 1 per cent w/v solution of potassium ferricyanide, a
20 per cent w/v solution of ferric chloride hexahydrate and
Identification ethanol (95 per cent). Any secondary spot in the
Heavy metals (2.3.13). 12 ml of a solution prepared by
Compare the spectrum with that obtained with guaiphenesin
dissolving 2.0 g in 25 ml of a mixture of 9 volumes of ethanol
RS or with the reference spectrum of guaiphenesin.
(95 per cent) and 1 volume of water complies with the limit
B. In the test for Related substances, the principal spot in the test for heavy metals, Method D (25 ppm).
Chlorides and monochlorohydrins. To 10 ml of a 2.0 per cent
that in the chromatogram obtained with reference
w/v solution, add 2 ml of 2 M sodium hydroxide, heat on a
solution (b).
water-bath for 5 minutes, cool and add 3 ml of 2 M nitric acid.
C. Melts at 79° to 83° (2.4.21). The resulting solution complies with the limit test for chlorides
(2.3.12) using 2.0 ml of chloride standard solution (25 ppm
Tests Cl)(250 ppm).
Appearance of solution. A 2.0 per cent w/v solution is clear Guaiacol. To 10 ml of a 2.0 per cent w/v solution add 0.1 ml of
(2.4.1), and colourless (2.4.1). ferric chloride test solution and allow to stand for 5 minutes.
pH (2.4.24). 5.0 to 7.0, determined in a 1.0 per cent w/v solution. The resulting solution is not more intensely coloured than a
mixture of 0.5 ml of CSS, 1.5 ml of FCS, 3.5 ml of CCS and 4.5 ml
of a solution of hydrochloric acid containing 1 per cent w/v
(2.4.17), using a plate prepared in the following manner. Mix
of HCl (2.4.1).
0.3 g of carbomer with 240 ml of water, allow to stand with
moderate stirring for 1 hour, adjust to pH 7 by the gradual Sulphated ash (2.3.18). Not more than 0.1 per cent.
addition, with stirring, of 2 M sodium hydroxide and add 30 g Loss on drying (2.4.19). Not more than 0.5 per cent, determined
of silica gel H. Spread a uniform layer of the suspension 0.75 on 1.0 g by drying in an oven at 60° over phosphorus pentoxide
mm thick, allow the coated plate to dry in air for 16 hours, heat at a pressure of 1.5 to 2.5 kPa for 3 hours.
at 105° for 1 hour, allow to cool and use immediately.
Assay. Weigh accurately about 50 mg and dissolve in 10 ml of
Mobile phase. A mixture of 80 volumes of carbon tetrachloride
water. Add 20 ml of sodium periodate solution and allow to
stand for 10 minutes. Add 25.0 ml of sodium arsenite solution
Test solution (a). Dissolve 0.2 g of the substance under and 1 ml of a 16.6 per cent w/v solution of potassium iodide,
examination in 10 ml of dichloromethane. allow to stand for 10 minutes and titrate with 0.05 M iodine
Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with using 2 ml of starch solution as indicator. Repeat the procedure
dichloromethane. without the substance under examination. The difference
between the titrations represents the amount of iodine required.
substance under examination in dichloromethane. 1 ml of 0.05 M iodine is equivalent to 0.009911 g of C10H14O4.
559
H
Haloperidol ....
Haloperidol Injection ....
Haloperidol Oral Solution ....
Haloperidol Tablets ....
Heparin Sodium ....
Heparin Injection ....
Histamine Phosphate ....
Histamine Phosphate Injection ....
Homatropine Hydrobromide ....
Homatropine Eye Drops ....
Hyaluronidase ....
Hyaluronidase Injection ....
Hydralazine Hydrochloride ....
Hydralazine Injection ....
Hydrochloric Acid ....
Dilute Hydrochloric Acid ....
Hydrochlorothiazide ....
Hydrochlorothiazide Tablets ....
Hydrocortisone ....
Hydrocortisone Acetate ....
Hydrocortisone Eye Ointment ....
Hydrocortisone Acetate Injection ....
Hydrocortisone Hemisuccinate ....
Hydrocortisone Sodium Succinate Injection ....
Hydrogen Peroxide Solution (20 Vol) ....
Hydrogen Peroxide Solution (100 Vol) ....
Hydroxocobalamin ....
Hydroxocobalamin Injection ....
Hydroxyprogesterone Hexanoate ....
Hydroxyprogesterone Injection ....
561
Hydroxypropyl Cellulose ....

Hydroxypropylmethylcellulose ....
Hyoscine Butylbromide ....
Hyoscine Butylbromide Injection ....
Hyoscine Butylbromide Tablets ....
Hyoscine Hydrobromide ....
Hyoscine Hydrobromide Injection ....
Hyoscine Hydrobromide Tablets ....
562
IP 2007 HALOPERIDOL INJECTION

Haloperidol examination in 10 ml of chloroform.
F Test solution (b). Dilute 5 ml of test solution (a) to 50 ml with
chloroform.
N Reference solution (a). A solution containing 0.005 per cent
Cl
O w/v of the substance under examination in chloroform.
OH Reference solution (b). A 0.1 per cent w/v solution of
haloperidol RS in chloroform.
C21H23ClFNO2 Mol. Wt. 375.9
Haloperidol is 4-[4-(4-chlorophenyl)-4-hydroxypiperidino]- dry the plate in air and spray with dilute potassium
4′-fluorobutyrophenone. iodobismuthate solution. Any secondary spot in the
Haloperidol contains not less than 99.0 per cent and not more chromatogram obtained with test solution (a) is not more
than 101.0 per cent of C21H23ClFNO2, calculated on the dried intense than the spot in the chromatogram obtained with
basis. reference solution (a). The test is not valid unless the
chromatogram obtained with reference solution (a) shows a
Description. A white to faintly yellowish, amorphous or distinct and clearly visible spot.
microcrystalline powder; odourless.
Identification Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Test A may be omitted if tests B, C and D are carried out. Tests on 1.0 g by drying in an oven at 60° at a pressure not exceeding
B, C and D may be omitted if test A is carried out. 0.1 kPa for 3 hours.
A. Determine by infrared absorption spectrophotometry (2.4.6). Assay. Weigh accurately about 0.2 g and dissolve in 25 ml of
Compare the spectrum with that obtained with haloperidol anhydrous glacial acetic acid. Titrate with 0.05 M perchloric
RS or with the reference spectrum of haloperidol. acid, using 0.2 ml of 1-naphtholbenzein solution as indicator
and titrating until the colour changes from orange-yellow to
B. When examined in the range 230 nm to 360 nm (2.4.7), a yellowish green. Carry out a blank titration.
0.0015 per cent w/v solution in a mixture of 90 volumes of
methanol and 10 volumes of 0.1 M hydrochloric acid shows 1 ml of 0.05 M perchloric acid is equivalent to 0.01879 g
an absorption maximum at about 245 nm; absorbance at about C21H23ClFNO2.
245 nm, about 0.49 to 0.53. Storage. Store protected from light.
that in the chromatogram obtained with reference solution (b). Haloperidol Injection
D. Determine by the oxygen-flask method (2.3.34), using 20 Haloperidol Injection is a sterile solution of Haloperidol in
mg of the substance under examination and 5 ml of 1.25 M Lactic Acid diluted with Water for Injections.
sodium hydroxide as the absorbing liquid. When the process
is complete, dilute to 10 ml with water; the resulting solution Haloperidol Injection contains not less than 90.0 per cent and
complies with the following tests. not more than 110.0 per cent of the stated amount of
haloperidol, C21H23ClFNO2.
(a) Add 0.1 ml to a mixture of 0.1 ml of a freshly prepared
alizarin red S solution and 0.1 ml of zirconyl nitrate solution; Identification
the red colour becomes clear yellow.
A. To a volume of the injection containing 20 mg of Haloperidol
(b) Acidify 5 ml with 0.5 M sulphuric acid; the solution gives add 5 ml of water and 1 ml of 1 M sodium hydroxide and
the reactions of chlorides (2.3.1). extract with 10 ml of chloroform. Filter the chloroform extract
through absorbent cotton, evaporate the filtrate to dryness
Tests
and dry the residue at 60° at a pressure not exceeding 0.7 kPa.
Related substances. Determine by thin-layer chromatography The residue complies with the following test.
(2.4.17), coating the plate with silica gel G. Determine by infrared absorption spectrophotometry (2.4.6).
Mobile phase. A mixture of 80 volumes of chloroform, 10 Compare the spectrum with that obtained with haloperidol
volumes of glacial acetic acid and 10 volumes of methanol. RS or with the reference spectrum of haloperidol.
563
HALOPERIDOL ORAL SOLUTION IP 2007
B. When examined in the range 230 nm to 360 nm (2.4.7), the Description. A clear, colourless solution.
maximum only at about 245 nm. Identification
A. To a volume of the oral solution containing 20 mg of
Tests Haloperidol, add 1 ml of 1 M sodium hydroxide, extract with
pH (2.4.24). 2.8 to 3.6. 10 ml of chloroform, filter and evaporate the filtrate to dryness.
The residue complies with the following test.
(2.4.17), coating the plate with silica gel G. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with haloperidol
Mobile phase. A mixture of 80 volumes of chloroform, 10 RS treated in the same manner or with the reference spectrum
volumes of glacial acetic acid and 10 volumes of methanol. of haloperidol.
Test solution. The injection under examination. B. When examined in the range 230 nm to 360 nm (2.4.7), the
Reference solution (a). Dilute 1 volume of the injection to 100 final solution obtained in the Assay shows an absorption
volumes with methanol. maximum only at about 245 nm.
Reference solution (b). Dilute 1 volume of the injection to 200 Tests
volumes with methanol.
pH (2.4.24). 3.5 to 4.5.
Apply to the plate a volume of the injection containing 0.1 mg
of Haloperidol and the same volume of the reference solutions.
After development, dry the plate in air and spray with dilute
potassium iodobismuthate solution. Any secondary spot in Mobile phase. A mixture of 92 volumes of dichloromethane, 8
the chromatogram obtained with the test solution is not more volumes of methanol and 1 volume of strong ammonia
intense than the spot in the chromatogram obtained with solution.
reference solution (a) and not more than one such spot is Test solution. Dilute the oral solution if necessary with
more intense than the spot in the chromatogram obtained methanol to contain 0.1 per cent w/v of Haloperidol.
with reference solution (b).
Other tests. Complies with the tests stated under Parenteral 100 volumes with methanol.
Reference solution (b). Dilute 1 volume of the test solution to
Assay. To an accurately measured volume of the injection 200 volumes with methanol.
containing about 10 mg of Haloperidol add 8 ml of water and Apply to the plate 50 µl of each solution. After development,
10 ml of 1 M hydrochloric acid. Extract with successive dry the plate in air and spray with dilute potassium
quantities of 25, 25, 10 and 10 ml of ether. Wash the combined iodobismuthate solution. Any secondary spot in the
ether extracts with 10 ml of water, combine the aqueous layers chromatogram obtained with the test solution is not more
and remove the ether using a rotary evaporator. Add sufficient intense than the spot in the chromatogram obtained with
water to produce 100.0 ml and dilute 10.0 ml to 100.0 ml with reference solution (a) and not more than one such spot is
methanol. Measure the absorbance of the resulting solution more intense than the spot in the chromatogram obtained
at the maximum at about 245 nm (2.4.7). Calculate the content with reference solution (b).
of C21H23ClFNO2 taking 346 as the specific absorbance at 245
nm. Other tests. Complies with the tests stated under Oral Liquids.
Storage. Store protected from light. Assay. To an accurately measured volume of the oral solution
containing about 10 mg of Haloperidol add 8 ml of water and
10 ml of 1 M hydrochloric acid. Extract with successive
quantities of 25, 25, 10 and 10 ml of ether. Wash the combined
Haloperidol Oral Solution ether extracts with 10 ml of water, combine the aqueous layers
Haloperidol Oral Drops; Haloperidol Solution and remove the ether using a rotary evaporator. Add sufficient
water to produce 100.0 ml and dilute 10.0 ml to 100.0 ml with
Haloperidol Oral Solution is a solution of Haloperidol in Purified methanol. Measure the absorbance of the resulting solution at
Water prepared with the aid of Lactic Acid. the maximum at about 245 nm (2.4.7). Calculate the content of
Haloperidol Oral Solution contains not less than 95.0 per cent C21H23ClFNO2 taking 346 as the specific absorbance at 245 nm.
and not more than 105.0 per cent of the stated amount of Storage. Store protected from light at a temperature between
haloperidol, C21H23ClFNO2. 15° and 25°.
564
IP 2007 HEPARIN SODIUM
Haloperidol Tablets the supernatant liquid after diluting suitably with the mobile
phase if necessary.
Haloperidol Tablets contain not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of Reference solution. A solution containing 0.015 per cent w/v
haloperidol, C21H23ClFNO2. of haloperidol RS in the mobile phase.
Identification – a stainless steel column 15 cm x 5 mm, packed with
A. To a quantity of the powdered tablets containing 10 mg of
– mobile phase: a mixture of 55 volumes of a 1 per cent w/
Haloperidol add 5 ml of water and 1 ml of 1 M sodium hydroxide
v solution of ammonium acetate and 45 volumes of
and extract with 10 ml of chloroform. Filter the chloroform
acetonitrile,
extract through absorbent cotton, evaporate the filtrate to
dryness and dry the residue at 60° at a pressure not exceeding
0.7 kPa. The residue complies with the following test.
Compare the spectrum with that obtained with haloperidol Calculate the content of C21H23ClFNO2 in the tablet.
RS or with the reference spectrum of haloperidol. Other tests. Comply with the tests stated under Tablets.
B. In the test for Related substances, the principal spot in the Assay. For tablets containing more than 2 mg — Weigh and
chromatogram obtained with test solution (b) corresponds to powder 20 tablets. On the powder determine by liquid
that in the chromatogram obtained with reference solution (b). chromatography (2.4.14).
Tests Test solution. Weigh accurately a quantity of the powder

containing about 20 mg of Haloperidol, shake with 60 ml of the
Related substances. Determine by thin-layer chromatography mobile phase, place in an ultrasonic bath for 2 minutes, add
(2.4.17), coating the plate with silica gel G. sufficient quantity of the mobile phase to produce 100.0 ml.
Mobile phase. A mixture of 80 volumes of chloroform, 10 Centrifuge and use the supernatant liquid.
volumes of glacial acetic acid and 10 volumes of methanol. Reference solution. A solution containing 0.02 per cent w/v of
Test solution (a). Shake a quantity of the powdered tablets haloperidol RS in the mobile phase.
containing 10 mg of Haloperidol with 10 ml of chloroform, Chromatographic system
filter, evaporate the filtrate to dryness and dissolve the residue – a stainless steel column 15 cm x 5 mm, packed with
in 1 ml of chloroform. octadecylsilyl silica gel (5 µm),
Test solution (b). Dilute 1 volume of test solution (a) to 10 – mobile phase: a mixture of 55 volumes of a 1 per cent w/
volumes with chloroform. v solution of ammonium acetate and 45 volumes of
acetonitrile,
Reference solution (a). Dilute 1 volume of test solution (a) to – flow rate. 2 ml per minute,
200 volumes with chloroform. – spectrophotometer set at 247 nm,
Reference solution (b). A 0.1 per cent w/v solution of – a 20 µl loop injector.
haloperidol RS in chloroform. Calculate the content of C21H23ClFNO2 in the tablets.
Apply to the plate 10 µl of each solution. After development, For tablets containing 2 mg or less — Use the average of the
dry the plate in air and spray with dilute potassium 10 individual results obtained in the test for Uniformity of
iodobismuthate solution. Any secondary spot in the content.
intense than the spot in the chromatogram obtained with Storage. Store protected from light.
chromatogram obtained with reference solution (a) shows a
distinct and clearly visible spot.
Heparin Sodium
Tablets. Heparin Sodium is a preparation containing the sodium salt of
a complex organic acid present in mammalian tissues, and
Determine by liquid chromatography (2.4.14). having the characteristic property of delaying the clotting of
Test solution. Place one tablet in 10 ml of the mobile phase, shed blood. It may be obtained from the lungs or intestinal
shake in an ultrasonic bath for 2 minutes, centrifuge and use mucosa of oxen, pigs or sheep. It is prepared in conditions
565
HEPARIN SODIUM IP 2007
designed to minimise or eliminate microbial contamination and Unit. The specific activity contained in 7.7 µg of the Standard
substances lowering blood pressure. Preparation and is the same as the International Unit; 1 mg
Heparin Sodium intended for use in the manufacture of contains 130 Units.
parenteral preparations contains not less than 150 Units per Special Reagents
mg and Heparin Sodium not intended for use in the
Prepared Plasma. Collect blood from sheep or goats or human
manufacture of parenteral preparations contains not less than
volunteers directly into a vessel containing 8 per cent w/v
120 Units per mg, both calculated on the dried basis.
solution of sodium citrate in the proportion of 1 volume to
Description. A white or greyish-white powder; odourless; each 19 volumes of blood to be collected. Mix immediately by
moderately hygroscopic. gentle agitation and inversion of the vessel. Immediately
centrifuge and pool the separated plasma. To 1 ml of the pooled
Identification plasma in a clean test-tube add 0.2 ml of a 1 per cent w/v
A. It delays the clotting of freshly shed blood. solution of calcium chloride and mix. The plasma is suitable if
a solid clot forms within 5 minutes.
B. To 0.1 g in a test-tube add 0.2 g of sodium and heat
cautiously until the reaction with sodium is complete. Heat to Solution of standard preparation. Determine by preliminary
bright red heat and carefully plunge the tube and the contents trial, if necessary, approximately the minimum quantity of the
into 5 ml of water. Filter, boil the filtrate for a few minutes with Standard Preparation of heparin sodium which, when added
20 mg of ferrous sulphate. Cool, acidify with hydrochloric in 0.8 ml of saline solution, maintains fluidity in 1 ml of
acid and add 0.05 ml of ferric chloride test solution; a blue prepared plasma for 1 hour after the addition of 0.2 ml of a
colour is produced (distinction from dextran sulphate). 1 per cent w/v solution of calcium chloride. On the day of the
assay prepare a solution of the Standard Preparation such
C. The residue obtained in the test for Sulphated ash gives
that it contains in each 0.8 ml of saline solution the above-
reaction A of sodium salts (2.3.1).
determined quantity of the Standard Preparation.
Tests Test solution. Weigh accurately about 25 mg of the preparation
under examination and dissolve in sufficient saline solution
Appearance of solution. A solution containing 5000 Units per
to give a concentration of 1 mg per ml and dilute to a
ml is clear (2.4.1), and not more intensely coloured than degree
concentration estimated to correspond to that of the solution
5 of the appropriate range of reference solutions (2.4.1).
of the Standard Preparation.
Method
heavy metals, Method B (40 ppm). To very clean test-tubes (150 mm x 16 mm) add graded amounts
of the solution of standard preparation, selecting the amounts
Protein and nucleotidic impurities. Absorbance of a 0.4 per so that the largest dose does not exceed 0.8 ml and so that
cent w/v solution at about 260 nm and about 280 nm, not they correspond roughly to a geometric series in which each
greater than 0.2 and 0.15 respectively (2.4.7). step is approximately 5 per cent greater than the next lower. To
Sulphated ash (2.3.18). 30 to 43 per cent, determined on 0.2 g. each tube add sufficient saline solution to make the total
Loss on drying (2.4.19). Not more than 8.0 per cent, determined volume 0.8 ml. Add 1.0 ml of prepared plasma to each tube.
on 1.0 g by drying in an oven over phosphorus pentoxide at Then add 0.2 ml of a 1 per cent w/v solution of calcium chloride,
60° at a pressure not exceeding 0.7 kPa for 3 hours. note the time, immediately stopper each tube with a suitable
stopper and mix the contents by inverting three times in such
Assay. Determine the potency of heparin sodium by comparing a way that the entire inner surface of the tube is wet.
the concentration necessary to prevent the clotting of sheep
or goat or human plasma with the concentration of the In the same manner set up a series using the test solution,
Standard Preparation of heparin sodium necessary to give the completing the entire process of preparing and mixing the
same effect under the conditions of the following method of tubes of both the solution of standard preparation and the
assay. test solution within 20 minutes after the addition of the prepared
plasma. Exactly one hour after the addition of the calcium
Standard Preparation and Unit chloride solution, determine the extent of clotting in each tube,
Standard Preparation. The freeze-dried sodium salt of the recognising three grades between zero and full clotting.
purified active principle from bovine intestinal mucous The dilution of the test solution, which contains heparin
membranes or any other suitable preparation, the potency of sodium in the same concentration as the dilution of the solution
which has been determined in relation to the International of standard preparation, is that contained in the series of
Standard. dilutions, which show the same degree of clotting as the series
566
IP 2007 HEPARIN INJECTION
of dilutions of the solution of standard preparation. If the Tests

degree of clotting observed in the series of dilutions of the
solution of standard preparation lies between that observed pH (2.4.24). 5.5 to 8.0.
in two of the series of dilutions of the sample being examined, Bacterial endotoxins (2.2.3). Not more than 0.03 Endotoxin
the potency of the latter is estimated. If there is no such Unit per Unit of heparin.
correspondence between the degrees of clotting produced
by the solution of standard preparation and any of the
dilutions of the sample being examined, new dilutions of the
latter are prepared and assay is repeated. Assay. Determine the potency of heparin sodium by comparing
the concentration necessary to prevent the clotting of sheep
Carry out no fewer than three independent assays. Calculate
or goat or human plasma with the concentration of the
the estimated potency of the preparation being examined by
Standard Preparation of heparin sodium necessary to give the
combining the results of these assays by standard statistical
same effect.
methods.
Limits of error (P = 0.99). 90 and 110 per cent, with three Standard Preparation and Unit
determinations; 92 and 108 per cent, with four determinations. Standard Preparation. The freeze-dried sodium salt of the
Heparin Sodium intended for use in the manufacture of purified active principle from bovine intestinal mucous
parenteral preparations without a further appropriate membranes or any other suitable preparation, the potency of
procedure for the removal of bacterial endotoxins complies which has been determined in relation to the International
with the following additional requirement. Standard.
Bacterial endotoxins (2.2.3). Not more than 0.03 Endotoxin Unit. The specific activity contained in 7.7 µg of the Standard
Unit per Unit of heparin. Preparation and is the same as the International Unit; 1 mg
contains 130 Units.
Heparin Sodium intended for use in the manufacture of
parenteral preparations without a further appropriate Special Reagents
Prepared Plasma. Collect blood from sheep or goats or human
volunteers directly into a vessel containing 8 per cent w/v
Sterility. Complies with the test for sterility (2.2.11). solution of sodium citrate in the proportion of 1 volume to
Storage. Store protected from moisture in tightly-closed each 19 volumes of blood to be collected. Mix immediately by
containers, sealed so as to exclude micro-organisms. gentle agitation and inversion of the vessel. Immediately
centrifuge and pool the separated plasma. To 1 ml of the pooled
Labelling. The label states (1) the number of Units in the
plasma in a clean test-tube add 0.2 ml of a 1 per cent w/v
container; (2) the number of Units per mg; (3) the name and
solution of calcium chloride and mix. The plasma is suitable if
quantity of any added substance; (4) where applicable, that it
a solid clot forms within 5 minutes.
is sterile; (5) the source of the material (lung or mucosal).
Solution of standard preparation. Determine by preliminary
trial, if necessary, approximately the minimum quantity of the
Heparin Injection Standard Preparation of heparin sodium which, when added
in 0.8 ml of saline solution, maintains fluidity in 1 ml of
Heparin Sodium Injection prepared plasma for 1 hour after the addition of 0.2 ml of a
Heparin Injection is a sterile solution of Heparin Sodium in 1 per cent w/v solution of calcium chloride. On the day of the
Water for Injections. The pH of the solution may be adjusted assay prepare a solution of the Standard Preparation such
by the addition of a suitable alkali. that it contains in each 0.8 ml of saline solution the above-
determined quantity of the Standard Preparation.
Heparin Injection contains not less than 90.0 per cent and not
more than 110.0 per cent of the stated potency in terms of Test solution. Dilute the injection to a concentration estimated
Units per ml. to correspond to that of the solution of the Standard
Preparation.
Description. A clear, colourless or straw-coloured solution,
free from turbidity and matter which deposits on standing Method
Identification To very clean test-tubes (150 mm x 16 mm) add graded amounts
of the solution of standard preparation, selecting the amounts
A. It delays the clotting of freshly shed blood. so that the largest dose does not exceed 0.8 ml and so that
B. Gives reaction A of sodium salts (2.3.1). they correspond roughly to a geometric series in which each
567
HISTAMINE PHOSPHATE IP 2007
step is approximately 5 per cent greater than the next lower. To Histamine Phosphate is 2-(1H-imidazol-4-yl)ethylamine
each tube add sufficient saline solution to make the total diphosphate monohydrate.
volume 0.8 ml. Add 1.0 ml of prepared plasma to each tube. Histamine Phosphate contains not less than 98.0 per cent and
Then add 0.2 ml of a 1 per cent w/v solution of calcium chloride, not more than 101.0 per cent of C5H9N3,2H3PO4, calculated on
note the time, immediately stopper each tube with a suitable the anhydrous basis.
stopper and mix the contents by inverting three times in such
a way that the entire inner surface of the tube is wet. Description. Colourless, long prismatic crystals; odourless.
In the same manner set up a series using the test solution, Identification
completing the entire process of preparing and mixing the
tubes of both the solution of standard preparation and the Test A may be omitted if tests B, C and D are carried out. Tests
test solution within 20 minutes after the addition of the prepared B and C may be omitted if tests A and D are carried out.
plasma. Exactly one hour after the addition of the calcium A. Determine by infrared absorption spectrophotometry (2.4.6).
chloride solution, determine the extent of clotting in each tube, Compare the spectrum with that obtained with histamine
recognising three grades between zero and full clotting. phosphate RS.
The dilution of the test solution, which contains heparin B. In the test for Histidine, the principal spot in the
sodium in the same concentration as the dilution of the solution chromatogram obtained with test solution (b) corresponds to
of standard preparation, is that contained in the series of that in the chromatogram obtained with reference solution (a).
dilutions, which show the same degree of clotting as the series
of dilutions of the solution of standard preparation. If the C. Dissolve 0.1 g in 7 ml of water and add 3 ml of sodium
degree of clotting observed in the series of dilutions of the hydroxide solution. Dissolve 50 mg of sulphanilic acid in
solution of standard preparation lies between that observed 10 ml of water containing 0.1 ml of hydrochloric acid and 0.1
in two of the series of dilutions of the sample being examined, ml of a 10 per cent w/v solution of sodium nitrite. On mixing
the potency of the latter is estimated. If there is no such the two solutions a deep red colour is produced.
correspondence between the degrees of clotting produced D. Gives reaction A of phosphates (2.3.1).
by the solution of standard preparation and any of the
dilutions of the sample being examined, new dilutions of the Tests
latter are prepared and assay is repeated. Appearance of solution. Solution A is clear (2.4.1), and not
Carry out no fewer than three independent assays. Calculate more intensely coloured than reference solution BYS7 (2.4.1).
the estimated potency of the preparation being examined by pH (2.4.24). 3.7 to 3.9, determined in a 5.0 per cent w/v solution
combining the results of these assays by standard statistical in carbon dioxide-free water prepared from distilled water
methods. Express the result in number of Units per ml. (solution A).
Limits of error (P = 0.99). 90 and 110 per cent, with three Histidine. Determine by thin-layer chromatography (2.4.17),
determinations; 92 and 108 per cent, with four determinations. coating the plate with silica gel G.
Storage. Store at a temperature not exceeding 30°. Mobile phase. A mixture of 75 volumes of acetonitrile, 20
Labelling. The label states (1) the strength in terms of the volumes of water and 5 volumes of strong ammonia solution.
number of Units in a suitable dose-volume except that for Test solution (a). Dissolve 0.5 g of the substance under
multiple dose containers the strength is stated as the number examination in 10 ml of water.
of Units per ml; (2) the source of the material (lung or mucosal);
(3) when no antimicrobial preservative is present that the Test solution (b). Dilute 5 ml of test solution to 25 ml with
injection contains no antimicrobial preservative; (4) that any water.
portion of the contents not used at once should be discarded. Reference solution (a). A 1.0 per cent w/v solution of
histamine phosphate RS.
Histamine Phosphate w/v of DL-histidine monohydrochloride.
Histamine Acid Phosphate Reference solution (c). A mixture of equal volumes of test
solution (a) and reference solution (b).
N Apply to the plate 1 µl of each solution. After development,
HN NH2 , 2H3PO4 , H2O dry the plate in a current of air and repeat the development in
the same direction. Dry the plate in a current of air, spray with
C5H9N3,2H3PO4,H2O Mol. Wt. 325.2 ninhydrin solution and heat at 110° for 10 minutes. Any spot
568
IP 2007 HOMATROPINE HYDROBROMIDE
corresponding to histidine monohydrochloride in the tared 25 ml centrifuge tube containing a thin glass rod slightly
chromatogram obtained with test solution (a) is not more curved at the end, add 0.5 ml of nitranilic acid solution with
intense than the spot in the chromatogram obtained with continuous stirring and allow to stand for 15 minutes. Add 10
reference solution (b). The test is not valid unless the ml of ethanol (95 per cent), mix and keep at 0° for 3 hours.
chromatogram obtained with reference solution (c) shows two Centrifuge for 1 minute, dislodge any particles at the surface
clearly separated spots. and again centrifuge for 1 minute. Decant the supernatant
Sulphates (2.3.17). 3 ml of solution A diluted to 15 ml with liquid and stir the precipitate with 5 ml of ice-cold ethanol (95
distilled water complies with the limit test for sulphates (0.1 per cent). Centrifuge for 2 minutes, decant and repeat the
per cent). washing with two further quantities, each of 5 ml, of ice-cold
ethanol (95 per cent) and finally with 5 ml of ether. Smear the
Water (2.3.43). 5.0 to 6.2 per cent, determined on 0.3 g. residue over the inside of the tube by means of the glass rod
Assay. Weigh accurately about 0.14 g, dissolve in 5 ml of and dry to constant weight at 130°.
anhydrous formic acid and add 20 ml of anhydrous glacial 1 g of the residue is equivalent to 0.9529 g of
acetic acid. Titrate with 0.1 M perchloric acid, determining C5H9N3,2H3PO4,H2O.
titration. Storage. Store protected from light.

C5H9N3,2H3PO4.
Homatropine Hydrobromide
CH3
N
Histamine Phosphate Injection
Histamine Acid Phosphate Injection
H
Histamine Phosphate Injection is a sterile solution of Histamine OH
, HBr
Phosphate in Water for Injections. O
Histamine Phosphate Injection contains not less than 90.0 per O
histamine phosphate, C5H9N3,2H3PO4,H2O. C16H21NO3,HBr Mol. Wt. 356.3
Identification Homatropine Hydrobromide is (1R,3r,5S)-3-(RS)-
mandeloyloxytropane hydrobromide.
A. Evaporate a volume of the injection containing about 2 mg
of Histamine Phosphate on a water bath to dryness, dissolve Homatropine Hydrobromide contains not less than 99.0 per
the residue in 0.5 ml of water, and add 0.5 ml of sodium cent and not more than 101.0 per cent of C16H21NO3,HBr,
hydroxide. Add 2 drops of a 10 per cent w/v solution of sodium calculated on the dried basis.
nitrite and 1 ml of a solution prepared by mixing 50 mg of Description. Colourless crystals or a white, crystalline powder;
sulphanilic acid with 10 ml of water containing 2 drops of odourless.
hydrochloric acid; an orange-red colour is produced.
B. To 1 ml of the injection containing not less than 1 mg of Identification
Histamine Phosphate (concentrate a larger volume by Test A may be omitted if tests B, C and D are carried out. Tests
evaporation, if necessary), add ammonium molybdate solution B and C may be omitted if tests A and D are carried out.
dropwise; a yellow precipitate, which is soluble in ammonia,
is formed.
Compare the spectrum with that obtained with homatropine
Tests hydrobromide RS or with the reference spectrum of
homatropine hydrobromide.
pH (2.4.24). 3.0 to 6.0.
B. Dissolve 50 mg in 1 ml of water and add 2 ml of 2 M acetic
Other tests. Complies with the tests stated under Parenteral acid. Heat, add 4 ml of picric acid solution and allow to cool,
Preparations (Injections). shaking occasionally. The crystals, after washing with two
Assay. Measure accurately a volume of the injection quantities, each of 3 ml, of iced water and drying at 105° melt
containing about 10 mg of Histamine Phosphate, transfer to a at 182° to 186° (2.4.21).
569
HOMATROPINE EYE DROPS IP 2007
C. Dissolve about 10 mg in 1 ml of water, add a slight excess of Homatropine Eye Drops contain not less than 90.0 per cent
10 M ammonia and shake with 5 ml of chloroform. Evaporate and not more than 110.0 per cent of the stated amount of
the chloroform layer to dryness on a water-bath and add homatropine hydrobromide, C16H21NO3,HBr.
1.5 ml of a 2 per cent w/v solution of mercuric chloride in
ethanol (60 per cent); a yellow colour develops which becomes Identification
red on warming.
A. To a volume containing 60 mg of Homatropine
D. Gives reaction A of bromides (2.3.1). Hydrobromide add 3 ml of dilute ammonia solution, extract
with 15 ml of chloroform, dry the chloroform extract over
Tests anhydrous sodium sulphate, filter and evaporate the filtrate
Appearance of solution. A 5.0 per cent w/v solution in carbon to dryness. The residue complies with the following test.
dioxide-free water is clear (2.4.1), and not more intensely Determine by infrared absorption spectrophotometry (2.4.6).
coloured than reference solution BS8 (2.4.1). Compare the spectrum with that obtained with homatropine
pH (2.4.24). 5.0 to 6.5, determined in a 5.0 per cent w/v solution. hydrobromide RS or with the reference spectrum of
homatropine hydrobromide.
(2.4.17), coating the plate with silica gel G. B. To the residue obtained in test A, add 1.5 ml of a 2 per cent
w/v solution of mercuric chloride in ethanol (60 per cent); a
Mobile phase. A mixture of 134 volumes of ethyl acetate, 33 yellow colour is produced which becomes red on gentle
volumes of anhydrous formic acid and 33 volumes of water. warming (distinction from most other alkaloids except atropine
Test solution. Dissolve 0.4 g of the substance under and hyoscyamine).
Tests
volumes with methanol (90 per cent). Related substances. Determine by thin-layer chromatography
Apply to the plate 5 µl of each solution. After development, (2.4.17), coating the plate with silica gel G.
dry the plate at 105° until the odour of the solvent is no longer Mobile phase. A mixture of 134 volumes of ethyl acetate, 33
detectable, allow to cool and spray with dilute potassium volumes of anhydrous formic acid and 33 volumes of water.
iodobismuthate solution until spots appear. Any secondary
Test solution. Use the eye drops, diluted if necessary with
spot in the chromatogram obtained with the test solution is
water to contain 1 per cent w/v of Homatropine Hydrobromide.
not more intense than the spot in the chromatogram obtained
with the reference solution. Reference solution. Dilute 1 volume of the test solution to 200
volumes with water.
on 0.5 g. Apply to the plate 40 µl of each solution. After development,
dry the plate at 105° until the odour of the solvent is no longer
detectable, allow to cool and spray with dilute potassium
iodobismuthate solution until spots appear. Any secondary
Assay. Weigh accurately about 0.3 g, dissolve in 20 ml of spot in the chromatogram obtained with the test solution is
anhydrous glacial acetic acid and add 7 ml of mercuric acetate not more intense than the spot in the chromatogram obtained
solution. Titrate with 0.1 M perchloric acid, determining the with the reference solution. After development, dry at 105°
end-point potentiometrically (2.4.25). Carry out a blank titration. until the odour of solvent is no longer detectable, allow to
1 ml of 0.1 M perchloric acid is equivalent to 0.03563 g of cool and spray with dilute potassium iodobismuthate solution
C16H21NO3,HBr. until spots appear. Any secondary spot in the chromatogram
Storage. Store protected from light. spot in the chromatogram obtained with the reference solution.
Other tests. Comply with the tests stated under Eye Drops.
Homatropine Eye Drops Test solution (a). Add 1 ml of a 2 per cent w/v solution of
atropine sulphate RS (internal standard) in methanol (solution
Homatropine Hydrobromide Eye Drops
A) and 1 ml of dilute ammonia solution to a volume of the eye
Homatropine Eye Drops are a sterile solution of Homatropine drops containing about 20 mg of Homatropine Hydrobromide,
Hydrobromide in Purified Water. diluted if necessary to 5.0 ml with water. Extract with two
570
IP 2007 HYALURONIDASE
quantities, each of 5 ml, of chloroform, shake the combined Tests

extracts with 1 g of anhydrous sodium sulphate, filter and
evaporate the filtrate to dryness. Dissolve the residue in 10.0 Appearance of solution. A 1.0 per cent w/v solution in carbon
ml of dichloromethane. To 1.0 ml of this solution add 0.2 ml of dioxide-free water is clear (2.4.1), and not more than faintly
a mixture of 4 volumes of N,O-bis (trimethylsilyl)acetamide yellow.
and 1 volume of trimethylchlorosilane, mix and allow to pH (2.4.24). 4.5 to 7.5, determined in a 0.3 per cent w/v solution
stand for 30 minutes. in carbon dioxide-free water.
Test solution (b). Prepare in the same manner as test solution Light absorption. Dissolve a quantity containing 1500 Units
(a) but omitting the addition of solution A. in sufficient carbon dioxide-free water to produce 5.0 ml and
measure the absorbance of the resulting solution at about
Reference solution. Add 1 ml of solution A and 1 ml of dilute
260 nm and 280 nm; absorbance at about 260 nm, not more
ammonia solution to 5.0 ml of a 0.4 per cent w/v solution of
than 0.42 and at about 280 nm, not more than 0.60 (2.4.7).
homatropine hydrobromide RS and complete the procedure
described under test solution (a) beginning at the words Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit
“Extract with two quantities, each of 5 ml, of chloroform....”. per unit of hyaluronidase.
Chromatographic system Sterility (2.2.11). Complies with the test for sterility.
– a glass column 1.5 m x 4 mm, packed with acid-washed, Assay. The potency of hyaluronidase is determined by
silanised diatomaceous support (80 to 100 mesh) coated comparing its effects against those of the Standard
with 3 per cent w/w of phenyl methyl silicone fluid Preparation.
(50 per cent phenyl) (such as OV-17),
– temperature: Standard Preparation
column. 220°, The Standard Preparation is the 1st International Standard for
inlet port and detector. 280°, Hyaluronidase, bovine, established in 1955, consisting of dried
– flow rate. 30 ml per minute of the carrier gas. bovine testicular hyaluronidase diluted with lactose (supplied
Calculate the content of C16H21NO3,HBr in the eye drops. in ampoules containing 10 tablets of 20 mg each; each tablet
contains approximately 200 Units).
Test solution. Dissolve a suitable quantity of the preparation
Hyaluronidase under examination by adding cold diluent for hyaluronidase
solutions. Dilute the solution with cold diluent for
Hyaluronidase is a material containing enzymes, which hyaluronidase solutions so that the absorbances of the
depolymerise the mucopolysaccharide, hyaluronic acid. It may dilutions being assayed will fall on the upper linear part of the
be prepared from the testes and semen of mammals and purified reference curve prepared as follows.
by fractional precipitation so as to remove inert material and
to which hydrolysed gelatin or a suitable non-protein To each of 12 test-tubes (100 mm x 16 mm) add 0.50 ml of
stabilising agent may be added. The product is freeze-dried in hyaluronate solution and, respectively and in duplicate, 0.5,
single dose containers, which are sealed so as to exclude 0.4, 0.3, 0.2, 0.1 and 0.0 ml of diluent for hyaluronidase
micro-organisms. solutions. If quantities of the solution of the standard solution
other than those indicated below are used, change the
Hyaluronidase contains not less than 300 Units per mg, quantities of diluent for hyaluronidase solutions accordingly.
calculated on the dried basis. It may contain a suitable stabilizer. At intervals of 30 seconds add to the tubes 0.0, 0.1, 0.2, 0.3, 0.4
Description. A white or yellowish-white, fluffy powder. and 0.5 ml of the solution of the standard solution, respectively
and in duplicate, making the final volume in each tube 1.0 ml,
Identification mixing the contents by shaking gently and placing each tube
in a water-bath maintained at 37.0° ± 0.2°. After exactly
A. A solution containing the equivalent of 100 Units in 1 ml of
30 minutes, remove each tube in order from the water-bath at
saline solution depolymerises an equal volume of a 1 per cent
intervals of 30 seconds and immediately add 4.0 ml of serum
w/v solution of sodium hyaluronate at 20° in 1 minute as
solution. Shake and allow to stand at room temperature for 30
shown by a pronounced decrease in viscosity. This action is
minutes. Shake again and measure the absorbance at about
destroyed by heating the initial solution at 100° for 30 minutes.
640 nm (2.4.7). Repeat the operation using 0.50 ml of phosphate-
B. A solution containing the equivalent of 1 Unit in 0.2 ml of buffered saline in place of the hyaluronate solution and make
saline solution when injected intracutaneously into any necessary corrections. Prepare a reference curve by
experimental animals together with a suitable indicator shows plotting the mean of the corrected absorbance for each level
a spreading activity when compared with a control solution. against the potency.
571
HYALURONIDASE INJECTION IP 2007
Standard solution. Dissolve one tablet of the Standard shown by a pronounced decrease in viscosity. This action is
Preparation, accurately weighed, in sufficient cold diluent for destroyed by heating the initial solution at 100° for 30 minutes.
hyaluronidase solutions to give a solution of known B. A solution containing the equivalent of 1 Unit in 0.2 ml of
concentration containing about 1.5 Units per ml. This solution saline solution when injected intracutaneously into
should be prepared immediately before use. experimental animals together with a suitable indicator shows
To each of 6 test-tubes (100 mm x 16 mm) add 0.50 ml of a spreading activity when compared with a control solution.
hyaluronate solution and sufficient diluent for hyaluronidase
solutions so that the final volume in each tube after the addition Tests
of the solution of the preparation being examined is 1.0 ml. At
intervals of 30 seconds add to each tube sufficient of the
solution of the preparation being examined so that the tubes
contain about 0.3, 0.5 and 0.7 Units, respectively and in Appearance of solution. A 1.0 per cent w/v solution in carbon
duplicate, shaking each tube gently and continuing as dioxide-free water is clear (2.4.1), and not more than faintly
described under test solution, beginning at the words “placing yellow.
each tube in a water-bath...” Bacterial endotoxins (2.2.3). Not more than 0.2 Endotoxin Unit
Storage. Store protected from moisture at a temperature not per unit of hyaluronidase.
exceeding 15°. Assay. The potency of hyaluronidase is determined by
Labelling. The label states (1) the total number of Units in the comparing its effects against those of the Standard
container; (2) the name of any added stabilising agent; (3) Preparation.
that the preparation is not intended for intravenous injection.
Standard Preparation
The Standard Preparation is the 1st International Standard for
Hyaluronidase, bovine, established in 1955, consisting of dried
Hyaluronidase Injection bovine testicular hyaluronidase diluted with lactose (supplied
in ampoules containing 10 tablets of 20 mg each; each tablet
Hyaluronidase Injection is a sterile material consisting of contains approximately 200 Units).
Hyaluronidase with or without excipients. It is filled in a sealed
Test solution. Dissolve the contents of a container by adding
container.
cold diluent for hyaluronidase solutions. Dilute the solution
The injection is constituted by dissolving the contents of the with cold diluent for hyaluronidase solutions so that the
sealed container in the requisite amount of sterile Water for absorbances of the dilutions being assayed will fall on the
Injections, immediately before use. upper linear part of the reference curve prepared as follows.
The constituted solution complies with the requirements for To each of 12 test-tubes (100 mm x 16 mm) add 0.50 ml of
Clarity of solution and Particulate matter stated under hyaluronate solution and, respectively and in duplicate, 0.5,
Parenteral Preparations (Injections). 0.4, 0.3, 0.2, 0.1 and 0.0 ml of diluent for hyaluronidase
Storage. The constituted solution should be used immediately solutions. If quantities of the solution of the standard solution
after preparation but, in any case, within the period other than those indicated below are used, change the
recommended by the manufacturer. quantities of diluent for hyaluronidase solutions accordingly.
At intervals of 30 seconds add to the tubes 0.0, 0.1, 0.2, 0.3, 0.4
Hyaluronidase Injection contains not less than 90.0 per cent and 0.5 ml of the solution of the standard solution, respectively
and not more than 115.0 per cent of the stated number of Units and in duplicate, making the final volume in each tube 1.0 ml,
of hyaluronidase activity. mixing the contents by shaking gently and placing each tube
Description. A white or yellowish-white powder. in a water-bath maintained at 37.0° ± 0.2°. After exactly
30 minutes, remove each tube in order from the water-bath at
intervals of 30 seconds and immediately add 4.0 ml of serum
solution. Shake and allow to stand at room temperature for 30
minutes. Shake again and measure the absorbance at about
Identification 640 nm (....). Repeat the operation using 0.50 ml of phosphate-
buffered saline in place of the hyaluronate solution and make
A. A solution containing the equivalent of 100 Units in 1 ml of any necessary corrections. Prepare a reference curve by
saline solution depolymerises an equal volume of a 1 per cent plotting the mean of the corrected absorbance for each level
w/v solution of sodium hyaluronate at 20° in 1 minute as against the potency.
572
IP 2007 HYDRALAZINE HYDROCHLORIDE
Standard solution. Dissolve one tablet of the Standard B. When examined in the range 230 nm to 360 nm (2.4.7), a
Preparation, accurately weighed, in sufficient cold diluent for 0.001 per cent w/v solution shows absorption maxima at about
hyaluronidase solutions to give a solution of known 240 nm, 260 nm, 305 nm and 315 nm and their absorbances are
concentration containing about 1.5 Units per ml. This solution about 0.55, 0.55, 0.27 and 0.22 respectively.
should be prepared immediately before use. C. Dissolve 0.5 g in a mixture of 8 ml of 2 M hydrochloric acid
To each of 6 test-tubes (100 mm x 16 mm) add 0.50 ml of and 100 ml of water. Add 2 ml of sodium nitrite solution, allow
hyaluronate solution and sufficient diluent for hyaluronidase to stand for 10 minutes and filter. The precipitate, after washing
solutions so that the final volume in each tube after the addition with water and drying at 105°, melts at 209° to 212° (2.4.21).
of the solution of the preparation being examined is 1.0 ml. At
D. To a solution of about 10 mg in 2 ml of water add 2 ml of a
intervals of 30 seconds add to each tube sufficient of the
2 per cent w/v solution of 2-nitrobenzaldehyde in ethanol
solution of the preparation being examined so that the tubes
(95 per cent); an orange precipitate is obtained.
contain about 0.3, 0.5 and 0.7 Units, respectively and in
duplicate, shaking each tube gently and continuing as E. A 5 per cent w/v solution gives the reactions of chlorides
described under test solution, beginning at the words “placing (2.3.1).
each tube in a water-bath...”
Tests
exceeding 15°. Appearance of solution. A 0.4 per cent w/v solution is clear
Labelling. The label states (1) the total number of Units (2.4.1), and not more intensely coloured than reference solution
contained in it; (2) the nature of any added stabilising agent; GYS6 (2.4.1).
(4) that the injection should be used immediately after pH (2.4.24). 3.5 to 4.2, determined in a 2.0 per cent w/v solution.
preparation; (5) that the preparation is not intended for
intravenous injection.
(2.4.14).
Test solution (a). Dissolve 25 mg of the substance under
examination in sufficient quantity of the mobile phase to make
50 ml.
Hydralazine Hydrochloride
the mobile phase.
H Reference solution (a). Dilute 5 ml of test solution (b) to 25 ml
N
NH2 , HCl with the mobile phase.
N Reference solution (b). Dissolve 25 mg of phthalazine in
N sufficient quantity of the mobile phase to make 50 ml and
C8H8N4,HCl Mol. Wt. 196.6 dilute 4 ml of this solution to 100 ml with the mobile phase.
Hydralazine Hydrobromide is phthalazin-1-ylhydrazine Reference solution (c). Mix 4 ml of test solution (a) and 10 ml
hydrochloride. of reference solution (b) and dilute to 100 ml with the mobile
phase.
Hydralazine Hydrochloride contains not less 98.5 per cent
and not more than 101.0 per cent of C8H8N4,HCl, calculated on The solutions should be used within 8 hours of preparation.
the dried basis. Chromatographic system
Description. A white or almost white, crystalline powder. – a stainless steel column 25 cm x 4.6 mm, packed with
porous spherical particles of finely-divided silica gel
Identification chemically bonded to nitrile groups (10 µm),
Test A may be omitted if tests B, C, D and E are carried out. and 78 volumes of a solution containing 1.44 g of sodium
Tests B, C and D may be omitted if tests A and E are carried dodecyl sulphate and 0.75 g of tetrabutylammonium
out. bromide per litre adjusted to pH 3.0 with 0.05 M
A. Determine by infrared absorption spectrophotometry (2.4.6). sulphuric acid,
Compare the spectrum with that obtained with hydralazine – flow rate. 1 ml per minute,
hydrochloride RS or with the reference spectrum of – spectrophotometer set at 230 nm,
hydralazine hydrochloride. – a 20 µl loop injector.
573
HYDRALAZINE INJECTION IP 2007
Inject test solution (b) and adjust the sensitivity of the detector with the limit test for heavy metals, Method D (20 ppm). Use
so that the height of the principal peak in the chromatogram is lead standard solution (2 ppm Pb) to prepare the standard
not less than 70 per cent of the full scale of the recorder. When Sulphated ash (2.3.18). Not more than 0.1 per cent, determined
the chromatograms are recorded in the prescribed conditions, on 2.0 g.
the retention time of hydralazine is about 10 to 12 minutes. If
necessary, adjust the concentration of acetonitrile in the mobile Loss on drying (2.4.19). Not more than 0.5 per cent, determined
phase. on 1.0 g by drying over phosphorus pentoxide at a pressure
Inject test solution (a) and continue the chromatography for 3
times the retention time of hydralazine. Inject reference solution Assay. Weigh accurately about 0.1 g and dissolve in a mixture
(a). The area of any secondary peak in the chromatogram of 25 ml of water and 35 ml of hydrochloric acid. Titrate with
obtained with test solution (a) is not greater than the area of 0.05 M potassium iodate, determining the end-point
the peak in the chromatogram obtained with reference solution potentiometrically (2.4.25) and using a calomel reference
(a). electrode and a platinum indicator electrode.
The test is not valid unless (a) the chromatogram obtained 1 ml of 0.05 M potassium iodate is equivalent to 0.009832 g of
with reference solution (c) shows two principal peaks and the C8H8N4,HCl.
resolution between the peaks is not less than 2.5 and (b) the
solution (a) has a signal-to-noise ratio of at least 3. Hydralazine Injection
Hydrazine. Determine by thin-layer chromatography (2.4.17), Hydralazine Hydrochloride Injection
coating the plate with silica gel G.
Hydralazine Injection is a sterile material consisting of
Mobile phase. A mixture of 90 volumes of toluene and 10 Hydralazine Hydrochloride with or without auxiliary
volumes of ethanol (95 per cent). substances. It is filled in a sealed container.
Test solution. Dissolve 0.12 g of the substance under The injection is constituted by dissolving the contents of the
examination in 4 ml of water and 4 ml of a 15 per cent w/v sealed container in the requisite amount of sterile Water for
solution of salicylaldehyde in methanol and 0.2 ml of Injections, immediately before use. For intravenous infusion,
hydrochloric acid. Mix and let it stand at a temperature not the injection should be diluted with an appropriate volume of
exceeding 25° for 2 to 4 hours to allow complete sedimentation a suitable diluent.
of the precipitate. Add 4 ml of toluene, shake vigorously and
centrifuge. Transfer the supernatant liquid to a 100-ml The constituted solution complies with the requirements for
separating funnel, separate the toluene layer and shake Clarity of solution and Particulate matter stated under
vigorously, each time for 3 minutes, with two quantities, each Parenteral Preparations (Injections).
of 20 ml, of a 20 per cent w/v solution of sodium metabisulphite Storage. The constituted solution should be used immediately
and with two quantities, each of 50 ml, of water. Separate the after preparation but, in any case, within the period
toluene layer and use it as the test solution. recommended by the manufacturer.
Reference solution. Prepare at the same time and in the same Hydralazine Injection contains not less than 98.0 per cent and
manner as described for the test solution using 1 ml of a not more than 114.0 per cent of the stated amount of
solution prepared by dissolving 12 mg of hydrazine sulphate hydralazine hydrochloride, C8H8N4,HCl.
in sufficient quantity of 2 M hydrochloric acid to make 100 ml
and diluting 1 ml of this solution to 100 ml with the same Description. A white or almost white powder; very
solvent and 3 ml of water. hygroscopic.
Apply to the plate 20 µl of each solution. Allow the mobile The contents of the sealed container comply with the
phase to rise 10 cm. Dry the plate in air and examine in ultraviolet requirements stated under Parenteral Preparations
light at 365 nm. Any spot in the chromatogram obtained with (Powders for Injection) and with the following requirements.
the test solution showing a yellow fluorescence is not more Identification
reference solution. A. Determine by infrared absorption spectrophotometry (2.4.6).
Heavy metals (2.3.13). Moisten the residue obtained in the Compare the spectrum with that obtained with hydralazine
test for Sulphated ash with 2 ml of hydrochloric acid, hydrochloride RS or with the reference spectrum of
evaporate to dryness and dissolve the residue in sufficient hydralazine hydrochloride.
water to produce 20 ml. 12 ml of the resulting solution complies B. Give the reactions of chlorides (2.3.1).
574
IP 2007 DILUTE HYDROCHLORIC ACID
Tests Description. A clear, colourless, fuming liquid; odour, pungent.
pH (2.4.24). 3.5 to 4.2, determined in a 2.0 per cent w/v solution. Identification
Appearance of solution. A 2.0 per cent w/v solution is not A. When added to potassium permanganate, chlorine is
more opalescent than opalescence standard OS2 (2.4.1). A evolved.
2.0 per cent w/v solution in 0.01 M hydrochloric acid is not
more intensely coloured than reference solution GYS6 (2.4.1). B. Gives the reactions of chlorides (2.3.1).
Hydrazine. Determine by thin-layer chromatography (2.4.17), Tests
using a silica gel 60-precoated plate.
Arsenic (2.3.10). Mix 10.0 g with 40 ml of water and 1 ml of
Mobile phase. The upper layer obtained by shaking together
stannous chloride solution AsT; the resulting solution complies
80 volumes of hexane, 20 volumes of strong ammonia solution
with the limit test for arsenic (1 ppm).
and 20 volumes of ethyl acetate.
Heavy metals (2.3.13). Evaporate 3.5 ml to dryness on a water-
Test solution. Dissolve the contents of a container in sufficient
bath, add 2 ml of dilute acetic acid to the residue and add
0.1 M methanolic hydrochloric acid to produce a solution
water to make 25 ml. The resulting solution complies with the
containing 0.5 per cent w/v of Hydralazine Hydrochloride. To
limit test for heavy metals, Method A (5 ppm).
2.0 ml add 1.0 ml of a 2 per cent w/v solution of salicylaldehyde
in methanol and 0.1 ml of hydrochloric acid, centrifuge and Bromide and iodide. Dilute 5 ml with 10 ml of water, add 1 ml of
decant the supernatant liquid. chloroform and, dropwise with constant shaking, chlorinated
lime solution; the chloroform layer does not become brown
Reference solution. Prepare in the same manner, but using 2.0
or violet.
ml of a 0.00025 per cent w/v solution of hydrazine sulphate in
0.1 M methanolic hydrochloric acid in place of the solution Free chlorine. To 60 ml add 50 ml of carbon dioxide-free
of the substance under examination. water, 1 ml of a 10 per cent w/v solution of potassium iodide
and 0.5 ml of starch solution and allow to stand in the dark for
2 minutes. Any blue colour produced disappears on the
dry the plate in air and spray with dimethylaminobenzaldehyde
addition of 0.2 ml of 0.01 M sodium thiosulphate.
reagent. In the chromatogram obtained with the test solution,
any spot corresponding to hydrazine is not more intense than Sulphite. Dilute 1 ml with 10 ml of water and add 0.25 ml of
the spot in the chromatogram obtained with the reference barium chloride solution and 0.5 ml of 0.001 M iodine; the
solution. colour of the iodine is not completely discharged.
Assay. Determine the weight of the contents of 10 containers. Sulphates (2.3.17). Mix 6.5 ml with 10 mg of sodium
Dissolve 0.1 g of the mixed contents of the 10 containers in a bicarbonate, evaporate to dryness on a water-bath and
mixture of 25 ml of water and 35 ml of hydrochloric acid. dissolve the residue in 15 ml of distilled water. The resulting
Titrate with 0.05 M potassium iodate, determining the end- solution complies with the limit test for sulphates (20 ppm).
point potentiometrically (2.4.25) and using a calomel reference Residue on evaporation. Not more than 0.01 per cent,
electrode and a platinum indicator electrode. determined on 100 g.
1 ml of 0.05 M potassium iodate is equivalent to 0.009832 g of Assay. Weigh accurately about 2.0 g, add 30 ml of water, mix
C8H8N4,HCl. and titrate with 1 M sodium hydroxide using methyl red
Storage. Store protected from light at a temperature not solution as indicator.
exceeding 30°. 1 ml of 1 M sodium hydroxide is equivalent to 0.03646 g of
Labelling. The label states that solutions containing glucose HCl.
should not be used in the preparation of an intravenous Storage. Store in stoppered containers of glass or any other
infusion. inert material at a temperature not exceeding 30°.
Hydrochloric Acid Dilute Hydrochloric Acid

Concentrated Hydrochloric Acid Dilute Hydrochloric Acid is prepared by mixing 274 g of
HCl Mol. Wt. 36.5 Hydrochloric Acid and 726 g of Purified Water.
Hydrochloric Acid contains not less than 35.0 per cent w/w Dilute Hydrochloric Acid contains not less than 9.5 per cent
and not more than 38.0 percent w/w of HCl. and not more than 10.5 per cent w/w of HCl.
575
HYDROCHLOROTHIAZIDE IP 2007
Description. A clear, colourless liquid. Hydrochlorothiazide contains not less than 98.0 per cent and
not more than 102.0 per cent of C7H8ClN3O4S2, calculated on
Identification the dried basis.
A. When added to potassium permanganate, chlorine is Description. A white or almost white, crystalline powder;
evolved. odourless.
B. Gives the reactions of chlorides (2.3.1). Identification
B and D may be omitted if tests A and C are carried out.
stannous chloride AsT; the resulting solution complies with A. Determine by infrared absorption spectrophotometry (2.4.6).
the limit test for arsenic (0.5 ppm). Compare the spectrum with that obtained with
hydrochlorothiazide RS or with the reference spectrum of
Heavy metals (2.3.13). Dissolve the residue obtained in the
hydrochlorothiazide.
test for Residue on evaporation in 1 ml of 2 M hydrochloric
acid, dilute to 25 ml with water; to 2.5 ml of the resulting B. When examined in the range 230 nm to 300 nm (2.4.7), a
solution add 2 ml of dilute acetic acid and add water to make 0.001 per cent w/v solution in 0.01 M sodium hydroxide shows
25 ml. The resulting solution complies with the limit test for an absorption maximum only at about 273 nm; absorbance at
heavy metals, Method A (2 ppm). about 273 nm, 0.5 to 0.54. When examined in the range 300 nm
to 360 nm, a 0.005 per cent w/v solution in 0.01 M sodium
Free chlorine. To 60 ml add 50 ml of carbon dioxide-free
hydroxide shows an absorption maximum at 323 nm;
water, 1 ml of a 10 per cent w/v solution of potassium iodide
absorbance at about 323 nm, 0.45 to 0.48.
and 0.5 ml of starch solution and allow to stand in the dark for
2 minutes. Any blue colour produced disappears on the C. Determine by thin-layer chromatography (2.4.17), coating
addition of 0.2 ml of 0.01 M sodium thiosulphate. the plate with silica gel GF254.
Residue on evaporation. Not more than 0.01 per cent, determined Mobile phase. Ethyl acetate.
on 100 g. Test solution. Dissolve 0.5 g of the substance under
Sulphates (2.3.17). Mix 26 ml with 10 mg of sodium examination in 100 ml of acetone.
bicarbonate, evaporate to dryness on a water-bath and Reference solution. A 0.5 per cent w/v solution of
dissolve the residue in 15 ml of distilled water. The resulting hydrochlorothiazide RS in acetone.
solution complies with the limit test for sulphates (5 ppm).
Assay. Weigh accurately about 6.0 g, add 30 ml of water, mix dry the plate in a current of air and examine in ultraviolet light
and titrate with 1 M sodium hydroxide using methyl red at 254 nm. The principal spot in the chromatogram obtained
solution as indicator. with the test solution corresponds to that in the chromatogram
1 ml of 1 M sodium hydroxide is equivalent to 0.03646 g of obtained with the reference solution.
HCl. D. Heat gently about 1 mg with 2 ml of a freshly prepared 0.05
Storage. Store in stoppered containers of glass or any other per cent w/v solution of chromotropic acid sodium salt in a
inert material at a temperature not exceeding 30°. cooled mixture of 7 volumes of water and 13 volumes of
sulphuric acid; a violet colour develops.
Tests
Hydrochlorothiazide Acidity or alkalinity. Shake 0.5 g of the powdered substance
under examination with 25 ml of water for 2 minutes and filter.
O O O O To 10 ml of the filtrate add 0.2 ml of 0.01 M sodium hydroxide
S S and 0.15 ml of methyl red solution. The solution is yellow and
H2N NH
not more than 0.4 ml of 0.01 M hydrochloric acid is required
Cl N to change the colour of the solution to red.
H
C7H8ClN3O4S2 Mol. Wt. 297.7 (2.4.17), coating the plate with silica gel G.
Hydrochlorothiazide is 6-chloro-3,4-dihydro-2H-1,2,4- Mobile phase. A mixture of 85 volumes of ethyl acetate and 15
benzothiadiazine-7-sulphonamide 1,1-dioxide. volumes of 2-propanol.
576
IP 2007 HYDROCHLOROTHIAZIDE TABLETS
Test solution. Dissolve 0.2 g of the substance under Reference solution. A 0.1 per cent w/v solution of
examination in 10 ml of acetone. hydrochlorothiazide RS in acetone.
Reference solution. A 0.01 per cent w/v solution of the Apply to the plate 5 µl of each solution. After development,
substance under examination in acetone. dry the plate in a current of air, examine in ultraviolet light at
254 nm and then spray with ethanolic sulphuric acid (20 per
cent), heat at 105° for 30 minutes and immediately expose to
dry the plate in a current of air until the odour of the solvent is
nitrous fumes in a closed tank for 15 minutes (the nitrous
no longer detectable and spray with ethanolic sulphuric acid
fumes may be generated by adding 7 M sulphuric acid
(10 per cent); use about 10 ml for a plate (200 mm x 200 mm)
dropwise to a solution containing 10 per cent w/v of sodium
and spray in small portions, allowing the solvent to evaporate
nitrite and 3 per cent w/v of potassium iodide). Place the
each time and avoiding excessive wetting. Heat to 105° for 30
plate in a current of warm air for 15 minutes and spray with a
minutes and immediately place above, but not in, 10 ml of a
0.5 per cent w/v solution of N-(1-naphthyl)ethylenediamine
saturated solution of sodium nitrite in a glass tank. Carefully
dihydrochloride in ethanol (95 per cent). Examine the plate
add 0.5 ml of sulphuric acid to the sodium nitrite solution and
again. By each method of visualisation the principal spot in
allow to stand in the closed tank for 15 minutes. Remove the
the chromatogram obtained with the test solution corresponds
plate, heat it in a ventilated oven at 40° for 15 minutes, spray
to that in the chromatogram obtained with the reference
with three quantities, each of 5 ml, of a freshly prepared 0.5 per
solution.
cent w/v solution of N-(1-naphthyl)ethylenediamine
dihydrochloride in ethanol (95 per cent) and examine by
transmitted light. Any secondary spot in the chromatogram
Tests
obtained with the test solution is not more intense than the Related substances. Determine by thin-layer chromatography
spot in the chromatogram obtained with the reference solution. (2.4.17), coating the plate with silica gel G.
Chlorides (2.3.12). Dissolve 1.0 g in 25 ml of acetone and Mobile phase. A mixture of 85 volumes of ethyl acetate and 15
dilute to 30 ml with water. 7.5 ml of the resulting solution volumes of 2-propanol.
complies with the limit test for chlorides (250 ppm).
Sulphated ash (2.3.18). Not more than 0.1 per cent. containing 50 mg of Hydrochlorothiazide with 50 ml of acetone,
Loss on drying (2.4.19). Not more than 0.5 per cent, determined filter, evaporate the filtrate to dryness and dissolve the residue
on 1.0 g by drying in an oven at 105°. in 10 ml of acetone.
Assay. Weigh accurately about 0.12 g and dissolve in 50 ml of Reference solution. Dilute 1 volume of the test solution to 100
anhydrous pyridine. Titrate with 0.1 M tetrabutylammonium volumes with acetone.
dry the plate in a current of air until the odour of the solvent is
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to no longer detectable and spray with ethanolic sulphuric acid
0.01488 g of C7H8ClN3O4S2. (10 per cent); use about 10 ml for a plate (200 mm x 200 mm)
and spray in small portions, allowing the solvent to evaporate
each time and avoiding excessive wetting. Heat to 105° for 30
minutes and immediately place above, but not in, 10 ml of a
Hydrochlorothiazide Tablets saturated solution of sodium nitrite in a glass tank. Carefully
add 0.5 ml of sulphuric acid to the sodium nitrite solution and
Hydrochlorothiazide Tablets contain not less than 92.5 per allow to stand in the closed tank for 15 minutes. Remove the
cent and not more than 107.5 per cent of the stated amount of plate, heat it in a ventilated oven at 40° for 15 minutes, spray
hydrochlorothiazide, C7H8ClN3O4S2. with three quantities, each of 5 ml, of a freshly prepared 0.5 per
cent w/v solution of N-(1-naphthyl)ethylenediamine
Identification dihydrochloride in ethanol (95 per cent) and examine by
Determine by thin-layer chromatography (2.4.17), coating the transmitted light. Any secondary spot in the chromatogram
plate with silica gel GF254. obtained with the test solution is not more intense than the
Mobile phase. Ethyl acetate.
containing 10 mg of Hydrochlorothiazide with 10 ml of acetone Apparatus. No 1
and filter. Medium. 900 ml of 0.1 M hydrochloric acid
577
HYDROCORTISONE IP 2007
Speed and time. 100 rpm and 45 minutes. B. Determine by thin-layer chromatography (2.4.17), coating
Withdraw a suitable volume of the solution and filter promptly the plate with silica gel GF254.
through a membrane filter disc with an average pore diameter Mobile phase (a). A mixture of 77 volumes of
not greater than 1.0 mm. Reject the first few ml of the filtrate dichloromethane, 15 volumes of ether, 8 volumes of methanol
and dilute a suitable volume of the filtrate with 0.1 M and 1.2 volumes of water.
solution at the maximum at about 272 nm (2.4.7). Calculate the Mobile phase (b). A mixture of 80 volumes of ether, 15 volumes
content of C7H8ClN3O4S2 taking 644 as the specific absorbance of toluene and 5 volumes of 1-butanol saturated with water.
at 272 nm. Solvent mixture. A mixture of 90 volumes of chloroform and
D. Not less than 60 per cent of the stated amount of 10 volumes of methanol.
C7H8ClN3O4S2. Test solution. Dissolve 0.25 g of the substance under
Other tests. Comply with the tests stated under Tablets. examination in 100 ml with solvent mixture.
Assay. Weigh and powder 20 tablets. Weigh accurately a Reference solution. A 0.25 per cent w/v solution of
quantity of the powder containing about 20 mg of hydrocortisone RS in the same solvent mixture.
Hydrochlorothiazide, add 50 ml of 0.1 M sodium hydroxide, Apply to the plate 2 µl of each solution. Develop the
shake for 20 minutes and dilute to 100.0 ml with 0.1 M sodium chromatograms successively with each mobile phase. After
hydroxide. Mix, filter, dilute 5.0 ml of the filtrate to 100.0 ml both developments, dry the plate in air and examine in
with water and measure the absorbance of the resulting ultraviolet light at 254 nm. The principal spot in the
solution at the maximum at about 273 nm (2.4.7). Calculate the chromatogram obtained with the test solution corresponds to
content of C7H8ClN3O4S2 taking 520 as the specific absorbance that in the chromatogram obtained with the reference solution.
at 273 nm. Spray the plate with ethanolic sulphuric acid (20 per cent),
heat at 120° for 10 minutes or until spots appear and allow to
cool. Examine the chromatograms in daylight and in ultraviolet
Hydrocortisone light at 365 nm. The principal spot in the chromatogram
obtained with the test solution is similar in colour in daylight,
Cortisol fluorescence in ultraviolet light at 365 nm, position and size to
the principal spot in the chromatogram obtained with the
O reference solution.
H3C OH
HO OH C. To 2 ml of a 0.1 per cent w/v solution in ethanol (95 per
H3C H cent) add 2 ml of sulphuric acid; an intense yellow colour is
produced with a green fluorescence, which is particularly
H H intense in ultraviolet light at 365 nm. Add the solution to 10 ml
O of water and mix; the fluorescence in ultraviolet light at
365 nm does not disappear.
C21H30O5 Mol. Wt. 362.5
Tests
Hydrocortisone is 11β,17α ,21-trihydroxypregn-4-ene-3,20-
dione. Specific optical rotation (2.4.22). +150° to +156°, determined
Hydrocortisone contains not less than 96.0 per cent and not
more than 104.0 per cent of C21H30O5, calculated on the dried Light absorption (2.4.7). A 0.001 per cent w/v solution in
basis. ethanol (95 per cent) exhibits a maximum at about 240 nm;
absorbance at about 240 nm, between 0.42 and 0.45.
Description. A white to practically white, crystalline powder.
Identification (2.4.14).
Test A may be omitted if tests B and C are carried out. Test C Test solution. Dissolve 25 mg of the substance under
may be omitted if tests A and B are carried out. examination in sufficient tetrahydrofuran to produce 10 ml.
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution (a). Dissolve 2 mg of hydrocortisone RS
Compare the spectrum with that obtained with hydrocortisone and 2 mg of prednisolone RS in the mobile phase and dilute to
RS or with the reference spectrum of hydrocortisone. 100 ml with the mobile phase.
578
IP 2007 HYDROCORTISONE ACETATE
Reference solution (b). Dilute 1 ml of the test solution to Hydrocortisone Acetate

Cortisol Acetate
base deactivated end-capped octadecylsilyl silica gel O
(5 µm), O
– mobile phase: a mixture of 220 ml of tetrahydrofuran H3C O CH3
HO OH
and 700 ml of water, allowed to equilibrate, diluted to
1000 ml with water and mixed again, H3C H
H H
O
Equilibrate the column with the mobile phase for about C23H32O6 Mol. Wt. 404.5
30 minutes.
Hydrocortisone Acetate is 11β,17α-dihydroxy-3,20-
Adjust the sensitivity of the system so that the height of the dioxopregn-4-en-21-yl acetate.
solution (b) is not less than 50 per cent of the full scale of the Hydrocortisone Acetate contains not less than 96.0 per cent
recorder. and not more than 104.0 per cent of C23H32O6, calculated on
the dried basis.
Description. A white or almost white, crystalline powder,
prednisolone, about 14 minutes and hydrocortisone about
odourless.
15.5 minutes. The test is not valid unless the resolution between
the peaks corresponding to prednisolone and hydrocortisone
Identification
is at least 2.2. If necessary, adjust the concentration of
tetrahydrofuran in the mobile phase. Test A may be omitted if tests B, C, and D are carried out.
Inject separately the solvent mixture of the test solution as a Tests C and D may be omitted if tests A and B are carried out.
blank, the test solution and reference solution (b). Continue A. Determine by infrared absorption spectrophotometry (2.4.6).
the chromatography of the test solution for four times the Compare the spectrum with that obtained with hydrocortisone
retention time of the principal peak. In the chromatogram acetate RS or with the reference spectrum of hydrocortisone
obtained with the test solution, the area of any peak other acetate.
than the principal peak, is not greater than half the area of the
solution (b) (0.5 per cent): the sum of the areas of all the peaks
other than the principal peak is not greater than 1.5 times the Mobile phase. Add a mixture of 1.2 volumes of water and 8
area of the principal peak in the chromatogram obtained with volumes of methanol to a mixture of 15 volumes of ether and
reference solution (b) (1.5 per cent). Ignore any peak obtained 77 volumes of dichloromethane.
with the blank and any peak with an area less than 0.05 times Test solution (a). Dissolve 25 mg of the substance under
that of the principal peak in the chromatogram obtained with examination in methanol and dilute to 5 ml with the same
reference solution (b). solvent. Use this solution to prepare test solution (b). Dilute 2
Sulphated ash (2.3.18). Not more than 0.1 per cent. ml of the solution to 10 ml with dichloromethane.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Test solution (b). Transfer 2 ml of the solution obtained in
on 1.0 g by drying in an oven at 105° for 3 hours. preparing test solution (a) to a 15-ml glass tube with a glass or
plastic stopper. Add 10 ml of saturated methanolic potassium
Assay. Weigh accurately about 0.1 g, dissolve in sufficient hydrogen carbonate solution and immediately pass a stream
ethanol to produce 100.0 ml. Dilute 2.0 ml of this solution to of nitrogen through the solution for 5 minutes. Stopper the
100.0 ml with ethanol and mix. Measure the absorbance of the tube. Heat in a water-bath at 45° protected from light for 2½
resulting solution (2.4.7) at the maximum at 241.5 nm. Calculate hours. Allow to cool.
the content of C21H30O5 taking 440 as the specific absorbance
at 241.5 nm. Reference solution (a). Prepare in the same manner as test
solution (a) but using hydrocortisone acetate RS in place of
Storage. Store protected from light. the substance under examination.
579
HYDROCORTISONE EYE OINTMENT IP 2007
Reference solution (b). Prepare in the same manner as test Adjust the sensitivity of the system so that the height of the
solution (b) but using 2 ml of the solution obtained in preparing principal peak in the chromatogram obtained with reference
reference solution (a) in place of the solution obtained in solution (b) is not less than 50 per cent of the full scale of the
preparing test solution (a). recorder.
Apply to the plate 5 µl of each solution. After development, Inject reference solution (a). The retention times are:
dry the plate in air and examine in ultraviolet light at 254 nm. hydrocortisone acetate, about 10 minutes and cortisone
The principal spot in each of the chromatograms obtained acetate about 12 minutes. The test is not valid unless the
with the test solutions is similar to the principal spot in the resolution between the peaks corresponding to hydrocortisone
chromatogram obtained with the corresponding reference acetate and cortisone acetate is at least 4.2. If necessary, adjust
solution. Spray with ethanolic sulphuric acid (20 per cent the concentration of acetonitrile in the mobile phase.
v/v) and heat at 120° for 10 minutes or until the spots appear.
Inject separately the test solution and reference solution (b).
Allow to cool and examine in daylight and in ultraviolet light
Continue the chromatography of the test solution for 2.5 times
at 365 nm. The principal spot in each of the chromatograms
the retention time of the principal peak. In the chromatogram
obtained with the test solutions is similar in position, colour
obtained with the test solution: the area of any peak other
in daylight, fluorescence in ultraviolet light at 365 nm, position
than the principal peak, is not greater than the area of the
and size to that in the chromatogram obtained with the
corresponding reference solution. The principal spots in the
solution (b) (1.0 per cent) and not more than one such peak
chromatograms obtained with test solution (b) and reference
has an area greater than half the area of the peak in the
solution (b) have an Rf value distinctly lower than that of the
principal spots in the chromatograms obtained with test
cent); the sum of the areas of all the peaks other than the
solution (a) and reference solution (a).
principal peak is not greater than 1.5 times the area of the
D. Gives the reaction of acetyl groups (2.3.1). principal peak in the chromatogram obtained with reference
solution (b) (1.5 per cent). Ignore any peak due to the solvent
Tests and any peak with an area less than 0.05 times that of the
Specific optical rotation (2.4.22). +158° to +167°, determined principal peak in the chromatogram obtained with reference
in a 1.0 per cent w/v solution in dioxan. solution (b).
Light absorption (2.4.7). A 0.001 per cent w/v solution in Sulphated ash (2.3.18). Not more than 0.1 per cent.
ethanol (95 per cent) shows an absorption maximum at about Loss on drying (2.4.19). Not more than 0.5 per cent, determined
240 nm; absorbance at about 240 nm, between 0.38 and 0.40. on 2.0 g by drying in an oven at 105° for 3 hours.
(2.4.14).
ethanol to produce 100.0 ml. Dilute 2.0 ml of this solution to
Test solution. Dissolve 25 mg of the substance under 100.0 ml with ethanol and mix. Measure the absorbance of the
examination in sufficient methanol to produce 10 ml. resulting solution (2.4.7) at the maximum at 241.5 nm. Calculate
Reference solution (a). Dissolve 2 mg of hydrocortisone the content of C23H32O6 taking 395 as the specific absorbance
acetate RS and 2 mg of cortisone acetate RS in the mobile at 241.5 nm.
phase and dilute to 100 ml with the mobile phase. Storage. Store protected from light.
Reference solution (b). Dilute 1 ml of the test solution to
Chromatographic system Hydrocortisone Eye Ointment
– a stainless steel column 25 cm x 4.6 mm, packed with Hydrocortisone Acetate Eye Ointment; Cortisol
base deactivated end-capped octadecylsilyl silica gel
Acetate Eye Ointment
(5 µm),
– mobile phase: a mixture of 400 ml of acetonitrile and 550 Hydrocortisone Eye Ointment contains not less than 90.0 per
ml of water, allowed to equilibrate, diluted to cent and not more than 110.0 per cent of the stated amount of
1000 ml with water and mixed again, hydrocortisone acetate, C23H32O6.
– a 20 µl loop injector. Boil 2 g with 20 ml of methanol, shake, cool to 0° for 30 minutes,
Equilibrate the column with the mobile phase for about 30 filter and evaporate the filtrate to dryness. The residue complies
minutes. with the following tests.
580
IP 2007 HYDROCORTISONE ACETATE INJECTION
A. Determine by thin-layer chromatography (2.4.17), coating Chromatographic system

the plate with silica gel G. – a stainless steel column 30 cm x 3.9 mm, packed with
Solvent mixture. A mixture of 90 volumes of acetone and 10 porous silica particles (5 µm),
volumes of formamide. – mobile phase: a mixture of 425 volumes of butyl chloride
saturated with water, 70 volumes of tetrahydrofuran,
Mobile phase. A mixture of 30 volumes of toluene and 10 35 volumes of methanol and 30 volumes of glacial acetic
volumes of chloroform. acid,
Test solution. Dissolve 25 mg of the substance under – flow rate. 1 ml per minute,
examination in 10 ml of the solvent mixture. – spectrophotometer set at 254 nm,
Reference solution (a). Dissolve 25 mg of hydrocortisone RS – a 20 µl loop injector.
in 10 ml of the solvent mixture. Calculate the content of C23H32O6 in the ointment.
Reference solution (b). Mix equal volumes of the test solution Storage. Store protected from light at a temperature not
and reference solution (a). exceeding 30°.
solvent mixture, allow the solvent mixture to ascend to the
top, remove the plate from the tank and allow the solvent to
Hydrocortisone Acetate Injection
was done. Cortisol Acetate Injection
Apply to the plate 2 µl of each solution. Allow the mobile Hydrocortisone Acetate Injection is a sterile suspension of a
phase to rise 12 cm. Dry the plate in a current of warm air, allow very fine powder of Hydrocortisone Acetate in Water for
the solvent to evaporate, heat at 120° for 15 minutes and spray Injections or Sodium Chloride Injection containing suitable
the hot plate with ethanolic sulphuric acid (20 per cent v/v). dispersing agents.
Hydrocortisone Acetate Injection contains not less than 95.0
per cent and not more than 105.0 per cent of the stated amount
of hydrocortisone acetate, C23H32O6.
Identification
compact spot. Filter a volume containing 50 mg of Hydrocortisone Acetate
B. Dissolve about 1 mg in 1 ml of sulphuric acid and mix; an through a sintered-glass filter, wash the residue with four
orange colour is produced with a green fluorescence, which is quantities, each of 5 ml, of water, dissolve in 20 ml of
particularly intense under ultraviolet light at 365 nm. Add the chloroform, wash the chloroform solution with four quantities,
solution to 10 ml of water and mix; the fluorescence under each of 10 ml, of water, discard the washings, filter the
ultraviolet light at 365 nm does not disappear. chloroform solution through a plug of cotton and evaporate
the filtrate to dryness. The residue complies with the following
C. Dissolve 10 mg in 1 ml of methanol, warm and add 1 ml of
tests.
potassium cupri-tartrate solution; an orange-red precipitate
is slowly formed. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with hydrocortisone
Tests acetate RS or with the reference spectrum of hydrocortisone
Other tests. Complies with the tests stated under Eye acetate.
Ointments. B. Determine by thin-layer chromatography (2.4.17), coating
Assay. Determine by liquid chromatography (2.4.14). the plate with silica gel G.
Test solution. Weigh accurately a quantity of the ointment Solvent mixture. A mixture of 90 volumes of acetone and 10
containing about 10 mg of Hydrocortisone Acetate, shake volumes of formamide.
with 20.0 ml of methanol for 30 minutes and centrifuge. To
10.0 ml of the clear, supernatant layer add sufficient methanol Mobile phase. A mixture of 30 volumes of toluene and 10
to produce 50.0 ml. volumes of chloroform.
Reference solution. A solution containing 0.01 per cent w/v of Test solution. Dissolve 25 mg of the substance under
hydrocortisone acetate RS in methanol. examination in 10 ml of the solvent mixture.
581
HYDROCORTISONE HEMISUCCINATE IP 2007
Reference solution (a). Dissolve 25 mg of hydrocortisone Hydrocortisone Hemisuccinate

acetate RS in 10 ml of the solvent mixture.
Hydrocortisone Hydrogen Succinate; Cortisol
Hydrogen Succinate
Place the dry plate in a tank containing a shallow layer of the O

solvent mixture, allow the solvent mixture to ascend to the O
top, remove the plate from the tank and allow the solvent to H3C O COOH
evaporate. Use within 2 hours, with the flow of the mobile HO OH
phase in the direction in which the aforementioned treatment H3C H
was done.
H H
O
the solvent to evaporate, heat at 120° for 15 minutes and spray C25H34O8 Mol. Wt. 462.6
Heat at 120° for a further 10 minutes, allow to cool and examine Hydrocortisone Hemisuccinate is 11β,17α -dihydroxy-3, 20-
in daylight and in ultraviolet light at 365 nm. The principal dioxopregn-4-en-21-yl hydrogen succinate.
spot in the chromatogram obtained with the test solution Hydrocortisone Hemisuccinate contains not less than 98.0
corresponds to that in the chromatogram obtained with per cent and not more than 101.0 per cent of C25H34O8,
reference solution (a). The principal spot in the chromatogram calculated on the dried basis.
compact spot.
hygroscopic.
Other tests. Complies with the tests stated under Parenteral Test A may be omitted if tests B, C and D are carried out. Tests
Preparations (Injections) C and D may be omitted if tests A and B are carried out.
Assay. Determine by liquid chromatography (2.4.14). A. Determine by infrared absorption spectrophotometry (2.4.6).
Test solution. To an accurately measured volume of the injection Dry the substances before use at 105° for 3 hours and examine
containing about 50 mg of Hydrocortisone Acetate add 70 ml them as discs. Compare the spectrum with that obtained with
of methanol, shake to produce a clear solution and dilute to hydrocortisone hemisuccinate RS or with the reference
100.0 ml with methanol. Dilute 10.0 ml of the resulting solution spectrum of hydrocortisone hemisuccinate.
to 20.0 ml with water. B. Determine by thin-layer chromatography (2.4.17), coating
Reference solution. Dissolve 25.0 mg of hydrocortisone
acetate RS in 50 ml of methanol and add sufficient water to Mobile phase. Add a mixture of 1.2 volumes of water and 8
produce 100.0 ml. volumes of methanol to a mixture of 15 volumes of ether and
77 volumes of dichloromethane.
– a stainless steel column 10 cm x 5 mm, packed with Test solution (a). Dissolve 25 mg of the substance under
octadecylsilyl silica gel (5 µm) (such as Spherisorb examination in methanol and dilute to 5 ml with the same
ODS 1), solvent. Use this solution to prepare test solution (b). Dilute 2
– mobile phase: 50 per cent v/v solution of methanol, ml of the solution to10 ml with dichloromethane.
– flow rate. 2 ml per minute, Test solution (b). Transfer 2 ml of the solution obtained in
– spectrophotometer set at 240 nm, preparing test solution (a) to a 15-ml glass tube with a glass or
– a 20 µl loop injector. plastic stopper. Add 10 ml of a 0.8 g per litre solution of sodium
Calculate the content of C23H32O6 in the injection. hydroxide in methanol and immediately pass a stream of
nitrogen through the solution for 5 minutes. Stopper the tube.
Storage.Store protected from light. Heat in a water-bath at 45° protected from light for 30 minutes.
Labelling. The label states (1) that the contents are meant for Allow to cool.
local injection only; (2) that the container should be gently Reference solution (a). Prepare in the same manner as test
shaken before a dose is withdrawn; (3) the names of the solution (a) but using hydrocortisone hemisuccinate RS in
dispersing agents used, if any. place of the substance under examination.
582
IP 2007 HYDROCORTISONE SODIUM SUCCINATE INJECTION
Reference solution (b). Prepare in the same manner as test – mobile phase: a mixture of 330 ml of acetonitrile and 600
solution (b) but using 2 ml of the solution obtained in preparing ml of water and 1.0 ml of phosphoric acid, allowed to
reference solution (a) in place of the solution obtained in equilibrate, diluted to 1000 ml with water and mixed again,
preparing test solution (a). – flow rate. 1 ml per minute,
dry the plate in air and examine in ultraviolet light at 254 nm. – a 20 µl loop injector.
The principal spot in each of the chromatograms obtained Equilibrate the column with the mobile phase for about 30
with the test solutions is similar to the principal spot in the minutes.
chromatogram obtained with the corresponding reference Adjust the sensitivity of the system so that the height of the
solution. Spray with ethanolic sulphuric acid (20 per cent principal peak in the chromatogram obtained with reference
v/v) and heat at 120° for 10 minutes or until the spots appear. solution (b) is not less than 50 per cent of the full scale of the
Allow to cool and examine in daylight and in ultraviolet light recorder.
at 365 nm. The principal spot in each of the chromatograms
obtained with the test solutions is similar in position, colour Inject reference solution (a). The retention times are:
in daylight, fluorescence in ultraviolet light at 365 nm, position dexamethasone, about 12.5 minutes and hydrocortisone
and size to that in the chromatogram obtained with the hemisuccinate about 15 minutes. The test is not valid unless
corresponding reference solution. The principal spots in the the resolution between the peaks corresponding to
chromatograms obtained with test solution (b) and reference dexamethasone and hydrocortisone hemisuccinate is at least
solution (b) have an Rf value distinctly lower than that of the 5.0. If necessary, adjust the concentration of acetonitrile in
principal spots in the chromatograms obtained with test the mobile phase.
solution (a) and reference solution (a). Inject separately the test solution and reference solution (b).
Continue the chromatography for twice the retention time of
C. Add about 2 mg to 2 ml of sulphuric acid; a yellow to
the principal peak. In the chromatogram obtained with the test
brownish red colour develops with a green fluorescence that
solution: the area of any peak other than the principal peak, is
is particularly intense when examined in ultraviolet light at
not greater than half the area of the principal peak in the
365 nm.
D. About 30 mg gives the reaction of esters (2.3.1). cent); the sum of the areas of all the peaks other than the
principal peak is not greater than 0.75 times the area of the
Tests principal peak in the chromatogram obtained with reference
solution (b) (0.75 per cent). Ignore any peak due to the solvent
Appearance of solution. A 2.0 per cent w/v solution in sodium
and any peak with an area less than 0.05 times that of the
bicarbonate solution is clear (2.4.1).
Light absorption (2.4.7). A 0.001 per cent w/v solution in solution (b).
ethanol (95 per cent) shows an absorption maximum at about Sulphated ash (2.3.18). Not more than 0.1 per cent.
240 nm; absorbance at about 240 nm, between 0.34 and 0.36.
Specific optical rotation (2.4.22). +147° to +153°, determined on 1.0 g by drying in an oven at 105°.
Related substances. Determine by liquid chromatography ethanol to produce 100.0 ml. Dilute 2.0 ml of this solution to
(2.4.14). 100.0 ml with ethanol and mix. Measure the absorbance of the
Test solution. Dissolve 25 mg of the substance under resulting solution (2.4.7) at the maximum at about 241.5 nm.
examination in sufficient methanol to produce 10 ml. Calculate the content of C25H34O8 taking 353 as the specific
absorbance at 241.5 nm.
Reference solution (a). Dissolve 2 mg of hydrocortisone
hemisuccinate RS and 2 mg of dexamethasone RS in the mobile Storage. Store protected from light and moisture.
phase and dilute to 100 ml with the mobile phase.
Reference solution (b). Dilute 1 ml of the test solution to100 Hydrocortisone Sodium Succinate
Injection
– a stainless steel column 25 cm x 4.6 mm, packed with Cortisol Sodium Succinate Injection
base deactivated end-capped octadecylsilyl silica gel Hydrocortisone Sodium Succinate Injection is a sterile material
(5 µm), made from Hydrocortisone Hemisuccinate with the aid of a
583
HYDROCORTISONE SODIUM SUCCINATE INJECTION IP 2007
suitable alkali such as Sodium Hydroxide or Sodium Carbonate. Tests

It may contain suitable buffering agents. It is filled in a sealed
container. pH (2.4.24). 6.5 to 8.0, determined in a solution containing the
equivalent of 5.0 per cent w/v of hydrocortisone.
sealed container in the requisite amount of sterile Water for Related substances. Determine by liquid chromatography
Injections, immediately before use. (2.4.14).
The constituted solution complies with the requirements for Solvent mixture. A mixture of equal volumes of acetonitrile
Clarity of solution and Particulate matter stated under and water.
Parenteral Preparations (Injections). Test solution. Dissolve a sufficient quantity of the contents
Storage. The constituted solution should be used immediately of the sealed container in a solvent mixture to produce a
after preparation but, in any case, within the period solution containing the equivalent of 0.25 per cent w/v of
recommended by the manufacturer. hydrocortisone.
Hydrocortisone Sodium Succinate Injection contains not less Reference solution (a). Dilute 2 volumes of the test solution
than 95.0 per cent and not more than 105.0 per cent of the to 100 volumes with solvent mixture.
stated amount of hydrocortisone, C21H30O5. Reference solution (b). Dilute a 0.035 per cent w/v solution of
The contents of the sealed container comply with the hydrocortisone RS in acetonitrile with solvent mixture.
requirements stated under Parenteral Preparations Reference solution (c). Dilute a solution containing 0.04 per
(Powders for Injection) and with the following requirements. cent w/v each of hydrocortisone hemisuccinate RS and
dexamethasone RS with solvent mixture.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). – a stainless steel column 25 cm x 4.6 mm, packed with
Compare the spectrum with that obtained with the reference octadecylsilyl silica gel (5 µm),
spectrum of hydrocortisone hemisuccinate. – mobile phase: a mixture of 330 ml of acetonitrile and 600
ml of water and 1 ml of phosphoric acid, allowed to
equilibrate, diluted to 1000 ml with water and mixed again,
Mobile phase. A freshly prepared mixture of 1 volume of – spectrophotometer set at 254 nm,
anhydrous formic acid, 10 volumes of absolute ethanol and – a 20 µl loop injector.
Equilibrate the column with the mobile phase for about 30
Test solution. A 0.1 per cent w/v solution of the contents of minutes.
the sealed container in a mixture of 1 volume of methanol and
Inject reference solution (c). The test is not valid unless in the
chromatogram obtained the resolution between the peaks
Reference solution (a). A 0.1 per cent w/v solution of corresponding to dexamethasone and hydrocortisone
hydrocortisone sodium succinate RS in the same solvent hemisuccinate is at least 5.0.
mixture.
Inject the test solution. Allow the chromatography to proceed
Reference solution (b). A solution containing 0.1 per cent for twice the retention time of the principal peak. In the
w/v each of hydrocortisone hemisuccinate RS and chromatogram obtained with the test solution the area of any
methylprednisolone hydrogen succinate RS in the same peak corresponding ro hydrocortisone is not greater than the
solvent mixture. area of the peak in the chromatogram obtained with reference
Apply separately to the plate 5 µl of each solution. After solution (b) (7 per cent) and the area of any other secondary
development, dry the plate in air until the solvents have peak is not greater than the area of the peak in the
evaporated, spray with ethanolic sulphuric acid (20 per cent), chromatogram obtained with reference solution (a) (2 per cent).
heat at 120° for 10 minutes, allow to cool and examine in daylight Assay. Determine the weight of the contents of 10 containers.
and in ultraviolet light at 365 nm. By each method of Dissolve the mixed contents of the 10 containers in sufficient
visualization the principal spot in the chromatogram obtained water to produce a solution containing the equivalent of 0.001
with the test solution corresponds to that in the chromatogram per cent w/v of hydrocortisone. Measure the absorbance of
obtained with reference solution (a). The test is not valid unless the resulting solution at the maximum at about 248 nm (2.4.7).
the chromatogram obtained with reference solution (b) shows Calculate the content of C21H30O5 taking 449 as the specific
two spots that may not be completely separated. absorbance at 248 nm.
584
IP 2007 HYDROGEN PEROXIDE SOLUTION (100 VOL)
Storage. Store protected from moisture in single dose 1 ml of 0.02 M potassium permanganate is equivalent to
containers at a temperature not exceeding 30°. 0.001701 g of H2O2 or 0.56 ml of oxygen.
Labelling. The label states (1) the strength in terms of the Storage. Store protected from light in containers resistant to
equivalent amount of hydrocortisone; (2) that the prepared hydrogen peroxide at a temperature not exceeding 30°. If the
solution should be used only if it is clear; (3) that the solution solution does not contain a stabilising agent, it should be
should be used immediately after preparation. stored in a refrigerator (2° to 8°). It should not be stored for
long periods.
Labelling. The label states whether or not the solution
Hydrogen Peroxide Solution (20 Vol) contains a stabilising agent.
H2O2 Mol. Wt. 34.0

Hydrogen Peroxide Solution (6 per cent); Dilute Hydrogen Peroxide Solution (100 Vol)
Hydrogen Peroxide Solution
Hydrogen Peroxide Solution (27 per cent); Strong
Hydrogen Peroxide Solution (20 Vol) is an aqueous solution Hydrogen Peroxide Solution
of hydrogen peroxide. It may contain a suitable stabilising
agent. Hydrogen Peroxide Solution (100 Vol) is an aqueous solution
of hydrogen peroxide. It may contain a suitable stabilising
Hydrogen Peroxide Solution (20 Vol) contains not less than agent.
5.0 per cent w/v and not more than 7.0 per cent w/v of H2O2,
corresponding to about 20 times its volume of available Hydrogen Peroxide Solution (100 Vol) contains not less than
oxygen. 26.0 per cent w/w and not more than 28.0 per cent w/w of H2O2,
corresponding to about 100 times its volume of available
Description. A clear, colourless liquid; odourless. It oxygen.
decomposes in contact with oxidisable organic matter and
with certain metals and also if allowed to become alkaline. Description. A clear, colourless liquid; odourless. It
decomposes vigorously in contact with oxidisable organic
Identification matter and with certain metals and also if allowed to become
alkaline.
A. To 1 ml add 0.2 ml of 1 M sulphuric acid and 0.25 ml of 0.02
M potassium permanganate; the solution becomes colourless Identification
with evolution of gas.
A. To 1 ml add 0.2 ml of 1 M sulphuric acid and 0.25 ml of 0.02
B. Shake 0.05 ml with 2 ml of 1 M sulphuric acid, 2 ml of ether M potassium permanganate; the solution becomes colourless
and 0.05 ml of potassium chromate solution; the ether layer with evolution of gas.
becomes blue.
B. Shake 0.05 ml with 2 ml of 1 M sulphuric acid, 2 ml of ether
Tests and 0.05 ml of potassium chromate solution; the ether layer
becomes blue.
Acidity. To 10 ml add 20 ml of water and 0.25 ml of methyl red
solution. Not less than 0.2 ml and not more than 1.0 ml of Tests
Acidity. Dilute 10 ml with 100 ml of water and add 0.25 ml of
the solution.
methyl red solution. Not less than 0.05 ml and not more than
Organic stabilizers. Shake 20 ml with successive quantities 0.5 ml of 0.1 M sodium hydroxide is required to change the
of 10, 5 and 5 ml of chloroform. Evaporate the combined colour of the solution.
chloroform extracts at a temperature not exceeding 25° at a
Organic stabilisers. Shake 20 ml with successive quantities
pressure of 2 kPa and dry in a desiccator. The residue weighs
of 10, 5 and 5 ml of chloroform. Evaporate the combined
not more than 10 mg (500 ppm).
chloroform extracts at a temperature not exceeding 25° at a
Non-volatile matter. Place 10 ml in a platinum dish and allow pressure of 2 kPa and dry in a desiccator. The residue weighs
to stand until effervescence ceases. Evaporate the solution not more than 10 mg (500 ppm).
on a water-bath and dry the residue at 105°; the residue weighs
Non-volatile matter. Place 10 ml in a platinum dish and allow
not more than 20 mg.
to stand until effervescence ceases. Evaporate the solution
Assay. To 1.0 ml add 20 ml of 1 M sulphuric acid and titrate on a water-bath and dry the residue at 105°; the residue weighs
with 0.02 M potassium permanganate. not more than 20 mg.
585
HYDROXOCOBALAMIN IP 2007
Assay. Dilute about 1.0 g to 100.0 ml with water. To 10.0 ml of Identification

the resulting solution add 20 ml of 1 M sulphuric acid and
titrate with 0.02 M potassium permanganate. A. Measure the absorbance of the solution used in the Assay
at the maxima at about 274 nm, 351 nm and 525 nm (2.4.7);
1 ml of 0.02 M potassium permanganate is equivalent to ratios of the absorbances at about 274 nm and 525 nm to that
0.001701 g of H2O2 or 0.56 ml of oxygen. at about 351 nm, about 0.8 and about 0.3 respectively.
Storage. Store protected from light in containers resistant to
B. Fuse 1 mg of the substance with 50 mg of potassium
hydrogen peroxide at a temperature not exceeding 30°. If the
hydrogen sulphate, cool, break up the mass, add 3 ml of water
solution does not contain a stabilising agent, it should be
and boil until dissolved. Add 0.05 ml of phenolphthalein
stored in a refrigerator (2° to 8°). It should not be stored for
solution and sufficient 5 M sodium hydroxide to produce a
long periods.
faint pink colour. Add 0.5 g of sodium acetate, 0.5 ml of 1 M
Labelling. The label states whether or not the solution acetic acid and 0.5 ml of a 0.2 per cent w/v solution of nitroso
contains a stabilising agent. R salt; a red or orange-red colour is produced immediately.
Add 0.5 ml of hydrochloric acid and boil for 1 minute; the red
colour persists.
Hydroxocobalamin C. Gives the reactions of chlorides or of sulphates, as the case
may be (2.3.1).
H2N Tests
O
O
H2N Related substances. Determine by liquid chromatography
H CH3 CH3 O
(2.4.14).
O H3C
NH2 Use freshly prepared solutions and protect them from bright
H
H2N H C N OH N light.
3 Co
H N Test solution. Dissolve 10 mg of the substance under
O H N
CH3 examination in the mobile phase and dilute to 10 ml with the
CH3 mobile phase.
H2N NH2
H CH3 CH3 H Reference solution (a). Dilute 5 ml of the test solution to 100
N
O ml with the mobile phase.
N CH3
H3C O Reference solution (b). Dilute 1 ml of the test solution to 10 ml
H O
N with the mobile phase. Dilute 1 ml of this solution to 100 ml
O P CH3
O O HO
Reference solution (c). Dissolve 25 mg of the substance under
examination in 10 ml of water, warming if necessary. Allow to
O cool and add 1 ml of a 20 g/l solution of chloramine T and 0.5
HO ml of 0.05 M hydrochloric acid. Dilute to 25 ml with water.
Shake and allow to stand for 5 minutes. Inject immediately.
C62H89CoN13O15P Mol. Wt. 1346.4
Hydroxocobalamin is Coα-[α-(5,6-dimethylbenzimidazolyl)]- octylsilyl silica gel (5 µm),
Coβ-hydroxocobamide. It occurs either as aquocobalamin – mobile phase: a mixture of 19.5 volumes of methanol
chloride (Coα-[α-(5,6-dimethylbenzimidazolyl)]-ICoβ- and 80.5 volumes of a solution containing 15 g per litre
aquocobamide chloride) or as aquocobalamin sulphate. of citric acid and 8.1g of disodium hydrogen phosphate,
Hydroxocobalamin contains not less than 96.0 per cent and – flow rate. 1.5 ml per minute,
not more than 102.0 per cent of aquocobalamin chloride, – spectrophotometer set at 351 nm,
C 62 H 89 CoN 13 O 15P,HCl, or of aquocobalamin sulphate, – a 20 µl loop injector.
C124H178Co2N26O30P2,H2SO4, both calculated on the dried basis. Inject reference solution (c). The test is not valid unless the
Description. Dark red crystals or a crystalline powder; very chromatogram obtained shows three principal peaks and the
hygroscopic. Some decomposition may occur on drying. resolution between each pair of adjacent peaks is at least 3.0.
586
IP 2007 HYDROXOCOBALAMIN INJECTION
Inject reference solution (b). The chromatogram obtained Related substances. Determine by liquid chromatography
shows one principal peak with a signal-to-noise ratio of at (2.4.14).
least 5. Use freshly prepared solutions and protect them from bright
Inject the test solution and reference solution (a). Continue light.
the chromatography for four times the retention time of the Test solution. Dilute the injection with the mobile phase, if
principal peak in the chromatogram obtained with reference necessary, to obtain a solution having a concentration of
solution (a). In the chromatogram obtained with the test 0.1 per cent w/v of hydroxocobalamin.
solution, the sum of the areas of any peaks apart from the
principal peak is not greater than the area of the principal peak Reference solution (a). Dilute 1 ml of the test solution to 20 ml
in the chromatogram obtained with reference solution (a) (5 with the mobile phase.
per cent). Ignore any peak whose area is less than that of the Reference solution (b). Dilute 1 ml of reference solution (a) to
principal peak in the chromatogram obtained with reference 100 ml with the mobile phase.
solution (b).
Reference solution (c). Add 0.2 ml of a freshly prepared 2 per
Loss on drying (2.4.19). 8 to 12 per cent (aquocobalamin cent w/v solution of chloramine T and 0.1 ml of 0.05 M
chloride) and 8 to 16 per cent (aquocobalamin sulphate), hydrochloric acid to a volume of the injection containing the
determined on 1.0 g by drying in an oven at 100° at a pressure equivalent of 5 mg of hydroxocobalamin, dilute to 10 ml with
not exceeding 0.7 kPa. water, shake, allow to stand for 5 minutes and inject
Assay. Protect the solutions from light throughout the Assay. immediately.
Weigh accurately about 25 mg and dissolve in sufficient of a Chromatographic system

solution containing 0.8 per cent v/v of glacial acetic acid – a stainless steel column 25 cm × 4 mm, packed with
and 1.09 per cent w/v of sodium acetate to produce 1000 ml. octylsilyl silica gel (5 µm) (such as Lichrosorb 100 CH 8/
Measure the absorbance of the resulting solution at the 11),
maximum at about 351 nm (2.4.7). Calculate the content of – mobile phase: a mixture of 19.5 volumes of methanol
C62H89CoN13O15P,HCl, or of C124H178Co2N26O30P2,H2SO4, taking and 80.5 volumes of a solution containing 1.5 per cent
190 or 188 respectively, as the specific absorbance at 351 nm. w/v of citric acid and 0.81per cent w/v of disodium
hydrogen orthophosphate,
Storage. Store protected from light and moisture at a – flow rate. 1.5 ml per minute,
temperature not exceeding 30°. – spectrophotometer set at 351 nm,
Labelling. The label states whether the contents are – a 20 µl loop injector.
aquocobalamin chloride or aquocobalamin sulphate. Inject reference solution (c). The test is not valid unless the
chromatogram obtained shows three principal peaks and the
resolution between each pair of adjacent peaks is not less
than 3.0.
Hydroxocobalamin Injection Inject reference solution (b). The chromatogram obtained
shows one principal peak with a signal-to-noise ratio of not
Hydroxocobalamin Injection is a sterile solution of
less than 5.
Hydroxocobalamin in Water for Injections containing
sufficient Acetic Acid, Hydrochloric Acid or Sulphuric acid to Inject the test solution and reference solution (a). In the
adjust the pH to about 4. chromatogram obtained with the test solution the sum of the
areas of any secondary peaks is not greater than twice the
Hydroxocobalamin Injection contains not less than 95.0 per
reference solution (a) (10 per cent). Ignore any peak the area
anhydrous hydroxocobalamin, C62H89CoN13O15P.
of which is less than that of the principal peak in the
Identification cent).
Measure the absorbance at about 351 nm and 361 nm; ratio of Other tests. Complies with the tests stated under Parenteral
the absorbance at about 361 nm to that at about 351 nm, about Preparations (Injections).
0.65 (2.4.7).
Assay. Dilute an accurately measured volume of the injection
Tests containing about 4 mg of anhydrous hydroxocobalamin to
200.0 ml with a solution containing 0.8 per cent v/v of glacial
pH (2.4.24). 3.8 to 5.5. acetic acid and 1.09 per cent w/v of sodium acetate and
587
HYDROXYPROGESTERONE HEXANOATE IP 2007
measure the absorbance of the resulting solution at the Place the dry plate in a tank containing a shallow layer of the
maximum at about 351 nm (2.4.7). Calculate the content of solvent mixture, allow the solvent mixture to ascend to the
C62H89CoN13O15P taking 195 as the specific absorbance at top, remove the plate from the tank and allow the solvent to
351 nm. evaporate. Use within 2 hours, with the flow of the mobile
Storage. Store protected from light. phase in the direction in which the aforementioned treatment
was done.
equivalent amount of anhydrous hydroxocobalamin.
Hydroxyprogesterone Hexanoate Heat at 120° for a further 10 minutes, allow to cool and examine
Hydroxyprogesterone Caproate spot in the chromatogram obtained with the test solution
O CH3 reference solution (a). The principal spot in the chromatogram
H 3C obtained with reference solution (b) appears as a single,
O CH3
compact spot.
H3C H
O C. Dissolve 1 mg in 1 ml of sulphuric acid and allow to stand
H H for 2 minutes; a faint yellow colour is produced which, on the
O addition of 0.5 ml of water, changes first to green, then to red
and finally to reddish-violet with a blue fluorescence.
C27H40O4 Mol. Wt. 428.6
D. Heat 50 mg with 2 ml of 0.5 M ethanolic potassium
Hydroxyprogesterone Hexanoate is 3,20-dioxopregn-4-en- hydroxide in a water-bath for 5 minutes. Add 3 ml of water,
17α-yl hexanoate evaporate the ethanol, add 2 ml of sulphuric acid (50 per
Hydroxyprogesterone Hexanoate contains not less than cent) and heat on a water-bath; the odour of hexanoic acid is
97.0 per cent and not more than 103.0 per cent of C27H40O4, produced.
calculated on the dried basis. Tests
Acidity. Dissolve 0.2 g in 25 ml of ethanol previously
neutralised to bromothymol blue solution and titrate
Identification immediately with 0.01 M sodium hydroxide until a faint blue
colour is produced. Not more than 1.0 ml of 0.01 M sodium
A. Determine by infrared absorption spectrophotometry (2.4.6). hydroxide is required.
Specific optical rotation (2.4.22). +44.0 ° to +49.0°, determined
hydroxyprogesterone hexanoate RS or with the reference
spectrum of hydroxyprogesterone hexanoate.
Related foreign steroids. Determine by thin-layer
HF254.
Solvent mixture. A mixture of 90 volumes of acetone and 10
Mobile phase. A mixture of equal volumes of cyclohexane
volumes of 1,2-propane diol.
and ethyl acetate.
Mobile phase. A mixture of equal volumes of cyclohexane
and light petroleum (40° to 60°).
examination in 10 ml of the same solvent mixture.
with chloroform.
Reference solution (a). Dissolve 25 mg of hydroxy-
progesterone hexanoate RS in 10 ml of the same solvent
mixture.
Reference solution (b). Mix equal volumes of the test solution test solution is not more intense than the spot in the
and reference solution (a). chromatogram obtained with the reference solution.
588
IP 2007 HYDROXYPROPYL CELLULOSE
Sulphated ash (2.3.18). Not more than 0.1 per cent. Tests
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Other tests. Complies with the tests stated under Parenteral
on 2.0 g by drying over phosphorus pentoxide at a pressure Preparations (Injections).
Assay. To a quantity of the injection containing about 0.125 g
Assay. Weigh accurately about 60 mg and dissolve in sufficient of Hydroxyprogesterone Hexanoate add sufficient chloroform
ethanol to produce 100.0 ml. Dilute 5.0 ml to 250.0 ml with to produce 100.0 ml. Dilute 5.0 ml to 100.0 ml with chloroform;
ethanol and measure the absorbance of the resulting solution to 5.0 ml add 10 ml of isoniazid solution and sufficient
at the maximum at about 240 nm (2.4.7). Calculate the content methanol to produce 20.0 ml. Allow to stand for 45 minutes
of C27H40O4 taking 395 as the specific absorbance at 240 nm. and measure the absorbance of the resulting solution at the
Storage. Store protected from light. maximum at about 380 nm (2.4.7), using as the blank 5 ml of
chloroform treated in the same manner. Calculate the content
of C27H40O4 from the absorbance obtained by repeating the
operation using a 0.00625 per cent w/v solution of
Hydroxyprogesterone Injection hydroxyprogesterone hexanoate RS in chloroform and
beginning at the words “to 5.0 ml add...”
Hydroxyprogesterone Caproate Injection;
Hydroxyprogesterone Hexanoate Injection Storage. Store protected from light.
Hydroxyprogesterone Injection is a sterile solution of Labelling. The label states that the preparation is intended
Hydroxyprogesterone Hexanoate in a suitable ester, in a for intramuscular injection only.
suitable fixed oil or in any mixture of fixed oils.
Hydroxyprogesterone Hexanoate Injection contains not less
than 92.5 per cent and not more than 107.5 per cent of the Hydroxypropyl Cellulose
stated amount of hydroxyprogesterone hexanoate, C27H40O4.
Cellulose, 2-Hydroxypropyl Ether; Hyprolose
Identification Hydroxypropylcellulose is a cellulose having some of the
hydroxyl groups in the form of the 2-hydroxypropyl ether. It
may contain not more than 0.6 per cent of silica (SiO2). The
the plate with silica gel HF254.
various grades commercially available are distinguished by a
Mobile phase. A mixture of equal volumes of cyclohexane number indicative of the apparent viscosity in millipascal
and ethyl acetate. seconds of a 2 per cent w/w solution measured at 20°.
Test solution. Dilute the injection with chloroform to give a Description. A white or yellowish white powder; practically
solution containing 1.0 per cent w/v of Hydroxyprogesterone odourless; hygroscopic after drying.
Hexanoate.
Identification
Reference solution. A solution containing 1 per cent w/v of
hydroxyprogesterone hexanoate RS in chloroform. A. With constant stirring add a quantity equivalent to 1 g of
the dried substance into 50 ml of carbon dioxide-free water
Apply to the plate 1µl of each solution. After development,
previously heated to 90°. Allow to cool, dilute to 100 ml with
carbon dioxide-free water and continue stirring until solution
is complete (solution A). Heat 10 ml of solution A on a water-
bath with stirring. At temperatures above 40° the solution
with the reference solution. Ignore any spots due to the
becomes cloudy or a flocculent precipitate is formed. On
vehicle.
cooling, the solution becomes clear.
B. Dissolve a volume of the injection containing 0.1 g of
B. To 10 ml of solution A add 0.3 ml of 2 M acetic acid and 2.5
Hydroxyprogesterone Hexanoate in 10 ml of light petroleum
ml of a 10 per cent w/v solution of tannic acid; a yellowish
(40° to 60°) and extract with three quantities, each of 10 ml, of
white, flocculent precipitate is produced which dissolves in 6
a mixture of 7 volumes of glacial acetic acid and 3 volumes of
M ammonia.
water. Wash the combined extracts with 10 ml of light
petroleum (40 ° to 60°), dilute with water until the solution C. Without heating completely dissolve 0.2 g in 15 ml of a 70
becomes turbid and allow to stand in ice for about 2 hours per cent w/w solution of sulphuric acid, pour the solution
until a white precipitate is produced. The precipitate, after with stirring into 100 ml of iced water. In a test-tube kept in ice,
washing with water, melts at about 120° (2.4.21). mix thoroughly 1 ml of the solution with 8 ml of sulphuric
589
HYDROXYPROPYLMETHYLCELLULOSE IP 2007
acid, added dropwise. Heat in a water-bath for exactly 3 minutes viscosity the label also states the concentration of the solution
and cool immediately in ice. When the mixture is cool, carefully to be used and the apparent viscosity in millipascal seconds.
add 0.6 ml of a solution containing 3 g of ninhydrin in 100 ml
of a 4.55 per cent w/v solution of sodium metabisulphite, mix
well and allow to stand at 25°; a pink colour is produced
immediately which becomes violet within 100 minutes.
Hydroxypropylmethylcellulose
D. Place 1 ml of solution A on a glass plate. After evaporation
of the water a thin film is produced. Cellulose, 2-Hydroxypropylmethyl Ether;
Hypromellose
Tests Hydroxypropylmethylcellulose is a cellulose having some of
Appearance of solution. Solution A is not more opalescent the hydroxyl groups in the form of the methyl ether and some
than opalescence standard OS3 (2.4.1), and not more intensely in the form of the 2-hydroxypropyl ether. The various grades
coloured than reference solution YS6 (2.4.1). commercially available are distinguished by a number
indicative of the apparent viscosity in millipascal seconds of
pH (2.4.24). 5.0 to 8.5, determined in solution A. a 2 per cent w/v solution measured at 20°.
Apparent viscosity. 75 to 140 per cent of the stated value, Description. A white or yellowish white, fibrous or granular
determined by the following method. Weigh accurately a powder; almost odourless; hygroscopic after drying.
quantity equivalent to 2.0 g of the dried substance and add,
with constant stirring, to 50 ml of water previously heated to Identification
90°. Allow to cool, dilute to 100 ml with water and continue
stirring until solution is complete. Adjust the weight of the A. With constant stirring add a quantity containing 1 g of the
solution to 100 g and centrifuge the solution to expel any dried substance into 50 ml of carbon dioxide-free water
trapped air. Determine the viscosity, Method C, at 20° using a previously heated to 90°. Allow to cool, dilute to 100 ml with
shear rate of 10 s-1 (2.4.28). carbon dioxide-free water and continue stirring until solution
is complete (solution A). Heat 10 ml of solution A in a water-
Heavy metals (2.3.13). 1.0 g complies with the limit test for bath with stirring. At temperatures above 50° the solution
heavy metals, Method B (20 ppm). becomes cloudy or a flocculent precipitate is formed. On
Chlorides (2.3.12). Dilute 5.0 ml of solution A to 15 ml with cooling, the solution becomes clear or slightly opalescent.
water. The resulting solution complies with the limit test for B. To 10 ml of solution A add 10 ml of 1 M sodium hydroxide
chlorides (0.5 per cent). or 1 M hydrochloric acid; in either case the mixture remains
Silica. Not more than 0.6 per cent, determined by the following stable.
method. To the residue obtained in the test for Sulphated ash C. To 10 ml of solution A add 0.3 ml of 2 M acetic acid and 2.5
add sufficient ethanol (95 per cent) to moisten the residue ml of a 10 per cent w/v solution of tannic acid; a yellowish
completely. Add 6 ml of hydrofluoric acid in small portions. white, flocculent precipitate is produced which dissolves in
Evaporate to dryness at 95° to 105° taking care to avoid loss 6 M ammonia.
by spurting. Cool and rinse the walls of the platinum crucible
with 6 ml of hydrofluoric acid. Add 0.5 ml of sulphuric acid D. Without heating completely dissolve 0.2 g in 15 ml of a 70
and evaporate to dryness. Progressively increase the per cent w/w solution of sulphuric acid, pour the solution
temperature, ignite at 900°, allow to cool in a desiccator and with stirring into 100 ml of iced water. In a test-tube kept in ice,
weigh. The difference between the weight of the residue mix thoroughly 1 ml of the solution with 8 ml of sulphuric
obtained in the test for Sulphated ash and the weight of the acid, added dropwise. Heat in a water-bath for exactly 3 minutes
final residue is equal to the amount of silica in the substance and cool immediately in ice. When the mixture is cool, carefully
under examination. add 0.6 ml of a solution containing 3 g of ninhydrin in 100 ml
of a 4.55 per cent w/v solution of sodium metabisulphite, mix
Sulphated ash (2.3.18). Not more than 1.6 per cent, determined well and allow to stand at 25°; a pink colour is produced
on 1.0 g in a platinum crucible. immediately which becomes violet within 100 minutes.
Loss on drying (2.4.19). Not more than 7.0 per cent, determined E. Place 1 ml of solution A on a glass plate. After evaporation
on 0.5 g by drying in an oven at 105°. of the water a thin film is produced.
Tests
Labelling. The label states the apparent viscosity in millipascal
seconds of a 2 per cent w/w solution. For products of low pH (2.4.24). 5.5 to 8.0, determined in solution A.
590
IP 2007 HYOSCINE BUTYLBROMIDE
Appearance of solution. Solution A is not more opalescent Identification

than opalescence standard OS3 (2.4.1), and not more intensely
coloured than reference solution YS6 (2.4.1). Test A may be omitted if tests B, C and D are carried out. Tests
C and D may be omitted if tests A and B are carried out.
Apparent viscosity. 75 to 140 per cent of the stated value,
determined by the following method. Weigh accurately a A. Determine by infrared absorption spectrphotometry (2.4.6).
quantity equivalent to 2.0 g of the dried substance and add, Compare the spectrum with that obtained with hyoscine
with constant stirring, to 50 ml of water previously heated to butylbromide RS or with the reference spectrum of hyoscine
90°. Allow to cool, dilute to 100 ml with water and continue butylbromide.
stirring until solution is complete. Adjust the weight of the B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.15
solution to 100 g and centrifuge the solution to expel any per cent w/v solution in 0.01 M hydrochloric acid shows
trapped air. Determine the viscosity, Method C, at 20° using a absorption maxima at about 252 nm, 257 nm and 264 nm and a
shear rate of 10 s-1 (2.4.28). less well-defined maximum at about 247 nm; absorbance at
Heavy metals (2.3.12). 1.0 g complies with the limit test for about 252 nm, about 0.50, at about 257 nm, about 0.67 and at
heavy metals, Method B (20 ppm). about 264 nm, about 0.50.
Chlorides (2.3.12). Dilute 5.0 ml of solution A to 15 ml with C. To 1 mg add 0.2 ml of fuming nitric acid and evaporate to
water. The resulting solution complies with the limit test for dryness on a water-bath. Dissolve the residue in 2 ml of acetone
chlorides (0.5 per cent). and add 0.1 ml of a 3 per cent w/v solution of potassium
hydroxide in methanol; a violet colour is produced.
Loss on drying (2.4.19). Not more than 10.0 per cent, D. Gives the reactions of bromides (2.3.1).
determined on 0.5 g by drying in an oven at 105°.
Tests
Labelling. The label states the apparent viscosity in millipascal
seconds of a 2 per cent w/v solution.
Specific optical rotation (2.4.22). –l8.0° to –20.0°, determined
Hyoscine Butylbromide
Apo-compounds. Ratio of the absorbance (2.4.7) of a 0.1 per
Scopolamine Butylbromide cent w/v solution in 0.01 M hydrochloric acid at the maximum
at about 247 nm to that at the maximum at about 264 nm is not
more than 0.94.
CH3
H3C N Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with microcrystalline cellulose.
O
Mobile phase. The upper layer obtained by shaking together
H OH
50 volumes of 1-butanol, 25 volumes of water and 5 volumes
Br of anhydrous formic acid.
O
O
C21H30BrNO4 Mol. Wt. 440.4 w/v of the substance under examination in methanol (50 per
Hyoscine Butylbromide is (1S,3s,5R,6R,7S,8r)-6,7-epoxy-8- cent).
butyl-3-[(S)-tropoyloxy]tropanium bromide. Reference solution (b). A solution containing 0.002 per cent
Hyoscine Butylbromide contains not less than 98.0 per cent w/v of hyoscine hydrobromide RS in methanol (50 per cent).
and not more than 101.0 per cent of C21H30BrNO4, calculated Apply to the plate 10 µl of each solution. After development,
on the dried basis. dry the plate in air and spray with dilute potassium
Description. A white or almost white, crystalline powder; iodobismuthate solution. In the chromatogram obtained with
odourless or almost odourless. the test solution any spot corresponding to hyoscine
591
HYOSCINE BUTYLBROMIDE INJECTION IP 2007
hydrobromide is not more intense than the spot in the and add 0.1 ml of a 3 per cent w/v solution of potassium
chromatogram obtained with reference solution (b) and any hydroxide in methanol; a violet colour is produced.
other secondary spot is not more intense than the spot in the
chromatogram obtained with reference solution (a). Tests
Sulphated ash (2.3.18). Not more than 0.1 per cent. pH (2.4.24). 3.7 to 5.5.
Loss on drying (2.4.19). Not more than 2.5 per cent, determined Related substances. Determine by thin-layer chromatography
on 0.5 g by drying in an oven at 105°. (2.4.17), coating the plate with microcrystalline cellulose.
Assay. Weigh accurately about 0.6 g, dissolve in 30 ml of Mobile phase. The upper layer obtained by shaking together
anhydrous glacial acetic acid and add 5 ml of mercuric 50 volumes of 1-butanol, 25 volumes of water and 5 volumes
acetate solution. Titrate with 0.1 M perchloric acid, using 1- of anhydrous formic acid.
naphtholbenzein solution as indicator. Carry out a blank
Test solution. The injection diluted if necessary with methanol
titration.
(50 per cent) to contain 2 per cent w/v of Hyoscine
1 ml of 0.1 M perchloric acid is equivalent to 0.04404 g of Butylbromide.
C21H30BrNO4.
Storage. Store at a temperature not exceeding 30° protected 20 volumes with methanol (50 per cent).
from light and moisture.
w/v of hyoscine hydrobromide RS in methanol (50 per cent).
Hyoscine Butylbromide Injection iodobismuthate solution. In the chromatogram obtained with
Scopolamine Butylbromide Injection the test solution any spot corresponding to hyoscine
hydrobromide is not more intense than the spot in the
Hyoscine Butylbromide Injection is a sterile solution of chromatogram obtained with reference solution (b) and any
Hyoscine Butylbromide in Water for Injections. other secondary spot is not more intense than the spot in the
Hyoscine Butylbromide Injection contains not less than 92.5 chromatogram obtained with reference solution (a).
per cent and not more than 107.5 per cent of the stated amount Other tests. Complies with the tests stated under Parenteral
of hyoscine butylbromide, C21H30BrNO4. Preparations (Injections).
Assay. Dilute a volume of the injection containing about 60
Identification
mg of Hyoscine Butylbromide to 50.0 ml with water and dilute
Evaporate to dryness a volume of the injection containing 0.1 5.0 ml of this solution to 200.0 ml with water. To 10.0 ml of the
g of Hyoscine Butylbromide, shake the residue with 20 ml of resulting solution add 10 ml of water, 15 ml of dichloromethane,
chloroform, filter, evaporate the filtrate to dryness and triturate 15 ml of a 0.01 per cent w/v solution of hexanitrodiphenylamine
the residue with 5 ml of acetonitrile. Evaporate to dryness in dichloromethane and 5 ml of 5 M sodium hydroxide and
and dry the residue at 50° at a pressure not exceeding 0.7 kPa shake for 2 minutes. Allow the layers to separate and reserve
for 1 hour. The residue complies with the following tests. the organic layer. Extract the aqueous layer with successive
quantities, each of 5 ml, of dichloromethane until no further
A. Determine by infrared absorption spectrphotometry (2.4.6).
colour is extracted from the aqueous layer. Add the
Compare the spectrum with that obtained with hyoscine
dichloromethane extracts to the reserved organic layer, filter
butylbromide RS or with the reference spectrum of hyoscine
through absorbent cotton, add sufficient dichloromethane
butylbromide.
to produce 50.0 ml and measure the absorbance of the resulting
B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.15 solution at the maximum at about 420 nm (2.4.7), using as the
per cent w/v solution in 0.01 M hydrochloric acid shows blank a solution prepared by repeating the procedure without
absorption maxima at about 252 nm, 257 nm and 264 nm and a the injection. Calculate the content of C21H30BrNO4 from the
less well-defined maximum at about 247 nm; absorbance at absorbance obtained by repeating the procedure using 10.0
about 252 nm, about 0.50, at about 257 nm, about 0.67 and at ml of a 0.003 per cent w/v solution of hyoscine butylbromide
about 264 nm, about 0.50. RS and beginning at the words “add 10 ml of water....”.
C. To 1 mg add 0.2 ml of fuming nitric acid and evaporate to Storage. Store at a temperature not exceeding 30° protected
dryness on a water-bath. Dissolve the residue in 2 ml of acetone from light and moisture.
592
IP 2007 HYOSCINE BUTYLBROMIDE TABLETS
Hyoscine Butylbromide Tablets hydrobromide is not more intense than the spot in the
chromatogram obtained with reference solution (b) and any
Scopolamine Butylbromide Tablets other secondary spot is not more intense than the spot in the
Hyoscine Butylbromide Tablets contain not less than 92.5 per chromatogram obtained with reference solution (a).
cent and not more than 107.5 per cent of the stated amount of Uniformity of content. Comply with the test stated under
hyoscine butylbromide, C21H30BrNO4. The tablets may be Tablets.
coated.
Shake one tablet with 100 ml of water for 30 minutes, add
Identification sufficient water to produce 250.0 ml and filter. To 10.0 ml of the
resulting solution add 10 ml of water, 15 ml of dichloromethane,
Shake a quantity of the powdered tablets containing 50 mg of 15 ml of a 0.01 per cent w/v solution of hexanitrodiphenylamine
Hyoscine Butylbromide with 20 ml of chloroform, filter, in dichloromethane and 5 ml of 5 M sodium hydroxide and
evaporate the filtrate to dryness and triturate the residue with shake for 2 minutes. Allow the layers to separate and reserve
5 ml of acetonitrile. Evaporate to dryness and dry the residue the organic layer. Extract the aqueous layer with successive
at 50° at a pressure not exceeding 0.7 kPa for 1 hour. The quantities, each of 5 ml, of dichloromethane until no further
residue complies with the following tests. colour is extracted from the aqueous layer. Add the
A. Determine by infrared absorption spectrphotometry (2.4.6). dichloromethane extracts to the reserved organic layer, filter
Compare the spectrum with that obtained with hyoscine through absorbent cotton, add sufficient dichloromethane
butylbromide RS or with the reference spectrum of hyoscine to produce 50.0 ml and measure the absorbance of the resulting
butylbromide. solution at the maximum at about 420 nm (2.4.7), using as the
blank a solution prepared by repeating the procedure without
B. When examined in the range 230 nm to 360 nm, a 0.15 per the injection. Calculate the content of C21H30BrNO4 from the
cent w/v solution in 0.01 M hydrochloric acid shows absorbance obtained by repeating the procedure using
absorption maxima at about 252 nm, 257 nm and 264 nm and a 10.0 ml of a 0.003 per cent w/v solution of hyoscine
less well-defined maximum at about 247 nm; absorbance at butylbromide RS and beginning at the words “add 10 ml of
about 252 nm, about 0.50, at about 257 nm, about 0.67 and at
water....”.
about 264 nm, about 0.50 (2.4.7).
C. To 1 mg add 0.2 ml of fuming nitric acid and evaporate to
dryness on a water-bath. Dissolve the residue in 2 ml of acetone Assay. Weigh and powder 20 tablets. Weigh accurately a
and add 0.1 ml of a 3 per cent w/v solution of potassium quantity of the powder containing about 3 mg of Hyoscine
hydroxide in methanol; a violet colour is produced. Butylbromide and shake with 50 ml of water for 30 minutes,
add sufficient water to produce 100.0 ml and filter. To 10.0 ml
Tests of the resulting solution add 10 ml of water, 15 ml of
dichloromethane, 15 ml of a 0.01 per cent w/v solution of
hexanitrodiphenylamine in dichloromethane and 5 ml of 5 M
sodium hydroxide and shake for 2 minutes. Allow the layers
Mobile phase. The upper layer obtained by shaking together to separate and reserve the organic layer. Extract the aqueous
50 volumes of 1-butanol, 25 volumes of water and 5 volumes layer with successive quantities, each of 5 ml, of
of anhydrous formic acid. dichloromethane until no further colour is extracted from the
Test solution. Shake a quantity of the powdered tablets aqueous layer. Add the dichloromethane extracts to the
containing 40 mg of Hyoscine Butylbromide with 2 ml of reserved organic layer, filter through absorbent cotton, add
methanol (50 per cent) and filter. sufficient dichloromethane to produce 50.0 ml and measure
Reference solution (a). Dilute 1 volume of the test solution to about 420 nm (2.4.7), using as the blank a solution prepared
500 volumes with methanol (50 per cent). by repeating the procedure without the injection. Calculate
Reference solution (b). A solution containing 0.002 per cent the content of C21H30BrNO4 from the absorbance obtained by
w/v of hyoscine hydrobromide RS in methanol (50 per cent). repeating the procedure using 10.0 ml of a 0.003 per cent w/v
solution of hyoscine butylbromide RS and beginning at the
Apply to the plate 10 µl of each solution. After development, words “add 10 ml of water....”.
iodobismuthate solution. In the chromatogram obtained with Storage. Store at a temperature not exceeding 30° protected
the test solution any spot corresponding to hyoscine from light and moisture.
593
HYOSCINE HYDROBROMIDE IP 2007
Hyoscine Hydrobromide Mobile phase. A mixture of 50 volumes of chloroform, 30

volumes of acetone, 10 volumes of methanol and 2 volumes
Scopolamine Hydrobromide of strong ammonia solution.
CH3 examination in 10 ml of methanol.
N
Reference solution (a). Dilute 1 ml of the test solution to 100
O ml with methanol.
H OH Reference solution (b). Dilute 25 ml of reference solution (a)
to 50 ml with methanol.
O , HBr , 3H2O
O dry the plate at 105° for 15 minutes, allow to cool and spray
with dilute potassium iodobismuthate solution until spots
appear. Any secondary spot in the chromatogram obtained
C17H21NO4,HBr,3H2O Mol. Wt. 438.3 with the test solution is not more intense than the spot in the
Hyoscine Hydrobromide is (1S,3s,5R,6R,7S)-6,7-
epoxytropan-3-yl-(S)-tropate hydrobromide trihydrate.
chromatogram obtained with reference solution (b). Ignore
Hyoscine Hydrobromide contains not less than 99.0 per cent any yellow spot remaining on the line of application.
and not more than 101.0 per cent of C17H21NO4,HBr, calculated
Description. Colourless crystals or a white, crystalline powder; Water (2.3.43). 10.0 to 13.0 per cent, determined on 0.2 g.
odourless; efflorescent. Assay. Weigh accurately about 0.4 g, dissolve in 10 ml of
anhydrous glacial acetic acid, warming if necessary, cool
Identification the solution and add 20 ml of dioxan. Titrate with 0.1 M
Test A may be omitted if tests B, C and D are carried out. Tests perchloric acid, determining the end-point potentiometrically
B and C may be omitted if tests A and D are carried out. (2.4.25). Carry out a blank titration.
A.Determine by infrared absorption spectrophotometry (2.4.6). 1 ml of 0.1 M perchloric acid is equivalent to 0.03843 g of
Compare the spectrum with that obtained with hyoscine C17H21NO4,HBr.
hydrobromide RS. Storage. Store protected from light and moisture in well-filled
B. To about 1 mg add 0.2 ml of fuming nitric acid and evaporate containers of small capacity in a refrigerator at a temperature
to dryness on a water-bath. Dissolve the residue in 2 ml of not exceeding 15°.
acetone and add 0.1 ml of a 3 per cent w/v solution of potassium
hydroxide in methanol; a violet colour is produced.
C. Gives the reaction of alkaloids (2.3.1).
Hyoscine Hydrobromide Injection
D. Gives the reactions of bromides (2.3.1).
Scopolamine Hydrobromide Injection
Tests
Hyoscine Hydrobromide Injection is a sterile solution of
pH (2.4.24). 4.0 to 5.5, determined in a 5.0 per cent w/v solution. Hyoscine Hydrobromide in Water for Injection.
Specific optical rotation (2.4.22). –24.0° to –27.0°, determined Hyoscine Hydrobromide Injection contains not less than 90.0
in a 5.0 per cent w/v solution. per cent and not more than 110.0 per cent of the stated amount
of hyoscine hydrobromide, C17H21NO4,HBr,3H2O.
Apohyoscine. Dissolve 0.1 g in sufficient 0.01 M hydrochloric
acid to produce 100 ml and measure the absorbance (2.4.7) at
Identification
about 245 nm; absorbance, about 0.36, calculated on the
anhydrous basis (0.5 per cent). A. Determine by thin-layer chromatography (2.4.17), coating
Related substances and decomposition products. Determine the plate with silica gel G.
by thin-layer chromatography (2.4.17), coating the plate with Mobile phase. A mixture of 50 volumes of chloroform, 40
silica gel G. volumes of acetone and 10 volumes of diethylamine.
594
IP 2007 HYOSCINE HYDROBROMIDE TABLETS
Test solution. Evaporate a volume of the injection containing Chromatographic system

5 mg of Hyoscine Hydrobromide to dryness on a water-bath, – a glass column 1.5 m x 4 mm, packed with acid-washed,
triturate the residue with 1 ml of ethanol (95 per cent), allow diatomaceous support (80 to 100 mesh) coated with 3
to stand and use the supernatant liquid. per cent w/w phenyl methyl silicone fluid (50 per cent
Reference solution. A solution containing 0.5 per cent w/v of phenyl),
hyoscine hydrobromide RS in ethanol (95 per cent). – temperature:
column.230°,
Apply to the plate 5 µl of each solution. After development, inlet port and detector. 280°,
dry the plate in air, heat it at 105° for 20 minutes, allow to cool – flow rate. 30 ml per minute of the carrier gas.
and spray with dilute potassium iodobismuthate solution.
The principal spot in the chromatogram obtained with the test Calculate the content of C17H21NO4,HBr,3H2O in the injection.
solution corresponds to that in the chromatogram obtained Storage. Store at a temperature not exceeding 15°, protected
with the reference solution. from light.
B. In the Assay, the chromatogram obtained with test solution
(b) shows a peak with the same retention time as the peak
derived from hyoscine hydrobromide in the chromatogram
obtained with the reference solution. Hyoscine Hydrobromide Tablets
C. Evaporate a suitable volume to dryness. To 1 ml of a 1 per Scopolamine Hydrobromide Tablets
cent w/v solution of the residue add 1 ml of 5 M ammonia,
Hyoscine Hydrobromide Tablets contain not less than 90.0
shake with chloroform and evaporate the chloroform solution
per cent and not more than 110.0 per cent of the stated amount
to dryness on a water-bath. To the residue add 1.5 ml of a 2 per
of hyoscine hydrobromide, C17H21NO4,HBr,3H2O.
cent w/v solution of mercuric chloride in ethanol (60 per
cent); a white precipitate is produced which dissolves on Identification
warming (distinction from atropine and hyoscyamine).
D. Gives reaction A of bromides (2.3.1).
Tests Mobile phase. A mixture of 50 volumes of chloroform, 40
pH (2.4.24). 4.0 to 6.0. volumes of acetone and 10 volumes of diethylamine.
Other tests. Complies with the tests stated under Parenteral Test solution. Shake a quantity of the powdered tablets
Preparations (Injections). containing 10 mg of Hyoscine Hydrobromide with 2 ml of
ethanol (95 per cent) and centrifuge.
Test solution (a). Add 1 of a 0.3 per cent w/v solution of
hyoscine hydrobromide RS in ethanol (95 per cent).
atropine sulphate RS (internal standard) in methanol (solution
A) and 1 ml of 5 M ammonia to a volume of the injection Apply to the plate 5 µl of each solution. After development,
containing about 5 mg of Hyoscine Hydrobromide, dilute if dry the plate in air, heat it at 105° for 20 minutes, allow to cool
necessary to 15 ml with water. Extract with two quantities, and spray with dilute potassium iodobismuthate solution.
each of 10 ml, of chloroform, shake the combined extracts The principal spot in the chromatogram obtained with the test
with 2 g of anhydrous sodium sulphate, filter and evaporate solution corresponds to that in the chromatogram obtained
the filtrate to dryness. Dissolve the residue in 2.0 ml of with the reference solution.
dichloromethane. To 1.0 ml of this solution add 0.2 ml of a B. In the Assay, the chromatogram obtained with test solution
mixture of 4 volumes of N,O-bis(trimethylsilyl)acetamide and (b) shows a peak with the same retention time as the peak
1 volume of trimethylchlorosilane, mix and allow to stand for derived from hyoscine hydrobromide in the chromatogram
30 minutes. obtained with the reference solution.
Test solution (b). Prepare in the same manner as test solution C. Extract a quantity of the powdered tablets containing 1 mg
(a) but omitting the addition of solution A. of Hyoscine Hydrobromide with 5 ml of ethanol (95 per cent),
Reference solution. Add 1 ml of solution A and 1 ml of 5 M filter and evaporate the filtrate to dryness on a water-bath.
ammonia to 15.0 ml of a 0.033 per cent w/v solution of hyoscine Cool, add 0.2 ml of fuming nitric acid and again evaporate to
hydrobromide RS and complete the procedure described dryness on a water-bath; a yellow residue is produced. To the
under test solution (a) beginning at the words “Extract with cooled residue add 2 ml of acetone and 0.2 ml of a 3 per cent w/
two quantities,....”. v solution of potassium hydroxide in methanol; a deep violet
595
HYOSCINE HYDROBROMIDE TABLETS IP 2007
colour is produced. (Atropine and hyoscyamine also yield sulphate RS.

this reaction; the reaction is masked by other alkaloids). Other tests. Comply with the tests stated under Tablets.
D. The powdered tablets give reaction A of bromides (2.3.1).
Tests Test solution (a). Shake a quantity of the powdered tablets
containing about 5 mg of Hyoscine Hydrobromide with 10 ml
Uniformity of content. Comply with the test stated under of 0.1 M hydrochloric acid. Add 1 ml of a 0.3 per cent w/v
Tablets. solution of atropine sulphate RS (internal standard) in
For tablets containing 600 µg of Hyoscine Hydrobromide - methanol (solution A), extract with two quantities, each of
10 ml, of chloroform and discard the chloroform extracts. Add
Test solution (a). Powder one tablet and triturate with 5 ml of
1 ml of 5 M ammonia. Extract with two quantities, each of
0.1 M hydrochloric acid. Add 1.0 ml of a 0.0375 per cent w/v
10 ml, of chloroform, shake the combined extracts with 2 g of
solution of atropine sulphate RS (internal standard) in
anhydrous sodium sulphate, filter and evaporate the filtrate
methanol (solution A), extract with two quantities, each of 5
to dryness. Dissolve the residue in 2.0 ml of dichloromethane.
ml, of chloroform and discard the chloroform extracts. Add 1
To 1.0 ml of this solution add 0.2 ml of a mixture of 4 volumes
ml of 5 M ammonia. Extract with two quantities, each of 5 ml,
of N,O-bis(trimethylsilyl)acetamide and 1 volume of
of chloroform, shake the combined extracts with 1 g of
trimethylchlorosilane, mix and allow to stand for 30 minutes.
anhydrous sodium sulphate, filter and evaporate the filtrate
to dryness. Dissolve the residue in 0.5 ml of a mixture of 20 Test solution (b). Prepare in the same manner as test solution
volumes of dichloromethane, 4 volumes of N,O-bis(tri- (a) but omitting the addition of solution A.
methylsilyl) acetamide and 1 volume of trimethyl- Reference solution. Add 1 ml of solution A and 1 ml of 5 M
chlorosilane, mix and allow to stand for 30 minutes. ammonia to 15.0 ml of a 0.033 per cent w/v solution of hyoscine
Test solution (b). Prepare in the same manner as test solution hydrobromide RS and complete the procedure described
(a) but omitting the addition of solution A. under test solution (a) beginning at the words “Extract with
two quantities,”
Reference solution. Add 1 ml of a 0.0375 per cent w/v solution
of atropine sulphate RS (internal standard) in methanol Chromatographic system
(solution A) and 1 ml of 5 M ammonia to 5.0 ml of a 0.012 per – a glass column 1.5 m x 4 mm, packed with acid-washed,
cent w/v solution of hyoscine hydrobromide RS and complete diatomaceous support (80 to 100 mesh) coated with 3
the procedure described under test solution (a) beginning at per cent w/w phenyl methyl silicone fluid (50 per cent
the words “extract with two quantities,...”. phenyl),
– temperature:
Carry out the procedure described in the Assay. Calculate the
column.230°,
content of C17H21NO4,HBr,3H2O in the tablet.
For tablets containing less than 600 µg of Hyoscine – flow rate. 30 ml per minute of the carrier gas.
Hydrobromide -
Calculate the content of C17H21NO4,HBr,3H2O in the tablets.
Use the same procedure but with correspondingly smaller
Storage. Store at a temperature not exceeding 15°, protected
concentrations of hyoscine hydrobromide RS and atropine
from light.
596
I
Ibuprofen ....
Ibuprofen Tablets ....
Idoxuridine ....
Idoxuridine Eye Drops ....
Imipenem ....
Imipenem and Cilastatin Injection ....
Imipramine Hydrochloride ....
Imipramine Tablets ....
Indinavir Sulphate ....
Indinavir Capsules ....
Indomethacin ....
Indomethacin Capsules ....
Indomethacin Suppositories ....
Insulin ....
Human Insulin ....
Insulin Injection ....
Biphasic Insulin Injection ....
Biphasic Isophane Insulin Injection ....
Isophane Insulin Injection ....
Protamine Zinc Insulin Injection ....
Insulin Zinc Suspension ....
Insulin Zinc Suspension (Amorphous) ....
Insulin Zinc Suspension (Crystalline) ....
Invert Sugar Injection ....
Invert Sugar And Sodium Chloride Injection ....
Iodine ....
Ipratropium Bromide ....
Irinotecan Hydrochloride Trihydrate ....
Irinotecan Injection ....
Iron and Ammonium Citrate ....
597
Iron Dextran Injection ....

Isoniazid ....
Isoniazid Tablets ....
Isoprenaline Hydrochloride ....
Isoprenaline Injection ....
Isoprenaline Sulphate ....
Isoprenaline Tablets ....
Isopropyl Alcohol ....
Diluted Isosorbide Dinitrate ....
Isosorbide Dinitrate Tablets ....
Isoxsuprine Hydrochloride ....
Isoxsuprine Injection ....
Isoxsuprine Tablets ....
598
IP 2007 IBUPROFEN
Ibuprofen Optical rotation (2.4.22). +0.05° to –0.05°, determined in a

2.5 per cent w/v solution in methanol.
CH3 Related substances. Determine by liquid chromatography
(2.4.14).
CH3 COOH
H3 C examination in 2 ml of acetonitrile and add sufficient of the
mobile phase to produce 10 ml.
C13H18O2 Mol. Wt. 206.3 Reference solution (a). Dilute 1 ml of the test solution to 100
Ibuprofen is (RS)-2-(4-isobutylphenyl)propionic acid.
Reference solution (b). Dissolve 20 mg of ibuprofen RS in 2
Ibuprofen contains not less than 98.5 per cent and not more ml of acetonitrile, add 1 ml of 0.006 per cent w/v solution of 2-
than 101.0 per cent of C13H18O2, calculated on the dried basis. (4-butylphenyl)propionic acid RS in acetonitrile and add
Description. A white or almost white, crystalline powder or sufficient mobile phase to produce 10 ml.
colourless crystals; odour, slight. Chromatographic system
Identification octadecylsilyl silica gel (5 µm),
Test A may be omitted if tests B and C are carried out. Tests B – mobile phase: a mixture of 600 volumes of water, 340
and C may be omitted if test A is carried out. volumes of acetonitrile and 0.5 volume of phosphoric
acid diluted to 1000 volumes with water after
A. Determine by infrared absorption spectrophotometry (2.4.6). equilibration,
Compare the spectrum with that obtained with ibuprofen RS – flow rate. 2 ml per minute,
or with the reference spectrum of ibuprofen. – spectrophotometer set at 214 nm,
B. When examined in the range 230 nm to 360 nm (2.4.7), a – a 20 µl loop injector.
0.05 per cent w/v solution in 0.1 M sodium hydroxide shows Adjust the sensitivity so that the height of the principal peak
absorption maxima at about 264 nm and 272 nm, and a shoulder in the chromatogram obtained with reference solution (a) is
at about 258 nm. The ratio of the absorbance at about 264 nm about 80 per cent of full-scale deflection on the recorder. Record
to that at the shoulder at about 258 nm is 1.20 to 1.30. The ratio the chromatogram for 1.5 times the retention time of the
of the absorbance at the maximum at about 272 nm to that at principal peak. Equilibrate the column with the mobile phase
the shoulder at about 258 nm is 1.00 to 1.10. for about 45 minutes before starting the chromatography.
C. Determine by thin-layer chromatography (2.4.17), coating The retention time of ibuprofen is about 20 minutes. In the
the plate with silica gel H. chromatogram obtained with reference solution (b) measure
Mobile phase. A mixture of 75 volumes of n-hexane, 25 volumes the height (a) of the peak due to 2-(4-butylphenyl) propionic
of ethyl acetate and 5 volumes of glacial acetic acid. acid and the height (b) of the lowest point of the curve
separating this peak from that due to ibuprofen. The test is
Test solution. Dissolve 0.5 g of the substance under not valid unless a is greater than 1.5b. If necessary, adjust the
examination in 100 ml of dichloromethane. concentration of acetonitrile in the mobile phase to obtain the
Reference solution. A 0.5 per cent w/v solution of ibuprofen required resolution. Verify the repeatability by making five
RS in dichloromethane. separate injections of 20 µl of reference solution (a). The test
is not valid unless the relative standard deviation of the area
of the principal peak is less than 2.0 per cent.
dry the plate at 120° for 30 minutes, lightly spray the plate with
a 1 per cent w/v solution of potassium permanganate in 1 M In the chromatogram obtained with the test solution, the area
sulphuric acid, heat at 120° for 20 minutes and examine in of any peak corresponding to 2-(4-butylphenyl) propionic acid
ultraviolet light at 365 nm. The principal spot in the is not greater than that of the peak due to 2-(4-butylphenyl)
chromatogram obtained with the test solution corresponds to propionic acid in the chromatogram obtained with reference
that in the chromatogram obtained with the reference solution. solution (b), the area of any other secondary peak is not greater
than 0.3 times the area of the principal peak in the chromatogram
Tests obtained with reference solution (a) and the sum of the areas
of any such peaks is not greater than 0.7 times the area of the
Appearance of solution. A 10.0 per cent w/v solution in ethanol principal peak in the chromatogram obtained with reference
(95 per cent) is clear (2.4.1), and colourless (2.4.1). solution (a). Ignore any peak the area of which is less than
599
IBUPROFEN TABLETS IP 2007
0.1 times the area of the principal peak in the chromatogram light at 365 nm. Any secondary spot in the chromatogram
obtained with reference solution (a). obtained with the test solution is not more intense than the
Heavy metals (2.3.13). 2.0 g complies with the limit test for spot in the chromatogram obtained with the reference solution.
heavy metals, Method B (10 ppm). Ignore any spot with an Rf value relative to ibuprofen of about
1.2.
Apparatus. No 1
on 1.0 g by drying over phosphorus pentoxide at a pressure
not exceeding 0.1 kPa. Medium. 900 ml of phosphate buffer pH 7.2
ethanol (95 per cent) and titrate with 0.1 M sodium hydroxide Withdraw a suitable volume of the medium and filter promptly
using 0.2 ml of phenolphthalein solution as indicator. Carry through a membrane filter disc having an average pore diameter
out a blank titration. not greater than 1.0 µm, rejecting the first 1 ml of the filtrate.
Dilute a suitable volume of the filtrate with the same solvent.
1 ml of 0.1 M sodium hydroxide is equivalent to 0.02063 g of Measure the absorbance of the resulting solution at the
C13H18O2. maximum at about 221 nm (2.4.7). Calculate the content of
C13H18O2.
D. Not less than 50 per cent of the stated amount of C13H18O2.
Ibuprofen Tablets
Ibuprofen Tablets contain not less than 95.0 per cent and not
more than 105.0 per cent of the stated amount of ibuprofen, Assay. Weigh and powder 20 tablets. Weigh accurately a
C13H18O2. The tablets are coated. quantity of the powder containing about 0.5 g of Ibuprofen,
extract with 60 ml of chloroform for 15 minutes and filter
Identification through a sintered-glass crucible of porosity 3. Wash the
residue with three quantities, each of 10 ml, of chloroform and
A. Extract a quantity of the powdered tablets containing 0.5 g gently evaporate the filtrate just to dryness in a current of air.
of Ibuprofen with 20 ml of acetone, filter and evaporate the Dissolve the residue in 100 ml of ethanol (95 per cent),
filtrate to dryness in a current of air without heating. The previously neutralized to phenolphthalein solution, and titrate
residue complies with the following test. with 0.1 M sodium hydroxide using phenolphthalein solution
Determine by infrared absorption spectrophotometry (2.4.6). as indicator.
Compare the spectrum with that obtained with ibuprofen RS 1 ml of 0.1 M sodium hydroxide is equivalent to 0.02063 g of
or with the reference spectrum of ibuprofen. C13H18O2.
B. The residue obtained in test A, after recrystallisation from
light petroleum (40° to 60°), melts at about 75° (2.4.21).
Tests Idoxuridine
Related substances. Determine by thin-layer chromatography O
I
Mobile phase. A mixture of 75 volumes of n-hexane, 25 volumes HN
of ethyl acetate and 5 volumes of glacial acetic acid.
HO O N
Test solution. Extract a quantity of the powdered tablets O
containing 0.2 g of Ibuprofen with three quantities, each of 10
ml, of chloroform, filter, evaporate the combined filtrate to
about 1 ml and add sufficient chloroform to produce 2 ml. OH
C9H11IN2O5 Mol. Wt. 354.1
volumes with chloroform.
Apply to the plate 5 µl of each solution. After development, Idoxuridine is 2'-deoxy-5-iodouridine.
dry the plate in air, lightly spray the plate with a 1 per cent Idoxuridine contains not less than 98.0 per cent and not more
w/v solution of potassium permanganate in 1 M sulphuric than 101.0 per cent of C9H11IN2O5, calculated on the dried
acid, heat at 120° for 20 minutes and examine in ultraviolet basis.
600
IP 2007 IDOXURIDINE EYE DROPS
Description. A white or almost white, crystalline powder; development, dry it in a current of cold air and examine in
odourless. ultraviolet light at 254 nm. The spots due to 5-iodouracil and
2’-deoxyuridine in the chromatogram obtained with reference
Identification solution (b) are more intense than any corresponding spots in
Test A may be omitted if tests B, C and D are carried out. Tests the chromatogram obtained with test solution (a). Any other
B, C and D may be omitted if test A is carried out. secondary spot in the chromatogram obtained with test
A. Determine by infrared absorption spectrophotometry (2.4.6). chromatogram obtained with reference solution (a).
Compare the spectrum with that obtained with idoxuridine
RS or with the reference spectrum of idoxuridine. Examine the Iodide. Not more than 0.1 per cent, determined by the following
substances as dispersions containing 1 mg in 0.3 g of potassium method. Dissolve 0.25 g in 25 ml of 0.1 M sodium hydroxide,
bromide IR. 5 ml of dilute hydrochloric acid and sufficient water to produce
50 ml, allow to stand for 10 minutes and filter. To 25 ml of the
B. When examined in the range 230 nm to 360 nm (2.4.7), a filtrate add 5 ml of hydrogen peroxide solution (10 vol) and
0.004 per cent w/v solution in 0.01 M sodium hydroxide shows 10 ml of chloroform and shake. Any pink colour produced in
an absorption maximum only at about 279 nm; absorbance at the organic layer is not more intense than that obtained by
about 279 nm, about 0.65. repeating the procedure using 1 ml of a 0.033 per cent w/v
C. In the test for Related substances, the principal spot in the solution of potassium iodide in place of the substance under
chromatogram obtained with test solution (b) corresponds to examination.
that in the chromatogram obtained with reference solution (c). Sulphated ash (2.3.18). Not more than 0.1 per cent.
D. Heat about 5 mg in a test-tube over a naked flame; a violet Loss on drying (2.4.19). Not more than 1.0 per cent, determined
vapour is evolved. on 1.0 g by drying in an oven at 60° over phosphorus pentoxide
Tests at a pressure of 1.5 to 2.5 kPa.
dimethylformamide and titrate with 0.1 M tetrabutyl-
sodium hydroxide is clear (2.4.1), and colourless (2.4.1).
ammonium hydroxide, determining the end-point
pH (2.4.24). 5.5 to 6.5, determined in a 0.1 per cent w/v solution. potentiometrically (2.4.25). Carry out a blank titration.
Specific optical rotation (2.4.22). +28.0° to +32.0°, determined 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
in a 1.0 per cent w/v solution in 1 M sodium hydroxide. 0.03541 g of C9H11IN2O5.
Related substances. Determine by thin-layer chromatography Storage. Store protected from light.
Mobile phase. A mixture of 50 volumes of 2-propanol, 40
volumes of chloroform and 10 volumes of strong ammonia
solution.
Idoxuridine Eye Drops
Test solution (a). Dissolve 0.4 g of the substance under Idoxuridine Eye Drops are a sterile solution of Idoxuridine in
examination in 10 ml of a mixture of 5 volumes of methanol and Purified Water.
1 volume of strong ammonia solution. Idoxuridine Eye Drops contain not less than 90.0 per cent and
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with not more than 110.0 per cent of the stated amount of
the same solvent mixture. idoxuridine, C9H11IN2O5.
Reference solution (a). Dilute 5 ml of test solution (b) to 100 Identification

ml with the same solvent mixture.
A. Dilute a suitable volume with 0.01 M sodium hydroxide to
Reference solution (b). A solution containing 0.02 per cent w/ produce a solution containing 0.003 per cent w/v of
v each of 5-iodouracil RS and 2’-deoxyuridine RS in the Idoxuridine. When examined in the range 230 nm to 360 nm
same solvent mixture. (2.4.7), the resulting solution shows an absorption maximum
Reference solution (c). A solution containing 0.4 per cent w/ only at about 279 nm.
v of idoxuridine RS in the same solvent mixture.
B. In the Assay, the chromatogram obtained with the reference
Apply to the plate 5 µl of each solution. After development, solution (a) shows a peak that corresponds to the peak due to
dry the plate in a current of cold air and repeat the idoxuridine in the chromatogram obtained with the test
development. After removal of the plate following the second solution.
601
IMIPENEM IP 2007
Tests Description. A white to almost white or pale yellow powder.
Other tests. Comply with the tests stated under Eye Drops. Identification
Test solution. Add 2 ml of a 10 per cent v/v solution of ethanol Compare the spectrum with that obtained with imipenem
(95 per cent) to 15.0 ml of a solution prepared by diluting an monohydrate RS or with the reference spectrum of imipenem
accurately measured volume of the eye drops with water if monohydrate.
necessary to give a final concentration of 0.1 per cent w/v of
Idoxuridine (solution A) and dilute to 20.0 ml with water. Tests
Reference solution (a). Shake 0.1 g of idoxuridine RS with 50 Appearance of solution. A 1.0 per cent w/v solution in
ml of water until dissolved and then dilute to 100.0 ml with phosphate buffer pH 7.0 is not more opalescent than
water. To 15.0 ml of this solution add 2.0 ml of a solution opalescence standard OS2 (2.4.1) and not more intensely
prepared by diluting 10 ml of a 1.2 per cent w/v of coloured than reference solution BYS6 (2.4.1).
sulphathiazole (internal standard) in ethanol (95 per cent) to
100 ml with water (solution B), and dilute to 20.0 ml with water. pH (2.4.24). 4.5 to 7.0 determined in 0.5 per cent w/v solution in
water.
Reference solution (b). Add 2.0 ml of solution B to 15.0 ml of
solution A and dilute to 20.0 ml with water. Specific optical rotation (2.4.22). + 84.0º to + 89.0º, determined
in a 0.5 per cent w/v solution in phosphate buffer pH 7.
– a stainless steel column 30 cm x 4 mm, packed with Related substances. Determine by liquid chromatography
octadecylsilyl silica gel (10 µm), (2.4.14).
– mobile phase: a mixture of 87 volumes of water and 13 NOTE –Prepare the solutions immediately before use.
volumes of methanol,
– flow rate. 1.7 ml per minute, Solvent mixture. A solution prepared by dissolving 1.36 g of
– spectrophotometer set at 254 nm, potassium dihydrogen phosphate in 1000 ml of water, adjust
– a 20 µl loop injector. to pH 6.8 with 10 per cent w/v of potassium hydroxide and
filter.
Calculate the content of C9H11IN2O5 in the eye drops.
Test solution. Dissolve about 50 mg of the substance under
Storage. Store at a temperature not exceeding 30°. The eye
drops should not be allowed to freeze.
Labelling. The label states that the eye drops should not be Reference solution (a). Heat 20 ml of the test solution at 60º
used for continuous periods of treatment exceeding 21 days. for 5 minutes previously adjusted to pH 10.0 with 10 per cent
w/v of sodium hydroxide.
imipenem monohydrate RS in the solvent mixture.
Imipenem Chromatographic system
COOH octadecylsilane bonded to porous silica (5 µm) (such as
O ACE, AQ),
H3C N – mobile phase: A. methanol,
S , H2O
H N NH B. a buffer solution prepared by dissolving 6.25 g
HO H H H of dipotassium hydrogen phosphate and 2.5 g of
potassium dihydrogen phosphate in 900 ml of water.
C12H17N3O4S,H2O Mol. Wt. 317.4 Adjust to pH 7.0 with orthophosphoric acid or 10 per
cent w/v of potassium hydroxide, dilute to 1000 ml with
Imipenem is (5R,6S)-6-[(R)-1-hydroxyethyl]-3-[[2-[ water and filter,
(iminomethyl)amino]ethyl]sulphanyl]-7-oxo-1- – flow rate.1.2 ml per minute,
azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid. – a linear gradient programme using the conditions given
Imipenem contains not less than 98.0 per cent and not more below,
than 101.0 per cent of C12H 17N 3O 4S, calculated on the – spectrophotometer set at 210 nm and 300 nm,
anhydrous basis. – a 20 µl loop injector.
602
IP 2007 IMIPENEM AND CILASTATIN INJECTION
Time Mobile phase A Mobile phase B – flow rate. 1.5 ml per minute,
( in min) (per cent v/v) (per cent v/v) – spectrophotometer set at 300 nm,
0 0 100 – a 10 µl loop injector.
13 0 100 Inject the reference solution. The test is not valid unless the
60 20 80 column efficiency is not less than 600 theoretical plates, and
the relative standard deviation for replicate injections is not
62 0 100 more than 2.0 per cent.
Inject reference solution (a) at 300 nm. The relative retention
time between the peak due to theinamycin and imipenem is
about 0.76. The test is not valid unless the resolution between Calculate the content of C12H17N3O4S.
the peak due to theinamycin and imipenem is at least 4.0. The
Imipenem intended for use in the manufacture of parenteral
column efficiency is not less than 3000 theoretical plates, and
preparations without a further appropriate procedure for
the removal of bacterial endotoxins complies with the
Inject reference solution (b) at 210 nm. The test is not vaild following additional requirement.
unless the relative standard deviation for replicate injections
Bacterial Endotoxins (2.2.3). Not more than 0.17 Endotoxin
is not more than 2.0 per cent.
unit per mg of imipenem.
Inject the test solution and reference solution (b). Continue
the chromatography for twice the retention time of the principal Imipenem intended for use in the manufacture of parenteral
peak. In the chromatogram obtained with the test solution, preparations without a further appropriate sterilisation
the area of any peak, other than that of the principal peak and procedure complies with the following additional
any peak corresponding to thienamycin, is not more than 0.7 requirement.
times the area of the principal peak in the chromatogram Sterility (2.2.11). Complies with the test for sterility.
obtained with reference solution (b) (0.7 per cent); the sum of
the areas of all the peaks, other than that of the principal peak Storage. Store in an airtight container in a refrigerator (2º to
and any peak corresponding to thienamycin, is not more than 8º).
the area of the principal peak in the chromatogram obtained Labelling. The label states, where applicable, that the
with reference solution (b) (1 per cent). Ignore any peak with substance is sterile and is free from bacterial endotoxins.
an area less than 0.1 times the area of the principal peak in the
on 1.0 g. Imipenem and Cilastatin Injection
Water (2.3.43). 5.0 per cent to 8.0 per cent, determined on Imipenem and Cilastatin Injection is a sterile mixture of
0.2 g. Imipenem, Cilastatin Sodium, and Sodium Bicarbonate.
Keep the solutions in an ice-bath and use within 8 hours of sealed container in the requisite amount of sterile Water for
preparation. Injections, immediately before use.
Test solution. Dissolve about 40 mg of the substance under The constituted solution complies with the requirements for
examination in 100 ml of the mobile phase. Clarity of solution and Particulate matter stated under
Reference solution. A 0.04 per cent w/v solution of imipenem Parenteral Preparations (Injections).
monohydrate RS in the mobile phase. Storage. The constituted solution should be used immediately
Chromatographic system after preparation but, in any case, within the period
– a stainless steel column 30 cm × 3.9 mm, packed with recommended by the manufacturer.
octadecylsilane bonded to porous silica (10 µm) (such Imipenem and Cilastatin Injection contains not less than 90.0
as Bondapak C18), per cent and not more than 115.0 per cent of the stated amounts
– mobile phase: a solution prepared by dissolving 54 mg of imipenem, C12H17N3O4S and cilastatin, C16H26N2O5S.
of monobasic potassium phosphate in 360 ml of water,
adjusted to pH 6.8 with 0.5 M sodium hydroxide or 0.5 The contents of the sealed container comply with the
M orthophosphoric acid, dilute to 400 ml with water requirements stated under Parenteral Preparations
and filter, (Powders for Injection) and with the following requirements.
603
IMIPRAMINE HYDROCHLORIDE IP 2007
Identification Storage. Store protected from moisture, in a single dose or

multiple dose container.
with the test solution corresponds to the peak in the Labelling. The label states that the constituted solution
chromatogram obtained with the reference solution. should be solubilized in a suitable parenteral fluid prior to
intravenous infusion.
Tests
pH (2.4.24). 6.5 to 8.5, when constituted as directed on the
label. Imipramine Hydrochloride
Unit per mg of imipenem and not more than 0.17 Endotoxin
Unit per mg of cilastatin.
N ,HCl
CH3
Loss on drying (2.4.19). Not more than 3.5 per cent, determined N
on 0.1 g, by drying in an oven at 60º for 3 hours at a pressure CH3
C19H24N2,HCl Mol. Wt. 316.9
Imipramine Hydrochloride is 10,11-dihydro-5H-
NOTE – Prepare the following solutions immediately before dibenz[b,f]azepine-5-(dimethylaminopropyl) hydrochloride.
use.
Imipramine Hydrochloride contains not less than 98.5 per cent
Test solution. Determine the weight of the contents of 10
and not more than 101.0 per cent of C19H24N2,HCl, calculated
on the dried basis.
of the 10 containers containing about 50 mg of Imipenem
dissolve in buffer solution pH 6.8 and dilute to 100.0 ml with Description. A white or slightly yellow, crystalline powder;
the same solution. almost odourless.
Reference solution (a). A 0.05 per cent w/v solution of Identification
imipenem monohydrate RS in buffer solution pH 6.8.
Tests B, C and D may be omitted if tests A and E are carried
cilastatin ammonium RS in buffer solution pH 6.8.
out.
– a stainless steel column 25 cm x 4.6 mm, packed with Compare the spectrum with that obtained with imipramine
octadecylsilane bonded to porous silica (5 µm), hydrochloride RS or with the reference spectrum of imipramine
– column temperature 50º, hydrochloride.
– mobile phase: dissolve 2.0 g of sodium 1-
hexanesulphonate in 800 ml of buffer solution pH 6.8, B. When examined in the range 230 nm to 360 nm (2.4.7), a
adjust the pH to 6.8 with 0.5 M sodium hydroxide or 0.5 0.002 per cent w/v solution in 0.01 M hydrochloric acid shows
M phosphoric acid, dilute to 1000 ml with buffer solution an absorption maximum only at about 250 nm and a shoulder
pH 6.8 and filter, at about 270 nm; absorbance at about 250 nm, about 0.52.
– flow rate. 2 ml per minute, C. Dissolve 5 mg in 2 ml of nitric acid; an intense blue colour
– spectrophotometer set at 254 nm, is produced.
D. Dissolve 50 mg in 3 ml of water and add 1 drop of a 2.5 per
Inject reference solution (a). The test is not valid unless the cent w/v solution of quinhydrone in methanol; no red colour
tailing factor is not more than 1.5, the column efficiency in not is produced within 15 minutes.
less than 600 theoretical plates, and the relative standard
E. 20 mg gives reaction A of chlorides (2.3.1).
Inject alternately the test solution and reference solutions (a) Tests
and (b). Appearance of solution. Triturate with a glass rod 3.0 g with
Calculate the contents of C12H17N3O4S and C16H26N2O5S in the 20 ml of carbon dioxide-free water and dilute to 30 ml with the
injection. same solvent (solution A). Solution A is clear (2.4.1).
604
IP 2007 IMIPRAMINE TABLETS
Immediately after preparation dilute the solution with an equal Identification

volume of water. The resulting solution is not more intensely
coloured than reference solution BYS6 (2.4.1). Triturate a quantity of the powdered tablets containing about
0.25 g of Imipramine Hydrochloride with 10 ml of chloroform,
pH (2.4.24). 3.6 to 5.0, determined in solution A immediately filter, evaporate the filtrate to low bulk, add ether until a turbidity
after preparation. is produced, and allow to stand. The precipitate, after
Related substances. Determine by thin-layer chromatography recrystallisation from acetone, melts at about 172° (2.4.21),
(2.4.17), coating the plate with silica gel G. and complies with the following tests.
Mobile phase. A mixture of 55 volumes of ethyl acetate, 35 A. Dissolve 5 mg in 2 ml of nitric acid; an intense blue colour
volumes of glacial acetic acid, 5 volumes of hydrochloric is produced.
acid and 5 volumes of water. B. Dissolve 50 mg in 3 ml of water and add 1 drop of a 2.5 per
Prepare the following solutions immediately before use. cent w/v solution of quinhydrone in methanol; no red colour
Test solution. Dissolve 0.25 g of the substance under is produced within 15 minutes.
examination in 10 ml of methanol. C. 20 mg gives reaction A of chlorides (2.3.1).
Tests
Reference solution (b). A 0.005 per cent w/v solution of Related substances. Determine by thin-layer chromatography
iminodibenzyl RS in methanol. (2.4.17), coating the plate with silica gel G.
Apply to the plate 10 µl of each solution. After development, Mobile phase. A mixture of 55 volumes of ethyl acetate, 35
remove the plate, allow the solvent to evaporate for 5 minutes, volumes of glacial acetic acid, 5 volumes of hydrochloric
spray with a 0.5 per cent w/v solution of potassium dichromate acid and 5 volumes of water.
in sulphuric acid (20 per cent) and examine immediately. In Prepare the following solutions immediately before use.
the chromatogram obtained with the test solution any spot
corresponding to iminodibenzyl is not more intense than the Test solution. Shake a quantity of the powdered tablets
spot in the chromatogram obtained with reference solution containing 0.2 g of Imipramine Hydrochloride with three
(b) and any other secondary spot is not more intense than the quantities, each of 10 ml, of chloroform, filter the combined
spot in the chromatogram obtained with reference solution chloroform extracts, evaporate to dryness and dissolve the
(a). residue in 10 ml of methanol.
Heavy metals (2.3.13). 2.0 g complies with the limit test for Reference solution (a). Dilute 3 volumes of the test solution
heavy metals, Method B (10 ppm). to 1000 volumes with methanol.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Reference solution (b). A 0.006 per cent w/v solution of
iminodibenzyl RS in methanol.
on 1.0 g by drying in an oven at 105°. Apply to the plate 10 µl of each solution. After development,
remove the plate, allow the solvent to evaporate for 5 minutes,
Assay. Weigh accurately about 0.5 g, dissolve in 50 ml of spray with a 0.5 per cent w/v solution of potassium dichromate
chloroform, add 10 ml of mercuric acetate solution and titrate in sulphuric acid (20 per cent) and examine immediately. In
with 0.1 M perchloric acid, using 0.5 ml of metanil yellow the chromatogram obtained with the test solution any spot
solution as indicator. Carry out a blank titration. corresponding to iminodibenzyl is not more intense than the
1 ml of 0.1 M perchloric acid is equivalent to 0.03169 g of spot in the chromatogram obtained with reference solution
C19H24N2,HCl. (b) and any other secondary spot is not more intense than the
Storage. Store protected from light. spot in the chromatogram obtained with reference solution
(a).
Uniformity of content. (For tablets containing 10 mg or less).
Imipramine Tablets Comply with the test stated under Tablets.

Powder one tablet, shake with 25 ml of 0.1 M hydrochloric
Imipramine HydrochlorideTablets acid for 30 minutes, add sufficient 0.1 M hydrochloric acid to
Imipramine Tablets contain not less than 92.5 per cent and not produce 100.0 ml and filter. Dilute 10.0 ml of the filtrate to 50.0
more than 107.5 per cent of the stated amount of imipramine ml with 0.1 M hydrochloric acid and measure the absorbance
hydrochloride, C19H24N2,HCl. The tablets are coated. of the resulting solution at the maximum at about 250 nm (2.4.7).
605
INDINAVIR SULPHATE IP 2007
Calculate the content of C19H24N2,HCl in the tablet taking 264 Specific optical rotation (2.4.22). +122° to +129°, determined
as the specific absorbance at 250 nm. at about 365 nm in a 1.0 per cent w/v solution in water,
Other tests. Comply with the tests stated under Tablets. calculated on the anhydrous and ethanol-free basis.
(2.4.14).
quantity of the powder containing about 25 mg of Imipramine
Hydrochloride, shake with 75 ml of 0.1 M hydrochloric acid Test solution. Dissolve 0.1 g of the substance under
for 30 minutes, dilute to 100.0 ml with the same solvent and examination in 100 ml of the mobile phase.
filter through a sintered-glass filter. Dilute 10.0 ml to 100.0 ml Reference solution (a). A 0.0001 per cent w/v solution of
with 0.1 M hydrochloric acid and measure the absorbance of indinavir RS in the mobile phase.
the resulting solution at the maximum at about 250 nm (2.4.7).
Reference solution (b). Transfer 15 mg of indinavir RS to a
Calculate the content of C19H24N2, HCl taking 264 as the
50-ml volumetric flask and add 0.1 ml of 5 M hydrochloric
acid. Keep this solution for about 1 hour at room temperature
and make up to volume with the mobile phase.
Indinavir Sulphate – a stainless steel column 20 cm x 4.6 mm, packed with
base deactivated octylsilyl silica gel (5 µm),
– column temperature. 40°,
40 volumes of acetonitrile and 60 volumes of a solution
N N OH OH containing 0.37 per cent w/v of sodium citrate and
H
N N ,H2SO4 0.16 per cent w/v of citric acid, the pH of which has
been adjusted to 5.0 with 1 M sodium hydroxide or 1 M
C(CH3 )3 O phosphoric acid,
O N
H – flow rate. 1 ml per minute,
C36H47N5O4 , H2SO4 Mol. Wt. 711.9
Indinavir Sulphate is [1(1S,2R),5(S)]-2,3,5-trideoxy-N-(2,3- resolution between the peaks due to indinavir and any impurity
dihydro-2-hydroxy-1H-inden-1-yl)-5-[2-[{1,1- at a relative retention time of about 1.4 is not less than 2.
dimethylethyl)amino}carbonyl]-4-(3-pyridinylmethyl)-1-
piperazinyl]-2-(phenylmethyl)-D-erythro-pentonamide Inject reference solution (a). The test is not valid unless the
sulphate. capacity factor for indinavir peak is not less than 2.0, the
Indinavir Sulphate contains not less than 98.5 per cent and deviation for replicate injections is not more than 5 per cent.
not more than 101.5 per cent of C36H47N5O4,H2SO4, calculated
on the anhydrous and ethanol-free basis. Inject the test solution. Calculate the content of each impurity
in the chromatogram obtained with the test solution by
Description. A white or almost white powder; hygroscopic. comparing with the area of the principal peak obtained with
reference solution (a). The content of any individual impurity
Identification is not more than 0.1 per cent and the sum of all impurities is
A. When examined in the range 200 nm to 300 nm (2.4.7), a not more than 0.5 per cent.
0.005 per cent w/v solution in water shows an absorption Monoethyl sulphate content. Determine by liquid
maximum at about 260nm. chromatography (2.4.14).
B. In the Assay, the principal peak in the chromatogram Prepare the following solutions freshly.
obtained with the test solution corresponds to the peak due Test solution. Dissolve about 0.125 g of the substance under
to indinavir in the chromatogram obtained with the reference examination in 25 ml of water and filter through a fine porosity
solution. membrane.
Tests Reference solution (a). Weigh and transfer about 65 mg of
potassium monoethyl sulphate to a 50-ml volumetric flask,
pH (2.4.24). 2.8 to 3.2, determined in a 1.0 per cent w/v solution dissolve and dilute to volume with water. Dilute 5 ml of this
in carbon dioxide-free water. solution to 50 ml with water.
606
IP 2007 INDINAVIR CAPSULES
Reference solution (b). Dilute 5 ml of the test solution to 50 ml Weigh accurately about 0.5 g, dissolve in 50 ml of
with water. Further dilute 25 ml of this solution to 100 ml with dimethylformamide and titrate with 0.1 M sodium hydroxide,
water and filter through a fine porosity membrane. determining the end-point potentiometrically (2.4.25). Carry
Chromatographic system out a blank titration.
– a stainless steel column 25 cm x 4.6 mm, packed with 1 ml of 0.1 M sodium hydroxide is equivalent to 0.0480 g of
rigid, spherical styrene-divinylbenzenecopolymer, 5 to sulphate.
10 µm (such as Metrosep A supp 3, 6.1005.320), Heavy metals (2.3.13). 1.0 g complies with the limit test for
– mobile phase: a mixture of 10 volumes of acetone and 90 heavy metals, Method B (20 ppm).
volumes of a buffer prepared by dissolving 0.25 g each
of sodium carbonate and sodium bicarbonate in 1000 Sulphated ash (2.3.18). Not more than 0.1 per cent.
ml of water, Water (2.3.43). Not more than 1.5 per cent, determined on 0.2 g.
– flow rate. 0.5 ml per minute, Assay. Determine by liquid chromatography (2.4.14).
– a conductivity detector,
– a 20 µl loop injector. Test solution. Weigh accurately about 60 mg of the substance
under examination in 100.0 ml of the mobile phase.
Reference solution. A 0.06 per cent w/v solution of indinavir
relative standard deviation of replicate injections is not more
RS in the mobile phase.
than 2.0 per cent and the tailing factor for monoethyl sulphate
anion peak is not more than 2.0. Chromatographic system
Inject the test solution. Calculate the content of monoethyl
base deactivated octylsilyl silica gel (5 µm),
sulphate present from the declared content of monoethyl
sulphate present in potassium monoethyl sulphate.
– mobile phase: a filtered and degassed mixture of 45
1 mg of potassium monoethyl sulphate corresponds to 0.77 volumes of acetonitrile and 55 volumes of a buffer
mg of monoethyl sulphate. The content of monoethyl sulphate prepared by dissolving 3 g of phosphoric acid and 1.7
is not more than 0.05 per cent w/w. ml of dibutylamine in 900 ml of water, adjusting the pH
Ethanol. 5.0 to 8.0 per cent, calculated on the anhydrous basis, to 6.5 with 1 M sodium hydroxide and making up the
determined by the following method. volume to 1000.0 ml with water,
Determine by gas chromatography (2.4.13). – spectrophotometer set at 260 nm,
Test solution. Add 5 ml of a 1.5 per cent w/v solution of n- – a 10 µl loop injector.
propanol (internal standard) to 1.0 g of the substance under Inject the reference solution. The test is not valid unless the
examination in a 25-ml volumetric flask and dilute to volume column efficiency determined from the indinavir peak is not
with water. less than 4000 theoretical plates, the tailing factor is not more
Reference solution. A 0.1 per cent w/v solution of ethanol. than 2.0 and the relative standard deviation for replicate
Transfer 5 ml of this solution and 5 ml of the internal standard injections is not more than 2.0 per cent.
to a 25-ml volumetric flask and make up to volume with water. Inject alternately the test solution and the reference solution.
Chromatographic system Calculate the content of C36H47N5O4, H2SO4.
– a stainless steel column 1.8 m x 2 mm, packed with
1 mg of indinavir corresponds to 1.16 mg of indinavir sulphate.
ethylvinyl benzene divinyl benzene copolymer, mesh
size 80/100, Storage. Store protected from light.
– temperature:
inlet port. 180°,
– flame ionisation detector, Indinavir Capsules
– flow rate. 20 ml per minute of the carrier gas (nitrogen). Indinavir Sulphate Capsules
Separately inject 10 µl of the test solution and the reference Indinavir Capsules contain not less than 90.0 per cent and not
solution. Calculate the ethanol content by comparing the ratio more than 110.0 per cent of the stated amount of indinavir,
of the area of the peak corresponding to ethanol and the area C36H47N5O4.
of the internal standard peak in the chromatogram obtained
with the test solution with that of the reference solution. Identification
Sulphate.13.2 to 14.4 per cent w/w, calculated on the anhydrous A. Shake a quantity of the contents of the capsules containing
and ethanol-free basis, determined by the following method. 0.1 g of Indinavir Sulphate with 80 ml of water, dilute to 100 ml
607
INDINAVIR CAPSULES IP 2007
with water and filter. Dilute 5 ml of the filtrate to 100 ml with Inject reference solution (b). The test is not valid unless the
water. When examined in the range 200 nm to 300 nm (2.4.7), column efficiency determined from the indinavir peak is not
the resulting solution shows an absorption maximum at about less than 10,000 theoretical plates, the tailing factor is not
260 nm. more than 1.5 and the resolution between indinavir and
B. In the Assay, the principal peak in the chromatogram indinavir 4-epimer peaks is not less than 1.5.
obtained with the test solution corresponds to the peak due Inject reference solution (a) and the test solution. Examine the
to indinavir in the chromatogram obtained with the reference chromatogram obtained with reference solution (a) for any
solution. extraneous peaks and ignore the corresponding peaks
observed in the chromatogram obtained with the test solution.
Tests
Related substances. Determine by liquid chromatography with the test solution should not be more than 1.0 per cent
(2.4.14). and the sum of the areas of all the secondary peaks should
Test solution. Mix well the contents of 20 capsules and shake not be more than 2.5 per cent when calculated by percentage
a quantity of the mixed contents containing about 50 mg of area normalisation.
indinavir with about 60 ml of a solution prepared by mixing 40 Dissolution (2.5.2).
volumes of acetonitrile and 60 volumes of 0.05 M dipotassium
hydrogen phosphate, the pH of which is adjusted to 7.5 with Apparatus. No 1
dilute phosphoric acid (solution A) in a 100-ml volumetric Medium. 900 ml of a buffer prepared by dissolving 21 g of
flask, mix with the aid of ultrasound for 10 minutes, dilute to citric acid in 880 ml of water, adjusting the pH to 3.8 with a 50
volume with solution A and filter. per cent w/v solution of sodium hydroxide and making up to
1000 ml with water
Reference solution (a). Weigh accurately a quantity of
indinavir sulphate RS containing about 50 mg of indinavir in Speed and time. 50 rpm and 30 minutes. Use sinkers.
a 100-ml volumetric flask, dissolve and dilute to volume with Withdraw a suitable volume of the medium and filter through
solution A. Dilute 1 ml of this solution to 100 ml with solution a membrane filter disc with an average pore diameter not greater
A. than 1.0 µm. Reject the first few ml of the filtrate and dilute a
Reference solution (b). Dissolve a quantity of indinavir suitable volume of the filtrate if necessary, with the same
sulphate RS containing about 50 mg of indinavir and 5 mg of solvent. Measure the absorbance of the resulting solution at
indinavir 4-epimer RS and dilute to 100 ml with solution A. the maximum at about 260 nm (2.4.7). Calculate the content of
indinavir, C36H47N5O4, in the medium from the absorbance
obtained from a solution of known concentration of indinavir
sulphate RS in the same solvent.
– mobile phase: filtered and degassed mixtures of D. Not less than 75 per cent of the stated amount of
acetonitrile and 0.05 M dipotassium hydrogen C36H47N5O4.
phosphate, adjusting the pH of the solution to 7.5 with Other tests. Comply with the tests stated under Capsules.
dilute phosphoric acid,
– flow rate. 1 ml per minute, Assay. Determine by liquid chromatography (2.4.14).
– a linear gradient programme using the conditions given Test solution. Mix well the contents of 20 capsules and shake
below, a quantity of the mixed contents containing about 200 mg of
– spectrophotometer set at 260 nm, indinavir with about 60 ml of the mobile phase, mix with the aid
– a 20 µl loop injector. of ultrasound for 10 minutes, dilute to 100.0 ml with the mobile
Time Phosphate buffer (pH 7.5) Acetonitrile phase, mix and filter. Dilute 10.0 ml of the filtrate to 100.0 ml
(in min) (per cent v/v) (per cent v/v) with the mobile phase.
0 80 20 Reference solution. Weigh accurately a quantity of indinavir
3 80 20 sulphate RS containing about 50 mg of indinavir in a 50-ml
5 65 35 volumetric flask, dissolve and dilute to volume with the mobile
11 65 35 phase. Dilute 10.0 ml of this solution to 50.0 ml with the mobile
17 30 70 phase.
20 30 70 Chromatographic system
21 80 20 – a stainless steel column 25 cm x 4.6 mm, packed with
25 80 20 base deactivated octylsilyl silica gel (5 µm),
608
IP 2007 INDOMETHACIN CAPSULES
– mobile phase: a filtered and degassed mixture of methanol and 10 volumes of 1 M hydrochloric acid shows an
45 volumes of acetonitrile and 60 volumes of 0.05 M absorption maximum only at about 320 nm; absorbance at
dipotassium hydrogen phosphate, with the pH adjusted about 320 nm, about 0.45.
to 7.5 with dilute phosphoric acid, C. Dissolve 0.1 g in 10 ml of ethanol (95 per cent), heating
– flow rate. 1 ml per minute, gently if necessary. To 0.1 ml add 2 ml of a freshly prepared
– spectrophotometer set at 260 nm, mixture of 1 volume of a 25 per cent w/v solution of
– a 20 µl loop injector. hydroxylamine hydrochloride and 3 volumes of 2 M sodium
Inject the reference solution. The test is not valid unless the hydroxide. Add 2 ml of 2 M hydrochloric acid and 1 ml of
column efficiency determined from the indinavir peak is not ferric chloride solution and mix; a violet-pink colour develops.
less than 6000 theoretical plates, the tailing factor is not more
than 1.5 and the relative standard deviation for replicate Tests
Inject alternately the test solution and the reference solution. (2.4.17), coating the plate with a suspension of silica gel
Calculate the content of C36H47N5O4 in the capsules. HF254 in a 4.68 per cent w/v solution of sodium dihydrogen
phosphate.
exceeding 30° Mobile phase. A mixture of 70 volumes of ether and 30 volumes
of light petroleum (50° to 70°).
Indomethacin Test solution. Dissolve 0.2 g of the substance under
Indometacin
CH3 substance under examination in methanol.
O
N COOH dry the plate in air and examine in ultraviolet light at 254 nm.
Cl test solution is not more intense than the spot in the
OCH3 Heavy metals (2.3.13). 1.0 g complies with the limit test for
C19H16ClNO4 Mol. Wt. 357.8
Indomethacin is 1-(4-chlorobenzoyl)-5-methoxy-2-
methylindol-3-yl acetic acid. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Indomethacin contains not less than 98.0 per cent and not
more than 101.0 per cent of C19H16ClNO4, calculated on the Assay. Weigh accurately about 0.45 g, dissolve in 75 ml of
dried basis. acetone and titrate under nitrogen with carbonate-free 0.1 M
sodium hydroxide using 0.2 ml of phenolphthalein solution
Description. A white to pale yellow, crystalline powder; as indicator. Carry out a blank titration.
Identification C19H16ClNO4.
Test A may be omitted if tests B and C are carried out. Tests B Storage. Store protected from light.
Compare the spectrum with that obtained with indomethacin
RS or with the reference spectrum of indomethacin. Examine Indomethacin Capsules
the substances in the solid state without recrystallisation. Indomethacin Capsules contain not less than 90.0 per cent
B. When examined in the range 230 nm to 360 nm (2.4.7), a and not more than 110.0 per cent of the stated amount of
0.0025 per cent w/v solution in a mixture of 90 volumes of indomethacin, C19H16ClNO4.
609
INDOMETHACIN SUPPOSITORIES IP 2007
Identification medium from the absorbance obtained from a solution of known

concentration of indomethacin RS in the same medium.
A. Shake a quantity of the contents of the capsules containing
0.1 g of Indomethacin with 5 ml of chloroform, filter and D. Not less than 80 per cent of the stated amount of
evaporate the filtrate to dryness. Dry the residue at 60° at a C19H16ClNO4.
pressure not exceeding 0.7 kPa for 1 hour. The residue complies Other tests. Comply with the tests stated under Capsules.
Assay. Weigh accurately a quantity of the mixed contents of
Determine by infrared absorption spectrophotometry (2.4.6). 20 capsules containing about 50 mg of Indomethacin, add
Compare the spectrum with that obtained with indomethacin 10 ml of water and allow to stand for 10 minutes, with
RS or with the reference spectrum of indomethacin. occasional swirling. Add 75 ml of methanol, shake well, add
sufficient methanol to produce 100.0 ml and filter if necessary.
To 5.0 ml of the filtrate add sufficient of a mixture of equal
volumes of methanol and phosphate buffer pH 7.2 to produce
100.0 ml. Measure the absorbance of the resulting solution at
C. Mix a quantity of the contents of the capsules containing the maximum at about 320 nm (2.4.7). Calculate the content of
25 mg of Indomethacin with 2 ml of water and add 2 ml of 2 M C19H16ClNO4 taking 193 as the specific absorbance at 320 nm.
sodium hydroxide; a bright yellow colour is produced which Storage. Store protected from moisture.
fades rapidly.
Tests
Indomethacin Suppositories
(2.4.17), coating the plate with a suspension of silica gel Indomethacin Suppositories contain Indomethacin in a suitable
HF254 in a 4.68 per cent w/v solution of sodium dihydrogen suppository basis.
phosphate. Indomethacin Suppositories contain not less than 90.0 per
Mobile phase. A mixture of 70 volumes of ether and 30 volumes cent and not more than 110.0 per cent of the stated amount of
of light petroleum (50° to 70°). indomethacin, C19H16ClNO4.
Prepare the following solutions immediately before use. Identification

Test solution. Shake a quantity of the contents of the capsules A. Dissolve a quantity of the powdered suppositories
containing 0.1 g of Indomethacin with 5 ml of chloroform, containing 0.1 g of Indomethacin as completely as possible in
filter and use the filtrate. 50 ml of hot water, filter, wash the residue with hot water and
Reference solution. Dilute 1 volume of the test solution to 200 allow to dry in air. Dissolve the residue in 5 ml of chloroform
volumes with chloroform. and evaporate to dryness. The residue complies with the
following test.
dry the plate in air and examine in ultraviolet light at 254 nm. Determine by infrared absorption spectrophotometry (2.4.6).
Any secondary spot in the chromatogram obtained with the Compare the spectrum with that obtained with indomethacin
test solution is not more intense than the spot in the RS or with the reference spectrum of indomethacin.
chromatogram obtained with the reference solution. B. Shake a quantity containing 25 mg of Indomethacin with 5
Dissolution (2.5.2). ml of water until the base dissolves; a white suspension is
produced. Add 2 ml of 2 M sodium hydroxide; a bright yellow
Apparatus. No 1 colour is produced which fades rapidly.
Medium. 750 ml of a freshly prepared mixture of 1 volume of
phosphate buffer pH 7.2 and 4 volumes of water Tests
Withdraw a suitable volume of the medium and filter promptly (2.4.14).
Dilute the filtrate suitably with the medium and measure the Test solution (a). Powder or cut into small pieces a suitable
absorbance of the resulting solution at the maximum at about number of suppositories, dissolve a quantity containing 0.1 g
320 nm (2.4.7). Calculate the content of C19H16ClNO4 in the of Indomethacin in sufficient methanol to produce 50 ml.
610
IP 2007 INSULIN
Reference solution (a). Dilute 3 volumes of test solution (a) Insulin

to 100 volumes with the mobile phase.
Crystalline Insulin
octadecylsilyl silica gel (10 µm)(such as Bondapack C18), Gly
– mobile phase: a mixture of 60 volumes of methanol and Ile
40 volumes of 0.2 per cent v/v solution of phosphoric Val
acid, Glu
Gln-Cys-Cys-Ala-Ser-Val-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn-Tyr-Cys-Asn
– spectrophotometer set at 320 nm, His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-Cys
– a 20 µl loop injector. Gln Gly
Asn Glu
The sum of the areas of any secondary peaks that elute before Val Arg
the principal peak in the chromatogram obtained with test Ala-Lys-Pro-Thr-Tyr-Phe-Phe-Gly
Phe
solution (a) is not greater than the area of the peak in the
C256H381N65O76S6 (porcine) Mol. Wt. 5777.6
Repeat the procedure but using the following freshly prepared
solutions and a detection wavelength of about 240 nm.
Gly
Test solution (b). Dilute 10 volumes of test solution (a) to 20
Ile
volumes with the mobile phase. Val
Reference solution (b). A solution containing 0.001 per cent Glu
w/v of 4-chlorobenzoic acid in the mobile phase. Gln-Cys-Cys-Thr-Ser-Ile-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn-Tyr-Cys-Asn
In the chromatogram obtained with test solution (b) the sum His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-Cys
of the areas of any secondary peaks that elute before the Gln Gly
principal peak, other than those determined in test solution Asn Glu
(a), is not greater than the area of the peak in the chromatogram Val Arg
obtained with reference solution (b). Phe Ala-Lys-Pro-Thr-Tyr-Phe-Phe-Gly
The column efficiency, determined using the principal peak in

the chromatogram obtained with reference solution (a), should C254H377N65O75S6 (bovine) Mol. Wt. 5733.5
be not less than 7500 theoretical plates per metre.
Insulin is the specific natural antidiabetic principle obtained
Disintegration (2.5.1). Use a weighed suppository and from the pancreas of either the pig or the ox and purified.
phosphate buffer pH 6.8 in place of water and operate the
Insulin contains not less than 26.5 Units per mg of porcine
apparatus for 90 minutes. At the end of this period remove the
insulin, C256H381N65O76S6, or of bovine insulin, C254H377N65O75S6,
suppository, dry with filter paper and weigh. Repeat the
as appropriate, calculated on the dried basis.
operation with two further weighed suppositories. Not less
than 75 per cent of each suppository is dissolved. Description. A white or almost white powder.
Other tests. Comply with the tests stated under Suppositories.
Identification
Assay. Weigh 10 suppositories and powder or cut into small
pieces. Weigh accurately a quantity of the powder or small A. In the Assay, the principal peak due to insulin in the
pieces containing about 0.1 g of Indomethacin, add 50 ml of chromatogram obtained with the test solution corresponds to
methanol, shake until the dispersion is complete and, if the peak in the chromatogram obtained with the appropriate
necessary, filter. To 2.0 ml add sufficient of a mixture of equal reference solution.
volumes of methanol and phosphate buffer pH 7.2 to produce B. Complies with the test for peptide mapping (2.3.47).
100.0 ml. Measure the absorbance of the resulting solution at
the maximum at about 320 nm (2.4.7). Calculate the content of Tests
C19H16ClNO4 taking 193 as the specific absorbance at 320 nm.
Storage. Store protected from moisture at a temperature not solution in 0.01 M hydrochloric acid at the maximum at about
exceeding 30°. 276 nm, 0.48 to 0.56.
611
HUMAN INSULIN IP 2007
Other tests. Complies with the tests for Impurities with purification of insulin obtained from the pancreas of the pig
molecular masses greater than that of insulin, Related proteins or by a method based on recombinant DNA (rDNA).
and Total zinc stated under Insulin Preparations. Human insulin contains not less than 27.5 Units per mg of
Sulphated ash (2.3.18). Not more than 2.5 per cent, calculated human insulin, C257H383N65O77S6, calculated on the dried basis.
on the dried basis, determined on 0.2 g.
determined on 0.2 g by drying in an oven at 60° at a pressure Identification
A. In the Assay, the principal peak due to insulin in the
Assay. Determine as described under Assay of Insulins (2.3.46). chromatogram obtained with the test solution corresponds to
Test solution. Dissolve a suitable quantity of the substance the peak in the chromatogram obtained with reference solution
under examination in 0.01 M hydrochloric acid to obtain a (b) or (c), as appropriate.
concentration of 4.0 mg per ml. B. Complies with the test for peptide mapping (2.3.47).
Insulin intended for use in the manufacture of parenteral
preparations without a further appropriate procedure for Tests
the removal of bacterial endotoxins complies with the Light absorption (2.4.7). Absorbance of a 0.05 per cent w/v
following additional requirement. solution in 0.01 M hydrochloric acid at the maximum at about
Bacterial endotoxins (2.2.3). Not more than 20 Endotoxin Units 276 nm, 0.48 to 0.56.
per mg. Total zinc. Not more than 1.0 per cent, calculated on the dried
Storage. Store protected from light, at a temperature not basis, determined by the following method. To 5 ml of a 0.5 per
exceeding –20° until released by the manufacturer. When cent w/v solution in 0.01 M hydrochloric acid add 10 ml of
thawed, it should be stored in a refrigerator (2° to 8°) and used alkaline borate buffer pH 9.0, 3 ml of zincon solution and
for the manufacture of preparations within a short period of sufficient water to produce 50 ml. Allow to stand for 1 hour
time. and measure the absorbance of the resulting solution at about
620 nm, using as the blank a solution prepared by treating 5 ml
Labelling. The label states (1) the animal source or sources of
of water instead of the substance being examined in a similar
the insulin; (2) where applicable, that the material is free from
manner. Calculate the content of zinc from the absorbance
bacterial endotoxins; (3) the storage conditions.
obtained by repeating the procedure using a suitable aliquot
NOTE — 0.0345 mg of porcine or 0.0342 mg of bovine insulin of a mixture of 4 volumes of zinc sulphate solution and
is equivalent to 1 Unit of insulin. 6 volumes of water.
Other tests. Complies with the tests for Impurities with
molecular masses greater than that of insulin, Related proteins
Human Insulin and Total zinc stated under Insulin Preparations.
Sulphated ash (2.3.18). Not more than 2.5 per cent, calculated
Gly on the dried basis, determined on 0.2 g.
Ile Loss on drying (2.4.19). Not more than 10.0 per cent,
Val
determined on 0.2 g by drying in an oven at 60° at a pressure
Glu
Gln-Cys-Cys-Thr-Ser-Ile-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn-Tyr-Cys-Asn
Assay. Determine as described under Assay of Insulins (2.3.46).
His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-Cys
Gln Gly Test solution. Dissolve a suitable quantity of the substance
Asn Glu under examination in 0.01 M hydrochloric acid to obtain a
Val Arg concentration of 4.0 mg per ml.
Phe Thr-Lys-Pro-Thr-Tyr-Phe-Phe-Gly Human Insulin intended for use in the manufacture of
C257H383N65O77S6 Mol. Wt. 5808.0 procedure for the removal of bacterial endotoxins complies
Human insulin is a 2-chain peptide having the structure of
the antidiabetic hormone produced by the human pancreas. Bacterial endotoxins (2.2.3). Not more than 10 Endotoxin Units
It is produced either by enzymatic modification and suitable per mg.
612
IP 2007 BIPHASIC INSULIN INJECTION
Storage. Store protected from light, at a temperature not 0.01 M hydrochloric acid is necessary to avoid overloading
exceeding –18° until released by the manufacturer. When the column. Dilute a suitable volume of the injection with 0.01
thawed, it should be stored in a refrigerator (2° to 8°) and used M hydrochloric acid to obtain a concentration of 40 Units per
for the manufacture of preparations within a short period of ml.
time. Storage. Store in multiple dose containers in a refrigerator (2°
Labelling. The label states (1) whether the material is produced to 8°). It should not be allowed to freeze.
by enzymatic modification of porcine insulin or by rDNA
Labelling. The label states (1) whether the preparation is
technology; (2) where applicable, that the material is free from
acidified or neutral; (2) the strength in terms of the number of
bacterial endotoxins; (3) the storage conditions.
Units per ml; (3) the animal source or sources of the insulin;
NOTE - 0.0347 mg of human insulin is equivalent to 1 Unit of (4) that the preparation should not be allowed to freeze; (5)
insulin. the storage conditions.
Insulin Injection Biphasic Insulin Injection

Neutral Insulin; Neutral Insulin Injection; Soluble Insulin; Biphasic Insulin Injection is a sterile suspension of crystals
Plain Insulin containing bovine insulin in a solution of porcine insulin.
Insulin injection is a neutral, sterile solution of bovine, porcine Biphasic Insulin Injection contains not less than 90.0 per cent
or human insulin. and not more than 110.0 per cent of the stated number of Units
of Insulin.
Insulin Injection contains not less than 90.0 per cent and not
Description. A white suspension. When examined under a
more than 110.0 per cent of the stated number of Units of
microscope, the majority of the particles appear as
Insulin.
rhombohedral crystals, with a maximum dimension of the
Description. A colourless liquid, free from turbidity and foreign crystals greater than 10 µm but rarely exceeding 40 µm.
matter; during storage, traces of a very fine sediment may be
deposited. Identification
Identification In the chromatograms obtained in the Assay the position of

the peaks due to the two insulins in the chromatogram
In the chromatograms obtained in the assay, the position of obtained with the test solution correspond to those of the
the peak due to insulin in the chromatogram obtained with the principal peaks in the chromatogram obtained with the
test solution corresponds to that of the principal peak obtained appropriate reference solution.
with the appropriate reference solution.
Tests
Tests
pH (2.4.24). 6.6 to 7.2.
pH (2.4.24). 6.8 to 7.8.
Total zinc. 26.0 µg to 37.5 µg per 100 Units of insulin, determined
Total zinc. Not more than 40 µg per 100 Units of insulin, by either of the methods stated under Insulin Preparations.
determined by either of the methods stated under Insulin
Insulin in the supernatant. 22.0 per cent to 28.0 per cent of
Preparations.
insulin in solution, determined as stated under Insulin
Test solution. Dilute a volume of the gently shaken preparation Preparations.
under examination containing 200 Units to 25.0 ml with water.
Other tests. Complies with the tests stated under Insulin
Dilute if necessary to a suitable concentration with water.
Preparations.
Preparations.
Test solution. To 10 ml of the preparation under examination
Assay. Determine as described under Assay of Insulins (2.3.46). add 40 µl of 5 M hydrochloric acid, mix well, allow to stand for
Test solution. Add 4 µl of 6 M hydrochloric acid per millilitre 1 hour to ensure solution of the sediment and dilute with 0.03
of the injection to obtain a clear solution. For a preparation M hydrochloric acid to obtain a solution containing 40 Units
containing more than 100 IU ml, an additional dilution with per ml.
613
BIPHASIC ISOPHANE INSULIN INJECTION IP 2007
Biphasic Isophane Insulin Injection It is prepared by the procedure described under Insulin
Preparations. The amount of protamine is based on the known
Biphasic isophane Insulin Injection is a sterile buffered isophane ratio and is not less than the equivalent of 0.3 mg
suspension of either porcine or human insulin, complexed and not more than the equivalent of 0.6 mg of protamine
with protamine sulphate or any other suitable protamine, in a sulphate for each 100 Units of insulin in the insulin-protamine
solution of insulin of the same species. complex. It contains Sodium Phosphate as a buffering agent.
It is prepared by the procedure described under Insulin Isophane Insulin Injection contains not less than 90.0 per
Preparations. It is produced by mixing, in defined ratios, soluble cent and not more than 110.0 per cent of the stated number of
insulin injection and isophane insulin injection. Units of Insulin.
Biphasic isophane Insulin Injection contains not less than Description. A white suspension which on standing deposits
90.0 per cent and not more than 110.0 per cent of the stated a white sediment and leaves a colourless or almost colourless
number of Units of Insulin. supernatant liquid; the sediment is readily resuspended by
Description. A white suspension which on standing deposits gently shaking. When examined under a microscope, the
a white sediment and leaves a colourless or almost colourless particles are seen to be rod-shaped crystals, the majority with
supernatant liquid; the sediment is readily resuspended by a maximum dimension greater than 1 µm but rarely exceeding
gently shaking. When examined under a microscope, the 60 µm, free from large aggregates.
particles are seen to be rod-shaped crystals, the majority with
Identification
a maximum dimension greater than 1 µm but rarely exceeding
60 µm, free from large aggregates. In the chromatograms obtained in the Assay the position of
the peak due to insulin in the chromatogram obtained with the
Identification test solution corresponds to that of the principal peak in the
In the chromatograms obtained in the Assay the position of chromatogram obtained with the appropriate reference
the peak due to insulin in the chromatogram obtained with the solution.
test solution corresponds to that of the principal peak in the Tests
chromatogram obtained with the appropriate reference
solution. pH (2.4.24). 6.9 to 7.5.
Tests Total zinc. Not more than 40 µg per 100 Units of insulin,
pH (2.4.24). 6.9 to 7.5. Preparations.
Total zinc. Not more than 40 µg per 100 Units of insulin, Other tests. Complies with the tests stated under Insulin
determined by either of the methods stated under Insulin Preparations.
Preparations.
Preparations.
add 40 µl of 5 M hydrochloric acid, mix well, allow to stand for
Assay. Determine as described under Assay of Insulins (2.3.46). 1 hour to ensure solution of the sediment and dilute with 0.03
Test solution. To 10 ml of the preparation under examination M hydrochloric acid to obtain a solution containing 40 Units
add 40 µl of 5 M hydrochloric acid, mix well, allow to stand for per ml.
1 hour to ensure solution of the sediment and dilute with 0.03 Storage. Store in multiple dose containers in a refrigerator (2°
M hydrochloric acid to obtain a solution containing 40 Units to 8°). It should not be allowed to freeze.
per ml.
Labelling. The label states (1) the strength in terms of the
Labelling. The label states the ratio of soluble insulin injection number of Units per ml; (2) the animal source or sources of the
to isophane insulin injection used in the manufacture of insulin; (3) that the preparation should not be allowed to freeze;
biphasic isophane insulin injection. (4) that the container should be gently shaken before a dose
is withdrawn; (5) the storage conditions.
Isophane Insulin Injection

Isophane Insulin; Isophane Insulin (NPH) Protamine Zinc Insulin Injection
Isophane insulin injection is a sterile suspension of bovine, Protamine Zinc Insulin
porcine or human insulin, complexed with protamine sulphate Protamine Zinc Insulin Injection is a sterile buffered
or another suitable protamine. suspension of porcine, bovine or human insulin in the form of
614
IP 2007 INSULIN ZINC SUSPENSION
a complex obtained by the addition of Protamine Sulphate or Storage. Store in multiple dose containers in a refrigerator
another suitable protamine and Zinc Chloride or another (2° to 8°). It should not be allowed to freeze.
suitable zinc salt. Labelling. The label states (1) the strength in terms of the
Protamine Zinc Insulin Injection contains not less than 90.0 number of Units per ml; (2) the animal source or sources of the
per cent and not more than 110.0 per cent of the stated number insulin; (3) that the preparation should not be allowed to freeze;
of Units of Insulin. (4) that the container should be gently shaken before a dose
Production
It may be prepared by assaying a sterile solution of crystalline
insulin having a potency not less than 23 Units per mg, Insulin Zinc Suspension
calculated on the dried basis, adjusting its potency so that
when diluted with the other constituents in sterile form, it I.Z.S; Insulin Zinc Suspension (Mixed); I.Z.S (Mixed);
contains the requisite number of Units per ml, and adding Insulin Lente
aseptically to it a suitable protamine in the proportion of 1.0 to Insulin Zinc Suspension is a sterile, buffered suspension of
1.7 mg of protamine sulphate for each 100 Units of Insulin. It Insulin in the form of a complex obtained by the addition of
contains Sodium Phosphate as buffering agent, sufficient of a zinc chloride to insulin in a manner such that the insulin is in
suitable substance to render the preparation isotonic with a form insoluble in water. It may be prepared by mixing
blood and a sufficient amount of a suitable bactericide. It is aseptically about 3 volumes of Insulin Zinc Suspension
distributed aseptically into sterile containers which are then (Amorphous) and about 7 volumes of Insulin Zinc Suspension
sealed so as to exclude micro-organisms. (Crystalline) and distributing the mixture aseptically into sterile
Description. A white suspension which on standing deposits containers which are then sealed so as to exclude micro-
a white sediment and leaves an almost colourless supernatant organisms.
liquid. The sediment is readily resuspended on gentle shaking.
Insulin Zinc Suspension contains not less than 90.0 per cent
When examined under a microscope, approximately half the
and not more than 110.0 per cent of the stated number of Units
number of particles are seen to have no uniform shape, with a
of Insulin.
maximum diameter rarely more than 2 µm. The remaining
particles are seen to be rod-shaped crystals, the majority with Description. A white suspension which on standing deposits
a maximum diameter more than 10 µm but rarely more than a white sediment and leaves an almost colourless supernatant
100 µm. liquid. The sediment is readily resuspended on gentle shaking.
When examined under a microscope, the majority of the
Identification particles in the suspension are seen as rhombohedral crystals,
with a maximum dimension greater than 10 mm but rarely
In the Assay, the principal peak due to insulin in the exceeding 40 mm; a considerable number of particles have no
chromatogram obtained with the test solution corresponds to uniform shape and do not exceed 2 mm in maximum dimension.
the peak in the chromatogram obtained with the appropriate
reference solution. Identification
Tests In the Assay, the principal peak due to insulin in the

chromatogram obtained with test solution has a retention time
pH (2.4.24). 6.9 to 7.5. similar to that of the principal peak in the chromatogram
obtained with reference solution (a), reference (b) or reference
Total zinc. Not more than 40 µg per 100 Units of insulin,
solution (c), as appropriate.
Preparations. Tests
Other tests. Complies with the requirements stated under
pH (2.4.24). 6.9 to 7.5.
Insulin Preparations.
Total zinc. Not more than 0.0095 per cent w/v (for preparations
containing 40 Units per ml) and not more than 0.014 per cent
Test solution. To 10 ml of the preparation under examination w/v (for preparations containing 80 Units per ml), determined
add 40 µl of 5 M hydrochloric acid, mix well, allow to stand for by the following method. Take a volume of the well-shaken
1 hour to ensure solution of the sediment and dilute with suspension containing 200 Units of insulin and add 1 ml of
0.03 M hydrochloric acid to obtain a solution containing 0.1M hydrochloric acid, 10 ml of alkaline borate buffer pH
40 Units per ml. 9.0, 1 ml of 0.1M sodium hydroxide, 2 ml of a 0.0009 per cent
615
INSULIN ZINC SUSPENSION IP 2007
w/v solution of trypsin in 0.01M hydrochloric acid. Mix, allow Test solution. (for preparations containing 100 Units per ml)
to stand for 10 minutes and add 3 ml of zincon solution and centrifuge a well-shaken suspension of the preparation under
sufficient water to produce 50 ml. Allow to stand for 1 hour examination, transfer 2.0 ml of the supernatant liquid to a 5-ml
and measure the absorbance of the resulting solution at about volumetric flask, dilute to volume with 0.03 M hydrochloric
620 nm (2.4.7), using as the blank a solution prepared by acid and mix well.
treating 5 ml of water instead of the substance under Reference solution (a). Prepare as test solution but using 2.5
examination in a similar manner. Calculate the content of zinc ml of the supernatant liquid in place of 2.0 ml.
from the absorbance obtained by repeating the procedure
using a suitable aliquot of a mixture of 4 volumes of zinc Reference solution (b). Use 5 ml of the supernatant liquid.
sulphate solution and 6 volumes of water. Reference solution (c). Weigh accurately 4.5 mg of bovine
Zinc in solution. Not more than 70 per cent of the total zinc insulin RS into a 100-ml volumetric flask containing 50 ml of
(for preparations containing 40 Units per ml) and not more 0.025 M hydrochloric acid, dissolve by shaking for 5 minutes,
than 55 per cent of the total zinc (for preparations containing dilute to volume with 0.025 M hydrochloric acid and mix to
80 Units per ml), determined by the method described in the obtain a solution containing approximately 1 Unit per ml.
test for Total zinc, using 1 ml of the clear supernatant liquid Chromatographic system as described in the Assay.
obtained by centrifuging and beginning at the words “add 1 The area of the peak due to insulin in the chromatogram
ml of 0.1M hydrochloric acid,.......”. obtained with test solution, reference solution (a) or reference
Insulin extractable with buffered acetone solution. 27 per cent (b), as the case may be, is not more than that of the principal
to 40 per cent, determined by the following method. Centrifuge peak in the chromatogram obtained with reference solution
a volume containing 400 Units and reject the supernatant (c).
liquid. Suspend the residue in 3.3 ml of water, add 6.6 ml of
buffered acetone solution, stir for 3 minutes and again
centrifuge. Transfer the supernatant liquid as completely as
possible to a long-necked, round-bottomed flask, add 0.3 g of Assay. Determine by liquid chromatography (2.4.14).
nitrogen-free mercuric oxide, 3 g of anhydrous sodium Test solution. To 10 ml of the preparation under examination
sulphate, and 6 ml of nitrogen-free sulphuric acid, heat over add 40 ml of 5 M hydrochloric acid, mix well, allow to stand
a low flame until the liquid is colourless and boil for a further for 1 hour to ensure solution of the sediment and dilute with
30 minutes. Allow to cool, dilute carefully with water, add 1 g 0.03 M hydrochloric acid to obtain a solution containing 20
of zinc powder, shake and allow to stand for 10 minutes. Add Units per ml.
an excess of sodium hydroxide solution, immediately connect Reference solution (a). A 0.08 per cent w/v of bovine insulin
the flask to an ammonia distillation apparatus, mix the contents RS in 0.025 M hydrochloric acid.
and distil the liberated ammonia into 20 ml of 0.005 M sulphuric
acid prepared with carbon dioxide-free water. Rinse the Reference solution (b). A 0.08 per cent w/v of porcine insulin
condenser tube into the flask containing the distillate, add RS in 0.025 M hydrochloric acid.
sufficient carbon dioxide-free water to produce a total volume Reference solution (c). A solution containing 0.04 per cent w/
of about 50 ml and titrate the excess of sulphuric acid with v of bovine insulin RS and 0.04 per cent w/v of porcine insulin
0.01 M sodium hydroxide to pH 6.0, using a glass electrode. RS in 0.025 M hydrochloric acid for a preparation containing
Centrifuge a further volume containing 400 Units and reject both bovine and pork insulin.
the supernatant liquid. Dissolve the residue in 10 ml of a 5 per Reference solution (d). A 0.04 per cent w/v each of human
cent w/v solution of nitrogen-free sulphuric acid, transfer to insulin RS and porcine insulin RS in 0.025 M hydrochloric
a long-necked, round-bottomed flask, and repeat the acid.
procedure described above beginning at the words “add 0.3 g
of nitrogen-free mercuric oxide,.....”. Calculate the percentage
of insulin extractable with the buffered acetone solution from
the formula 100A/B, where A is the volume of 0.005 M
µm) (Ultrasphere ODS is suitable),
sulphuric acid used in the first determination and B is the
– column. temperature 45°,
volume used in the second determination.
– mobile phase. a mixture of 72.5 volumes of 0.1 M sodium
The result of the test is not valid unless in carrying out the dihydrogen phosphate adjusted to pH 2.0 with
first determination omitting the insulin preparation, not more phosphoric acid and 27.5 volumes of acetonitrile,
than 0.2 ml of 0.005 M sulphuric acid is required. – flow rate 1 ml per minute.
Insulin in solution. Determine by liquid chromatography – spectrophotometer set at 214 nm,
(2.4.14). – a 50 ì l loop injector.
616
IP 2007 INSULIN ZINC SUSPENSION (AMORPHOUS)
The test is not valid unless the resolution factor between the potency not less than 23 Units per mg, calculated on the dried
peaks corresponding to human insulin and porcine insulin is basis, a suitable quantity of zinc chloride, an appropriate
at least 1.2 in the chromatogram obtained with reference amount of a suitable substance to render the preparation
solution (d). If necessary, adjust the concentration of isotonic with blood and a sufficient quantity of a suitable
acetonitrile in the mobile phase by slight decrease or increase bactericide. It is distributed aseptically into sterile containers
until the required resolution is obtained. In the chromatogram which are then sealed so as to exclude micro-organisms.
obtained with reference solution (d) the two principal peaks, Insulin Zinc Suspension (Amorphous) contains not less than
in order of emergence, are due to human insulin and porcine 90.0 per cent and not more than 110.0 per cent of the stated
insulin and any smaller peaks appearing immediately following number of Units of Insulin.
each of the principal peaks are due to the corresponding
monodesamido derivatives. Description. A white suspension which on standing deposits
a white sediment and leaves an almost colourless supernatant
Inject either of reference solutions (a), (b) and (c), as liquid. The sediment is readily resuspended on gentle shaking.
appropriate, alternatively six times. The test is not valid unless When examined under a microscope, the particles in the
the relative standard deviation of the area of the principal suspension are seen to have no uniform shape and rarely
peak is not more than 2 per cent. exceed 2 mm in maximum dimension.
Inject the test solution. If necessary, make further adjustments
in the composition of the mobile phase so that the antimicrobial Identification
preservatives present in test solution are well separated from In the Assay, the principal peak due to insulin in the
insulin and show shorter retention times. A small reduction in chromatogram obtained with test solution has a retention time
the concentration of acetonitrile increases the retention time similar to that of the principal peak in the chromatogram
of insulin peak relatively more than those of the preservatives. obtained with reference solution (a), reference (b) or reference
If necessary, after having carried out the chromatography of a solution (c), as appropriate.
solution, wash the column with a mixture of equal volumes of
acetonitrile and water for a sufficient time in order to elute Tests
any interfering substances before injecting the next solution.
pH (2.4.24). 6.9 to 7.5.
Calculate the content of insulin from the area of the peaks due
Total zinc. Not more than 0.0095 per cent w/v (for preparations
to the bovine, porcine or human insulin and that of any peak
containing 40 Units per ml) and not more than 0.014 per cent
due to the monodesamido derivative of the insulin from the
w/v (for preparations containing 80 Units per ml), determined
declared content of insulin in bovine insulin RS, porcine
by the following method. Take a volume of the well-shaken
insulin RS or human insulin RS, as appropriate. For
suspension containing 200 Units of insulin and add 1 ml of
preparations containing both bovine and porcine insulin use
0.1 M hydrochloric acid, 10 ml of alkaline borate buffer pH
the sum of the areas of both the bovine and porcine insulin
9.0, 1 ml of 0.1 M sodium hydroxide, 2 ml of a 0.0009 per cent
peaks and of any peak due to the desamido derivative of
w/v solution of trypsin in 0.01 M hydrochloric acid. Mix,
either insulin.
allow to stand for 10 minutes and add 3 ml of zincon solution
Storage. Store in multiple dose containers at a temperature and sufficient water to produce 50 ml. Allow to stand for 1
between 2° and 8°. It should not be allowed to freeze. hour and measure the absorbance of the resulting solution at
Labelling. The label states (1) the strength in terms of the about 620 nm (2.4.7), using as the blank a solution prepared
number of Units per ml; (2) the animal source or sources of the by treating 5 ml of water instead of the substance under
insulin; (3) that the preparation should not be allowed to freeze; examination in a similar manner. Calculate the content of zinc
(4) that the container should be gently shaken before a dose from the absorbance obtained by repeating the procedure
is withdrawn; (5) the storage conditions. using a suitable aliquot of a mixture of 4 volumes of zinc
sulphate solution and 6 volumes of water.
Zinc in solution. Not more than 70 per cent of the total zinc
Insulin Zinc Suspension (Amorphous) (for preparations containing 40 Units per ml) and not more
than 55 per cent of the total zinc (for preparations containing
Amorph. I.Z.S.; Prompt Insulin Zinc Suspension 80 Units per ml), determined by the method described in the
Insulin Zinc Suspension (Amorphous) is a sterile, buffered test for Total zinc, using 1 ml of the clear supernatant liquid
suspension of Insulin in the form of a complex obtained by obtained by centrifuging and beginning at the words “add 1
the addition of zinc chloride to insulin in a manner such that ml of 0.1 M hydrochloric acid,.......”.
the solid phase of the suspension is amorphous. It may be Insulin in solution. Determine by liquid chromatography
prepared by adding aseptically to crystalline insulin having a (2.4.14).
617
INSULIN ZINC SUSPENSION (CRYSTALLINE) IP 2007
Test solution. (for preparations containing 100 Units per ml) – a 50 ì l loop injector.
centrifuge a well-shaken suspension of the preparation under The test is not valid unless the resolution factor between the
examination, transfer 2.0 ml of the supernatant liquid to a 5-ml peaks corresponding to human insulin and porcine insulin is
volumetric flask, dilute to volume with 0.03 M hydrochloric at least 1.2 in the chromatogram obtained with reference
acid and mix well. solution (d). If necessary, adjust the concentration of
Reference solution (a). Prepare as test solution but using 2.5 acetonitrile in the mobile phase by slight decrease or increase
ml of the supernatant liquid in place of 2.0 ml. until the required resolution is obtained. In the chromatogram
Reference solution (b). Use 5 ml of the supernatant liquid. obtained with reference solution (d) the two principal peaks,
in order of emergence, are due to human insulin and porcine
Reference solution (c). Weigh accurately 4.5 mg of bovine insulin and any smaller peaks appearing immediately following
insulin RS into a 100-ml volumetric flask containing 50 ml of each of the principal peaks are due to the corresponding
0.025 M hydrochloric acid, dissolve by shaking for 5 minutes, monodesamido derivatives.
dilute to volume with 0.025 M hydrochloric acid and mix to
obtain a solution containing approximately 1 Unit per ml. Inject either of reference solutions (a), (b) and (c), as
appropriate, alternatively six times. The test is not valid unless
Chromatographic system as described in the Assay. the relative standard deviation of the area of the principal
The area of the peak due to insulin in the chromatogram peak is not more than 2 per cent.
obtained with test solution, reference solution (a) or reference Inject the test solution. If necessary, make further adjustments
(b), as the case may be, is not more than that of the principal in the composition of the mobile phase so that the antimicrobial
peak in the chromatogram obtained with reference solution preservatives present in test solution are well separated from
(c). insulin and show shorter retention times. A small reduction in
Other tests. Complies with the tests stated under Parenteral the concentration of acetonitrile increases the retention time
Preparations (Injections). of insulin peak relatively more than those of the preservatives.
Assay. Determine by liquid chromatography (2.4.14). If necessary, after having carried out the chromatography of a
Test solution. To 10 ml of the preparation under examination acetonitrile and water for a sufficient time in order to elute
add 40 ml of 5 M hydrochloric acid, mix well, allow to stand any interfering substances before injecting the next solution.
for 1 hour to ensure solution of the sediment and dilute with
0.03 M hydrochloric acid to obtain a solution containing 20 Calculate the content of insulin from the area of the peaks due
Units per ml. to the bovine, porcine or human insulin and that of any peak
Reference solution (a). A 0.08 per cent w/v of bovine insulin declared content of insulin in bovine insulin RS, porcine
RS in 0.025 M hydrochloric acid. insulin RS or human insulin RS, as appropriate. For
Reference solution (b). A 0.08 per cent w/v of porcine insulin preparations containing both bovine and porcine insulin use
RS in 0.025 M hydrochloric acid. the sum of the areas of both the bovine and porcine insulin
peaks and of any peak due to the desamido derivative of
either insulin.
v of bovine insulin RS and 0.04 per cent w/v of porcine insulin
RS in 0.025 M hydrochloric acid for a preparation containing Storage. Store in multiple dose containers at a temperature
both bovine and pork insulin. between 2° and 8°. It should not be allowed to freeze.
Reference solution (d). A 0.04 per cent w/v each of human Labelling. The label states (1) the strength in terms of the
insulin RS and porcine insulin RS in 0.025 M hydrochloric number of Units per ml; (2) the animal source or sources of the
acid. insulin; (3) that the preparation should not be allowed to freeze;
Chromatographic system (4) that the container should be gently shaken before a dose
– a stainless steel column 25 cm ´ 4.6 mm packed with is withdrawn; (5) the storage conditions.
µm),
– column. temperature 45°, Insulin Zinc Suspension (Crystalline)
– mobile phase. a mixture of 72.5 volumes of 0.1 M sodium
dihydrogen phosphate adjusted to pH 2.0 with Cryst. I.Z.S.; Extended Insulin Zinc Suspension
phosphoric acid and 27.5 volumes of acetonitrile, Insulin Zinc Suspension (Crystalline) is a sterile, buffered
– flow rate 1 ml per minute. suspension of Insulin in the form of a complex obtained by
– spectrophotometer set at 214 nm, the addition of zinc chloride to insulin in a manner such that
618
IP 2007 INSULIN ZINC SUSPENSION (CRYSTALLINE)
the insulin is in the form of crystals insoluble in water. It may than 55 per cent of the total zinc (for preparations containing
be prepared by adding aseptically to crystalline insulin having 80 Units per ml), determined by the method described in the
a potency not less than 23 Units per mg, calculated with test for Total zinc, using 1 ml of the clear supernatant liquid
reference to the dried substance, a suitable quantity of zinc obtained by centrifuging and beginning at the words “add 1
chloride, an appropriate amount of a suitable substance to ml of 0.1 M hydrochloric acid,.......”.
render the preparation isotonic with blood and a sufficient Insulin extractable with buffered acetone solution. Not more
quantity of a suitable bactericide. The solution is partially than 15 per cent, determined by the following method.
neutralised to allow crystallisation to occur and the pH of the Centrifuge a volume containing 400 Units and reject the
crystalline suspension is adjusted to about 7.2. The supernatant liquid. Suspend the residue in 3.3 ml of water,
suspension is distributed aseptically into sterile containers add 6.6 ml of buffered acetone solution, stir for 3 minutes and
which are then sealed so as to exclude micro-organisms. again centrifuge. Transfer the supernatant liquid as completely
Insulin Zinc Suspension (Crystalline) contains not less than as possible to a long-necked, round-bottomed flask, add 0.3 g
90.0 per cent and not more than 110.0 per cent of the stated of nitrogen-free mercuric oxide, 3 g of anhydrous sodium
number of Units of Insulin. sulphate, and 6 ml of nitrogen-free sulphuric acid, heat over
a low flame until the liquid is colourless and boil for a further
Description. A white suspension which on standing deposits
30 minutes. Allow to cool, dilute carefully with water, add 1 g
a white sediment and leaves an almost colourless supernatant
of zinc powder, shake and allow to stand for 10 minutes. Add
liquid. The sediment is readily resuspended on gentle shaking.
an excess of sodium hydroxide solution, immediately connect
When examined under a microscope, the particles in the
the flask to an ammonia distillation apparatus, mix the contents
suspension are seen to be rhombohedral crystals, the majority
and distil the liberated ammonia into 20 ml of 0.005 M sulphuric
having a maximum dimension greater than 10 mm but rarely
acid prepared with carbon dioxide-free water. Rinse the
exceedings 40 mm.
condenser tube into the flask containing the distillate, add
sufficient carbon dioxide-free water to produce a total volume
Identification
of about 50 ml and titrate the excess of sulphuric acid with
In the Assay, the principal peak due to insulin in the 0.01 M sodium hydroxide to pH 6.0, using a glass electrode.
chromatogram obtained with test solution has a retention time Centrifuge a further volume containing 400 Units and reject
similar to that of the principal peak in the chromatogram the supernatant liquid. Dissolve the residue in 10 ml of a 5 per
obtained with reference solution (a), reference (b) or reference cent w/v solution of nitrogen-free sulphuric acid, transfer to
solution (c), as appropriate. a long-necked, round-bottomed flask, and repeat the
procedure described above beginning at the words “add 0.3 g
Tests of nitrogen-free mercuric oxide,.....”. Calculate the percentage
of insulin extractable with the buffered acetone solution from
pH (2.4.24). 6.9 to 7.5. the formula 100A/B, where A is the volume of 0.005 M
Total zinc. Not more than 0.0095 per cent w/v (for preparations sulphuric acid used in the first determination and B is the
containing 40 Units per ml) and not more than 0.014 per cent volume used in the second determination.
w/v (for preparations containing 80 Units per ml), determined The result of the test is not valid unless in carrying out the
by the following method. Take a volume of the well-shaken first determination omitting the insulin preparation, not more
suspension containing 200 Units of insulin and add 1 ml of than 0.2 ml of 0.005 M sulphuric acid is required.
0.1 M hydrochloric acid, 10 ml of alkaline borate buffer pH
9.0, 1 ml of 0.1 M sodium hydroxide, 2 ml of a 0.0009 per cent Insulin in solution. Determine by liquid chromatography
w/v solution of trypsin in 0.01 M hydrochloric acid. Mix, (2.4.14).
allow to stand for 10 minutes and add 3 ml of zincon solution Test solution. (for preparations containing 100 Units per ml)
and sufficient water to produce 50 ml. Allow to stand for 1 centrifuge a well-shaken suspension of the preparation under
hour and measure the absorbance of the resulting solution at examination, transfer 2.0 ml of the supernatant liquid to a 5-ml
about 620 nm (2.4.7), using as the blank a solution prepared volumetric flask, dilute to volume with 0.03 M hydrochloric
by treating 5 ml of water instead of the substance under acid and mix well.
examination in a similar manner. Calculate the content of zinc
from the absorbance obtained by repeating the procedure Reference solution (a). Prepare as test solution but using 2.5
using a suitable aliquot of a mixture of 4 volumes of zinc ml of the supernatant liquid in place of 2.0 ml.
sulphate solution and 6 volumes of water. Reference solution (b). Use 5 ml of the supernatant liquid.
Zinc in solution. Not more than 70 per cent of the total zinc Reference solution (c). Weigh accurately 4.5 mg of bovine
(for preparations containing 40 Units per ml) and not more insulin RS into a 100-ml volumetric flask containing 50 ml of
619
INVERT SUGAR INJECTION IP 2007
0.025 M hydrochloric acid, dissolve by shaking for 5 minutes, each of the principal peaks are due to the corresponding
dilute to volume with 0.025 M hydrochloric acid and mix to monodesamido derivatives.
obtain a solution containing approximately 1 Unit per ml. Inject either of reference solutions (a), (b) and (c), as
Chromatographic system as described in the Assay. appropriate, alternatively six times. The test is not valid unless
the relative standard deviation of the area of the principal
The area of the peak due to insulin in the chromatogram
peak is not more than 2 per cent.
obtained with test solution, reference solution (a) or reference
(b), as the case may be, is not more than that of the principal Inject the test solution. If necessary, make further adjustments
peak in the chromatogram obtained with reference solution in the composition of the mobile phase so that the antimicrobial
(c). preservatives present in test solution are well separated from
insulin and show shorter retention times. A small reduction in
the concentration of acetonitrile increases the retention time
of insulin peak relatively more than those of the preservatives.
Assay. Determine by liquid chromatography (2.4.14). If necessary, after having carried out the chromatography of a
acetonitrile and water for a sufficient time in order to elute
add 40 ml of 5 M hydrochloric acid, mix well, allow to stand
any interfering substances before injecting the next solution.
for 1 hour to ensure solution of the sediment and dilute with
0.03 M hydrochloric acid to obtain a solution containing 20 Calculate the content of insulin from the area of the peaks due
Units per ml. to the bovine, porcine or human insulin and that of any peak
Reference solution (a). A 0.08 per cent w/v of bovine insulin
declared content of insulin in bovine insulin RS, porcine
RS in 0.025 M hydrochloric acid.
insulin RS or human insulin RS, as appropriate. For
Reference solution (b). A 0.08 per cent w/v of porcine insulin preparations containing both bovine and porcine insulin use
RS in 0.025 M hydrochloric acid. the sum of the areas of both the bovine and porcine insulin
Reference solution (c). A solution containing 0.04 per cent w/ peaks and of any peak due to the desamido derivative of
v of bovine insulin RS and 0.04 per cent w/v of porcine insulin either insulin.
RS in 0.025 M hydrochloric acid for a preparation containing Storage. Store in multiple dose containers at a temperature
both bovine and pork insulin. between 2° and 8°. It should not be allowed to freeze.
Reference solution (d). A 0.04 per cent w/v each of human Labelling. The label states (1) the strength in terms of the
insulin RS and porcine insulin RS in 0.025 M hydrochloric number of Units per ml; (2) the animal source or sources of the
acid. insulin; (3) that the preparation should not be allowed to freeze;
(4) that the container should be gently shaken before a dose
µm),
– column. temperature 45o, Invert Sugar Injection
– mobile phase. a mixture of 72.5 volumes of 0.1 M sodium Invert Sugar Injection is a sterile solution of a mixture of equal
dihydrogen phosphate adjusted to pH 2.0 with amounts of Dextrose and Fructose in Water for Injections, or
phosphoric acid and 27.5 volumes of acetonitrile, an equivalent sterile solution produced by the hydrolysis of
– flow rate 1 ml per minute. Sucrose in Water for Injections. It contains no antimicrobial
– spectrophotometer set at 214 nm, agent.
– a 50 ì l loop injector.
Invert Sugar Injection contains not less than 95.0 per cent and
The test is not valid unless the resolution factor between the not more than 105.0 per cent of the labelled amount of C6H12O6.
peaks corresponding to human insulin and porcine insulin is
at least 1.2 in the chromatogram obtained with reference Description. A clear, colourless or faintly straw-coloured
solution (d). If necessary, adjust the concentration of solution.
acetonitrile in the mobile phase by slight decrease or increase
until the required resolution is obtained. In the chromatogram
Identification
obtained with reference solution (d) the two principal peaks, To 1 ml add 0.05 ml of potassium cupri-tartrate solution; the
in order of emergence, are due to human insulin and porcine solution remains blue and clear. Heat to boiling, a copious red
insulin and any smaller peaks appearing immediately following precipitate is formed.
620
IP 2007 INVERT SUGAR AND SODIUM CHLORIDE INJECTION
Tests Other tests. Complies with the tests stated under Parenteral
pH (2.4.24). 3.0 to 6.5.
Assay. Transfer 50.0 ml of cupri-tartaric solution into a 400-
5-Hydroxymethylfurfural and Related substances. Dilute a ml beaker, add 48 ml of water, mix and add 2.0 ml of the
volume containing 1.0 g of invert sugar to 500.0 ml with water preparation under examination that has been diluted
and measure the absorbance (2.4.7) of the resulting solution quantitatively with water, if necessary, to a 5.0 per cent
at the maximum at about 284 nm; absorbance at about 284 nm, concentration. Cover the beaker with a watch glass, heat the
not more than 0.25. solution, regulating the heat so that boiling begins in 4 minutes
Heavy metals (2.3.13). A solution prepared by evaporating a and continue boiling for 2 minutes. Filter the hot solution at
volume containing 4.0 g of invert sugar to 10 ml and adding 2 once through a tared porcelain filtering crucible, wash the
ml of dilute acetic acid and sufficient water to produce 25 ml precipitate with water maintained at 60°, then with 10 ml of
complies with the limit test for heavy metals, Method A (5 ethanol (95 per cent). Dry at 105° to constant weight. Carry
ppm). out a blank determination and make any necessary correction.
The corrected weight of the precipitate so obtained is not less
Chlorides (2.3.12). 2.0 ml of the injection complies with the
than 204.0 mg and not more than 224.4 mg.
limit test for chlorides (120 ppm).
exceeding 30°.
per ml.
Labelling. The label states (1) whether it is produced by
Completeness of inversion. hydrolysis of Sucrose or by mixing Dextrose and Fructose; (2)
NOTE — Invert Sugar Injection that is produced by mixing the strength as the percentage w/v of invert sugar; (3) total
Dextrose and Fructose is exempt from this test. osmolar concentration in mOsmol per litre; (4) that the injection
should not be used if it contains visible particles.
Test solution. Transfer an accurately measured volume of the
preparation under examination containing about 2.5 g of invert
sugar to a 100-ml volumetric flask, dilute to volume with water
Invert Sugar and Sodium Chloride
and mix. Injection
Reference solution. Prepare a solution in water containing Sodium Chloride and Invert Sugar Intravenous Infusion
known concentrations of about 0.25 mg of sucrose and about
Invert Sugar and Sodium Chloride Injection is a sterile solution
12.5 mg of dextrose per ml.
of a mixture of equal amounts of Dextrose and Fructose in
Chromatographic system Water for Injections to which the required amount of Sodium
– a stainless steel column 30 cm x 7.8 mm, packed with a Chloride is added. Invert sugar may be prepared by acid
strong cation-exchange resin consisting of sulphonated hydrolysis of Sucrose.
Invert Sugar and Sodium Chloride Injection contains not less
calcium form (9 µm),
than 95.0 per cent and not more than 105.0 per cent of the
– mobile phase: filtered and degassed water,
stated amounts of sodium chloride, NaCl, and invert sugar,
– column temperature. constant at about 40°,
– refractive index detector, Description. A clear, colourless or faintly straw-coloured
– a 20 µl loop injector. solution.
Inject the reference solution. The sucrose elutes first and the Identification
peak is baseline separated from the dextrose peak. The relative
standard deviation for replicate injections is not more than A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
2.0 per cent. the solution remains blue and clear. Heat to boiling, a copious
red precipitate is formed.
Inject the test solution and the reference solution and measure
the responses for the sucrose peak. Calculate the content of B. Gives reaction A of chlorides and reaction B of sodium salts
sucrose in the volume taken of the preparation under (2.3.1).
examination. Not more than 1.5 per cent of the quantity of
Tests
invert sugar in the volume taken of the preparation under
examination, based on the value stated on the label, is found. pH (2.4.24). 3.0 to 6.5.
621
IODINE IP 2007
5-Hydroxymethylfurfural and Related substances. Dilute a 1 ml of 0.1 M silver nitrate is equivalent to 0.005844 g of NaCl.
volume containing 1.0 g of invert sugar to 500.0 ml with water For invert sugar — Transfer 50.0 ml of cupri-tartaric solution
and measure the absorbance (2.4.7) of the resulting solution into a 400-ml beaker, add 48 ml of water, mix and add 2.0 ml of
at the maximum at about 284 nm; absorbance at about 284 nm, the preparation under examination that has been diluted
not more than 0.25. quantitatively with water, if necessary, to a 5.0 per cent
Completeness of inversion concentration. Cover the beaker with a watch glass, heat the
solution, regulating the heat so that boiling begins in 4 minutes
NOTE — Invert Sugar and Sodium Chloride Injection that is
and continue boiling for 2 minutes. Filter the hot solution at
produced by mixing Dextrose and Fructose is exempt from
once through a tared porcelain filtering crucible, wash the
this test.
precipitate with water maintained at 60°, then with 10 ml of
Determine by liquid chromatography (2.4.14). ethanol (95 per cent). Dry at 105° to constant weight. Carry
Test solution. Transfer an accurately measured volume of the out a blank determination and make any necessary correction.
preparation under examination containing about 2.5 g of invert The corrected weight of the precipitate so obtained is not less
sugar to a 100-ml volumetric flask, dilute to volume with water than 204.0 mg and not more than 224.4 mg.
and mix. Storage. Store in single dose containers at a temperature not
Reference solution. Prepare a solution in water containing exceeding 30°.
known concentrations of about 0.25 mg of sucrose and about Labelling. The label states (1) whether it is produced by
12.5 mg of dextrose per ml. hydrolysis of Sucrose or by mixing Dextrose and Fructose; (2)
Chromatographic system the strength as the percentage w/v of sodium chloride and
– a stainless steel column 30 cm x 7.8 mm, packed with a invert sugar; (3) total osmolar concentration in mOsmol per
strong cation-exchange resin consisting of sulphonated litre; (4) approximate concentrations, in millimoles per litre, of
cross-linked styrene-divinylbenzene copolymer in the the sodium and chloride ions; (5) that the injection should not
calcium form (9 µm), be used if it contains visible particles.
– mobile phase: filtered and degassed water,
– column temperature. constant at about 40°,
– refractive index detector,
Iodine
– a 20 µl loop injector. I2 Mol. Wt. 253.8
Inject the reference solution. The sucrose elutes first and the Iodine contains not less than 99.5 per cent and not more
peak is baseline separated from the dextrose peak. The relative than 100.5 per cent of I.
standard deviation for replicate injections is not more than 2.0
per cent. Description. Greyish violet brittle plates or small crystals with
a metallic sheen; odour, irritant. It volatalises slowly at room
Inject the test solution and the reference solution and measure temperature.
the responses for the sucrose peak. Calculate the content of
sucrose in the volume taken of the preparation under Identification
examination. Not more than 1.5 per cent of the quantity of
A. When heated gently it gives violet vapours which condense
invert sugar in the volume taken of the preparation under
forming a bluish-black crystalline sublimate.
examination, based on the value stated on the label, is found.
Heavy metals (2.3.13). A solution prepared by evaporating a B. A saturated solution yields a blue colour in the presence of
volume containing 4.0 g of invert sugar to 10 ml and adding 2 starch solution which disappears when the solution is heated
ml of dilute acetic acid and sufficient water to produce 25 ml and reappears when it is cooled.
complies with the limit test for heavy metals, Method A (5 Tests
ppm).
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit Bromides and chlorides. Not more than 250 ppm, determined
per ml. by the following method. Triturate 3.0 g with 20 ml of water,
filter, wash the filter, dilute the filtrate to 30 ml with water and
Other tests. Complies with the tests stated under Parenteral add 1 g of zinc powder. When the solution is decolorised,
Preparations (Injections). filter and wash the filter with sufficient water to produce 40 ml
Assay. For sodium chloride — Titrate an accurately measured of filtrate. To 10 ml of this solution add 3 ml of 10 M ammonia
volume containing about 0.1 g of Sodium Chloride with 0.1 M and 6 ml of silver nitrate solution, filter, wash the filter with
silver nitrate using potassium chromate solution as indicator. water and dilute to 20 ml with water. To 10 ml of the filtrate add
622
IP 2007 IRINOTECAN HYDROCHLORIDE TRIHYDRATE
1.5 ml of nitric acid. After 1 minute any opalescence produced Related substances. Determine by liquid chromatography
is not more intense than that produced in a solution prepared (2.4.14).
at the same time by mixing 10.75 ml of water, 0.25 ml of 0.01 M Test solution. Dissolve 100 mg of the substance under
hydrochloric acid, 0.2 ml of 2 M nitric acid and 0.3 ml of examination in 10 ml of the mobile phase.
silver nitrate solution.
Non-volatile matter. Not more than 0.1 per cent, determined
ipratropium bromide RS in the mobile phase.
by heating 1.0 g in a porcelain dish on a water-bath until the
iodine has volatilised and drying the residue at 105°. Chromatographic system
Assay. Weigh accurately about 0.2 g, transfer to a flask – a stainless steel column 15 cm x 3.9 mm, packed with
containing 1 g of potassium iodide and 2 ml of water, add 1 ml octadecylsilane bonded to porous silica (4 µm),
of 2 M acetic acid, dissolve completely and add 50 ml of – mobile phase: a mixture of 87 volumes of buffer solution
water. Titrate with 0.1 M sodium thiosulphate using starch pH 5.5 prepared by dissolving 1.24 g of sodium
solution as indicator. dihydrogen phosphate and 0.17 g of tetrapropyl
ammonium chloride in 87 ml of water and adjusting pH
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.01269 g to 5.5 with 18 per cent w/v solution of disodium
of I. hydrogen phosphate, and 13 volumes of methanol,
Storage. Store in ground-glass-stoppered containers or in – flow rate. 1.5 ml per minute,
earthenware containers with waxed bungs. – spectrophotometer set at 220 nm,
Ipratropium Bromide tailing factor is not more than 2.5.
Inject the test solution. Any individual impurity is not more
H3C than 0.5 per cent and the sum of all the impurities is not more
CH3 than 1.0 per cent.
H3C
N
Sulphated Ash (2.3.18). Not more than 0.1 per cent.
OH , Br, H2O Assay. Weigh accurately about 0.35 g of the substance under
examination, dissolve in 50 ml of water and add 3 ml of dilute
O nitric acid. Titrate with 0.1 M silver nitrate, determining the
end-point potentiometrically (2.4.25). Carry out a blank titration.
O
C20H30BrNO3.
C20H30BrNO3,H2O Mol. Wt. 430.4
Ipratropium Bromide is (1R, 3r, 5S, 8r)-3-[[(2RS)-3-hydroxy-
2-phenylpropanoyl]oxy]-8-methyl-8-(1-methylethyl)-8-
azoniabicyclo[3.2.1]octane bromide monohydrate
Ipratropium Bromide contains not less than 99.0 per cent and Irinotecan Hydrochloride Trihydrate
not more than 100.5 per cent of C20H30BrNO3,H2O, calculated
Description. A white or almost white crystalline powder. N CH3
N O O
Identification
N
O
Determine by infrared absorption spectrophotometry (2.4.6). N , HCl, 3H2 O
Compare the spectrum with that obtained with ipratropium H3C O
bromide RS or with the reference spectrum of ipratropium
bromide. HO O
Tests C33H38N4O6, HCl, 3H2O Mol. Wt. 677.18

pH (2.4.24). 5.0 to 7.5, determined in a 1.0 per cent w/v solution Irinotecan Hydrochloride Trihydrate is (4S)-4,11-diethyl-
in carbon dioxide-free water. 3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo-1H-pyrano
623
IRINOTECAN HYDROCHLORIDE TRIHYDRATE IP 2007
[3′,4′:6,7]indolizino[1,2-b]quinolin-9-yl [1,4′-bipiperidine]-1′- – mobile phase: A. dissolve 8.90 g of disodium hydrogen

carboxylate hydrochloride. phosphate dihydrate and 1.01 g of 1- heptane sulphonic
Irinotecan Hydrochloride Trihydrate contains not less than acid sodium in 900 ml of water. Adjust the pH to 3.0
98.0 per cent and not more than 102.0 per cent of with orthophosphoric acid and dilute to 1000 ml with
C33H38N4O6.HCl, calculated on the anhydrous basis. water and filter,
B. acetonitrile,
Description. A pale yellow to yellow crystalline powder. – flow rate. 1 ml per minute,
CAUTION — Irinotecan Hydrochloride Trihydrate is – a linear gradient programme using the conditions given
potentially cytotoxic. Great care should be taken in handling below,
the powder and preparing solutions. – spectrophotometer set at 254 nm,
Identification Time mobile phase A Mobile phase B
A. Determine by infrared absorption spectrophotometry (2.4.6). 0 78 22
Compare the spectrum with that obtained with irinotecan
5 78 22
hydrochloride trihydrate RS.
28 67 33
B. In the Assay, the principal peak in the chromatogram 40 67 33
45 78 22
46 78 22
Tests Reference solution (c)-( i) weigh accurately about 10 mg of 7-
ethyl-10-hydroxycamptothecin into a 25-ml volumetric flask.
Appearance of solution. A 1.0 per cent w/v solution is clear Dissolve to dilute to volume with N,N,dimethylformamide
(2.4.1) and not more intensely coloured than the reference (solution-A).weight 20 mg of irinotecon hydrochloride RS in
solution GYS3 (2.4.1). 10 ml of solution A and dilute to volume 50 ml with solvent
pH (2.4.24). 3.0 to 5.0, determined in 1.0 per cent w/v solution mixure.
in water. The system is not valid unless the solution between 7-ethyl-
Specific optical rotation (2.4.22). + 60.0º to + 73.0º, determined 10-hydroxycamptothecin.
in 1.0 per cent w/v solution in water. Inject reference solution (a). The test is not valid unless the
Light absorption. Dissolve 0.1 g of the substance under column efficiency is not less than 2000 theoretical plates and
examination in 10 ml of methanol. The absorbance of the the tailing factor is not more than 2.0.
resulting solution, at about 430 nm (2.4.7) is not more than Inject the test solution and reference solution (b). In the
0.17. chromatogram obtained with the test solution, the area of any
(2.4.14).
Solvent mixture. Dissolve 6.41 g of disodium hydrogen is not more than the area of the peak in the chromatogram
phosphate dihydrate and 0.73 g 1-heptane sulphonic acid obtained with the reference solution (b) (1.0 per cent).
sodium in 720 ml of water. Adjust the pH to 3.0 with Test solution. Weigh accurately about 0.5 g of the substance
orthophosphoric acid, dilute to 1000 ml with acetonitrile. under examination in 5 ml of N,N’-dimethyl formamide. Mix 1
Test solution. Dissolve 40 mg of the substance under ml of the solution with 5 ml of water in to 10 ml of headspace
examination in 100 ml of solvent mixture. vail.
Reference solution (a). A 0.04 per cent w/v solution of Reference solution (a). To 50 ml of water, add 12.6 µl of
irinotecan hydrochloride trihydrate RS in solvent mixture. ethanol, 126 µl of acetone, dilute to 100 ml with water.
Reference solution (b). Dilute 1 ml of reference solution (a) to Reference solution (b). Dilute 80 µl of chloroform to 10 ml
100 ml with solvent mixture. with N,N’-dimethyl formamide. Dilute 1 ml of the solution to
100 ml with water. Further, dilute 1 ml of this solution to 10 ml
NOTE — Use freshly prepared solutions. with water.
Chromatographic system Reference solution (c). Dilute 10 ml each of reference solutions
– a stainless steel column 25 cm x 4.6 mm packed with (a) and (b) to 100 ml with water. To 5 ml of this solution, add 1
octadecylsilane bonded to silica (5 µm), ml of N,N’-dimethyl formamide in to 10 ml of headspace vail.
624
IP 2007 IRINOTECAN INJECTION
Chromatographic system Assay. Determine by liquid chromatography (2.4.14).

– a capillary column 30 m x 0.53 mm, 3 µmpacked with Solvent mixture. Dissolve 6.41 g of disodium hydrogen
mega bore coated with a mixture of 6 per cent phosphate dihydrate and 0.73 g 1-heptane sulphonic acid
cyanopropylphenyl and 94 per cent dimethylpoly- sodium in 720 ml of water. Adjust the pH to 3.0 with ortho-
siloxane (3 µm ), phosphoric acid, dilute to 1000 ml with acetonitrile, filter.
– temperature:
column. 50º for 10 minutes increase @ 30º per minute to Test solution. Dissolve 40 mg of the substance under
220º hold for 15 minutes, examination in 100 ml of solvent mixture.
inlet port 180ºand detector. 260º, Reference solution. A 0.04 per cent w/v solution of irinotecan
– nitrogen as carrier gas with a flow rate. 4.8 ml per min. hydrochloride trihydrate RS in solvent mixture.
Head space conditions
– Vial equilibrium temperature 85º, loop temperature 110º , Chromatographic system
Transfer line 160º, vial equilibrium 45 minutes, vial – a stainless steel column 25 cm x 4.6 mm, packed with
pressurisation 0.5 minute, sample loop fill 0.2 minute, octadecylsilane bonded to porous silica (5 µm),
loop equilibrium 0.05 minute, sample injection 1 minute, – mobile phase: A. dissolve 8.90 g of disodium hydrogen
vial pressure 10 psi. phosphate dihydrate and 1.01 g of 1-heptane sulphonic
– a flame ionisation detector, acid sodium in 900 ml of water. Adjust the pH to 3.0
– nitrogen as carrier gas. with orthophosphoric acid and dilute to 1000 ml with
water,
Inject 1 ml of the reference solution (c). The test is not valid B. acetonitrile,
unless the resolution between the peaks due to ethanol and – flow rate.1 ml per minute,
acetone, the peaks due to acetone and chloroform is not more – a linear gradient programme using the conditions given
than 1.5 and the tailing factor is not more than 1.5 for each below,
component. – spectrophotometer set at 254 nm,
Inject 1 ml of the test solution and reference solution (c). In – a 10 µl loop injector.
the chromatogram obtained with test solution, the area of Time Mobile phase A Mobile phase B
peaks due to ethanol, acetone and chloroform is not more ( in min) (per cent v/v) (per cent v/v)
than the area of peaks obtained in 0 78 22
the chromatogram due to reference solution (c). 5 78 22
Heavy metals (2.3.13). 1.0 g complies with the limit test for 28 67 33
heavy metals, Method A (20 ppm). 40 67 33
Total Chloride. 5.0 per cent to 5.5 per cent, Weigh accurately 45 78 22
about 500 mg of the Irinotecan Hydochloride Trihydrate, 46 78 22
dissolve in 10 ml of methanol, add 20 ml of water and 20 ml of Inject the reference solution. The test is not valid unless the
glacial acetic acid and titrate with 0.1 M silver nitrate tailing factor is not more than 1.5. The relative standard
solution, using eosin yellow solution as indicator. Carry out a deviation for replicate injections is not more than 2.0 per cent.
blank titration.
Inject the test solution and reference solution.
chloride. Calculate the content of C33H38N4O6.HCl.
Storage. Store protected from light, at a temperature not
Loss on ignition (2.4.20). Not more than 0.1 per cent.
exceeding 25º.
Water (2.3.43). 8.0 per cent to 9.0 per cent, determined on
0.1 g.
Microbial Contamination (2.2.9). The total aerobic microbial Irinotecan Injection
count does not exceed 100 cfu per g. It meets the requirements
of the tests for the absence of Staphylococcus aureus, Irinotecan Hydrochloride Injection
Pseudomonas aeruginosa, Salmonella species, and Irinotecan Injection is a sterile solution of Irinotecan
Escherichia coli. Hydrochloride Trihydrate in water.
Bacterial endotoxins (2.2.3). Not more than 0.275 Endotoxin Irinotecan Injection contains not less than 90.0 per cent and
Unit per mg of irinotecan hydrochloride trihydrate. not more than 110.0 per cent of the stated amount of irinotecan
625
IRON AND AMMONIUM CITRATE IP 2007
hydrochloride, C33H38N4O6.HCl, calculated on the anhydrous Bacterial endotoxins (2.2.3). Not more than 0.556 Endotoxin
basis. Unit per mg of irinotecan hydrochloride trihydrate.
Description. A light yellow coloured solution, free from visible Sterility(2.2.11).Complies with the test for sterility.
particles.
Identification Preparations (Injections).
with the test solution corresponds to the peak in the Test solution. Accurately measured volume of the injection
chromatogram obtained with the reference solution. containing 40 mg of irinotecan hydrochloride trihydrate,
diluted to 100.0 ml with mobile phase.
A. It gives the reaction of chlorides (2.3.1).
Reference solution. A 0.04 per cent w/v solution of irinotecan
Tests hydrochloride trihydrate RS with mobile phase.
Appearance of solution( 2.4.1). Prepare a solution of equivalent Chromatographic system as described under Related
to1.0 per cent of the substance in water. The solution is clear substances.
or not more intensely coloured than the reference Inject the reference solution. The test is not valid unless the
solutionGYS3. tailing factor is not more than 1.4. The relative standard
pH (2.4.24). 3.0 to 3.8. deviation for replicate injections is not more than 2.0 per cent.
Related substances. Determine by liquid chromatography Inject the test solution and the reference solution.
(2.4.14). Calculate the content of C33H38N4O6.HCl.
Test solution. Dilute a measured volume containing 40 mg of Storage. Store protected from light, at a temperature not
Irinotecan Hydrochloride to 100 ml with mobile phase. exceeding 25º.
irinotecan hydrochloride trihydrate RS with mobile phase.
Reference solution (b). Dilute 1 ml of reference solution (a) to Iron and Ammonium Citrate
Ferric Ammonium Citrate
– a stainless steel column 25 cm x 4.6 mm, packed with Iron and Ammonium Citrate is a complex ammonium ferric
octadecylsilane bonded to porous silica (5 µm), citrate.
– mobile phase: a mixture of 72 volumes of buffer solution Iron and Ammonium Citrate contains not less than 20.5 per
prepared by dissolving 1.1 g of 1-heptane sulphonic cent and not more than 22.5 per cent of iron, Fe.
acid sodium monohydrate and 7.1 g of disodium
Description. Thin, transparent, dark red scales or granules or
hydrogen orthophosphate anhydrous in 1000 ml water.
a brownish red powder; odourlTess; deliquescent in moist air
Adjusted the pH to 3.0 with orthophosphoric acid and
and is affected by light.
28 volumes of acetonitrile, filter.
– flow rate.1.5 ml per minute, Identification
– a 10 µl loop injector. A. Ignite 0.1 g gently and dissolve the residue in 5 ml of
hydrochloric acid; the solution gives the reactions of ferric
salts (2.3.1).
the tailing factor is not more than 2.0. B. Warm 50 mg with 5 M sodium hydroxide; ammonia is
evolved and the solution gives the reactions of citrates (2.3.1).
chromatogram obtained with the test solution, the area of any Tests
in the chromatogram obtained with the reference solution (b) Arsenic (2.3.10). Mix 2.5 g with 1.5 g of anhydrous sodium
(0.2 per cent) and the sum of areas of all the secondary peaks carbonate, add 10 ml of bromine solution and mix thoroughly.
is not more than 1.2 times the area of the peak in the Evaporate to dryness on a water-bath, gently ignite and
chromatogram obtained with the reference solution (b) (1.2 dissolve the cooled residue in 20 ml of brominated
per cent). hydrochloric acid and 10 ml of water. Transfer to a small
626
IP 2007 IRON DEXTRAN INJECTION
flask, add sufficient stannous chloride AsT to remove the Assay. Weigh accurately about 0.5 g, dissolve in a mixture of
yellow colour, connect to a condenser and distil 22 ml. To the 15 ml of water and 1 ml of sulphuric acid and warm until the
distillate add 40 ml of water and 0.15 ml of stannous chloride dark brown colour becomes yellow. After cooling the solution
AsT. The resulting solution complies with the limit test for to 15° add dropwise 0.02 M potassium permanganate till a
arsenic (4 ppm). pink colour persisting for 5 seconds is obtained. Add 15 ml of
Lead. Dissolve 2.0 g in 20 ml of hydrochloric acid and 8 ml of hydrochloric acid and 2 g of potassium iodide, allow to stand
water, add 0.5 ml of nitric acid, heat just to boiling, cool, for 3 minutes, add about 60 ml of water and titrate with 0.1 M
transfer to a separating funnel and extract with three quantities, sodium thiosulphate using starch solution as indicator.
each of 20 ml, of ether. If the acid solution is still more than 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.005585 g
faintly yellow, extract with an additional 20 ml of ether. Reject of Fe.
the ether extracts. Transfer the acid solution to a narrow-
necked flask, rinse the separating funnel with 5 ml of water
and add the rinsings to the flask. Heat to remove the dissolved
ether, cool, make alkaline with ammonia solution, add 1 ml of
potassium cyanide solution, dilute to 50 ml with water and
add 0.1 ml of sodium sulphide solution. Any colour produced Iron Dextran Injection
is not more intense than that produced by mixing 10 ml of Iron Dextran Injection is a sterile colloidal solution containing
hydrochloric acid, 0.5 ml of nitric acid and 6 ml of lead a complex of ferric hydroxide with dextrans of average
standard solution (10 ppm Pb), making alkaline with ammonia molecular weight between 5000 and 7500.
solution and adding 1 ml of potassium cyanide solution and
0.1 ml of sodium sulphide solution (30 ppm). Iron Dextran Injection contains not less than 4.75 per cent and
not more than 5.25 per cent w/v of iron, Fe, and not less than
Zinc. Dissolve 2.0 g in a mixture of 20 ml of hydrochloric acid 17.0 per cent and not more than 23.0 per cent w/v of dextrans.
and 8 ml of water, add 0.5 ml of nitric acid, heat just to boiling,
cool and extract with three quantities, each of 20 ml, of ether. Description. A dark brown solution.
If the acid solution is still more than faintly yellow, repeat the
extraction with an additional 20 ml of ether. Reject the ether Identification
extracts. Warm the acid solution on a water-bath to remove
A. To 0.2 ml, previously diluted to 5 ml with water, add 5 M
dissolved ether, cool and add sufficient water to produce 100
ammonia; no precipitate is produced.
ml. To 10 ml add 1 g of citric acid and 0.1 g of resorcinol,
neutralise with dilute ammonia solution, using thymol blue B. Mix 1 ml with 100 ml of water. To 5 ml of this solution add 0.1
solution as indicator, and shake for 1 minute with two ml of hydrochloric acid, boil for 30 seconds, cool rapidly, add
successive quantities, each of 20 ml, of dithizone solution. To 2 ml of strong ammonia solution and 5 ml of hydrogen sulphide
the combined extracts add 10 ml of 0.1 M hydrochloric acid, solution, boil to remove hydrogen sulphide, cool and filter.
shake for 1 minute, separate the acid layer and wash it with 2 Boil 5 ml of the filtrate with 5 ml of potassium cupri-tartrate
ml of chloroform. To the acid layer add 3 ml of 1 M hydrochloric solution; the solution remains greenish in colour and no
acid and 20 ml of ammonium chloride solution and adjust the precipitate is produced. Boil a further 5 ml of the filtrate with
volume to 50 ml with water. Add 1 ml of potassium ferrocyanide 0.5 ml of hydrochloric acid for 5 minutes, cool, add 2.5 ml of 5
solution and allow to stand for 15 minutes. Any turbidity M sodium hydroxide and 5 ml of potassium cupri-tartrate
produced is not more than that produced by the addition of 1 solution and boil again; a reddish precipitate is produced.
ml of potassium ferrocyanide solution to a freshly prepared C. To 1 ml add 20 ml of water and 5 ml of hydrochloric acid
mixture of 1.0 ml of zinc standard solution (10 ppm Zn), 4 ml and boil for 5 minutes. Cool, add an excess of strong ammonia
of 1 M hydrochloric acid, 20 ml of ammonium chloride solution and filter. Wash the precipitate with water, dissolve
solution and sufficient water to produce 50 ml (50 ppm). in the minimum volume of 2 M hydrochloric acid and add
Free ferric compounds. A 1.0 per cent w/v solution gives no sufficient water to produce 20 ml. The resulting solution gives
blue precipitate with potassium ferrocyanide solution unless reaction B of ferric salts (2.3.1).
acidified with hydrochloric acid.
Tests
Chlorides (2.3.12). Dissolve 0.1 g in 5 ml of water and boil
with 2 ml of nitric acid; the solution complies with the limit pH (2.4.24). 5.2 to 6.5.
test for chlorides (0.25 per cent). Arsenic (2.3.10). To 10.0 ml in a round-bottomed, long-necked
Sulphates (2.3.17). 10 ml of a 0.5 per cent w/v solution complies flask add 20 ml of water and 20 ml of nitric acid and heat until
with the limit test for sulphates (0.3 per cent). the vigorous evolution of brown fumes ceases. Cool, add 20
627
IRON DEXTRAN INJECTION IP 2007
ml of sulphuric acid and heat again until fumes are evolved, and 2 g of ammonium chloride diluted to 50 ml with water and
adding nitric acid dropwise from time to time until oxidation allowed to stand for 20 minutes (150 ppm).
is complete. Cool, add 60 ml of water, bring to boil and continue
Chlorides. To 5.0 ml add 75 ml of water and 0.05 ml of nitric
boiling until the volume of liquid is reduced to about 40 ml.
acid and titrate immediately with 0.1 M silver nitrate,
Cool and dilute to 100 ml with water. Reserve a portion of the
determining the end-point potentiometrically (2.4.25). 6.8 to
solution (solution A) for the test for Heavy metals. Boil gently
9.6 ml of 0.1 M silver nitrate is required.
40 ml of this solution until the volume is reduced to about 15
ml, cool and add 15 ml of stannous chloride solution AsT. Iron absorption. Prepare a site over the semitendinosus muscle
Connect to a condenser and distil 15 ml into 25 ml of water. To of one leg of each of two rabbits, each weighing between 1.5
the distillate add 0.2 ml of bromine solution, remove the excess and 2.5 kg, by clipping the fur and disinfecting the exposed
of bromine with a few drops of stannous chloride solution skin. Inject each site with a dose of 0.4 ml per kg of body
AsT and add 20 ml of water. The resulting solution complies weight in the following manner. Place the needle in the distal
with the limit test for arsenic. Use 0.8 ml of arsenic standard end of the semitendinosus muscle at an angle such as to
solution (10 ppm As) to prepare the standard (2 ppm). ensure that the full length of the needle is used, then pass it
through the sartorius and vastus medialis muscles. House the
Heavy metals (2.3.13). To 16.0 ml of solution A reserved in the rabbits separately. Sacrifice the rabbits after 7 days and remove
test for Arsenic add 50 ml of hydrochloric acid and extract the legs into which the injection were made. Carefully dissect
with four quantities, each of 20 ml, of isobutyl acetate, the muscles and examine the site of injection. The muscle is
discarding the extracts. Evaporate the acid solution to dryness only lightly stained and no heavy black deposit of unabsorbed
and dissolve the residue in 20 ml of water. The resulting iron compounds or leakage along fascial planes is observed.
solution complies with the limit test for heavy metals, Method
D (25 ppm). Use lead standard solution (2 ppm Pb) to prepare Skin the leg, dissect the flesh from the bone and cut into small
the standard. pieces. Transfer the pieces to a 1000-ml beaker, add 75 ml of 2
M sodium hydroxide and sufficient water to submerge them,
Copper. To 5.0 ml add 5 ml of nitric acid and heat until the cover the beaker with a watch glass and boil until most of the
vigorous evolution of brown fumes ceases. Cool, add 2 ml of solid matter has disintegrated. Cool cautiously, add 50 ml of
sulphuric acid and heat again until fumes are evolved, adding sulphuric acid, heat the mixture almost to boiling and add
nitric acid dropwise at intervals until oxidation is complete. carefully 10 ml of fuming nitric acid, about 1 ml at a time, until
Cool, add 25 ml of hydrochloric acid, warm to dissolve, cool no charring occurs when the excess of nitric acid has been
and extract with four quantities, each of 25 ml, of isobutyl boiled off. Cool, add 175 ml of water, boil until solution is
acetate, discarding the extracts. Evaporate the acid solution complete, cool and dilute to 250.0 ml with water. To 5.0 ml of
to dryness, adding nitric acid dropwise if charring occurs. the solution add 3 ml of sulphuric acid, heat to fuming and
Dissolve the residue in 10 ml of 1 M hydrochloric acid, complete the oxidation by adding small quantities of nitric
reserving a portion (solution B) for the test for Zinc. To 1 ml acid until the solution is colourless. Cool, add 20 ml of water,
add 25 ml of water and 1 g of citric acid, make alkaline to boil for 3 minutes and add 10 ml of ammonium citrate solution,
litmus paper with 5 M ammonia, dilute to 50 ml with water, 10 ml of ammonium thioglycollate solution followed by dilute
add 1 ml of sodium diethyldithiocarbamate solution and allow ammonia solution dropwise until the iron colour is fully
to stand for 5 minutes. Any colour produced is not more developed. Add 1 ml excess of dilute ammonia solution and
intense than that produced by treating in the same manner a sufficient water to produce 100.0 ml. Measure the absorbance
mixture of 3 ml of copper standard solution (10 ppm Cu) and of the resulting solution at about 530 nm (2.4.7). For the
1 ml of 1 M hydrochloric acid beginning at the words “add 25 reference solution, add 10 ml of ammonium citrate solution,
ml of water.....” (60 ppm). 10 ml of ammonium thioglycollate solution and the same
Zinc. To 5.0 ml of solution B reserved in the test for Copper quantities of dilute ammonia solution as used above to 20 ml
add 15 ml of 1 M sodium hydroxide, boil, filter, wash the residue of water, and dilute to 100.0 ml. Measure the absorbance of
with water and dilute the combined filtrate and washings to this solution at about 530 nm (2.4.7). From the difference
25 ml with water. To 5 ml add 5 ml of 1 M hydrochloric acid between the absorbances, calculate the amount of Fe present
and 2 g of ammonium chloride, dilute to 50 ml with water, add in the legs from a reference curve prepared by treating suitable
1 ml of freshly prepared dilute potassium ferrocyanide solution aliquots of a solution of ferric ammonium sulphate containing
and allow to stand for 20 minutes. Any opalescence produced 0.01 per cent w/v of Fe by the above procedure beginning at
is not more than that produced when 1 ml of freshly prepared the words “add 10 ml of ammonium citrate solution,.....”.
dilute potassium ferrocyanide solution is added to a solution Repeat the determination of Fe on the corresponding legs
prepared from 3 ml of zinc standard solution (25 ppm Zn), 3 into which injection was not made beginning at the words
ml of 1 M sodium hydroxide, 6 ml of 1 M hydrochloric acid “Carefully dissect the muscles....”. From the difference between
628
IP 2007 ISONIAZID
the two amounts of Fe, calculate the proportion of injected Isoniazid

iron, as Fe, remaining in the leg tissues. Not more than 20 per
cent of the injected iron remains. Isonicotinylhydrazine; INH
Abnormal toxicity. Inject 0.10 ml into a tail vein of each of 10
mice; not more than 3 mice die within 5 days of injection. If O
more than 3 mice die within 5 days, repeat the test on another NH2
group of 20 mice. Not more than 10 of the 30 mice used in the N
N H
combined tests die within 5 days of injection.
C6H7N3O Mol. Wt. 137.1
Unit per mg of iron.
Isoniazid is isonicotinic acid hydrazide.
Preparations (Injections). Isoniazid contains not less than 98.0 per cent and not more
Assay. For iron — Weigh accurately about 2.0 g, add 10 ml of than 101.0 per cent of C6H7N3O, calculated on the dried basis.
water and 5 ml of sulphuric acid and stir for several minutes. Description. Colourless crystals or a white, crystalline powder;
Allow to stand for 5 minutes, cool and dilute to 50 ml with odourless.
water. Prepare a suitable zinc amalgam by covering 300 g of
zinc shots with a 2 per cent w/v solution of mercuric chloride Identification
and stir for 10 minutes. Decant the solution, wash the residue
three times with water and transfer it to a column (30 cm x 18 Test A may be omitted if tests B and C are carried out. Test B
mm) fitted with a sintered-glass disc (porosity No. 0). Activate may be omitted if tests A and C are carried out.
the zinc amalgam by passing through the column 200 ml of A. Determine by infrared absorption spectrophotometry (2.4.6).
sulphuric acid (5 per cent v/v). Pass the prepared solution Compare the spectrum with that obtained with isoniazid RS
slowly through the column and wash successively with 50 ml or with the reference spectrum of isoniazid.
of water, four quantities, each of 25 ml, of sulphuric acid
B. Dissolve 0.1 g in 2 ml of water, add a warm solution of 0.1 g
(5 per cent v/v) and 50 ml of water. Titrate the combined
of vanillin in 10 ml of water, allow to stand and scratch the
eluates with 0.1 M ceric ammonium sulphate using ferroin
inside of the container with a glass rod; a yellow precipitate is
produced. The precipitate after recrystallisation from 5 ml of
1 ml of .0.1 M ceric ammonium sulphate is equivalent to ethanol (70 per cent) and drying at 105° melts at 226° to 231°
0.005585 g of Fe. (2.4.21).
Determine the weight per ml of the injection (2.4.29), and C. Melts at 170° to 174° (2.4.21).
calculate the percentage w/v of Fe.
For dextrans — Weigh accurately about 1.0 g, dilute to 500.0 Tests
ml with water, dilute 10.0 ml of this solution to 100.0 ml with Appearance of solution. A 5.0 per cent w/v solution in carbon
water, transfer 3.0 ml of the resulting solution to a test-tube dioxide-free water is clear (2.4.1), and not more intensely
and cool to 0°. Add, to form a lower layer, 6.0 ml of a solution coloured than reference solution BYS7 (2.4.1).
prepared and maintained at 0° containing 0.2 per cent w/v of
anthrone in a mixture of 19 volumes of sulphuric acid and 1 pH (2.4.24). 6.0 to 8.0, determined in a 5.0 per cent w/v solution.
volume of water, mix and immediately heat on a water-bath for Hydrazine. Determine by thin-layer chromatography (2.4.17),
5 minutes. Cool and measure the absorbance of the resulting coating the plate with silica gel GF254.
solution at about 625 nm (2.4.7). Repeat the operation using
Mobile phase. A mixture of 50 volumes of ethyl acetate, 20
3.0 ml of water in place of the dilution of the injection. From
volumes of acetone, 20 volumes of methanol and 10 volumes
the difference between the absorbances, calculate the content
of water.
of dextrose present using a calibration curve prepared by
treating suitable amounts of dextrose in the same manner. Test solution. Dissolve 1 g of the substance under examination
in sufficient of a mixture of equal volumes of acetone and
1 g of dextrose is equivalent to 0.94 g of dextrans.
water to produce 10 ml.
Determine the weight per ml of the injection (2.4.29), and
Reference solution. Dissolve 50 mg of hydrazine sulphate in
calculate the percentage w/v of dextrans.
50 ml of water and dilute to 100 ml with acetone; to 10 ml of
Labelling. The label states the strength in terms of the this solution add 0.2 ml of the test solution and dilute to 100 ml
equivalent amount of iron, Fe, in a suitable dose-volume. with a mixture of equal volumes of acetone and water.
629
ISONIAZID TABLETS IP 2007
After development, dry the plate in air and examine in Isoniazid Tablets
chromatogram obtained with the test solution is not more Isonicotinylhydrazid Tablets; INH Tablets
intense than the spot in the chromatogram obtained with the Isoniazid Tablets contain not less than 95.0 per cent and not
reference solution. Spray with dimethylaminobenzaldehyde more than 105.0 per cent of the stated amount of isoniazid,
reagent and examine in daylight. The additional spot (due to C6H7N3O.
hydrazine) in the chromatogram obtained with the reference
solution is more intense than any corresponding spot in the Identification
chromatogram obtained with the test solution.
A. Shake a quantity of the powdered tablets containing 0.1 g
Related substances. Determine by liquid chromatography of Isoniazid with 10 ml of ethanol (95 per cent) for 15 minutes,
(2.4.14). centrifuge and decant the supernatant liquid. Extract the residue
Test solution. Dissolve 50 mg of the substance under with two further quantities, each of 10 ml, of ethanol (95 per
examination in 100 ml of water. cent) and evaporate the combined extracts to dryness. The
residue complies with the following test.
octadecylsilyl silica gel (5 µm), Compare the spectrum with that obtained with isoniazid RS
– mobile phase: a mixture of 96 volumes of a solution or with the reference spectrum of isoniazid.
prepared by dissolving 1.4 g disodium hydrogen B. Shake a quantity of the powdered tablets containing 1 mg
phosphate and 1 ml of triethylamine to 1000 ml with of Isoniazid with 50 ml of ethanol (95 per cent) and filter. To
water, adjusting the pH to 6.0 with orthophosphoric 5 ml of the filtrate add 0.1 g of borax and 5 ml of a 5 per cent w/
acid and 4 volumes of acetonitrile, v solution of 1-chloro-2,4-dinitrobenzene in ethanol (95 per
– flow rate. 1 ml per minute, cent), evaporate to dryness on a water-bath and continue
– spectrophotometer set at 265 nm, heating for a further 10 minutes. To the residue add 10 ml of
– a 20 µl loop injector. methanol and mix; a reddish purple colour is produced.
Inject the test solution. Any individual impurity is not more
Tests
than 0.2 per cent and the sum of all impurities found is not
more than 1.0 per cent. Related substances. Determine by liquid chromatography
Heavy metals (2.3.13). 1.0 g complies with the limit test for (2.4.14).
heavy metals, Method B (20 ppm). Test solution. Weigh and powder 20 tablets. Weigh a quantity
Sulphated ash (2.3.18). Not more than 0.1 per cent. of the powdered tablets containing 50 mg of Isoniazid, dissolve
in 100 ml of the mobile phase.
on 1.0 g by drying in an oven at 105°. Use the chromatographic system described under Assay.
Assay. Determine by liquid chromatography (2.4.14). Inject the test solution. Any individual impurity is not more
than 1.0 per cent and the sum of all impurities found is not
Test solution. Dissolve about 25.0 mg of the substance under more than 2.0 per cent.
examination in 50.0 ml of water. Dilute 5.0 ml of this solution to
25.0 ml with water. Dissolution (2.5.2).
Reference solution. Dissolve 25.0 mg of the isoniazid RS in Apparatus. No 1
50.0 ml of water. Dilute 5.0 ml of this solution to 25.0 ml with Medium. 900 ml of water
water. Speed and time. 50 rpm and 45 minutes.
Use the chromatographic system described under the test for Withdraw a suitable volume of the medium and filter promptly
Related substances. through a membrane filter disc having an average pore diameter
not greater than 1.0 mµ, rejecting the first 1 ml of the filtrate.
Inject the reference solution. The tailing factor is not more Dilute suitably with water and measure the absorbance of the
than 2.0. The relative standard deviation for replicate injections resulting solution at the maximum at about 263 nm (2.4.7).
is not more than 2.0 per cent. Calculate the content of C6H7N3O taking 307 as the specific
Inject alternately the test solution and the reference solution. absorbance at 263 nm.
Calculate the content of C6H7N3O. D. Not less than 80 per cent of the stated amount of C6H7N3O.
Storage. Store protected from light. Other tests. Comply with the tests stated under Tablets.
630
IP 2007 ISOPRENALINE INJECTION
Assay. Determine by liquid chromatography (2.4.14). to brownish on standing exposed to air and almost immediately
Test solution. Weigh and powder 20 tablets. Weigh accurately after being made alkaline.
a quantity of the powder containing about 25.0 mg of Isoniazid
and dissolve in 50.0 ml of the mobile phase. Dilute 5.0 ml of
Identification
this solution to 25.0 ml with the mobile phase. Test A may be omitted if tests B, C and D are carried out. Tests
Reference solution. A 0.01 per cent w/v solution of the B and C may be omitted if tests A and D are carried out.
isoniazid RS in the mobile phase. A. Determine by infrared absorption spectrophotometry (2.4.6).
Chromatographic system Compare the spectrum with that obtained with isoprenaline
– a stainless steel column 15 cm x 4.6 mm, packed with hydrochloride RS or with the reference spectrum of
octadecylsilyl silica gel (5 µm) (such as Intersil ODS-3), isoprenaline hydrochloride.
– mobile phase: mix 96 volumes of a solution prepared by B. When examined in the range 230 nm to 360 nm (2.4.7), a
dissolving 1.4 g disodium hydrogen orthophosphate 0.005 per cent w/v solution shows an absorption maximum
anhydrous and 1.0 ml of triethylamine to 1000 ml with only at about 280 nm; absorbance at about 280 nm, about 0.5.
water and adjusting the pH to 6.0 with orthophosphoric
acid, and 4 volumes of acetonitrile and filter, C. To 2 ml of a freshly prepared 1 per cent w/v solution add
– flow rate. 1 ml per minute, 0.1 ml of ferric chloride test solution; an emerald-green colour
– spectrophotometer set at 265 nm, is produced which, on the gradual addition of sodium
– a 20 µl loop injector. bicarbonate solution, changes first to blue and then to red.
Inject the reference solution. The test is not valid unless the D. Gives the reactions of chlorides (2.3.1).
tailing factor is not more than 2.0 and the column efficiency is
not less than 2000 theoretical plates. The relative standard Tests
deviation for replicate injections is not more than 2.0 per cent. Isoprenalone. Absorbance of a 0.2 per cent w/v solution in
Inject alternately the test solution and the reference solution. 0.005 M sulphuric acid at about 310 nm, not more than
0.15 (2.4.7).
Calculate the content of C6H7N3O in the tablets.
Sulphates (2.3.17). Dissolve 0.5 g in 100 ml of water; 15 ml of
Storage. Store protected from light. the resulting solution complies with the limit test for sulphates
(0.2 per cent).
Isoprenaline Hydrochloride
Isoproterenol Hydrochloride on 1.0 g by drying over phosphorus pentoxide at a pressure
OH H Assay. Weigh accurately about 0.5 g, dissolve in 50 ml of
N CH3 anhydrous glacial acetic acid with the aid of the minimum of
,HCl
heat and titrate with 0.1 M perchloric acid, using crystal
CH3 violet solution as indicator. Carry out a blank titration.
HO
OH 1 ml of 0.1 M perchloric acid is equivalent to 0.02477 g of
C11H17NO3,HCl.
Isoprenaline Hydrochloride is (RS)-1-(3,4-dihydroxy-
phenyl)-2-isopropylaminoethanol hydrochloride.
Isoprenaline Hydrochloride contains not less than 98.0 per
cent and not more than 101.5 per cent of C11H17NO3,HCl, Isoprenaline Injection
Isoprenaline Hydrochloride Injection; Isoproterenol
Injection; Isoproterenol Hydrochloride Injection
almost odourless. Gradually darkens on exposure to air and
light; even in the absence of light, it is gradually degraded on Isoprenaline Injection is a sterile solution of Isoprenaline
exposure to a humid atmosphere, the decomposition being Hydrochloride in Water for Injections. It may contain suitable
faster at higher temperatures. Aqueous solutions become pink stabilising agents.
631
ISOPRENALINE SULPHATE IP 2007
Isoprenaline Hydrochloride Injection contains not less than Isoprenaline Sulphate

amount of isoprenaline hydrochloride, C11H17NO3,HCl. Isoproterenol Sulphate
Description. A colourless or very pale yellow solution.
OH H
Identification
N CH3
A. Determine by thin-layer chromatography (2.4.17), coating ,H2SO4, 2H2O
the plate with silica gel G. CH3
HO
Mobile phase. A mixture of 50 volumes of ethyl acetate, 30 OH 2
volumes of 2-propanol, 16 volumes of water and 4 volumes
(C11H17NO3)2,H2SO4,2H2O Mol. Wt. 247.7
Test solution. Use the injection diluted if necessary with
Isoprenaline Sulphate is (RS)-1-(3,4-dihydroxyphenyl)-2-
sufficient methanol (80 per cent) to produce a solution
isopropylaminoethanol sulphate dihydrate.
containing 0.02 per cent w/v of isoprenaline hydrochloride.
Isoprenaline Sulphate contains not less than 98.0 per cent
and not more than 102.0 per cent of (C11H17NO3)2,H2SO4,
isoprenaline hydrochloride RS in methanol (80 per cent).
detectable, place it for a few minutes in an atmosphere saturated
Identification
with diethylamine and spray with diazotised nitroaniline
solution. The chromatogram obtained with the test solution Test A may be omitted if tests B, C and D are carried out. Tests
exhibits an elongated zone corresponding to that obtained B and C may be omitted if tests A and D are carried out.
A. Dissolve 0.5 g in 1.5 ml of water, add 3.5 ml of 2-propanol
B. To 2 ml add 0.1 ml of ferric chloride test solution; an emerald- scratch the walls of the container with a glass rod to induce
green colour develops which, on gradual addition of sodium crystallisation, collect the crystals and dry over phosphorus
bicarbonate solution, changes to blue and then to red. pentoxide at 60° at a pressure of 1.5 to 2.5 kPa. The crystals
comply with the following test.
Tests
pH (2.4.24). 2.5 to 3.0. Compare the spectrum with that obtained with isoprenaline
Other tests. Complies with the tests stated under Parenteral sulphate RS treated in the same manner.
Preparations (Injections). B. To 1 ml of a 1 per cent w/v solution add 0.1 ml of ferric
Assay. To an accurately measured volume containing about chloride test solution; an emerald-green colour is produced
5 mg of Isoprenaline Hydrochloride add sufficient water to which, on gradual addition of sodium bicarbonate solution,
produce 50.0 ml. To 20.0 ml add 0.5 ml of ferrous sulphate- changes first to blue and then to red.
citrate solution and 2 ml of glycine buffer solution, mix and C. To 5 ml of a freshly prepared 1 per cent w/v solution add
allow to stand for 20 minutes. Add sufficient water to produce 0.15 ml of silver nitrate solution; a greyish precipitate is
25.0 ml, mix and measure the absorbance of the resulting produced on standing for 10 minutes and the solution becomes
solution at the maximum at about 540 nm (2.4.7). Calculate the pink.
content of C11H17NO3,HCl from the absorbance obtained by
D. A 10 per cent w/v solution gives the reaction of sulphates
repeating the determination using 2.0 ml of a 0.1 per cent w/v
(2.3.1).
solution of isoprenaline hydrochloride RS in place of the
Tests
exceeding 30°. Appearance of solution. A freshly prepared 10.0 per cent w/v
solution in carbon dioxide-free water is clear (2.4.1), and not
more intensely coloured than reference solution YS6 (2.4.1).
pH (2.4.24). 4.0 to 5.5, determined in a freshly prepared 1.0 per
cent w/v solution in carbon dioxide-free water.
632
IP 2007 ISOPROPYL ALCOHOL
Isoprenalone. Absorbance of a 0.2 per cent w/v solution in the absorbance of the resulting solution at the maximum at
0.005 M sulphuric acid at about 310 nm, not more than about 540 nm (2.4.7). Calculate the content of
0.2 (2.4.7). (C11H17NO3)2,H2SO4,2H2O in the tablet from the absorbance
Heavy metals (2.3.13). 2.0 g complies with the limit test for obtained by repeating the determination using 2.0 ml of a
heavy metals, Method A (10 ppm). 0.1 per cent w/v solution of isoprenaline sulphate RS in place
of the substance under examination.
Water (2.3.43). 5.0 to 7.5, determined on 0.2 g.
Assay. Weigh accurately about 0.8 g, dissolve in 40 ml of quantity of the powder containing about 0.1 g of Isoprenaline
anhydrous glacial acetic acid, warming gently if necessary Sulphate and shake with 50 ml of water for 15 minutes. Add
and titrate with 0.1 M perchloric acid, using crystal violet sufficient water to produce 100.0 ml, mix and filter. Dilute 20.0
solution as indicator. Carry out a blank titration. ml of the filtrate to 200.0 ml with water. To 20.0 ml of the
1 ml of 0.1 M perchloric acid is equivalent to 0.05206 g of resulting solution add 0.5 ml of ferrous sulphate-citrate
(C11H17NO3)2,H2SO4. solution and 2 ml of glycine buffer solution and allow to stand
for 20 minutes. Dilute to 25.0 ml with water and mix. Measure
Storage. Store protected from light. the absorbance of the resulting solution at the maximum at
about 540 nm (2.4.7). Calculate the content of
(C11H17NO3)2,H2SO4,2H2O from the absorbance obtained by
Isoprenaline Tablets repeating the determination using 2.0 ml of a 0.1 per cent w/v
solution of isoprenaline sulphate RS in place of the substance
Isoprenaline Sulphate Tablets; Isoproterenol Tablets; under examination.
Isoproterenol Sulphate Tablets
Isoprenaline Tablets contain not less than 90.0 per cent and
isoprenaline sulphate, (C11H17NO3)2,H2SO4,2H2O.
Isopropyl Alcohol
Identification
2-Propanol; Propan-2-ol
A. Extract a quantity of the powdered tablets containing about
H OH
50 mg of Isoprenaline Sulphate with 5 ml of water and filter.
Reserve the filtrate for test C. To 1 ml of the filtrate add 0.1 ml H3C CH3
of ferric chloride test solution; an emerald-green colour is
produced which, on gradual addition of sodium bicarbonate C3H8O Mol. Wt. 60.1
solution, changes first to blue and then to red. Isopropyl Alcohol is propan-2-ol.
B. Extract a quantity of the powdered tablets containing about Description. A clear, colourless liquid; odour, characteristic
50 mg of Isoprenaline Sulphate with 5 ml of water and filter. To and spirituous; flammable.
the filtrate add 0.15 ml of silver nitrate solution; a greyish
precipitate is produced on standing for 10 minutes and the Identification
solution becomes pink. A. Mix 1 ml of a 10 per cent v/v solution with 2 ml of mercuric
C. To 2 ml of the filtrate reserved from test A add 0.5 ml of sulphate solution and heat just to boiling; a white or yellowish
dilute hydrochloric acid and 0.5 ml of barium chloride white precipitate is produced.
solution; a white precipitate is formed. B. Gently heat 1 ml with 4 ml of dilute potassium dichromate
Tests solution and 1 ml of sulphuric acid; acetone, recognisable by
its odour, is evolved.
Tablets. Tests
Crush one tablet and shake with 50 ml of water for 15 minutes. Acidity or alkalinity. Gently boil 25 ml for 5 minutes with 25 ml
Add sufficient water to produce 100.0 ml, mix and filter. To of carbon dioxide-free water and cool, taking precautions to
20.0 ml of the filtrate add 0.5 ml of ferrous sulphate-citrate exclude carbon dioxide. Not more than 0.06 ml of 0.1 M sodium
solution and 2 ml of glycine buffer solution and allow to stand hydroxide is required to make the resulting solution alkaline
for 20 minutes. Dilute to 25.0 ml with water and mix. Measure to phenolphthalein solution.
633
DILUTED ISOSORBIDE DINITRATE IP 2007
Distillation range (2.4.8). Not less than 95.0 per cent v/v distils In the chromatogram obtained with reference solution (a) the
between 81° and 83°. sum of areas of any peaks other than the principal peak and
Refractive index (2.4.27). 1.377 to 1.378, determined at 20°. the peaks due to 2-butanol is not greater than 3 times the area
of the peak due to 2-butanol (0.3 per cent).
Weight per ml (2.4.29). 0.782 g to 0.786 g, determined at 20°.
Non-volatile matter. Not more than 0.002 per cent w/v,
Aldehydes and ketones. Mix in a cylinder 25 ml with 25 ml of determined by evaporating 100 ml on a water-bath and drying
water and 50 ml of hydroxylamine solution, allow to stand for the residue at 105°.
5 minutes and titrate with 0.1 M sodium hydroxide until the
colour is the same as that of a mixture of 50 ml of hydroxylamine Water-insoluble matter. Mix 1 volume with 19 volumes of
solution and 50 ml of water contained in a similar cylinder, water; no opalescence is produced.
each being viewed down the vertical axis of the cylinder. Not Water (2.3.43). Not more than 0.5 per cent, determined on 5 g.
more than 2.0 ml of 0.1 M sodium hydroxide is required.
Benzene and related substances. Determine by gas
Test solution. The substance under examination.
Diluted Isosorbide Dinitrate
Reference solution (a). A 0.1 per cent v/v solution of 2- Diluted Sorbide Dinitrate; Diluted Sorbide Nitrate
butanol reagent in the substance under examination.
Reference solution (b). A solution containing 0.1 per cent v/v H ONO2
of each of 2-butanol reagent and 1-propanol in the substance O
under examination.
O
Reference solution (c). A 0.0002 per cent v/v solution of H
benzene in the substance under examination. O2NO
Chromatographic system C6H8N2O8 Mol. Wt. 236.1

diatomaceous support (80 to 100 mesh) coated with 15 Diluted Isosorbide Dinitrate is a dry mixture of 1,4:3,6-
per cent w/w of polyethylene glycol 400, dianhydro-D-glucitol 2,5-dinitrate with Lactose, Mannitol or
– temperature: other suitable inert diluent. It may contain a suitable stabilising
column.50°, agent.
inlet port.150°, Diluted Isosorbide Dinitrate contains not less than 95.0 per
– flow rate. 30 ml per minute of the carrier gas, cent and not more than 105.0 per cent of the stated amount of
– flame ionisation detector at 200°. isosorbide dinitrate, C6H8N2O8 and usually contains 20 per
Inject separately 2 µl of each of the test solution and reference cent to 50 per cent of isosorbide dinitrate.
solution (a). The chromatogram obtained with the test solution Description. A fine, white, crystalline powder; odourless or
shows no peak with retention time similar to the peak due to 2- almost odourless.
butanol (retention time relative to isopropyl alcohol, about
1.5) obtained with solution (2). Inject 2 µl of reference solution CAUTION — Undiluted isosorbide dinitrate is a powerful
(b) and adjust the sensitivity of the system so that the heights explosive and can be exploded with percussion or excessive
of the peaks due to 2-butanol and 1-propanol in the heat. Proper precautions must be taken in handling it and
chromatogram obtained with reference solution (b) are not only exceedingly small amounts should be isolated.
less than 50 per cent of the full scale of the recorder. The test Identification
is not valid unless the resolution between the peaks due to 2-
butanol and 1-propanol in the chromatogram obtained with A. Shake a quantity containing 50 mg of isosorbide dinitrate
reference solution (b) is at least 1.2. with 15 ml of acetone for 2 minutes. Filter, evaporate the filtrate
Inject alternately 2 µl each of the test solution and reference to dryness at a temperature not exceeding 35° and dry the
solution (c). The area of any peak due to benzene in the residue over phosphorus pentoxide at a pressure of 0.7 kPa
chromatogram obtained with the test solution is not greater for 16 hours. The residue complies with the following test.
than the difference between the area of the peak due to benzene Determine by infrared absorption spectrophotometry (2.4.6).
in the chromatogram obtained with reference solution (c) and Compare the spectrum with that obtained with diluted
that of the peak due to benzene in the chromatogram obtained isosorbide dinitrate RS treated in the same manner or with
with the test solution. the reference spectrum of isosorbide dinitrate.
634
IP 2007 DILUTED ISOSORBIDE DINITRATE
B. Determine by thin-layer chromatography (2.4.17), coating chromatogram obtained with reference solution (c) is not less
the plate with silica gel G. than 20 per cent of the full scale of the recorder.
Mobile phase. Toluene. Inject 10 µl of reference solution (e). The test is not valid
unless in the chromatogram obtained with reference solution
Test solution. Extract a quantity containing 2 mg of isosorbide
(e), the resolution between the peaks corresponding to
dinitrate with 1 ml of ether and centrifuge.
isosorbide dinitrate and isosorbide 2-nitrate is at least 6.0.
Inject 10 µl of test solution (a), 10 µl of reference solution (c)
solution but using diluted isosorbide dinitrate RS in place of
and 10 µl of reference solution (d). In the chromatogram
the substance under examination.
obtained with test solution (a) the area of any peak
Apply to the plate 20 µl of each solution. After development, corresponding to isosorbide 2-nitrate is not greater than the
dry the plate in a current of air, spray with a 1 per cent w/v area of the principal peak in the chromatogram obtained with
solution of diphenylamine in methanol and expose for reference solution (c) (0.5 per cent); the area of any peak
15 minutes to ultraviolet light at 254 and 365 nm. The principal corresponding to isosorbide 5-nitrate is not greater than the
spot in the chromatogram obtained with the test solution area of the principal peak in the chromatogram obtained with
corresponds to that in the chromatogram obtained with the reference solution (d) (0.5 per cent).
reference solution.
C. Extract a quantity containing 10 mg of isosorbide dinitrate heavy metals, Method B (10 ppm).
with 10 ml of ether and filter. Evaporate the filtrate to dryness
at a temperature not exceeding 35° and dissolve the residue in
0.15 ml of sulphuric acid (50 per cent) containing a trace of
diphenylamine; an intense blue colour is produced.
Tests Test solution (a). Mix an accurately weighed quantity of the
Inorganic nitrates. Determine by thin-layer chromatography substance under examination containing about 25 mg of
(2.4.17), coating the plate with silica gel H. isosorbide dinitrate with 20 ml of the mobile phase with the aid
of ultrasound for 15 minutes and dilute to 25.0 ml with the
Mobile phase. A mixture of 60 volumes of toluene, 30 volumes mobile phase. Filter the solution through a suitable membrane
of acetone and 15 volumes of glacial acetic acid. filter.
Test solution. A solution of the substance under examination Test solution (b). Dilute 1.0 ml of test solution (a) to 10.0 ml
in ethanol (95 per cent) containing the equivalent of 2.0 per with the mobile phase.
cent w/v of isosorbide dinitrate.
Reference solution (a). Mix a quantity of isosorbide dinitrate
Reference solution. Prepare freshly a 0.01 per cent w/v solution RS containing 25.0 mg of isosorbide dinitrate with 20 ml of the
of potassium nitrate in ethanol (90 per cent). mobile phase with the aid of ultrasound for 15 minutes and
Apply to the plate 10 µl of each solution. After development, dilute to 25.0 ml with the mobile phase. Filter the solution
dry the plate in a current of air until the acetic acid is completely through a suitable membrane filter.
removed. Spray copiously with freshly prepared potassium Reference solution (b). Dilute 1.0 ml of reference solution (a)
iodide and starch solution. Expose the plate to ultraviolet to 10.0 ml with the mobile phase.
light at 254 nm for 15 minutes. Examine in daylight. Any spot
Reference solution (c). Dissolve 10.0 mg of isosorbide 2-
corresponding to potassium nitrate in the chromatogram
nitrate RS in the mobile phase and dilute to 10.0 ml with the
mobile phase. Dilute 0.1 ml of this solution to 20.0 ml with the
mobile phase.
Reference solution (d). Dissolve 20.0 mg of isosorbide
(2.4.14) by the method described under Assay, changing the
mononitrate RS in the mobile phase and dilute to 10.0 ml with
detector setting to 210 to 215 nm and using a 10 µl injector.
the mobile phase. Dilute 0.1 ml of this solution to 20.0 ml with
Under the stated conditions, the retention times are: isosorbide the mobile phase.
dinitrate, about 5 minutes; isosorbide 2-nitrate, about 8
Reference solution (e). Dissolve 5 mg of isosorbide 2-nitrate
minutes; isosorbide 5-nitrate, about 11 minutes.
RS in the mobile phase and dilute to 10 ml with the mobile
Inject 10 µl of reference solution (c). Adjust the sensitivity of phase. To 1 ml of this solution add 0.5 ml of reference solution
the system so that the height of the principal peak in the (a) and dilute to 10 ml with the mobile phase.
635
ISOSORBIDE DINITRATE TABLETS IP 2007
Chromatographic system Apply to the plate 20 µl of each solution. After development,

– a stainless steel column 25 cm x 4.6 mm packed with dry the plate in a current of air, spray with a 1 per cent w/v
aminopropylmethylsilyl silica gel (10 µm), solution of diphenylamine in methanol and expose for 15
– mobile phase: a mixture of 15 volumes of ethanol and 85 minutes to ultraviolet light at 254 nm and 365 nm. The principal
volumes of trimethylpentane, spot in the chromatogram obtained with the test solution
– flow rate. 1 ml per minute, corresponds to that in the chromatogram obtained with the
– spectrophotometer set at 230 nm, reference solutionB. Extract a quantity of the powdered tablets
– a 20 µl loop injector. containing10mgoisosorbide dinitrate with 10 ml of ether and
Inject reference solution (b). Adjust the sensitivity of the filter. Evaporate the filtrate to dryness at a temperature not
system so that the height of the principal peak in the exceeding 35° and dissolve the residue in 0.15 ml of sulphuric
chromatogram obtained is not less than 50 per cent of the full acid (50 per cent) containing a trace of diphenylamine; an
scale of the recorder. If the areas of the peaks from two intense blue colour is produced.
successive injections do not agree to within 1.0 per cent, then
inject a further four times and calculate, for the six injections,
Tests
the relative standard deviation. The assay is not valid unless Inorganic nitrates. Determine by thin-layer chromatography
the relative standard deviation for the six injections is at most (2.4.17), coating the plate with silica gel H.
2.0 per cent.
Inject alternately test solution (b) and reference solution (b). of acetone and 15 volumes of glacial acetic acid.
Calculate the content of isosorbide dinitrate as a percentage Test solution. Shake a quantity of the powdered tablets
of the declared content. containing 0.1 g of isosorbide nitrate with 5 ml of ethanol
(95 per cent) and filter.
Storage. Store protected from light, at a temperature not
exceeding 15°. Reference solution. Prepare freshly a 0.01 per cent w/v solution
of potassium nitrate in ethanol (90 per cent).
Labelling. The label states the percentage content of
isosorbide dinitrate. Apply to the plate 10 µl of each solution. After development,
dry the plate in a current of air until the acetic acid is completely
removed. Spray copiously with freshly prepared potassium
iodide and starch solution. Expose the plate to ultraviolet
light at 254 nm for 15 minutes. Examine in daylight. Any spot
Isosorbide Dinitrate Tablets corresponding to potassium nitrate in the chromatogram
Sorbide Dinitrate Tablets; Sorbide Nitrate Tablets spot in the chromatogram obtained with the reference solution.
Isosorbide Dinitrate Tablets contain not less than 90.0 per Related substances. Determine by liquid chromatography
cent and not more than 110.0 per cent of the stated amount of (2.4.14).
isosorbide dinitrate, C6H8N2O8.
Test solution. Add 20 ml of the mobile phase to a quantity of
Identification the powdered tablets containing 25 mg of isosorbide dinitrate,
mix with the aid of ultrasound for 15 minutes and dilute to 25
CAUTION - Undiluted isosorbide dinitrate is a powerful ml with the same solvent. Filter through a glass-fibre filter
explosive and can be exploded with percussion or excessive (such as Whatman GF/C) and use the filtrate.
heat. Proper precautions must be taken in handling it and
only exceedingly small amounts should be isolated. Reference solution (a). A solution containing 0.0005 per cent
w/v of isosorbide 2-nitrate RS in the mobile phase.
the plate with silica gel G. Reference solution (b). A solution containing 0.0005 per cent
w/v of isosorbide mononitrate RS in the mobile phase.
Mobile phase. Toluene.
Test solution. Extract a quantity of the powdered tablets w/v each of isosorbide dinitrate RS and isosorbide 2-nitrate
containing 2 mg of isosorbide dinitrate with 1 ml of ether and RS in the mobile phase.
centrifuge.
Reference solution. Dissolve 2 mg of isosorbide dinitrate RS – a stainless steel column 25 cm x 4.6 mm packed with
in 1 ml of ether. aminopropylmethylsilyl silica gel (10 µm),
636
IP 2007 ISOSORBIDE DINITRATE TABLETS
– mobile phase: a mixture of 15 volumes of ethanol and 85 15 minutes, add 25 ml of water, make alkaline with strong
volumes of 2,2,4-trimethylpentane, ammonia solution, cool and add sufficient water to produce
– flow rate. 1 ml per minute, 50.0 ml. Measure the absorbance of the resulting solution at
– spectrophotometer set at 215 nm, about 405 nm (2.4.7), using as the blank 1.0 ml of glacial
– a 20 µl loop injector. acetic acid treated in a similar manner beginning at the words
Inject reference solution (c). The test is not valid unless, in “add 2 ml of phenoldisulphonic acid solution,....”. Dissolve
the chromatogram obtained, the resolution between the peaks 0.2 g of potassium nitrate, previously dried at 105°, in 5 ml of
corresponding to isosorbide dinitrate and isosorbide 2-nitrate water and add sufficient glacial acetic acid to produce 25.0
is at least 6.0. ml. To 5.0 ml add sufficient glacial acetic acid to produce 50.0
ml. Using 1.0 ml of this solution repeat the procedure beginning
In the chromatogram obtained with the test solution the area at the words “add 2 ml of phenoldisulphonic acid
of any peak corresponding to isosorbide 2-nitrate is not greater
solution,......”. Calculate the content of C6H8N2O8 from the
values of the absorbances so obtained.
obtained with reference solution (a) (0.5 per cent) and the area
of any peak corresponding to isosorbide 5-nitrate is not greater 1 ml of the potassium nitrate solution is equivalent to 0.000934
than the area of the principal peak obtained with reference g of C6H8N2O8.
solution (b) (0.5 per cent).
Calculate the content of C6H8N2O8 in the tablet.
Dissolution (2.5.2). (for tablets intended to be swallowed
whole). Other tests. Comply with the tests stated under Tablets.
Apparatus. No 1 Assay. Determine by liquid chromatography (2.4.14).

Medium. 900 ml of 0.1 M hydrochloric acid Test solution. Weigh and powder 20 tablets. Add 20 ml of the
Speed and time. 50 rpm and 30 minutes. mobile phase to an accurately weighed quantity of the
Withdraw 10 ml of the medium and filter promptly through a powdered tablets containing about 25 mg of isosorbide
membrane filter disc having an average pore diameter not dinitrate, mix with the aid of ultrasound for 15 minutes and
greater than 1.0 µm, rejecting the first 1 ml of the filtrate. dilute to 25.0 ml with the same solvent. Filter through a glass-
fibre filter (such as Whatman GF/C) and dilute 1 volume to 10
Determine by liquid chromatography (2.4.14). volumes with the mobile phase.
Test solution. The filtrate from the dissolution medium. Reference solution (a). Add 20 ml of the mobile phase to a
Reference solution. A solution of isosorbide dinitrate RS in quantity of isosorbide dinitrate RS containing 25 mg of
the dissolution medium containing the same concentration of isosorbide dinitrate, mix with the aid of ultrasound for 15
isosorbide dinitrate as that expected in the dissolution medium minutes and dilute to 25 ml with the mobile phase. Filter through
in the vessel. a glass-fibre filter (such as Whatman GF/C) and dilute 1 volume
to 10 volumes with the mobile phase.
– a stainless steel column 20 cm x 4.6 mm packed with Reference solution (b). A solution containing 0.005 per cent
octadecylsilyl silica gel (5 µm) (such as Hypersil ODS), w/v each of isosorbide dinitrate RS and isosorbide 2-nitrate
– mobile phase: a mixture of equal volumes of methanol RS in the mobile phase.
and water,
Follow the procedure described under Related substances
but using a detection wavelength of 230 nm.
– spectrophotometer set at 222 nm.
– a 100 µl loop injector. The test is not valid unless, in the chromatogram obtained
Calculate the content of C6H8N2O8. with reference solution (b), the resolution between the peaks
corresponding to isosorbide dinitrate and isosorbide 2-nitrate
D. Not less than 80 per cent of the stated amount of C6H8N2O8. is at least 6.0.
Uniformity of content. Comply with the test stated under Calculate the content of C6H8N2O8 in the tablets from the
Tablets.
chromatograms obtained.
Crush one tablet, add 5 ml of glacial acetic acid, shake for
Storage Store at a temperature not exceeding 30°.
1 hour and centrifuge. To a suitable volume of the supernatant
liquid containing 1.0 mg of isosorbide dinitrate add sufficient Labelling. The label states whether the tablets are to be
glacial acetic acid, if necessary, to produce 1.0 ml, add 2 ml of swallowed whole, chewed before swallowing or allowed to
phenoldisulphonic acid solution, allow to stand for dissolve in the mouth.
637
ISOXSUPRINE HYDROCHLORIDE IP 2007
Isoxsuprine Hydrochloride using 1-naphtholbenzein solution as indicator. Carry out a

blank titration.
OH H C18H23NO3,HCl.
N ,HCl
O
CH3 CH3
HO
Isoxsuprine Injection
Isoxsuprine Hydrochloride Injection
Isoxsuprine Hydrochloride is (1RS,2SR)-1-(4-
hydroxyphenyl)-2-[(1RS)-1-methyl-2- Isoxsuprine Injection is a sterile solution of Isoxsuprine
phenoxyethylamino]propan-1-ol hydrochloride. Hydrochloride in Water for Injections.
Isoxsuprine Hydrochloride contains not less than 98.5 per Isoxsuprine Injection contains not less than 95.0 per cent and
cent and not more than 101.0 per cent of C18H23NO3,HCl, not more than 105.0 per cent of the stated amount of
calculated on the dried basis. isoxsuprine hydrochloride, C18H23NO3,HCl.
Description. A white or almost white, crystalline powder; Identification

A. To a volume containing 50 mg of Isoxsuprine Hydrochloride
Identification add 20 ml of water and 10 ml of ammonia buffer pH 10.0 and
extract with three quantities, each of 15 ml, of dichloromethane.
A. Determine by infrared absorption spectrophotometry (2.4.6). Shake the combined extracts with 5 g of anhydrous sodium
Compare the spectrum with that obtained with isoxsuprine sulphate, filter, evaporate the filtrate to dryness, dissolve the
hydrochloride RS or with the reference spectrum of residue in 5 ml of 0.1 M methanolic hydrochloric acid and
isoxsuprine hydrochloride. evaporate to dryness. Dissolve the residue in 5 ml of methanol,
B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.01 evaporate to dryness, redissolve the residue in 2 ml of
per cent w/v solution in 0.1 M hydrochloric acid shows methanol, add 15 ml of dichloromethane, again evaporate to
absorption maxima at about 269 nm and 274 nm; absorbance dryness and dry the residue at 60° at a pressure of 2 kPa for 1
at about 269 nm, about 0.73 and at about 274 nm, about 0.72. hour.
C. Dissolve 10 mg in 1 ml of water and add 0.05 ml of copper On the residue determine by infrared absorption
sulphate solution and 1 ml of 5 M sodium hydroxide; a blue spectrophotometry (2.4.6). Compare the spectrum with that
colour is produced. Add 1 ml of ether and shake; the ether obtained with isoxsuprine hydrochloride RS treated in the
layer remains colourless. same manner or with the reference spectrum of isoxsuprine
hydrochloride.
Tests final solution obtained in the Assay shows absorption maxima
at about 269 nm and 274 nm.
pH (2.4.24). 4.5 to 6.0, determined in a 1.0 per cent w/v solution,
prepared with gentle warming if necessary. Tests
Phenones. Absorbance of a 0.01 per cent w/v solution at about
pH (2.4.24). 4.9 to 6.0.
310 nm, not more than 0.20 (2.4.7).
Heavy metals (2.3.13). 1.0 g complies with the limit test for Other tests. Complies with the tests stated under Parenteral
heavy metals, Method B (20 ppm). Preparations (Injections).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Assay. To an accurately measured volume containing about
50 mg of Isoxsuprine Hydrochloride add sufficient 0.1 M
Loss on drying (2.4.19). Not more than 0.5 per cent, determined hydrochloric acid to produce 100.0 ml. Dilute 10.0 ml to 100.0
on 1.0 g by drying in an oven at 105°. ml with the same solvent and measure the absorbance of the
Assay. Weigh accurately about 0.6 g, dissolve in 150 ml of resulting solution at the maximum at about 274 nm (2.4.7).
anhydrous glacial acetic acid, heating on a water-bath to Calculate the content of C18H23NO3,HCl taking 73 as the specific
effect dissolution. Cool and titrate with 0.1 M perchloric acid, absorbance at 274 nm.
638
IP 2007 ISOXSUPRINE TABLETS
Isoxsuprine Tablets On the residue determine by infrared absorption

Isoxsuprine Hydrochloride Tablets obtained with isoxsuprine hydrochloride RS treated in the
Isoxsuprine Tablets contain not less than 92.5 per cent and same manner or with the reference spectrum of isoxsuprine
not more than 107.5 per cent of the stated amount of hydrochloride.
isoxsuprine hydrochloride, C18H23NO3,HCl. B. When examined in the range 230 nm to 360 nm (2.4.7), the
final solution obtained in the Assay shows absorption maxima
Identification at about 269 nm and 274 nm.
A. To a quantity of the powdered tablets containing 50 mg of
Tests
Isoxsuprine Hydrochloride add 50 ml of 0.1 M hydrochloric
acid and heat on a water-bath for 30 minutes. Cool, filter, add Other tests. Comply with the tests stated under Tablets.
10 ml of ammonia buffer pH 10.0 and extract with three
quantities, each of 15 ml, of dichloromethane. Shake the
quantity of the powder containing about 20 mg of Isoxsuprine
combined extracts with 5 g of anhydrous sodium sulphate,
Hydrochloride, add 50 ml of 0.1 M hydrochloric acid and boil
filter, evaporate the filtrate to dryness, dissolve the residue in
on a water-bath for 30 minutes. Cool, add sufficient 0.1 M
5 ml of 0.1 M methanolic hydrochloric acid and evaporate to
hydrochloric acid to produce 100 0 ml, mix and filter. Dilute
dryness. Dissolve the residue in 5 ml of methanol, evaporate
25.0 ml of the filtrate to 100.0 ml with 0.1 M hydrochloric acid
to dryness, redissolve the residue in 2 ml of methanol, add 15
ml of dichloromethane, again evaporate to dryness and dry
the residue at 60° at a pressure of 2 kPa for 1 hour.
C18H23NO3,HCl taking 73 as the specific absorbance at 274 nm.
639
K
Kanamycin Sulphate ....
Kanamycin Acid Sulphate ....
Kanamycin Injection ....
Heavy Kaolin ....
Light Kaolin ....
Ketamine Hydrochloride ....
Ketamine Injection ....
Ketoconazole ....
Ketoconazole Tablets ....
Ketoprofen ....
Ketoprofen Capsules ....
641
IP 2007 KANAMYCIN SULPHATE
spray with a mixture of equal volumes of a 0.2 per cent w/v

Kanamycin Sulphate solution of 1,3-napthalenediol in ethanol (95 per cent) and
a 45 per cent w/v solution of sulphuric acid and heat at 150°
H2 N for 5 to 10 minutes. The principal spot in the chromatogram
O chromatogram obtained with reference solution (a). The test
H2N NH2 is not valid unless the chromatogram obtained with reference
OH solution (b) shows three clearly separated spots.
O B. Dissolve 0.5 g in 10 ml of water and add 10 ml of a 1 per cent
OH
OH w/v solution of picric acid. If necessary initiate crystallisation
OH by scratching the walls of the container with a glass rod, allow
OH , H2SO4, H2O to stand and filter. The crystals, after washing with 20 ml of
O water and drying at 105°, melt at about 235°, with
O
decomposition (2.4.21).
OH
C. Dissolve 50 mg in 2 ml of water, add 1 ml of a 1 per cent w/
NH2 v solution of ninhydrin and heat for a few minutes on a water-
NH2 bath; a violet colour is produced.
D. Gives the reactions of sulphates (2.3.1).
C18H36N4O11,H2SO4,H2O Mol. Wt. 600.6
Kanamycin Sulphate is 6-O-(3-amino-3-deoxy-α-D-
Tests
glucopyranosyl)-4-O-(6-amino-6-deoxy-α-D- pH (2.4.24). 6.5 to 8.5, determined in a 1.0 per cent w/v solution.
glucopyranosyl)-2-deoxystreptamine sulphate
monohydrate, an antimicrobial substance produced by the Specific optical rotation (2.4.22). +112° to +123°, determined
growth of certain strains of Streptomyces kanamyceticus. at 20° in a 1.0 per cent w/v solution.
Kanamycin Sulphate has a potency of not less than 750 Units Kanamycin B. Determine by thin-layer chromatography
per mg, calculated on the dried basis. (2.4.17), coating the plate with a 0.75-mm layer of the following
mixture. Mix 0.3 g of carbomer (such as Carbopol) with 240 ml
Description. A white or almost white, crystalline powder; of water and allow to stand, with moderate shaking, for
odourless or almost odourless. 1 hour; adjust to pH 7.0 by the gradual addition, with
continuous shaking, of 2 M sodium hydroxide and add 30 g
Identification of silica gel H. Heat the plate at 110° for 1 hour, allow to cool
A. Determine by thin-layer chromatography (2.4.17), coating and use immediately.
the plate with a 0.75-mm layer of the following mixture. Mix Mobile phase. A 7 per cent w/v solution of potassium
0.3 g of carbomer (such as Carbopol 934) with 240 ml of water dihydrogen phosphate.
and allow to stand, with moderate shaking, for 1 hour; adjust
to pH 7.0 by the gradual addition, with continuous shaking, of Test solution. A 0.5 per cent w/v solution of the substance
2 M sodium hydroxide and add 30 g of silica gel H. Heat the under examination in water.
plate at 110° for 1 hour, allow to cool and use immediately. Reference solution. A 0.02 per cent w/v solution of kanamycin
Mobile phase. A 7 per cent w/v solution of potassium B sulphate RS in water.
dihydrogen phosphate. Apply to the plate 4 µl of each solution. Allow the mobile
Test solution. A 0.1 per cent w/v solution of the substance phase to rise 12 cm. Dry the plate in a current of warm air,
under examination in water. spray with ninhydrin and stannous chloride reagent and
heat at 110° for 15 minutes. Any spot corresponding to
Reference solution (a). A 0.1 per cent w/v solution of kanamycin B in the chromatogram obtained with the test
kanamycin sulphate RS in water. solution is not more intense than the spot in the chromatogram
Reference solution (b). A solution containing 0.1 per cent w/ obtained with the reference solution.
v each of kanamycin sulphate RS, neomycin sulphate RS and Sulphates. 15.0 to 17.0 per cent of SO4, calculated on the dried
streptomycin sulphate RS in water. basis and determined by the following method. Dissolve 0.25
Apply to the plate 10 µl of each solution. Allow the mobile g in 100 ml of water and adjust the pH to 11 using strong
phase to rise 12 cm. Dry the plate in a current of warm air, ammonia solution. Add 10.0 ml of 0.1 M barium chloride and
643
KANAMYCIN ACID SULPHATE IP 2007
0.5 mg of metalphthalein. Titrate with 0.1 M disodium and allow to stand, with moderate shaking, for 1 hour; adjust
edetate; when the colour of the solution begins to change, to pH 7.0 by the gradual addition, with continuous shaking, of
add 50 ml of ethanol (95 per cent) and continue the titration 2 M sodium hydroxide and add 30 g of silica gel H. Heat the
until the blue colour disappears. plate at 110° for 1 hour, allow to cool and use immediately.
1 ml of 0.1 M barium chloride is equivalent to 0.009606 g of Mobile phase. A 7 per cent w/v solution of potassium
SO4. dihydrogen phosphate.
Sulphated ash (2.3.18). Not more than 0.5 per cent. Test solution. A 0.1 per cent w/v solution of the substance
on 1.0 g by drying in an oven at 60° at a pressure not exceeding Reference solution (a). A 0.1 per cent w/v solution of
0.7 kPa for 3 hours. kanamycin sulphate RS in water.
Assay. Determine by the microbiological assay of antibiotics, Reference solution (b). A solution containing 0.1 per cent w/
Method A or B (2.2.10). v each of kanamycin sulphate RS, neomycin sulphate RS and
streptomycin sulphate RS in water.
Kanamycin Sulphate intended for use in the manufacture of
parenteral preparations without a further appropriate Apply to the plate 10 µl of each solution. Allow the mobile
procedure for the removal of bacterial endotoxins complies phase to rise 12 cm. Dry the plate in a current of warm air,
with the following additional requirement. spray with a mixture of equal volumes of a 0.2 per cent w/v
solution of 1,3-napthalenediol in ethanol (95 per cent) and
Bacterial endotoxins (2.2.3). Not more than 0.67 Endotoxin a 45 per cent w/v of solution of sulphuric acid and heat at
Unit per mg of kanamycin. 150° for 5 to 10 minutes. The principal spot in the chromatogram
Kanamycin Sulphate intended for use in the manufacture of obtained with the test solution corresponds to that in the
parenteral preparations without a further appropriate chromatogram obtained with reference solution (a). The test
sterilisation procedure complies with the following is not valid unless the chromatogram obtained with reference
additional requirement. solution (b) shows three clearly separated spots.
Sterility (2.2.11). Complies with the test for sterility. B. Dissolve 0.5 g in 10 ml of water and add 10 ml of a 1 per cent
w/v solution of picric acid. If necessary initiate crystallisation
Storage. Store protected from light and moisture. If the by scratching the walls of the container with a glass rod, allow
contents are intended for use in the manufacture of parenteral to stand and filter. The melting point of the crystals, after
preparations, the container should be sterile and sealed so as washing with 20 ml of water and drying at 105°, is about 235°,
to exclude micro-organisms. with decomposition (2.4.21).
Labelling. The label states (1) the number of Units per mg; (2) C. Dissolve 50 mg in 2 ml of water, add 1 ml of a 1 per cent w/
whether or not the material is intended for use in the v solution of ninhydrin and heat for a few minutes on a water-
manufacture of parenteral preparations. bath; a violet colour is produced.
Tests
Kanamycin Acid Sulphate
Kanamycin Acid Sulphate is a form of kanamycin sulphate
prepared by adding Sulphuric Acid to a solution of Kanamycin Specific optical rotation (2.4.22). +103° to +115°, determined
Sulphate and subsequent drying. at 20° in a 1.0 per cent w/v solution.
Kanamycin Acid Sulphate has a potency of not less than 650 Kanamycin B. Determine by thin-layer chromatography
Units per mg, calculated on the dried basis. (2.4.17), coating the plate with a 0.75-mm layer of the following
mixture. Mix 0.3 g of carbomer (such as Carbopol 934) with
Description. A white or almost white powder; odourless or 240 ml of water and allow to stand, with moderate shaking, for
almost odourless; hygroscopic. 1 hour; adjust to pH 7.0 by the gradual addition, with
continuous shaking, of 2 M sodium hydroxide and add 30 g
Identification of silica gel H. Heat the plate at 110° for 1 hour, allow to cool
A. Determine by thin-layer chromatography (2.4.17), coating and use immediately.
the plate with a 0.75-mm layer of the following mixture. Mix 0.3 Mobile phase. A 7 per cent w/v solution of potassium
g of carbomer (such as Carbopol 934) with 240 ml of water dihydrogen phosphate.
644
IP 2007 KANAMYCIN INJECTION
Test solution. A 0.5 per cent w/v solution of the substance Kanamycin Injection
Kanamycin Injection is either a sterile solution of Kanamycin
Reference solution. A 0.02 per cent w/v solution of kanamycin Sulphate in Water for Injections containing Sulphuric Acid
B sulphate RS in water. and suitable buffering and stabilising agents or, is a sterile
Apply to the plate 4 µl of each solution. Allow the mobile material consisting of Kanamycin Acid Sulphate with buffering
phase to rise 12 cm. Dry the plate in a current of warm air, agents and other excipients. It is filled in a sealed container.
spray with ninhydrin and stannous chloride reagent and
A. Kanamycin Injection (Solution)
heat at 110° for 15 minutes. Any spot corresponding to
kanamycin B in the chromatogram obtained with the test Kanamycin Injection contains not less than 97.0 per cent and
solution is not more intense than the spot in the chromatogram not more than 110.0 per cent of the stated number of Units of
obtained with the reference solution. kanamycin.
Sulphates. 23.0 to 26.0 per cent of SO4, calculated on the dried Description. A colourless to pale yellow solution.
basis and determined by the following method. Dissolve 0.25
g in 100 ml of water and adjust the pH to 11 using strong Identification
ammonia solution. Add 10.0 ml of 0.1 M barium chloride and Determine by thin-layer chromatography (2.4.17), coating the
0.5 mg of metalphthalein. Titrate with 0.1 M disodium plate with a 0.75-mm layer of the following mixture. Mix 0.3 g of
edetate; when the colour of the solution begins to change, carbomer (such as Carbopol 934) with 240 ml of water and
add 50 ml of ethanol (95 per cent) and continue the titration allow to stand, with moderate shaking, for 1 hour; adjust the
until the blue colour disappears. pH to 7.0 by the gradual addition, with continuous shaking, of
1 ml of 0.1 M barium chloride is equivalent to 0.009606 g of 2 M sodium hydroxide and add 30 g of silica gel H. Heat the
SO4. plate at 110° for 1 hour, allow to cool and use immediately.
Sulphated ash (2.3.18). Not more than 0.5 per cent. Mobile phase. A 7 per cent w/v solution of potassium
on 1.0 g by drying in an oven at 60° at a pressure not exceeding Test solution. A suitable volume diluted with water to contain
0.7 kPa for 3 hours. 800 Units per ml.
kanamycin sulphate RS in water.
Method A or B (2.2.10).
Reference solution (b). A solution containing 0.1 per cent w/
Kanamycin Acid Sulphate intended for use in the
v each of kanamycin sulphate RS, neomycin sulphate RS and
streptomycin sulphate RS in water.
endotoxins complies with the following additional Apply to the plate 10 µl of each solution. Allow the mobile
requirement. phase to rise 12 cm. Dry the plate in a current of warm air,
spray with a mixture of equal volumes of a 0.2 per cent w/v
Bacterial endotoxins (2.2.3). Not more than 0.67 Endotoxin solution of 1,3-napthalenediol in ethanol (95 per cent) and
Unit per mg of kanamycin. a 45 per cent w/v of solution of sulphuric acid and heat at
Kanamycin Sulphate intended for use in the manufacture of 150° for 5 to 10 minutes. The principal spot in the chromatogram
parenteral preparations without a further appropriate obtained with the test solution corresponds to that in the
sterilisation procedure complies with the following chromatogram obtained with reference solution (a). The test
additional requirement. is not valid unless the chromatogram obtained with reference
solution (b) shows three clearly separated spots.
Storage. Store protected from light and moisture. If the material
Tests
is intended for use in the manufacture of parenteral pH (2.4.24). 4.0 to 6.0.
preparations, the container should be sterile and sealed so as
Kanamycin B. Determine by thin-layer chromatography
to exclude micro-organisms.
(2.4.17), coating the plate with a 0.75-mm layer of the following
Labelling. The label states (1) the number of Units per mg; (2) mixture. Mix 0.3 g of carbomer (such asCarbopol 934) with 240
whether or not the material is intended for use in the ml of water and allow to stand, with moderate shaking, for 1
manufacture of parenteral preparations. hour; adjust to pH 7.0 by the gradual addition, with continuous
645
KANAMYCIN INJECTION IP 2007
shaking, of 2 M sodium hydroxide and add 130 g of silica gel 2 M sodium hydroxide and add 30 g of silica gel H. Heat the
H. Heat the plate at 110° for 1 hour, allow to cool and use plate at 110° for 1 hour, allow to cool and use immediately.
immediately.
Mobile phase. A 7 per cent w/v solution of potassium
Mobile phase. A 7 per cent w/v solution of potassium dihydrogen phosphate.
Test solution. A suitable volume diluted with water to contain
Test solution. A suitable volume diluted with water to contain 800 Units per ml.
3750 Units per ml.
Reference solution. A 0.02 per cent w/v solution of kanamycin kanamycin sulphate RS in water.
B sulphate RS in water.
Apply to the plate 4 µl of each solution. Allow the mobile w/v each of kanamycin sulphate RS, neomycin sulphate RS
phase to rise 12 cm. Dry the plate in a current of warm air, and streptomycin sulphate RS in water.
heat at 110° for 15 minutes. Any spot corresponding to Apply to the plate 10 µl of each solution. Allow the mobile
kanamycin B in the chromatogram obtained with the test phase to rise 12 cm. Dry the plate in a current of warm air,
solution is not more intense than the spot in the chromatogram spray with a mixture of equal volumes of a 0.2 per cent w/v
obtained with the reference solution. solution of 1,3-napthalenediol in ethanol (95 per cent) and
a 45 per cent w/v of solution of sulphuric acid and heat at
Bacterial endotoxins (2.2.3). Not more than 0.67 Endotoxin 150° for 5 to 10 minutes. The principal spot in the chromatogram
Unit per mg of kanamycin. obtained with the test solution corresponds to that in the
Assay. Determine by the microbiological assay of antibiotics, chromatogram obtained with reference solution (a). The test
Method A or B (2.2.10). is not valid unless the chromatogram obtained with reference
solution (b) shows three clearly separated spots.
The upper fiducial limit of error is not less than 97.0 per cent
and the lower fiducial limit of error is not more than 110.0 per Tests
cent of the stated number of Units.
B. Kanamycin Injection (Powder)
Kanamycin B. Determine by thin-layer chromatography
The injection is constituted by dissolving the contents of the (2.4.17), coating the plate with a 0.75-mm layer of the following
sealed container in the requisite amount of sterile Water for mixture. Mix 0.3 g of carbomer (such as Carbopol 934) with
Injections, immediately before use. 240 ml of water and allow to stand, with moderate shaking, for
The constituted solution complies with the requirements for 1 hour; adjust to pH 7.0 by the gradual addition, with
Clarity of solution and Particulate matter stated under continuous shaking, of 2 M sodium hydroxide and add 30 g
Parenteral Preparations (Injections). of silica gel H. Heat the plate at 110° for 1 hour, allow to cool
and use immediately.
after preparation but, in any case, within the period Mobile phase. A 7 per cent w/v solution of potassium
recommended by the manufacturer. dihydrogen phosphate.
Kanamycin Injection contains not less than 95.0 per cent and Test solution. A suitable volume diluted with water to contain
not more than 115.0 per cent of the stated number of Units of 3750 Units per ml.
kanamycin Reference solution. A 0.02 per cent w/v solution of kanamycin
The contents of the sealed container comply with the B sulphate RS in water.
requirements stated under Parenteral Preparations Apply to the plate 4 µl of each solution. Allow the mobile
(Powders for Injection) and with the following requirements. phase to rise 12 cm. Dry the plate in a current of warm air,
Identification heat at 110° for 15 minutes. Any spot corresponding to
kanamycin B in the chromatogram obtained with the test
plate with a 0.75-mm layer of the following mixture. Mix 0.3 g of
carbomer (such as Carbopol 934) with 240 ml of water and
allow to stand, with moderate shaking, for 1 hour; adjust the Bacterial endotoxins (2.2.3). Not more than 0.67 Endotoxin
pH to 7.0 by the gradual addition, with continuous shaking, of Unit per mg of kanamycin.
646
IP 2007 LIGHT KAOLIN
Assay. Determine the weight of the contents of 10 containers. water, 10 ml of hydrochloric acid and 25 ml of 4-methylpentan-
Using the mixed contents of the 10 containers determine by 2-one, shake for 2 minutes, allow the layers to separate and
the microbiological assay of antibiotics, Method A or Method evaporate the aqueous layer to dryness on a water-bath.
B (2.2.10). Dissolve the residue in 1 ml of 5 M acetic acid, dilute to 25 ml
For a container of average content weight, the upper fiducial with water and filter. 12 ml of the resulting solution complies
limit of error is not less than 95.0 per cent and the lower fiducial with the limit test for heavy metals, Method D (50 ppm). Use
limit of error is not more than 115.0 per cent of the stated lead standard solution (1 ppm Pb) to prepare the standard.
number of Units. Chlorides (2.3.12). Shake 4.0 g with a mixture of 34 ml of
distilled water and 6 ml of 5 M acetic acid for 1 minute and
Storage. Store in single dose containers protected from light
filter. 10 ml of the filtrate complies with the limit test for chlorides
and moisture.
(250 ppm).
Labelling. The label states (1) the strength in terms of the Sulphates (2.3.17). 2 ml of the filtrate obtained in the test for
number of Units or the equivalent weight of kanamycin in a chlorides diluted to 15 ml complies with the limit test for
suitable dose-volume or in the sealed container; (2) the volume sulphates (750 ppm).
of Water for Injections for constituting the solution (for
contents of a sealed container). Substances soluble in mineral acids. Not more than 1 per
cent, determined by the following method. To 10 ml of solution
A add 1.5 ml of 1 M sulphuric acid, evaporate to dryness on
a water-bath, ignite, cool and weigh.
Heavy Kaolin Organic impurities. Heat 0.3 g to redness in a calcination
Heavy Kaolin is a purified, natural, hydrated aluminium silicate tube. The residue is only slightly more coloured than the
of variable composition. original substance.
Description. Fine, white or greyish white, soft powder; Adsorption power. In a ground-glass-stoppered test-tube
odourless. shake 1.0 g with 10 ml of a 0.37 per cent w/v solution of
methylene blue for 2 minutes and allow to settle. Centrifuge
Identification and dilute 1 volume of the solution to 100 volumes with water.
The solution is not more intensely coloured than a 0.003 per
A. To 0.5 g in a metal crucible add 1 g of potassium nitrate and cent w/v solution of methylene blue.
3 g of anhydrous sodium carbonate, heat until the mixture
has melted and allow to cool. To the residue add 20 ml of Swelling power. Triturate 2 g with 2 ml of water; the mixture
boiling water, mix, filter and wash the residue with 50 ml of does not flow.
water. To the residue add 1 ml of hydrochloric acid and 5 ml Loss on ignition (2.4.20). Not more than 15.0 per cent,
of water and filter. To the filtrate add 1 ml of 10 M sodium determined on 1.0 g.
hydroxide and filter. To the filtrate add 3 ml of ammonium
chloride solution; a gelatinous, white precipitate is produced.
B. 0.25 g gives the reaction of silicates (2.3.1). Light Kaolin
Light Kaolin is a native hydrated aluminium silicate, freed
Tests
from most of its impurities by elutriation and dried. It may
Acidity or alkalinity. To 1.0 g add 20 ml of carbon dioxide- contain a suitable dispersing agent.
free water, shake for 2 minutes and filter. To 10 ml of the filtrate Description. Light, white powder free from gritty particles;
add 0.1 ml of phenolphthalein solution. The solution is odourless; almost tasteless; unctuous to the touch.
colourless and not more than 0.25 ml of 0.01 M sodium
hydroxide is required to change the colour of the solution to Identification
pink.
A. Fuse 1 g with 2 g of anhydrous sodium carbonate, warm
Arsenic (2.3.10). Disperse 5.0 g in 50 ml of water and add 10 ml the residue with 10 ml of water, filter, wash the filter with 5 ml
of stannated hydrochloric acid. The resulting solution of water and reserve the residue. To the combined filtrate and
complies with the limit test for arsenic (2 ppm). washings add 3 ml of hydrochloric acid; a gelatinous
Heavy metals (2.3.13). Boil 5.0 g with 7.5 ml of 2 M hydrochloric precipitate is produced.
acid and 27.5 ml of water for 5 minutes, filter, wash the residue B. Dissolve the residue reserved in test A in 10 ml of 2 M
with water and dilute the combined filtrate and washings to hydrochloric acid; the solution gives reaction B of aluminium
50 ml with water (solution A). To 5 ml of solution A add 5 ml of salts (2.3.1).
647
KETAMINE HYDROCHLORIDE IP 2007
C. Triturate 2 g with 2 ml of water; the resulting mixture flows. Loss on ignition (2.4.20). Not more than 15.0 per cent,
Coarse particles. Transfer 5 g to a stoppered cylinder (16 cm × determined on 1 g by igniting at 600°.
35 mm), add 60 ml of a 1 per cent w/v solution of sodium Storage . Store protected from moisture.
pyrophosphate, shake thoroughly and allow to stand for
5 minutes. Using a pipette, withdraw 50 ml from a point about
5 cm below the surface of the liquid. To the remaining liquid
add 50 ml of water, shake, allow to stand for 5 minutes and Ketamine Hydrochloride
withdraw 50 ml in the same manner as before. Repeat the
operation until a total of 400 ml of suspension has been Cl
O
withdrawn under the prescribed conditions. Transfer the
, HCl
remainder to an evaporating dish and evaporate to dryness
on a water-bath. The residue, after drying at 105°, weighs not NHCH3
more than 25 mg.
Fine particles. Disperse 5 g in 250 ml of water by shaking C13H16ClNO,HCl Mol. Wt. 274.2
vigorously for 2 minutes in a stoppered flask, pour immediately Ketamine Hydrochloride is (RS)-2-(2-chlorophenyl)-2-
into a glass cylinder, 5 cm in diameter, and transfer 20 ml to a methylaminocyclohexanone hydrochloride.
glass dish using a pipette. Evaporate to dryness and dry to
Ketamine Hydrochloride contains not less than 98.5 per cent
constant weight at 105°. Allow the remainder of the suspension
and not more than 101.0 per cent of C13H16ClNO,HCl.
to stand for 4 hours at 20° and withdraw a second 20-ml portion
using a pipette with its tip exactly 5 cm below the surface and Description. A white, crystalline powder.
without disturbing the sediment. Transfer the second portion
to a glass dish, evaporate to dryness and dry to constant Identification
weight at 105°. The weight of the residue from the second
portion is not less than 70 per cent of the weight of the residue
Compare the spectrum with that obtained with ketamine
from the first portion.
hydrochloride RS or with the reference spectrum of ketamine
Tests hydrochloride.
B. A 10 per cent w/v solution gives the reactions of chlorides
Arsenic (2.3.10). Disperse 5 g in 50 ml of water and add 10 ml
(2.3.1).
of stannated hydrochloric acid; the resulting solution
complies with the limit test for arsenic (2 ppm). C. Melting range (2.4.21). 258° to 261°.
Heavy metals. Heat 6.0 g for 15 minutes under a reflux Tests
condenser on a water-bath with a mixture of 70 ml of water
and 10 ml of hydrochloric acid and filter. To 40 ml of the Appearance of solution. A 20.0 per cent w/v solution is clear
filtrate add 0.5 ml of nitric acid and evaporate to a low bulk. (2.4.1), and colourless (2.4.1).
Add 20 ml of water, 2 g of ammonium chloride and 2 g of pH (2.4.24). 3.5 to 4.1, determined in a 10.0 per cent w/v solution.
ammonium thiocyanate and extract with two quantities, each
of 10 ml, of a mixture of equal volumes of amyl alcohol and Foreign amines. Determine by thin-layer chromatography
ether. To the aqueous layer add 2 g of citric acid and sufficient (2.4.17), coating the plate with silica gel.
water to produce 60 ml. 12 ml of the solution complies with the Mobile phase. A mixture of 80 volumes of toluene, 19.5 volumes
limit test for heavy metals, Method D (2.3.13) (20 ppm). of 2-propanol and 0.5 volume of strong ammonia solution.
Chlorides (2.3.12). Boil 1.0 g with 40 ml of water and 20 ml of Test solution. Dissolve 0.6 g of the substance under
2 M nitric acid under a reflux condenser for 5 minutes, cool examination in 10 of methanol.
and filter. 30 ml of the filtrate complies with the limit test for
chlorides (330 ppm).
Soluble matter. Boil 2 g with 100 ml of 0.2 M hydrochloric
acid under a reflux condenser for 5 minutes, cool, filter and
dry the plate in air and spray with freshly prepared acid
evaporate 50 ml to dryness. The residue, after ignition at about
potassium iodobismuthate solution. Any secondary spot in
600o for 30 minutes, weighs not more than 10 mg.
Loss on drying (2.4.19). Not more than 1.5 per cent, determined intense than the spot in the chromatogram obtained with the
on 1 g by drying in an oven at 105°. reference solution.
648
IP 2007 KETOCONAZOLE
Related substances. Determine by thin-layer chromatography a solution containing 0.07 per cent w/v of ketamine. When
(2.4.17), coating the plate with silica gel. examined in the range 230 nm to 360 nm (2.4.7), the resulting
Mobile phase. A mixture of 80 volumes of toluene, 19.5 volumes solution shows an absorption maximum at about 301 nm.
of 2-propanol and 0.5 volume of strong ammonia solution. Tests
pH (2.4.24). 3.0 to 5.5.
Unit per mg of ketamine hydrochloride.
Assay. To an accurately measured volume containing about
0.5 g of ketamine add sufficient water to produce 200.0 ml and
dry the plate in air and expose to iodine vapour for about an
mix. To 20.0 ml of the resulting solution add 3 ml of 0.1 M
hour. Any secondary spot in the chromatogram obtained with
sodium hydroxide and extract with three quantities, each of
15 ml, of chloroform. Combine the chloroform extracts and
shake successively with three quantities, each of 30 ml, of
0.05 M sulphuric acid. Dilute the combined acid extracts to
200.0 ml with 0.05 M sulphuric acid (saturated with
Heavy metals (2.3.13). 1.0 g complies with limit test for heavy chloroform), and measure the absorbance of the resulting
metals, Method A (20 ppm). solution at the maximum at about 269 nm (2.4.7).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Calculate the content of C13H16ClNO from the absorbance
Assay. Weigh accurately about 0.5 g and dissolve in 1 ml of obtained by repeating the determination using a standard
anhydrous formic acid. Add 50 ml of anhydrous glacial acetic solution of ketamine hydrochloride RS containing the
acid and 10 ml of mercuric acetate solution. Titrate with 0.1 equivalent of 250 µg per ml of ketamine base in the same
M perchloric acid, using crystal violet solution as indicator. medium in place of the substance under examination.
Carry out a blank titration. Storage. Store protected from light at a temperature not
1 ml of 0.1 M perchloric acid is equivalent to 0.02742 g of exceeding 30°.
C13H16ClNO,HCl. Labelling. The label states the strength in terms of the
Storage. Store protected from moisture. equivalent amount of ketamine in a suitable dose-volume.
Ketoconazole
Ketamine Injection
Ketamine Hydrochloride Injection N
Ketamine Injection is a sterile solution of Ketamine N

Hydrochloride in Water for Injections. O
O
Ketamine Injection contains not less than 95.0 per cent and N N O
not more than 105.0 per cent of the stated amount of ketamine, O
H 3C H Cl
C13H16ClNO. Cl
Identification C26H28Cl2N4O4 Mol. Wt. 531.4
A. Dilute a suitable volume with 0.1 M hydrochloric acid to Ketoconazole is cis-1-acetyl-4-[[(2RS,4RS)-2-(2,4-

produce a solution containing 0.03 per cent w/v of ketamine. dichlorophenyl)-2-(1H-imidazol-1-ylmethyl)-1,3-dioxolan-4-
When examined in the range 230 nm to 360 nm (2.4.7), the yl]methoxyl]phenylpiperazine.
resulting solution shows absorption maxima at about 269 nm Ketoconazole contains not less than 98.0 per cent and not
and 276 nm. more than 102.0 per cent of C26H28Cl2N4O4, calculated on the
B. Dilute a suitable volume with a mixture of 49 volumes of dried basis.
methanol and 1 volume of 1 M sodium hydroxide to produce Description. A white to off-white, crystalline powder.
649
KETOCONAZOLE TABLETS IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). Determine by thin-layer chromatography (2.4.17), coating the
Compare the spectrum with that obtained with ketoconazole plate with silica gel G.
RS or with the reference spectrum of ketoconazole.
Mobile phase. A mixture of 42 volumes of hexane, 40 volumes
B. In the test for Related substances, the principal spot in the of ethyl acetate, 15 volumes of methanol, 2 volumes of water
chromatogram obtained with the test solution corresponds to and 1 volume of glacial acetic acid.
C. Melting range (2.4.21). 148° to 152°. containing 200 mg of Ketoconazole with 10 ml of chloroform,
dilute to 20 ml with chloroform and filter.
Tests
Specific optical rotation (2.4.22). –1.0° to +1.0°, determined in ketoconazole RS in chloroform.
a 4.0 per cent w/v solution in methanol.
ketoconazole RS in chloroform.
Mobile phase. A mixture of 42 volumes of hexane, 40 volumes Apply to the plate 10 µl of each solution. After development,
of ethyl acetate, 15 volumes of methanol, 2 volumes of water dry the plate in air and expose to iodine vapour. Any secondary
and 1 volume of glacial acetic acid. spot in the chromatogram obtained with the test solution is
not more intense than the spot in the chromatogram obtained
Test solution. Dissolve 0.1 g of the substance under with reference solution (b).
ketoconazole RS in chloroform. Assay. Determine by liquid chromatography (2.4.14).
Reference solution (b). A 0.02 per cent w/v solution of Test solution. Weigh and powder 20 tablets. Weigh accurately
ketoconazole RS in chloroform. a quantity of the powder containing about 200 mg of
Apply to the plate 10 µl of each solution. After development, Ketoconazole, shake with 50.0 ml of a mixture of equal volumes
dry the plate in air and expose to iodine vapour. Any secondary of methanol and dichloromethane and centrifuge. To 5.0 ml
spot in the chromatogram obtained with the test solution is of this solution add 5.0 ml of a 0.5 per cent w/v solution of
not more intense than the spot in the chromatogram obtained terconazole RS (internal standard) in the methanol-
with reference solution (b). dichloromethane solvent mixture and dilute to 50.0 ml with the
Heavy metals (2.3.13). 1.0 g complies with limit test for heavy same solvent mixture.
metals, Method B (20 ppm). Reference solution. Dissolve 20 mg of ketoconazole RS in
Sulphated ash (2.3.18). Not more than 0.1 per cent. 20 ml of the methanol-dichloromethane solvent mixture, add
5.0 ml of the internal standard solution and dilute to 50.0 ml
2.7 kPa for 4 hours. Chromatographic system
Assay. Weigh accurately about 0.2 g, dissolve in 40 ml of – a stainless steel column 30 cm x 3.9 mm, packed with
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric octadecylsilyl silica gel,
acid, determining the end-point potentiometrically – mobile phase: a mixture of 7 volumes of 0.2 per cent
(2.4.25).Carry out a blank titration. w/v of di-isopropylamine in methanol and 3 volumes
1 ml of 0.1 M perchloric acid is equivalent to 0.02657 g of of a 0.5 per cent w/v solution of ammonium acetate,
C26H28Cl2N4O4. – flow rate. 3 ml per minute,
Ketoconazole Tablets Inject the test solution and the reference solution. The relative
retention times are about 0.6 for ketoconazole and 1.0 for
Ketoconazole Tablets contain not less than 90.0 per cent and
terconazole.
ketoconazole, C26H28Cl2N4O4. Calculate the content of C26H28Cl2N4O4 in the tablets.
650
IP 2007 KETOPROFEN CAPSULES
Ketoprofen Mobile phase. A mixture of 70 volumes of toluene, 30 volumes

of di-isopropyl ether and 1 volume of formic acid.
O CH3 Test solution. A 10 per cent w/v solution of the substance

under examination in acetone.
COOH Reference solution (a). A 0.05 per cent w/v solution of the
C16H14O3 Mol. Wt. 254.3 substance under examination in acetone.
Ketoprofen is (RS)-2-(3-benzoylphenyl)propionic acid. Apply rapidly to the plate 5 µl of each solution. Develop the
chromatogram within 10 minutes of applying the first spot.
Ketoprofen contains not less than 98.5 per cent and not more
After development, dry the plate air and examine in ultraviolet
than 100.5 per cent of C16H14O3, calculated on the dried basis.
Description. A white or almost white, crystalline powder; obtained with the test solution is not more intense than the
odourless or almost odourless. spot in the chromatogram obtained with reference solution (a)
and not more than three such spots are more intense than the
Identification spot in the chromatogram obtained with reference solution
Test A may be omitted if tests B and C are carried out. Tests B (b).
and C may be omitted if test A is carried out. Sulphated ash (2.3.18). Not more than 0.1 per cent.
A. Determine by infrared absorption spectrophotometry (2.4.6). Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Compare the spectrum with that obtained with ketoprofen RS on 1.0 g by drying in an oven at 60° at a pressure not exceeding
or with the reference spectrum of ketoprofen. 0.7 kPa.
B. When examined in the range 230 nm to 360 nm (2.4.7), a Assay. Weigh accurately about 0.5 g, dissolve in 25 ml of
0.001 per cent w/v solution in methanol (75 per cent) shows ethanol (95 per cent) previously neutralised to
an absorption maximum only at about 258 nm; absorbance at phenolphthalein solution, add 25 ml of water Titrate with 0.1
about 258 nm, about 0.66. M sodium hydroxide using phenolphthalein solution as
C. Determine by thin-layer chromatography (2.4.17), coating indicator.
the plate with silica gel GF254. 1 ml of 0.1 M sodium hydroxide is equivalent to 0.02543 g of
Mobile phase. A mixture of 50 volumes of acetone, 49 volumes C16H14O3.
of dichloromethane and 1 volume of glacial acetic acid. Storage. Store protected from moisture.
ketoprofen RS in acetone. Ketoprofen Capsules
Reference solution (b). A mixture of equal volumes of 1.0 per Ketoprofen Capsules contain not less than 92.5 per cent and
cent w/v solution of indomethacin RS and refererence solution not more than 107.5 per cent of the stated amount of
(a) in acetone. ketoprofen, C16H14O3.
Identification
The principal spot in the chromatogram obtained with the test A.Shake a quantity of the contents of the capsules containing
solution corresponds to that in the chromatogram obtained 0.5 g of Ketoprofen with 50 ml of chloroform for 5 minutes,
with reference solution (a). The test is not valid unless the filter, evaporate to dryness using a rotary evaporator, induce
chromatogram obtained with reference solution (b) shows two crystallisation by prolonged scratching of the inside wall of
clearly separated spots. the container with a glass rod and separate the crystals by
centrifugation or filtration.
Tests
Related substances. Determine by thin-layer chromatography Compare the spectrum with that obtained with ketoprofen RS
(2.4.17), coating the plate with silica gel GF254. or with the reference spectrum of ketoprofen.
651
KETOPROFEN CAPSULES IP 2007
B. When examined in the range 230 nm to 360 nm (2.4.7), the After development, dry the plate in air and examine in
final solution obtained in the Assay shows an absorption ultraviolet light at 254 nm. Any secondary spot in the
maximum only at about 258 nm. chromatogram obtained with the test solution is not more
Tests reference solution (a) and not more than three such spots are
of di-isopropyl ether and 1 volume of formic acid. Assay. Weigh accurately a quantity of the mixed contents of
20 capsules containing about 25 mg of Ketoprofen, shake for
Test solution. Shake a quantity of the contents of the capsules
10 minutes with 150 ml of methanol (75 per cent), mix and
containing 0.3 g of Ketoprofen with 10 ml of chloroform, filter,
dilute to 250.0 ml with methanol (75 per cent). Allow to stand,
wash with 5 ml of chloroform, evaporate the combined filtrate
dilute 10.0 ml of the supernatant liquid to 100.0 ml with
and washings to dryness and dissolve the residue in 3 ml of
methanol (75 per cent) and measure the absorbance of the
acetone.
Reference solution (a). Dilute 1 volume of test solution to 100
Calculate the content of C16H14O3 taking 662 as the specific
Reference solution (b). Dilute 1 volume of test solution to 200
Apply rapidly to the plate 5 µl of each solution. Develop the
chromatogram within 10 minutes of applying the first spot.
652
L
Labetalol Hydrochloride ....
Labetalol Tablets ....
Lactic Acid ....
Lactose ....
Lamivudine ....
Lamivudine Oral Solution ....
Lamivudine Tablets ....
Lamivudine And Tenofovir Tablets ....
Lamivudine And Zidovudine Tablets ....
Lamivudine, Nevirapine And Stavudine Tablets ....
Lamivudine, Nevirapine and Stavudine Dispersible Tablets ....
Lamotrigine ....
Lamotrigine Dispersible Tablets ....
Lanatoside C ....
Lanatoside C Tablets ....
Levamisole Hydrochloride ....
Levamisole Tablets ....
Levocetirzine Dihydrochloride ....
Levocetirzine Tablets ....
Levodopa ....
Levodopa And Carbidopa Tablets ....
Levodopa Capsules ....
Levodopa Tablets ....
Levofloxacin Hemihydrate ....
Levofoxacin Infusion ....
Levofloxacin Tablets ....
Levonorgestrel ....
Levonorgestrel And Ethinyloestradiol Tablets ....
Lignocaine Hydrochloride ....
Lignocaine And Adrenaline Injection ....
653
Lignocaine And Dextrose Injection ....

Lignocaine Gel ....
Lignocaine Injection ....
Lincomycin Hydrochloride ....
Lincomycin Capsules ....
Lindane ....
Absorbent Lint ....
Lisinopril ....
Lisinopril Tablets ....
Lithium Carbonate ....
Lithium Carbonate Tablets ....
Lomustine ....
Lomustine Capsules ....
Loperamide Hydrochloride ....
Loperamide Capsules ....
Loperamide Tablets ....
Lopinavir ....
Lopinavir and Ritonavir Capsules ....
Lopinavir And Ritonavir Tablets ....
Losartan Potassium ....
Losartan Tablets ....
Lynoestrenol ....
654
IP 2007 LABETALOL HYDROCHLORIDE
– temperature:
Labetalol Hydrochloride column. 270°,
inlet port and detector at 300°,
OH – flow rate. 20 ml per minute of the carrier gas.
CH3
, HCl Two peaks due to the two diasterioisomers appear in the
N CONH2 chromatogram. Adjust the sensitivity of the detector so that
H OH in the chromatogram obtained, the height of the taller of the
diasterioisomer peaks is about 80 per cent of the full-scale
deflection. The area of each peak is not less than 45 per cent
C19H24N2O3,HCl Mol. Wt. 364.9
and not more than 55 per cent of the sum of the areas of the
Labetalol Hydrochloride is all-rac-2-hydroxy-5-[1-hydroxy- two peaks. The test is not valid unless the height of the trough
2-(1-methyl-3-phenylpropylamino)ethyl]benzamide separating the two diasterioisomers peaks is less than 5 per
hydrochloride. cent of the full-scale deflection.
Labetalol Hydrochloride contains not less than 98.5 per cent Related substances. Determine by liquid chromatography
and not more than 101.0 per cent of C19H24N2O3,HCl, calculated (2.4.14).
on the dried basis.
Description. A white or almost white powder or granules. examination in the mobile phase and dilute to 10 ml with the
mobile phase.
Identification
Reference solution. Dilute 0.5 ml of the test solution to 100 ml
A. Determine by infrared absorption spectrophotometry (2.4.6). with the mobile phase.
Compare the spectrum with that obtained with labetalol
hydrochloride RS or with the reference spectrum of labetalol Chromatographic system
hydrochloride. – a stainless steel column 15 cm ´ 4.6 mm, packed with
octadecylsilane chemically bonded to porous silica
B. When examined in the range 230 nm to 360 nm (2.4.7), a (5 µm),
0.005 per cent w/v solution in 0.1 M hydrochloric acid shows – column. temperature
an absorption maximum only at 302 nm; absorbance at 302 nm, – mobile phase: a degassed mixture of 150 ml of
about 0.43. tetrahydrofuran, 300 ml of methanol, 550 ml of water,
C. Dissolve 10 mg in 5 ml of water and add 1 ml of ferric 0.82 g of tetrabutylammonium hydrogen sulphate, 1 g
chloride test solution; a purple colour is produced. of sodium octyl sulphate and 10 ml of a 10 per cent w/v
solution of sulphuric acid,
D. A 1 per cent w/v solution gives the reactions of chlorides
(2.3.1).
Tests – a 20 µl loop injector.
Appearance of solution. A 1.0 per cent w/v solution is clear Equilibrate the column with the mobile phase for about
(2.4.1), and not more intensely coloured than reference solution 30 minutes.
YS6 (2.4.1). Adjust the sensitivity of the system so that the height of the
pH (2.4.24). 4.0 to 5.0, determined in a 1.0 per cent w/v solution. principal peak in the chromatogram obtained with the reference
solution is at least 50 per cent of the full scale of the recorder.
Diasterioisomer ratio. Determine by gas chromatography The retention time of the principal peak is 10 minutes to
(2.4.13). 15 minutes. If necessary, adjust the water content of the mobile
Test solution. Dissolve 2 mg of the substance under phase ensuring that the 2:1 ratio of methanol to
examination in 1 ml of a 1.2 per cent w/v solution of tetrahydrofuran is maintained.
1-butaneboronic acid in anhydrous pyridine and allow to Inject each solution. Continue the chromatography for three
stand for 20 minutes. times the retention time of the principal peak in the
Chromatographic system chromatogram obtained with the test solution; the area of any
– a stainless steel column 1.5 m x 4 mm, packed with acid- peak other than the principal peak is not greater than 0.6 times
washed, silanised diatomaceous support (125 to 150 that of the principal peak in the chromatogram obtained with
µm) coated with 3 per cent w/w of polymethyl phenyl the reference solution (0.3 per cent); the sum of the areas of
siloxane, any such peaks is not greater than the area of the principal
655
LABETALOL TABLETS IP 2007
peak in the chromatogram obtained with the reference solution C. Disperse a quantity of the powered tablets containing
(0.5 per cent). Ignore any peak due to the solvent and any 10 mg of Labetalol Hydrochloride in a mixture of 2 ml of water
peak with an area less than 0.1 times the area of the principal and 2 ml of ferric chloride test solution; a purple colour is
peak in the chromatogram obtained with the reference solution. produced.
Heavy metals (2.3.13). Dissolve 2.0 g in a mixture of 20 ml of
Tests
water and 5 ml of dilute sodium hydroxide solution. The
solution complies with the limit test for heavy metals, Method Diasterioisomer ratio. Determine by gas chromatography
C (20 ppm). (2.4.13).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Test solution. Shake a quantity of the powdered tablets
containing 0.5 g of Labetalol Hydrochloride with 10 ml of
methanol, filter and evaporate the filtrate to dryness using a
rotary evaporator.
0.7 kPa.
– a stainless steel column 1.5 m ´ 4 mm, packed with acid-
anhydrous formic acid and 40 ml of acetic anhydride. Titrate
washed, silanised diatomaceous support (125 to 150
with 0.1 M perchloric acid, determining the end-point
mm) coated with 3 per cent w/w of polymethyl phenyl
siloxane,
1 ml of 0.1 M perchloric acid is equivalent to 0.03649 g of – temperature:
C19H24N2O3,HCl. column. 270°,
inlet port and detector at 300°,
NOTE - Mix thoroughly throughout the titration and stop
the titration immediately after the end-point is reached.
Two peaks due to the two diasterioisomers appear in the
chromatogram. Adjust the sensitivity of the detector so that
in the chromatogram obtained, the height of the taller of the
diasterioisomer peaks is about 80 per cent of the full-scale
deflection. The area of each peak is not less than 45 per cent
Labetalol Tablets and not more than 55 per cent of the sum of the areas of the
two peaks. The test is not valid unless the height of the trough
Labetalol HydrochlorideTablets separating the two diasterioisomers peaks is less than 5 per
Labetalol Tablets contain not less than 95.0 per cent and not cent of the full-scale deflection.
more than 105.0 per cent of the stated amount of labetalol Related substances. Determine by thin-layer chromatography
hydrochloride, C19H24N2O3,HCl. (2.4.17), coating the plate with silica gel GF254.
Identification Mobile phase. A mixture of 75 volumes of dichloromethane,

A. To a quantity of the powdered tablets containing 50 mg of solution.
Labetalol Hydrochloride add 50 ml of 0.1 M hydrochloric
Test solution. Shake a quantity of the powered tablets
acid and heat on a water-bath for 30 minutes. Cool, filter, add
containing 0.5 g of Labetalol Hydrochloride with 10 ml of
10 ml of ammonia buffer pH 10.0 and extract with three
methanol, filter and use the filtrate.
quantities, each of 15 ml, of dichloromethane. Shake the
combined extracts with 5 g of anhydrous sodium sulphate, Reference solution (a). Dilute 1 volume of the test solution to
filter and evaporate the filtrate to dryness. 100 volumes with methanol.
On the residue, determine by infrared absorption Reference solution (b). Dilute reference solution (a) with an
spectrophotometry (2.4.6). Compare the spectrum with that equal volume of methanol.
obtained with labetalol RS or with the reference spectrum of
labetalol.
dry the plate in a current of warm air, heat at 105° for 30 minutes,
B. When examined in the range 230 nm to 360 nm (2.4.7), a cool and examine in ultraviolet light at 254 nm. Any secondary
0.004 per cent w/v solution of the residue obtained in test A in spot in the chromatogram obtained with the test solution is
0.1 M sodium hydroxide shows an absorption maximum only not more intense than the spot in the chromatogram obtained
at about 333 nm. with reference solution (a) and not more than one such spot is
656
IP 2007 LACTIC ACID
more intense than the spot in the chromatogram obtained Heavy metals (2.3.13). Dissolve 5.0 g in 42 ml of 1 M sodium
with reference solution (b). hydroxide and dilute to 50 ml with distilled water. 12 ml of the
Other tests. Comply with the tests stated under Tablets. resulting solution complies with the limit test for heavy metals,
Method D (10 ppm).
quantity of the powder containing about 0.5 g of Labetalol Citric, oxalic and phosphoric acids. To 5 ml of the solution
Hydrochloride, shake with 200 ml of 0.05 M sulphuric acid prepared in the test for Heavy metals add 6 M ammonia until
for 30 minutes and dilute to 250.0 ml with 0.05 M sulphuric slightly alkaline. Add 1 ml of calcium chloride solution and
acid, mix and filter. Dilute 10.0 ml of the filtrate to 250.0 ml with heat on a water-bath for 5 minutes. Both before and after
the same solvent and measure the absorbance of the resulting heating, any opalescence in the solution is not more intense
solution at the maximum at about 302 nm (2.4.7). than that in a mixture of 5 ml of the test solution and 1 ml of
water.
Calculate the content of C19H24N2O3, HCl taking 86 as the
specific absorbance at 302 nm. Ether-insoluble substances. Dissolve 1.0 g in 25 ml of ether;
the solution is not more opalescent than the solvent used for
the test.
Volatile fatty acids. Cautiously heat 5 g in a glass-stoppered
flask at 50° for 10 minutes; no unpleasant odour resembling
that of the lower fatty acids is recognisable immediately after
Lactic Acid opening the flask.
H OH Methanol and methyl esters. Place 2.0 g in a ground-glass-

stoppered round-bottom flask and add 10 ml of water. Cool in
H3C COOH ice, cautiously add 30 ml of a 30 per cent w/v solution of
potassium hydroxide and cool in ice for a further 10 to
C3H6O3 Mol. Wt. 90.1 15 minutes. Steam distil the mixture into a 10-ml graduated
Lactic Acid consists of a mixture of 2-hydroxypropionic acid, cylinder containing 1 ml of ethanol, collecting a volume of at
its condensation products, such as lactoyl-lactic acid and least 9.5 ml and dilute to 10.0 ml with water. To 1.0 ml of the
other polylactic acids, and water. The equilibrium between distillate add 5 ml of potassium permanganate and phosphoric
lactic acid and polylactic acids depends on concentration and acid solution and mix. After 15 minutes add 2 ml of oxalic
temperature. acid and sulphuric acid solution, stir with a glass rod until
It is usually in the form of the racemate [(RS)-lactic acid], but the solution is colourless and then add 5 ml of decolorised
in some cases the (S)-isomer may predominate. magenta solution. After 2 hours any colour in the solution is
not more intense than that of 1 ml of a reference solution
Lactic Acid contains the equivalent of not less than 88.0 per containing 100 µg of methanol and 0.1 ml of ethanol treated in
cent and not more than 92.0 per cent w/w of C3H6O3. the same manner beginning at the words “add 5 ml of
Description. A colourless or slightly yellow, viscous liquid; potassium permanganate and phosphoric acid solution..”.
almost odourless; hygroscopic. Reducing sugars. Dilute 1 g with 10 ml of water, neutralise
with sodium hydroxide solution, add 5 ml of potassium cupri-
Identification tartrate solution, and boil; no red or greenish precipitate is
A. Warm 1 g with 0.1 g of potassium permanganate; produced.
acetaldehyde is evolved. Sulphated ash (2.3.18). Not more than 0.1 per cent.
B. Gives reaction A of lactates (2.3.1). Assay. Weigh accurately about 1.0 g in a ground-glass-
C. A 10 per cent w/v solution is strongly acidic. stoppered flask and add 10 ml of water. Add 20.0 ml of 1 M
sodium hydroxide, stopper the flask and allow to stand for
Tests 30 minutes. Titrate the excess of alkali with 1 M hydrochloric
acid, using dilute phenolphthalein solution as indicator until
Appearance of solution. The substance under examination is the pink colour is discharged.
not more intensely coloured than reference solution YS6 (2.4.1).
1 ml of 1 M sodium hydroxide is equivalent to 0.09008 g of
C3H6O3.
stannated hydrochloric acid. The resulting solution complies
with the limit test for arsenic (1 ppm). Storage. Store protected from light.
657
LACTOSE IP 2007
Lactose Light absorption (2.4.7). Dissolve 1.0 g in boiling water and

dilute to 10 ml with the same solvent (solution A). Absorbance
Lactose Monohydrate; Milk Sugar of solution A measured at the maximum at about 400 nm, not
more than 0.04.
HO Dilute 1 ml of solution A to 10 ml with water.
O When examined in the range 210 nm to 300 nm, absorbance is
HO not more than 0.25 in the range 210 nm to 220 nm and not more
OH
than 0.07 in the range 270 nm to 300 nm.
HO O O OH , H2O
OH Arsenic (2.3.10). Dissolve 10.0 g in 50 ml of water and add
OH 10 ml of stannated hydrochloric acid AsT; the resulting
OH Heavy metals (2.3.13). Dissolve 4.0 g in 20 ml of warm water,
1.0 ml of 0.1 M hydrochloric acid and sufficient water to
C12H22O11,H2O Mol. Wt. 360.3
produce 25 ml. The solution complies with the limit test for
Lactose is O-β-D-galactopyranosyl-(1 → 4)-α -D- heavy metals, Method A (5 ppm).
glucopyranose monohydrate.
Microbial contamination (2.2.9). Total microbial count not more
Description. A white or almost white, crystalline powder; than 100 per g; 1 g is free from Escherichia coli and salmonellae.
odourless.
Sulphated ash. Not more than 0.1 per cent, determined in the
Identification following manner. To 1.0 g add 1 ml of sulphuric acid, evaporate
to dryness on a water-bath and ignite to constant weight.
and C may be omitted if test A is carried out. Water (2.3.43). 4.5 to 5.5 per cent, determined on 0.5 g.
A. Determine by infrared absorption spectrophotometry (2.4.6). Storage. Store protected from moisture.
Compare the spectrum with that obtained with lactose RS or
with the reference spectrum of lactose.
B. To 5 ml of a saturated solution add 5 ml of 1 M sodium
hydroxide and gently warm the mixture; the liquid becomes
Lamivudine
yellow and then brownish-red. Cool to room temperature and
add 0.2 ml of potassium cupri-tartrate solution; a red NH2
precipitate is formed.
N
C. Heat 5 ml of a 5 per cent w/v solution with 5 ml of 10 M
ammonia in a water-bath at 80° for 10 minutes; a red colour O N
develops. O
Tests HO
S
Appearance of solution. Dissolve 1.0 g in water by heating to
50°, dilute to 10 ml with water and allow to cool. The solution C8H11N3O3S Mol. Wt. 229.3
is clear (2.4.1), and not more intensely coloured than reference Lamivudine is (2R,5S)-4-amino-1-[2-(hydroxymethyl)-1,3-
solution BYS7 (2.4.1). oxathiolan-5yl]-2(1H)-pyrimidinone.
Acidity or alkalinity. Dissolve 6 g in 25 ml of carbon dioxide- Lamivudine contains not less than 98.0 per cent and not more
free water by boiling, cool and add 0.3 ml of phenolphthalein than 102.0 per cent of C8H11N3O3S, calculated on the anhydrous
solution. The solution is colourless and not more than 0.4 ml basis.
of 0.1 M sodium hydroxide is required to change the colour of
the solution to pink. Description. A white or almost white powder.
in a solution obtained by dissolving 10.0 g in 80 ml of water
by heating to 50°, allowing to cool, adding 0.2 ml of 6 M A. Determine by infrared absorption spectrophotometry (2.4.6).
ammonia, allowing to stand for 30 minutes and diluting to Compare the spectrum with that obtained with lamivudine RS
100.0 ml with water. . or with the reference spectrum of lamivudine.
658
IP 2007 LAMIVUDINE TABLETS
B. In the Assay, the principal peak in the chromatogram – mobile phase: a degassed mixture of 5 volumes of
obtained with the test solution corresponds to the peak due methanol and 95 volumes of a buffer prepared by
to lamivudine in the chromatogram obtained with the reference dissolving 1.9 g of ammonium acetate in 1000 ml of
solution. water and adjusting the pH to 3.8 ± 0.2 with glacial
C. Melting range (2.4.21). 172° to 178°. acetic acid,
Tests – spectrophotometer set at 270 nm,
Specific optical rotation (2.4.22). –136° to –144°, determined
column efficiency determined from the lamivudine peak is not
Related substances. Determine by liquid chromatography less than 5000 theoretical plates, the tailing factor is not more
(2.4.14), as described in the Assay but using the following than 2.0 and the relative standard deviation for replicate
solutions. injections is not more than 2.0 per cent.
Test solution. Dissolve 0.1 g of the substance under Inject alternatively the test solution and the reference solution.
examination in 100 ml mobile phase.
Calculate the content of C8H11N3O3S.
each of lamivudine RS and salicylic acid in the mobile phase.
Inject the reference solution and record the chromatogram for
at least twice the retention time of lamivudine. The order of
elution is lamivudine and then, salicylic acid. The test is not
Lamivudine Tablets
valid unless the resolution between the peaks due to Lamivudine Tablets contain not less than 90.0 per cent and
lamivudine and salicylic acid is not less than 10, the column not more than 110.0 per cent of the stated amount of
efficiency determined from lamivudine peak is not less than lamivudine, C8H11N3O3S. The tablets may be coated.
5000 theoretical plates and the tailing factor is not more than
1.5. Identification
Separately inject the test solution and the reference solution A. When examined in the range 200 nm to 400 nm (2.4.7), a
and measure the peak responses for each impurity. In the 0.001 per cent w/v solution in a mixture of 50 volumes of water
chromatogram obtained with the test solution, the area of any and 50 volumes of acetonitrile, shows an absorption maximum
peak other than the principal peak is not greater than half of at about 270 nm.
the area of the principal peak in the chromatogram obtained B. In the Assay, the principal peak in the chromatogram
with the reference solution (0.5 per cent) and the sum of the obtained with the test solution corresponds to the peak due
areas of all such peaks is not greater than the area of the to lamivudine in the chromatogram obtained with the reference
principal peak in the chromatogram obtained with the reference solution.
solution (1.0 per cent).
Heavy metals (2.3.13). 1.0 g complies with the limit test for Tests
heavy metals, Method B (20 ppm). Related substances. Determine by liquid chromatography
Sulphated ash (2.3.18). Not more than 0.2 per cent. (2.4.14).
Water (2.3.43). Not more than 0.5 per cent determined on 2.0 g. Test solution. Shake a quantity of the powdered tablets with a
suitable quantity of water, and disperse with the aid of
ultrasound. Add a quantity of acetonitrile containing half of
Test solution. Dissolve 25 mg of the substance under the final volume to get a final concentration of 0.6 per cent
examination in 100 ml of mobile phase. w/v of lamivudine. Mix with the aid of ultrasound for
Reference solution. A 0.025 per cent w/v solution of 10 minutes and make up the volume with water. Filter through
lamivudine RS in mobile phase. a membrane filter disc with an average pore diameter not greater
than 1.0 µm, rejecting the first few ml of the filtrate.
lamivudine RS in a mixture of 80 volumes of water and
20 volumes of acetonitrile.
(5 µm),
– temperature column 35°, Chromatographic system as described under Assay
659
LAMIVUDINE AND TENOFOVIR TABLETS IP 2007
Inject the reference solution. The test is not valid unless the Separately inject the test solution and the reference solution
column efficiency determined from the lamivudine peak is not and measure the peak responses of the major peak.
less than 3000 theoretical plates and the tailing factor is not Calculate the content of C8H11N3O3S in the tablets.
more than 2.0.
Separately inject the test solution. Calculate the amount of
related substances by the area normalisation method. The
content of any individual impurity is not more than 1.0 per
cent and the sum of all impurities is not more than 2.0 per cent. Lamivudine and Tenofovir Tablets
Dissolution (2.5.2). Lamivudine and Tenofovir Disoproxil Fumarate Tablets
Apparatus. No 2
Lamivudine and Tenofovir Tablets contain not less than
Medium. 900 ml of 0.01 M hydrochloric acid
Speed and time. 50 rpm and 30 minutes. amounts of lamivudine, C8H11N3O3S and tenofovir disoproxil
Withdraw a suitable volume of the medium and filter through fumarate, C19H30N5O10P,C4H4O4.
greater than 1.0 µm, rejecting the first 1 ml of the filtrate. Identification
Measure the absorbance of the filtrate, suitably diluted if In the Assay, the principal peak in the chromatogram obtained
necessary, at the maximum at about 270 nm (2.4.7). Calculate with the test solution corresponds to the peak in the
the content of C8H11N3O3S, in the medium from the absorbance chromatogram obtained with the reference solution.
obtained from a solution of known concentration of lamivudine
RS in 0.01 M hydrochloric acid. Tests
D. Not less than 70 per cent of the stated amount of Dissolution (2.5.2).
C8H11N3O3S.
Apparatus. No 1
Other tests. Comply with the tests stated under Tablets. Medium. 900 ml of 0.1 M hydrochloric acid.
Assay. Determine by liquid chromatography (2.4.14). Speed and time. 50 rpm and 30 minutes.
Test solution. Weigh and powder 20 tablets. Shake a quantity Withdraw a suitable volume of the medium and filter.
of the powder containing about 100 mg of Lamivudine with a Determine by liquid chromatography (2.4.14)
mixture of 50 volumes of water and 50 volumes of acetonitrile
to have a final concentration of 0.024 per cent of Lamivudine. Test solution. The filtrate obtained as given above. Dilute the
Disperse with the aid of ultrasound and filter through a filtrate if necessary, with the dissolution medium.
membrane filter disc with an average pore diameter not greater Reference solution. 5.0 ml of each of a 0.16 per cent w/v
than 1.0 µm, rejecting the first few ml of the filtrate. solution of lamivudine RS and of tenofovir disoproxil
Reference solution. A 0.024 per cent w/v solution of fumarate RS in methanol, diluted to 25 ml with the dissolution
lamivudine RS in the same solvent mixture. medium.
Chromatographic system Use the chromatographic system given in the Assay.
– a stainless steel column 15 cm x 4.6 mm, packed with Inject the reference solution. The resolution between the peaks
octadecylsilane chemically bonded to porous silica due to lamivudine and tenofovir disoproxil is not less than 2.0.
(5 µm),
– mobile phase: a degassed mixture of 5 volumes of
methanol and 95 volumes of a buffer prepared by Calculate the contents of C 8 H 11 N 3 O 3 S and
dissolving 1.0 g of ammonium acetate and 1.0 ml of C19H30N5O10P,C4H4O4.
glacial acetic acid in sufficient water to make 1000 ml D. Not less than 75 per cent of the stated amounts of
and adjusting the pH to 3.8 with glacial acetic acid, C8H11N3O3S and C19H30N5O10P,C4H4O4.
– spectrophotometer set at 270 nm, Related substances. Determine by liquid chromatography
– a 10 µl loop injector. (2.4.14).
Inject the reference solution. The test is not valid unless the NOTE — Prepare the solutions immediately before use.
column efficiency determined from the lamivudine peak is not Test solution. Weigh and powder 20 tablets. Weigh accurately
less than 3000 theoretical plates and the tailing factor is not a quantity of the powder containing 100 mg of Lamivudine,
more than 2.0. disperse in 100 ml of mobile phase A and filter.
660
IP 2007 LAMIVUDINE AND ZIDOVUDINE TABLETS
Reference solution (a). A solution containing 0.1 per cent Assay. Determine by liquid chromatography (2.4.14).
w/v of lamivudine RS and tenofovir disoproxil fumarate RS
equivalent to 0.1 per cent w/v of tenofovir disoproxil in mobile
phase A. Test solution. Weigh and powder 20 tablets. Weigh accurately
a quantity of the powder containing 20 mg of Lamivudine,
Reference solution (b). Dilute 1 ml of the solution to 100 ml
dissolve in 100 ml of the mobile phase and filter.
with mobile phase A.
Reference solution (c). A 0.02 per cent w/v solution of fumaric Reference solution. A 0.1 per cent w/v solution each of
acid in mobile phase A. lamivudine RS and tenofovir disoproxil fumarate RS in the
mobile phase. Dilute 20.0 ml of the solution to 100.0 ml with
Chromatographic system the mobile phase.
– a stainless steel column 15 cm x 4.6 mm, packed
octadecylsilane bonded to porous Chromatographic system
silica (5 µm), – a stainless steel column 15 cm x 4.6 mm, packed with
– column temperature 35º, octadecylsilane bonded to porous silica ( 5 µm),
– mobile phase: A. a mixture of 95 volumes of a buffer – column temperature 35º,
solution prepared by dissolving 1.9 g of ammonium – mobile phase: a mixture of 50 volumes of a buffer solution
acetate in 1000 ml of distilled water and adjusting the prepared by dissolving 7.8 g of sodium dihydrogen
pH to 3.8 with glacial acetic acid and 5 volumes of orthophosphate dihydrate in 1000 ml of distilled water,
methanol, adding 1 ml of triethylamine and adjusting the pH to
B. methanol, 2.3 with orthophosphoric acid, and 50 volumes of
– flow rate. 1 ml per minute, acetonitrile,
– a linear gradient programme using the conditions given – flow rate. 1 ml per minute,
below, – spectrophotometer set at 260 nm,
– a 10 µl loop injector. Inject the reference solution. The test is not valid unless the
Time Mobile phase A Mobile phase B column efficiency determined from the peaks due to lamivudine
(in min.) (per cent v/v) (per cent v/v) and tenofovir disoproxil is not less than 750 and 1500
0 100 0 theoretical plates respectively, the tailing factor is not more
than 2.0 and the relative standard deviation for replicate
30 100 0
injections is not more than 2.0 per cent for each component.
31 80 20
35 50 50
60 50 50 Calculate the content of C8H11N3O3S and C19H30N5O10P,C4H4O4
in the tablets.
61 80 20
65 100 0 Storage. Store protected from moisture, at temperature not
exceeding 30º.
67 100 0
column efficiency determined from lamivudine and tenofovir
disoproxil peaks is not less than 750 and 1500 theoretical plates
respectively and the tailing factor is not more than 2.0. Lamivudine and Zidovudine Tablets
Inject the test solution, reference solutions (b) and (c). In the Lamivudine and Zidovudine Tablets contain not less than
chromatogram obtained with the test solution, the area of any 90.0 per cent and not more than 110.0 per cent of the stated
secondary peak is not more than 3.5 times the area of any peak amounts of lamivudine, C 8H 11N 3O 3S, and zidovudine,
in the chromatogram obtained with the reference solution (b) C10H13N5O4. The tablets may be coated.
is not more than 6 times the area of any peak in the Identification
chromatogram obtained with the reference solution (b) (6.0 In the Assay, the two principal peaks in the chromatogram
per cent). Ignore the peak corresponding to the peak in the obtained with the test solution correspond to the peaks due
chromatogram obtained with reference solution (c). to lamivudine and zidovudine in the chromatogram obtained
Other tests. Comply with the tests stated under Tablets. with the reference solution.
661
LAMIVUDINE AND ZIDOVUDINE TABLETS IP 2007
Tests Reference solution (a). A 0.015 per cent w/v solution of

salicylic acid in methanol.
Apparatus. No 2
w/v each of thymine and β-thymidine in methanol.
Medium. 900 ml of 0.1 M hydrochloric acid
Speed and time. 50 rpm and 30 minutes. Reference solution (c). Transfer 5 ml of reference solution (a)
and 15 ml of reference solution (b) to a 100-ml volumetric flask
Withdraw a suitable volume of the medium and filter through and dilute to volume with methanol.
greater than 1.0 µm, rejecting the first 1 ml of the filtrate. Reference solution (d). Weigh accurately about 300 mg of
zidovudine RS and about 150 mg of lamivudine RS, transfer
Determine by liquid chromatography (2.4.14). to a 100-ml volumetric flask, add 30 ml of reference solution (b)
Test solution. Dilute 5.0 ml of the filtrate to 50.0 ml with the and 20 ml of reference solution (a) and disperse with the aid of
mobile phase. ultrasound for about 15 minutes to dissolve. Dilute to volume
with methanol and filter through a membrane filter disc with
Reference solution. Weigh accurately about 30 mg of
an average pore diameter not greater than 1.0 µm, rejecting the
lamivudine RS and 60 mg of zidovudine RS, transfer into a
first few ml of the filtrate.
100-ml volumetric flask, dissolve and dilute to volume with
the mobile phase. Dilute 5.0 ml of this solution to 50.0 ml with Chromatographic system
0.1 M hydrochloric acid. Further dilute 5.0 ml of this solution – a stainless steel column 25 cm x 4.6 mm, packed with
to 20.0 ml with the mobile phase. octadecylsilane chemically bonded to porous silica
(5 µm),
– temperature. column 35°,
– mobile phase: degassed mixtures of methanol and a
buffer prepared by dissolving 1.945 g of ammonium
(5 µm),
acetate in 900 ml of water, adjusting the pH of the
– mobile phase: a degassed mixture of 60 volumes of a
solution to 3.8 with glacial acetic acid and diluting to
buffer solution containing 0.1 M ammonium acetate in
1000.0 ml with water in the proportions and at the
0.1 per cent acetic acid and 40 volumes of methanol,
intervals given below
below,
Inject the reference solution. The test is not valid unless the – spectrophotometer set at 270 nm,
relative standard deviation for replicate injections for each of – a 10 µl loop injector.
the peaks corresponding to lamivudine and zidovudine is not
Time Buffer (pH 3.8) Methanol
(in min.) ( per cent v/v) ( per cent v/v)
Separately inject the test solution and the reference solution 0 90 10
and measure the responses of the major peaks due to
lamivudine and zidovudine. Calculate the contents of 10 90 10
C8H11N3O3S and C10H13N5O4. 25 80 20
D. Not less than 75.0 per cent of the stated amounts of 40 90 10
C8H11N3O3S and C10H13N5O4. 45 90 10
Related substances. Determine by liquid chromatography Inject reference solution (d). The test is not valid unless the
(2.4.14). tailing factor of the lamivudine and zidovudine peaks is not
Test solution. Weigh accurately a quantity of the powdered injections for each of the peaks corresponding to lamivudine
tablets containing the average weight of one tablet, transfer and zidovudine is not more than 2 per cent.
to a 100-ml volumetric flask and add about 50 ml of methanol.
Disperse with the aid of ultrasound for about 10 minutes with Separately inject the test solution and record the
occasional shaking to obtain a uniform dispersion, cool to chromatograms for at least two times the retention time of the
room temperature, dilute to volume with methanol and mix. zidovudine peak. Separately inject reference solutions (a), (b)
Filter through a membrane filter disc with an average pore and (c).
diameter not greater than 1.0 µm, rejecting the first few ml of Calculate the amounts of the related substances by the area
the filtrate. normalisation method. The content of thymine is not greater
662
IP 2007 LAMIVUDINE, NEVIRAPINE AND STAVUDINE DISPERSIBLE TABLETS
than 2.0 per cent, of b-thymidine is not greater than 1.0 per Identification
cent, of salicylic acid is not greater than 0.2 per cent and of
any unknown impurity not greater than 0.5 per cent. The sum In the Assay, the principal peaks in the chromatogram obtained
of all the impurities is not greater than 3.0 per cent. with the test solution correspond to the peaks in the
Assay. Determine by liquid chromatography (2.4.14). Tests
Test solution. Weigh and powder 20 tablets. Transfer an Dissolution (2.5.2).
accurately weighed quantity of the powder equivalent to the Apparatus. No 1
average weight of one tablet into a 200-ml volumetric flask. Medium. 900 ml of 0.1 M hydrochloric acid.
Add 100 ml of the mobile phase and disperse with the aid of Speed and time. 75 rpm and 30 minutes.
ultrasound for about 15 minutes with occasional shaking to
obtain a uniform dispersion. Cool to room temperature and
dilute to volume with the mobile phase. Filter the solution Determine by liquid chromatography (2.4.14).
through a membrane filter disc with an average pore diameter Solvent mixture. 50 volumes of water and 50 volumes of
not greater than 1.0 µm, rejecting the first few ml of the filtrate. methanol.
Dilute 5.0 ml of the filtrate to 25.0 ml with the mobile phase.
Further dilute 5.0 ml to 50.0 ml with the mobile phase and mix. Test solution. The filtrate obtained as given above, diluted
suitably if necessary, with the dissolution medium.
lamivudine RS and 60 mg of zidovudine RS, transfer to a 100- Reference solution. A solution containing 0.02 per cent w/v of
ml volumetric flask, dissolve in the mobile phase and dilute to stavudine RS, 0.09 per cent w/v lamivudine RS and 0.15 per
volume with the mobile phase. Further dilute 5.0 ml of this cent w/v of nevirapine RS dissolved in minimum quantity of
solution to 100.0 ml with the mobile phase. methanol and makeup with solvent mixture. Dilute 5 ml of the
solution to 100 ml with the dissolution medium.
octadecylsilane chemically bonded to porous silica – a stainless steel column 15 cm x 4.6 mm, packed with
(5 µm), octadecylsilane bonded to porous silica (5 µm),
– mobile phase: a degassed mixture of 60 volumes of a – mobile phase: a mixture of 65 volumes of a buffer solution
buffer solution containing 0.1 M ammonium acetate in prepared by dissolving 1 g of octane sulphonic acid
0.1 per cent acetic acid and 40 volumes of methanol, sodium salts and 0.68 g of potassium dihydrogen
– flow rate. 1 ml per minute, phosphate in
– spectrophotometer set at 270 nm, 1000 ml of water, adding 1 ml of triethylamine and
– a 20 µl loop injector. adjusting the pH to 2.5 with orthophosphoric acid, and
relative standard deviation for replicate injections for each of
the peaks corresponding to lamivudine and zidovudine is not
Separately inject the test solution and the reference solution
tailing factor is not more than 1.5 for each component, the
and measure the peak responses of the major peaks.
column efficiency in not less than 2000 theoretical plates for
Calculate the contents of C8H11N3O3S and C10H13N5O4 in the lamivudine and the relative standard deviation for replicate
tablets. injections is not more than 2.0 per cent for each component.
Storage. Store protected from moisture. Inject the test solution and the reference solution.
Calculate the content of C 8H 11N 3 O 3S, C 15H 14 N 4 O and
Lamivudine, Nevirapine and Stavudine C10H12N2O4.
Dispersible Tablets D. Not less than 80 per cent of the stated amounts of
C8H11N3O3S, C15H14N4O and C10H12N2O4.
Lamivudine, Nevirapine And Stavudine Dispersible Tablets
contain not less than 90.0 per cent and not more than 110.0 per
(2.4.14).
cent of the stated amounts of lamivudine, C8H11N3O 3S,
nevirapine, C15H14N4O and stavudine, C10H12N2O4. NOTE — Prepare the solutions immediately before use.
663
LAMIVUDINE, NEVIRAPINE AND STAVUDINE TABLETS IP 2007
Solvent mixture. 70 volumes of 0.2 per cent v/v ortho- Solvent mixture. 50 volumes of water and 50 volumes of
phosphoric acid and 30 volumes of methanol. methanol.
Test solution. Weigh and powder 20 tablets. Weigh accurately Test solution. Weigh accurately a quantity of the powdered
a quantity of the powder containing 75 mg of Lamivudine, tablets containing 50 mg of Lamivudine, dissolve in 100.0 ml
dissolve in 100 ml of the solvent mixture and filter. of solvent mixture. Dilute 10.0 ml of the solution to 50.0 ml with
the solvent mixture and filter.
lamivudine RS, 130 mg of nevirapine RS and 20 mg of stavudine Reference solution. A solution containing 0.10 per cent w/v
RS, dissolve in 20 ml of methanol and dilute to 100 ml with the of lamivudine RS, 0.175 per cent w/v nevirapine RS and 0.025
solvent mixture. Dilute 5 ml of the solution to 50 ml with the per cent w/v of stavudine RS dissolved in minimum quantity
solvent mixture. of methanol and makeup with solvent mixture. Dilute
5.0 ml of the solution to 50.0 ml with the solvent mixture.
Reference solution (b). Dissolve 0.38 mg of Thymine and
0.15 mg of Carboxylic acid in 50 ml of the solvent mixture, add Chromatographic system
10 ml of reference solution (a), dilute to 100 ml with the solvent – a stainless steel column 15 cm x 4.6 mm, packed with
mixture and filter. octadecylsilane bonded to porous silica (5 µm),
Chromatographic system prepared by dissolving 1 g of octane sulphonic acid
– a stainless steel column 25 cm x 4.6 mm, packed with and 0.68 g of potassium dihydrogen phosphate in
octadecylsilane bonded to porous silica (5 µm), 1000 ml of water, adding 1 ml of triethylamine and
– mobile phase: A. methanol, adjusting the pH to 2.5 with orthophosphoric acid, and
B. a buffer solution prepared by 35 volumes of methanol.
dissolving 1.925 g of ammonium acetate in – flow rate. 1 ml per minute.
1000 ml of water and adjusting the pH to 3.0 with – spectrophotometer set at 266 nm,
trifluoroacetic acid, – a 10 µl loop injector.
– a linear gradient programme using the conditions given Inject the reference solution. The test is not valid unless the
below, tailing factor is not more than 1.5 for each component, the
– spectrophotometer set at 266 nm, column efficiency in not less than 2000 theoretical plates for
– a 10 µl loop injector. lamivudine and the relative standard deviation for replicate
injections is not more than 2.0 per cent for each component.
Time Methanol Buffer solution pH 3
(min.) ( per cent) ( per cent) Inject the test solution and the reference solution.
0 5 95 Calculate the content of C 8H 11 N 3 O 3S, C 15H 14 N 4 O and
15 15 85 C10H12N2O4 in the tablets.
30 45 55 Storage. Store protected from moisture, at a temperature not
40 60 40 exceeding 25°.
48 60 40
51 5 95
Lamivudine, Nevirapine and Stavudine
60 5 95
Tablets
Inject reference solution (b). The relative retention times for
carboxylic acid is 0.36, for thymine is 0.42, for lamivudine, 0.74, Lamivudine, Nevirapine and Stavudine Tablets contain not
for stavudine, 1.0 and for nevirapine, 2.19. The test is not valid less than 90.0 per cent and not more than 110.0 per cent of the
unless the tailing factor is not more than 1.5 and the resolution stated amounts of lamivudine, C8H11N 3O3S, nevirapine,
between carboxylic acid and thymine is not less than 2.0. C15H14N4O, and stavudine, C10H12N2O4. The tablets may be
coated.
Inject the test solution. Any individual impurity each for
lamivudine and nevirapine is not more than 1 per cent and for Identification
stavudine, is not more than 3 per cent and the sum of all
impurities is not more than 5.0 per cent. In the Assay, the three principal peaks in the chromatogram
obtained with the test solution have retention times similar to
those of the peaks due to lamivudine, nevirapine and stavudine
Assay. Determine by liquid chromatography (2.4.14). in the chromatogram obtained with the reference solution.
664
IP 2007 LAMIVUDINE, NEVIRAPINE AND STAVUDINE TABLETS
Tests 100 mg of nevirapine, transfer to a 200-ml volumetric flask and

add about 150 ml of water. Disperse with the aid of ultrasound
Dissolution (2.5.2). for 10 minutes with occasional shaking to obtain a uniform
Apparatus. No 2 dispersion, cool to room temperature, dilute to volume with
Medium. 900 ml of 0.01 M hydrochloric acid water and mix. Filter through a membrane filter disc with an
Speed and time. 50 rpm and 45 minutes. average pore diameter not greater than 1.0 µm, rejecting the
a membrane filter disc having an average pore diameter not Reference solution (a). A solution containing 0.15 per cent
greater than 1.0 µm, rejecting the first 1 ml of the filtrate. w/v of stavudine RS and 0.015 per cent w/v of thymine RS in
water.
Reference solution (b). Weigh accurately about 75 mg of
Test solution. Use the filtrate as the test solution. lamivudine RS and about 100 mg of nevirapine RS, transfer
Reference solution. Weigh accurately about 150 mg of to a 200-ml volumetric flask, add 20 ml of methanol and mix
lamivudine RS, 200 mg of nevirapine RS and 30 mg of stavudine with the aid of ultrasound to dissolve. Add 10 ml of the test
RS (if claim of stavudine per tablet is 30 mg) or 40 mg of solution to this solution and make up to volume with water
stavudine RS (if claim of stavudine per tablet is 40 mg) and and filter.
transfer to a 100-ml volumetric flask. Add about 20 ml of Chromatographic system
methanol, disperse with the aid of ultrasound to dissolve and – a stainless steel column 25 cm x 4.6 mm, packed with
dilute to volume with a solvent mixture of equal volumes of octylsilane chemically bonded to porous silica (5 µm),
methanol and water. Dilute 5.0 ml of this solution to 50.0 ml – mobile phase: degassed gradient mixtures of 0.1 M
with 0.01 M hydrochloric acid. ammonium acetate and acetonitrile.
Chromatographic system – flow rate. 1 ml per minute,
– a stainless steel column 15 cm x 4.6 mm, packed with – a linear gradient programme using the conditions given
octadecylsilane chemically bonded to porous silica below,
(5 µm), – spectrophotometer set at 270 nm,
– mobile phase: a degassed mixture of 35 volumes of – a 20 µl loop injector.
methanol and 65 volumes of a buffer prepared by Time 0.1 M Ammonium acetate Acetonitrile
dissolving 0.68 g of potassium dihydrogen phosphate (in min.) (per cent v/v) (per cent v/v)
and 1.0 g of sodium octanesulphonate in 1000.0 ml of 0 95 05
water to which 1 ml of triethylamine is added and the 05 95 05
pH of which is adjusted to 2.5 with phosphoric acid,
25 20 80
30 20 80
– a 10 µl loop injector. 31 95 05
35 95 05
column efficiency determined from the lamivudine peak is not Separately inject reference solutions (a) and (b). The test is
less than 2000 theoretical plates, the tailing factor for the not valid unless the column efficiency determined for the
individual lamivudine, nevirapine and stavudine peaks is not thymine, stavudine, lamivudine and nevirapine peaks is not
more than 1.5 and the relative standard deviation for replicate less than 3000 theoretical plates and the tailing factor for the
injections of all the analyte peaks is not more than 1.0 per cent. same peaks is not more than 2.0.
Separately inject the test solution and measure the peak
Separately inject the test solution and measure the peak
responses of the major peaks due to lamivudine, nevirapine
responses of the major peaks due to lamivudine, nevirapine
and stavudine. Calculate the amounts of related substances
and stavudine. Calculate the contents of C 8H 11N 3O 3 S,
by the area normalisation method. The content of thymine is
C15H14N4O, and C10H12N2O4 respectively.
not greater than 3.0 per cent and that of any other impurity is
D. Not less than 70 per cent of the stated amounts of not greater than 1.0 per cent. The sum of all the impurities is
C8H11N3O3S, C15H14N4O and C10H12N2O4. not greater than 3.5 per cent.
Related substances. Determine by liquid chromatography Other tests. Comply with the tests stated under Tablets.
(2.4.14). Assay. Determine by liquid chromatography (2.4.14).
Test solution. Weigh accurately a quantity of the powdered Test solution. Weigh and powder 20 tablets. Transfer an
tablets (a minimum of 2 tablets should be powdered) containing accurately weighed quantity of the powder containing about
665
LAMIVUDINE ORAL SOLUTION IP 2007
150 mg of lamivudine to a 100-ml volumetric flask, add 20 ml of The principal spot in the chromatogram obtained with the test
methanol and about 50 ml of a mixture of equal volumes of solution corresponds to that in the chromatogram obtained
water and methanol and disperse with the aid of ultrasound with the reference solution.
for 5 minutes. Dilute suitably with the same solvent mixture to B. In the Assay, the principal peak in the chromatogram
obtain a solution containing 0.15 mg of lamivudine per ml. obtained with the test solution corresponds to the peak due
Filter this solution through a membrane filter disc with an to lamivudine in the chromatogram obtained with the reference
average pore diameter not greater than 1.0 µm, rejecting the solution.
Reference solution. A solution containing 0.015 per cent w/v Tests
of lamivudine RS and 0.02 per cent w/v of nevirapine RS and
pH (2.4.24). 5.0 to 7.0.
a concentration of stavudine RS similar to that of the
concentration of stavudine in the test solution. Related substances. Determine by liquid chromatography
(2.4.14).
The chromatographic procedure may be carried out using the
conditions described under Dissolution. Test solution. Transfer an accurately measured volume of the
Inject the reference solution. The test is not valid unless the preparation under examination containing 50 mg of lamivudine
column efficiency determined from the lamivudine peak is not to a 50-ml volumetric flask, add about 30 ml of a solution
less than 2000 theoretical plates, the tailing factor for the prepared by mixing 10 volumes of acetonitrile and 90 volumes
individual peaks due to lamivudine, nevirapine and stavudine of 0.1 M potassium dihydrogen phosphate, the pH of which
is not more than 1.5 and the relative standard deviation for is adjusted to 3.0 with dilute phosphoric acid (solution A),
replicate injections of all the analyte peaks is not more than mix with the aid of ultrasound for 5 minutes, dilute to volume
1.0 per cent. with solution A and filter. Dilute 5 ml of the filtrate to 50 ml with
solution A.
Inject test solution and the reference solution.
Calculate the contents of C 8H 11N3O 3S, C 15H 14N 4O and lamivudine RS and transfer to a 50-ml volumetric flask, dissolve
C10H12N2O4 in the tablets. and dilute to volume with solution A. Further dilute 5 ml of
Storage. Store protected from moisture. this solution to 250 ml with solution A.
Lamivudine Oral Solution octadecylsilane chemically bonded to porous silica
(5 µm),
Lamivudine Oral Solution is a solution of Lamivudine in a
– mobile phase: degassed gradient mixtures of acetonitrile
suitable flavoured vehicle.
and 0.05 M potassium dihydrogen phosphate, adjusting
Lamivudine Oral Solution contains not less than 90.0 per cent the pH of the solution to 3.0 with dilute phosphoric acid,
and not more than 110.0 per cent of the stated amount of – flow rate. 0.8 ml per minute,
lamivudine, C8H11N3O3S. – a linear gradient programme using the conditions given
below,
A. Determine by thin-layer chromatography (2.4.17), coating – a 20 µl loop injector.
the plate with silica gel GF254. Time Phosphate buffer (pH 3.0) Acetonitrile
(in min.) (per cent v/v) ( per cent v/v)
30 volumes of heptane, 30 volumes of acetone and 10 volumes 0 98 02
of strong ammonia solution. 10 85 15
Test solution. Dilute the preparation under examination with 25 30 70
methanol to obtain a solution containing 2 mg of lamivudine 35 30 70
per ml. 40 98 02
Reference solution. A 0.2 per cent w/v solution of lamivudine 50 98 02
RS in a mixture of 75 volumes methanol and 25 volumes of Inject the reference solution. The test is not valid unless the
water. column efficiency determined from the lamivudine peak is not
Apply to the plate 5 µl of each solution. After development, less than 15,000 theoretical plates and the tailing factor is not
dry the plate in air and examine in ultraviolet light at 254 nm. more than 1.5.
666
IP 2007 LAMOTRIGINE
Inject solution A and the test solution. Examine the Lamotrigine is 6-(2,3-dichlorophenyl)-1,2,4-triazine-3,5-
chromatogram obtained with solution A for any extraneous diamine.
peaks and ignore the corresponding peaks observed in the Lamotrigine contains not less than 98.5 per cent and not more
chromatogram obtained with the test solution. Ignore any than 101.5 per cent of C9H7Cl2N5, calculated on the dried basis.
peaks due to preservatives also.
Description. A white to off - white powder.
with the test solution should not be more than 1.0 per cent Identification
and the sum of the areas of all the secondary peaks should
not be more than 2.0 per cent when calculated by percentage Test A may be omitted if tests B and C are carried out. Tests B
area normalisation. and C may be omitted if test A is carried out.
Other tests. Comply with the tests stated under Oral Liquids. A. Determine by infrared absorption spectrophotometry (2.4.6).
Assay. Determine by liquid chromatography (2.4.14). Compare the spectrum with that obtained with lamotrigine
RS or with the reference spectrum of lamotrigine.
Test solution. Weigh accurately a quantity of the preparation
under examination containing about 100 mg of lamivudine in a B. When examined in the range 210 to 360 nm (2.4.7), a 0.0025
100-ml volumetric flask, add about 50 ml of water, mix with the per cent w/v solution in methanol exhibits a maximum at about
aid of ultrasound for 10 minutes, dilute to volume with water, 309 nm.
mix and filter. Dilute 5.0 ml of the filtrate to 50.0 ml with water. C. In the Assay, the chromatogram obtained with test solution
Reference solution. Weigh accurately about 100 mg of corresponds to the chromatogram obtained with reference
lamivudine RS in a 100-ml volumetric flask, dissolve and dilute solution.
to volume with water. Dilute 5.0 ml of this solution to 50.0 ml Tests
with water.
Chromatographic system Related substances. Determine by liquid chromatography
– a stainless steel column 15 cm x 4.6 mm, packed with (2.4.14).
octadecylsilane chemically bonded to porous silica Solvent mixture. 70 volumes of phosphate buffer pH 7.5 and
(5 µm), 30 volumes of acetonitrile.
– mobile phase: 80 volumes of water and 20 volumes of Test solution. Dissolve 100 mg of the substance under
methanol, examination in 100 ml of solvent mixture.
Reference solutionn (a). A 0.05 per cent w/v solution of
lamotrigine RS in solvent mixture.
column efficiency determined from the lamivudine peak is not
less than 3000 theoretical plates, the tailing factor is not more Chromatographic system
than 2.0 and the relative standard deviation for the replicate – a stainless steel column 25 cm x 4.6 mm packed with
injections is not more than 2.0 per cent. octadecylsilane bonded to porous silica (5 µm),
– mobile phase: A. 0.174 per cent w/v solution of
Inject separately the test solution and the reference solution
dipotassium hydrogen phosphate adjusted to pH 7.5
and measure the responses for the major peak.
with orthophosphoric acid and filter,
Determine the weight per ml (2.4.29) of the oral solution and B. acetonitrile,
calculate the content of C8H11N3O3S weight in volume. – a linear gradient programme using the conditions given
below,
– flow rate.1.5 ml per minute,
Lamotrigine – spectrophotometer set at 250 nm,
Cl
Cl
(in mins.) (per cent v/v) (per cent v/v)
N 0 80 20
N 5 80 20
15 25 75
H2N N NH2
30 25 75
C9H7Cl2N5 Mol. Wt. 256.1 35 80 20
667
LAMOTRIGINE DISPERSIBLE TABLETS IP 2007
Inject reference solution (b). Test is not valid unless the column Identification
factor is not more than 2.0. In the Assay, the principal peak in the chromatogram obtained
Inject the test solution and reference solution (b). In the chromatogram obtained with the reference solution.
secondary peak is not more than the area of the peak in the Tests
cent) and the sum of areas of all the secondary peaks is not Dissolution (2.5.2).
more than 2 times the area of the peak in the chromatogram Apparatus No. 1
obtained with the reference solution (b) (1.0 per cent). Medium. 900 ml of 0.1 M hydrochloric acid.
Heavy metals (2.3.13). 2.0 g complies with limit test for heavy Speed and time. 50 rpm for 30 minutes.
metals, Method B (10 ppm). Withdraw a suitable volume of the medium and filter.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Determine by liquid chromatography (2.4.14).
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Test solution. The filtrate obtained as given above.
on 1.0 g by drying in an oven at 105º, under vacuum, for
3 hours. Reference solution. A 0.0055 per cent w/v solution of
lamotrigine RS in the dissolution medium.
Test solution. Dissolve 100 mg of the substance under – a stainless steel column 15 cm x 4.6 mm packed with
examination in 100 ml of mobile phase. Dilute10.0 ml of the octadecylsilane bonded to porous ilica (5 µm),
solution to 50.0 ml with mobile phase. – mobile phase: a mixture of 70 volumes of a 0.408 per cent
Reference solution. A 0.02 per cent w/v solution of lamotrigine w/v solution of potassium dihydrogen phosphate
RS in mobile phase. previously adjusted to pH 7.0 with potassium hydroxide,
30 volumes of acetonitrile
and 70 volumes of 0.408 per cent potassium dihydrogen Inject the reference solution. The test is not valid unless the
orthophosphate adjusted the pH to 7.0 with dilute column efficiency is not less than 3000 theoretical plates, the
potassium hydroxide solution. tailing factor is not more than 2.0 and the relative standard
– flow rate.1.2 ml per minute, deviation for replicate injections is not more than 2.0 per cent.
– spectrophotometer set at 308 nm, Inject the test solution and the reference solution.
D. Not less than 75 per cent of the stated amount of C9H7Cl2N5.
column efficiency is not less than 2500 theoretical plates, the Other tests. Comply with the tests stated under Tablets.
tailing factor is not more than 2.0 and the relative standard Assay. Determine by liquid chromatography (2.4.14).
Test solution. Accurately transfer 10 intact tablets in a suitable
Inject the test solution and the reference solution. volumetric flask, add mobile phase, sonicate to dissolve and
Calculate the content of C9H7Cl2N5. dilute with mobile phase to obtain a concentration of 0.02 per
cent w/v of Lamotrigine.
exceeding 25°. Reference solution. A 0.02 per cent w/v solution of lamotrigine
RS in mobile phase.
Lamotrigine Dispersible Tablets octadecylsilane bonded to porous silica (5 µm),
Lamotrigine Dispersible Tablets contain not less than 90.0 per w/v solution of potassium dihydrogen phosphate
cent and not more than 110.0 per cent of the stated amount of previously adjusted to pH 7.0 with potassium hydroxide,
lamotrigine, C9H7Cl2N5. 30 volumes of acetonitrile,
668
IP 2007 LANATOSIDE C
– flow rate. 1.2 ml per minute, A. Determine by infrared absorption spectrophotometry (2.4.6).
– spectrophotometer set at 308 nm, Compare the spectrum with that obtained with lanatoside C
– a 10 µl loop injector. RS.
Inject the reference solution. The test is not valid unless the B. In the test for Related substances, the principal band in the
column efficiency is not less than 2500 theoretical plates, the chromatogram obtained with test solution (b) corresponds to
tailing factor is not more than 2.0 and the relative standard that in the chromatogram obtained with reference solution (a).
C. Suspend about 0.5 mg in 0.5 ml of ethanol (60 per cent)
Inject the test solution and the reference solution. and add 0.1 ml of 3,5-dinitrobenzoic acid solution and 0.1 ml
of 2 M sodium hydroxide; a violet colour develops.
Calculate the content of C9H7Cl2N5.
D. Dissolve 5 mg in 5 ml of glacial acetic acid, add 0.1 ml of
ferric chloride test solution, mix and add 2 ml of sulphuric
Labelling. The label states that the tablets should be dispersed acid slowly so as to form a lower layer; a brown ring is formed
in water immediately before use. at the junction of the two liquids and the upper layer develops
a green colour which becomes blue on standing.
Tests
Lanatoside C Appearance of solution. A 2.0 per cent w/v solution in
than reference solutions YS7 or BYS7 (2.4.1).
CH3
R
O O in a 2.0 per cent w/v solution in methanol.
CH3
O O Related substances. Determine by thin-layer chromatography
OH (2.4.17), coating the plate with silica gel G.
CH3
O O
OH Mobile phase. A mixture of 60 volumes of toluene, 30 volumes
HO
O
of ethanol (95 per cent), 20 volumes of dichloromethane and
O O O
O CH3
1 volume of water.
OH HO
H3C
OH O Test solution (a). Dissolve 0.2 g of the substance under
OH H3C H
R= H OH
H
C49H76O20 Mol. Wt. 985.1 lanatoside C RS in methanol.
Lanatoside C is 3-[(O-β-D-glucopyranosyl-(1→ 4)-O-3- Reference solution (b). A 0.03 per cent w/v solution of
acetyl-2,6-dideoxy-β- D -ribo-hexopyranosyl-(1→ 4)-O-2,6- lanatoside C RS in methanol.
dideoxy-β-D-ribo-hexopyranosyl-(1→4)-)-O-2,6-dideoxy-β-
D-ribo-hexopyranosyl)oxy]-12,14-dihydroxy-3β,5β,2β-card-
20(22)-enolide. lanatoside C RS in methanol.
Lanatoside C contains not less than 97.0 per cent and not Reference solution (d). A 0.01 per cent w/v solution of
more than 103.0 per cent of C49H76O20, calculated on the dried lanatoside C RS in methanol.
basis. Apply to the plate 5 µl of each solution as bands. After
Description. A white or slightly yellowish, crystalline powder development, dry the plate in a current of cool air for 5 minutes
or fine crystals; hygroscopic. and carry out a second development in the same direction.
After development, dry the plate in a current of cool air for
Identification 5 minutes, spray with ethanolic sulphuric acid (5 per cent),
heat at 140° for 15 minutes and examine in daylight. In the
Test A may be omitted if tests B, C and D are carried out. Tests chromatogram obtained with test solution (a) any secondary
B, C and D may be omitted if test A is carried out. band is not more intense than the band in the chromatogram
669
LANATOSIDE C TABLETS IP 2007
obtained with reference solution (b), not more than three such 10 volumes of chloroform, 5 volumes of methanol and
bands are more intense than the band in the chromatogram 1 volume of water, filter, evaporate the filtrate to dryness and
obtained with reference solution (d) and not more than one of dissolve the residue in 5 ml of methanol.
these bands is more intense than the band in the chromatogram References solution (a). A 0.05 per cent w/v solution of
obtained with reference solution (c). lanatoside C RS in methanol.
on the residue from the test for Loss on drying.
with methanol.
Loss on drying (2.4.19). Not more than 7.5 per cent, determined Apply to the plate 2 µl of each solution. Allow the mobile
on 0.5 g by drying over phosphorus pentoxide at 105° at a phase to rise 13 cm. Dry the plate at 100° for 15 minutes, cool
pressure of 1.5 to 2.5 kPa. and develop once again with a mixture of 40 volumes of
Assay. Before carrying out the Assay, allow both the substance 1-propanol, 40 volumes of water, 10 volumes of ethyl acetate
under examination and the reference substance to stand in a and 10 volumes of strong ammonia solution as the mobile
desiccator containing a saturated solution of potassium phase. Allow the mobile phase to rise 13 cm. Dry the plate at
thiocyanate for 24 hours. Weigh accurately about 50 mg, 120° for 1 hour, cool and spray with a 5 per cent w/v solution
dissolve in sufficient ethanol (95 per cent) to produce 50.0 ml of potassium dichromate in a 40 per cent w/w solution of
and dilute 5.0 ml to 100.0 ml with the same solvent. To 5.0 ml of sulphuric acid. Any secondary spot in the chromatogram
this solution add 3 ml of alkaline sodium picrate solution obtained with the test solution is not more intense than the
and allow to stand protected from light in a water-bath at 19° spot in the chromatogram obtained with reference solution (b).
to 21° for 40 minutes. Measure the absorbance of the resulting Uniformity of content. Comply with the test stated under
solution at the maximum at about 484 nm (2.4.7), using as the Tablets.
blank a mixture of 5.0 ml of ethanol (95 per cent) and 3 ml of
alkaline sodium picrate solution prepared at the same time. Powder one tablet, shake with 10.0 ml of ethanol (95 per cent)
for 15 minutes and centrifuge, protecting the mixture from
Calculate the content of C49H76O20 from the absorbance direct sunlight.To 5.0 ml of the clear supernatant liquid, add
obtained by repeating the operation using lanatoside C RS in 3 ml of alkaline sodium picrate solution and allow to stand
place of the substance under examination. protected from light in a water-bath at 19° to 21° for 40 minutes.
Storage. Store protected from light and moisture, in well-filled Measure the absorbance of the resulting solution at the
glass containers in a cold place. maximum at about 484 nm (2.4.7), using as the blank a mixture
of 5.0 ml of ethanol (95 per cent) and 3 ml of alkaline sodium
picrate solution prepared at the same time. Calculate the
content of C 49H 76O 20 from the absorbance obtained by
repeating the operation using lanatoside C RS in place of the
Lanatoside C Tablets substance under examination.
Lanatoside Tablets contain not less than 90.0 per cent and not Other tests. Comply with the tests stated under Tablets.
more than 110.0 per cent of the stated amount of lanatoside C, Assay. Weigh and powder 20 tablets. Weigh accurately a
C49H76O20. quantity of the powder containing about 5 mg of Lanatoside
C and shake with 50 ml of ethanol (95 per cent) in a mechanical
Identification shaker for 15 minutes, protecting the mixture from direct
In the test for Related substances, the principal spot in the sunlight. Dilute to 100.0 ml with ethanol (95 per cent), mix
chromatogram obtained with the test solution corresponds to and filter, discarding the first 20 ml of the filtrate. To 5.0 ml of
that in the chromatogram obtained with reference solution (a). the filtrate add 3 ml of alkaline sodium picrate solution and
allow to stand protected from light in a water-bath at 19° to 21°
Tests for 40 minutes. Measure the absorbance of the resulting
Related substances. Determine by thin-layer chromatography blank a mixture of 5.0 ml of ethanol (95 per cent) and 3 ml of
(2.4.17), coating the plate with silica gel G. alkaline sodium picrate solution prepared at the same time.
Mobile phase. A mixture of 80 volumes of hexane, 10 volumes Calculate the content of C49H76O20 from the absorbance
of ether and 10 volumes of glacial acetic acid. obtained by repeating the operation using lanatoside C RS in
place of the substance under examination.
containing 2.5 mg of Lanatoside C with 10 ml of a mixture of Storage. Store protected from light and moisture.
670
IP 2007 LEVAMISOLE TABLETS
Levamisole Hydrochloride Test solution (a). Dissolve 0.5 g of the substance under
N S Test solution (b). Dissolve 0.5 g of the substance under

H examination in 100 ml of methanol.
N , HCl
C11H12N2S,HCl Mol. Wt. 240.8 levamisole hydrochloride RS in methanol.
Levamisole Hydrochloride is (S)-2,3,5,6-tetrahydro-6- Apply to the plate 10 µl of each solution. After development,
phenylimidazo[2,1-b]thiazole hydrochloride. dry the plate at 105° for 15 minutes and examine in ultraviolet
Levamisole Hydrochloride contains not less than 98.5 per cent light at 254 nm. Any secondary spot in the chromatogram
and not more than 101.0 per cent of C11H12N2S,HCl, calculated obtained with test solution (a) is not more intense than the
on the dried basis. spot in the chromatogram obtained with reference solution
(a). Expose the plate to iodine vapour in an airtight tank for
15 minutes. Any secondary spot in the chromatogram obtained
Identification with test solution (a), other than any spot with a very low
Rf value, is not more intense than the spot in the chromatogram
Test A may be omitted if tests B, C, D and E are carried out. obtained with reference solution (a).
Tests B and D may be omitted if tests A, C and E are carried
out.
Compare the spectrum with that obtained with levamisole RS
or with the reference spectrum of levamisole. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
chromatogram obtained with test solution (b) examined in Assay. Weigh accurately about 0.2 g, dissolve in 30 ml of
ultraviolet light at 254 nm corresponds to that in the ethanol (95 per cent), add 5 ml of 0.01 M hydrochloric acid.
chromatogram obtained with reference solution (b). Titrate with 0.1 M sodium hydroxide determining the end-
point potentiometrically (2.4.25). Record the volume added
C. Complies with the test for Specific optical rotation.
between the two inflections.
D. Dissolve 0.5 g in 20 ml of water and add 6 ml of 1 M sodium
hydroxide. Extract with 20 ml of dichloromethane, wash the
C11H12N2S,HCl.
lower layer with two quantities, each of 10 ml, of water, dry
over anhydrous sodium sulphate, filter and evaporate the Storage. Store protected from light and moisture.
solvent at a temperature not exceeding 40° under reduced
pressure. The residue melts at 58° to 61° (2.4.21).
E. Gives reaction A of chlorides (2.3.1). Levamisole Tablets
Levamisole Hydrochloride Tablets
Tests
Levamisole Tablets contain not less than 90.0 per cent and
Appearance of solution. A 5.0 per cent w/v solution in carbon not more than 110.0 per cent of the stated amount of
dioxide-free water is clear (2.4.1), and not more intensely levamisole, C11H12N2S.
Identification
Specific optical rotation (2.4.22). –121° to –128°, determined In the test for Related substances, the principal spot in the
in a 5.0 per cent w/v solution. chromatogram obtained with test solution (b) corresponds to
(2.4.17), coating the plate with silica gel HF254. Tests
Mobile phase. A mixture of 60 volumes of toluene, 40 volumes Related substances. Determine by thin-layer chromatography
of acetone and 1 volume of strong ammonia solution. (2.4.17), coating the plate with silica gel HF254.
671
LEVOCETIRIZINE HYDROCHLORIDE IP 2007
Mobile phase. A mixture of 60 volumes of toluene, 40 volumes Levocetirizine dihydrochloride is (R)-2-[2-[4-[(4-

of acetone and 1 volume of strong ammonia solution. chlorophenyl)phenylmethyl]piperazin-1-yl]ethoxy]acetic
Test solution (a). Shake a quantity of the powdered tablets acid dihydrochloride.
containing 100 mg of levamisole with 5 ml of methanol for Levocetirizine Hydrochloride contains not less than 98.0 per
2 minutes and filter. cent and not more than 102.0 per cent of C21H25N2O3Cl,2HCl
methanol. Description. A white or almost white powder.
levamisole hydrochloride RS in methanol.
Identification
Referece solution (b). Dilute 1 ml of reference solution (a) to A. Determine by infrared absorption spectrophotometry (2.4.6).
20 ml with methanol. Compare the spectrum with that obtained with levocetirizine
dihydrochloride RS or with the reference spectrum of
Apply to the plate 10 µl of each solution. After development, levocetirizine dihydrochloride.
dry the plate at 105° for 15 minutes and examine in ultraviolet
light at 254 nm as well as after exposure to iodine vapour. Any B. When examined in the range 200 nm and 350 nm (2.4.7), a
secondary spot the chromatogram obtained with test solution 0.001 per cent w/v solution in methanol shows an absorption
(a) is not more intense than the spot in the chromatogram maximum at about 230 nm.
Tests
Specific optical rotation (2.4.22). +10o to +14o, determined in a
1 per cent w/v solution in carbon dioxide-free water at
quantity of the powder containing about 300 mg of levamisole,
365 nm.
add 30 ml of water and shake for 10 minutes. Filter, wash the
filter with 20 ml of water and add the washings to the filtrate. Heavy metals (2.3.13). The residue obtained from Sulphated
To the combined filtrate add dilute ammonia solution to make ash complies with limit test for heavy metals, Method D (20
it alkaline and extract with three quantities each of 25 ml, 15 ml ppm).
and 15 ml, of chloroform, filter through cotton wool covered
Enantiomeric purity. Determine by liquid chromatography
with a layer of anhydrous sodium sulphate. Combine the
(2.4.14).
chloroform extracts and evaporate to dryness. Dissolve the
residue in 50 ml of anhydrous glacial acetic acid. Titrate with Test solution. Dissolve 25 mg of the substance under
0.1 M perchloric acid, using crystal violet solution as examination in 2 ml of methanol and dilute to 50 ml with ethanol
indicator. Carry out a blank titration. ( 95 per cent) and filter.
1 ml of 0.1 M perchloric acid is equivalent to 0.02043 g of Reference solution. Dissolve 25 mg of the racemic cetirizine
C11H12N2S. dihydrochloride RS in 2 ml of methanol and dilute to 50 ml
with ethanol ( 95 per cent).
equivalent amount of levamisole.
chiral Pak AD-H (5 µm),
– mobile phase: a mixture of 70 volumes of n-hexane, 15
volumes of isopropyl alcohol, 15 volumes of ethanol
Levocetirizine Hydrochloride ( 95 per cent) and 0.02 volume of tri-fluoro acetic acid,
Cl O COOH
N – a 20 µl loop injector.
N Inject the reference solution and the test solution.
H , 2HCl
The relative retention time of levocetirizine isomer is about 2
with respect to levocetirizine peak.
Calculate the chiral purity of levocetirizine dihydrochloride
by area normalization method, the enantiomeric purity is not
C21H25N2O3Cl,2HCl Mol. Wt. 461.8 less than 98 per cent.
672
IP 2007 LEVOCETIRIZINE TABLETS
Related substances. Determine by liquid chromatography Identification

(2.4.14).
Test solution. Dissolve 20 mg of the substance under with the test solution corresponds to the peak in the
examination in 100 ml of mobile phase and filter. chromatogram obtained with the reference solution.
levocetrizine dihydrochloride RS in mobile phase. Tests
Reference solution (b). Dilute 1 ml of reference solution (a) to Dissolution (2.5.2).
100 ml with mobile phase. Apparatus No. 1
Chromatographic system Medium. 900 ml of phosphate buffer pH 6.8
– a stainless steel column 25 cm x 4.6 mm packed with Speed and time. 50 rpm for 30 minutes.
octadecysilane bonded to porous silica (5 µm),
– mobile phase: a mixture of 0.4 volume of 5.5 per cent v/
v solution of sulphuric acid, 6.6 volumes of water and Determine by liquid chromatography (2.4.14).
93 volumes of acetonitrile, Test solution. Use the filterate.
– spectrophotometer set at 230 nm, Reference solution. A 0.025 per cent w/v solution of
– a 20 µl loop injector. levocetirizine dihydrochloride RS in mobile phase. Dilute
1 ml of the solution to 50 ml with dissolution medium.
Inject reference solution (b). Test is not valid unless the column
factor is not more than 2.0. Calculate the content of C21H27Cl3N2O3,2HCl.
Inject the test solution and reference solution (b). In the D. Not less than 75 per cent of the stated amount of
chromatogram obtained with the test solution, the area of any C21H27Cl3N2O3,2HCl.
secondary peak is not more than 0.5 times the area of the peak Related substances. Determine by liquid chromatography
in the chromatogram obtained with reference solution (b) (0.5 (2.4.14).
not more than the area of the peak in the chromatogram Test solution. Weigh and powder 20 tablets. Weigh accurately
obtained with the reference solution (b) (1.0 per cent). a quantity of the powdered tablet containing 25 mg of
levocetirizine dihydrochloride, dissolve in 100 ml of mobile
Sulphated ash (2.3.18). Not more than 0.2 per cent. phase.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Reference solutionn (a). A 0.1 per cent w/v solution of
on 1 g by drying in an oven at 100o at a pressure not exceeding lamotrigine RS in mobile phase.
0.7 kPa.
Assay. Weigh accurately about 0.1 g, dissolve in 70 ml of a 100 ml with mobile phase.
mixture of 30 ml of water and 70 ml of acetone. Titrate with 0.1
M sodium hydroxide upto the second point of inflection. Chromatographic system as described under Assay.
Determine the end-point potentiometrically (2.4.25). Carry out Inject reference solution (a). Test is not valid unless the column
a blank titration. effciency is not less than 2000 theoretical plates and the tailing
1 ml of 0.1 M sodium hydroxide is equivalent to 0.01539 g of factor is not more than 2.0.
C21H25N2O3Cl,2HCl. Inject the test solution and reference solution (b). In the
Storage. Store protected from moisture. chromatogram obtained with the test solution, the area of any
more than twice the area of the peak in the chromatogram
Levocetirizine Tablets obtained with the reference solution (b) (2.0 per cent).
Levocetirizine Dihydrochloride Tablets Uniformity of content. Comply with the test stated under
Levocetirizine Tablets contain not less than 90.0 per cent and Tablets.
not more than 110.0 per cent of the stated amount of Determine by liquid chromatography (2.4.14), as described
levocetirizine hydrochloride, C21H27Cl3N2O3,2HCl. under Assay.
673
LEVODOPA IP 2007
Test solution. Transfer 1 tablet in a suitable volumetric flask. Identification

Add about 20 per cent v/v water to disperse with shaking.
Further dilute with methanol to obtain a final concentration of Test A may be omitted if tests B, C and D are carried out. Tests
0.0025 per cent w/v. B, C and D may be omitted if test A is carried out.
Other tests. Comply with the tests stated under Tablets. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with levodopa RS
or with the reference spectrum of levodopa.
a quantity of the powdered tablet containing 25 mg of B. Dissolve about 2 mg in 2 ml of water and add 0.2 ml of ferric
Levocetirizine Dihydrochloride, disperse in 100.0 ml of mobile chloride solution; a green colour develops which changes to
phase and filter. Dilute 5.0 ml of the solution to 25.0 ml with bluish violet on the addition of 0.1 g of hexamine.
mobile phase. C. Dissolve about 5 mg in a mixture of 5 ml of 1 M hydrochloric
Reference solution. A 0.005 per cent w/v solution of acid and 5 ml of water. Add 0.1 ml of sodium nitrite solution
levocetrizine dihydrochloride RS in mobile phase. containing 10 per cent w/v of ammonium molybdate; a yellow
colour develops which changes to red on the addition of
Chromatographic system 10 M sodium hydroxide.
octadecylsilane chemically bonded to porous silica D. Mix about 5 mg with 1 ml of water, 1 ml of pyridine and
(5 µm), about 5 mg of 4-nitrobenzoyl chloride and allow to stand for
– mobile phase: a mixture of 60 volumes of 0.05 M 3 minutes; a violet colour develops which changes to pale
potassium dihydrogen phosphate and 40 volumes of yellow on boiling. Add, while shaking, 0.2 ml of sodium
acetonitrile, adjust the pH to 6.0 with 10 per cent w/v of carbonate solution; the violet colour reappears.
sodium hydroxide,
– flow rate. 1 ml per minute, Tests
hydrochloric acid is not more intensely coloured than
Inject the reference solution. The test is not valid unless the reference solution BYS6 (2.4.1).
tailing factor is not more than 2.0. The column efficiency in
pH (2.4.24). 4.5 to 7.0, determined in a suspension prepared by
not less than 1500 theoretical plates. The relative standard
shaking 0.1 g with 10 ml of carbon dioxide-free water for
15 minutes.
Optical rotation (2.4.22). –1.27° to –1.34°, determined at 20° in
Calculate the content of C21H27Cl3N2O3,2HCl. a solution prepared in the following manner. Dissolve a quantity
Storage. Store protected from light and moisture. containing 0.2 g of the substance on the dried basis and 5 g of
hexamine in 10 ml of 1 M hydrochloric acid, add sufficient
Labelling. The label states the strength of Levocetirizine
1 M hydrochloric acid to produce 25 ml and allow to stand for
Dihydrochloride.
3 hours, protected from light.
Light absorption. Dissolve 30 mg in sufficient 0.1 M
hydrochloric acid to produce 100.0 ml and dilute 10.0 ml to
Levodopa 100.0 ml with 0.1 M hydrochloric acid.
L-Dopa When examined in the range 230 nm to 360 nm (2.4.7), the
HO resulting solution shows an absorption maximum only at about
H NH2 280 nm; absorbance at 280 nm, 0.41 to 0.44.
HO COOH Related substances. Determine by thin-layer chromatography

C9H11NO4 Mol. Wt. 197.2
Levodopa is 3-(3,4-dihydroxyphenyl)-L-alanine.
25 volumes of glacial acetic acid and 25 volumes of water.
Levodopa contains not less than 98.5 per cent and not more
examination in 5 ml of anhydrous formic acid and add
Description. A white or slightly cream, crystalline powder; sufficient methanol to produce 10 ml; prepare immediately
odourless. before use.
674
IP 2007 LEVODOPA AND CARBIDOPA TABLETS
Reference solution (a). Dilute 1 volume of the test solution to due to carbidopa and levodopa in the chromatogram obtained
200 volumes with methanol. with the reference solution.
Reference solution (b). A mixture of equal volumes of the test B. Shake a quantity of the powdered tablets containing 50 mg
solution and a solution prepared by dissolving 30 mg of of Levodopa with 4 ml of ethanol (95 per cent) and 1 ml of
L-tyrosine in 1 ml of anhydrous formic acid and diluting to 1 M sulphuric acid. Add 2 ml of cinnamaldehyde, allow to
100 ml with methanol. stand for 20 minutes, add 50 ml of 0.1 M hydrochloric acid,
Apply to the plate 10 µl each of the test solution and reference shake for 2 minutes and allow to stand. Filter the aqueous
solution (a) and 20 µl of reference solution (b) as bands. Allow layer and to 5 ml of the filtrate add 0.1 ml of ferric chloride test
the mobile phase to rise 20 cm. Dry the plate in a current of air. solution. To half of the solution add an excess of dilute
After development, dry the plate in a current of warm air, spray ammonia solution; a purple colour is produced. To the
with a freshly prepared mixture of equal volumes of a 10 per remainder add an excess of sodium hydroxide solution; a
cent w/v solution of ferric chloride and a 5 per cent w/v deep red colour is produced.
solution of potassium ferricyanide and examine the plate C. Shake a quantity of the powdered tablets containing 1 mg
immediately. Any secondary band in the chromatogram of anhydrous carbidopa with 5 ml of 0.05 M sulphuric acid
obtained with the test solution is not more intense than the and filter. Add 5 ml of dimethylaminobenzaldehyde reagent
band in the chromatogram obtained with reference solution to the filtrate; a yellow colour is produced.
with reference solution (b) shows a distinct band, at a higher Tests
Rf value than the principal band, which is more intense than
Uniformity of Content. For tablets containing 10 mg or less
the band in the chromatogram obtained with reference solution
of Carbidopa.
(a).
Comply with the test stated under Tablets.
heavy metals, Method B (20 ppm). Determine by liquid chromatography (2.4.14).
Sulphated ash (2.3.18). Not more than 0.1 per cent. Test solution. Shake one tablet with 20 ml of 0.1 M phosphoric
Loss on drying (2.4.19). Not more than 1.0 per cent, determined acid for 30 minutes, add sufficient water to produce 200.0 ml,
on 1.0 g by drying in an oven at 105°. mix and filter.
Assay. Weigh accurately about 0.3 g, dissolve, heating if Reference solution. Weigh accurately about 10 mg of
necessary, in 5 ml of anhydrous formic acid and add 25 ml of carbidopa RS, dissolve in 20 ml of 0.1 M phosphoric acid by
anhydrous glacial acetic acid and 25 ml of dioxan. Titrate gentle warming and add sufficient water to produce 200.0 ml.
with 0.1 M perchloric acid, using 0.1 ml of crystal violet Carry out the chromatographic procedure described under
solution as indicator and titrating to a green end-point. Carry Assay.
C9H11NO4.
Storage. Store protected from light and moisture. Assay. Determine by liquid chromatography (2.4.14).
Test solution. Weigh and powder 20 tablets. Shake a quantity
of the powder containing about 250 mg of Levodopa and
25 mg of Carbidopa, with 50 ml of 0.1 M phosphoric acid for
Levodopa and Carbidopa Tablets 30 minutes, add sufficient water to produce 500.0 ml, mix and
Co-careldopa Tablets filter.
Levodopa and Carbidopa Tablets contain not less than Reference solution. Weigh accurately about 250 mg of
90.0 per cent and not more than 110.0 per cent of the stated levodopa RS and about 25 mg of carbidopa RS, dissolve in
amount of anhydrous carbidopa, C10H14N2O4, and not less 2 ml of 0.1 M phosphoric acid by gentle warming and add
than 95.0 per cent and not more than 105.0 per cent of the sufficient water to produce 500.0 ml.
stated amount of levodopa, C9H11NO4. Chromatographic system
Identification octylsilane chemically bonded to porous silica (5 µm),
A. In the Assay, the chromatogram obtained with the test – mobile phase: mix 950 ml of sodium dihydrogen
solution shows two principal peaks that correspond to those phosphate solution (1.162 per cent w/v) with 1.3 ml of
675
LEVODOPA CAPSULES IP 2007
sodium 1-decanesulfonate solution (0.024 per cent of ferric chloride and a 5 per cent w/v solution of potassium
w/v), adjust to a pH of about 2.8 with phosphoric acid ferricyanide. The principal spot in the chromatogram obtained
and dilute with water to produce 1000 ml, with the test solution corresponds to that in the chromatogram
– flow rate. 2 ml per minute, obtained with the reference solution.
– a 20 µl loop injector. Tests
Inject alternately the test solution and the reference solution. Specific optical rotation (2.4.22). –38.5° to –41.5°, determined
The retention times are about 4 minutes and 11 minutes for in the following manner. Weigh accurately a quantity of the
levodopa and carbidopa respectively. contents of the capsules containing 1.25 g of Levodopa, shake
Calculate the content of C10H14N2O4 and C9H11NO4 in the with 25.0 ml of 0.5 M hydrochloric acid for 30 minutes,
tablets. centrifuge and filter the supernatant liquid. To 10.0 ml of the
filtrate add 10 ml of a 21.5 per cent w/v solution of aluminium
Storage. Store protected from light and moisture. sulphate, 20 ml of a 21.8 per cent w/v solution of sodium
Labelling. The label states the quantity of Carbidopa in terms acetate and sufficient water to produce 50.0 ml and measure
of the equivalent amount of anhydrous carbidopa, and the the optical rotation of the resulting solution at 20°. Separately
quantity of Levodopa in each tablet. dilute 5.0 ml of the filtrate to 200.0 ml with 0.1 M hydrochloric
acid, mix well and dilute 10.0 ml to 200.0 ml with 0.1 M
content of C9H11NO4, in the filtrate taking 142 as the specific
Levodopa Capsules absorbance at 280 nm and from the result so obtained calculate
L-Dopa Capsules the specific optical rotation.
Levodopa Capsules contain not less than 95.0 per cent and Related substances. Determine by thin-layer chromatography
not more than 105.0 per cent of the stated amount of levodopa, (2.4.17), coating the plate with microcrystalline cellulose.
C9H11NO4. Mobile phase. A mixture of 50 volumes of 1-butanol, 25
volumes of glacial acetic acid and 25 volumes of water.
Identification
Test solution. A solution prepared immediately before use by
A. Dissolve as completely as possible a quantity of the shaking a quantity of the contents of the capsules containing
contents of the capsules containing 0.5 g of Levodopa in 0.1 g of Levodopa with 10 ml of a mixture of equal volumes of
25 ml of 1 M hydrochloric acid and filter. Adjust the pH to 3 anhydrous formic acid and methanol.
with 5 M ammonia, added dropwise with stirring, and allow to
stand for several hours, protected from light. Filter, wash the Reference solution (a). Dilute 1 volume of the test solution to
precipitate and dry it at 105°. 200 volumes with methanol.
On the residue, determine by infrared absorption Reference solution (b). A mixture of equal volumes of the test
spectrophotometry (2.4.6). Compare the spectrum with that solution and a solution prepared by dissolving 30 mg of L-
obtained with levodopa RS or with the reference spectrum of tyrosine in 1 ml of anhydrous formic acid and diluting to
levodopa. 100 ml with methanol.
B. Determine by thin-layer chromatography (2.4.17), coating Apply to the plate 10 µl of each of the test solution and reference
the plate with microcrystalline cellulose. solution (a) and 20 µl of reference solution (b) as bands. Allow
the mobile phase to rise 20 cm. After development, dry the
Mobile phase. A mixture of 50 volumes of 1-butanol, 25 plate in a current of warm air, spray with a freshly prepared
volumes of glacial acetic acid and 25 volumes of water. mixture of equal volumes of a 10 per cent w/v solution of ferric
Test solution. Shake a quantity of the contents of the capsules chloride and a 5 per cent w/v solution of potassium
containing 0.1 g of Levodopa with 10 ml of 1 M hydrochloric ferricyanide and examine the plate immediately. Any
acid and filter. secondary band in the chromatogram obtained with the test
solution is not more intense than the band in the chromatogram
Reference solution. A 1 per cent w/v solution of levodopa RS
obtained with reference solution (a). The test is not valid unless
in 1 M hydrochloric acid.
the chromatogram obtained with reference solution (b) shows
Apply to the plate 5 µl of each solution. After development, a distinct band, at a higher Rf value than the principal band,
dry the plate in a current of warm air and spray with a freshly which is more intense than the band in the chromatogram
prepared mixture of equal volumes of a 10 per cent w/v solution obtained with reference solution (a).
676
IP 2007 LEVODOPA TABLETS
Dissolution (2.5.2). Reference solution. A 1 per cent w/v solution of levodopa RS

Apparatus. No 2 in 1 M hydrochloric acid.
Medium. 900 ml of 0.1 M hydrochloric acid Apply to the plate 5 µl of each solution. After development,
Speed and time. 75 rpm and 30 minutes. dry the plate in a current of warm air and spray with a freshly
prepared mixture of equal volumes of a 10 per cent w/v solution
Withdraw a suitable volume of the medium and filter. Measure of ferric chloride and a 5 per cent w/v solution of potassium
the absorbance of the resulting solution at the maximum at ferricyanide. The principal spot in the chromatogram obtained
about 280 nm (2.4.7). Calculate the content of C9H11NO4 taking with the test solution corresponds to that in the chromatogram
141 as the specific absorbance at 280 nm. obtained with the reference solution.
D. Not less than 75 per cent of the stated amount of C9H11NO4.
Tests
Assay. Weigh accurately a quantity of the mixed contents of Specific optical rotation (2.4.22). –38.5° to –41.5°, determined
20 capsules containing about 0.4 g of Levodopa, dissolve as in the following manner. Weigh accurately a quantity of the
completely as possible in 10 ml of anhydrous formic acid, add powdered tablets containing 1.25 g of Levodopa, shake with
80 ml of anhydrous glacial acetic acid. Titrate with 0.1 M 25.0 ml of 0.5 M hydrochloric acid for 30 minutes, centrifuge
perchloric acid, using oracet blue B solution as indicator. and filter the supernatant liquid. To 10.0 ml of the filtrate add
Carry out a blank titration. 10 ml of a 21.5 per cent w/v solution of aluminium sulphate,
20 ml of a 21.8 per cent w/v solution of sodium acetate and
1 ml of 0.1 M perchloric acid is equivalent to 0.01972 g of sufficient water to produce 50.0 ml and measure the optical
C9H11NO4. rotation at 20°. Separately dilute 5.0 ml of the filtrate to 200.0
Storage. Store protected from light and moisture. ml with 0.1 M hydrochloric acid, mix well and dilute 10.0 ml to
200.0 ml with 0.1 M hydrochloric acid. Measure the
280 nm (2.4.7). Calculate the content of C9H11NO4, in the filtrate
taking 142 as the specific absorbance at 280 nm and from the
Levodopa Tablets result so obtained calculate the specific optical rotation.
L-Dopa Tablets Related substances. Determine by thin-layer chromatography
Levodopa Tablets contain not less than 95.0 per cent and not (2.4.17), coating the plate with microcrystalline cellulose.
more than 105.0 per cent of the stated amount of levodopa, Mobile phase. A mixture of 50 volumes of 1-butanol, 25
C9H11NO4. volumes of glacial acetic acid and 25 volumes of water.
Identification
containing 0.1 g of Levodopa with 10 ml of a mixture of equal
A. Dissolve as completely as possible a quantity of the tablet volumes of anhydrous formic acid and methanol.
containing 0.5 g of Levodopa in 25 ml of 1 M hydrochloric Reference solution (a). Dilute 1 volume of the test solution to
acid and filter. Adjust the pH to 3 with 5 M ammonia, added 200 volumes with methanol.
dropwise with stirring, and allow to stand for several hours,
protected from light. Filter, wash the precipitate and dry it at Reference solution (b). A mixture of equal volumes of the test
105°. solution and a solution prepared by dissolving 30 mg of L-
tyrosine in 1 ml of anhydrous formic acid and diluting to 100
On the residue, determine by infrared absorption ml with methanol.
obtained with levodopa RS or with the reference spectrum of Apply to the plate 10 µl each of the test solution and reference
levodopa. solution (a) and 20 µl of reference solution (b) as bands. Allow
the mobile phase to rise 20 cm. Dry the plate in a current of
B. Determine by thin-layer chromatography (2.4.17), coating warm air, spray with a freshly prepared mixture of equal volumes
the plate with microcrystalline cellulose. of a 10 per cent w/v solution of ferric chloride and a 5 per cent
Mobile phase. A mixture of 50 volumes of 1-butanol, w/v solution of potassium ferricyanide and examine the plate
25 volumes of glacial acetic acid and 25 volumes of water. immediately. Any secondary band in the chromatogram
band in the chromatogram obtained with reference solution
containing 0.1 g of Levodopa with 10 ml of 1 M hydrochloric
acid and filter.
677
LEVOFLOXACIN HEMIHYDRATE IP 2007
with reference solution (b) shows a distinct band, at a higher Tests

Rf value than the principal band, which is more intense than
the band in the chromatogram obtained with reference solution D-ofloxacin. Determine by liquid chromatography (2.4.14).
(a). Test solution. Dissolve 10 mg of the substance under
Dissolution (2.5.2). examination in 50 ml of mobile phase.
Apparatus. No 2 Chromatographic system

Medium. 900 ml of 0.1 M hydrochloric acid – a stainless steel column 25 cm x 4 mm packed with
Withdraw a suitable volume of the medium and filter. Measure – mobile phase: dissolve 1.25 g of copper(II)sulphate
the absorbance of the resulting solution at the maximum at pentahydrate and 1.31 g of L-isolucine in 113 ml of
about 280 nm (2.4.7). Calculate the content of C9H11NO4 taking methanol and make 1000 ml with water,
141 as the specific absorbance at 280 nm. – flow rate. 1.8 ml per minute,
D. Not less than 75 per cent of the stated amount of C9H11NO4. – spectrophotometer set at 330 nm,
Inject the test solution. The relative retention times are about
Assay. Weigh and powder 20 tablets. Weigh accurately a 1.0 for levofloxacin and 1.2 for D-ofloxacin. The area of peak
quantity of the powder containing about 0.4 g of Levodopa, corresponds to D-ofloxacin in the chromatogram obtained with
dissolve as completely as possible in 10 ml of anhydrous the test solution is not more than 2.0 per cent of the area of the
formic acid, add 80 ml of anhydrous glacial acetic acid. Titrate principal peak.
with 0.1 M perchloric acid, using oracet blue B solution as
indicator. Carry out a blank titration. Heavy metals (2.3.13). 1.0 g complies with the limit test for
heavy metals, Method B ( 20 ppm).
C9H11NO4. Sulphated ash (2.3.18). Not more than 0.2 per cent, determined
on 1.0 g.
anhydrous acetic acid. Titrate with 0.1 M perchloric acid,
Levofloxacin Hemihydrate determining the end-point potentiometrically (2.4.25). Carry
H3C CH3
N O 1 ml 0.1 M perchloric acid is equivalent of 0.03614 g of
C18H20FN3O4.
N N
, 1/2 H2O Storage. Store protected from light.
F COOH
O
Levofloxacin Infusion
C18H20FN3O4, ½ H2O Mol. Wt. 370.4
Levofloxacin Infusion is a sterile solution of levofloxacin in
Levofloxacin Hemihydrate is (-)-(S)-9-fluoro-2,3-dihydro-3- water for injection.
methyl-10-(4-methylpiperazin-1-yl)-7-oxo-7H-pyrido[1,2,3-
de]-1,4-benzoxazine-6-carboxylic acid hemihydrate. Levofloxacin Infusion contain not less than 90.0 per cent and
Levofloxacin Hemihydrate contains not less than 98.5 per cent levofloxacin, C18H20FN3O4.
and not more than 101.0 per cent of C18H20FN3O4, calculated
on the anhydrous basis. Identification
Description. A yellowish white to yellow powder. In the Assay, the chromatogram obtained with the test solution
Identification reference solution.
Determine by infrared absorption spectrophotometry (2.4.6). Tests
Compare the spectrum with that obtained with levofloxacin
hemihydrate RS. pH (2.4.24). 3.8 to 5.8.
678
IP 2007 LEVOFLOXACIN TABLETS
Other tests. Complies with the tests stated under Parenteral Withdraw a suitable volume of the medium and filter. Measure
Preparation (Infusions). the absorbance of the filtered solution, suitably diluted with
Assay. Determine by liquid chromatography (2.4.14). the medium if necessary, at the maximum at about 326 nm
(2.4.7). Calculate the content of C18H20FN3O4 in the medium
NOTE — Use freshly prepared solutions and carry out the from the absorbance obtained from a solution of known
test protected from light. concentration of levofloxacin hemihydrate RS in the same
Test solution. Measure accurately a volume containing 50 mg medium.
of levofloxacin, dilute to 50.0 ml with 0.1 M hydrochloric acid. D. Not less than 70.0 per cent of the stated amount of
Dilute 5.0 ml of the solution to 25.0 ml with water. C18H20FN3O4.
Reference solution. A 0.1 per cent w/v solution of levofloxacin Related substances. Determine by liquid chromatography
hemihydrate RS in 0.1 M hydrochloric acid. Dilute 5.0 ml of (2.4.14).
the solution to 25.0 ml with a water.
NOTE — Use freshly prepared solutions and carry out the
Chromatographic system test protected from light.
octadecylsilane bonded to porous silica (5µm), Test solution. Weigh and powder 20 tablets. Weigh accurately
– mobile phase: a mixture of 85 volumes of buffer solution a quantity of powdered tablet containing 100 mg of
prepared by mixing 84 volumes of 0.05 M citric acid Levofloxacin, disperse in 100 ml of 0.1 M hydrochloric acid
monohydrate and 1 volume of 1 M ammonium acetate and filter. Dilute 5 ml of the solution to 10 ml with a water.
and 15 volumes of acetonitrile, Reference solutionn (a). A 0.1 per cent w/v solution of
– flow rate. 1 ml per minute, levofloxacin hemihydrate RS in 0.1 M hydrochloric acid.
– spectrophotometer set at 293 nm, Dilute 5 ml of the solution to 10 ml with a water.
– 10 µl loop injector.
Inject the reference solution. The test is not valid unless the 100 ml with water.
tailing factor is not more than 2.0 and column efficiency is not
less than 3000 theoretical plates and the relative standard Chromatographic system as described under Assay.
deviation is not more than 2.0 per cent. Inject reference solution (a). Test is not valid unless the column
Inject the test solution and the reference solution. effciency is not less than 2000 theoretical plates and the tailing
Calculate the content of C18H20FN3O4 in the infusion.
Levofloxacin Tablets (0.5 per cent) and the sum of areas of all the secondary peaks
is not more than the area of the peak in the chromatogram
Levofloxacin Tablets contain Levofloxacin hemihydrate. obtained with the reference solution (b) (1.0 per cent).
Levofloxacin Tablets contain not less than 90.0 per cent and Loss on drying (2.4.19). Not more than 6.0 per cent, determined
not more than 110.0 per cent of the stated amount of on 1 g by drying at 105° , under vaccume, for 3 hours.
levofloxacin, C18H20FN3O4.
Other tests. Comply with the tests stated under the Tablets.
Identification Assay. Determine by liquid chromatography (2.4.14).
In the Assay, the chromatogram obtained with the test solution NOTE — Use freshly prepared solutions and carry out the
corresponds to that in the chromatogram obtained with test protected from light.
reference solution.
Tests a quantity of powdered tablet containing 100 mg of levofloxacin,
disperse in 100.0 ml of 0.1 M hydrochloric acid and filter.
Dissolution (2.5.2). Dilute 5.0 ml of the solution to 50.0 ml with water.
Apparatus No. 2 Reference solutionn. A 0.1 per cent w/v solution of
Medium. 900 ml of 0.01 M hydrochloric acid. levofloxacin hemihydrate RS in 0.1 M hydrochloric acid.
Speed and time. 100 rpm for 45 minutes. Dilute 5 ml of the solution to 50 ml with a water.
679
LEVONORGESTREL IP 2007

octadecylsilane bonded to porous silica (5µm) (such as Specific optical rotation (2.4.22). –30.0° to –35.0°, determined
Cosmosil C18 MS II), in a 2.0 per cent w/v solution in chloroform.
– mobile phase: a mixture of 85 volume of buffer solution Related substances. Determine by thin-layer chromatography
prepared by dissolving 84 volumes of 0.05 M citric (2.4.17), coating the plate with silica gel G.
acid monohydrate and 1 volume of 1 M ammonium Mobile phase. A mixture of 80 volumes of dichloromethane
acetate, filter and 15 volumes of acetonitrile, and 20 volumes of ethyl acetate.
– spectrophotometer set at 293 nm, Test solution. Dissolve 0.2 g of the substance under
– 10 µl loop injector. examination in 10 ml of chloroform.
Inject the reference solution. The test is not valid unless the Reference solution (a). A 0.01 per cent w/v solution of the
tailing factor is not more than 2.0, column efficiency is not less substance under examination in chloroform.
than 3000 theoretical plates and the relative standard deviation Reference solution (b). A 0.004 per cent w/v solution of the
for replicate injections is not more than 2.0 per cent. substance under examination in chloroform
Inject the test solution and the reference solution. Apply to the plate 10 µl of each solution. After development,
dry the plate in air and spray with phosphomolybdic acid
Calculate the content of C18H20FN3O4 in the tablets.
Storage. Store protected from light and moisture. with the test solution is not more intense than the spot in the
more than two such spots are more intense than the spot in
the chromatogram obtained with reference solution (b).
Levonorgestrel Sulphated ash (2.3.18). Not more than 0.3 per cent.
H3C on 1.0 g by drying in an oven at 105°.
OH
C CH Assay. Weigh accurately about 0.1 g, dissolve in sufficient
H H ethanol (95 per cent) to produce 100.0 ml, dilute stepwise
with ethanol (95 per cent) to obtain a solution containing
H H 0.001 per cent w/v of Levonorgestrel and measure the
O absorbance of the resulting solution at the maximum at about
241 nm (2.4.7).
C21H28O2 Mol. Wt. 312.5 Calculate the content of C 21H28O2 from the absorbance
obtained with a 0.001 per cent w/v solution of levonorgestrel
Levonorgestrel is 13β-ethyl-17β-hydroxy-18,19-dinor-17α-
RS in ethanol (95 per cent).
pregn-4-en-20-yn-3-one.
Storage. Store protected from light and moisture, at a
Levonorgestrel contains not less than 98.0 per cent and not
more than 102.0 per cent of C21H28O2, calculated on the dried
basis.
odourless or almost odourless. Levonorgestrel And Ethinyloestradiol
Identification
Tablets
Levonorgestrel and Ethinyloestradiol Tablets contain not less
Compare the spectrum with that obtained with levonorgestrel
stated amounts of levonorgestrel, C 21 H 28 O 2 , and
RS or with the reference spectrum of levonorgestrel.
ethinyloestradiol, C20H24O2. The tablets may be film-coated.
0.001 per cent w/v solution in methanol shows an absorption Identification
C. Melts at about 237° (2.4.21). plate with silica gel G.
680
IP 2007 LIGNOCAINE HYDROCHLORIDE
Mobile phase. A mixture of 96 volumes of dichloromethane Reference solution. Transfer 15.0 ml of a solution of
and 4 volumes of ethanol (95 per cent). levonorgestrel RS in the mobile phase and 3.0 ml of a solution
Test solution. Powder 20 tablets finely, triturate with 20 ml of of ethinyloestradiol RS in the mobile phase, each solution
dichloromethane, allow the solids to sediment and use the having a concentration of about 0.1 mg per ml, into a 100-ml
clear supernatant liquid. volumetric flask. Add 10 ml of a 0.0025 per cent w/v solution of
diphenyl (internal standard) in the mobile phase. Dilute to
Reference solution. A solution containing 0.06 per cent w/v of volume with the mobile phase and mix. Each ml of this reference
levonorgestrel RS and 0.006 per cent w/v of ethinyloestradiol solution has a known concentration of about 15 µg and 3 µg
RS in water. of levonorgestrel and ethinyl oestradiol per ml respectively.
Apply to the plate 40 µl of each solution. After development, Chromatographic system
dry the plate in air, spray with ethanolic sulphuric acid
(80 per cent v/v), heat at 110° for 10 minutes and examine in
ultraviolet light at 365 nm. The principal spots in the
chromatogram obtained with the test solution correspond to
15 volumes of methanol and 45 volumes of water,
the spots for levonorgestrel (red fluorescence) and
– flow rate. 1 to 1.5 ml per minute,
ethinyloestradiol (orange-yellow fluorescence) in the
Tests Inject the reference solution and record the peak response.
Uniformity of content. Comply with the test stated under The test is not valid unless the resolution between the two
Tablets. major peaks is not less than 2.5, and the relative standard
deviation for replicate injections is not more than 2.0.
Carry out the procedure described under Assay but using the
following solutions. Inject separately the test solution and the reference solution
and measure the responses for the major peaks. The relative
Test solution. Add 2.0 ml of methanol (70 per cent) and 2.0 ml
retention times are about 0.7 for ethinyloestradiol and 1.0 for
of a 0.00002 per cent w/v solution of diphenyl in methanol
levonorgestrel.
(70 per cent) (internal standard) to one tablet, shake for
20 minutes, centrifuge, filter the supernatant liquid through a Calculate the contents of C21H28O2 and C20H24O2 in the tablets.
membrane filter with a pore size of not more than 0.2 µm and
use the filtrate.
Reference solution (a). Weigh accurately a quantity of
norgestrel RS containing 100 times the stated amount of
Levonorgestrel per tablet, dissolve in sufficient methanol
(70 per cent) to produce 200.0 ml. Take 2.0 ml of this solution,
Lignocaine Hydrochloride
add 2.0 ml of the internal standard solution and use the resulting Lidocaine Hydrochloride
solution.
Reference solution (b). Weigh accurately a quantity of
CH3 H
ethinyloestradiol RS containing 100 times the stated amount
of Ethinyloestradiol per tablet, dissolve in sufficient methanol N
N CH3 , HCl, H2O
(70 per cent) to produce 200.0 ml. Take 2.0 ml of the solution, O
add 2.0 ml of the internal standard solution and use the resulting CH3 CH3
solution.
Other tests. Comply with the tests stated under Tablets. C14H22N2O,HCl,H2O Mol. Wt. 288.8
Assay. Determine by liquid chromatography (2.4.14). Lignocaine Hydrochloride is 2-diethylaminoaceto-2’,6’-
Test solution. Weigh and finely powder 20 tablets. To a quantity xylidide hydrochloride monohydrate.
of the powdered tablets containing about 3 mg of
Lignocaine Hydrochloride contains not less than 99.0 per cent
Levonorgestrel in a 200-ml volumetric flask, add the mobile
and not more than 101.0 per cent of C14H22N2O,HCl, calculated
phase and 10 ml of a 0.0025 per cent w/v solution of diphenyl
(internal standard) in the mobile phase. Shake by mechanical
means for 20 minutes and dilute to volume with the mobile Description. A white, crystalline powder; odourless or
phase. Centrifuge and use the clear supernatant liquid. practically odourless.
681
LIGNOCAINE AND ADRENALINE INJECTION IP 2007
Identification solution. Titrate with 0.1 M perchloric acid, using crystal

violet solution as indicator. Carry out a blank titration.
Tests B, C, D and E may be omitted if test A is carried out. 1 ml of 0.1 M perchloric acid is equivalent to 0.02708 g of
C14H22N2O,HCl.
Compare the spectrum with that obtained with lignocaine Storage. Store protected from moisture.
hydrochloride RS or with the reference spectrum of lignocaine
hydrochloride.
B. To 10 ml of a 2.5 per cent w/v solution add sodium hydroxide
solution till alkaline and filter. Wash the residue with water, Lignocaine and Adrenaline Injection
dissolve half of the residue in 1 ml of ethanol (95 per cent)
Lidocaine Hydrochloride and Adrenaline Bitartrate
and add 0.5 ml of a 10 per cent w/v solution of cobalt chloride;
a bluish-green precipitate is produced.
Injection; Lidocaine and Adrenaline Injection
C. To 5 mg add 0.5 ml of fuming nitric acid, evaporate to Lignocaine and Adrenaline Injection is a sterile solution of
dryness on a water-bath, cool, dissolve the residue in 5 ml of Lignocaine Hydrochloride and Adrenaline Bitartrate in Water
acetone and add 1 ml of 0.1 M ethanolic potassium for Injections.
hydroxide; a green colour is produced. Lignocaine and Adrenaline Injection contains not less than
D. Dissolve 0.2 g in 10 ml of water and add 10 ml of picric acid 95.0 per cent and not more than 105.0 per cent of the stated
solution. The precipitate, after washing with water and drying, amount of lignocaine hydrochloride, C14H22N2O,HCl,H2O and
melts at about 229° (2.4.21). not less than 87.5 per cent and not more than 112.5 per cent of
the stated amount of adrenaline, C9H13NO3.
E. Gives reaction A of chlorides (2.3.1).
Description. A clear colourless solution.
Tests
Identification
dioxide-free water is clear (2.4.1), and colourless (2.4.1). A. To 5 ml add 1 ml of hydrochloric acid, cool to 0°, add 5 ml
of a 1 per cent w/v solution of sodium nitrite and pour the
pH (2.4.24). 4.0 to 5.5, determined in a 0.5 per cent w/v solution. mixture into 2 ml of 2-naphthol solution containing 1 g of
Heavy metals (2.3.13). 2.0 g complies with the limit test for sodium acetate; no red colour is produced.
heavy metals, Method A (10 ppm). B. To 10 ml add 4 ml of disodium hydrogen phosphate solution
Sulphates. Dissolve 0.2 g in 20 ml of water, add 2 ml of 3 M and sufficient 0.1 M iodine to produce a distinct brown colour.
hydrochloric acid, mix and divide into two parts. To one part Add 0.01 M sodium thiosulphate to remove the excess of
add 1 ml of barium chloride solution; no more opalescence is iodine; a pink colour is produced.
produced than in the remaining portion of the solution to C. To 3 ml add 3 ml of water and 6 ml of picric acid solution,
which nothing has been added. shake gently and allow to stand until the precipitate becomes
2,6-Dimethylaniline. To 2 ml of a 2.5 per cent w/v solution in crystalline; the precipitate, after washing with water and
methanol (solution A), add 1 ml of a 1 per cent w/v solution of drying at 105°, melts at about 229° (2.4.21).
4-dimethylaminobenzaldehyde in methanol and 2 ml of
glacial acetic acid and allow to stand for 10 minutes at room Tests
temperature. Any yellow colour produced is more intense than
pH (2.4.24). 3.0 to 4.5.
that obtained by repeating the test using 2 ml of methanol in
place of solution A and less intense than the colour produced Other tests. Complies with the tests stated under Parenteral
using a mixture of 1 ml of a solution of 2,6-dimethylaniline in Preparations (Injections).
methanol containing 5 µg per ml and 1 ml of methanol in place
Assay. For lignocaine hydrochloride — Make an accurately
of solution A (100 ppm).
measured volume containing 0.1 g of Lignocaine Hydrochloride
Sulphated ash (2.3.18). Not more than 0.1 per cent. alkaline with 2 M sodium hydroxide and extract with three
quantities, each of 20 ml, of chloroform, washing each extract
with the same 10 ml of water. Filter the washed extracts through
Assay. Weigh accurately about 0.5 g, dissolve in 30 ml of a filter paper moistened with chloroform, wash the filter with
anhydrous glacial acetic acid, add 6 ml of mercuric acetate 10 ml of chloroform, combine the washings with the
682
IP 2007 LIGNOCAINE AND DEXTROSE INJECTION
filtrate.Titrate with 0.02 M perchloric acid, using crystal violet B. To a volume containing 0.1 g of Dextrose add 10 ml of water
solution as indicator. and 3 ml of potassium cupri-tartrate solution and heat; a red
1 ml of 0.02 M perchloric acid is equivalent to 0.005776 g of precipitate is produced.
C14H22N2O,HCl,H2O. Tests
For adrenaline — To an accurately measured volume pH (2.4.24). 3.0 to 7.0.
containing 0.1 mg of adrenaline add 20 mg of sodium
metabisulphite, 0.1 ml of ferrous sulphate-citrate solution, 1 Other tests. Complies with the tests described under Parenteral
ml of glycine buffer solution and mix. Allow to stand for 10 Preparations (Injections).
minutes, extract with 10 ml of ether, allow to separate, reject Assay. For lignocaine hydrochloride - Make an accurately
the ether and measure the absorbance of a 4-cm layer of the measured volume containing about 0.1 g of Lignocaine
solution at about 540 nm (2.4.7). Calculate the content of Hydrochloride alkaline with 2 M sodium hydroxide and extract
adrenaline, C9H13NO3, from a reference curve prepared by with three quantities, each of 20 ml, of chloroform, washing
treating suitable aliquots of a solution of adrenaline bitartrate each extract with the same 10 ml of water. Filter the washed
RS in the same manner. extracts through a filter paper moistened with chloroform,
1 mg of adrenaline bitartrate is equivalent to 0.0005497 g of wash the filter with 10 ml of chloroform, combine the washings
C9H13NO3. with the filtrate. Titrate with 0.02 M perchloric acid, using
crystal violet solution as indicator.
Labelling. The label states the strength of Adrenaline Bitartrate C14H22N2O,HCl,H2O.
in terms of the equivalent amount of adrenaline.
For dextrose — To an accurately measured volume containing
2 g to 5 g of Dextrose add sufficient water to produce 100.0 ml.
Mix well, allow to stand for 30 minutes and determine the
optical rotation in a 2-dm tube (2.4.22). The observed rotation
Lignocaine and Dextrose Injection in degrees multiplied by 1.0425 represents the weight, in g, of
dextrose, C6H12O6,H2O in the volume taken for assay.
Lignocaine Hydrochloride and Dextrose Injection;
Lidocaine Hydrochloride and Dextrose Injection; Storage. Store in single dose containers in a cool place.
Lidocaine and Dextrose Injection
Lignocaine and Dextrose Injection is a sterile solution of
Lignocaine Hydrochloride and Dextrose in Water for Injections. Lignocaine Gel
Lignocaine and Dextrose Injection contains not less than 95.0 Lignocaine Hydrochloride Gel; Lidocaine Hydrochloride
per cent and not more than 105.0 per cent of the stated amounts Gel
of lignocaine hydrochloride, C14H22N2O,HCl,H2O, and dextrose,
Lignocaine Gel is a sterile solution of Lignocaine Hydrochloride
C6H12O6,H2O.
in a suitable water-miscible base. It may contain suitable
Description. A clear colourless or slightly yellow solution. antioxidants, stabilisers and antimicrobial preservatives.
Lignocaine Gel contains not less than 95.0 per cent and not
Identification
A. To a volume containing about 0.5 g of Lignocaine lignocaine hydrochloride, C14H22N2O,HCl.
Hydrochloride in a separator add 2 ml of 2 M sodium hydroxide
and extract with four quantities, each of 15 ml, of chloroform. Identification
Combine the chloroform extracts and evaporate the solution To a quantity of the gel containing 80 mg of anhydrous
to dryness with the aid of a current of air. Dissolve the residue lignocaine hydrochloride add 4 ml of hydrochloric acid and
in 2 ml of hexane, evaporate with the aid of warm air and dry heat on a water-bath for 10 minutes. Allow to cool, transfer to
the residue over silica gel for 24 hours at a pressure not a separating funnel with the aid of 20 ml of water, add 5 M
exceeding 0.7 kPa. sodium hydroxide until precipitation is complete and extract
On the residue, determine by infrared absorption with two quantities, each of 20 ml, of chloroform. Filter the
spectrophotometry (2.4.6). Compare the spectrum with that chloroform extracts through anhydrous sodium sulphate and
obtained with lignocaine hydrochloride RS treated in the evaporate the filtrate to dryness on a water-bath using a stream
same manner. of nitrogen. The residue complies with the following tests.
683
LIGNOCAINE INJECTION IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). Lignocaine Injection

Compare the spectrum with that obtained with lignocaine
hydrochloride RS or with the reference spectrum of lignocaine Lignocaine Hydrochloride Injection; Lidocaine
hydrochloride. Hydrochloride Injection; Lidocaine Injection
B. Dissolve 20 mg in 1 ml of ethanol (95 per cent), add 0.5 ml Lignocaine Injection is a sterile solution of Lignocaine
of a 10 per cent w/v solution of cobalt chloride and 0.5 ml of Hydrochloride in Water for Injections.
5 M sodium hydroxide and shake for 2 minutes; a bluish green
precipitate is produced. Lignocaine Injection contains not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of lignocaine
C. Dissolve 40 mg in 5 ml of 1 per cent w/v solution of cetrimide, hydrochloride, C14H22N2O,HCl,H2O.
add 1 ml of 5 M sodium hydroxide and 1 ml of bromine water;
a yellow colour is produced. Description. A clear colourless solution.
pH (2.4.24). 6.0 to 7.0. A. To a volume containing 0.1 g of Lignocaine Hydrochloride

add sufficient sodium hydroxide solution to make alkaline,
2,6-Dimethylaniline. Mix a quantity of the gel containing filter, wash the residue with water, dissolve in 1 ml of ethanol
15 mg of anhydrous lignocaine hydrochloride with sufficient (95 per cent), add 0.5 ml of a 10 per cent w/v solution of
water to produce 3 ml, using a rotary mixer. To 2 ml of the cobalt chloride and shake for 2 minutes; a bluish-green
resulting solution, add 1 ml of a freshly prepared 1 per cent w/ precipitate is formed.
v solution of 4-dimethylaminobenzaldehyde in methanol. Mix
thoroughly using a rotary mixer. Add 2 ml of glacial acetic B. To a volume containing 0.1 g of Lignocaine Hydrochloride
acid and allow to stand for 10 minutes. Any yellow colour add 10 ml of picric acid solution; the precipitate, after washing
produced is not more intense than that obtained by using a with water and drying at 105°, melts at about 229° (2.4.21).
mixture of 2 ml of a solution of 2,6-dimethylaniline in methanol C. Gives the reactions of chlorides (2.3.1).
containing 2 µg per ml in place of the solution of the gel
(20 ppm). Tests
Sterility (2.2.11). Complies with the test for sterility. pH (2.4.24). 5.0 to 7.0.
Other tests. Complies with the tests stated under Gels. 2,6-Dimethylaniline. To a volume containing 25 mg of
Assay. Weigh accurately a quantity containing about 10 mg Lignocaine Hydrochloride add water if necessary to produce
of anhydrous lignocaine hydrochloride and disperse in 20 ml 10 ml, add 2 M sodium hydroxide until the solution is just
of water. Add 5 ml of acetate buffer pH 2.8, 120 ml of chloroform alkaline and extract with three quantities, each of 5 ml, of
and 5 ml of dimethyl yellow-oracet blue B solution and titrate chloroform. Dry the combined chloroform extracts over
with 0.005 M dioctyl sodium sulphosuccinate swirling anhydrous sodium sulphate, filter, wash with a further 5 ml of
vigorously, until near the end-point, then add the titrant chloroform and evaporate the filtrate to dryness at a pressure
dropwise and, after each addition, swirl vigorously, allow to of 2 kPa. Dissolve the residue in 2 ml of methanol, add 1 ml of
separate and swirl gently for 5 seconds. The end-point is a 1 per cent w/v solution of 4-dimethylamino-benzaldehyde
indicated when the colour of the chloroform layer changes in methanol and 2 ml of glacial acetic acid and allow to stand
from green to pinkish-grey. Carry out a blank titration. at room temperature for 10 minutes. Any yellow colour
produced is not more intense than the colour produced by
1 ml of 0.005 M dioctyl sodium sulphosuccinate is equivalent repeating the operation using 10 ml of a solution in water
to 0.001354 g of C14H22N2O,HCl. containing 1 µg per ml of 2,6-dimethylaniline in place of the
Determine the weight per ml of the gel (2.4.29), and calculate preparation under examination (400 ppm).
the percentage of C14H22N2O,HCl, weight in volume. Other tests. Complies with the tests stated under Parenteral
Storage. Store in suitable tamper-proof containers holding Preparations (Injections).
sufficient of the gel for use on one occasion, and at a Assay. Make an accurately measured volume containing about
temperature not exceeding 30°. The gel should not be frozen. 0.1 g of Lignocaine Hydrochloride alkaline with 2 M sodium
Labelling. The label states (1) that the contents are sterile; (2) hydroxide and extract with three quantities, each of 20 ml, of
the strength in terms of the equivalent amount of anhydrous chloroform, washing each extract with the same 10 ml of water.
lignocaine hydrochloride; (3) that any of the gel not used in a Filter the washed extracts through a filter paper moistened
single application should be discarded. with chloroform, wash the filter with 10 ml of chloroform,
684
IP 2007 LINCOMYCIN HYDROCHLORIDE
combine the washings with the filtrate. Titrate with 0.02 M Reference solution (b). A solution containing 0.1 per cent
perchloric acid, using crystal violet solution as indicator. w/v each of lincomycin hydrochloride RS and clindamycin
1 ml of 0.02 M perchloric acid is equivalent to 0.005776 g of hydrochloride RS in methanol.
C14H22N2O,HCl,H2O. Apply to the plate 5 µl of each solution. After development,
dry the plate in air and spray with a 0.1 per cent w/v solution
of potassium permanganate. The principal spot in the
Lincomycin Hydrochloride that in the chromatogram obtained with reference solution (a).
reference solution (b) shows two clearly separated spots.
CH3 C. Dissolve 10 mg in 2 ml of 2 M hydrochloric acid and heat in
CH3
H3C N a water-bath for 3 minutes. Add 3 ml of a 10.5 per cent w/v
HO H
solution of anhydrous sodium carbonate and 1 ml of a 2 per
N H
H cent w/v solution of sodium nitroprusside; a violet-red colour
O
is produced.
HO O , HCl, H2O
OH D. A 1 per cent w/v solution gives reaction A of chlorides
SCH3 (2.3.1).
OH
Tests
C18H34N2O6S,HCl,H2O Mol. Wt. 461.0 Appearance of solution. A 10.0 per cent w/v solution in carbon
Lincomycin Hydrochloride consists mainly of methyl 6- dioxide-free water is clear (2.4.1), and not more intensely
amino-6,8-dideoxy-N-[(2S,4R)-1-methyl-4-propylprolyl]-1- coloured than reference solution YS6 (2.4.1).
thio-D-erythro-α-D-galacto-octopyranoside hydrochloride
monohydrate, an antimicrobial substance produced by
Streptomyces lincolnensis var. lincolnensis or by any other Specific optical rotation (2.4.22). +135° to +150°, determined
means. in a 4.0 per cent w/v solution.
Lincomycin Hydrochloride contains not less than 82.5 per Lincomycin B. In the Assay, the chromatogram obtained with
cent and not more than 93.0 per cent of C18H34N2O6S, calculated reference solution (a) shows a peak derived from lincomycin
on the anhydrous basis. B which is eluted just before lincomycin. The area of the peak
derived from lincomycin B is not more than 5 per cent of the
Description. A white or almost white, crystalline powder. area of the peak derived from lincomycin.
Identification Heavy metals (2.3.13). 2.0 g complies with the limit test for
B and C may be omitted if tests A and D are carried out. Sulphated ash (2.3.18). Not more than 0.5 per cent.
Compare the spectrum with that obtained with lincomycin Assay. Determine by gas chromatography (2.4.13).
hydrochloride RS or with the reference spectrum of lincomycin Test solution. Add 10.0 ml of a 0.8 per cent w/w solution of
hydrochloride. dotriacontane (internal standard) in chloroform to 0.1 g of
B. Determine by thin-layer chromatography (2.4.17), coating lincomycin hydrochloride RS, dilute to 100.0 ml with a 2 per
the plate with silica gel G. cent w/v solution of imidazole in chloroform and shake to
dissolve. Place 4.0 ml of the resulting solution in a 15-ml glass-
Mobile phase. The upper layer obtained by shaking a mixture
stoppered centrifuge tube, add 1.0 ml of a mixture of 99 volumes
of 45 volumes of ethyl acetate, 40 volumes of a 15 per cent
of N,O-bis (trimethylsilyl)-acetamide and 1 volume of
w/v solution of ammonium acetate previously adjusted to pH
trimethylchlorosilane and mix gently. Loosen the glass
9.6 with 10 M ammonia and 20 volumes of 2-propanol.
stopper and heat at 65° for 30 minutes.
solution but omitting the internal standard and using 0.1 g of
Reference solution (a). A 0.1 per cent w/v solution of the substance under examination in place of lincomycin
lincomycin hydrochloride RS in methanol. hydrochloride RS.
685
LINCOMYCIN CAPSULES IP 2007
Reference solution (b). Prepare in the same manner as the test hydrochloride RS relative to that of the internal standard in
solution but using 0.1 g of the substance under examination the test solution.
in place of lincomycin hydrochloride RS.
Tests
– a glass column 1.5 m x 3 mm, packed with acid-washed, Lincomycin B. Examine reference solution (b) as described
silanised diatomaceous support impregnated with 3 per under the Assay but increase the sensitivity by 8 to 10 times
cent w/w of phenyl methyl silicone fluid (50 per cent while recording the peak derived from lincomycin B, which is
phenyl), eluted immediately before that derived from lincomycin. The
– temperature: area of the peak derived from lincomycin B, when corrected
column 260°, for the sensitivity factor, is not more than 5 per cent of the area
inlet port and detector at 260° to 290°, of the peak derived from lincomycin.
– flow rate. 45 ml per minute using helium as carrier gas.
Inject 1 µl of the test solution, reference solution (a) and 0.3 g of the contents of the capsules.
Calculate the content of C18H34N2O6S.
Lincomycin Hydrochloride intended for use in the
manufacture of parenteral preparations without a further Test solution. Add 10.0 ml of a 0.8 per cent w/w solution of
appropriate procedure for the removal of bacterial dotriacontane (internal standard) in chloroform to 0.1 g of
endotoxins complies with the following additional lincomycin hydrochloride RS, dilute to 100.0 ml with a 2 per
requirement. cent w/v solution of imidazole in chloroform and shake to
dissolve. Place 4.0 ml of the resulting solution in a 15-ml glass-
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit stoppered centrifuge tube, add 1.0 ml of a mixture of 99 volumes
per mg. of N,O-bis(trimethylsilyl)-acetamide and 1 volume of
Lincomycin Hydrochloride intended for use in the trimethylchlorosilane and mix gently. Loosen the glass
manufacture of parenteral preparations without a further stopper and heat at 65° for 30 minutes.
solution but omitting the internal standard and using a quantity
Sterility (2.2.11). Complies with the test for sterility. of the mixed contents of 20 capsules containing about 90 mg
of lincomycin in place of lincomycin hydrochloride RS.
exceeding 30°. If the contents are sterile, the container should Reference solution (b). Prepare in the same manner as the test
be sterile, tamper-evident and sealed so as to exclude micro- solution but using a quantity of the mixed contents of 20
organisms. capsules containing about 90 mg of lincomycin in place of
lincomycin hydrochloride RS.
Labelling. The label states whether or not the material is
intended for use in the manufacture of parenteral preparations. Chromatographic system
silanised diatomaceous support impregnated with 3 per
cent w/w of phenyl methyl silicone fluid (50 per cent
Lincomycin Capsules phenyl),
– temperature:
Lincomycin Hydrochloride Capsules column 260°,
Lincomycin Capsules contain Lincomycin Hydrochloride inlet port and detector at 260° to 290°,
equivalent to not less than 90.0 per cent and not more than – flow rate. 45 ml per minute using helium as carrier gas.
110 per cent of the stated amount of lincomycin, C18H34N2O6S. Inject 1 µl of the test solution, reference solution (a) and
Identification
Calculate the content of C18H34N2O6S in the capsules.
In the Assay, the retention time of the principal peak derived
from lincomycin hydrochloride relative to that of the internal
standard in reference solution (b) is the same as the retention Labelling. The label states the strength in terms of the
time of the principal peak derived from lincomycin equivalent amount of lincomycin.
686
IP 2007 ABSORBENT LINT
Lindane Apply separately to the plate 1 µl of each solution. Allow the

mobile phase to rise 12 cm. Dry the plate in a current of dry air
Gamma Benzene Hexachloride and irradiate with ultraviolet light at 254 nm for 15 minutes.
Spray with a 0.6 per cent w/v solution of dicarboxidine
Cl Cl hydrochloride in ethanol (90 per cent) and examine the spots
in daylight. Any secondary spot in the chromatogram obtained
Cl Cl with the test solution is not more intense than the spot in the
chromatogram obtained with reference solution (b). The test
Cl Cl
C6H6Cl6 Mol.Wt.290.8 solution (c) shows two clearly separated spots.
Lindane is 1α,2α,3β,4α,5,6β-hexachlorocyclohexane. Chlorides (2.3.12). To 0.75 g, finely powdered, add 15 ml of

water and boil for 1 minute. Allow to cool, shaking frequently,
Lindane contains not less 99.0 per cent and not more than and filter. To 10 ml of the filtrate add 3 ml of water and 2 ml of
100.5 per cent of C6H6Cl6. ethanol (95 per cent). The solution complies with the limit
Description. A white or almost white crystalline powder. test for chlorides (l00 ppm).
Identification
Assay. Weigh accurately about 0.2 g, add 10 ml of ethanol (95
A. Determine by infrared absorption spectrophotometry (2.4.6). per cent) and warm on a water-bath until dissolved. Cool, add
Compare the spectrum with that obtained with lindane RS. 20 ml of O.5 M ethanolic potassium hydroxide and allow to
B. In the test for Related substances, the principal spot in the stand for 10 minutes, swirling frequently. Add 50 ml of water,
chromatogram obtained with test solution (b) corresponds to 20 ml of 2 M nitric acid, 25 ml of 0.1 M silver nitrate and 5 ml
that in the chromatogram obtained with reference solution (a). of ferric ammonium sulphate solution. Titrate with 0.1 M
ammonium thiocyanate until a reddish-yellow colour is
C. Dissolve about 5 mg in 4 ml of ethanol (95 per cent). Add obtained. Repeat the procedure without the substance under
1 ml of 0.5 M ethanolic potassium hydroxide and allow to examination. The difference between the titrations represents
stand for 10 minutes. The solution gives reaction A of chlorides the amount of silver nitrate required.
(2.3.1).
1 ml of 0.1 M silver nitrate is equivalent to 0 009694 g of
D. Melts at 112º to 115º (2.4.21). C6H6Cl6.
Appearance of solution. A 5.0 per cent w/v solution in acetone

is clear (2.4.1), and not more intensely coloured than reference
solution YS7 (2.4.1).
Absorbent Lint
(2.4.17) coating the plate with silica gel G. Lint; Cotton Lint; Unmedicated Lint
Mobile phase. A mixture of 90 volumes of cyclohexane and 10 Absorbent Lint is a cotton cloth of plain weave, on one side
volumes of chloroform. of which a nep has been raised from either warp or weft yarns.
Test solution (a). Dissolve 1 g of the substance under It absorbs water readily but its absorbency may be
examination in 10 ml of chloroform. considerably reduced by medication, the absorbency of the
product depending upon the medicament incorporated.
chloroform. Description. Cotton cloth of plain weave, reasonably free from
weaving defects, readily tearable in both directions and
Reference solution (a). A 1 per cent w/v solution of lindane bleached to a good white having on one side a nep raised
RS in chloroform. from either the warp or weft yarns and reasonably free from
Reference solution (b). Dilute 1 ml of test solution (b) to 10 ml neps; it is clean and reasonably free from leaf, shell and other
with chloroform. foreign substances. It is made of yarn that is reasonably free
from slubs, snarls and other defects.
Reference solution (c). Dissolve 10 mg of α-hexachloro-
cyclohexane RS in sufficient of the test solution (a) to produce Absorbent Lint has not less than 98.0 per cent of the
5 ml. dimensions stated on the label.
687
LISINOPRIL IP 2007
Tests Related substances. Determine by liquid chromatography

(2.4.14).
Threads per cm. Warp not less than 16 and weft not less than
10. Test solution. Dissolve 20 mg of the substance under
examination in 10 ml of mobile phase A
Weight per unit area. 25 g has a superficial area of 1350 to
1370 sq. cm. Reference solution (a). A 0.2 per cent w/v solution of
lisinopril RS in mobile phase A.
Absorbency. A piece 10 cm square, placed lightly by means of
forceps, unraised side downwards, on the surface of water at Reference solution (b). Dilute 1 ml of reference solution (a) to
20°, becomes saturated within 10 seconds. 100 ml with mobile phase A.
Fluorescence. Not more than a few points of fluorescence are Chromatographic system
visible under screened ultraviolet light. – a stainless steel column 25 cm x 4.6 mm packed with
octylsilane bonded to porous silica (5 µm) (such as
Storage. Store protected from moisture in well-closed packages Symmetry C-8),
in a dry place, free from dust. – column temperature 50º,
Labelling. The label states the dimensions viz. the length and – mobile phase: A. a mixture of 30 volumes of acetonitrile
width in cm. and 970 volumes of 0.02 M sodium dihydrogen
phosphate, adjust the pH to 5.0 with a 5 per cent w/v
solution of sodium hydroxide and filter.
B. a mixture of 200 volumes of acetonitrile
Lisinopril and 800 volumes of 0.02 M sodium dihydrogen
phosphate, adjust the pH to 5.0 with 5 per cent w/v
solution of sodium hydroxide and filter.
NH2
below,
HOOC – spectrophotometer set at 210 nm,
N , 2H2O – a 20 µl loop injector.
N
H O COOH (min.) ( per cent) ( per cent)
0 100 0
C21H31N3O5, 2H2O Mol. Wt. 441.5 35 70 30
2
45 70 30
Lisinopril is (S)-1-[N -(1-carboxy-3-phenylpropyl)-L-lysyl]-L-
55 0 100
proline dihydrate.
65 100 0
Lisinopril contains not less than 98.0 per cent and not more 75 100 0
than 102.0 per cent of C21H31N3O5, calculated on the anhydrous
Inject reference solution (b). Test is not valid unless the tailing
basis.
Description. A white crystalline powder.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). in the chromatogram obtained with reference solution (b) (0.5
Compare the spectrum with that obtained with lisinopril RS per cent) and the sum of areas of all the secondary peaks is
or with the reference spectrum of lisinopril. not more than the area of the peak in the chromatogram
B. In the Assay, the chromatogram obtained with the test
solution corresponds to the chromatogram obtained with the Heavy metals (2.3.13). 1.0 g complies with the limit test for
reference solution. heavy metals, Method B ( 20 ppm).
Tests Sulphated ash (2.3.18). Not more than 0.1 per cent.
Water (2.3.43). 8.0 to 9.5, determined on 0.1 g.
Specific optical rotation (2.4.22). -115.0º to -123.0º, determined
on 1.0 per cent w/v solution in 0.25 M zinc acetate at 405 nm. Assay. Determine by liquid chromatography (2.4.14).
688
IP 2007 LISINOPRIL TABLETS
Test solution. Dissolve 30 mg of the substance under Chromatographic system

examination in 100 ml of water and filter. – a stainless steel column 25 cm x 4.6 mm packed with
Reference solution. A 0.03 per cent w/v solution of lisinopril octylsilane bonded to porous silica (5 µm) (such as
RS in water. Symmetry C8),
Chromatographic system – mobile phase: dissolve 1.0 g of hexane sulphonic acid
– a stainless steel column 25 cm x 4.6 mm packed with sodium salt in 800 volumes of phosphate solution
dimethyloctylsilane (C8 alkyl chain) fully endcapped prepared by dissolving 4.1 g of monobasic potassium
stationary phase (5 µm) (such as Hypersil MOS), phosphate in 900 ml water, adjust pH to 2.0 with
– column temperature 50º, orthophosphoric acid, dilute to 1000 ml with water and
– mobile phase: a mixture of 96 volumes of buffer pH 5.0 200 volumes of acetonitrile, mix, filter and degas.
prepared by dissolving 2.76 g of monobasic sodium – flow rate. 1.5 ml per minute,
phosphate in 1000 ml of water. adjust the pH to 5.0 with – spectrophotometer set at 215 nm,
1 M sodium hydroxide and 4 volumes of acetonitrile, – a 50 µl loop injector.
column efficiency is not less than 850 theoretical plates, the
Inject the reference solution. The test is not valid unless the deviation for replicate injections is not more than 2.0 per cent.
column efficiency is not less than 180 theoretical plates. The
deviation for replicate injections is not more than 2.0 per cent. D. Not less than 75 per cent of the stated amount of
C21H31N3O5.
(2.4.14).
Storage. Store protected from moisture, at a temperature not Test solution. Weigh accurately a powdered tablet containing
exceeding 25°. 100 mg of Lisinopril to a 50-ml volumetric flask. Add about 25
ml of mobile phase A and sonicate for 10 minutes with
intermittent shaking. Make up the volume with the mobile
Lisinopril Tablets phase A and filter.
Lisinopril Tablets contain not less than 90.0 per cent and not
lisinopril RS in the mobile phase A.
more than 110.0 per cent of the stated amount of lisinopril,
C21H31N3O5. Reference solution (b). Dilute 1 ml of reference solution (a) to
In the Assay, the principal peak in the chromatogram obtained – a stainless steel column 25 cm x 4.6 mm packed with
with the test solution corresponds to the peak in the packed with dimethyloctylsilane (C8 alkyl chain) fully
chromatogram obtained with the reference solution. endcapped stationary phase (5 µm) (such as Hypersil
MOS/ Symmetry C8),
Tests – column temperature 50º,
– mbile phase: A. a mixture of 3 volumes of acetonitrile
Dissolution (2.5.2). and 97 volumes of 0.02 M sodium dihydrogen
Apparatus. No 1 phosphate, adjust the pH to 5.0 with 5 per cent w/v
Medium. 900 ml of 0.1 M hydrochloric acid. solution of sodium hydroxide and filter,
Speed and time. 50 rpm for 30 minutes. B. a mixture of 20 volumes of acetonitrile
and 80 volumes of 0.02 M sodium dihydrogen
Withdraw a suitable volume of the medium and filter. phosphate, adjust the pH to 5.0 with 5 per cent w/v
Determine by liquid chromatography (2.4.14). solution of sodium hydroxide and filter,
Test solution. The filtrate obtained as given above.
below,
Reference solution. A 0.001 per cent w/v solution of lisinopril – flow rate. 1.8 ml per minute,
RS in dissolution medium. – spectrophotometer set at 210 nm,
689
LITHIUM CARBONATE IP 2007
– a 20 µl loop injector. Storage. Store protect from moisture, at a temperature between

Time Mobile phase A Mobile phase B 20° to 25°.
(min.) ( per cent) ( per cent)
0 100 0
35 70 30
Lithium Carbonate
45 70 30
Li2CO3 Mol.Wt.73.9
50 100 0
60 100 0 Lithium Carbonate contains not less than 98.5 per cent and
not more than 100.5 per cent of Li2CO3.
chromatogram obtained with the test solution, the area of any Description. A white, crystalline powder; odourless.
Identification
cent) and the sum of areas of all the secondary peaks is not A. When moistened with hydrochloric acid and introduced
more than twice the area of the peak in the chromatogram on a platinum wire, it imparts a red colour to a non-luminous
obtained with the reference solution (b) (2.0 per cent). flame.
Other tests. Comply with the tests stated under Tablets. B. Dissolve 0.2 g in 1 ml of hydrochloric acid and evaporate
to dryness on a water-bath; the residue is soluble in 3 ml of
ethanol (95 per cent).
Solvent mixture. A mixture of 4 volumes of water and 1 volume
C. Gives reaction A of carbonates (2.3.1).
of methanol.
Test solution. Transfer to a suitable size volumertric flask 10 Tests
tablets, add the solvent mixture to fill about half of the
Appearance of solution. Suspend 10.0 g in 30 ml of distilled
volumetric flask, shake the flask by mechanical means for 20
water and dissolve by adding 22 ml of nitric acid. Neutralise
minutes and dilute with solvent mixture which will yield a
with 2 M sodium hydroxide and dilute to 100.0 ml with distilled
solution having a concentration of about 0.02 per cent w/v
water (solution A). The solution is clear (2.4.1), and colourless
and filter.
(2.4.1).
Reference solution. A 0.02 per cent w/v solution of lisinopril
RS in solvent mixture.
Chromatographic system of bromine with a few drops of stannous chloride solution
– a stainless steel column 25 cm x 4.6 mm packed with AsT. The resulting solution complies with the limit test for
octylsilane bonded to porous silica (5 µm), (such as arsenic (2 ppm).
Symmetry C-8),
Calcium and magnesium. Dissolve 1.0 g in 30 ml of 1 M
hydrochloric acid and neutralise with dilute ammonia
– mobile phase: dissolve 1.0 g of hexane sulphonic
solution, filter, if necessary, and divide into two portions; to
acid sodium salt in 800 volumes of phosphate solution
one portion add 1 ml of ammonium oxalate solution; no
prepared by dissolving 4.1 g of monobasic potassium
turbidity or precipitate is produced on standing for 5 minutes.
phosphate in 900 ml water, adjust the pH to 2.0 with
To the other portion add 1 ml of disodium hydrogen phosphate
orthophosphoric acid, dilute to 1000 ml with water and
solution; no turbidity or precipitate is produced on standing
200 volumes of acetonitrile, mix, filter and degas.
for 5 minutes.
– spectrophotometer set at 215 nm, Heavy metals (2.3.13). Mix 1.0 g with 5 ml of water and 15 ml of
– a 20 µl loop injector. dilute hydrochloric acid, heat to boiling and maintain that
temperature for 1 minute. Add 1 drop of phenolphthalein
solution and sufficient ammonia solution to give the solution
column efficiency is not less than 850 theoretical plates, the
a faint pink colour. Cool and dilute to 25 ml with water. The
resulting solution complies with the limit test for heavy metals,
Method A ( 20 ppm).
Iron (2.3.14). 20 ml of solution A complies with the limit test for
Calculate the content of C21H31N3O5 iron (20 ppm).
690
IP 2007 LOMUSTINE
Potassium. Dissolve 1.0 g in 10 ml of 7 M hydrochloric acid, Other tests. Comply with the tests stated under Tablets.
add sufficient water to produce 50 ml and determine by flame Assay. Weigh and powder 20 tablets. Weigh accurately a
photometry (2.4.4), measuring at 766.5 nm, using potassium quantity of the powder containing about 1 g of Lithium
solution FP, suitably diluted with water, to prepare the Carbonate, add 100 ml of water and 50.0 ml of 1 M hydrochloric
standard solutions (500 ppm). acid and boil for 1 minute to remove carbon dioxide. Cool and
Sodium. Dissolve 1.0 g in 10 ml of 7 M hydrochloric acid, add titrate with 1 M sodium hydroxide using methyl orange
sufficient water to produce 50 ml and determine by flame solution as indicator.
photometry (2.4.4), measuring at 589 nm, using sodium
1 ml of 1 M hydrochloric acid is equivalent to 0.03695 g of
solution FP, suitably diluted with water, to prepare the
Li2CO3.
standard solutions (500 ppm).
Chlorides (2.3.12). 10 ml of solution A diluted to 15 ml with
water complies with the limit test for chlorides (250 ppm).
Sulphates (2.3.17). Disperse 0.75 g in 5 ml of distilled water
and dissolve by adding 5 ml of 7 M hydrochloric acid. Boil Lomustine
for 2 minutes, cool, neutralise with 2 M sodium hydroxide and
dilute to 25 ml with distilled water. The resulting solution
H NO
complies with the limit test for sulphates (200 ppm).
N N
Assay. Weigh accurately about 0.5 g, dissolve in 25.0 ml of Cl
1 M hydrochloric acid and titrate with 1 M sodium hydroxide O
using methyl orange solution as indicator.
1 ml of 1 M hydrochloric acid is equivalent to 0.03695 g of C9H16ClN3O2 Mol. Wt. 233.7
Li2CO3.
Lomustine is 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea.
Storage. Store protected from moisture. Lomustine contains not less than 98.5 per cent and not more
than 100.5 per cent of C9H16ClN3O2, calculated on the dried
basis.
Lithium Carbonate Tablets Description. A yellow, crystalline powder.
Lithium Carbonate Tablets contain not less than 95.0 per cent Carry out the tests protected from light and prepare the
and not more than 105.0 per cent of lithium carbonate, Li2CO3. solutions immediately before use.
A small quantity of the powdered tablets, when moistened Test A may be omitted if tests B, C and D are carried out. Tests
with hydrochloric acid and introduced on a platinum wire, B, C and D may be omitted if test A is carried out.
imparts a red colour to a non-luminous flame.
Tests Compare the spectrum with that obtained with lomustine RS
or with the reference spectrum of lomustine.
Apparatus. No 1 0.002 per cent w/v solution in ethanol (95 per cent) shows an
Medium. 900 ml of water absorption maximum at about 230 nm; absorbance at about
Speed and time. 100 rpm and 30 minutes. 230 nm, about 0.52.
Withdraw 90.0 ml of the medium, add a drop of hydrochloric C. In the test A for Related substances, the principal spot in
acid and dilute to 100.0 ml with water. Determine by flame the chromatogram obtained with test solution (b) corresponds
photometry Method A (2.4.4), or by atomic absorption to that in the chromatogram obtained with reference solution
spectrophotometry, Method A (2.4.2), measuring at 671 nm (c).
and using lithium solution FP, or lithium solution AAS, as D. Dissolve about 25 mg in 1 ml of methanol, add 0.1 ml of 2 M
appropriate, suitably diluted with water, for the standard sodium hydroxide and 2 ml of water and acidify by adding,
solution. dropwise, 1 M nitric acid. The resulting solution gives the
D. Not less than 60 per cent of the stated amount of Li2CO3. reactions of chlorides (2.3.1).
691
LOMUSTINE CAPSULES IP 2007
Tests – flow rate. 2 ml per minute,

Related substances. A. Determine by thin-layer – a 20 µl loop injector.
chromatography (2.4.17), coating the plate with silica gel G.
Inject the reference solution. The retention time of lomustine
Mobile phase. A mixture of 80 volumes of toluene and is about 25 minutes. When using a recorder, adjust the
20 volumes of glacial acetic acid. sensitivity of the system so that the height of the principal
Test solution (a). Dissolve 0.25 g of the substance under peak in the chromatogram obtained with the reference solution
examination in 10 ml of methanol. is not less than 50 per cent of the full scale of the recorder.
Test solution (b). Dissolve 0.1 g of the substance under Inject separately each solution. In the chromatogram obtained
examination in 100 ml of methanol. with the test solution the sum of areas of any secondary peaks
is not greater than the area of the peak in the chromatogram
Reference solution (a). A 0.01 per cent w/v solution of the obtained with the reference solution. Ignore any peak with an
substance under examination in methanol. area less than one-twentieth of that of the principal peak in
Reference solution (b). A 0.005 per cent w/v solution of the the chromatogram obtained with the reference solution.
substance under examination in methanol. Chlorides (2.3.12). Dissolve 0.24 g in 4 ml of methanol, add
Reference solution (c). A 0.1 per cent w/v solution of 20 ml of water, allow to stand for 20 minutes and filter. To 10 ml
lomustine RS in methanol. of the filtrate add 5 ml of methanol. The resulting solution
complies with the limit test for chlorides, replacing the 5 ml of
Reference solution (d). A solution containing 0.1 per cent water in the standard solution with 5 ml of methanol
w/v each of lomustine RS and 1,3-dicyclohexylurea in (0.25 per cent).
methanol.
Apply to the plate 5 µl of each solution. After development, on 1.0 g by drying over phosphorus pentoxide at a pressure
dry the plate, heat it at 110° for 1 hour, exposing the hot plate not exceeding 0.7 kPa for 24 hours.
in a closed tank containing chlorine, produced by adding dilute
hydrochloric acid to a 5 per cent w/v solution of potassium Assay. Weigh accurately about 0.2 g, add 20 ml of a 20 per cent
permanganate contained in a beaker placed at the bottom of w/v solution of potassium hydroxide and boil under a reflux
tank. Allow the plate to stand in contact with the chlorine condenser for 2 hours. Add 75 ml of water and 4 ml of nitric
vapours for 5 minutes. Remove the plate from the tank and dry acid, cool. Titrate with 0.05 M silver nitrate, determining the
it in a current of cold air until the excess of chlorine is removed end-point potentiometrically (2.4.25). Repeat the operation
without the substance under examination. The difference
and an area of the plate below the line of application produces
between the titrations represents the amount of silver nitrate
at most a very faint blue colour with 0.05 ml of potassium
required.
iodide and starch solution; avoid prolonged exposure to cold
air. Spray the plate with potassium iodide and starch solution. 1 ml of 0.05 M silver nitrate is equivalent to 0.01168 g of
Any secondary spot in the chromatogram obtained with test C9H16ClN3O2.
solution (d) shows two clearly separated principal spots. Lomustine Capsules
B. Determine by liquid chromatography (2.4.14). Lomustine Capsules contain not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of lomustine,
Test solution. Dissolve 0.25 g of the substance under C9H16ClN3O2.
Carry out the tests protected from light and prepare the
Reference solution. A 0.025 per cent w/v solution of the solutions immediately before use.
Identification
– a stainless steel column 20 cm x 4 mm, packed with Shake a quantity of the contents of the capsules containing
octadecylsilane bonded to porous silica (5 µm), 0.2 g of Lomustine with 10 ml of methanol, filter and evaporate
– mobile phase: equal volumes of methanol and water, the filtrate to dryness using a rotary evaporator on a water-
692
IP 2007 LOMUSTINE CAPSULES
bath maintained at not more than 60°. The residue, after drying Test solution. Shake a quantity of the contents of the capsules
at a pressure not exceeding 0.7 kPa at 60° for 30 minutes, containing 0.25 g of Lomustine with 10 ml of methanol and
complies with the following tests. filter.
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution. Dilute 1 volume of the test solution to 50
Compare the spectrum with that obtained with lomustine RS volumes with methanol.
or with the reference spectrum of lomustine. Chromatographic system
B. Melting range. 89° to 91° (2.4.21). – a stainless steel column 20 cm × 4 mm, packed with
Tests – mobile phase: equal volumes of methanol and water,
Related substances. A. Determine by thin-layer – spectrophotometer set at 230 nm,
chromatography (2.4.17), coating the plate with silica gel G. – a 20 µl loop injector.
Mobile phase. A mixture of 80 volumes of toluene and Inject the reference solution. The retention time of lomustine
20 volumes of glacial acetic acid. is about 25 minutes. When using a recorder, adjust the
Test solution (a). Shake a quantity of the contents of the sensitivity of the system so that the height of the principal
capsules containing 0.25 g of Lomustine with 10 ml of methanol peak in the chromatogram obtained with the reference solution
and filter. is not less than 50 per cent of the full scale of the recorder.
Test solution (b). Dilute 1 volume of test solution (a) to Inject separately each solution. In the chromatogram obtained
250 volumes with methanol. with the test solution the sum of areas of any secondary peaks
is not greater than the area of the peak in the chromatogram
Reference solution (a). Dilute 1 volume of test solution (b) to obtained with the reference solution. Ignore any peak with an
2 volumes with methanol. area less than one-twentieth of that of the principal peak in
Reference solution (b). A 0.005 per cent w/v solution of the the chromatogram obtained with the reference solution.
substance under examination in methanol. Uniformity of content. For capsules containing 10 mg or
less.
lomustine RS in methanol. Comply with the test stated under Capsules.
Reference solution (d). A solution containing 0.1 per cent Transfer the contents of a capsule quantitatively to a 100-ml
w/v each of lomustine RS and 1,3-dicyclohexylurea in volumetric flask with the aid of 20 ml of ethanol (95 per cent),
methanol. shake well, make up to the volume with ethanol (95 per cent)
and filter. Dilute suitably with the same solvent and measure
dry the plate, heat it at 110° for 1 hour, exposing the hot plate
about 230 nm (2.4.7). Calculate the content of C9H16ClN3O2 in
in a closed tank containing chlorine, produced by adding dilute
the capsule taking 260 as the specific absorbance at 230 nm.
hydrochloric acid to a 5 per cent w/v solution of potassium
permanganate contained in a beaker placed at the bottom of Other tests. Comply with the tests stated under Capsules.
tank. Allow the plate to stand in contact with the chlorine Assay. Weigh accurately a quantity of the mixed contents of
vapours for 5 minutes. Remove the plate and dry it in a current 20 capsules containing about 40 mg of Lomustine and shake
of cold air until the excess of chlorine is removed and an area with 70 ml of ethanol (95 per cent) for 20 minutes, dilute to
of the plate below the line of application produces at most a 100.0 ml with ethanol (95 per cent) and filter. Dilute 5.0 ml of
very faint blue colour with 0.05 ml of potassium iodide and the filtrate to 100.0 ml with ethanol (95 per cent) and measure
starch solution; avoid prolonged exposure to cold air. Spray the absorbance of the resulting solution at the maximum at
the plate with potassium iodide and starch solution. Any about 230 nm (2.4.7). Calculate the content of C9H16ClN3O2
secondary spot in the chromatogram obtained with test taking 260 as the specific absorbance at 230 nm.
chromatogram obtained with reference solution (a) and not Storage. Store protected from light and moisture.
solution (d) shows two clearly separated principal spots.
B. Determine by liquid chromatography (2.4.14).
693
LOPERAMIDE HYDROCHLORIDE IP 2007
Loperamide Hydrochloride Tests

O CH3 than reference solution BYS7 (2.4.1).
N
HO Related substances. Determine by liquid chromatography
CH3
, HCl (2.4.14).
Cl N
C29H33CIN2O2,HCl Mol. Wt. 513.5 w/v each of loperamide hydrochloride RS and haloperidol
Loperamide Hydrochloride is 4-(4-chlorophenyl)-4- RS in methanol.
hydroxypiperidino]-N,N-dimethyl-2,2-diphenylbutyramide Reference solution (b). Dilute 1 ml of the test solution to
hydrochloride. 100 ml with methanol. Dilute 5 ml of this solution to 20 ml with
Loperamide Hydrochloride contains not less than 99.0 per methanol.
cent and not more than 101.0 per cent of C29H33CIN2O2,HCl, Chromatographic system
calculated on the dried basis. – a stainless steel column 10 cm x 4.6 mm, packed with
Description. A white or almost white powder. octadecylsilane bonded to porous silica (5 µm),
– mobile phase: A. a 1.7 per cent w/v solution of
Identification tetrabutylammoniumhydrogen sulphate,
B. acetonitrile,
Test B may be omitted if tests A and C are carried out. Test A – flow rate. 2 ml per minute,
may be omitted if tests B and C are carried out. – a linear gradient programme using the conditions given
A. Determine by infrared absorption spectrophotometry (2.4.6). below,
Compare the spectrum with that obtained with loperamide – spectrophotometer set at 254 nm,
hydrochloride RS. – a 10 µl loop injector.
B. Determine by thin-layer chromatography (2.4.17), coating Time Mobile phase A Mobile phase B
the plate with octadecylsilyl silica gel. (min) (per cent v/v) (per cent v/v)
Mobile phase. A mixture of 40 volumes of dioxan, 40 volumes 0 90 10
of methanol and 20 volumes of ammonium acetate. 10 30 70
Test solution. Dissolve 0.6 g of the substance under 15 30 70
examination in 100 ml of the mobile phase. Equilibrate the column for at least 30 minutes with acetonitrile
Reference solution (a). A 0.6 per cent w/v solution of and then equilibrate at the initial eluent composition for at
loperamide hydrochloride RS in the mobile phase. least 5 minutes.
Reference solution (b). A solution containing 0.6 per cent Adjust the sensitivity of the detector so that the height of the
w/v each of loperamide hydrochloride RS and ketaconazole principal peak in the chromatogram obtained with reference
RS in the mobile phase. solution (b) is 70 per cent to 90 per cent of the full scale of the
recorder.
dry the plate in air for 15 minutes and expose it to iodine Inject reference solution (a) and record the peak responses.
vapours until the spots appear. The principal spot in the The retention times are: haloperidol, about 3 minutes and
chromatogram obtained with the test solution corresponds to loperamide hydrochloride, about 4.5 minutes. The test is not
that in the chromatogram obtained with reference solution (a). valid unless the resolution between the peaks corresponding
The test is not valid unless the chromatogram obtained with to haloperidol and loperamide hydrochloride is at least 8.0.
reference solution (b) shows two clearly separated spots. Inject separately methanol as a blank, the test solution and
C. Dissolve 50 mg in a mixture of 0.4 ml of strong ammonia reference solution (b). In the chromatogram obtained with the
solution and 2 ml of water. Mix, allow to stand for 5 minutes test solution, the area of any secondary peaks is not greater
and filter. Acidify the filtrate with 2 M nitric acid. It gives than the area of the principal peak in the chromatogram
reaction A of chlorides (2.3.1). obtained with reference solution (b) (0.5 per cent). Ignore any
694
IP 2007 LOPERAMIDE CAPSULES
peak with an area less than 0.2 times the area of the principal 1 M sodium hydroxide to a pH of 4.70 ± 0.05, diluting with
peak in the chromatogram obtained with reference solution water to 1000 ml and mixing
(b). Speed and time.100 rpm and 30 minutes.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Withdraw a suitable volume of the medium and filter.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Test solution. The filtrate from the dissolution medium.
Reference solution. A solution of loperamide hydrochloride
Assay. Weigh accurately about 0.4 g, dissolve in 25 ml of RS in the dissolution medium containing the same
anhydrous glacial acetic acid, add 10 ml of mercuric acetate concentration of loperamide hydrochloride as that expected
solution. Titrate with 0.1 M perchloric acid, using alpha- in the dissolution medium in the vessel.
naphtholbenzein solution as indicator. Carry out a blank
Determine by liquid chromatography (2.4.14) using the
titration.
chromatographic system described under Assay.
Calculate the content of C29H33CIN2O2,HCI.
C29H33CIN2O2,HCl.
C29H33ClN2O2, HCI.
Tablets.
Loperamide Capsules
Transfer the contents of one capsule to a 200-ml volumetric
Loperamide Hydrochloride Capsules flask. Add 35 ml of 0.5 M hydrochloric acid and mix with the
Loperamide Capsules contain not less than 90.0 per cent and aid of ultrasound for 15 minutes. Add 35 ml of acetonitrile
not more than 110.0 per cent of the labeled amount of and mix with the aid of ultrasound for another 15 minutes.
loperamide hydrochloride C29H33ClN2O2,HCI. Dilute to volume with a mixture of equal volumes of 0.5 M
hydrochloric acid and acetonitrile mix and filter.
Identification Determine by liquid chromatography (2.4.14) using the
A. Determine by thin-layer chromatography (2.4.17), coating chromatographic system and the reference solution described
the plate with silica gel G. under Assay.
Mobile phase. A mixture of 85 volumes of chloroform, Calculate the content of C29H33CIN2O2, HCI in the capsule.
10 volumes of methanol, and 5 volumes of formic acid. Assay. Determine by liquid chromatography (2.4.14).
Test solution. To a quantity of the contents of the capsules, Test solution. Transfer an accurately weighed portion of the
containing about 10 mg of Loperamide Hydrochloride, add mixed contents of 20 capsules containing about 20 mg of
10 ml of methanol, shake for 5 minutes, and filter. Loperamide Hydrochloride, to a 100-ml volumetric flask. Add
Reference solution. A 0.1 per cent w/v solution of loperamide about 35 m of 0.5 M hydrochloric acid and mix with the aid of
hydrochloride RS in methanol. ultrasound for 15 minutes. Add 35 ml of acetonitrile and mix
with the aid of ultrasound for an additional 15 minutes. Dilute
Apply to the plate 10 µl of each solution. After development, with a mixture of equal volumes of acetonitrile and 0.5 M
dry the plate in air and expose to iodine vapour. The principal hydrochloric acid, mix, and filter. Transfer 5.0 ml of this solution
spot in the chromatogram obtained with the test solution to a 100-ml volumetric flask, dilute to volume with the same
corresponds to that in the chromatogram obtained with the solvent mixture and mix.
reference solution.
B. In the Assay, the principal peak in the chromatogram loperamide hydrochloride RS in a mixture of equal volumes
obtained with the test solution corresponds to the peak in the of acetonitrile and water.
Tests – a stainless steel column 25 cm x 4 mm, packed with nitrile
groups chemically bonded to porous silica particles
Dissolution (2.5.2)
(10 µm),
Apparatus No. 2 – mobile phase: dilute 500 ml of acetonitrile to 1000.0 ml
Medium. 500 ml of pH 4.7 acetate buffer, prepared by mixing with water, add 20 drops of phosphoric acid, mix, and
200 ml of 1 M acetic acid with 600 ml of water, adjusting with filter,
695
LOPERAMIDE TABLETS IP 2007
– flow rate. 2 ml per minute, D. Not less than 80 per cent of the stated amount of
– spectrophotometer set at 220 nm, C29H33ClN2O2, HCI.
– a 50 µl loop injector. Uniformity of content. Comply with the test stated under
Inject the reference solution. The column efficiency, determined Tablets.
from the analyte peak is not less than 1900 theoretical plates, Test solution. Crush 1 tablet and transfer to a 200-ml volumetric
the capacity factor, is not less than 3.5, and the relative standard flask. Add 4 ml of a 5 per cent w/v solution of phosphoric acid
deviation for replicate injections is not more than 2.0 per cent. and 20 ml of methanol shake and dilute to volume with water.
Inject alternately the test solution and the reference solution. Reference solution. Dissolve an accurately weighed quantity
Calculate the content of C29H33CIN2O2,HCI in the capsules. of loperamide hydrochloride RS in methanol to obtain a
Quantitatively dilute this solution with water to obtain a
solution having a known concentration of about 0.2 mg per
ml. To 10.0 ml of this solution add 5.0 ml of 5 per cent
Loperamide Tablets phosphoric acid solution and 25 ml of methanol, dilute with
water to 200.0 ml and mix.
Loperamide Hydrochloride Tablets
Determine by liquid chromatography (2.4.14) using the
Loperamide Hydrochloride Tablets contain not less than
chromatographic system described under Assay.
90.0 per cent and not more than 110.0 per cent of the labeled
amount of loperamide hydrochloride C29H33CIN2O2,HCI. Calculate the content of C29H33CIN2O2, HCI in the tablet.
Identification
Buffer solution. To 3.0 g of triethylamine hydrochloride and
A. Transfer a quantity of finely powdered tablets containing 1.0 ml of phosphoric acid add 550 ml of water and mix.
about 10 mg of Loperamide Hydrochloride, to a test-tube, add Test solution. Weigh and finely powder 20 Tablets. Transfer
20.0 ml of isopropyl alcohol. shake by mechanical means for an accurately weighed quantity of the powder containing about
one minute, and allow to settle. Dilute 9.0 ml of the supernatant 16 mg of Loperamide Hydrochloride, to a 200-ml volumetric
to 10 ml with 0.1 M hydrochloric acid. The solution so flask. Add 4 ml of a 5 per cent w/v solution of phosphoric acid
obtained shows absorption maxima and minima at the same and 20 ml of methanol, dilute with water to volume, and mix.
wavelengths as that of a similar preparation of loperamide
hydrochloride RS. Reference solution. Dissolve an accurately weighed quantity
of loperamide hydrochloride RS in methanol to obtain a
B. In the Assay, the principal peak in the chromatogram solution having a known concentration of about 2 mg per ml.
obtained with the test solution corresponds to the peak in the Quantitatively dilute this solution with water to obtain a
chromatogram obtained with the reference solution. solution having a known concentration of about 0.2 mg per
ml. Transfer 10.0 ml of this solution to a 250-ml volumetric
Tests flask, add 5.0 ml of 5 per cent phosphoric acid solution and
25 ml of methanol, dilute with water to volume, and mix.
Apparatus No. 1
Medium. 900 of 0.01 M hydrochloric acid octylsilane chemically bonded to totally porous silica
Speed and time. 50 rpm and 30 minutes. particles (5 µm),
Withdraw 10 ml of the medium from each vessel, mix and filter. – mobile phase: a mixture of 45 volumes of acetonitrile
and 55 volumes of buffer solution,
Test solution. The mixed filtrate from the dissolution medium. – flow rate. 2 ml per minute,
Reference solution. A solution of loperamide hydrochloride – spectrophotometer set at 214 nm,
RS in the dissolution medium containing the same – a 20 µl loop injector.
concentration of loperamide hydrochloride as that expected Inject the reference solution. The tailing factor is not more
in the dissolution medium in the vessel. than 2.0; and the relative standard deviation for replicate
Determine by liquid chromatography (2.4.14) using the injections is not more than 2.0 per cent.
chromatographic system described under Assay. Inject alternately the test solution and the reference solution.
Calculate the content of C29H33CIN2O2, HCI in the medium. Calculate the content of C29H33CIN2O2,HCI in the tablets.
696
IP 2007 LOPINAVIR
Lopinavir – mobile phase: gradient mixtures of acetonitrile and the

buffer (pH 2.5),
H3C CH3 below,
H O CH3 – spectrophotometer set at 210 nm,
N O – a 20 µl loop injector.
N N
O OH H Time Buffer (pH 2.5) Acetonitril
N O H3C (in min.) (per cent v/v) (per cent v/v)
H 0 70 30
30 30 70
C37H48N4O5 Mol. Wt. 628.8 40 20 80
Lopinavir is (αS)-N-[(1S,3S,4S)-4-[[(2,6- 50 20 80
dimethylphenoxy)acetyl]amino-3-hydroxy-5-phenyl-1-
(phenylmethyl)pentyl]tetrahydro-α-(1-methylethyl)-2-oxo-1- Inject the reference solution. The test is not valid unless the
(2H)-pyrimidineacetamide. column efficiency determined from the lopinavir peak is not
less than 5000 theoretical plates and the tailing factor is not
Lopinavir contains not less than 98.0 per cent and not more more than 1.5.
than 102.0 per cent of C37H48N4O5, calculated on the anhydrous
basis. Inject separately the test solution and the reference solution.
Any secondary peak should not be more than 0.3 per cent and
Description. A white or almost white powder. the sum of the areas of all the secondary peaks should not be
more than 1.0 per cent when calculated by percentage area
Identification
normalisation.
A. Determine by infrared absorption spectrophotometry (2.4.6). Heavy metals (2.3.13). 1.0 g complies with limit test for heavy
Compare the spectrum with that obtained with lopinavir RS metals, Method B (20 ppm).
or with the reference spectrum of lopinavir.
obtained with the test solution corresponds to the peak due Water (2.3.43). Not more than 4.0 per cent, determined on
to lopinavir in the chromatogram obtained with the reference 0.1 g.
solution. Assay. Determine by liquid chromatography (2.4.14).
Tests Test solution. Dissolve about 50 mg, accurately weighed, of

the substance under examination in a suitable quantity of
Specific optical rotation (2.4.22). –22.0° to –26.0°, determined solution A in a 50-ml volumetric flask and dilute to volume
in a 0.4 per cent w/v solution in methanol and calculated on with solution A. Dilute 10.0 ml of this solution to 50.0 with
the anhydrous basis. solution A.
Related substances. Determine by liquid chromatography Reference solution. Dissolve about 50 mg, accurately weighed,
(2.4.14). of lopinavir RS in a suitable quantity of solution A in a 50-ml
Solvent mixture. 70 volumes of a buffer and 30 volumes of volumetric flask and dilute to volume with solution A. Dilute
acetonitrile, the buffer being prepared by dissolving 2.72 g of 10.0 ml of this solution to 50.0 ml with solution A.
potassium dihydrogen phosphate in 900 ml of water, pH of Chromatographic system
which is adjusted to 2.5 with phosphoric acid, and diluting to – a stainless steel column 25 cm x 4.6 mm, packed with
1000 ml with water (solution A). octylsilane bonded to porous silica (5 µm),
Test solution. Dissolve 0.1 g of the substance under – mobile phase: 55 volumes of solution B prepared by
examination in 100 ml of solvent mixture. mixing 80 volumes of acetonitrile and 20 volumes of
methanol and 45 volumes of 0.05 M potassium
Reference solution. A 0.01 per cent w/v solution of lopinavir dihydrogen phosphate, the pH of which is adjusted to
RS in solvent mixture. 3.0 with dilute phosphoric acid,
Chromatographic system – flow rate. 1.5 ml per minute,
– a stainless steel column 25 cm x 4.6 mm, packed with – spectrophotometer set at 210 nm,
octylsilane bonded to porous silica (5 µm), – a 20 µl loop injector.
697
LOPINAVIR AND RITONAVIR CAPSULES IP 2007
Inject the reference solution. The test is not valid unless the Test solution. Weigh accurately a quantity of the contents of
column efficiency determined from the lopinavir peak is not the capsules containing 100 mg of Lopinavir, disperse in 100
less than 1500 theoretical plates, the tailing factor is not more ml of the solvent mixture and filter.
than 2.0 and the relative standard deviation for replicate Reference solution (a). A 0.1 per cent w/v solution of lopinavir
injections is not more than 2.0 per cent. RS in the solvent mixture.
Inject separately the test solution and the reference solution
and measure the responses for the principal peak.
100 ml with the solvent mixture.
Storage. Store protected from light and moisture. – a stainless steel column 25 cm x 4.6 mm, packed with
octadecylsilane bonded to porous silica (5µm),
– mobile phase: A. a mixture of 45 volumes of acetonitrile
Lopinavir and Ritonavir Capsules and 55 volumes of a buffer solution prepared by
dissolving 1.36 g of potassium dihydrogen
Lopinavir and Ritonavir Capsules contain not less than 90.0 orthophosphate dihydrate in 1000 ml of water and
per cent and not more than 110.0 per cent of the stated amounts adjusting the pH to 4.0 with orthophosphoric acid,
of lopinavir, C37H48N4O5 and ritonavir, C37H48N6O5S2. B. a mixture of 80 volumes of acetonitrile
and 20 volumes of the buffer solution,
Identification
In the Assay, the principal peaks in the chromatogram obtained – a linear gradient programme using the conditions given
with the test solution correspond to the peaks in the below,
chromatogram obtained with the reference solution. – spectrophotometer set at 210 nm,
Tests
Dissolution (2.5.2). (in min) (per cent v/v) (per cent v/v)
Apparatus. No 1 0 100 0
Medium. 900 ml of a solution prepared by dissolving 15.7 g of 80 100 0
polyoxyethylene 10-lauryl ether in 1000 ml of a 0.85 per cent 90 0 100
v/v solution of hydrochloric acid. 120 0 100
Speed and time. 75 rpm and 120 minutes 130 100 0
Withdraw a suitable volume of the medium and filter. 140 100 0
Determine by liquid chromatography (2.4.14) Inject reference solution (a). The test is not valid unless the
Test solution. The filtrate obtained as given above. Dilute the column efficiency is not less than 2000 theoretical plates and
filtrate if necessary, with the dissolution medium. the tailing factor is not more than 2.0.
Reference solution. A solution containing 0.15 per cent w/v of Inject the test solution and reference solution (b). In the
lopinavir RS and 0.04 per cent w/v of ritonavir RS in chromatogram obtained with the test solution, the area of any
methanol. Dilute 5 ml of the solution to 50 ml with the secondary peak is not more than the area of the peak in the
dissolution medium. chromatogram obtained with the reference solution (b)
Use the chromatographic system given in the Assay.
Inject the test solution and the reference solution. obtained with the reference solution (b) (2.0 per cent).
Calculate the contents of C37H48N4O5 and C37H48N6O5S2. For Ritonavir
D. Not less than 75 per cent of the stated amounts of Solvent mixture. 40 volumes of the buffer solution and
C37H48N4O5 and C37H48N6O5S2. 60 volumes of acetonitrile.
Related substances. Determine by liquid chromatography Test solution. Weigh accurately a quantity of the contents of
(2.4.14). the capsules containing 50 mg of Ritonavir, disperse in 100 ml
For Lopinavir of the solvent mixture and filter.
Solvent mixture. 40 volumes of buffer solution and 60 volumes Reference solution (a). A 0.05 per cent w/v solution of
of acetonitrile. ritonavir RS in the solvent mixture.
698
IP 2007 LOPINAVIR AND RITONAVIR TABLETS
Reference solution (b). Dilute 1 ml of reference solution (a) to Chromatographic system

100 ml with the solvent mixture. – a stainless steel column 5 cm x 4.6 mm, packed with
Chromatographic system octadecylsilane bonded to porous silica ( 5 µm),
– a stainless steel column 15 cm x 4.6 mm, packed with – mobile phase: a mixture of 45 volumes of buffer solution
silica gel consisting of porous spherical particles with prepared by dissolving 6.8 g of potassium dihydrogen
chemically bonded butyl group ( 3µm) (such as YMC orthophosphate dihydrate in 1000 ml with water and
C4), adjusting the pH to 3.0 with orthophosphoric acid, 42.5
– column temperature 60º, volumes of acetonitrile and 12.5 volumes of methanol,
– mobile phase: A. a mixture of 69 volumes of buffer – flow rate. 1 ml per minute,
solution prepared by dissolving 4.1 g of monobasic – spectrophotometer set at 210 nm,
potassium phosphate in 1000 ml of distilled water and – a 10 µl loop injector.
filtering, and 18 volumes of acetonitrile, 8 volumes of Inject the reference solution. The test is not valid unless the
tetrahydrofuran and 5 volumes of n-Butanol. resolution between lopinavir and ritonavir peak is not less
B. a mixture of 40 volumes of the buffer than 2.5, the column efficiency for each component is not less
solution, 47 volumes of acetonitrile, 8 volumes of than 2000 theoretical plates, the tailing factor is not more than
tetrahydrofuran and 5 volumes of n-Butanol, 2.0 and the relative standard deviation for replicate injections
– flow rate. 1 ml per minute, is not more than 2.0 per cent.
– a linear gradient programme using the conditions given Inject the test solution and the reference solution.
below,
Calculate the contents of C37H48N4O5 and C37H48N6O5S2 in the
capsules.
Time Mobile phase A Mobile phase B Storage. Store protected from moisture in a refrigerator (2º to
(in min) (per cent v/v) (per cent v/v) 8º).
0 100 0
60 100 0
120 0 100 Lopinavir and Ritonavir Tablets
130 100 0 Lopinavir and Ritonavir Tablets contains not less than 90.0
155 100 0 per cent and not more than 110.0 per cent of the stated amounts
of lopinavir, C37H48N4O5 and ritonavir, C37H48N6O5S2.
Identification
Inject the test solution and reference solution (b). In the In the Assay, the principal peaks in the chromatogram obtained
chromatogram obtained with the test solution, the area of any with test solution correspond to the principal peaks in the
secondary peak is not more than 2.5 times the area of the peak chromatogram obtained with the reference solution.
(2.5 per cent) and the sum of areas of all the secondary peaks Tests
is not more than 5 times the area of the peak in the Dissolution (2.5.2).
chromatogram obtained with the reference solution (b) (5.0
per cent). Apparatus. No 1
Medium. 900 ml of a solution prepared by dissolving 15.7 g of
Other tests. Comply with the tests stated under Capsules. polyoxyethylene 10 lauryl ether in 1000 ml of a 0.85 per cent
Assay. Determine by liquid chromatography (2.4.14). v/v solution of hydrochloric acid.
Test solution. Mix the contents of 20 capsules. Weigh Speed and time. 75 rpm and 120 minutes
accurately a quantity of the mixed contents containing 70 mg Withdraw a suitable volume of the medium and filter.
of Lopinavir and disperse in 100.0 ml of methanol and filter.
Determine by liquid chromatography (2.4.14)
Dilute 5.0 ml of the filtrate to 50.0 ml with the mobile phase.
Test solution. The filtrate obtained as given above. Dilute the
filtrate if necessary, with the dissolution medium.
of lopinavir RS and 0.0175 per cent w/v of ritonavir RS in
methanol. Dilute 10.0 ml of the solution to 100.0 ml with the Reference solution. A solution containing 0.22 per cent w/v of
mobile phase. lopinavir RS and 0.055 per cent w/v of ritonavir RS in
699
LOPINAVIR AND RITONAVIR TABLETS IP 2007
methanol. Dilute 5 ml of the solution to 50 ml with the Reference solution (a). A 0.1 per cent w/v solution of ritonavir
dissolution medium. RS in the solvent mixture.
Use the chromatographic system described under Assay. Reference solution (b). Dilute 1 ml of reference solution (a) to
100 ml with the solvent mixture.
relative standard deviation for replicate injections is not more Chromatographic system
than 2.0 per cent. – a stainless steel column 15 cm x 4.6 mm, packed with
silica gel consisting of porous spherical particles with
chemically bonded with butyl group (3µm) (such as YMC
D. Not less than 70 per cent of the stated amounts of C4),
C37H48N4O5 and C37H48N6O5S2. – column temperature 60º,
Related substances. Determine by liquid chromatography – mobile phase: A. a mixture of 69 volumes of a buffer
(2.4.14). solution prepared by dissolving 4.1 g of potassium
dihydrogen phosphate in 1000 ml of water, 18 volumes
For Lopinavir of acetonitrile, 8 volumes of tetrahydrofuran and
Test solution. Disperse accurately a quantity of the powdered 5 volumes of n- butanol,
tablets containing 100 mg of Lopinavir in 100 ml of the mobile B. a mixture of 40 volumes of buffer
phase. solution, 47 volumes of acetonitrile, 8 volumes of
tetrahydrofuran and
5 volumes of n-butanol,
lopinavir RS in the mobile phase.
– flow rate.1 ml per minute,
Reference solution (b). Dilute 1 ml of reference solution (a) to – a linear gradient programme using the conditions given
100 ml with the mobile phase. below,
Chromatographic system – spectrophotometer set at 240 nm,
– a stainless steel column 25 cm x 4.6 mm, packed with – a 50 µl loop injector.
octadecylsilane bonded to porous silica (5mm), Time Mobile phase A Mobile phase B
– mobile phase: a mixture of 55 volumes of a buffer solution (in min.) (per cent w/v) (per cent w/v)
prepared by dissolving 1.36 g of potassium dihydrogen 0 100 0
orthophosphate in 1000 ml of water and adjusting the
60 100 0
pH to 4.0 with orthophosphoric acid, and 45 volumes
of acetonitrile, 120 0 100
– flow rate.1.5 ml per minute, 121 100 0
– spectrophotometer set at 210 nm, 135 100 0
Inject reference solution (a). The test is not valid unless the column efficiency is not less than 3000 theoretical plates and
column efficiency is not less than 3000 theoretical plates and the tailing factor is not more than 2.0.
Inject the test solution and reference solution (b). In the chromatogram obtained with the test solution, the area of any
chromatogram obtained with the test solution, the area of any secondary peak is not more than 2.5 times the area of the peak
secondary peak is not more than the area of the peak in the in the chromatogram obtained with the reference solution (b)
chromatogram obtained with the reference solution (b) (1.0 (2.5 per cent) and the sum of areas of all the secondary peaks
per cent) and the sum of areas of all the secondary peaks is is not more than 5 times the area of the peak in the
not more than twice the area of the peak in the chromatogram chromatogram obtained with the reference solution (b)
obtained with the reference solution (b) (2.0 per cent). (5.0 per cent).
For Ritonavir Other tests. Comply with the tests stated under Tablets.
Solvent mixture. A mixture of 40 volumes of a buffer solution
prepared by dissolving 4.1 g of potassium dihydrogen
0.5 g.
phosphate in 1000 ml of water, and 60 volumes of acetonitrile,
Test solution. Disperse an accurately weighed quantity of the
powdered tablets containing 100 mg of Ritonavir in 100 ml of Test solution. Weigh and powder 20 tablets. Dissolve an
the solvent mixture. accurately weighed quantity of the powder containing 200 mg
700
IP 2007 LOSARTAN POTASSIUM
of Lopinovir in 250.0 ml of methanol. Dilute 5.0 ml of the Losartan Potassium contains not less than 98.0 per cent and
resulting solution to 50.0 ml with the mobile phase. not more than 102.0 per cent of C22H22ClKN6O, calculated on
the anhydrous basis.
Reference solution. 5.0 ml of each of a 0.08 per cent w/v
solution of lopinavir RS and a 0.02 per cent w/v solution of Description. A white to off-white crystalline powder.
ritonavir RS in methanol, diluted to 50.0 ml with the mobile
phase. Identification
– a stainless steel column 15 cm x 4.6 mm, packed with Compare the spectrum with that obtained with losartan
octylsilane bonded to porous silica (5 µm), potassium RS or with the reference spectrum of losartan
– mobile phase: A. a mixture of 80 volumes of acetonitrile potassium.
and 20 volumes of methanol, B. When examined in the range 200 to 400 nm (2.4.7), a 0.001
B. a mixture of 45 volumes of a buffer per cent w/v solution of methanol and compares with the
solution prepared by dissolving 6.8 g of potassium absorbance obtained with a solution of losartan potassium
dihydrogen orthophosphate anhydrous in 1000 ml of RS prepared in a similar manner.
water and adjusting the pH to 3.0 with orthophosphoric
acid, and 55 volumes of mobile phase A. C. Gives reaction A of potassium (2.3.1).
Tests
– a 20 µl loop injector. Related substances. Determine by liquid chromatography
Inject the reference solution. The test is not valid unless the (2.4.14).
resolution between the peak due to lopinavir (retention time, Test solution. Dissolve 30 mg of the substance under
about 6 minutes) and the peak due to ritonavir (retention time, examination in 100 ml of methanol.
about 5 minutes) is less than 2.0 and the relative standard
Reference solution (a). A 0.03 per cent w/v solution of losartan
potassium RS and 0.0002 per cent of triphenylmethanol in
Inject the test solution and the reference solution. methanol.
Calculate the contents of C37H48N4O5 and C37H48N6O5S2. in the Reference solution (b). A 0.0003 per cent w/v solution of
tablets. losartan potassium RS in methanol.
Storage. Store protected from moisture, at a temperature not Chromatographic system
exceeding 30º. – a stainless steel column 25 cm x 4.0 mm packed with
– mobile phase: A. 0.1 per cent solution of
orthophosphoric acid in water and filter,
Losartan Potassium B. acetonitrile,
below,
Cl – flow rate. 1 ml per minute,
H3 C N – spectrophotometer set at 220 nm,
OH – a 10 µl loop injector.
N N N
Time Orthophosphoricacid Acetonitrile
K N N (in min.) ( per cent v/v) ( per cent v/v)
0 75 25
35 10 90
50 75 25
Inject reference solution (a). The relative retention times are
C22H22ClKN6O Mol. Wt. 461.0 about 1.0 for losartan and 1.9 for triphenylmethanol and the
tailing factor for losartan is not more than 2.0.
Losartan Potassium is monopotassium salt of 4-butyl-4-
chloro-1-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]-4-yl]methyl]- Inject the test solution and reference solution (b). In the
1H-imidazole-5-methanol. chromatogram obtained with the test solution, the area of any
701
LOSARTAN TABLETS IP 2007
secondary peak is not more than 0.5 times the area of the peak Identification
per cent) and the sum of areas of all the secondary peaks is In the Assay, the principal peak in the chromatogram obtained
not more than the area of the peak in the chromatogram with the test solution corresponds to the peak in the
obtained with the reference solution (b) (1.0 per cent). chromatogram obtained with the reference solution.
Heavy metals (2.3.13). 1.0 g complies with limit test for heavy Tests
1.0 g. Apparatus. No 1
Assay. Determine by liquid chromatography (2.4.14). Medium. 900 ml of water.
Test solution. Dissolve 25 mg of the substance under Speed and time. 50 rpm for 45 minutes.
examination in 100.0 ml of methanol. Withdraw a suitable volume of the medium and filter. Measure
Reference solution. A 0.025 per cent w/v solution of losartan the absorbance of the filtered solution, suitably diluted with
potassium RS in methanol. the medium if necessary, at the maximum at about 250 nm
(2.4.7). Calculate the content of C22H22ClKN6O in the medium
Chromatographic system from the absorbance obtained from a solution of known
– a stainless steel column 25 cm x 4.0 mm packed with concentration of losartan potassium RS in the same medium.
– mobile phase: A. 0.1 per cent w/v solution of D. Not less than 75 per cent of the stated amount of
orthophosphoric acid in water and filter, C22H22ClKN6O.
B. acetonitrile, Related substances. Determine by liquid chromatography
– a linear gradient programme using the conditions given (2.4.14).
below,
a quantity of the powdered tablet containing 100 mg of Losartan
Potassium, disperse in 100 ml of water and filter.
Time Orthophosphoric acid Acetonitrile Reference solution (a). A 0.1 per cent w/v solution of losartan
(in min.) ( per cent v/v) ( per cent v/v) potassium RS in water.
0 75 25 Reference solution (b). Dilute 1 ml of reference solution (a) to
100 ml with water.
35 10 90
50 75 25 Chromatographic system
Inject the reference solution. The test is not valid unless the octasilyl chemically bonded to porous silica (5 µm), (such
tailing factor is not more than 1.5, the column efficiency in not as Lichrosphere RP8e),
less than 5000 theoretical plates. The relative standard – mobile phase: a mixture of 75 volumes of buffer solution
deviation for replicate injections is not more than 2.0 per cent. prepared by mixing 770 mg of ammonium acetate in
Inject the test solution and the reference solution. 1000 ml of water, add 2.0 ml of triethylamine, adjust pH
to 6.5 with glacial acetic acid and 25 volumes of
Calculate the content of C22H22ClKN6O.
acetonitrile,
Storage. Store protected from moisture. – flow rate. 1.5 ml per minute,
Losartan Tablets tailing factor is not more than 3.0 and the column efficiency in
not less than 1000 theoretical plates.
Losartan Potassium Tablets
Losartan Tablets contain Losartan Potassium.
Losartan Tablets contain not less than 90.0 per cent and not secondary peak is not more than the area of the peak in the
more than 110.0 per cent of the stated amount of losartan chromatogram obtained with reference solution (b) (1.0 per
potassium, C22H22ClKN6O. cent) and the sum of areas of all the secondary peaks is not
702
IP 2007 LYNOESTRENOL
more than 2 times the area of the peak in the chromatogram Lynoestrenol contains not less than 98.0 per cent and not
obtained with the reference solution (b) (2.0 per cent). more than 102.0 per cent of C20H28O, calculated on the dried
Other tests. Comply with the tests stated under Tablets. basis.
Test solution. Transfer intact tablets in a suitable volumetric Identification
flask, dissolve in mobile phase and disperse completely. Dilute
with mobile phase to obtain a final concentration of 0.0125 per
and C may omitted if test A is carried out.
cent w/v.
Reference solution. A 0.125 per cent w/v solution of losartan
Compare the spectrum with that obtained with lynoestrenol
potassium RS in mobile phase. Dilute 10.0 ml of the solution to
RS .
100.0 ml with mobile phase.
B. In the test of Related substances, the principal spot in the
octasilyl chemically bonded to porous silica (5 µm), (such
as Lichrosphere RP8e) C. Melting range. 161° to 165° (2.4.21).
ammonium acetate, 30 volumes of acetonitrile,
Tests
5 volumes of methanol and 0.2 volumes of Appearance of solution. A 2.0 per cent w/v solution in ethanol
triethylamine, adjust the pH to 6.6 with glacial acetic (95 per cent) is clear (2.4.1), and colourless (2.4.1).
acid and filter,
in a 3.6 w/v solution in ethanol (95 per cent).
– a 20 µl loop injector. Related substances. Determine by thin-layer chromatography
tailing factor is not more than 2.0, the column efficiency is not Mobile phase. A mixture of 80 volumes of n-heptane and
less than 5000 theoretical plates and the relative standard 20 volumes of acetone.
deviation for replicate injections is not more than 2 per cent. Test solution (a). Dissolve 0.5 g of the substance under
Inject the test solution and the reference solution. examination in 100 ml of chloroform.
Calculate the content of C22H22ClKN6O. Test solution (b). Dissolve 0.25 g of the substance under
Labelling. The label states the strength of Losartan substance under examination in chloroform.
Potassium.
lynoestrenol RS in chloroform.
Lynoestrenol dry the plate in air and spray with 0.25 M ethanolic sulphuric
acid, heat at 105° for 10 minutes and examine in ultraviolet
Lynestrenol light at 365 nm. Any secondary spot in the chromatogram
H3C OH spot in the chromatogram obtained with reference solution
C CH (a).
H H Loss on drying (2.4.19). Not more than 0.5 per cent, determined
on 0.5g by drying in an oven at 105°.
H H
tetrahydrofuran, add 5 ml of a 10 per cent w/v solution of
silver nitrate. Titrate with 0.1 M sodium hydroxide,
C20H28O Mol. Wt. 284.4
Lynoestrenol is 19-nor-17α-pregn-4-en-20-yn-17β-ol. out a blank titration.
703
LYNOESTRENOL IP 2007

C20H28O.
704
M
Magaldrate ....
Magaldrate Oral Suspension ....
Magaldrate Tablets ....
Heavy Magnesium Carbonate ....
Light Magnesium Carbonate ....
Magnesium Chloride ....
Magnesium Hydroxide ....
Magnesium Hydroxide Oral Suspension ....
Heavy Magnesium Oxide ....
Light Magnesium Oxide ....
Magnesium Stearate ....
Magnesium Sulphate ....
Magnesium Trisilicate ....
Mannitol ....
Mannitol Injection ....
Mebendazole ....
Mebendazole Tablets ....
Mebeverine Hydrochloride ....
Mebeverine Tablets ....
Meclizine Hydrochloride ....
Meclizine Tablets ....
Medroxyprogesterone Acetate ....
Mefenamic Acid ....
Mefenamic Acid Capsules ....
Megestrol Acetate ....
Megestrol Tablets ....
Melphalan ....
Melphalan Injection ....
Melphalan Tablets ....
Menadione ....
705
Menthol
Mephentermine Sulphate
Mephentermine Injection
Mepyramine Maleate
Mepyramine Tablets
Meropenem
Meropenem Injection
Mercaptopurine
Mercaptopurine Tablets
Mestranol
Metformin Hydrochloride
Metformin Tablets
Methadone Hydrochloride
Methadone Injection
Methadone Tablets
Methdilazine Hydrochloride
Methdilazine Tablets
Methotrexate
Methotrexate Injection
Methotrexate Tablets
Methoxamine Hydrochloride
Methoxamine Injection
Methyl Salicylate
Industrial Methylated Spirit
Methylcellulose
Methyldopa
Methyldopa Tablets
Methylergometrine Maleate
Methylergometrine Injection
Methylergometrine Tablets
Methylparaben
Methylprednisolone
Methylprednisolone Tablets
706
Methylprednisolone Acetate ....

Methylprednisolone Acetate Injection ....
Metoclopramide Hydrochloride ....
Metoclopramide Injection ....
Metoclopramide Syrup ....
Metoclopramide Tablets ....
Metoprolol Tartrate ....
Metoprolol Tablets ....
Metronidazole ....
Metronidazole Benzoate ....
Metronidazole Benzoate Oral Suspension ....
Metronidazole Injection ....
Metronidazole Tablets ....
Mexiletine Hydrochloride ....
Mexiletine Capsules ....
Mexiletine Injection ....
Mianserin Hydrochloride ....
Mianserin Tablets ....
Miconazole Nitrate ....
Miconazole Cream ....
Miconazole Pessaries ....
Microcrystalline Cellulose ....
Microcrystalline Wax ....
Minoxidil ....
Minoxidil Tablets ....
Monothioglycerol ....
Morphine Sulphate ....
Morphine And Atropine Injection ....
Morphine Injection ....
Multiple Electrolytes And Dextrose Injection Type I ....
Multiple Electrolytes And Dextrose Injection Type II ....
Multiple Electrolytes And Dextrose Injection Type III ....
Multiple Electrolytes And Dextrose Injection Type IV ....
707
Multiple Electrolytes And Dextrose Injection Type V ....

Multiple Electrolytes Injection Type VI ....
Mustine Hydrochloride ....
Mustine Injection ....
708
IP 2007 MAGALDRATE
obtain a clear solution. Dilute 10.0 ml of the filtrate with water

Magaldrate to 100.0 ml. The emission intensity of this solution, determined
Al5Mg10(OH)31(SO4)2,xH2O Mol. Wt. 1097.4 (anhydrous) by flame photometry (2.4.4), at about 589 nm and corrected for
background transmission at about 580 nm, is not greater than
Magaldrate is a chemical combination of aluminium and
that produced by treating similarly a standard solution
magnesium hydroxides and sulphates corresponding
containing 2.2 µg of Na per ml.
approximately to the formula Al5Mg10(OH)31(SO4)2,xH2O.
Aluminium hydroxide. 32.1 to 45.9 per cent of Al(OH)3,
Magaldrate contains not less than 90.0 per cent and not more
calculated on the dried basis and determined by the following
than 105.0 per cent of Al5Mg10(OH)31(SO4)2, calculated on the
method. Dissolve about 100 mg, accurately weighed, in 3 ml of
dried basis.
dilute hydrochloric acid and dilute to 30 ml with water. Add
Description. A white or almost white, crystalline powder; with stirring 25.0 ml of 0.05 M disodium edetate, mix and allow
odourless. to stand for 5 minutes. Add 20 ml of acetic acid-ammonium
acetate buffer, 60 ml ethanol (95 per cent) and 2 ml of dithizone
Identification solution and titrate with 0.05 M zinc sulphate to a bright
A. Dissolve 0.8 g in 20 ml of 3 M hydrochloric acid, dilute rose-pink colour. Repeat the procedure without the substance
with water to 50 ml, add 3 drops of methyl red solution and under examination. The difference between the titrations
heat to boiling. Add dilute ammonia solution until the colour represents the amount of disodium edetate required.
changes to just yellow, continue boiling for 2 minutes and 1 ml of 0.05 M disodium edetate is equivalent to 0.0039 g of
filter; the filtrate gives the reactions of magnesium salts (2.3.1). Al(OH)3.
B. Wash the precipitate obtained in test A with 50 ml of a hot Magnesium hydroxide. 49.2 to 66.6 per cent of Mg(OH)2,
2 per cent w/v solution of ammonium chloride, then dissolve calculated on the dried basis and determined by the following
the precipitate in 15 ml of 3 M hydrochloric acid; the solution method. Dissolve about 100 mg, accurately weighed, in 3 ml of
gives the reactions of aluminium salts (2.3.1). dilute hydrochloric acid and dilute with water to about
200 ml. Add with stirring 1 g of ammonium chloride, 20 ml of
Tests triethanolamine, 10 ml of ammonia-ammonium chloride buffer
Arsenic (2.3.10). To 1.0 g add 15 ml of hydrochloric acid, and 0.1 ml of eriochrome black T solution and titrate with
0.1 ml of stannous chloride solution AsT and 5 ml of potassium 0.05 M disodium edetate to a blue colour. Repeat the procedure
iodide solution and allow to stand for 15 minutes. The resulting without the substance under examination. The difference
solution complies with the limit test for arsenic (10 ppm). between the titrations represents the amount of disodium
edetate required.
Heavy metals (2.3.13). Dissolve 0.33 g in 10 ml of 3 M
hydrochloric acid, filtering to get a clear solution and diluting
Mg(OH)2.
to 25 ml with water. The solution complies with the limit test
for heavy metals, Method A (60 ppm). Sulphates. 16.0 to 21.0 per cent, calculated on the dried basis
and determined by the following method. Weigh accurately
Soluble chloride. Boil 1.0 g, accurately weighed, with 50.0 ml
about 0.875 g, transfer to a 25-ml volumetric flask, dissolve in
of water for 5 minutes, cool, add water to the original volume,
10 ml of water and 5 ml of glacial acetic acid, dilute to volume
mix and filter. To 25.0 ml of the filtrate add 0.1 ml of potassium
with water and mix. Transfer 5.0 ml of this solution to a glass
chromate solution and titrate with 0.1 M silver nitrate until a
chromatographic column, 1 cm in internal diameter, prepared
persistent pink colour is obtained; Not more than 5.0 ml of
by filling with 15 ml of a strongly acidic styrene-divinylbenzene
0.1 M silver nitrate is required (3.5 per cent).
cation exchange resin (50 to 100 mesh) (such as Dowex 50W-
Soluble sulphate. Dilute 2.5 ml of the filtrate obtained in the X8 or Amberlite 120) and washing the resin with 30 ml of
test for Soluble chloride to 40 ml with water in a Nessler water. Elute the column with 15 ml of water and collect the
cylinder, add 1 ml of 3 M hydrochloric acid and 3 ml of barium eluate in a 125-ml conical flask. To the eluate add 5 ml of a
chloride solution, dilute to 50 ml, mix and allow to stand for 5.38 per cent w/v solution of magnesium acetate and 32 ml of
10 minutes; any turbidity produced is not greater than that methanol. Titrate with 0.05 M barium chloride using a 0.2 per
produced by treating 1.0 ml of 0.01 M sulphuric acid in the cent w/v solution of sodium alizarinsulphonate as the
same manner (1.9 per cent). indicator and adding about 5 ml of the titrant in the beginning
Sodium. Transfer 2.0 g, accurately weighed, to a 100-ml and continuing the titration slowly thereafter until the yellow
volumetric flask, place in an ice-bath, add 5 ml of nitric acid colour disappears and a pink tinge is visible.
and swirl to dissolve. Allow to warm to room temperature, 1 ml of 0.05 M barium chloride is equivalent to 0.004803 g of
dilute with water to volume and mix. Filter, if necessary, to SO4.
709
MAGALDRATE ORAL SUSPENSION IP 2007
Microbial contamination (2.2.9). 1 g is free from Escherichia 1 ml of 0.05 M disodium edetate is equivalent to 0.0039 g of
coli. Al(OH)3.
Loss on drying (2.4.19). 10.0 to 20.0 per cent, determined on Magnesium hydroxide. 49.2 to 66.6 per cent of the stated
0.5 g by drying in an oven at 200° for 4 hours. content of anhydrous magaldrate, determined by the following
method. Take 10.0 ml of solution A and dilute with water to
Assay. Weigh accurately about 3.0 g and transfer to a 250-ml
about 200 ml. Add with stirring 1 g of ammonium chloride,
conical flask. Add 100.0 ml of 1 M hydrochloric acid and stir
20 ml of triethanolamine, 10 ml of ammonia-ammonium
well until a clear solution is obtained. Titrate the excess acid
chloride buffer and 0.1 ml of eriochrome black T solution and
with 1 M sodium hydroxide to a pH of 3.0, determined
titrate with 0.05 M disodium edetate to a blue colour. Repeat
potentiometrically (2.4.25). Repeat the procedure without the
the procedure without the substance under examination. The
difference between the titrations represents the amount of
titrations represents the amount of hydrochloric acid required.
disodium edetate required.
1 ml of 1 M hydrochloric acid is equivalent to 0.0354 g 1 ml of 0.05 M disodium edetate is equivalent to 0.002916 g of
of Al5Mg10 (OH)31(SO4)2. Mg(OH)2.
Neutralising capacity. To an accurately weighed quantity of
the well-shaken suspension containing 0.8 g of anhydrous
magaldrate in a 250-ml beaker add water to make a total volume
Magaldrate Oral Suspension of about 70 ml, heat to 37° and stir continuously, maintaining
the temperature at 37°. Add 30.0 ml of 1 M hydrochloric acid
Magaldrate Suspension previously heated to 37° and maintain at 37° for 15 minutes,
Magaldrate Oral Suspension contains not less than 90.0 per stirring continuously. Titrate the excess acid with 1 M sodium
cent and not more than 110.0 per cent of the stated amount of hydroxide to a pH of 3.5. Not more than 12 ml of 1 M sodium
anhydrous magaldrate, Al5Mg10(OH)31(SO4)2. hydroxide is required.
Identification Microbial contamination (2.2.9). Total microbial count, not

more than 100 per ml; 0.01 ml is free from Escherichia coli.
A. Dissolve an amount of the suspension containing about Other tests. Complies with the tests stated under Oral Liquids.
0.8 g of anhydrous magaldrate in 20 ml of 3 M hydrochloric
acid, dilute with water to 50 ml, add 3 drops of methyl red Assay. To an accurately measured volume containing about
solution and heat to boiling. Add dilute ammonia solution 3.0 g of anhydrous magaldrate in a beaker add 100.0 ml of 1 M
until the colour changes to just yellow, continue boiling for hydrochloric acid and stir well until a solution is obtained.
2 minutes and filter; the filtrate gives the reactions of Titrate the excess acid with 1 M sodium hydroxide to a pH of
magnesium salts (2.3.1). 3.0, determined potentiometrically (2.4.25). Repeat the
procedure without the substance under examination. The
B. Wash the precipitate obtained in test A with 50 ml of a hot difference between the titrations represents the amount of
2 per cent w/v solution of ammonium chloride, then dissolve hydrochloric acid required.
the precipitate in 15 ml of 3 M hydrochloric acid; the solution
gives the reactions of aluminium salts (2.3.1). 1 ml of 1 M hydrochloric acid is equivalent to 0.0354 g of
Al5Mg10 (OH)31(SO4)2.
Tests Storage. Store protected from moisture.
Aluminium hydroxide. 32.1 to 45.9 per cent of the stated Labelling. The label states the strength in terms of mg of
content of anhydrous magaldrate, determined by the following anhydrous magaldrate per 5 ml.
method. To an accurately measured quantity containing about
1.0 g of magaldrate add 30 ml of dilute hydrochloric acid,
shake to dissolve, dilute to 100.0 ml with water and mix (solution Magaldrate Tablets
A). To 10.0 ml of this solution add 20 ml of water and Add with
stirring 25.0 ml of 0.05 M disodium edetate, mix and allow to Magaldrate Tablets contain not less than 90.0 per cent and
stand for 5 minutes. Add 20 ml of acetic acid-ammonium not more than 110.0 per cent of the stated amount of
acetate buffer, 60 ml ethanol (95 per cent) and 2 ml of dithizone anhydrous magaldrate, Al5Mg10(OH)31(SO4)2.
solution and titrate with 0.05 M zinc sulphate to a bright
Identification
rose-pink colour. Repeat the procedure without the substance
under examination. The difference between the titrations To a quantity of the powdered tablets containing about 2 g of
represents the amount of disodium edetate required. anhydrous magaldrate, add about 60 ml of water, shake for
710
IP 2007 HEAVY MAGNESIUM CARBONATE
3 minutes, centrifuge and discard the supernatant solution. the excess acid with 1 M sodium hydroxide to a pH of 3.5
Repeat the washing with three more quantities, each of 60 ml, determined potentiometrically (2.4.25). Not more than 12 ml of
of water. Transfer the residue to a beaker and heat on a water- 1 M sodium hydroxide is required.
bath to dryness. Disintegration (2.5.1). 2 minutes for tablets labelled to be
A. Dissolve 0.8 g of the residue in 20 ml of 3 M hydrochloric swallowed.
acid, dilute with water to 50 ml, add 3 drops of methyl red
solution and heat to boiling. Add dilute ammonia solution
until the colour changes to just yellow, continue boiling for Assay. Weigh and finely powder 20 tablets. To an accurately
2 minutes and filter; the filtrate gives the reactions of weighed quantity of the powder containing about 3.0 g of
magnesium salts (2.3.1). anhydrous magaldrate in a 200-ml volumetric flask add
100.0 ml of 1 M hydrochloric acid, shake well for 30 minutes.
B. Wash the precipitate obtained in test A with 50 ml of a hot
Dilute to volume and filter. Transfer 100.0 ml to a conical flask.
2 per cent w/v solution of ammonium chloride, then dissolve
Titrate the excess acid with 1 M sodium hydroxide to a pH of
the precipitate in 15 ml of 3 M hydrochloric acid; the solution
3.0, determined potentiometrically (2.4.25). Repeat the
gives the reactions of aluminium salts (2.3.1).
procedure without the substance under examination. The
Tests difference between the titrations represents the amount of
hydrochloric acid required.
Aluminium hydroxide. 32.1 to 45.9 per cent of the stated 1 ml of 1 M hydrochloric acid is equivalent to 0.03540 g of
content of anhydrous magaldrate, determined by the following Al5Mg10(OH)31(SO4)2.
method. Weigh and finely powder 20 tablets. To an accurately
weighed quantity of the powder containing about 1.0 g of Storage. Store protected from moisture.
anhydrous magaldrate add 30 ml of dilute hydrochloric acid, Labelling. The label states (1) the strength in terms of the
shake well to dissolve, dilute to 100.0 ml with water and mix equivalent amount of anhydrous magaldrate; (2) whether the
(solution A). To 10.0 ml of this solution add 20 ml of water and tablets are to be swallowed or chewed.
add with stirring 25.0 ml of 0.05 M disodium edetate, mix and
allow to stand for 5 minutes. Add 20 ml of acetic acid-
ammonium acetate buffer, 60 ml ethanol (95 per cent) and
2 ml of dithizone solution and titrate with 0.05 M zinc sulphate Heavy Magnesium Carbonate
to a bright rose-pink colour. Repeat the procedure without the
Heavy Magnesium Carbonate is a hydrated basic magnesium
carbonate.
titrations represents the amount of disodium edetate required.
Heavy Magnesium Carbonate contains the equivalent of not
less than 40.0 per cent and not more than 45.0 per cent of
Al(OH)3.
MgO.
Magnesium hydroxide. 49.2 to 66.6 per cent of the stated
Description. A white powder; odourless. 15 g occupies a
content of anhydrous magaldrate, determined by the following
volume of about 30 ml.
method. Take 10.0 ml of solution A and dilute with water to
about 200 ml. Add with stirring 1 g of ammonium chloride, Identification
20 ml of triethanolamine, 10 ml of ammonia-ammonium
chloride buffer and 0.1 ml of eriochrome black T solution and A. Gives reaction A of carbonates (2.3.1).
titrate with 0.05 M disodium edetate to a blue colour. Repeat B. Dissolve about 15 mg in 2 ml of 2 M nitric acid and neutralise
the procedure without the substance under examination. The with 2 M sodium hydroxide. The resulting solution gives
difference between the titrations represents the amount of reaction A of magnesium salts (2.3.1).
disodium edetate required.
1 ml of 0.05 M disodium edetate is equivalent to 0.002916 g of Tests
Mg(OH)2.
Appearance of solution. Dissolve 5.0 g in a mixture of 70 ml of
Neutralising capacity. To an accurately weighed quantity of 5 M acetic acid and 30 ml of water. Filter, if necessary, through
the powdered tablets containing 0.8 g of anhydrous magaldrate a previously ignited and weighed porcelain or silica crucible
in a 250-ml beaker add about 75 ml of water heat to 37° and stir of a suitable porosity to give a clear filtrate (solution A). Reserve
continuously, maintaining the temperature at 37°. Add 30.0 ml any residue (residue R) for the test for Substances insoluble
of 1 M hydrochloric acid previously heated to 37° and in acetic acid. Solution A is not more intensely coloured than
maintain at 37° for 15 minutes, stirring continuously. Titrate reference solution BS3 (2.4.1).
711
LIGHT MAGNESIUM CARBONATE IP 2007
Arsenic (2.3.10). Dissolve 5.0 g in 15 ml of brominated 1 ml of 0.05 M disodium edetate is equivalent to 0.002015 g of
hydrochloric acid and 45 ml of water and remove the excess MgO.
of bromine with a few drops of stannous chloride solution Storage. Store protected from moisture.
arsenic (2 ppm).
Heavy metals (2.3.13). To 20 ml of solution A add 15 ml of 7 M Light Magnesium Carbonate
hydrochloric acid and shake with 25 ml of 4-methylpentan-2-
one for 2 minutes. Separate the layers, evaporate the aqueous Light Magnesium Carbonate is a hydrated basic magnesium
layer to dryness, dissolve the residue in 1 ml of 5 M acetic carbonate.
acid and dilute to 20 ml with water. 12 ml of the resulting Light Magnesium Carbonate contains the equivalent of not
solution complies with the limit test for heavy metals, Method less than 40.0 per cent and not more than 45.0 per cent of
D (20 ppm). Use lead standard solution (1 ppm Pb) for MgO.
standard preparation.
Description. A very light, white powder. 15 g occupies a
Iron (2.3.14). Dissolve 0.1 g in 3 ml of 2 M hydrochloric acid volume of about 125 ml.
and dilute to 10 ml with water. The resulting solution complies
with the limit test for iron (400 ppm). Identification
Chlorides (2.3.12). 5.0 ml of solution A complies with the limit A. Gives reaction A of carbonates (2.3.1).
test for chlorides (0.1 per cent). B. Dissolve about 15 mg in 2 ml of 2 M nitric acid and neutralise
Sulphates (2.3.17). 0.5 ml of solution A diluted to 15 ml with with 2 M sodium hydroxide. The resulting solution gives
distilled water complies with the limit test for sulphates reaction A of magnesium salts (2.3.1).
(0.6 per cent).
Tests
(100 ppm Ca) add 1 ml of ammonium oxalate solution. After
5 M acetic acid and 30 ml of water. Filter, if necessary, through
1 minute add a mixture of 1 ml of 2 M acetic acid and 15 ml of
a previously ignited and weighed porcelain or silica crucible
a solution prepared by diluting 2.6 ml of solution A to 150 ml
of a suitable porosity to give a clear filtrate (solution A). Reserve
with distilled water and shake. After 15 minutes any
any residue (residue R) for the test for Substances insoluble
opalescence produced is not more intense than that of a
in acetic acid. Solution A is not more intensely coloured than
standard prepared in the same manner using a mixture of 10 ml
reference solution BS3 (2.4.1).
of calcium standard solution (10 ppm Ca) and 5 ml of distilled
water in place of solution A (0.75 per cent). Arsenic (2.3.10). Dissolve 5.0 g in 15 ml of brominated
Copper. Dissolve 1 g in 5 ml of hydrochloric acid and 25 ml of of bromine with a few drops of stannous chloride solution
water, boil to remove carbon dioxide, make alkaline with dilute AsT. The resulting solution complies with the limit test for
ammonia solution; no blue colour is produced. arsenic (2 ppm).
Soluble substances. Mix 2.0 g with 100 ml of water, boil for Heavy metals (2.3.13). To 20 ml of solution A add 15 ml of 7 M
5 minutes, filter whilst hot through a sintered-glass filter hydrochloric acid and shake with 25 ml of 4-methylpentan-2-
(porosity No. 3), allow to cool and dilute to 100.0 ml with one for 2 minutes. Separate the layers, evaporate the aqueous
water. Evaporate 50.0 ml of the filtrate to dryness and dry the layer to dryness, dissolve the residue in 1 ml of 5 M acetic
residue at 105°; the residue weighs not more than 10 mg (1.0 acid and dilute to 20 ml with water. 12 ml of the resulting
per cent). solution complies with the limit test for heavy metals, Method
D (20 ppm). Use lead standard solution (1 ppm Pb) to prepare
Substances insoluble in acetic acid. Residue R when washed,
the standard.
dried and ignited at 600°, weighs not more than 2.5 mg (0.05
per cent). Iron (2.3.14). Dissolve 0.1 g in 3 ml of 2 M hydrochloric acid
and dilute to 10 ml with water. The resulting solution complies
Assay. Weigh accurately about 0.15 g, dissolve in a mixture of with the limit test for iron (400 ppm).
20 ml of water and 2 ml of 2 M hydrochloric acid and add 50
ml of water, 10 ml of strong ammonia-ammonium chloride Chlorides (2.3.12). 5.0 ml of solution A complies with the limit
solution and titrate with 0.05 M disodium edetate, using 0.1 g test for chlorides (0.1 per cent).
of mordant black II mixture as indicator, until a blue colour is Calcium. To 0.2 ml of ethanolic calcium standard solution
obtained. (100 ppm Ca) add 1 ml of ammonium oxalate solution. After
712
IP 2007 MAGNESIUM CHLORIDE
1 minute add a mixture of 1 ml of 2 M acetic acid and 15 ml of Acidity or alkalinity. Dissolve 10.0 g in carbon dioxide-free
a solution prepared by diluting 2.6 ml of solution A to 150 ml water prepared from distilled water and dilute to 100 ml with
with distilled water and shake. After 15 minutes any the same solvent (solution A). To 5 ml of solution A add
opalescence produced is not more intense than that of a 0.05 ml of phenol red solution. Not more than 0.3 ml of either
standard prepared in the same manner using a mixture of 10 ml 0.01 M hydrochloric acid or 0.01 M sodium hydroxide is
of calcium standard solution (10 ppm Ca) and 5 ml of distilled required to change the colour of the solution.
water in place of solution A (0.75 per cent). Arsenic (2.3.10). Dissolve 5.0 g in 50 ml of water and add 10 ml
Copper. Dissolve 1 g in 5 ml of hydrochloric acid and 25 ml of of stannated hydrochloric acid. The resulting solution
water, boil to remove carbon dioxide, make alkaline with dilute complies with the limit test for arsenic (2 ppm).
ammonia solution; no blue colour is produced. Heavy metals (2.3.13). Dissolve 1.0 g in 20 ml of water, adding
Soluble substances. Mix 2.0 g with 100 ml of water, boil for 1 g of ammonium chloride and sufficient water to produce 25
5 minutes, filter whilst hot through a sintered-glass filter ml. The resulting solution complies with the limit test for heavy
(porosity No. 3), allow to cool and dilute to 100.0 ml with metals, Method A (20 ppm).
water. Evaporate 50.0 ml of the filtrate to dryness and dry the Iron (2.3.14). 4.0 g complies with the limit test for iron (10
residue at 105°; the residue weighs not more than 10 mg (1.0 ppm).
per cent).
Calcium. To 25 ml of solution A add sufficient water to produce
Substances insoluble in acetic acid. Residue R when washed, 50 ml and determine by Method B for flame photometry (2.4.4),
dried and ignited at 600°, weighs not more than 2.5 mg (0.05 or by Method B for atomic absorption spectrophotometry
per cent). (2.4.2), measuring at 423 nm and using calcium solution FP,
Sulphates (2.3.17). 1 ml of solution A diluted to 15 ml with or calcium solution AAS, as appropriate, diluted if necessary,
distilled water complies with the limit test for sulphates (0.3 with 0.1 M hydrochloric acid for the standard solution (0.1
per cent). per cent).
Assay. Weigh accurately about 0.15 g, dissolve in a mixture of Sulphates (2.3.17). 15 ml of solution A complies with the limit
20 ml of water and 2 ml of 2 M hydrochloric acid and add 50 test for sulphates (100 ppm).
ml of water, 10 ml of strong ammonia-ammonium chloride Assay. Weigh accurately about 0.3 g, dissolve in 30 ml of
solution and titrate with 0.05 M disodium edetate, using 0.1 g water, add 10 ml of strong ammonia-ammonium chloride
of mordant black II mixture as indicator, until a blue colour is solution and titrate with 0.05 M disodium edetate using 0.1 g
obtained. of mordant black II mixture as indicator.
1 ml of 0.05 M disodium edetate is equivalent to 0.002015 g of 1 ml of 0.05 M disodium edetate is equivalent to 0.01017 g of
MgO. MgCl2,6H2O.
Storage. Store protected from moisture Magnesium Chloride intended for use in the preparation of
dialysis solutions complies with the following additional
requirement.
Aluminium. Dissolve 4.0 g in 100 ml of water and add 10 ml of
Magnesium Chloride acetate buffer pH 6.0. Extract with successive quantities of
MgCl2,6H2O Mol. Wt. 203.3 20, 20 and 10 ml of a 0.5 per cent w/v solution of 8-
hydroxyquinoline in chloroform and dilute the combined
Magnesium Chloride contains not less than 98.0 per cent extracts to 50 ml with chloroform. Use as the standard solution
and not more than 101.0 per cent of MgCl2,6H2O. a mixture of 2 ml of aluminium standard solution (2 ppm Al),
Description. Colourless crystals; hygroscopic. 10 ml of acetate buffer pH 6.0 and 98 ml of water treated in the
same manner and as the blank solution a mixture of 10 ml of
Identification acetate buffer pH 6.0 and 100 ml of water treated in the same
manner. Measure the fluorescence of the test solution and the
A. Gives reaction A of chlorides (2.3.1).
standard solution (2.4.5), using an excitation wavelength of
B. Gives reaction A of magnesium salts (2.3.1). about 392 nm and emission wavelength of about 518 nm, and
setting the instrument to zero with the blank solution in each
Tests case. The fluorescence of the test solution is not greater than
Appearance of solution. Solution A is clear (2.4.1), and that of the standard solution (1 ppm).
colourless (2.4.1). Storage. Store protected from moisture.
713
MAGNESIUM HYDROXIDE IP 2007
Magnesium Hydroxide opalescence produced is not more intense than that of a

Mg (OH)2 Mol. Wt. 58.3 of calcium standard solution (10 ppm Ca) and 5 ml of distilled
Magnesium Hydroxide contains not less than 95.0 per cent water in place of solution A (1.5 per cent).
and not more than 100.5 per cent of Mg(OH)2. Soluble substances. Mix 2.0 g with 100 ml of water, boil for 5
Description. A bulky white powder. minutes, filter whilst hot through a sintered-glass filter
(porosity No. 3), allow to cool and dilute to 100.0 ml with
Identification water. Evaporate 50.0 ml of the filtrate to dryness and dry the
residue at 105°; the residue weighs not more than 10 mg (1.0
Dissolve about 15 mg in 2 ml of 2 M nitric acid and neutralise per cent).
with 2 M sodium hydroxide. The resulting solution gives
reaction A of magnesium salts (2.3.1). Substances insoluble in acetic acid. Residue R when washed,
dried and ignited at 600°, weighs not more than 5.0 mg (0.1 per
Tests cent).
Appearance of solution. Dissolve 5.0 g in a mixture 50 ml of 5 Loss on ignition (2.4.20). 30.0 to 32.5 per cent, determined on
M acetic acid and 50 ml of distilled water; not more than a 0.5 g by igniting at 900° increasing the heat gradually.
slight effervescence is produced. Boil for 2 minutes, cool and Assay. Weigh accurately about 0.1 g, dissolve in 20 ml of 2 M
dilute to 100 ml with 2 M acetic acid. Filter, if necessary, hydrochloric acid and dilute to 100.0 ml with water. To 50.0 ml
through a previously ignited and weighed porcelain or silica of this solution add 5 ml of strong ammonia-ammonium
crucible of a suitable porosity to give a clear filtrate (solution chloride solution and titrate with 0.05 M disodium edetate,
A). Reserve any residue (residue R) for the test for Substances using about 50 mg of mordant black II mixture as indicator.
insoluble in acetic acid. Solution A is not more intensely
Mg(OH)2.
arsenic (4 ppm).
Magnesium Hydroxide Oral
Heavy metals (2.3.13). To 20 ml of solution A add 15 ml of 7 M
hydrochloric acid and shake with 25 ml of 4-methylpentan-2- Suspension
one for 2 minutes. Separate the layers, evaporate the aqueous
Magnesium Hydroxide Mixture; Milk of Magnesia;
layer to dryness, dissolve the residue in 1 ml of 5 M acetic
Cream of Magnesia
acid and dilute to 20 ml with water. 12 ml of the resulting
solution complies with the limit test for heavy metals, Method Magnesium Hydroxide Oral Suspension is an aqueous
D (30 ppm). Use lead standard solution (2 ppm Pb) to prepare suspension of hydrated magnesium oxide. It may be prepared
the standard. from a suitable grade of Light Magnesium Oxide.
Iron (2.3.14). Dissolve 0.2 g in 7 ml of 2 M hydrochloric acid Magnesium Hydroxide Oral Suspension contains not less than
and dilute to 20 ml with water. 5 ml of the resulting solution 7.0 per cent and not more than 8.5 per cent w/w of hydrated
complies with the limit test for iron (0.08 per cent). magnesium oxide, calculated as Mg(OH)2.
Chlorides (2.3.12). 5.0 ml of solution A diluted to 15 ml with Description. A white, uniform suspension, which does not
distilled water complies with the limit test for chlorides (0.1 separate readily on standing.
per cent).
Identification
Sulphates (2.3.17). 0.6 ml of solution A diluted to 15 ml with
distilled water complies with the limit test for sulphates (0.5 A solution of 1 ml in 2 ml of dilute hydrochloric acid gives the
per cent). reactions of magnesium salts (2.3.1).
Calcium. To 0.2 ml of ethanolic calcium standard solution Tests
(100 ppm Ca) add 1 ml of ammonium oxalate solution. After
1 minute add a mixture of 1 ml of 2 M acetic acid and 15 ml of Soluble alkalis. Filter about 25 ml and discard the first 10 ml of
a solution prepared by diluting 1.3 ml of solution A to 150 ml the filtrate. Dilute 5 ml of the filtrate with 40 ml of water, add
with distilled water and shake. After 15 minutes any 0.05 ml of methyl red solution and titrate with 0.05 M sulphuric
714
IP 2007 HEAVY MAGNESIUM OXIDE
acid to a persistent pink colour. Not more than 1.0 ml of Tests

0.05 M sulphuric acid is required.
Soluble salts. To 5 ml of the clear filtrate obtained in the test 5 M acetic acid and 30 ml of water. Filter, if necessary, through
for Soluble alkalis add 0.15 ml of sulphuric acid, evaporate to a previously ignited and weighed porcelain or silica crucible
dryness on a water-bath and then ignite gently to constant of a suitable porosity to give a clear filtrate (solution A). Reserve
weight; the residue does not weigh more than 12 mg. any residue (residue R). Solution A is not more intensely
Heavy metals (2.3.13). Dissolve 12.5 g in 10 ml of hydrochloric coloured than reference solution BS3 (2.4.1).
acid and 20 ml of water, add 0.5 ml of nitric acid, boil to Arsenic (2.3.10). Dissolve 2.5 g in 15 ml of brominated
remove any carbon dioxide and filter. To the cooled filtrate hydrochloric acid, add 45 ml of water and remove the excess
add 2 g of ammonium chloride and 2 g of ammonium of bromine with a few drops of stannous chloride solution
thiocyanate and extract with two successive quantities, each AsT. The resulting solution complies with the limit test for
of 10 ml, of ether. To the aqueous layer add 2 g of citric acid arsenic (4 ppm).
and sufficient water to produce 50 ml. 12 ml of the solution Heavy metals (2.3.13). 20 ml of solution A complies with the
complies with the limit test for heavy metals, Method D limit test for heavy metals, Method D (30 ppm).
(4 ppm). Use lead standard solution (1 ppm Pb) to prepare
Iron (2.3.14). Dissolve 0.2 g in 7 ml of 2 M hydrochloric acid
the standard.
and dilute to 20 ml with water. 5 ml of the resulting solution
Sulphates (2.3.17). Dissolve 2.5 ml in 20 ml of hydrochloric complies with the limit test for iron (800 ppm).
acid and dilute to 500 ml with water. 15 ml of the resulting
Chlorides (2.3.12). 5.0 ml of solution A complies with the limit
solution, filtered if necessary, complies with the limit test for
test for chlorides (0.1 per cent).
sulphates (0.2 per cent).
Sulphates (2.3.17). 0.3 ml of solution A complies with the limit
Microbial contamination (2.2.9). Total microbial count, not test for sulphates (1.0 per cent).
more than 100 per ml; 1 ml is free from Escherichia coli.
Other tests. Complies with the tests stated under Oral Liquids. (100 ppm Ca) add 1 ml of ammonium oxalate solution. After
Assay. Weigh accurately about 10.0 g, mix with 50 ml of water, 1 minute add a mixture of 1 ml of 2 M acetic acid and 15 ml of
add 50.0 ml of 0.5 M sulphuric acid and titrate the excess of a solution prepared by diluting 1.3 ml of solution A to 150 ml
acid with 1 M sodium hydroxide using methyl orange solution with distilled water and shake. After 15 minutes any
as indicator. opalescence produced is not more intense than that of a
1 ml of 0.5 M sulphuric acid is equivalent to 0.02916 g of of calcium standard solution (10 ppm Ca) and 5 ml of distilled
hydrated magnesium oxide calculated as Mg(OH)2. water in place of solution A (1.5 per cent).
Storage. Store protected from moisture. Do not keep in a Soluble substances. Mix 2.0 g with 100 ml of water, boil for 5
refrigerator. minutes, filter whilst hot through a sintered-glass filter
(porosity No. 3), allow to cool and dilute to 100.0 ml with
water. Evaporate 50.0 ml of the filtrate to dryness and dry the
residue at 105°. The residue weighs not more than 20 mg (2.0
Heavy Magnesium Oxide per cent).
Substances insoluble in acetic acid. Residue R when washed
Heavy Magnesia
with water, dried and ignited at 600° , weighs not more than 5
MgO Mol. Wt. 40.3 mg (0.1 per cent).
Heavy Magnesium Oxide contains not less than 98.0 per Loss on ignition (2.4.20). Not more than 8.0 per cent,
cent and not more than 100.5 per cent of MgO, calculated determined on 0.5 g when ignited at 900°.
on the ignited basis. Assay. Weigh accurately about 0.35 g, dissolve in 10 ml of 2 M
Description. A fine, white powder. 15 g occupies a volume of hydrochloric acid and dilute with water to 100.0 ml. To 10.0
about 30 ml. ml add 5 ml of strong ammonia-ammonium chloride solution
and titrate with 0.05 M disodium edetate using about 50 mg
Identification of mordant black II mixture as indicator.
Dissolve about 15 mg in 2 ml of 2 M nitric acid and neutralise 1 ml of 0.05 M disodium edetate is equivalent to 0.002015 g of
with 2 M sodium hydroxide. The resulting solution gives MgO.
reaction A of magnesium salts (2.3.1). Storage. Store protected from moisture.
715
LIGHT MAGNESIUM OXIDE IP 2007
Light Magnesium Oxide opalescence produced is not more intense than that of a
Light Magnesia of calcium standard solution (10 ppm Ca) and 5 ml of distilled
MgO Mol. Wt. 40.3 water in place of solution A (1.5 per cent).
Light Magnesium Oxide contains not less than 98.0 per cent Soluble substances. Mix 2.0 g with 100 ml of water, boil for
and not more than 100.5 per cent of MgO, calculated on the 5 minutes, filter whilst hot through a sintered-glass filter
ignited basis. (porosity No. 3), allow to cool and dilute to 100.0 ml with
water. Evaporate 50.0 ml of the filtrate to dryness and dry the
Description. A very fine, light, white powder. 15 g occupies a residue at 105°. The residue weighs not more than 20 mg
volume of about 150 ml. (2.0 per cent).
Identification Substances insoluble in acetic acid. Residue R when washed

with water, dried and ignited at 600°, weighs not more than
Dissolve about 15 mg in 2 ml of 2 M nitric acid and neutralise 5 mg (0.1 per cent).
with 2 M sodium hydroxide. The resulting solution gives
Loss on ignition (2.4.20). Not more than 8.0 per cent,
reaction A of magnesium salts (2.3.1).
determined on 0.5 g when ignited at 900°.
Tests Assay. Weigh accurately about 0.35 g, dissolve in 10 ml of 2 M
hydrochloric acid and dilute with water to 100.0 ml. To
Appearance of solution. Dissolve 5.0 g in a mixture of 70 ml of 10.0 ml add 5 ml of strong ammonia-ammonium chloride
5 M acetic acid and 30 ml of water. Filter, if necessary, through solution and titrate with 0.05 M disodium edetate using about
a previously ignited and weighed porcelain or silica crucible 50 mg of mordant black II mixture as indicator.
of a suitable porosity to give a clear filtrate (solution A). Reserve
any residue (residue R) for the test for Substances insoluble 1 ml of 0.05 M disodium edetate is equivalent to 0.002015 g of
in acetic acid. Solution A is not more intensely coloured than MgO.
reference solution BS2 (2.3.1). Storage. Store protected from moisture.
AsT. The resulting solution complies with the limit test for Magnesium Stearate
arsenic (4 ppm).
Magnesium Stearate consists mainly of magnesium stearate
Heavy metals (2.3.13). To 20 ml of solution A add 15 ml of 7 M (C17H35CO2)2Mg with variable proportions of magnesium
hydrochloric acid and shake with 25 ml of 4-methylpentan-2- palmitate, (C 15 H 31 CO 2 ) 2 Mg and magnesium oleate,
one for 2 minutes. Separate the layers, evaporate the aqueous (C17H33CO2)2Mg.
layer to dryness, dissolve the residue in 1 ml of 5 M acetic
acid and dilute to 20 ml with water. 12 ml of the resulting Magnesium Stearate contains not less than 3.8 per cent and
solution complies with the limit test for heavy metals, Method not more than 5.0 per cent of Mg, calculated on the dried
D (30 ppm). Use 7.5 ml of lead standard solution (2 ppm Pb) basis.
for preparing the standard. Description. A very fine, light, white powder; odourless or
Iron (2.3.14). Dissolve 40 mg in 5 ml of 2 M hydrochloric acid with a very faint odour of stearic acid; unctuous and free from
and dilute to 10 ml with water. The resulting solution complies grittiness.
with the limit test for iron (0.1 per cent).
Identification
Chlorides (2.3.12). 4.0 ml of solution A complies with the limit
test for chlorides (0.125 per cent). A. To 5.0 g add 50 ml of ether, 20 ml of 2 M nitric acid and 20
ml of distilled water and heat under a reflux condenser until
Sulphates (2.3.17). 0.3 ml of solution A complies with the limit
dissolution is complete. Allow to cool, separate the aqueous
test for sulphates (1.0 per cent).
layer and shake the ether layer with two quantities, each of 4
Calcium. To 0.2 ml of ethanolic calcium standard solution ml, of distilled water. Combine the aqueous layers, wash with
(100 ppm Ca) add 1 ml of ammonium oxalate solution. After 15 ml of ether and dilute to 50 ml with distilled water (solution
1 minute add a mixture of 1 ml of 2 M acetic acid and 15 ml of A). Evaporate the ether layer to dryness and dry the residue
a solution prepared by diluting 1.3 ml of solution A to 150 ml at 105°. The freezing point of the residue is not lower than 53°
with distilled water and shake. After 15 minutes any (2.4.11).
716
IP 2007 MAGNESIUM SULPHATE
B. 1 ml of solution A obtained in test A gives reaction A of Sulphates (2.3.17). Dilute 5.0 ml of solution A to 50.0 ml with
magnesium salts (2.3.1). water. 2.5 ml of this solution diluted to 15 ml with water
complies with the limit test for sulphates (0.6 per cent)
Tests
Appearance of solution. Solution A is not more intensely on 1.0 g by drying in an oven at 105°.
Assay. Weigh accurately about 0.75 g, add 50 ml of a mixture
Appearance of solution of the fatty acids. Dissolve 0.5 g of the of equal volumes of 1-butanol and ethanol, 5 ml of strong
residue obtained in the preparation of solution A in 10 ml of ammonia solution, 3 ml of ammonia buffer pH 10.0, 30.0 ml of
chloroform. The solution is clear (2.4.1), and not more intensely 0.1 M disodium edetate and 15 mg of mordant black II mixture,
coloured than reference solution YS5 (2.4.1). heat to 45° to 50° and titrate with 0.1 M zinc sulphate until the
Acidity or alkalinity. Mix 1.0 g with 20 ml of carbon dioxide- colour changes from blue to violet. Repeat the operation
free water, boil for 1 minute, shaking continuously, cool and without the substance under examination. The difference
filter. To 10 ml of filtrate add 0.05 ml of bromothymol blue between the titrations represents the amount of disodium
solution. Not more than 0.05 ml of 0.1 M hydrochloric acid or edetate required.
the solution.
Mg.
Acid value of the fatty acids. 195 to 210, determined on 0.2 g of
the residue obtained in the preparation of solution A, dissolved
in 25 ml of the prescribed mixture of solvents (2.3.23).
Free stearic acid. Not more than 3 per cent, determined by the Magnesium Sulphate
following method. Weigh accurately about 1.0 g into a
stoppered flask, add 50 ml of chloroform, stopper the flask Epsom Salts
and shake well. Filter into a beaker through two thicknesses MgSO4,7H2O Mol. Wt. 246.5
of filter paper taking care to avoid evaporation of the solvent.
Wash the filter with 10 ml of chloroform and collect the Magnesium Sulphate contains not less than 99.0 per cent
washings in the beaker. Evaporate the chloroform on a water- and not more than 100.5 per cent of MgSO4, calculated on
bath in a current of air. Dissolve the residue in about 10 ml of the dried basis.
ethanol (95 per cent) previously neutralised to Description. Colourless crystals or a white, crystalline powder.
phenolphthalein solution and titrate with 0.1 M sodium
hydroxide using phenolphthalein solution as indicator. Identification
A. Gives the reactions of sulphates (2.3.1).
stearic acid.
Zinc stearate. Heat 5.0 g with shaking with a mixture of 50 ml B. Gives reaction A of magnesium salts (2.3.1).
of water and 50 ml of dilute sulphuric acid until the fatty
acids separate as an oily layer. Cool, filter the aqueous layer
Tests
and wash the residue with two successive quantities, each of Appearance of solution. Dissolve 5.0 g in sufficient carbon
5 ml, of hot water, combine the filtrate and the washings and dioxide-free water to produce 50 ml (solution A). Solution A
dilute to 100 ml with water. To 5 ml of the resulting solution is clear (2.4.1), and colourless (2.4.1).
add 0.5 ml of ammonium mercurithiocyanate solution and
0.05 ml of copper sulphate solution. Scratch the walls of the Acidity or alkalinity. To 10 ml of solution A add 0.05 ml of
container with a glass rod and allow to stand for 15 minutes; phenol red solution. Not more than 0.2 ml of either 0.01 M
no violet precipitate is formed. hydrochloric acid or 0.01 M sodium hydroxide is required to
change the colour of the solution.
Heavy metals (2.3.13). Heat 5.0 g with 40 ml of 2 M acetic acid
and allow to cool. Filter, wash the residue with two quantities, Arsenic (2.3.10). Dissolve 5.0 g in 50 ml of water and add 10 ml
each of 5 ml, of warm water and dilute to 100 ml with water. of stannated hydrochloric acid. The resulting solution
12 ml of the resulting solution complies with the limit test for complies with the limit test for arsenic (2 ppm).
heavy metals, Method D (20 ppm). Use 1.0 ml of lead standard Heavy metals (2.3.13). Dissolve 2.0 g in 10 ml of water, 2 ml of
solution (10 ppm Pb) to prepare the standard. dilute acetic acid and sufficient water to produce 25 ml. The
Chlorides (2.3.12). 10.0 ml of solution A diluted to 15 ml solution complies with the limit test for heavy metals, Method
complies with the limit test for chlorides (250 ppm). A (10 ppm).
717
MAGNESIUM TRISILICATE IP 2007
Iron (2.3.14). 2.0 ml of solution A diluted to 20 ml with water Heavy metals (2.3.13). Neutralise 7.5 ml of solution A with
complies with the limit test for iron (200 ppm). dilute ammonia solution using metanil yellow solution as
Chlorides (2.3.12). 8.0 ml of solution A complies with the limit external indicator, dilute to 15 ml with water and filter if
test for chlorides (300 ppm). necessary. 12 ml of the resulting solution complies with the
limit test for heavy metals, Method D (40 ppm).
0.5 g by drying in an oven at 110° to 120° for 1 hour and then Chlorides (2.3.12). 0.5 ml of solution A diluted to 15 ml with
at 400°. distilled water complies with the limit test for chlorides (500
ppm).
water, add 10 ml of strong ammonia-ammonium chloride Sulphates (2.3.17). 0.3 ml of solution A diluted to 15 ml with
solution and titrate with 0.05 M disodium edetate, using 0.1 g distilled water complies with the limit test for sulphates (0.5
of mordant black II mixture as indicator, until a blue colour is per cent).
obtained.
Acid absorption. Not less than 100 ml of 0.1 M hydrochloric
1 ml of 0.05 M disodium edetate is equivalent to 0.00602 g of acid per g, determined by the following method. Suspend 0.25
MgSO4. g in 100.0 ml of 0.1 M hydrochloric acid, allow to stand in a
Storage. Store protected from moisture. water-bath at 37° ± 0.5° for 2 hours, shaking frequently and
allow to cool. Add 0.1 ml of bromophenol blue solution to
20.0 ml of the supernatant liquid and titrate with 0.1 M sodium
hydroxide until a blue colour is produced.
Magnesium Trisilicate
Water-soluble salts. In a tared platinum dish evaporate to
Magnesium Trisilicate is a hydrated magnesium silicate of the dryness on a water-bath 20 ml of the clear liquid obtained in
approximate composition 2MgO,3SiO2,xH2O. the test for Alkalinity and ignite the residue to constant weight
Magnesium Trisilicate contains not less than the equivalent at 900°. The residue weighs not more than 30 mg.
of 29.0 per cent of MgO and not less than the equivalent of Loss on ignition (2.4.20). 17.0 to 34.0 per cent, determined on
65.0 per cent of SiO2, both calculated on the ignited basis. 0.5 g when ignited in a platinum crucible at 900°.
Description. A fine, white or nearly white powder, free from Assay. For MgO — Weigh accurately about 1.0 g, add 35 ml
grittiness; slightly hygroscopic.
of hydrochloric acid and 50 ml of water and allow to stand for
Identification 15 minutes on a water-bath. Allow to cool, filter, wash the
residue with water and dilute the combined filtrate and
A. To 2.0 g add a mixture of 4 ml of nitric acid and 4 ml of washings to 250.0 ml with water. Neutralise 50.0 ml with about
distilled water and heat to boiling, shaking frequently. Add 8 ml of 10 M sodium hydroxide, add 10 ml of ammonia buffer
12 ml of distilled water, allow to cool, filter or centrifuge to pH 10.0, 50 mg of mordant black II mixture, heat to 40° and
obtain a clear solution and dilute the filtrate to 20 ml with titrate with 0.05 M disodium edetate until the colour changes
distilled water (solution A). 1 ml of solution A, after to a deep blue.
neutralisation with 2 M sodium hydroxide, gives reaction A of
magnesium salts (2.3.1). 1 ml of 0.05 M disodium edetate is equivalent to 0.002015 g of
MgO.
B. 0.25 g gives the reaction of silicates (2.3.1).
For SiO2 — Weigh accurately about 0.7 g, add 10 ml of 1 M
Tests sulphuric acid and 10 ml of water and heat on a water-bath for
1.5 hours, shaking frequently and replacing the evaporated
Alkalinity. In a tared 200-ml conical flask on a water-bath heat
water. Allow to cool, decant onto an ashless filter paper (7 cm
10 g with 100 g of water for 30 minutes, allow to cool and
in diameter), wash the precipitate by decantation with three
restore the initial weight with water. Allow to stand and filter
quantities, each of 5 ml, of hot water, transfer it to the filter
or centrifuge until a clear liquid is obtained. To 10 ml of the
paper and wash it with hot water until 1 ml of the filtrate remains
clear liquid add 0.1 ml of phenolphthalein solution. Not more
clear on the addition of 2 ml of barium chloride solution and
than 1.0 ml of 0.1 M hydrochloric acid is required to change
the colour of the solution. 0.05 ml of 2 M hydrochloric acid. Ignite the filter paper and its
contents in a tared platinum crucible at 900° to constant weight;
Arsenic (2.3.10). Disperse 2.5 g in 50 ml of water and add 10 ml the residue is SiO2.
of stannated hydrochloric acid AsT. The resulting solution
complies with the limit test for arsenic (4 ppm). Storage. Store protected from moisture.
718
IP 2007 MANNITOL
Mannitol water previously heated at 30° and shaking continuously for

15 to 30 minutes without further heating. Dilute the resulting
D-Mannitol clear solution to 25.0 ml with water.
Arsenic (2.3.10). Dissolve 5.0 g in 50 ml of water and add 10 ml
CH2OH of stannated hydrochloric acid. The resulting solution
HO H complies with the limit test for arsenic (2 ppm).
HO H Chlorides (2.3.12). A solution of 5.0 g in 10 ml of water
H OH complies with the limit test for chlorides (50 ppm).
H OH Sulphates (2.3.17). A solution of 1.5 g in 10 ml of water complies
CH2OH with the limit test for sulphates (100 ppm).
Reducing sugars. Dissolve 5.0 g in 25 ml of water with the aid
C6H14O6 Mol. Wt. 182.2 of gentle heat. Cool and add 20 ml of cupri-citric solution and
a few glass beads. Heat so that boiling begins 4 minutes later
Mannitol is D-mannitol, a hexahydric alcohol related to
and continue to boil for 3 minutes. Cool rapidly and add 100 ml
mannose.
of a 2.4 per cent v/v solution of glacial acetic acid and 20.0 ml
Mannitol contains not less than 98.0 per cent and not more of 0.025 M iodine. With continuous shaking add 25 ml of a
than 102.0 per cent of C6H14O6, calculated on the dried basis. mixture of 6 volumes of hydrochloric acid and 94 volumes of
Description. A white, crystalline powder or free-flowing water and, when the precipitate has dissolved, titrate the
granules. excess of iodine with 0.05 M sodium thiosulphate using 1 ml
of starch solution, added towards the end of titration, as
Identification indicator. Not less than 12.8 ml of 0.05 M sodium thiosulphate
is required.
A. To 1 ml of a saturated solution add 0.5 ml of ferric chloride
test solution followed by 0.25 ml of sodium hydroxide solution Sorbitol. Determine by thin-layer chromatography (2.4.17),
and shake well; a clear solution is obtained which remains coating the plate with a uniform 0.75-mm layer of the following
clear on the further addition of sodium hydroxide solution. mixture. Mix 0.1 g of carbomer with 110 ml of water and allow
to stand, with gentle stirring, for 1 hour. Adjust to pH 7 by the
B. Dissolve 5 g in sufficient carbon dioxide-free water gradual addition, with continuous shaking, of 2 M sodium
prepared from distilled water to produce 50 ml (solution A). hydroxide and add 30 g of silica gel H. Heat the plate at 110°
Add 0.3 ml of solution A to 3 ml of a cooled mixture prepared for 1 hour, allow to cool and use immediately.
by adding 6 ml of sulphuric acid to 3 ml of a freshly prepared
10 per cent w/v solution of catechol while cooling in ice. Heat Mobile phase. A mixture of 85 volumes of 2-propanol and 15
gently over a naked flame for about 30 seconds; a pink colour volumes of a 0.2 per cent w/v solution of boric acid.
is produced. Test solution. Shake 0.5 g of the substance under examination,
C. Melting range 165° to 170° (2.4.21). in fine powder, with 10 ml of ethanol (95 per cent) for 30
minutes and filter.
Tests Reference solution. A 0.1 per cent w/v solution of sorbitol RS
in ethanol (95 per cent).
Appearance of solution. Solution A is clear (2.4.1), and
colourless (2.4.1). Apply to the plate 2 µl of each solution. After development,
Acidity or alkalinity. To 5 ml of solution A add 5 ml of carbon dry the plate at 100° to 105° for 15 minutes, allow to cool,
dioxide-free water and 0.05 ml of dilute phenolphthalein spray with a 0.5 per cent w/v solution of potassium
solution; not more than 0.2 ml of 0.01 M sodium hydroxide is permanganate in 1 M sodium hydroxide and heat at 100° for
required to change the colour of the solution to pink. To a 2 minutes. Any spot corresponding to sorbitol in the
further 5 ml of solution A add 5 ml of carbon dioxide-free chromatogram obtained with the test solution is not more
water and 0.05 ml of methyl red solution. Not more than 0.3 ml intense than the spot in the chromatogram obtained with the
of 0.01 M hydrochloric acid is required to change the colour reference solution.
of the solution to red. Sulphated ash (2.3.18). Not more than 0.1 per cent, determined
Specific optical rotation (2.4.22). +23.0° to +25.0°, determined on 2.0 g.
in a solution prepared by dissolving 2.0 g of the substance Loss on drying (2.4.19). Not more than 0.5 per cent, determined
under examination and 2.6 g of sodium tetraborate in 20 ml of on 1.0 g by drying in an oven at 105°.
719
MANNITOL INJECTION IP 2007
Assay. Determine by liquid chromatography (2.4.14). B. To the residue obtained in test A add 0.5 ml of ferric chloride
test solution followed by 0.25 ml of sodium hydroxide solution
and shake well; a clear solution is obtained which remains
examination in 25 ml of water and dilute to 100.0 ml with water.
clear on the further addition of sodium hydroxide solution.
Reference solution. Dissolve 0.5 g of mannitol RS in 2.5 ml of
water and dilute to 10.0 ml with water. C. Dissolve 0.5 g of the residue obtained in test A in sufficient
carbon dioxide-free water prepared from distilled water to
Chromatographic system produce 5 ml (solution A). Add 0.3 ml of solution A to 3 ml of
– a stainless steel column 30 cm x 7.8 mm packed with a cooled mixture prepared by adding 6 ml of sulphuric acid to
strong cation-exchange resin (calcium form) (9 µm), 3 ml of a freshly prepared 10 per cent w/v solution of catechol
– column temperature. 85 ± 1°, while cooling in ice. Heat gently over a naked flame for about
– mobile phase: degassed water, 30 seconds; a pink colour is produced.
– refractometer at constant temperature, Tests
pH (2.4.24). 4.5 to 7.0, determined in a solution containing not
Inject the test solution and the reference solution. Continue more than 10.0 per cent w/v solution of Mannitol, diluted if
the chromatography for twice the retention time of mannitol. necessary with water and to which 0.3 ml of a saturated
Calculate the content of C6H14O6. solution of potassium chloride has been added for each 100
ml of solution.
Mannitol intended for use in the manufacture of parenteral
preparations without a further appropriate procedure for Particulate contamination (2.5.9). When supplied in a
removal of bacterial endotoxins complies with the following container with a nominal content of 100 ml or more, complies
additional requirement. with the limit test for particulate contamination.
Bacterial endotoxins (2.2.3). Not more than 4 Endotoxin Unit Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit
per g for parenteral preparation having a concentration of 100 per ml of a solution containing not more than 10 per cent w/v
g per litre or less of mannitol, and less than 2.5 Endotoxin per of Mannitol. For solutions of higher strength, dilute the
g for parenteral preparations having a concentration of more injection under examination with water BET so that the final
than 100 g per litre of mannitol. solution contains 10 per cent w/v of Mannitol.
Labelling. The label states where applicable, the maximum Other tests. Complies with the tests stated under Parenteral
concentration of bacterial endotoxins; whether or not the Preparations (Intravenous Infusions).
substance is suitable for use in the manufacture of parenteral
Assay. Dilute an accurately measured volume containing about
preparations.
0.4 g of Mannitol to 100.0 ml with water, transfer 10.0 ml to a
Storage. Store protected from moisture. stoppered flask, add 20.0 ml of 0.1 M sodium periodate and 2
ml of 1 M sulphuric acid and heat on a water-bath for 15
minutes. Cool, add 3 g of sodium bicarbonate, in small
quantities, and 25.0 ml of 0.1 M sodium arsenite, mix, add 5 ml
Mannitol Injection of a 20 per cent w/v solution of potassium iodide and allow to
stand for 15 minutes. Titrate with 0.05 M iodine until the first
Mannitol Intravenous Infusion trace of yellow colour appears. Repeat the operation without
Mannitol Injection is a sterile solution of Mannitol in Water the substance under examination. The difference between the
for Injections. titrations represents the amount of iodine required.
Mannitol Injection contains not less than 95.0 per cent and 1 ml of 0.05 M iodine is equivalent to 0.001822 g of C6H14O6.
not more than 105.0 per cent of the stated amount of mannitol, Storage. Store at temperatures between 20° and 30°. Exposure
C6H14O6. to lower temperatures may cause the deposition of crystals,
Description. A colourless or almost colourless clear solution. which should be dissolved by warming before use.
Labelling. The label states (1) the strength as a percentage
Identification
w/v of Mannitol; (2) that the injection should not be used if
A. Evaporate to dryness on a water-bath a volume containing it contains visible solid particles that do not dissolve
2 g of Mannitol. The residue melts at 165° to 170° (2.4.21). on warming.
720
IP 2007 MEBENDAZOLE TABLETS
Mebendazole Any secondary spot in the chromatogram obtained with the

H O
OCH3 Heavy metals (2.3.13). 1.0 g complies with the limit test for
N
NH
N Sulphated ash (2.3.18). Not more than 0.1 per cent.
C16H13N3O3 Mol. Wt. 295.3 Assay. Weigh accurately about 0.25 g, dissolve in 3 ml of
Mebendazole is methyl N-(5-benzoyl-1H-benzimidazol-2- anhydrous formic acid and 30 ml of anhydrous glacial acetic
yl)carbamate. acid. Titrate with 0.1 M perchloric acid, determining the end-
point potentiometrically (2.4.25). Carry out a blank titration.
Mebendazole contains not less than 98.0 per cent and not
more than 102.0 per cent of C16H13N3O3, calculated on the 1 ml of 0.1 M perchloric acid is equivalent to 0.02953 g of
dried basis. C16H13N3O3.
Description. A white to slightly yellow, amorphous powder; Storage. Store protected from light and moisture.
almost odourless.
Identification
Mebendazole Tablets
Compare the spectrum with that obtained with mebendazole Mebendazole Tablets contain not less than 92.5 per cent and
RS or with the reference spectrum of mebendazole. not more than 107.5 per cent of the stated amount of
B. In the test for Related substances, the principal spot in the mebendazole, C16H13N3O3.
that in the chromatogram obtained with reference solution (a). Identification
C. To about 10 mg add 5 ml of ethanol (95 per cent), 1 ml of A. Shake a quantity of the powdered tablets containing 50 mg
dinitrobenzene solution and 1 ml of sodium hydroxide of Mebendazole with 10 ml of a mixture of 10 volumes of
solution; an intense yellow colour is produced. anhydrous formic acid and 90 volumes of chloroform for
30 minutes, filter, evaporate the filtrate to dryness and dry the
D. Melts at about 290° (2.4.21).
residue at a pressure not exceeding 0.7 kPa.
Tests On the residue, determine by infrared absorption
Related substances. Determine by thin-layer chromatography obtained with mebendazole RS or with the reference spectrum
(2.4.17), coating the plate with silica gel GF254. of mebendazole.
Mobile phase. A mixture of 90 volumes of chloroform, 5 B. In the test for Related substances, the principal spot in the
volumes of methanol and chromatogram obtained with the test solution corresponds to
5 volumes of anhydrous formic acid. that in the chromatogram obtained with reference solution (a).
examination in 1 ml of anhydrous formic acid and add Tests
sufficient chloroform to produce 10 ml. Related substances. Determine by thin-layer chromatography
Reference solution (a). A 0.5 per cent w/v solution of (2.4.17), coating the plate with silica gel GF254.
mebendazole RS in a mixture of 10 volumes of anhydrous Mobile phase. A mixture of 90 volumes of chloroform,
formic acid and 90 volumes of chloroform. 5 volumes of methanol and 5 volumes of anhydrous formic
Reference solution (b). A 0.0025 per cent w/v solution of acid.
mebendazole RS in a mixture of 10 volumes of anhydrous Test solution. Shake a quantity of the powdered tablets
formic acid and 90 volumes of chloroform. containing 50 mg of Mebendazole with 10 ml of a mixture of
Apply to the plate 10 µl of each solution. After development, 10 volumes of anhydrous formic acid and 90 volumes of
dry the plate in air and examine in ultraviolet light at 254 nm. chloroform and filter.
721
MEBEVERINE HYDROCHLORIDE IP 2007
Reference solution (a). A 0.5 per cent w/v solution of hydrochloride RS or with the reference spectrum of
mebendazole RS in a mixture of 10 volumes of anhydrous mebeverine hydrochloride.
formic acid and 90 volumes of chloroform. B. When examined in the range 230 nm to 360 nm (2.4.7), a
Reference solution (b). A 0.0025 per cent w/v solution of 0.003 per cent w/v solution in 0.1 M hydrochloric acid shows
mebendazole RS in a mixture of 10 volumes of anhydrous an absorption maximum at about 263 nm and a less well-defined
formic acid and 90 volumes of chloroform. maximum at about 292 nm; absorbance at about 263 nm, about
0.79 and at about 292 nm, about 0.41.
dry the plate in air and examine in ultraviolet light at 254 nm. C. Dissolve 25 mg in 2 ml of water, acidify with 2 M nitric acid
Any secondary spot in the chromatogram obtained with the and centrifuge. The supernatant liquid gives the reactions of
test solution is not more intense than the spot in the chlorides (2.3.1).
Tests
Assay. Weigh and powder 20 tablets. Weigh accurately a pH (2.4.24). 4.5 to 6.5, determined in a 2.0 per cent w/v solution.
quantity of the powder containing about 50 mg of Ether-soluble extractive. Dissolve 40 mg in 25 ml of 2 M
Mebendazole, add 50 ml of 0.5 M methanolic hydrochloric hydrochloric acid and shake with 50 ml of ether for 1 minute.
acid shake for 30 minutes and dilute to 100.0 ml with 0.5 M Wash the ether layer with three quantities, each of 25 ml, of
methanolic hydrochloric acid. Filter and discard the first 10 water, evaporate the ether to dryness using a rotary evaporator
ml of the filtrate. Dilute 10.0 ml of the filtrate to 100.0 ml with and dissolve the residue in sufficient methanol to produce 20
0.5 M methanolic hydrochloric acid and mix. Further dilute ml; absorbance of the resulting solution at about 260 nm, not
5.0 ml to 50.0 ml with the same solvent and mix. Measure the more than 0.23 (2.4.7).
234 nm (2.4.7). Calculate the content of C16H13N3O3 from the Non-tertiary amine. Dissolve 0.5 g in 5 ml of pyridine, add 5
absorbance obtained by repeating the operation using ml of copper chloride-pyridine reagent and heat at 50° for 30
mebendazole RS in place of the substance under examination. minutes. Cool, add sufficient acetone to produce 50 ml and
measure the absorbance of the resulting solution at about 405
Storage. Store protected from light and moisture. nm (2.4.7), using as the blank a solution obtained by treating
5 ml of pyridine in the same manner. The absorbance is not
more than that obtained by repeating the test using 5 ml of a
Mebeverine Hydrochloride 0.006 per cent w/v solution of di-n-butylamine in pyridine
and beginning at the words “add 5 ml of copper chloride-
pyridine reagent....”.
CH3
O O OCH3
N , HCl (2.4.17), coating the plate with silica gel F254.
Mobile phase. A mixture of 50 volumes of ethanol, 50 volumes
H 3C
of chloroform and 1 volume of 18 M ammonia.
H3CO Test solution. Dissolve 0.2 g of the substance under
OCH3 examination in 10 ml of acetone.
C25H35NO5,HCl Mol. Wt. 466.0 Reference solution (a). Dissolve 10 mg of the substance under
Mebeverine Hydrochloride is (RS)-4-[ethyl(4-methoxy-α-
methylphenethyl)amino]butyl veratrate hydrochloride. Reference solution (b). A 0.002 per cent w/v solution of
veratric acid in acetone.
Mebeverine Hydrochloride contains not less than 99.0 per
cent and not more than 101.0 per cent of C25H35NO5,HCl, Apply to the plate 10 µl of each solution. After development,
calculated on the dried basis. dry the plate in air and examine in ultraviolet light at 254 nm.
Expose the plate to iodine vapour for 1 hour. When viewed in
Description. A white or almost white, crystalline powder. ultraviolet light, any spot corresponding to veratric acid in
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6). reference solution (b).Using both methods of visualisation
Compare the spectrum with that obtained with mebeverine any other secondary spot in the chromatogram obtained with
722
IP 2007 MECLIZINE HYDROCHLORIDE
the test solution is not more intense than the spot in the Reference solution (a). Dilute 1 volume of the test solution to
chromatogram obtained with reference solution (a). 200 volumes with acetone.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Reference solution (b). A 0.01 per cent w/v solution of veratric
acid in acetone.
on 1.0 g by drying in an oven at 105° for 1 hour. Apply to the plate 10 µl of each solution. After development,
Expose the plate to iodine vapour for 1 hour. When viewed in
ultraviolet light, any spot corresponding to veratric acid in
acetate solution. Titrate with 0.1 M perchloric acid,
reference solution (b). Using both methods of visualisation
1 ml of 0.1 M perchloric acid is equivalent to 0.04660 g of any other secondary spot in the chromatogram obtained with
C25H35NO5,HCl. the test solution is not more intense than the spot in the
Storage. Store protected from light and moisture at a chromatogram obtained with reference solution (a).
temperature not exceeding 30°. Other tests. Comply with the tests stated under Tablets.
quantity of the powder containing about 0.5 g of Mebeverine
Mebeverine Tablets Hydrochloride, dissolve in 100 ml of 0.1 M hydrochloric acid
and heat for 10 minutes on a water-bath, shaking occasionally.
Mebeverine Hydrochloride Tablets
Cool, add sufficient 0.1 M hydrochloric acid to produce
Mebeverine Tablets contain not less than 95.0 per cent and 250.0 ml and filter. To 10.0 ml of filtrate add sufficient 0.1 M
not more than 105.0 per cent of the stated amount of hydrochloric acid to produce 100.0 ml and dilute 10.0 ml of
mebeverine hydrochloride, C25H35NO5,HCl. The tablets are this solution to 100.0 ml with the same solvent. Measure the
coated. absorbance of the resulting solution at the maximum at about
263 nm (2.4.7). Calculate the content of C25H35NO5,HCl. taking
Identification 263 as the specific absorbance at 263 nm.
A. Suspend a quantity of the powdered tablets containing Storage. Store protected from light and moisture.
0.2 g of Mebeverine Hydrochloride in 20 ml of water, add 5 ml
of 5 M sodium hydroxide and extract with two quantities,
each of 25 ml, of chloroform. Dry the combined extracts over
anhydrous sodium sulphate and evaporate to dryness using Meclizine Hydrochloride
a rotary evaporator.
Meclozine Hydrochloride
On the residue, determine by infrared absorption
obtained with mebeverine hydrochloride RS or with the
reference spectrum of mebeverine hydrochloride.
CH3
solution obtained in the Assay shows an absorption maximum N , 2HCl
at about 263 nm and a less well-defined maximum at about N
292 nm. Cl
Tests C25H27ClN2, 2HCl Mol. Wt. 463.9

Related substances. Determine by thin-layer chromatography Meclizine Hydrochloride is (RS)-1-(4-chlorobenzhydryl)-4-
(2.4.17), coating the plate with silica gel F254. (3-methylbenzyl)piperazine dihydrochloride.
Mobile phase. A mixture of 50 volumes of ethanol, 50 volumes Meclizine Hydrochloride contains not less than 98.0 per cent
of chloroform and 1 volume of 18 M ammonia. and not more than 102.0 per cent of C25H27ClN2, 2HCl,
containing 0.2 g of Mebeverine Hydrochloride with 10 ml of Description. A white or yellowish white, crystalline powder;
acetone and filter. odour, slight.
723
MECLIZINE TABLETS IP 2007
Test A may be omitted if tests B, C and D are carried out. Tests Water (2.3.43). Not more than 5.0 per cent, determined on 0.2
B and C may be omitted if tests A and D are carried out. g.
A. Determine by infrared absorption spectrophotometry (2.4.6). Assay. Weigh accurately about 0.35 g, dissolve in 50 ml of
Compare the spectrum with that obtained with meclizine chloroform, add 50 ml of anhydrous glacial acetic acid, 5 ml
hydrochloride RS or with the reference spectrum of meclizine of acetic anhydride and 12 ml of mercuric acetate solution.
hydrochloride. Titrate with 0.1 M perchloric acid, using a 0.1 per cent w/v
solution of quinaldine red in anhydrous glacial acetic acid
B. When examined in the range 220 nm to 360 nm (2.4.7), a as indicator. Carry out a blank titration.
an absorption maximum at about 232 nm and weak absorption 1 ml of 0.1 M perchloric acid is equivalent to 0.02319 g of
without a defined maximum in the range 260 nm to 300 nm; C25H27ClN2, 2HCl.
absorbance at the maximum at about 232 nm, 0.51 to 0.57. Storage. Store protected from light and moisture.
chromatogram obtained with the test solution (b) corresponds
to that in the chromatogram obtained with reference solution Meclizine Tablets
(b).
Meclizine HydrochlorideTablets; Meclozine
D. Dissolve about 15 mg in 2 ml of ethanol (95 per cent); the
solution gives reaction A of chlorides (2.3.1). Hydrochloride Tablets; Meclozine Tablets
Meclizine Tablets contain not less than 95.0 per cent and not
Tests more than 105.0 per cent of the stated amount of meclizine
hydrochloride, C25H27ClN2, 2HCl.
(95 per cent) is clear (2.4.1), and not more intensely coloured Identification
than reference solution YS6 (2.4.1).
Triturate a quantity of the powdered tablets containing 0.5 g
of Meclizine Hydrochloride with three quantities, each of 15
ml, of chloroform. Filter the extracts and evaporate the clear
Mobile phase. A mixture of 60 volumes of dichloromethane, filtrate to dryness on a water-bath.The residue complies with
30 volumes of toluene, 5 volumes of methanol and 0.5 volume the following tests.
Test solution (a). Dissolve 0.5 g of the substance under Compare the spectrum with that obtained with meclizine
examination in 10 ml of a mixture of equal volumes of hydrochloride RS or with the reference spectrum of meclizine
dichloromethane and methanol. hydrochloride.
Test solution (b). Dissolve 0.5 g of the substance under B. When examined in the range 220 nm to 360 nm (2.4.7), a
examination in 100 ml of a mixture of equal volumes of 0.0015 per cent w/v solution in 0.1 M hydrochloric acid shows
dichloromethane and methanol. an absorption maximum at about 232 nm and weak absorption
Reference solution (a). Dissolve 25.0 mg of the substance without a defined maximum in the range 260 nm to 300 nm;
under examination in 100 ml of a mixture of equal volumes of absorbance at the maximum at about 232 nm, 0.51 to 0.57.
dichloromethane and methanol. C. Dissolve about 15 mg in 2 ml of ethanol (95 per cent); the
Reference solution (b). A 0.5 per cent w/v solution of solution gives reaction A of chlorides (2.3.1).
meclizine hydrochloride RS in a mixture of equal volumes of
Tests
Apply to the plate 10 µl of each solution. After development, Other tests. Comply with the tests stated under Tablets.
dry the plate in air and spray with dilute potassium Assay. Weigh and reduce to a fine powder 20 tablets. Weigh
iodobismuthate solution. Any secondary spot in the accurately a quantity of the powder containing about 0.35 g
chromatogram obtained with test solution (a) is not more of Meclizine Hydrochloride and extract with three quantities,
intense than the spot in the chromatogram obtained with each of 50 ml, of chloroform, stirring the mixture each time for
reference solution (a). Ignore any yellowish white spot on the 30 minutes, then allowing the undissolved matter to settle and
line of application. decanting the supernatant liquid on to a sintered-glass filter
724
IP 2007 MEDROXYPROGESTERONE ACETATE
(porosity No. 4). Transfer the residue to the filter with the aid Reference solution (a). A 0.1 per cent w/v solution of
of chloroform and wash the vessel and filter with 20 ml of medroxyprogesterone acetate RS in a mixture of 90 volumes
chloroform. Combine the extracts and washing and evaporate of chloroform and 10 volumes of methanol.
on a water-bath to 50 ml. Cool and add 50 ml of anhydrous
glacial acetic acid, 5 ml of acetic anhydride and 12 ml of
w/v each of progesterone RS and medroxyprogesterone
mercuric acetate solution. Titrate with 0.1 M perchloric acid,
acetate RS in a mixture of 90 volumes of chloroform and
using a 0.1 per cent w/v solution of quinaldine red in
anhydrous glacial acetic acid as indicator. Carry out a blank
titration. Apply to the plate 5 µl of each solution. After development,
1 ml of 0.1 M perchloric acid is equivalent to 0.02319 g of dry the plate in air and examine in ultraviolet light at 254 nm.
C25H27ClN2, 2HCl. The principal spot in the chromatogram obtained with the test
Storage. Store protected from light and moisture. with reference solution (a). The test is not valid unless the
chromatogram obtained with reference solution (b) shows two
clearly separated spots.
Medroxyprogesterone Acetate Spray with ethanolic sulphuric acid (20 per cent), heat at
120° for 10 minutes or until spots appear and allow to cool.
Examine the plate in ultraviolet light at 365 nm. The principal
O spot in the chromatogram obtained with the test solution
H3 C CH3
O CH3
H3 C H chromatogram obtained with reference solution (b) shows two
O
clearly separated spots.
H H
O C. Melting range 205° to 209° (2.4.21).
CH3
Tests
C24H34O4 Mol. wt. 386.5 Specific optical rotation (2.4.22). +45.0° to +51.0°, determined
Medroxyprogesterone Acetate is 6α-methyl-3,20- in a 1.0 per cent w/v solution in dioxan.
dioxopregn-4-en-17α-yl acetate. Related substances. Determine by liquid chromatography
Medroxyprogesterone Acetate contains not less than 97.0 (2.4.17).
per cent and not more than 103.0 per cent of C24H34O4, Test solution (a). Dissolve 5 mg of the substance under
calculated on the dried basis. examination in 100 ml of the mobile phase.
Description. A white or almost white, crystalline powder. Test solution (b). Dissolve 0.25 g of the substance under
Identification
Reference solution. Evaporate 1 ml of a solution containing
Test A may be omitted if tests B and C are carried. Test B may
0.1 per cent w/v each of medroxyprogesteronge acetate RS
be omitted if tests A and C are carried out.
and megestrol acetate RS in ethanol, to dryness in a water
A. Determine by infrared absorption spectrophotometry (2.4.6). bath at 45° and dissolve the residue in sufficient mobile phase
Compare the spectrum with that obtained with to produce 25 ml.
medroxyprogesterone acetate RS.
B. Determine by thin-layer chromatography (2.4.17), coating – a stainless steel column 25 cm x 4.6 mm, packed with
the plate with silica gel GF254. octadecylsilane bonded to porous silica (5 µm),
Mobile phase. A mixture of 70 volumes of toluene, 40 volumes – mobile phase: a mixture of 600 volumes of acetonitrile
of ethyl acetate and 10 volumes of light petroleum (50° to and 350 volumes of water and diluted to 1000 volumes
70°). with water,
examination in 100 ml of a mixture of 90 volumes of chloroform – spectrophotometer set at 254 nm,
and 10 volumes of methanol. – a 20 µl loop injector.
725
MEFENAMIC ACID IP 2007
Adjust the sensitivity so that the height of the principal peak Mefenamic Acid contains not less than 99.0 per cent and not
in the chromatogram obtained with test solution (a) is 70 to 90 more than 100.5 per cent of C15H15NO2, calculated on the dried
per cent of the full-scale deflection. basis.
Equilibrate the column with the mobile phase at a flow rate of Description. A white to greyish-white, microcrystalline
1 ml per minute for about 45 minutes. When the chromatograms powder; odourless or almost odourless.
are recorded under the conditions described above, the
retention times are 12.5 minutes for megestrol acetate and Identification
13.5 minutes for medroxyprogesterone acetate.
Inject the reference solution. The test is not valid unless the Compare the spectrum with that obtained with mefenamic acid
resolution between the peaks due to megestrol acetate and RS or with the reference spectrum of mefenamic acid.
medroxyprogesterone acetate is at least 2.0. If this resolution
B. Dissolve 25 mg in 15 ml of chloroform and examine in
is not achieved, adjust the concentration of acetonitrile in
ultraviolet light at 365 nm; the solution exhibits a strong
the mobile phase. Verify the repeatability by making five
greenish-yellow fluorescence. Carefully add 0.5 ml of a
separate injections of test solution (a). The system is not
saturated solution of trichloroacetic acid drop wise and
suitable unless the relative standard deviation for the area of
examine again in ultraviolet light at 365 nm; the solution does
the principal peak in the chromatogram obtained with test
not exhibit fluorescence.
solution (a). The system is not suitable unless the relative
standard deviation for the area of the principal peak in the C. Dissolve 5 mg in 2 ml of sulphuric acid and add 0.05 ml of
chromatogram obtained with test solution (a) is less than 0.0167 M potassium dichromate; an intense blue colour is
2.0 per cent. produced immediately which fades rapidly to brownish-green.
Inject separately test solutions (a) and (b) and record the Tests
chromatograms for 1.5 times the retention time of the principal
peak. In the chromatogram obtained with test solution (b) the Light absorption (2.4.7). Absorbance of a 0.002 per cent w/v
area of any secondary peak is not greater than half the area of solution in a mixture of 99 volumes of methanol and 1 volume
the principal peak in the chromatogram obtained with test of 1 M hydrochloric acid at the maximum at about 279 nm,
solution (a) and the sum of the areas of the secondary peaks 0.69 to 0.74 and at the maximum at about 350 nm, 0.56 to 0.60.
is not greater than the area of the principal peak in the Copper. Moisten 1.0 g with sulphuric acid and ignite until all
chromatogram obtained with test solution (a). Ignore any peak the carbon is removed. Add 10 ml of 1 M sulphuric acid to the
the area of which is less than 0.025 times the area of the principal residue and allow to stand for 10 minutes. Transfer to a
peak in the chromatogram obtained with test solution (a). separating funnel using 20 ml of water and add 10 ml of a
Loss on drying (2.4.19). Not more than 1.0 per cent, determined solution containing 20 per cent w/v diammonium hydrogen
on 0.5 g by drying in an oven at 105° for 3 hours. citrate and 5 per cent w/v solution of disodium edetate. Add
0.2 ml of thymol blue solution and neutralise with 5 M
Assay. Weigh accurately about 25 mg, dissolve in sufficient ammonia. Add 10 ml of sodium diethyldithiocarbamate
ethanol (95 per cent) and measure the absorbance of the solution and 15 ml of carbon tetrachloride, shake and allow
resulting solution at the maximum at about 241 nm (2.4.7). to separate. The yellow colour of the carbon tetrachloride
Calculate the content of C24H34O4 taking 426 as the specific layer is not more intense than that produced by treating 2 ml
absorbance at 241 nm. of copper standard solution (10 ppm Cu) in the same manner
Storage. Store protected from light and moisture. beginning at the words “Transfer to a separating funnel
using.....” (20 ppm).
Mefenamic Acid (2.4.17), coating the plate with silica gel GF254.
HOOC CH3 of dioxan and 1 volume of glacial acetic acid.
H
N CH3 Test solution. Dissolve 0.25 g of the substance under
examination in 10 ml of a mixture of 3 volumes of chloroform
Reference solution. Dissolve 5.0 mg of the substance under
C15H15NO2 Mol. Wt. 241.3
Mefenamic Acid is N-(2,3-xylyl)anthranilic acid. and 1 volume of methanol.
726
IP 2007 MEFENAMIC ACID CAPSULES
Apply to the plate 20 µl of each solution. After development, Identification

dry the plate in air, expose to iodine vapour for 5 minutes and
examine in ultraviolet light at 254 nm. Any secondary spot in Extract a quantity of the contents of the capsules containing
the chromatogram obtained with the test solution is not more 0.25 g of Mefenamic Acid with two quantities, each of 30 ml, of
intense than the spot in the chromatogram obtained with the ether. Wash the combined extracts with water and evaporate
reference solution. to dryness on a water-bath. The residue, after drying at 105°,
complies with the following tests.
2,3-Dimethylaniline. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel G. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with mefenamic acid
RS or with the reference spectrum of mefenamic acid.
of dioxan and 1 volume of 18 M ammonia.
B. Dissolve 25 mg in 15 ml of chloroform and examine in
ultraviolet light at 365 nm; the solution exhibits a strong
greenish-yellow fluorescence. Carefully add 0.5 ml of a
saturated solution of trichloroacetic acid drop wise and
Reference solution. A 0.00025 per cent w/v solution of 2,3- examine again in ultraviolet light at 365 nm; the solution does
dimethylaniline in a mixture of 3 volumes of chloroform and not exhibit fluorescence.
1 volume of methanol.
C. Dissolve 5 mg in 2 ml of sulphuric acid and add 0.05 ml of
Apply to the plate 40 µl of each solution. After development, 0.0167 M potassium dichromate; an intense blue colour is
dry the plate in a current of warm air. Spray the plate with produced immediately which fades rapidly to brownish-green.
ethanolic sulphuric acid (20 per cent), heat at 105° for
30 minutes and immediately expose to nitrous fumes in a closed Tests
glass chamber for 15 minutes (the nitrous fumes may be
generated by adding dilute sulphuric acid dropwise to a Related substances. Determine by thin-layer chromatography
solution containing 10 per cent w/v of sodium nitrite and (2.4.17), coating the plate with silica gel GF254.
3 per cent w/v of potassium iodide). Place the plate in a current
of warm air for 15 minutes and spray with a 0.5 per cent w/v
of dioxan and 1 volume of glacial acetic acid.
solution of N-(1-naphthyl)ethylenediamine dihydrochloride
in ethanol (95 per cent). If necessary, allow to dry and repeat Test solution. The supernatant liquid obtained by shaking a
the spraying. Any spot corresponding to 2,3-dimethylaniline quantity of the contents of the capsules containing 0.25 g of
in the chromatogram obtained with the test solution is not Mefenamic Acid with a mixture of 7.5 ml of chloroform and
more intense than the spot in the chromatogram obtained 2.5 ml of methanol.
Reference solution. Dissolve 5.0 mg of the substance under
Sulphated ash (2.3.18). Not more than 0.1 per cent. examination in 100 ml of a mixture of 3 volumes of chloroform
Loss on drying (2.4.19). Not more than 1.0 per cent, determined and 1 volume of methanol.
on 1.0 g by drying in an oven at 105°. Apply to the plate 20 µl of each solution. After development,
Assay. Weigh accurately about 0.5 g and dissolve in 100 ml of dry the plate in air, expose to iodine vapour for 5 minutes and
warm ethanol previously neutralised to phenol red solution examine in ultraviolet light at 254 nm. Any secondary spot in
and titrate with 0.1 M sodium hydroxide using phenol red the chromatogram obtained with the test solution is not more
solution as indicator. intense than the spot in the chromatogram obtained with the
reference solution.
C15H15NO2. 2,3-Dimethylaniline. Determine by thin-layer chromatography
of dioxan and 1 volume of 18 M ammonia.
Test solution. The supernatant liquid obtained in the test for
Mefenamic Acid Capsules Related substances.
Mefenamic Acid Capsules contain not less than 90.0 per cent Reference solution. A 0.00025 per cent w/v solution of 2,3-
and not more than 110.0 per cent of the stated amount of dimethylaniline in a mixture of 3 volumes of chloroform and 1
mefenamic acid, C15H15NO2. volume of methanol.
727
MEGESTROL ACETATE IP 2007
Apply to the plate 40 µl of each solution. After development, – mobile phase: a mixture of 23 volumes acetonitrile,
dry the plate in a current of warm air. Spray the plate with 20 volumes of 0.05 M of monobasic ammonium
ethanolic sulphuric acid (20 per cent), heat at 105° for 30 phosphate adjusted to a pH of 5.0 with 3 M ammonia
minutes and immediately expose to nitrous fumes in a closed and 7 volumes of tetrahydrofuran,
glass chamber for 15 minutes (the nitrous fumes may be – flow rate. 1 ml per minute,
generated by adding dilute sulphuric acid dropwise to a – spectrophotometer set at 254 nm,
solution containing 10 per cent w/v of sodium nitrite and 3 – a 20 µl loop injector.
per cent w/v of potassium iodide). Place the plate in a current Inject the reference solution. The test is not valid unless the
of warm air for 15 minutes and spray with a 0.5 per cent w/v column efficiency determined from the mefenamic acid peak is
solution of N-(1-naphthyl)ethylenediamine dihydrochloride not less than 8200 theoretical plates, the tailing factor is not
in ethanol (95 per cent). If necessary, allow to dry and repeat more than 1.6 and the relative standard deviation for replicate
the spraying. Any spot corresponding to 2,3-dimethylaniline injections is not more than 1.0 per cent.
more intense than the spot in the chromatogram obtained Inject alternately the test solution and the reference solution.
with the reference solution. Calculate the content of C15H5NO2 in the capsules.
Apparatus. No 2
Medium. 900 ml of 0.05 M tris buffer prepared by dissolving
60.5 g tris(hydroxymethyl) aminomethane in 6000 ml water,
diluting to 10,000 ml with water and adjusting with phosphoric
Megestrol Acetate
acid to a pH 9.0 ± 0.05. 100 g of sodium lauryl sulphate is
dissolved in 6000 ml of the above solution and further mixed O
H3C CH3
with the remaining quantity of the solution.
O CH3
H3C H
Withdraw a suitable volume of the medium and filter. O
Determine by liquid chromatography (2.4.14). H H
O
Test solution. Use the filtrate as the test solution.
CH3
Reference solution. A 0.02 per cent w/v solution of mefenamic
acid RS in the dissolution medium. C24H32O 384.5
Use the chromatographic system described under Assay. Megestrol Acetate is 6-methyl-3,20-dioxopregna-4,6-dien-
Calculate the content of C15H15NO2 in the medium. 17-yl acetate.
D. Not less than 75 per cent of the stated amount of C15H15NO2. Megestrol Acetate contains not less than 97.0 per cent and
Other tests. Comply with the tests stated under Capsules. dried basis.
Assay. Determine by liquid chromatography (2.4.14). Description. A white to creamy-white, crystalline powder;
Test solution. Weigh accurately a quantity of the mixed odourless or almost odourless.
contents of 20 capsules containing about 50 mg of Mefenamic
acid in a 250.0 ml volumetric flask, add 5 ml of tetrahydrofuran, Identification
shake for 10 minutes with the aid of ultrasound, dilute to A. Determine by infrared absorption spectrophotometry (2.4.6).
volume with the mobile phase and filter. Compare the spectrum with that obtained with megestrol
Reference solution. A solution containing 0.02 per cent w/v of acetate RS or with the reference spectrum of megestrol acetate.
mefenamic acid RS in the mobile phase. B. Determine by thin-layer chromatography (2.4.17), coating
Protect the solutions from light. the plate with silica gel G.
Chromatographic system Solvent mixture. A mixture of 90 volumes of acetone and 1,2-

– a stainless steel column 25 cm x 4.6 mm, packed with propanediol.
octadecylsilane chemically bonded to porous silica Mobile phase. A mixture of 40 volumes of cyclohexane and
(5 µm), 10 volumes of toluene.
728
IP 2007 MEGESTROL TABLETS
Test solution. Dissolve 25 mg of the substance under intense than the spot in the chromatogram obtained with the
examination in 10 ml of the solvent mixture. reference solution.
Reference solution (a). Dissolve 25 mg of megestrol acetate Heavy metals (2.3.13). 1.0 g complies with limit test for heavy
RS in 10 ml of the solvent mixture. metals, Method B (20 ppm).
Reference solution (b). Mix equal volumes of the test solution Sulphated ash (2.3.18). Not more than 0.1 per cent.
Place the dry plate in a tank containing a shallow layer of the on 1.0 g by drying in an oven at 105°.
solvent mixture, allow the solvent mixture to ascend to the Assay. Weigh accurately about 20 mg, dissolve in sufficient
top, remove the plate from the tank and allow the solvent to ethanol to produce 100.0 ml, dilute 5.0 ml to 100.0 ml with
evaporate. Use within 2 hours, with the flow of the mobile ethanol and measure the absorbance of the resulting solution
phase in the direction in which the aforementioned treatment at the maximum at about 287 nm (2.4.7). Calculate the content
was done. of C24H32O4 taking 630 as the specific absorbance at 287 nm.
Heat at 120° for a further 10 minutes, allow to cool and examine Megestrol Tablets
spot in the chromatogram obtained with the test solution Megestrol AcetateTablets
corresponds to that in the chromatogram obtained with Megestrol Tablets contain not less than 92.5 per cent and not
reference solution (a). The principal spot in the chromatogram more than 107.5 per cent of megestrol acetate, C24H32O4.
compact spot. Identification
C. Gives the reaction of acetyl groups (2.3.1). Extract a quantity of the powdered tablets containing 40 mg of
Megestrol Acetate with 10 ml of chloroform, filter and
Tests evaporate the filtrate to dryness in a current of air. Dry the
Specific optical rotation (2.4.22). +9.0° to +12.0°, determined residue at 60° at a pressure not exceeding 0.7 kPa for 1 hour.
at 20° in a 5.0 per cent w/v solution in chloroform. On the residue, determine by infrared absorption
Light absorption. When examined in the range 230 nm to spectrophotometry (2.4.6). Compare the spectrum with that
360 nm (2.4.7), the solution obtained in the Assay shows an obtained with megestrol acetate RS or with the reference
absorption maximum only at about 287 nm; ratio of the spectrum of megestrol acetate.
absorbance at about 240 nm to that at the maximum at about
287 nm, not more than 0.17.
Tests
Related foreign steroids. Determine by thin-layer Disintegration (2.5.1). 30 minutes.

chromatography (2.4.17), coating the plate with silica gel G. Other tests. Comply with the tests stated under Tablets.
Mobile phase. A mixture of 92 volumes of 1,2-dichloroethane, Assay. Weigh and powder 20 tablets. Weigh accurately a
8 volumes of methanol and 0.5 volume of water. quantity of the powder containing about 50 mg of Megestrol
Test solution. Dissolve 0.5 g of the substance under Acetate and dissolve as completely as possible in sufficient
examination in 10 ml of a mixture of 9 volumes of chloroform methanol to produce 100.0 ml. Mix well and filter. Dilute 2.0 ml
and 1 volume of methanol. of the filtrate with methanol to 100.0 ml and mix. Measure the
Reference solution. A 0.025 per cent w/v solution of megestrol 287 nm (2.4.7). Calculate the content of C24H32O4 taking 630 as
RS in a mixture of 9 volumes of chloroform and 1 volume of the specific absorbance at 287 nm.
methanol.
sulphuric acid in ethanol, heat at 110° for 10 minutes and
729
MELPHALAN IP 2007
Melphalan silver nitrate, determining the end-point potentiometrically

(2.4.25). Subtract the equivalent volume of 0.1 M silver nitrate
used in the test for Ionisable chlorine. The difference between
O
the volumes represents the amount of 0.1 M silver nitrate
OH required by melphalan.
Cl NH2 1 ml of 0.1 M silver nitrate is equivalent to 0.01526 g of
N C13H18Cl2N2O2.
Cl
C13H18Cl2N2O2 Mol. Wt. 305.2

Melphalan is 4-bis(2-chloroethyl)amino-L-phenylalanine. Melphalan Injection
Melphalan contains not less than 93.0 per cent and not more
Melphalan Injection is a sterile material consisting of
than 100.5 per cent of C13H18Cl2N2O2, calculated on the dried
Melphalan Hydrochloride with or without auxiliary
basis.
substances. It is filled in a sealed container.
Description. A white or almost white powder; odourless or
almost odourless.
Identification Injections, immediately before use.
The constituted solution complies with the requirement for
A. When examined in the range 230 nm to 360 nm (2.4.7), a
Particulate matter stated under Parenteral Preparations
(Injections) and with the following tests.
maximum at about 260 nm and a less well-defined maximum at
about 301 nm. Appearance of solution. Not more opalescent than opalescence
standard OS2 (2.4.1).
B. Dissolve 20 mg in 50 ml of methanol with the aid of gentle
heat, add 1 ml of a 5 per cent w/v solution of 4-(4- pH (2.4.24). 6.0 to 7.0.
nitrobenzyl)pyridine in acetone and evaporate to dryness. Storage. The constituted solution should be used immediately
Dissolve the residue in 1 ml of hot methanol and add 0.1 ml of after preparation but, in any case, within the period
strong ammonia solution; a red colour is produced. recommended by the manufacturer.
C. Heat 0.1 g with 10 ml of 0.1 M sodium hydroxide for 10 Melphelan Injection contains not less than 90.0 per cent and
minutes on a water-bath. The resulting solution, after not more than 110.0 per cent of the stated amount of
acidification with 2 M nitric acid, gives reaction A of chlorides anhydrous melphalan, C13H18Cl2N2O2.
(2.3.1).
Tests requirements stated under Parenteral Preparations
in a 0.7 per cent w/v solution in methanol prepared with the Identification
aid of gentle heat.
A. When examined in the range 230 nm to 360 nm (2.4.7), a
Ionisable chlorine. Dissolve 0.4 g in a mixture of 75 ml of 0.001 per cent w/v solution in methanol shows an absorption
water and 2 ml of nitric acid. Allow to stand for 2 minutes and maximum at about 260 nm and a less well-defined maximum at
titrate with 0.1 M silver nitrate, determining the end-point about 301 nm.
potentiometrically (2.4.25); not more than 0.8 ml is required.
Sulphated Ash (2.3.18). Not more than 0.3 per cent. obtained with the test solution corresponds to the peak in the
Loss on drying (2.4.19). Not more than 7.0 per cent, determined chromatogram obtained with the reference solution.
on 0.5 g by drying in an oven at 105° at a pressure not exceeding C. Dissolve a quantity containing 20 mg of anhydrous
0.7 kPa for 2 hours. melphalan in 50 ml of methanol with the aid of gentle heat, add
Assay. Weigh accurately about 0.4 g, add 20 ml of a 20 per cent 1 ml of a 5 per cent w/v solution of 4-(4-nitrobenzyl)pyridine
w/v solution of potassium hydroxide, heat on a water-bath in acetone and evaporate to dryness. Dissolve the residue in
for 2 hours, replacing the water lost by evaporation. Cool, add 1 ml of hot methanol and add 0.1 ml of strong ammonia
75 ml of water and 4 ml of nitric acid, cool. Titrate with 0.1 M solution; a red colour is produced.
730
IP 2007 MELPHALAN TABLETS
D. Heat a quantity of the powder containing 0.1 g of anhydrous When examined in the range 230 nm to 360 nm (2.4.7), the
melphalan with 10 ml of 0.1 M sodium hydroxide for 10 minutes filtrate shows an absorption maximum at about 260 nm and a
on a water-bath. The resulting solution, after acidification with less well-defined maximum at about 301 nm.
2 M nitric acid, gives reaction A of chlorides (2.3.1).
B. To the remainder of the filtrate obtained in test A add 1 ml of
Tests a 5 per cent w/v solution of 4-(4-nitrobenzyl)pyridine in
acetone and evaporate to dryness. Dissolve the residue in
Ionisable chlorine. Dissolve a quantity containing 0.1 g of 1 ml of hot methanol and 0.1 ml of strong ammonia solution;
anhydrous melphalan in a mixture of 15 ml of water and 0.5 ml a red colour is produced.
of nitric acid, allow to stand for 2 minutes and titrate with
0.02 M silver nitrate determining the end-point Tests
potentiometrically (2.4.25); not more than 1.7 ml is required.
Tablets.
Test solution. Weigh accurately a quantity of the mixed
contents of 10 containers containing about 50 mg of Determine by liquid chromatography (2.4.14)
Melphalan, dissolve in a mixture of 4 volumes of acetonitrile Test solution. Add 20 ml of a mixture of 4 volumes of
and 1 volume of 0.1 M hydrochloric acid and dilute with acetonitrile and 1 volume of 0.1 M hydrochloric acid to one
sufficient of the same solvent mixture to produce a final solution tablet, mix with the aid of ultrasound for 10 minutes or until the
containing the equivalent of 0.01 per cent w/v of anhydrous tablet disintegrates, filter, discarding the first 5 ml of filtrate,
melphalan. and use the filtrate.
Reference solution. A 0.01 per cent w/v solution of melphalan Reference solution. A 0.01 per cent w/v solution of melphalan
RS in the same solvent. RS in the same solvent.
µm),
– mobile phase: a mixture of 200 volumes of a 0.375 per
– mobile phase: 200 volumes of a 0.375 per cent w/v
cent w/v solution of ammonium carbonate, 180 volumes
solution of ammonium carbonate, 180 volumes of
of methanol and 2.7 volumes of glacial acetic acid,
methanol and 2.7 volumes of glacial acetic acid,
Calculate the amount of C13H18Cl2N2O2 in the injection. Calculate the content of C13H18Cl2N2O2 in the tablet.
Storage. Store protected from light and moisture at a Other tests. Comply with the tests stated under Tablets.
temperature not exceeding 30°. Assay. Determine by liquid chromatography (2.4.14).
Labelling. The label on the sealed container states (1) the
Test solution. Weigh and powder 20 tablets and add about
equivalent amount of anhydrous melphalan contained in it;
150 ml of a mixture of 4 volumes of acetonitrile and 1 volume
(2) that it should be used immediately after preparation.
of 0.1 M hydrochloric acid to an accurately weighed quantity
of the powdered tablets containing about 25 mg of Melphalan,
shake and mix with the aid of ultrasound for 5 minutes, dilute
Melphalan Tablets to 250 ml with the same solvent. Filter, discarding the first
Melphalan Tablets contain not less than 90.0 per cent and not 20 ml of filtrate, and use the filtrate.
more than 110.0 per cent of the stated amount of melphalan, Reference solution. A 0.01 per cent w/v solution of melphalan
C13H18Cl2N2O2. The tablets are coated. RS in the same solvent.
Identification Use the chromatographic system described under Uniformity
of content.
A. Shake a quantity of the powdered tablets containing about
5 mg of Melphalan with 100 ml of hot methanol. Filter and Calculate the content of C13H18Cl2N2O2 in the tablets (for tablets
dilute 10 ml of the filtrate to 50 ml with methanol. containing more than 2 mg of Melphalan).
731
MENADIONE IP 2007
For tablets containing 2 mg or less of Melphalan, use the Reference solution. Dissolve 10 mg of the substance under
average of 10 individual results obtained in the test for examination in 100 ml of acetone.
Uniformity of content. Apply to the plate 5 µl of each solution. After development,
Storage. Store protected from light and moisture in a cool dry the plate in a current of hot air and repeat the development
place. and drying twice. Examine the plate in ultraviolet light at
254 nm. Any secondary spot in the chromatogram obtained
with the test solution is not more intense than the spot in the
Menadione
Menaphthone; Vitamin K3
O of 2 to 3 kPa for 4 hours.
CH3 Assay. Weigh accurately about 0.15 g, dissolve in 15 ml of
glacial acetic acid in a flask with a stopper fitted with a valve,
add 15 ml of 2 M hydrochloric acid and 1 g of zinc powder,
O close the flask and allow to stand in the dark for 60 minutes,
shaking occasionally. Filter the solution through absorbent
C11H8O2 Mol. Wt. 172.2 cotton and wash the filter with three quantities, each of 10 ml,
of carbon dioxide-free water, adding the washings to the
Menadione is 2-methyl-1,4-naphthaquinone.
filtrate. Add 0.1 ml of ferroin solution and immediately titrate
Menadione contains not less than 98.5 per cent and not more the combined filtrate and washings with 0.1 M ceric
than 101.0 per cent of C11H8O2, calculated on the dried basis. ammonium sulphate.
Description. A pale yellow, crystalline powder; odour, faint 1 ml of 0.1 M ceric ammonium sulphate is equivalent to
and characteristic. It decomposes on exposure to light, 0.00861 g of C11H8O2.
darkening in colour to light brown.
Identification
Menthol
A. Determine by infrared absorption spectrophotometry (2.4.6). CH3
Compare the spectrum with that obtained with menadione RS
or with the reference spectrum of menadione.
B. When examined in the range 230 nm to 360 nm (2.4.7), a OH
0.0005 per cent w/v solution in ethanol (95 per cent) shows
an absorption maximum only at about 250 nm. H 3C CH3
C. Dissolve about 1 mg in 5 ml of ethanol (95 per cent) and C10H20O Mol. Wt.156.3
add 2 ml of dilute ammonia solution and 0.2 ml of ethyl
cyanoacetate; an intense bluish violet colour develops which Menthol is 2-isopropyl-5-methylcyclohexanol. It is obtained
disappears on the addition of 2 ml of hydrochloric acid. from the volatile oils of various species of Mentha or
prepared synthetically. It may be laevo-rotatory [(–)-
Tests menthol] or racemic [(± )-menthol].
Related substances. Determine by thin-layer chromatography Description. Colourless, hexagonal or needle-like crystals, or
(2.4.17), coating the plate with silica gel GF254. infused masses or a crystalline powder; odour, pleasant and
characteristic.
5 volumes of 1,2-dichloroethane, 2 volumes of acetone and Identification
1 volume of nitromethane.
Test solution. Dissolve 0.2 g of the substance under Compare the spectrum with that obtained with menthol RS or
examination in 10 ml of acetone. with the reference spectrum of menthol.
732
IP 2007 MEPHENTERMINE SULPHATE
B. Dissolve 10 mg in 1 ml of sulphuric acid and add 1 ml of a the principal peak in the chromatogram obtained with reference
1 per cent w/v solution of vanillin in sulphuric acid; an solution (b).
orange-yellow colour is produced. Add 1 ml of water; the Residue on evaporation. Evaporate 2.0 g on a water-bath and
colour changes to violet (distinction from thymol). heat at 105° for 1 hour. The residue weighs not more than
C. When triturated with about an equal weight of camphor or 1.0 mg (0.05 per cent).
chloral hydrate or phenol, the mixture liquefies.
Tests temperature not exceeding 30°.
Labelling. The label states whether the contents are laevo-
Appearance of solution. Dissolve 1.0 g in 10 ml of ethanol rotatory or racemic menthol.
(95 per cent). The solution is not more opalescent than
opalescence standard OS4 (2.4.1), and not more intensely
coloured than reference solution RS6 (2.4.1).
Acidity. To 1.0 g in a 100-ml glass-stoppered conical flask add Mephentermine Sulphate
20 ml of water, boil until dissolution is complete, cool, stopper
the flask and shake vigorously for 1 minute. Add a few crystals
of the substance under examination to initiate crystallisation, H
shake vigorously for 1 minute and filter. To 5 ml of the filtrate N
CH3 , H2SO4, 2H2O
add 0.05 ml of methyl red solution and 0.05 ml of 0.01M sodium H3C CH3
hydroxide; the solution is yellow. 2
Specific optical rotation (2.4.22). (for (–)-menthol) –49.0° to –
51.0°; (for (± )-menthol) –2.0 ° to +2.0°, determined in a 10.0 (C11H17N)2,H2SO4,2H2O Mol. Wt.460.6
per cent w/v solution in ethanol (95 per cent).
Mephentermine Sulphate is N,α,α-trimethylphenethylamine
Congealing range (2.4.10). (for (± )-menthol) 27.0° to 28.0°; on sulphate dihydrate.
prolonged stirring, the temperature rises 30° to 32°.
Mephentermine Sulphate contains not less than 98.0 per cent
Related substances. Determine by gas chromatography and not more than 102.0 per cent of (C11H17N)2,H 2SO4,
(2.4.13). calculated on the dried basis.
Test solution. Dissolve 0.1 g in sufficient ethanol (95 per Description. White crystals or a crystalline powder.
cent) to produce 10 ml.
Identification
ml with ethanol (95 per cent). A. Determine by infrared absorption spectrophotometry (2.4.6).
Reference solution (b). Dilute 1 ml of reference solution (a) to Compare the spectrum with that obtained with mephentermine
20 ml with ethanol (95 per cent). sulphate RS.
Chromatographic system B. A 0.2 per cent w/v solution yields a precipitate with iodine
– a glass or stainless steel column 4 m x 2 mm, packed with solution and with potassium mercuri-iodide solution.
diatomaceous support (125 to 180 mesh) impregnated C. Dissolve 0.1 g in 5 ml of water, add with stirring 10 ml of
with a mixture suitable for the separation of free fatty picric acid solution. Allow to stand for 30 minutes, filter and
acids, wash the precipitate with small quantities of cold water until
– temperature : the last washing is colourless; the precipitate, after drying at
column 80°, 105° melts at 154° to 158° (2.4.21).
injection port at 250° and the detector at 240°,
Inject separately 1µl of each solution, and, after 2 minutes, Tests
increase the temperature of the column to 240° at a rate of 8°
per minute and maintain at this temperature for 15 minutes.
In the chromatogram obtained with the test solution the sum
of the areas of any secondary peaks is not greater than the
area of the principal peak in the chromatogram obtained with Loss on drying (2.4.19). 5.0 to 8.0 per cent, determined on
reference solution (a). Ignore any peak with an area less than 0.5 g by drying in an oven at 105° for 3 hours.
733
MEPHENTERMINE INJECTION IP 2007
Assay. Weigh accurately about 0.4 g, dissolve in 250 ml of 10 ml of ether, adding this ether to the main ether extract. Add
water, add 5 g of sodium chloride, shake well and add 5 ml of to the ether solution 30.0 ml of 0.05 M sulphuric acid, stir
5 M sodium hydroxide. Extract with 30 ml and then with further thoroughly and warm gently until the ether is evaporated.
quantities, each of 20 ml, of ether until the base is completely Cool and titrate with 0.1 M sodium hydroxide using methyl
extracted. Combine the ether extracts, wash with two quantities, red solution as indicator. Repeat the operation without the
each of 10 ml, of water and extract the aqueous washings with substance under examination. The difference between the
10 ml of ether, adding this ether to the main ether extract. Add titrations represents the amount of sulphuric acid required.
to the ether solution 30.0 ml of 0.05 M sulphuric acid, stir 1 ml of 0.05 M sulphuric acid is equivalent to 0.0163 g of
thoroughly and warm gently until the ether is evaporated. C11H17N.
Cool and titrate with 0.1 M sodium hydroxide using methyl
red solution as indicator. Repeat the operation without the Storage. Store protected from light and moisture.
substance under examination. The difference between the Labelling. The label states the strength in terms of the
titrations represents the amount of sulphuric acid required. equivalent amount of mephentermine in a suitable dose-volume.
1 ml of 0.05 M sulphuric acid is equivalent to 0.02123 g of
(C11H17N)2,H2SO4.
Storage. Store protected from light and moisture. Mepyramine Maleate
Pyrilamine Maleate
Mephentermine Injection
Mephentermine Sulphate Injection N N COOH
Mephentermine Injection is a sterile solution of ,
Mephentermine Sulphate in Water for Injections. H3CO
COOH
N
Mephentermine Injection contains not less than 95.0 per cent H 3C CH3
mephentermine, C11H17N. C17H23N3O,C4H4O4 Mol. Wt. 401.5
Mepyramine Maleate is 2-(N-4-anisyl-N-2-
Identification pyridylamino)ethyldimethylamine hydrogen maleate.
A. A 0.2 per cent w/v solution yields a precipitate with iodine Mepyramine Maleate contains not less than 99.0 per cent and
solution and with potassium mercuri-iodide solution. not more than 101.0 per cent of C17H23N3O,C4H4O4, calculated
B. Dissolve 0.1 g in 5 ml of water, add with stirring 10 ml of on the dried basis.
picric acid solution. Allow to stand for 30 minutes, filter and Description. A white or slightly yellowish, crystalline powder;
wash the precipitate with small quantities of cold water until odourless or almost odourless.
the last washing is colourless; the precipitate, after drying at
105° melts at 154° to 158° (2.4.21). Identification
C. Gives the reactions of sulphates (2.3.1). Test A may be omitted if tests B and C are carried out. Tests B
Tests
pH (2.4.24). 4.0 to 6.5. Compare the spectrum with that obtained with mepyramine
Other tests. Complies with the tests stated under Parenteral maleate RS or with the reference spectrum of mepyramine
Preparations (Injections). maleate.
Assay. Measure accurately a volume containing about 0.2 g B. When examined in the range 220 nm to 360 nm (2.4.7), a
of mephentermine, add water if necessary to produce 20 ml 0.001 per cent w/v solution in 0.01 M hydrochloric acid shows
add 5 g of sodium chloride, shake well and add 5 ml of 5 M absorption maxima, at about 239 nm and 316 nm; absorbance
sodium hydroxide. Extract with 30 ml and then with further at about 239 nm, 0.43 to 0.477 and at about 316 nm, 0.2 to 0.22.
quantities, each of 20 ml, of ether until the base is completely C. In the test for Related substances, the principal spot in the
extracted. Combine the ether extracts, wash with two quantities, chromatogram obtained with test solution (b) corresponds to
each of 10 ml, of water and extract the aqueous washings with that in the chromatogram obtained with reference solution (b).
734
IP 2007 MEPYRAMINE TABLETS
Tests Mepyramine Tablets

Appearance of solution. A 4.0 per cent w/v solution in carbon Mepyramine MaleateTablets; Pyrilamine Maleate
Tablets; Pyrilamine Tablets
Mepyramine Tablets contain not less than 95.0 per cent and
Related substances. Determine by thin-layer chromatography mepyramine maleate, C17H23N3O,C4H4O4. The tablets may be
(2.4.17), coating the plate with silica gel GF254. coated.
Mobile phase. A mixture of 100 volumes of ethyl acetate and
2 volumes of diethylamine. Identification
Prepare the following solutions immediately before use. A. Shake a quantity of the powdered tablets containing 0.1 g
of Mepyramine Maleate with 10 ml of dichloromethane, filter
and evaporate the filtrate to dryness.
obtained with mepyramine maleate RS or with the reference
Reference solution (a). A 4.0 per cent w/v solution of spectrum of mepyramine maleate.
mepyramine maleate RS in chloroform.
Reference solution (b). A 0.4 per cent w/v solution of chromatogram obtained with test solution (b) corresponds to
mepyramine maleate RS in chloroform. that in the chromatogram obtained with reference solution (b).
Reference solution (c). A 0.008 per cent w/v solution of C. Dissolve a quantity of the powdered tablets containing 0.2
mepyramine maleate RS in chloroform. g of Mepyramine Maleate, freed as far as possible from any
Reference solution (d). A 0.004 per cent w/v solution of sugar coating, in 3 ml of water, add 2 ml of 5 M sodium
mepyramine maleate RS in chloroform. hydroxide and shake with three quantities, each of 3 ml, of
Apply to the plate 5 µl of each solution. After development, ether. Warm the aqueous layer in a water-bath for 10 minutes
dry the plate in air and examine in ultraviolet light at 254 nm. with 2 ml of bromine solution, heat to boiling, cool and add
Any secondary spot in the chromatogram obtained with test 0.2 ml to a solution of 10 mg of resorcinol in 3 ml of sulphuric
solution (a) is not more intense than the spot in the acid; a bluish black colour is produced on heating for 15
chromatogram obtained with reference solution (c). The test minutes in a water-bath.
is not valid unless the Rf values of the principal spots in the Tests
chromatograms obtained with test solution (a) and reference
solution (a) are at least 0.2 and unless the spot in the Related substances. Determine by thin-layer chromatography
chromatogram obtained with reference solution (d) is clearly (2.4.17), coating the plate with silica gel GF254.
visible. Ignore the spot due to maleic acid on the line of Mobile phase. A mixture of 100 volumes of ethyl acetate and
application. 2 volumes of diethylamine.
Heavy metals (2.3.13).1.0 g complies with the limit test for Prepare the following solutions immediately before use.
heavy metals, Method A (20 ppm). Test solution (a). Shake a quantity of the powdered tablets
Chlorides (2.3.12). 2.5 g dissolved in 15 ml of water complies containing 0.4 g of Mepyramine Maleate with 10 ml of
with the limit test for chlorides (100 ppm). chloroform and filter.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Test solution (b). Dilute 1 ml of test solution (a) to 10 ml with
Loss on drying (2.4.19). Not more than 0.25 per cent, chloroform.
determined on 2.0 g by drying in an oven at 80°. Reference solution (a). A 4.0 per cent w/v solution of
Assay. Weigh accurately about 0.15 g, dissolve in 40 ml of mepyramine maleate RS in chloroform.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Reference solution (b). A 0.4 per cent w/v solution of
acid, determining the end-point potentiometrically (2.4.25). mepyramine maleate RS in chloroform.
Carry out a blank titration. Reference solution (c). A 0.008 per cent w/v solution of
1 ml of 0.1 M perchloric acid is equivalent to 0.02007 g of mepyramine maleate RS in chloroform.
C17H23N3O,C4H4O4. Reference solution (d). A 0.004 per cent w/v solution of
Storage. Store protected from light and moisture. mepyramine maleate RS in chloroform.
735
MERCAPTOPURINE IP 2007
Apply to the plate 5 µl of each solution. After development, When examined in the range 230 nm to 360 nm (2.4.7), the
dry the plate in air and examine in ultraviolet light at 254 nm. resulting solution shows an absorption maximum only at about
Any secondary spot in the chromatogram obtained with test 325 nm.
solution (a) is not more intense than the spot in the C. Dissolve 20 mg in 20 ml of ethanol (95 per cent) heated to
chromatogram obtained with reference solution (c). The test 60° and add 1 ml of a saturated solution of mercuric acetate in
is not valid unless the Rf values of the principal spots in the ethanol (95 per cent); a white precipitate is produced.
chromatograms obtained with test solutions (a) and reference
solution (a) are at least 0.2 and unless the spot in the D. Dissolve 20 mg in 20 ml of ethanol (95 per cent) heated at
chromatogram obtained with reference solution (d) is clearly 60º and add 1 ml of a 1 per cent w/v solution of lead acetate in
visible. Ignore the spot due to maleic acid on the line of ethanol (95 per cent); a yellow precipitate is produced.
application. Tests
Hypoxanthine. Dissolve 50 mg in 5 ml of dimethyl sulphoxide
Assay. Weigh and powder 20 tablets. Weigh accurately a and add sufficient 0.1 M hydrochloric acid to produce 500
quantity of the powder containing about 0.1 g of Mepyramine ml. Dilute 25 ml to 1000 ml with 0.1 M hydrochloric acid and
Maleate, add 75 ml of water and 5 ml of 2 M hydrochloric measure the absorbance of the resulting solution at about 325
acid, shake vigorously for 15 minutes and dilute to 100.0 ml nm (2.4.7). Dilute a further 50 ml of the original solution to 100
with water. Centrifuge and dilute 10.0 ml of the clear, ml with 0.1 M hydrochloric acid and measure the absorbance
supernatant liquid to 100.0 ml with water. To 10.0 ml add 10 ml of the resulting solution at about 255 nm; the ratio of the
of 0.1 M hydrochloric acid and dilute to 50.0 ml with water. absorbance at about 255 nm to that at about 325 nm, not
Measure the absorbance of the resulting solution at the greater than 1.05.
C17H23N3O,C4H4O4 taking 206 as the specific absorbance at
316 nm. Water (2.3.43). 10.0 to 12.0 per cent, determined on 0.25 g.
Storage. Store protected from light and moisture. Assay. Weigh accurately about 0.15 g, dissolve in 50 ml of
dimethylformamide. Titrate with 0.1 M tetrabutylammonium
Mercaptopurine 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
SH 0.01522 g of C5H4N4S.
H
N Storage. Store protected from light and moisture.
N
N N
Mercaptopurine Tablets
C5H4N4S,H2O Mol. Wt. 170.2
Mercaptopurine Tablets contain not less than 90.0 per cent
Mercaptopurine is purine-6-thiol monohydrate. and not more than 110.0 per cent of the stated amount of
Mercaptopurine contains not less than 98.5 per cent and not mercaptopurine, C5H4N4S,H2O.
more than 101.0 per cent of C5H4N4S, calculated on the
Identification
anhydrous basis.
Description. A yellow, crystalline powder. Shake a quantity of the powdered tablets containing 50 mg of
Mercaptopurine with a mixture of 20 ml of water and 0.5 ml of
Identification 5 M sodium hydroxide for about 3 minutes, add sufficient
water to produce 100 ml, mix and filter. Dilute a suitable aliquot
Test A may be omitted if tests B, C and D are carried out. Tests of the filtrate with sufficient 0.1 M hydrochloric acid to give
B, C and D may be omitted if test A is carried out. a solution containing 5 µg of Mercaptopurine per ml. The
A. Determine by infrared absorption spectrophotometry (2.4.6). resulting solution shows an absorption maximum at about
Compare the spectrum with that obtained with mercaptopurine 325 nm (2.4.7).
RS.
Tests
B. Dissolve 20 mg in 5 ml of dimethyl sulphoxide and add
sufficient 0.1 M hydrochloric acid to produce 100 ml. Dilute Dissolution (2.5.2).
5 ml to 200 ml with 0.1 M hydrochloric acid. Apparatus. No 1
736
IP 2007 MEROPENEM
Medium. 900 ml of 0.1 M hydrochloric acid Meropenam is (1R,5S,6S)-2-[(3S,5S)-5-

Speed and time. 50 rpm and 60 minutes. (dimethylaminocarbonyl)pyrrolidin-3-ylthio-6-[(R)-1-
hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylic acid.
Meropenem contains not less than 98.0 per cent and not more
than 101.0 per cent of C17H 25N3O5S, calculated on the
Test solution. Use the filtrate as the test solution. anhydrous basis.
Reference solution. A solution containing 0.0055 per cent w/v Description. A white to off-white crystalline powder.
of mercaptopurine RS in the mobile phase.
octadecylsilane chemically bonded to porous silica Compare the spectrum with that obtained with meropenem RS
(5 µm), or with the reference spectrum of meropenem.
– mobile phase: 0.1 per cent v/v solution of acetic acid in
water, Tests
– spectrophotometer set at 230 nm, pH (2.4.24). 4.0 to 6.0, determined in 1.0 per cent w/v solution
– a 20 µl loop injector. in water.
Inject the reference solution. The test is not valid unless the Specific optical rotation (2.4.22). –17.0º to -21.0º, determined
retention time for mercaptopurine is not less than 4 minutes in a 0.5 per cent w/v solution.
and the relative standard deviation for replicate injections is Acetone (5.4). Not more than 0.05 per cent.
not more than 2.0 per cent.
Calculate the content of C5H4N4S in the medium. examination in 0.2 ml of dimethylformamide and 2.0 ml of internal
D. Not less than 75 per cent of the stated amount of C5H4N4S. standard solution.
Other tests. Comply with the tests stated under Tablets. Reference solution. Weigh accurately about 50 mg of acetone,
dissolve in a small quantity of dimethylformamide, dilute to
100.0 ml with dimethylformamide and mix. To 1.0 ml of this
solution, add 10.0 ml of the internal standard solution, and
Mercaptopurine, dissolve as completely as possible in 5 ml of
mix.
dimethyl sulphoxide and add sufficient 0.1 M hydrochloric
acid to produce 500.0 ml. Dilute 5.0 ml to 100.0 ml with 0.1 M Internal standard solution. A 0.000005 per cent w/v solution
hydrochloric acid, filter if necessary and measure the of ethyl acetate in dimethylformamide.
absorbance of the resulting solution at the maximum at about Chromatographic system
325 nm (2.4.7). Calculate the content of C5H4N4S, H2O taking – a glass column 2 m × 3 mm, packed with styrenedivinyl-
1165 as the specific absorbance at 325 nm. benzene copolymer (such as Chromosorb 101),
Storage. Store protected from light and moisture. – temperature:
column. 150º,
inlet port and detector 170º,
– flow rate adjusted so that the retention time for acetone
Meropenem is about 3 minutes of the carrier gas.
Inject 1 µl of the test solution and the reference solution.
COOH
O Calculate the percentage of acetone.
N O
HO Related substances. Determine by liquid chromatography
S
N CH3 (2.4.14).
H3C H H NH
CH3 CH3 Solvent mixture. To 900 ml of water add 1.0 ml of triethylamine,
adjust the pH to 5.0 with dilute phosphoric acid, dilute to
C17H25N3O5S, 3H2O Mol. Wt. 437.5 (hydrated) 1000 ml with water and mix.
383.5 (anhydrous) NOTE – Prepare the solutions immediately before use.
737
MEROPENEM INJECTION IP 2007
Test solution. Dissolve 0.5 g of the substance under Inject the reference solution. The test is not valid unless the
examination in the solvent mixture and dilute to 100 ml of the tailing factor is not more than 1.5, the column efficiency is not
solvent mixture. less than 2500 theoretical plates and the relative standard
Reference solution. A 0.0025 per cent w/v solution of deviation for replicate injections is not more than 2.0 per cent.
meropenem RS in the solvent mixture. Inject alternately the test solution and the reference solution.
Chromatographic system Calculate the content of C17H25N3O5S.
octadecylsilane bonded to porous silica (5 µm), Meropenem intended for use in the manufacture of parenteral
– column temperature 40º, preparations without a further appropriate procedure for
– mobile phase: mix 1.0 ml of triethylamine and 900 ml of the removal of bacterial endotoxins complies with the
water, adjust the pH to 5.0 with dilute phosphoric acid, following additional requirement.
dilute with water to 1000 ml, add 70 volumes of Bacterial endotoxins (2.2.3). Not more than 0.125 Endotoxin
acetonitrile and mix, Unit per mg of meropenem.
Meropenem intended for use in the manufacture of parenteral
preparations without a further appropriate sterilisaion
procedure complies with the following additional
Inject the reference solution. The test is not valid unless the requirement.
less than 2500 theoretical plates and the relative standard
deviation for replicate injections is not more than 2.0 per cent. Storage. Store in airtight containers, at a temperature not
Inject the test solution and the reference solution. Any exceeding 25º.
individual impurity is not more than 0.5 per cent and the sum Labelling. The label states whether or not the contents are
of all impurities found is not more than 2.0 per cent. intended for use in the manufacture of parenteral preparations.
Sulphated ash (2.3.18). Not more than 0.1per cent, igniting at Meropenem Injection
500 ± 50º, instead of at 800 ± 25º. Use a desiccator containing
silica gel. Meropenem Injection is a sterile material consisting of
Meropenem and Sodium Carbonate.
Water (2.3.43). 11.4 per cent to 13.4 per cent, determined on
0.5 g. The injection is constituted by dissolving the contents of the
Solvent mixture. To 900 ml of water add 1.0 ml of triethylamine,
adjust the pH to 5.0 with dilute phosphoric acid, dilute to The constituted solution complies with the requirements for
1000 ml with water and mix. Clarity of solution and Particulate matter stated under
Test solution. Dissolve 50.0 mg of the substance under after preparation but, in any case, within the period
examination in the solvent mixture and dilute to 100.0 ml with recommended by the manufacturer.
the solvent mixture.
Meropenem Injection contains not less than 90.0 per cent and
Reference solution. A 0.05 per cent w/v solution of meropenem
meropenem, C17H25N3O5S.
– mobile phase: a mixture of 50 volumes of the solvent Identification
mixture and 10 volumes of methanol,
– flow rate. 1.5 ml per minute, In the Assay, the principal peak in the chromatogram obtained
– spectrophotometer set at 300 nm, with the test solution corresponds to the peak in the
738
IP 2007 MESTRANOL
Tests – mobile phase: dilute 153 volumes of buffer solution

prepared by dissolving 20 ml of 25 per cent w/v of
pH (2.4.24). 7.3 to 8.3, determined in 5.0 per cent w/v solution. tetrabutylammonium hydroxide to 1000 ml with water.
Related substances. Determine by liquid chromatography Adjust the pH to 7.5 with dilute phosphoric acid, add
(2.4.14). 30 volumes of acetonitrile and 20 volumes of methanol
and mix,
Solvent mixture. Dissolve 1.0 ml of triethylamine in 900 ml of
water. Adjust the pH to 5.0 with dilute phosphoric acid and
dilute to 1000 ml with water.
Test solution. Determine the weight of the contents of 10 tailing factor is not more than 1.5, the column efficiency in not
containers. Dissolve an accurately weighed quantity of the less than 2500 theoretical plates and the relative standard
mixed contents of the 10 containers containing about 50 mg of deviation for replicate injections is not more than 2.0 per cent.
Meropenem in 10 ml of the solvent mixture, and mix.
meropenem RS in solvent mixture. Calculate the content of C17H25N3O5S in the injection.
Chromatographic system Storage. Store protected from moisture.

– a stainless steel column 25 cm × 4.0 mm, packed with Labelling. The label states the quantity in mg, of sodium (Na)
octadecylsilane bonded to porous silica (3 µm), in a suitable dose-volume.
– mobile phase: mix 1.0 ml of triethylamine and 900 ml of
water, adjust the pH to 5.0 with dilute phosphoric acid,
dilute to 1000 ml with water, filter and mix with 60 volumes Mestranol
of acetonitrile,
– flow rate. 1 ml per minute, H3C OH
– spectrophotometer set at 220 nm, C CH
– a 10 µl loop injector. H
tailing factor is not more than 1.5 and the column efficiency is H H
not less than 2500 theoretical plates. H3CO
Inject the test solution and the reference solution. Any
C21H26O2 Mol. Wt. 310.4
individual impurity is not more than 0.8 per cent and the sum
of all impurities found is not more than 2.0 per cent. Mestranol is 3-methoxy-19-nor-17α-pregna-1,3,5(10)-trien-
20yn-17β−ol.
Unit per mg of meropenem. Mestranol contains not less than 98.0 per cent and not more
Sterility (2.2.11). Complies with the test for sterility. than 102.0 per cent of C21H26O2, calculated on the dried basis.
Loss on drying (2.4.19). 9.0 per cent to 12.0 per cent, determined Description. A white or almost white, crystalline powder.
on 1.0 g by drying it in vaccume oven at 65º for 6 hours.
Identification
Test B may be omitted if tests A and C are carried out. Test A
Test solution. Determine the weight of the contents of 10 may be omitted if tests B and C are carried out.
containers. Dissolve an accurately weighed quantity of the
mixed contents of the 10 containers containing about 10 mg of A. Determine by infrared absorption spectrophotometry (2.4.6).
Meropenem in the mobile phase and dilute to 100.0 ml with the Compare the spectrum with that obtained with mestranol RS.
mobile phase. B. In the test for Related substances the principal spot in the
Reference solution. A 0.01 per cent w/v solution of meropenem chromatogram obtained with test solution (b) corresponds to
RS in the mobile phase. that in the chromatogram obtained with reference solution (c).
Chromatographic system C. Dissolve about 5 mg in 1 ml of sulphuric acid; a red colour
– a stainless steel column 25 cm × 4.6 mm, packed with is produced which appears greenish-yellow in ultraviolet light
octadecylsilane bonded to porous silica (5 µm), at 365 nm. On adding the solution to 10 ml of water and mixing,
739
METFORMIN HYDROCHLORIDE IP 2007
the solution becomes pink and on standing a pink to violet Metformin Hydrochloride
precipitate is produced.
Tests CH3 H
N N NH2 , HCl
Specific optical rotation (2.4.22). –20.0° to – 24.0°, determined H3C
in a 1.0 per cent w/v solution in anhydrous pyridine. NH NH
Light absorption (2.4.7). Dissolve about 25 mg in sufficient C4H11N5,HCl Mol. Wt. 165.6
ethanol (95 per cent) to produce 25 ml and dilute 10 ml of the
solution to 100 ml with ethanol (95 per cent). When examined Metformin Hydrochloride is 1,1-dimethylbiguanide
in the range 230 nm to 360 nm, the resulting solution shows hydrochloride.
absorption maxima at about 279 nm and 288 nm and a minimum Metformin Hydrochloride contains not less than 98.5 per cent
at about 286 nm. Absorbance at about 279 nm is 0.062 to 0.068 and not more than 101.0 per cent of C4H11N5,HCl, calculated
and at about 288 nm is 0.059 to 0.064, both calculated on the on the dried basis.
dried basis.
Description. A white, crystalline powder; hygroscopic.
(2.4.17), coating the plate with silica gel G. Identification
Mobile phase. A mixture of 90 volumes of toluene and 10 A. Determine by infrared absorption spectrophotometry (2.4.6).
volumes of ethanol (95 per cent). Compare the spectrum with that obtained with metformin
Test solution (a). Dissolve 0.1 g of the substance under hydrochloride RS or with the reference spectrum of metformin
examination in 10 ml of chloroform. hydrochloride.
Test solution (b). Dissolve 0.1 g of the substance under B. Dissolve 25 mg in 5 ml of water, add 1.5 ml of 5 M sodium
examination in 100 ml of chloroform. hydroxide, 1 ml of 1-naphthol solution and, dropwise with
shaking, 0.5 ml of dilute sodium hypochlorite solution; an
orange-red colour is produced which darkens on keeping.
substance under examination in chloroform.
Reference solution (b). A 0.005 per cent w/v solution of the C. Dissolve 10 mg in 10 ml of water and add 10 ml of a solution
substance under examination in chloroform. prepared by mixing equal volumes of a 10 per cent w/v solution
of sodium nitroprusside, a 10 per cent w/v solution of
Reference solution (c). A 0.1 per cent w/v solution of potassium ferricyanide and a 10 per cent w/v solution of
mestranol RS in chloroform. sodium hydroxide and allowing to stand for 20 minutes; a
Apply to the plate 5 µl of each solution. After development, wine red colour develops within 3 minutes.
dry the plate in air until the odour of the solvent is no longer D. Gives reaction A of chlorides (2.3.1).
detectable, heat it at 110° for 10 minutes and spray the hot
plate with ethanolic sulphuric acid (20 per cent). Heat again Tests
at 110° for 10 minutes and examine in daylight and in ultraviolet
light at 365 nm. Any secondary spot in the chromatogram Related substances. Determine by liquid chromatography
obtained with test solution (a) is not more intense than the (2.4.14).
spot in the chromatogram obtained with reference solution Test solution. Dissolve 50 mg of the substance under
(a), and not more than one such spot is more intense than the examination in 10 ml of water.
(b). Reference solution (a). A 0.0005 per cent w/v solution of the
substance under examination in water.
on 0.5 g by drying in an oven at 105° for 3 hours. Reference solution (b). A 0.0001 per cent w/v solution of
dicyandiamide in water.
tetrahydrofuran and add 5 ml of 10 per cent w/v solution of Chromatographic system
silver nitrate. Titrate with 0.1 M sodium hydroxide, – a stainless steel column 30 cm x 4 mm, packed with
determining the end-point potentiometrically (2.4.25). octadecylsilane bonded to porous silica (10 µm),
– mobile phase: a solution containing 0.087 per cent w/v
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03104 g of of sodium pentanesulphonate and 0.12 per cent w/v of
C21H26O2. sodium chloride, adjusted to pH 3.5 using 1 per cent v/
Storage. Store protected from light and moisture. v solution of orthophosphoric acid,
740
IP 2007 METFORMIN TABLETS
– flow rate. 1 ml per minute, Tests

– a 20 µl loop injector. Related substances. Determine by liquid chromatography
(2.4.14).
For the test solution record the chromatogram for three times
the retention time of the principal peak. In the chromatogram Test solution. Shake a quantity of the powdered tablets
obtained with the test solution the area of any peak containing 0.5 g of Metformin Hydrochloride with 100 ml of
corresponding to dicyandiamide is not greater than that water and filter.
obtained with reference solution (b) and the area of any other Reference solution (a). Dilute 0.1 ml of the test solution to
secondary peak is not greater than that obtained with reference 100 ml with water.
solution (a).
Heavy metals (2.3.13). 1.0 g complies with the limit test for dicyandiamide in water.
Sulphated ash (2.3.18). Not more than 0.1 per cent. – a stainless steel column 30 cm x 4 mm, packed with
Loss on drying (2.4.19). Not more than 0.5 per cent, determined octadecylsilane bonded to porous silica (10 µm),
on 1.0 g by drying in an oven at 105°. – mobile phase: a solution containing 0.087 per cent w/v
of sodium pentanesulphonate and 0.12 per cent w/v of
Assay. Weigh accurately about 60 mg, dissolve in 4 ml of
sodium chloride, adjusted to pH 3.5 using 1 per cent
anhydrous formic acid, add 50 ml of acetic anhydride. Titrate
v/v solution of orthophosphoric acid,
with 0.1 M perchloric acid, determining the end-point
1 ml of 0.1 M perchloric acid is equivalent to 0.008281 g of – a 20 µl loop injector.
C4H11N5,HCl.
For the test solution record the chromatogram for three times
Storage. Store protected from light and moisture. the retention time of the principal peak. In the chromatogram
obtained with the test solution the area of any peak
corresponding to dicyandiamide is not greater than that
Metformin Tablets obtained with reference solution (b) and the area of any other
secondary peak is not greater than that obtained with reference
Metformin Hydrochloride Tablets solution (a).
Metformin Tablets contain not less than 95.0 per cent and not Dissolution (2.5.2).
more than 105.0 per cent of the stated amount of metformin Apparatus. No 2
hydrochloride, C4H11N5,HCl. The tablets may be coated.
Medium. 900 ml of a 0.68 per cent w/v solution of potassium
Identification dihydrogen phosphate, adjusted to pH 6.8 by the addition of
1 M sodium hydroxide
A. Shake a quantity of the powdered tablets containing 20 mg Speed and time. 100 rpm and 45 minutes.
of Metformin Hydrochloride with 20 ml of ethanol, filter,
evaporate the filtrate to dryness on a water-bath and dry the Withdraw a suitable volume of the medium and filter, dilute
residue at 105° for 1 hour. suitably with water and measure the absorbance of the
On the residue, determine by infrared absorption Calculate the content of C4H11N5, HCl, in the medium taking
spectrophotometry (2.4.6). Compare the spectrum with that 806 as the specific absorbance at 233 nm.
obtained with metformin hydrochloride RS or with the
reference spectrum of metformin hydrochloride. D. Not less than 70 per cent of the stated amount of C4H11N5,
HCl.
B.Triturate a quantity of the powdered tablets containing
50 mg of Metformin Hydrochloride with 10 ml of water and Other tests. Comply with the tests stated under Tablets.
filter. To 5 ml of the filtrate, add 1.5 ml of 5 M sodium hydroxide,
1 ml of 1-naphthol solution and, dropwise with shaking, 0.5
quantity of the powder containing about 0.1 g of Metformin
ml of dilute sodium hypochlorite solution; an orange-red
Hydrochloride, shake with 70 ml of water for 15 minutes, dilute
colour is produced which darkens on keeping.
to 100.0 ml with water and filter. Dilute 10.0 ml of the filtrate to
C.The filtrate obtained in test B gives reaction A of chlorides 100.0 ml with water. Further dilute 10.0 ml to 100.0 ml with
(2.3.1). water and measure the absorbance of the resulting solution
741
METHADONE HYDROCHLORIDE IP 2007
at the maximum at about 232 nm (2.4.7). Calculate the content Related substances. Determine by thin-layer chromatography
of C4H11N5, HCl taking 798 as the specific absorbance at 232 (2.4.17), coating the plate with silica gel G.
nm. Mobile phase. A mixture of 60 volumes of ethanol (95 per
cent), 30 volumes of glacial acetic acid and 10 volumes of
water.
Methadone Hydrochloride Test solution. Dissolve 0.5 g of the substance under
Amidone Hydrochloride examination in 10 ml of ethanol (95 per cent).
CH3
O CH3 substance under examination in ethanol (95 per cent).
N Apply to the plate 10 µl of each solution. After development,
CH3 , HCl dry the plate in air, spray with dilute potassium
CH3 iodobismuthate solution. Any secondary spot in the
reference solution.
C21H27NO,HCl Mol. Wt. 345.9
Methadone Hydrochloride is (RS)-dimethyl-(1-methyl-4-
oxo-3,3-diphenylhexyl)amine hydrochloride. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Methadone Hydrochloride contains not less than 98.5 per
cent and not more than 100.5 per cent of C21H27NO,HCl, Assay. Weigh accurately about 0.5 g, dissolve in 50 ml of
calculated on the dried basis. anhydrous glacial acetic acid, add 5 ml of mercuric acetate
solution. Titrate with 0.1 M perchloric acid, using crystal
Description. A white, crystalline powder. violet solution as indicator and continuing the titration until
Identification the colour changes from violet-blue to green. Carry out a blank
titration.
C21H27NO,HCl.
Compare the spectrum with that obtained with methadone
hydrochloride RS.
B. To 2 ml of a 5 per cent w/v solution in carbon dioxide-free
water add 1 ml of 0.1 M hydrochloric acid and 6 ml of Methadone Injection
ammonium thiocyanate solution; a white precipitate is
Methadone Injection; Amidone Hydrochloride Injection;
produced which becomes crystalline on stirring for a few
Amidone Injection
minutes. The precipitate, after drying at 105° melts at 143° to
148° (2.4.21). Methadone Injection is a sterile solution of Methadone
C. Dissolve 50 mg in 5 ml of carbon dioxide-free water, add 1
ml of 6 M ammonia, mix, allow to stand for 5 minutes and filter; Methadone Injection contains not less than 92.5 per cent and
the filtrate gives reaction A of chlorides (2.3.1). not more than 107.5 per cent of the stated amount of
methadone hydrochloride, C21H27NO,HCl.
D. Optical rotation of a 2-dm layer of a 5 per cent w/v solution
in carbon dioxide-free water, is – 0.05° to +0.05° (2.4.22). Identification
Tests Make a volume containing 0.1 g of Methadone Hydrochloride
Appearance of solution. A 5.0 per cent w/v solution in carbon alkaline with 5 M sodium hydroxide, stir with a glass rod until
dioxide-free water is clear (2.4.1), and colourless (2.4.1). the precipitate solidifies, filter, wash with water and dry over
phosphorus pentoxide at room temperature at a pressure of
Acidity or alkalinity. To 10 ml of a 2.0 per cent w/v solution in 2 kPa. The residue complies with the following tests.
carbon dioxide-free water add 0.2 ml of methyl red solution
and 0.2 ml of 0.01 M sodium hydroxide; the solution is yellow. A. Determine by infrared absorption spectrophotometry (2.4.6).
Add 0.4 ml of 0.01 M hydrochloric acid; the solution is red. Compare the spectrum with that obtained with methadone
742
IP 2007 METHDILAZINE HYDROCHLORIDE
hydrochloride RS treated in the same manner or with the stand for 2 hours The residue, after recrystallisation from
reference spectrum of methadone. ethanol (20 per cent), washing with ethanol (20 per cent)
B. To 5 mg add 0.05 ml of dinitrobenzene solution and 0.05 ml and drying at 105°, melts at about 160° or 180° (2.4.21).
of a 50 per cent w/v solution of sodium hydroxide; a purple Tests
colour is produced which changes slowly to dark brown.
Tests Tablets.
pH (2.4.24). 5.0 to 6.5. Crush one tablet to a fine powder and transfer to a 25-ml
Other tests. Complies with the tests stated under Parenteral volumetric flask. Add 20 ml of water, mix by shaking and dilute
Preparations (Injections). to volume with water. Mix well and centrifuge. Dilute a suitable
volume of the clear, supernatant liquid with water to produce
a solution containing about 0.2 mg of Methadone
10 mg of Methadone Hydrochloride add 1 ml of glacial acetic
Hydrochloride per ml. Measure the absorbance of the resulting
acid and dilute to 100.0 ml with water. To 10.0 ml of this solution
add 10 ml of a 0.4 per cent w/v solution of picric acid and
content of C21H27NO,HCl in the tablet from the absorbance
10 ml of phosphate buffer pH 4.9, extract with three quantities,
obtained by repeating the operation on an accurately weighed
each of 15 ml, of chloroform, dilute the combined chloroform
quantity of methadone hydrochloride RS.
extracts to 50.0 ml with chloroform. To 10.0 ml add sufficient
chloroform to produce 20.0 ml and measure the absorbance of Other tests. Comply with the tests stated under Tablets.
the resulting solution at the maximum at about 350 nm (2.4.7), Assay. Weigh and powder 20 tablets. Weigh accurately a
using as the blank a solution prepared in the same manner but quantity of the powder containing about 50 mg of Methadone
omitting the substance under examination. Calculate the Hydrochloride, add 60 ml of water and 5 ml of glacial acetic
content of C21H27NO,HCl taking 448 as the specific absorbance acid, heat on a water-bath for 5 minutes, mix with the aid of
at 350 nm. ultrasound for 10 minutes and dilute to 100.0 ml with water.
Storage. Store protected from light, in single dose container. Filter, discarding the first 10 ml of the filtrate, and dilute 20.0 ml
to 100.0 ml with water. To 10.0 ml of the resulting solution, add
10 ml of a 0.4 per cent w/v solution of picric acid and 10 ml of
Methadone Tablets phosphate buffer pH 4.9, extract with three quantities, each of
15 ml, of chloroform, dilute the combined chloroform extracts
Methadone Tablets; Amidone Hydrochloride Tablets; to 50.0 ml with chloroform. To 10.0 ml add sufficient chloroform
Amidone Tablets to produce 20.0 ml and measure the absorbance of the resulting
Methadone Tablets contain not less than 92.5 per cent and
blank a solution prepared in the same manner but omitting the
substance under examination. Calculate the content of
methadone hydrochloride, C21H27NO,HCl.
C21H27NO,HCl taking 448 as the specific absorbance at
Identification 350 nm.
A. Shake a quantity of the powdered tablets containing 0.1 g Storage. Store protected from light and moisture.
of Methadone Hydrochloride with 20 ml of water and
centrifuge. Make the supernatant liquid alkaline with 5 M
sodium hydroxide, stir with a glass rod until the precipitate Methdilazine Hydrochloride
solidifies, filter, wash with water and dry over phosphorus
pentoxide at room temperature at a pressure of 2 kPa.
On the residue, determine by infrared absorption N CH3
obtained with methadone hydrochloride RS or with the N , HCl
reference spectrum of methadone.
B. Extract a quantity of the powdered tablets containing 0.1 g S
of Methadone Hydrochloride with 10 ml of water, filter and
C18H20N2S,HCl Mol. Wt. 332.9
wash the residue with sufficient water to bring the volume of
the filtrate to 10 ml. Add to the filtrate 0.125 g of picrolonic Methdilazine Hydrochloride is 10-(1-methylpyrrolidin-3-
acid dissolved in 50 ml of boiling water, stir and allow to ylmethyl)phenothiazine hydrochloride.
743
METHDILAZINE TABLETS IP 2007
Methdilazine Hydrochloride contains not less than 97.0 per Identification

cent and not more than 103.0 per cent of C18H20N2S,HCl,
calculated on the dried basis. Weigh accurately a quantity of finely powdered tablets
containing about 8 mg of Methdilazine Hydrochloride, transfer
Description. A light tan powder, turning pale pink to brown to a 60-ml separating funnel, add 10 ml of a 10 per cent w/v
on exposure to light; odour, slight and characteristic. solution of sodium bicarbonate and extract with 3 ml of
chloroform. Filter the extract through a plug of cotton.
Identification
Evaporate the chloroform carefully removing the last traces
A. Determine by infrared absorption spectrophotometry (2.4.6). of solvent in a small flask at a pressure of 1.5 to 2.5 kPa.
Compare the spectrum with that obtained with methdilazine On the residue, determine by infrared absorption
hydrochloride RS or with the reference spectrum of spectrophotometry (2.4.6). Compare the spectrum with that
methdilazine hydrochloride. obtained with methdilazine hydrochloride RS or with the
B. When examined in the range 230 nm to 360 nm (2.4.7), a reference spectrum of methdilazine hydrochloride.
0.0005 per cent w/v solution shows an absorption maximum at
about 252 nm and an inflection at about 275 nm; absorbance Tests
at about 252 nm, about 0.46.
C. Dissolve 50 mg in dilute hydrochloric acid, add 3 ml of Tablets.
buffered palladium chloride solution and 1 ml of a 1 per cent
w/v solution of sodium lauryl sulphate and mix; a dark blue Carry out the procedure protected from light.
colour is formed. Powder one tablet and dissolve in about 20 ml of methanol in
D. Gives the reactions of chlorides (2.3.1). a 25-ml volumetric flask, dilute to volume and mix. Filter,
discarding the first 10 ml of the filtrate. To 10.0 ml of the filtrate
Tests in a 100-ml volumetric flask, add 5 ml of methanol and 4.0 ml of
buffered palladium chloride solution, dilute to volume with
ethanol (95 per cent) and mix. Carry out the Assay on the
Heavy metals (2.3.13).1.0 g complies with the limit test for resulting solution beginning at the words “Measure the
heavy metals, Method B (20 ppm). absorbance.....”. Calculate the content of C18H20N2S, HCl in
Sulphated ash (2.3.18). Not more than 0.5 per cent. the tablet.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined Other tests. Comply with the tests stated under Tablets.
on 1.0 g, by drying in an oven at 65° at a pressure not exceeding Assay. Carry out the procedure protected from light.
the powder containing about 80 mg of Methdilazine
water to produce 100.0 ml. Transfer 5.0 ml of this solution to a
Hydrochloride and transfer to a 200-ml volumetric flask. Add
100-ml volumetric flask, dilute to volume with water and mix.
60 ml of methanol, shake for 20 minutes, dilute with methanol
Dilute 5.0 ml to 50.0 ml with water, mix and measure the
to volume and mix. Filter, discarding the first 15 ml of the
filtrate. To 10.0 ml of the filtrate in a 100-ml volumetric flask
252 nm and at about 275 nm (2.4.7). Subtract the absorbance at
add 20 ml of methanol and 4.0 ml of buffered palladium
about 275 nm from the absorbance at about 252 nm. Calculate
chloride solution, dilute to volume with ethanol (95 per cent)
the content of C18H20N2S,HCl from the difference in the
and mix. Measure the absorbance of the resulting solution at
absorbances obtained by carrying out the Assay
the maximum at about 460 nm (2.4.7), using as the blank a
simultaneously on methdilazine hydrochloride RS.
solution prepared by treating 10 ml of methanol in the same
Storage. Store protected from light and moisture. manner but omitting the filtrate of methdilazine hydrochloride
solution. Calculate the content of C18H20N2S,HCl from the
absorbance obtained by repeating the operation using
methdilazine hydrochloride RS instead of the substance under
Methdilazine Tablets examination.
Methdilazine Hydrochloride Tablets Storage. Store protected from light and moisture.
Methdilazine Tablets contain not less than 90.0 per cent and
methdilazine hydrochloride, C18H20N2S,HCl.
744
IP 2007 METHOTREXATE INJECTION
Methotrexate – mobile phase: a mixture of 92 volumes of phosphate

COOH – spectrophotometer set at 302 nm,
HOOC NH
NH2 O relative standard deviation for replicate injections is not more
N than 2.0 per cent.
N N
Inject reference solution (b). The resolution between the peaks
CH3 due to methotrexate and folic acid is not less than 5.0.
H2N N N
Inject separately the test solution and reference solution (a)
C20H22N8O5 Mol. Wt. 454.4 and measure the responses for the principal peak.
Methotrexate is 4-amino-4-deoxy-10-methylpteroyl-L- Calculate the content of C20H22N8O5.
glutamic acid.
Methotrexate contains not less than 97.0 per cent and not
CAUTION - Great care should be taken to prevent inhaling
more than 102.0 per cent of C20H22N8O5, calculated on the
particles of Methotrexate and exposing the skin to it.
anhydrous basis.
Description. A yellow to orange-brown, crystalline powder.
Identification
Methotrexate Injection
Compare the spectrum with that obtained with methotrexate Methotrexate Injection is a sterile solution of Methotrexate in
RS or with the reference spectrum of methotrexate. Water for Injections containing Sodium Hydroxide.
B. When examined in the range 230 nm to 380 nm (2.4.7), a Methotrexate Injection contains not less than 95.0 per cent
0.001 per cent w/v solution in 0.1 M sodium hydroxide shows and not more than 110.0 per cent of the stated amount of
absorption maxima at about 258 nm, 303 nm and 371 nm; ratio methotrexate, C20H22N8O5.
of the absorbance at the maximum at about 303 nm to that at Description. A clear, yellowish solution.
the maximum at about 371 nm, 2.8 to 3.3.
Identification
Tests
When examined in the range 230 nm to 360 nm (2.4.7), the
Specific optical rotation (2.4.22). +19.0° to +24.0°, determined solution obtained in the Assay shows absorption maxima at
in a 1.0 per cent w/v solution in a solution containing 1.4 per about 242 nm and 306 nm.
cent w/v of sodium carbonate.
Water (2.3.43). Not more than 12.0 per cent, determined on pH (2.4.24). 7.5 to 9.0.
0.25 g.
Assay. Determine by liquid chromatography (2.4.14). Preparations (Injections).
Test solution. Dissolve 25 mg of the substance under Assay. Determine by liquid chromatography (2.4.14).
examination in 250.0 ml of the mobile phase.
Test solution. Dilute a volume of the injection with the mobile
Reference solution (a). Dissolve 25 mg of methotrexate RS in phase to produce a solution containing 0.0025 per cent w/v of
250.0 ml of the mobile phase. Methotrexate.
Reference solution (b). Dissolve 25 mg of methotrexate RS Reference solution (a). A 0.0025 per cent w/v solution of
and 25 mg of folic acid in 250.0 ml of the mobile phase. methotrexate RS in the mobile phase.
Chromatographic system Reference solution (b). A solution containing 0.0025 per cent
– a stainless steel column 25 cm x 4.6 mm, packed with w/v each of methotrexate RS and folic acid in the mobile
octadecylsilane bonded to porous silica (5 µm), phase.
745
METHOTREXATE TABLETS IP 2007
Chromatographic system 306 nm (2.4.7). Calculate the content of C20H22N8O5 taking 430
– a stainless steel column 25 cm x 4.6 mm, packed with as the specific absorbance at 306 nm.
– mobile phase: 92 volumes of phosphate buffer pH 6.0
C20H22N8O5.
and 8 volumes of acetonitrile,
– flow rate. 1.4 ml per minute, Uniformity of content. Comply with the test stated under
– spectrophotometer set at 302 nm, Tablets.
Carry out the test as described under Assay, using the following
Inject reference solution (a). The test is not valid unless the solutions.
relative standard deviation for the replicate injections is not Test solution. Crush one tablet and mix with 100 ml of the
more than 2.0 per cent. mobile phase with the aid of ultrasound, centrifuge and use
Inject reference solution (b). The resolution between the peaks the supernatant liquid.
due to methotrexate and folic acid is not less than 5.0. Reference solution (a). A 0.0025 per cent w/v solution of
Inject separately the test solution and reference solution (a) methotrexate RS in the mobile phase.
and measure the responses for the principal peak. Reference solution (b). A solution containing 0.0025 per cent
Calculate the content of C20H22N8O5 in the injection. w/v each of methotrexate RS and folic acid in the mobile
phase.
Labelling. The label states that the injection is not intended
for intrathecal injection when an antimicrobial preservative is
present. Assay. Determine by liquid chromatography (2.4.14).
a quantity of the powder containing about 2.5 mg of
Methotrexate and mix with 100.0 ml of the mobile phase with
Methotrexate Tablets the aid of ultrasound, centrifuge and use the supernatant liquid.
Methotrexate Tablets contain not less than 90.0 per cent and Reference solution (a). A solution containing 0.0025 per cent
not more than 110.0 per cent of the stated amount of w/v each of methotrexate RS and folic acid in the mobile
methotrexate, C20H22N8O5. phase.
Identification methotrexate RS in the mobile phase.
Extract a quantity of the powdered tablets containing 10 mg of Chromatographic system
Methotrexate with sufficient 0.1 M sodium hydroxide to – a stainless steel column 25 cm x 4.6 mm, packed with
produce 100 ml, filter and dilute 10 ml of the filtrate to 100 ml octadecylsilane bonded to porous silica (5 µm),
with 0.1 M sodium hydroxide. – mobile phase: a mixture of 92 volumes of phosphate
When examined in the range 230 nm to 380 nm (2.4.7), the – flow rate. 1.4 ml per minute,
resulting solution shows absorption maxima at about 258 nm, – spectrophotometer set at 302 nm,
303 nm and 371 nm. – a 20 µl loop injector.
Tests Inject reference solution (a). The test is not valid unless the
Dissolution (2.5.2). than 2.0 per cent.
Apparatus. No 1 Inject reference solution (b). The resolution between the peaks
Medium. 900 ml of 0.1 M hydrochloric acid. due to methotrexate and folic acid is not less than 5.0.
Speed and time. 50 rpm and 45 minutes. Inject separately the test solution and reference solution (a)
Withdraw a suitable volume of the medium and filter promptly and measure the responses for the principal peak.
Calculate the content of C20H22N8O5 in the tablets.
Measure the absorbance of the filtrate at the maximum at about Storage. Store protected from light and moisture.
746
IP 2007 METHOXAMINE INJECTION
Methoxamine Hydrochloride Any spot corresponding to 2,5-dimethoxybenzaldehyde in

OH intense than the spot in the chromatogram obtained with
reference solution (b). Spray the plate with a 0.3 per cent w/v
H3CO CH3
, HCl solution of ninhydrin in 1-butanol containing 3 per cent v/v
NH2 of glacial acetic acid and heat at 105° for 5 minutes. Any
OCH3 other secondary spot in the chromatogram obtained with the
C11H17N03,HCl Mol. Wt. 247.7 test solution is not more intense than the spot in the
Methoxamine Hydrochloride is all-rac-2-amino-1-(2,5-
dimethoxyphenyl)propan-1-ol hydrochloride. Sulphated ash (2.3.18). Not more than 0.2 per cent.
Methoxamine Hydrochloride contains not less than 98.5 per Loss on drying (2.4.19). Not more than 1.0 per cent, determined
cent and not more than 101.0 per cent of C11H17NO3,HCl, on 1.0 g by drying in an oven at 105° for 2 hours.
calculated on the dried basis. Assay. Weigh accurately about 0.5 g and dissolve in 30 ml of
Description. Colourless crystals or white, plate-like crystals anhydrous glacial acetic acid, 15 ml of mercuric acetate
or a white, crystalline powder; odourless or almost odourless. solution and 5 ml of acetic anhydride, warming if necessary.
Titrate with 0.1 M perchloric acid, using crystal violet
Identification solution as indicator. Carry out a blank titration.
Compare the spectrum with that obtained with methoxamine C11H17NO3,HCl.
hydrochloride RS or with the reference spectrum of Storage. Store protected from light and moisture.
methoxamone hydrochloride.
0.005 per cent w/v solution shows an absorption maximum
only at about 290 nm; absorbance at about 290 nm, about 0.69.
Methoxamine Injection
C. Dissolve 20 mg in 2 ml of water, add 5 ml of diazotised Methoxamine Hydrochloride Injection
nitroaniline solution and 1 ml of dilute sodium carbonate Methoxamine Injection is a sterile solution containing 2 per
solution. Allow to stand for 2 minutes and add 1 ml of 1 M cent w/v of Methoxamine Hydrochloride in Water for
sodium hydroxide; a deep red colour is produced which is Injections.
extractable with 1-butanol.
Methoxamine Injection contains not less than 1.90 per cent
D. A 5 per cent w/v solution gives the reactions of chlorides and not more than 2.10 per cent w/v of methoxamine
(2.3.1). hydrochloride, C11H17NO3, HCl.
pH (2.4.24). 4.0 to 6.0, determined in a 2.0 per cent w/v solution. A. When examined in the range 230 nm to 360 nm (2.4.7), the
Related substances. Determine by thin-layer chromatography solution obtained in the Assay shows an absorption maximum
(2.4.14), coating the plate with silica gel GF254. only at about 290 nm.
Mobile phase. A mixture of 86 volumes of chloroform, 12 B. Dilute 1 ml with 1 ml of water, add 5 ml of diazotised
volumes of methanol and 2 volumes of strong ammonia nitroaniline solution and 1 ml of dilute sodium carbonate
solution. solution. Allow to stand for 2 minutes and add 1 ml of 1 M
sodium hydroxide; a deep red colour is produced which is
extractable with 1-butanol.
Reference solution (a). A 0.02 per cent w/v solution of the Tests
Reference solution (b). A 0.01 per cent w/v solution of 2,5- (2.4.17), coating the plate with silica gel GF254.
dimethoxybenzaldehyde in methanol.
Apply to the plate 5 µl of each solution. After development, volumes of methanol and 2 volumes of strong ammonia
dry the plate in air and examine in ultraviolet light at 365 nm. solution.
747
METHYL SALICYLATE IP 2007
Test solution. Dilute the injection, if necessary, with water so ferric chloride test solution; a violet colour develops.
as to contain 2.0 per cent w/v of Methoxamine Hydrochloride. B. Heat 0.25 ml with 2 ml of 2 M sodium hydroxide on a water-
Reference solution (a). Dilute 1 volume of the test solution to bath for 5 minutes and add 3 ml of 1 M sulphuric acid. Filter
100 ml with water. and wash the precipitate with water. The precipitate after
drying at 105° for 1 hour melts at 156° to 161° (2.4.21).
Reference solution (b). A 0.01 per cent w/v solution of 2,5-
dimethoxybenzaldehyde in methanol. Tests
dry the plate in air and examine in ultraviolet light at 365 nm. Appearance of solution. To 2 ml add 10 ml of ethanol (95 per
Any spot corresponding to 2,5-dimethoxybenzaldehyde in cent). The resulting solution is clear (2.4.1), and not more
the chromatogram obtained with the test solution is not more intensely coloured than reference solution YS7 (2.4.1).
intense than the spot in the chromatogram obtained with Acidity. Dissolve 5.0 g in 50 ml of ethanol (95 per cent),
reference solution (b). Spray the plate with a 0.3 per cent w/v previously neutralised to a blue colour with bromocresol green
solution of ninhydrin in 1-butanol containing 3 per cent v/v solution by the addition of 0.1 M sodium hydroxide. Not
of glacial acetic acid and heat at 105° for 5 minutes. Any more than 0.4 ml of 0.1 M sodium hydroxide is required to
other secondary spot in the chromatogram obtained with the restore the blue colour.
Refractive index (2.4.27). 1.534 to 1.538.
Preparations (Injections). Assay. Weigh accurately about 0.5 g, dissolve in 25 ml of
ethanol (95 per cent), add 0.05 ml of phenol red solution and
neutralise with 0.1 M sodium hydroxide. Add 50.0 ml of 0.1 M
100 mg of Methoxamine Hydrochloride add sufficient water
sodium hydroxide and heat under a reflux condenser on a
to produce 100.0 ml. Dilute 5.0 ml of this solution to 100.0 ml
water-bath for 30 minutes. Cool and titrate with 0.1 M
with water. Measure the absorbance of the resulting solution
hydrochloric acid. Carry out a blank titration.
of C11H17NO3,HCl taking 137 as the specific absorbance at 1 ml of 0.1 M sodium hydroxide is equivalent to 0.01522 g of
290 nm. C8H8O3.
Methyl Salicylate Industrial Methylated Spirit

Wintergreen Oil IMS
Industrial Methylated Spirit is a mixture of nineteen volumes
of Ethanol of an appropriate strength and one volume of
COOCH3 approved wood naphtha.
OH Description. Clear, colourless, mobile, volatile liquid; odour,
spirituous and of wood naphtha.
Identification
C8H8O3 Mol. Wt. 152.2
Mix 0.1 ml with 0.05 ml of an 11 per cent w/w solution of
Methyl Salicylate is 2-hydroxybenzoic acid methyl ester. phosphoric acid and 0.25 ml of dilute potassium
Methyl Salicylate contains not less than 99.0 per cent w/w permanganate solution. After 1 minute add a few mg of
and not more than 100.5 per cent w/w of C8H8O3. sodium metabisulphite and shake until the mixture is
decolorised. Add 1.5 ml of a 50 per cent v/v solution of
Description. A colourless or slightly yellow liquid; odour,
sulphuric acid and a few mg of finely powdered chromotropic
strong, persistent, characteristic and aromatic.
acid sodium salt, shake well and heat on a water-bath for 5
Identification minutes; a deep violet colour is produced.
A. To 10 ml of a saturated aqueous solution add 0.05 ml of Tests
748
IP 2007 METHYLCELLULOSE
Relative density (2.4.29). Not greater than 0.815. white, flocculent precipitate is produced which dissolves in
Acidity or alkalinity. 25 ml requires not more than 0.2 ml of 0.1 6 M ammonia.
M sodium hydroxide to produce a pink colour with C. Without heating completely dissolve 0.2 g in 15 ml of a 70
phenolphthalein solution and not more than 1.0 ml of 0.1 M per cent w/w solution of sulphuric acid, pour the solution
hydrochloric acid is required to produce a red colour with with stirring into 100 ml of iced water. In a test-tube kept in ice,
methyl red solution. mix thoroughly 1 ml of the solution with 8 ml of sulphuric
acid, added dropwise. Heat in a water-bath for exactly 3 minutes
Appearance of solution. Dilute 5.0 ml to 100 ml with water; the
and cool immediately in ice. When the mixture is cool, carefully
solution is clear (2.4.1).
add 0.6 ml of a solution containing 3 g of ninhydrin in 100 ml
Aldehydes. Not more than 50 ppm, determined by the following of a 4.55 per cent w/v solution of sodium metabisulphite, mix
method. To 5.0 ml add 5 ml of water and 1 ml of decolorised well and allow to stand at 25°; a pink colour is produced
fuchsin solution and allow to stand for 30 minutes. Any colour immediately which becomes violet within 100 minutes.
produced is not more intense than that obtained by treating in
D. Place 1 ml of solution A on a glass plate. After evaporation
the same manner 5 ml of a 0.005 per cent w/v solution of
of the water a thin film is produced.
redistilled acetaldehyde in aldehyde-free ethanol (95 per
cent). Tests
Non-volatile matter. When evaporated and dried at 105°, leaves
not more than 0.01 per cent w/v of residue. Appearance of solution. Whilst stirring, introduce a quantity
containing 1.0 g of the dried substance into 50 g of carbon
Storage. Store in tightly-closed containers at a temperature dioxide-free water heated to 90°. Allow to cool, dilute to 100 g
not exceeding 30°. with the same solvent and continue stirring until solution is
Labelling. The label states that it is inflammable. complete. Allow to stand at 2° to 8° for 1 hour. The resulting
solution is not more opalescent than opalescence standard
OS3 (2.4.1), and is not more intensely coloured than reference
solution YS6 (2.4.1).
Methylcellulose pH (2.4.24). 5.0 to 8.5, determined in solution A.
Cellulose Methyl Ether Heavy metals (2.3.13).1.0 g complies with the limit test for
Methylcellulose is a cellulose having some of the hydroxyl heavy metals, Method B (20 ppm).
groups in the form of the methyl ether. Various grades are Chlorides (2.3.12). Dilute 5.0 ml of solution A to 15 ml with
available and are distinguished by a number indicative of the water. The resulting solution complies with the limit test for
apparent viscosity in millipascal seconds of a 2 per cent w/w chlorides (0.5 per cent).
solution measured at 20°. Apparent viscosity. Not less than 75 per cent and not more
Methylcellulose contains not less than 27.5 per cent and not than 140 per cent of the declared value, determined by the
more than 31.5 per cent of methoxyl (-OCH3) groups, calculated following method. To 150 g of water heated to 90° add, with
on the dried basis. stirring, a quantity containing 6.0 g of the dried substance.
Stir with a propeller-type stirrer for 10 minutes, place the flask
Description. A white or yellowish white or greyish white
in a bath of iced water, continue the stirring and allow to
powder or granules; practically odourless; hygroscopic after
remain in the bath of iced water for 40 minutes to ensure that
drying.
solution is complete. Adjust the weight of the solution to
Identification 300 g and centrifuge the solution to expel any trapped air.
Determine the viscosity at 20° by Method C (2.4.28), using a
A. With constant stirring add a quantity containing 1.0 g of shear rate of 10 s-1.
the dried substance into 50 ml of carbon dioxide-free water
previously heated to 90°. Allow to cool, dilute to 100 ml with
carbon dioxide-free water and continue stirring until solution Loss on drying (2.4.19). Not more than 5.0 per cent, determined
is complete (solution A). Heat 10 ml of solution A on a water- on 1.0 g by drying in an oven at 105°.
bath with stirring. At temperatures above 40° the solution Assay. Weigh accurately about 50 mg in a hard gelatin capsule
becomes cloudy or a flocculent precipitate is formed. On shell place the capsule and the contents in a 50-ml boiling
cooling, the solution becomes clear. flask and carry out the determination of methoxyl (2.3.29).
B. To 10 ml of solution A add 0.3 ml of 2 M acetic acid and 2.5 1ml of 0.1 M sodium thiosulphate is equivalent to 0.0005172 g
ml of a 10.0 per cent w/v solution of tannic acid; a yellowish of methoxyl (-OCH3) groups.
749
METHYLDOPA IP 2007
Storage. Store protected from light and moisture. D. To 10 mg add 3 drops of a 0.4 per cent w/v solution of
Labelling. The label states the apparent viscosity in millipascal ninhydrin in sulphuric acid; a dark purple colour is produced
seconds of a 2 per cent w/w solution. within 5 to 10 minutes. Add 0.15 ml of water; the colour changes
to pale brownish yellow.
Tests
Methyldopa Appearance of solution. A 4.0 per cent w/v solution in 1 M
hydrochloric acid is not more intensely coloured than
COOH reference solution BYS6 or BS6 (2.4.1).
H2N CH3 , 11 /2H2O Acidity. Dissolve 1.0 g in 100 ml of carbon dioxide-free water
HO with the aid of heat, add 0.15 ml of methyl red solution and
OH titrate with 0.1 M sodium hydroxide; not more than 0.5 ml is
required to produce a pure yellow colour.
C10H13NO4, 1½ H2O Mol. Wt. 238.2 Optical rotation (2.4.22). –1.10° to –1.23°, determined in a
Methyldopa is 3-(3,4-dihydroxyphenyl)-2-methyl-L-alanine solution prepared by dissolving a quantity containing 2.2 g of
sesquihydrate. the anhydrous substance in 50.0 ml of aluminium chloride
solution.
Methyldopa contains not less than 98.5 per cent and not more
than 101.0 per cent of C10H13NO4, calculated on the anhydrous 3-Methoxy compound and related substances. Determine by
basis. thin-layer chromatography (2.4.17), coating the plate with
microcrystalline cellulose.
Description. A white to yellowish white, fine powder which
may contain friable lumps. Mobile phase. A mixture of 65 volumes of 1-butanol, 25
volumes of water and 15 volumes of glacial acetic acid.
Identification Test solution. Dissolve 0.1 g of the substance under
Test A may be omitted if tests B, C and D are carried out. Tests examination in 10 ml of a mixture of 96 volumes of methanol
B, C and D may be omitted if test A is carried out. and 4 volumes of 7 M hydrochloric acid.
A. Determine by infrared absorption spectrophotometry (2.4.6). Reference solution (a). A 0.005 per cent w/v solution of
Compare the spectrum with that obtained with methyldopa 3-methoxymethyldopa RS in methanol.
RS or with the reference spectrum of methyldopa. Reference solution (b). A mixture of equal volumes of the test
B. When examined in the range 230 nm to 360 nm (2.4.7), a solution and reference solution (a).
0.004 per cent w/v solution in 0.1 M hydrochloric acid shows Apply to the plate 10 µl of each of the test solution and reference
an absorption maximum only at about 280 nm; absorbance at solution (a) and 20 µl of reference solution (b). After
about 280 nm, about 0.46. development, dry the plate immediately in a current of warm
C. Determine by thin-layer chromatography (2.4.17), coating air and spray with a mixture of 5 volumes of a 5 per cent w/v
the plate with microcrystalline cellulose. solution of sodium nitrite and 45 volumes of a 0.3 per cent
w/v solution of 4-nitroaniline in a mixture of 80 volumes of
Mobile phase. A mixture of 50 volumes of 1-butanol, 25 hydrochloric acid and 20 volumes of water. Dry it in a current
volumes of glacial acetic acid and 25 volumes of water. of warm air and spray with a 20 per cent w/v solution of sodium
Test solution. Dissolve 0.1 g of the substance under carbonate and examine immediately. Any secondary spot in
examination in 10 ml of 1 M hydrochloric acid. the chromatogram obtained with the test solution is not more
Reference solution. A 1 per cent w/v solution of methyldopa reference solution (a). The test is not valid unless the
RS in 1 M hydrochloric acid. chromatogram obtained with reference solution (b) shows two
Apply to the plate 5 µl of each solution. After development, clearly separated spots.
dry the plate in a current of warm air, and spray with a solution Heavy metals (2.3.13). Dissolve 2.0 g in 10 ml of water, add 2 ml
freshly prepared by mixing equal volumes of a 10 per cent w/ of dilute acetic acid and dilute to 25 ml with water. The solution
v solution of ferric chloride and a 5 per cent w/v solution of complies with the limit test for heavy metals, Method B
potassium ferricyanide. The principal spot in the (10 ppm).
that in the chromatogram obtained with the reference solution. Sulphated ash (2.3.18). Not more than 0.1 per cent.
750
IP 2007 METHYLERGOMETRINE MALEATE
Water (2.3.43). 10.0 to 13.0 per cent, determined on 0.4 g. Tests

Assay. Weigh accurately about 0.4 g and dissolve in 15 ml of Optical rotation (2.4.22). – 0.98° to –1.09°, determined on a
anhydrous formic acid, 30 ml of anhydrous glacial acetic solution prepared by dissolving an accurately weighed
acid and 30 ml of dioxan. Titrate with 0.1 M perchloric acid, quantity of the residue obtained in the Identification test
using crystal violet solution as indicator. Carry out a blank containing 0.39 g of C10H13NO4 in sufficient aluminium
titration. chloride solution to produce 10.0 ml. The content of C10H13NO4
1 ml of 0.1 M perchloric acid is equivalent to 0.02112 g of in the residue used for the test may be determined by titrating
C10H13NO4. with 0.1 M perchloric acid, using 0.2 g of the residue, crystal
violet solution as indicator.
C10H13NO4.
Methyldopa Tablets Assay. Weigh and powder 20 tablets. Weigh accurately a
Methyldopa Tablets contain not less than 95.0 per cent and quantity of the powder containing about 0.1 g of anhydrous
not more than 105.0 per cent of the stated amount of methyldopa, dissolve as completely as possible in sufficient
anhydrous methyldopa, C10H13NO4. The tablets are coated. 0.05 M sulphuric acid to produce 100.0 ml and filter. To 5.0 ml
of the filtrate add 2 ml of ferrous sulphate-citrate solution, 8
Identification ml of glycine buffer solution and sufficient water to produce
Remove the coating from a suitable quantity of the tablets by 100.0 ml. Measure the absorbance of the resulting solution at
washing with chloroform. To a quantity of the powdered tablet the maximum at about 545 nm (2.4.7). Calculate the content of
cores containing 5 g of anhydrous methyldopa add 35 ml of a C10H13NO4 taking 89 as the specific absorbance at 545 nm.
mixture of equal volumes of chloroform and methanol and Storage. Store protected from light and moisture.
shake for 3 minutes. Centrifuge and discard the supernatant
liquid. Repeat the operation with a further 35 ml of a mixture of
equivalent amount of anhydrous methyldopa.
equal volumes of chloroform and methanol. Dry the residue
in a current of nitrogen, add 20 ml of methanol and 15 ml of
2 M hydrochloric acid, shake for 2 minutes and filter. Adjust
the pH of the filtrate to 4.9 with 5 M ammonia, allow to stand Methylergometrine Maleate
for several hours at 2° to 8° and filter. Wash the precipitate Methylergonovine Maleate
with 15 ml of water and dry it at 50° at a pressure not exceeding
0.7 kPa for 3 hours. Reserve a portion of the residue for the
H
test for Specific optical rotation. The remainder of the residue O N
complies with tests A and B. CH3
A. Determine by infrared absorption spectrophotometry (2.4.6). OH COOH
Compare the spectrum with that obtained with methyldopa ,
N
RS or with the reference spectrum of methyldopa. CH3
H COOH
an absorption maximum only at about 280 nm; absorbance at HN
about 280 nm, about 0.46.
C20H25N3O2,C4H4O4 Mol. Wt. 455.5
C. To a quantity of the powdered tablets containing 10 mg of
Methylergometrine Maleate is 9,10-didehydro-N-[(S)-1-
anhydrous methyldopa, add 3 drops of a 0.4 per cent w/v
(hydroxymethyl)propyl]-6-methylergoline-8β-carboxamide
solution of ninhydrin in sulphuric acid; a dark purple colour
hydrogen maleate.
is produced within 5 to 10 minutes. Add 0.15 ml of water; the
colour changes to pale brownish yellow. Methylergometrine Maleate contains not less than 95.0 per
cent and not more than 105.0 per cent of C20H25N3O2,C4H4O4,
D. To 10 mg of the powdered tablets add 2 ml of 0.1 M sulphuric
acid, 2 ml of ferrous sulphate-citrate solution and 0.5 ml of
dilute ammonia solution; a dark purple colour is immediately Description. A white or faintly yellow, crystalline powder;
produced. odourless.
751
METHYLERGOMETRINE INJECTION IP 2007
Identification benzaldehyde reagent, mix, cool in running water for 5 minutes

and add sufficient reagent to produce 10.0 ml. Measure the
A. Determine by infrared absorption spectrophotometry (2.4.6). absorbance of the resulting solution at the maximum at about
Compare the spectrum with that obtained with 550 nm (2.4.7), using as the blank a solution prepared in the
methylergometrine maleate RS. same manner omitting the substance under examination.
B. In the test for Related substances, the principal spot in the Calculate the content of C 20H25N3O 2, C 4H 4O4 from the
chromatogram obtained with the test solution corresponds to absorbance obtained by repeating the Assay using
that in the chromatogram obtained with reference solution (a). ergometrine maleate RS in place of the substance under
examination.
C. A 1 per cent w/v solution shows a blue fluorescence.
1 mg of ergometrine maleate RS is equivalent to 1.032 mg of
D. Dissolve 0.25 mg in 1 ml of glacial acetic acid containing
C20H25N3O2, C4H4O4.
a trace of ferric chloride solution and add carefully 1 ml of
sulphuric acid and shake well; a deep blue colour is produced. Storage. Store protected from light, in an atmosphere of
nitrogen, at a temperature between 2° to 8°.
Tests
Specific optical rotation (2.4.22). +44.0° to +50°, determined at Methylergometrine Injection
20° in a 0.5 per cent w/v solution. Methylergometrine Maleate Injection; Methylergonovine
Related substances. Protect the solutions from light Maleate Injection; Methylergonovine Injection
throughout the test. Methylergometrine Injection is a sterile solution of
Determine by thin-layer chromatography (2.4.17), coating the Methylergometrine Maleate in Water for Injections free from
plate with silica gel G. dissolved air.
Mobile phase. A mixture of 10 volumes of chloroform and Methylergometrine Injection contains not less than 90.0 per
1 volume of methanol. cent and not more than 110.0 per cent of the stated amount of
methylergometrine maleate, C20H25N3O2,C4H4O4.
examination in 10 ml of methanol. Identification
Reference solution (a). A 0.4 per cent w/v solution of A. In the test for Related substances, the principal spot in the
methylergometrine maleate RS in methanol. chromatogram obtained with the test solution corresponds to
Reference solution (b). A 0.012 per cent w/v solution of that in the chromatogram obtained with reference solution (a).
methylergometrine maleate RS in methanol. B. It exhibits a blue fluorescence.
Place a beaker containing 25 ml of strong ammonia solution C. To a volume containing 0.1 mg of Methylergometrine
in the developing chamber, cover the chamber and allow to Maleate add 0.5 ml of water and 2 ml of 4-dimethyl-
equilibrate for 30 minutes. Apply to the plate 25 µl of each aminobenzaldehyde solution; after a few minutes a deep blue
solution. After development, dry the plate in air and examine colour is produced.
in ultraviolet light at 254 nm. Spray the plate with a solution
containing 0.8 g of 4-dimethylaminobenzaldehyde in a mixture Tests
of 90 ml of ethanol and 10 ml of sulphuric acid. Dry in a
pH (2.4.24). 2.7 to 3.5.
current of warm air for about 2 minutes. Any secondary spot
in the chromatogram obtained with the test solution is not Related substances. Protect the solutions from light
more intense than the spot in the chromatogram obtained throughout the test.
with reference solution (b). Determine by thin-layer chromatography (2.4.17), coating the
Sulphated ash (2.3.18). Not more than 0.1 per cent. plate with silica gel G.
Loss on drying (2.4.19). Not more than 2.0 per cent, determined Mobile phase. A mixture of 10 volumes of chloroform and 1
on 1.0 g by drying at 80° at a pressure not exceeding 2.7 kPa volume of methanol.
for 3 hours. Test solution. Transfer a volume containing 1 mg of
Assay. Weigh accurately about 20 mg and dissolve in sufficient Methylergometrine Maleate to a separating funnel, add 1 ml
water to produce 100.0 ml; dilute 20.0 ml of this solution to of sodium bicarbonate solution and extract with three
100.0 ml with water. To 3.0 ml add 6.0 ml of dimethylamino- quantities, each of 5 ml, of chloroform. Evaporate the combined
752
IP 2007 METHYLERGOMETRINE TABLETS
extracts to dryness at room temperature at a pressure not B. Extract a quantity of the powdered tablets containing 1 mg
exceeding 0.7 kPa. Dissolve the residue in 0.25 ml of methanol of Methylergometrine Maleate with 10 ml of water, filter and
and centrifuge, if necessary. wash the residue with sufficient water to produce 10 ml; the
Reference solution (a). A 0.4 per cent w/v solution of solution has a blue fluorescence.
methylergometrine maleate RS in methanol. C. To 2 ml of the solution obtained in test B add 4 ml of 4-
Reference solution (b). A 0.012 per cent w/v solution of dimethylaminobenzaldehyde solution; a deep blue colour is
methylergometrine maleate RS in methanol. produced after a few minutes.
Place a beaker containing 25 ml of strong ammonia solution Tests
in the developing chamber, cover the chamber and allow to
equilibrate for 30 minutes. Apply to the plate 25 µl of each Related substances. Protect the solutions from light
solution. After development, dry the plate in air and examine throughout the test.
in ultraviolet light at 254 nm. Spray the plate with a solution Determine by thin-layer chromatography (2.4.17), coating the
containing 0.8 g of 4-dimethylaminobenzaldehyde in a mixture plate with silica gel G.
volume of methanol.
more intense than the spot in the chromatogram obtained Test solution. To a quantity of the powdered tablets containing
with reference solution (b). 1 mg of Methylergometrine Maleate add 5 ml of water, 1 ml of
Other tests. Complies with the tests stated under Parenteral sodium bicarbonate solution and 2 ml of chloroform. Shake,
Preparations (Injections). allow to separate and filter the chloroform layer through a
plug of cotton moistened with chloroform. Repeat the
Assay. Protect the solutions from light throughout the Assay. extraction with a further 2 ml of chloroform and filter. Evaporate
To 1.0 ml add sufficient water to produce a solution containing the combined extracts to dryness at room temperature at a
0.04 mg of Methylergometrine Maleate per ml. To 3.0 ml add pressure not exceeding 0.7 kPa, dissolve the residue in 0.25 ml
6.0 ml of dimethylaminobenzaldehyde reagent, mix, cool in of methanol and centrifuge, if necessary.
running water for 5 minutes and add sufficient reagent to Reference solution (a). A 0.4 per cent w/v solution of
produce 10.0 ml. Measure the absorbance of the resulting methylergometrine maleate RS in methanol.
blank a solution prepared in the same manner omitting the Reference solution (b). A 0.012 per cent w/v solution of
substance under examination. Calculate the content of methylergometrine maleate RS in methanol.
C 20 H 25 N 3O 2,C 4H 4 O 4 from the absorbance obtained by Place a beaker containing 25 ml of strong ammonia solution
repeating the Assay using ergometrine maleate RS in place in the developing chamber, cover the chamber and allow to
of the substance under examination. equilibrate for 30 minutes. Apply to the plate 25 µl of each
1 mg of ergometrine maleate RS is equivalent to 1.032 mg of solution. After development, dry the plate in air and examine
C20H25N3O2,C4H4O4. in ultraviolet light at 254 nm. Spray the plate with a solution
containing 0.8 g of 4-dimethylaminobenzaldehyde in a mixture
Methylergometrine Tablets more intense than the spot in the chromatogram obtained
Methylergometrine Maleate Tablets; Methylergonovine with reference solution (b).
Maleate Tablets; Methylergonovine Tablets Uniformity of content. Comply with the test stated under
Methylergometrine Tablets contain not less than 90.0 per cent Tablets.
and not more than 110.0 per cent of the stated amount of Crush one tablet and transfer to a separating funnel with the
methylergometrine maleate, C20H25N3O2,C4H4O4. The tablets aid of not more than 5 ml of water and add 3 ml of a 5 per cent
may be coated. w/v solution of sodium carbonate. Extract with four quantities,
each of 5 ml, of chloroform. Filter the extracts through a plug
Identification
of cotton moistened with chloroform into a 100-ml separating
A. In the test for Related substances, the principal spot in the funnel. Add 2.0 ml of water and 10.0 ml of 4-dimethylamino-
chromatogram obtained with the test solution corresponds to benzaldehyde solution.and shake vigorously for at least 90
that in the chromatogram obtained with reference solution (a). seconds. Allow to stand for 30 minutes and discard the
753
METHYLPARABEN IP 2007
chloroform layer. Transfer the aqueous layer to a stoppered- Description. Ccolourless crystals or white, crystalline powder.
tube and allow to stand for 60 minutes. Measure the absorbance
of the resulting solution at the maximum at about 550 nm (2.4.7), Identification
using as the blank a mixture of 2.0 ml of water and 10.0 ml of 4-
dimethylaminobenzaldehyde solution. Calculate the content
of C19H23N3O2,C4H4O4 in the tablet from the absorbance
obtained by carrying out the following operation A. Determine by infrared absorption spectrophotometry (2.4.6).
simultaneously. Weigh accurately about 12 mg of ergometrine Compare the spectrum with that obtained with methylparaben
maleate RS and dissolve in sufficient water to produce 200.0 RS.
ml. To 2.0 ml add 10.0 ml of 4-dimethylaminobenzaldehyde B. When examined in the range 230 nm to 360 nm (2.4.7), a
solution, mix and cool in running water for 5 minutes. Measure 0.0005 per cent w/v solution in ethanol (95 per cent) shows
the absorbance of the resulting solution at the maximum at an absorption maximum at about 258 nm; absorption at about
about 545 nm, using as the blank a mixture of 2.0 ml of water 258 nm, 0.52 to 0.56.
and 10.0 ml of 4-dimethylaminobenzaldehyde solution.
C. Boil 10 mg with 10 ml of water, cool and add 0.05 ml of ferric
1 mg of ergometrine maleate RS is equivalent to 1.032 mg of chloride solution; a reddish violet colour is produced.
C20H25N3O2,C4H4O4. D. Dissolve 0.1 g in 2 ml of ethanol (95 per cent), boil and add
Other tests. Comply with the tests stated under Tablets. 0.5 ml of mercuric nitrate solution; a precipitate is formed and
the supernatant liquid becomes red.
quantity of the powder containing about 2 mg of Tests
Methylergometrine Maleate, dissolve in 50 ml of a 1 per cent
w/v solution of tartaric acid. To 3.0 ml add 6.0 ml of Appearance of solution. A 10.0 per cent w/v solution in ethanol
dimethylaminobenzaldehyde reagent, mix, cool in running (95 per cent) is clear (2.4.1), and not more intensely coloured
water for 5 minutes and add sufficient reagent to produce 10.0 than reference solution BYS6 (2.4.1).
ml. Measure the absorbance of the resulting solution at the Acidity. Dissolve 1.0 g in sufficient ethanol (95 per cent) to
maximum at about 550 nm (2.4.7), using as the blank a solution produce 10 ml. To 2 ml of the solution add 3 ml of ethanol (95
prepared in the same manner omitting the substance under per cent), 5 ml of carbon dioxide-free water and 0.1 ml of
examination. Calculate the content of C20H25N3O2,C4H4O4 from bromocresol green solution. Not more than 0.1 ml of 0.1 M
the absorbance obtained by repeating the Assay using sodium hydroxide is required to change the colour of the
ergometrine maleate RS in place of the substance under solution.
examination.
1 mg of ergometrine maleate RS is equivalent to .001032 g of (2.4.17), coating the plate with silica gel HF254.
C20H25N3O2,C4H4O4.
Storage. Store protected from light and moisture. 10 volumes of ethyl acetate and 2 volumes of anhydrous
formic acid.
Methylparaben examination in 10 ml of methanol.
Methyl Hydroxybenzoate
COOCH3 Apply to the plate 5 µl of each solution. After development,
dry the plate in a current of hot air and examine in ultraviolet
OH Chlorides (2.3.12). Heat 2.0 g with 100 ml of water, cool, add
sufficient water to restore the original volume, and filter. 25 ml
C8H8O3 Mol. Wt. 152.2
of the filtrate complies with the limit test for chlorides
Methylparaben is methyl 4-hydroxybenzoate. (500 ppm).
Methylparaben contains not less than 99.0 per cent and not Sulphates. To 10 ml of the filtrate obtained in the test for
more than 101.0 per cent of C8H8O3. Chloride add 0.15 ml of dilute hydrochloric acid and 0.1 ml of
754
IP 2007 METHYLPREDNISOLONE
barium chloride solution; no turbidity is produced within 10 B. Determine by thin-layer chromatography (2.4.17), coating
minutes. the plate with silica gel G.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Solvent mixture. A mixture of 90 volumes of acetone and 10
Assay. Weigh accurately about 80 mg, transfer to a glass- volumes of formamide.
stoppered flask, add 25 ml of 2 M sodium hydroxide and boil Mobile phase. Chloroform.
gently under a reflux condenser for 30 minutes. Cool and add
25.0 ml of 0.0333 M potassium bromate, 5 ml of a 12.5 per cent Test solution. Dissolve 25 mg of the substance under
w/v solution of potassium bromide and 40 ml of glacial acetic examination in 10 ml of the solvent mixture.
acid, cool in ice, add 10 ml of hydrochloric acid, immediately Reference solution (a). Dissolve 25 mg of methylprednisolone
stopper the flask and allow to stand for 15 minutes. Add 15 ml RS in 10 ml of the solvent mixture.
of potassium iodide solution, mix and titrate the liberated
iodine with 0.1 M sodium thiosulphate using 2 ml of starch Reference solution (b). Mix equal volumes of the test solution
solution, added towards the end of the titration, as indicator. and reference solution (a).
Repeat the operation without the substance under examination. Place the dry plate in a tank containing a shallow layer of the
The difference between the titrations represents the amount solvent mixture, allow the solvent mixture to ascend to the
of potassium bromate required. The volume of 0.0333 M top, remove the plate from the tank and allow the solvent to
potassium bromate is equivalent to half of the volume of 0.1 evaporate. Use within 2 hours, with the flow of the mobile
M sodium thiosulphate required for the titration. phase in the direction in which the aforementioned treatment
1 ml of 0.0333 M potassium bromate is equivalent to 0.005072 was done.
g of C8H8O3. Apply to the plate 2 µl of each solution. Allow the mobile
Storage. Store protected from light and moisture. phase to rise 12 cm. Dry the plate in a current of warm air, allow
Methylprednisolone in daylight and in ultraviolet light at 365 nm. The principal
O
OH corresponds to that in the chromatogram obtained with
H3C
HO OH reference solution (a). The principal spot in the chromatogram
H3C H compact spot.
H H C. Dissolve about 2 mg in 2 ml of sulphuric acid by shaking
O and allow to stand for 5 minutes; an intense red colour is
CH3 produced and the solution exhibits a reddish brown
fluorescence when examined in ultraviolet light at 365 nm.
C22H30O5 Mol. Wt. 374.5 Add the solution to 10 ml of water and mix; the colour fades
and the solution exhibits a yellowish green fluorescence in
Methylprednisolone is 11β,17α,21-trihydroxy-6α-
ultraviolet light at 365 nm.
methylpregna-1,4-diene-3,20-dione.
Methylprednisolone contains not less than 96.0 per cent and Tests
dried basis. Specific optical rotation (2.4.22). +79.0° to +86.0°, determined
Identification solution in ethanol (95 per cent) at the maximum at about
243 nm, 0.38 to 0.40.
Test A may be omitted if tests B and C are carried out. Test C
may be omitted if tests A and B are carried out. Related substances. Determine by liquid chromatography
(2.4.14).
Compare the spectrum with that obtained with Test solution. Dissolve 25 mg of the substance under
methylprednisolone RS or with the reference spectrum of examination in a mixture of equal volumes of acetonitrile and
methylprednisolone. methanol and dilute to 10 ml with the same solvent mixture.
755
METHYLPREDNISOLONE TABLETS IP 2007
Reference solution (a). Dissolve 2.0 mg of methylprednisolone solution, the area of any peak other than the principal peak, is
RS and 2.0 mg of betamethasone RS in mobile phase A and not greater than half the area of the principal peak in the
dilute to 200 ml with the same mobile phase. chromatogram obtained with reference solution (b) (0.5 per
Reference solution (b). Dilute 1 ml of the test solution to cent); the sum of the areas of all the peaks other than the
100 ml with mobile phase A principal peak, is not greater than twice the area of the principal
Chromatographic system (b) (2 per cent). Ignore any peak due to the blank and any peak
– a stainless steel column 25 cm x 4.6 mm, packed with with an area less than 0.05 times the area of the principal peak
octadecylsilane chemically bonded to silica gel (5 µm), in the chromatogram obtained with reference solution (b).
– mobile phase A. 250 volumes of acetonitrile and
700 volumes of water mixed, allowed to equilibrate and
adjusted to 1000 volumes with water and mixed, Assay. Weigh accurately about 0.1 g and dissolve in sufficient
B. acetonitrile, ethanol to produce 100.0 ml and mix. Dilute 2.0 ml of this
– flow rate. 2.5 ml per minute, solution to 100.0 ml with ethanol and mix well. Determine the
– a linear gradient programme using the conditions given absorbance of the resulting solution (2.4.7) at the maximum at
below, about 243 nm. Calculate the content of C22H29FO5 taking 395
– spectrophotometer set at 254 nm, as the specific absorbance at 243 nm.
phase A phase B
0 100 0 Isocratic Methylprednisolone Tablets
15 100 100 begin
linear gradient Methylprednisolone Tablets contain not less than 90.0 per
40 0 100 end chromatogram, methylprednisolone, C22H30O5.
return to 100 A
41 100 0 begin equilibration Identification
with A
Extract a quantity of the powdered tablets containing 50 mg of
46=0 100 0 end equilibration,
Methylprednisolone with 100 ml of chloroform, filter and
begin next
evaporate the filtrate to dryness. The residue complies with
chromatogram
the following tests.
Equilibrate the column for at least 30 minutes with mobile
phase B and then with mobile phase A for 5 minutes. For A. Determine by infrared absorption spectrophotometry (2.4.6).
subsequent operations use the conditions described from Compare the spectrum with that obtained with
40 to 46 minutes. methylprednisolone RS or with the reference spectrum of
methylprednisolone.
principal peak in the chromatogram obtained with reference B. Determine by thin-layer chromatography (2.4.17), coating
solution (b) is at least 50 per cent of the full scale of the the plate with silica gel G.
recorder. Solvent mixture. A mixture of 90 volumes of acetone and 10
Inject reference solution (a). When the chromatograms are volumes of formamide.
recorded, the retention times are; methylprednisolone about Mobile phase. A mixture of 30 volumes of toluene and 10
11.5 minutes, and betamethasone about 12.5 minutes. The volumes of chloroform.
test is not valid unless the resolution between the peaks
corresponding to methylprednisolone and betamethasone is Test solution. Dissolve 25 mg of the residue in 10 ml of the
at least 1.5; if necessary, adjust the concentration of acetonitrile solvent mixture.
in mobile phase A. Reference solution (a). Dissolve 25 mg of methylprednisolone
Inject separately a mixture of equal volumes of acetonitrile acetate RS in 10 ml of the solvent mixture.
and methanol as a blank, the test solution and reference Reference solution (b). Mix equal volumes of the test solution
solution (b). In the chromatogram obtained with the test and reference solution (a).
756
IP 2007 METHYLPREDNISOLONE TABLETS
Place the dry plate in a tank containing a shallow layer of the Dissolution (2.5.2).
solvent mixture, allow the solvent mixture to ascend to the Apparatus. No 2
was done. Withdraw a suitable volume of the medium and filter. Measure
Apply to the plate 2 µl of each solution. Allow the mobile the absorbance of a layer of suitable thickness of the filtered
phase to rise 12 cm. Dry the plate in a current of warm air, allow solution at the maximum at about 246 nm (2.4.7). Calculate the
the solvent to evaporate, heat at 120° for 15 minutes and spray content of C22H30O5 in the medium taking 400 as the specific
the hot plate with ethanolic sulphuric acid (20 per cent v/v). absorbance at 246 nm.
Heat at 120° for a further 10 minutes, allow to cool and examine D. Not less than 70 per cent of the stated amount of C22H30O5.
Tablets.
reference solution (a). The principal spot in the chromatogram To one tablet add 0.5 ml of water (in the case of tablets
obtained with reference solution (b) appears as a single, containing 10 mg or less) or 1.0 ml of water (in the case of
compact spot. tablets containing more than 10 mg). Allow the tablet to stand
for about 2 minutes, then swirl to disperse the tablet. Add
Tests 5.0 ml of the internal standard used in the assay for each mg of
Related substances. Determine by liquid chromatography methylprednisolone, shake for 15 minutes, filter and centrifuge.
(2.4.14). Use the filtrate as the test solution.
Diluting solution. A filtered mixture of 72 volumes of water, Determine by liquid chromatography (2.4.14) using the
25 volumes of tetrahydrofuran and 3 volumes of glacial acetic chromatographic system and the reference solution described
acid. under Assay.
Test solution. Extract a quantity of the powdered tablets Calculate the content of C22H30O5 in the tablet.
containing 25 mg of Methylprednisolone with the diluting Other tests. Comply with the tests stated under Tablets.
solution and dilute to 25 ml with the same solvent. Filter and
centrifuge if necessary. Assay. Determine by liquid chromatography (2.4.14).
Reference solution. A 0.001 per cent w/v solution of Internal standard solution. Weigh accurately a suitable
methylprednisolone RS in the diluting solution. quantity of prednisolone in a 3 per cent v/v solution of glacial
acetic acid in chloroform to obtain a solution having a known
concentration of about 0.2 mg per ml of prednisolone.
octadecylsilyl silica gel (3 to 10 µm), Test solution. Weigh and powder 20 tablets. Weigh accurately
– mobile phase: a mixture of 149 volumes of water, 40 a quantity of the powder containing about 10 mg of
volumes of tetrahydrofuran, 10 volumes of Methylprednisolone transfer to a suitable container and add
dimethysulfoxide and 1 volume of butanol, 2.5 ml of water. Swirl to form a slurry. Add 50.0 ml of the
– flow rate. 1 ml per minute, internal standard solution, and shake for 15 minutes. Filter
– spectrophotometer set at 254 nm, and centrifuge a portion of the filtrate if necessary and use
– a 10 µl loop injector. this as the test solution.
Inject the reference solution. The column efficiency is not Reference solution. Weigh accurately a suitable quantity of
less than 800 theoretical plates and the relative standard methylprednisolone RS in the internal standard solution to
deviation for replicate injections is not more than 5.0 per cent. obtain a solution having a known concentration of about
0.2 mg per ml of methylprednisolone.
Inject the test solution and the reference solution. In the
chromatogram obtained with the test solution, the area of any Chromatographic system
peak other than the principal peak is not greater than the area – stainless steel column 25 cm x 4 mm, packed with porous
of the principal peak in the chromatogram obtained with the silica particles (3 to 10 µm),
reference solution (1.0 per cent). The sum of the areas of all – mobile phase: a mixture of 475 volumes of butyl chloride,
the peaks other than the principal peak is not greater than 475 volumes of water-saturated butyl chloride, 70
twice the area of the principal peak in the chromatogram volumes of tetrahydrofuran, 35 volumes of
obtained with the reference solution (2.0 per cent). methanol,and 30 volumes of glacial acetic acid,
757
METHYLPREDNISOLONE ACETATE IP 2007
– flow rate 1 ml per minute, Test solution. Dissolve 25 mg of the substance under
– spectrophotometer set at 254 nm, examination in 10 ml of the solvent mixture.
– a 10 µl loop injector. Reference solution (a). Dissolve 25 mg of methylprednisolone
Inject the reference solution. The resolution between acetate RS in 10 ml of the solvent mixture.
methylprednisolone and prednisolone is not less than 4.0 and
the relative standard deviation for replicate injections is not
The relative retention times are 0.7 for prednisolone and
1.0 for methylprednisolone.
Calculate the content of C22H30O5 in the tablets. phase in the direction in which the aforementioned treatment
Storage. Store protected from light and moisture. was done.
Methylprednisolone Acetate the hot plate with ethanolic sulphuric acid (20 per cent v/v).
O in daylight and in ultraviolet light at 365 nm. The principal
O CH3 corresponds to that in the chromatogram obtained with
H3C O
HO OH
H3C H compact spot.
H H
Tests
O
CH3 Specific optical rotation (2.4.22). +97.0° to +105°, determined
C24H32O6 Mol. Wt. 416.5 Light absorption (2.4.7). Absorbance of a 0.001 per cent w/v
solution in ethanol at the maximum at about 240 nm, 0.34 to
Methylprednisolone Acetate is 11β,17α-dihydroxy-6α- 0.37. The ratio of the absorbance at the maximum at about 240
methyl-3, 20-dioxopregna-1,4-dien-21-yl acetate. nm to that at about 263 nm is 1.50 to 1.70.
Methylprednisolone Acetate contains not less than 96.0 per Related substances. Determine by liquid chromatography
cent and not more than 104.0 per cent of C24H32O6, calculated (2.4.14).
on the dried basis.
Description. A white or almost white, crystalline powder; examination in 5 ml of tetrahydrofuran and dilute to 10 ml with
odourless or almost odourless. water.
Identification Reference solution (a). Dissolve 4.0 mg of methylprednisolone
acetate RS and 4.0 mg of dexamethasone acetate RS in the
A. Determine by infrared absorption spectrophotometry (2.4.6). mobile phase and dilute to 20 ml with the mobile phase.
methylprednisolone acetate RS or with the reference spectrum Reference solution (b). Dilute 1 ml of the test solution to 50 ml
of methylprednisolone acetate. with the mobile phase.
B. Determine by thin-layer chromatography (2.4.17), coating Chromatographic system
the plate with silica gel G. – a stainless steel column 25 cm x 4.6 mm, packed with
(5 µm),
volumes of formamide.
– mobile phase: a mixture of 260 ml of tetrahydrofuran
Mobile phase. A mixture of 30 volumes of toluene and 10 and 760 ml of water, allowed to equilibrate, diluted to
volumes of chloroform. 1000 ml with water and mixed,
758
IP 2007 METHYLPREDNISOLONE ACETATE INJECTION
– flow rate. 1 ml per minute, Identification

– a 20 µl loop injector. Dilute a volume containing 0.1 g of Methylprednisolone
Acetate to 5 ml with water, centrifuge and discard the
Equilibrate the column with the mobile phase for about 45 supernatant liquid. Wash the residue with five quantities, each
minutes. of 5 ml, of water, resuspending the residue in water each time.
Adjust the sensitivity of the system so that the height of the Centrifuge and discard the washings. The residue, after drying
principal peak in the chromatogram obtained with reference at 105° for 3 hours, complies with the following tests.
solution (b) is at least 50 per cent of the full scale of the A. Determine by infrared absorption spectrophotometry (2.4.6).
recorder. Compare the spectrum with that obtained with
Inject reference solution (a). The retention times are: methylprednisolone acetate RS or with the reference spectrum
methylprednisolone acetate, about 43 minutes and of methylprednisolone acetate.
dexamethasone acetate about 57 minutes. The test is not valid B. Determine by thin-layer chromatography (2.4.17), coating
unless the resolution between the peaks corresponding to the plate with silica gel G.
methylprednisolone acetate and dexamethasone acetate is not
less than 6.5. If necessary, adjust the concentration of water
volumes of formamide.
in the mobile phase.
Inject the test solution and reference solution (b). Continue volumes of chloroform.
the chromatography for 1.5 times the retention time of the
principal peak. In the chromatogram obtained with the test Test solution. Dissolve 25 mg of the residue in 10 ml of the
solution, the sum of the areas of all the peaks other than the solvent mixture.
principal peak is not greater than the area of the principal peak Reference solution (a). Dissolve 25 mg of methylprednisolone
in the chromatogram obtained with reference solution (b) acetate RS in 10 ml of the solvent mixture.
(1.0 per cent). Ignore any peak due to the solvent and any Reference solution (b). Mix equal volumes of the test solution
peak with an area less than 0.025 times the area of the principal and reference solution (a).
(b). Place the dry plate in a tank containing a shallow layer of the
Sulphated ash (2.3.18). Not more than 0.2 per cent. top, remove the plate from the tank and allow the solvent to
Loss on drying (2.4.19). Not more than 1.0 per cent, determined evaporate. Use within 2 hours, with the flow of the mobile
on 1.0 g by drying in an oven at 105° for 3 hours. phase in the direction in which the aforementioned treatment
was done.
ethanol to produce 100.0 ml and mix. Dilute 1.0 ml of this Apply to the plate 2 µl of each solution. Allow the mobile
solution to 100.0 ml with ethanol and mix well. Determine the phase to rise 12 cm. Dry the plate in a current of warm air, allow
absorbance of the resulting solution (2.4.7) at the maximum at the solvent to evaporate, heat at 120° for 15 minutes and spray
about 243 nm. Calculate the content of C24H32O6, taking 355 as the hot plate with ethanolic sulphuric acid (20 per cent v/v).
the specific absorbance at 243 nm. Heat at 120° for a further 10 minutes, allow to cool and examine
Methylprednisolone Acetate Injection obtained with reference solution (b) appears as a single,
compact spot.
Methylprednisolone Acetate Injection is a sterile suspension
of Methylprednisolone Acetate in Water for Injections. Tests
Methylprednisolone Acetate Injection contains not less than pH (2.4.24). 3.5 to 7.0.
amount of methylprednisolone acetate, C24H32O6. Other tests. Complies with the tests stated under Parenteral
Description. A white suspension which settles on standing
but readily disperses on shaking. On examination under a Assay. Determine by liquid chromatography (2.4.14).
microscope, the particles are seen to be crystalline and rarely Mix 0.12 g of prednisone RS (internal standard) with 0.6 ml of
exceed 20 µm in diameter. glacial acetic acid, slowly add chloroform with the aid of
759
METOCLOPRAMIDE HYDROCHLORIDE IP 2007
ultrasound, shake to dissolve and dilute with sufficient Metoclopramide Hydrochloride contains not less than 99.0
chloroform to produce 20 ml (solution A). per cent and not more than 101.0 per cent of C14H22ClN3O2,HCl,
Test solution. Add 10 ml of solution A to an accurately calculated on the anhydrous basis.
measured quantity of the injection containing about 40 mg of Description. White or almost white crystals or crystalline
Methylprednisolone Acetate, add sufficient chloroform to powder.
produce 25.0 ml and shake for 5 minutes or until the aqueous
layer is clear; to 4.0 ml of the chloroform layer, add 30 ml of Identification
chloroform and 0.4 g of anhydrous sodium sulphate, shake Test A may be omitted if tests B, C and D are carried out. Tests
for 5 minutes, and use the clear solution. B and D may be omitted if tests A and C are carried out.
Reference solution. Dissolve 20.0 mg of methylprednisolone A. Determine by infrared absorption spectrophotometry (2.4.6).
acetate RS in 5 ml of solution A and add sufficient chloroform Compare the spectrum with that obtained with metoclopramide
to produce 100.0 ml. hydrochloride RS or with the reference spectrum of
Chromatographic system metoclopramide hydrochloride.
– a stainless steel column 25 cm x 4 mm, packed with B. Examine the chromatograms obtained in the test for Related
octadecylsilane chemically bonded to porous silica substances in ultraviolet light before spraying with the
(5 to 10 µm), 4-dimethylaminobenzaldehyde reagent. The principal spot
– mobile phase: a mixture of 30 volumes of glacial acetic in the chromatogram obtained with test solution (b)
acid and 35 volumes of methanol, 75 volumes of corresponds to that in the chromatogram obtained with
tetrahydrofuran, 475 volumes of water-saturated 1- reference solution (b).
chloro-butane and 475 volumes of
C. A 5 per cent w/v solution in carbon dioxide-free water
1-chloro-butane,
gives reaction A of chlorides (2.3.1).
– spectrophotometer set at 254 nm, D. Dissolve about 2 mg in 2 ml of water. The solution gives
– a 20 µl loop injector. the reaction of primary aromatic amines (2.3.1).
The assay is not valid unless the resolution between the peaks Tests
due to methylprednisolone and the internal standard is at
least 2.5.
Calculate the content of C22H29FO5 in the injection.
Storage. Store protected from light at a temperature not Related substances. Determine by thin-layer chromatography
exceeding 30°. The injection should not be allowed to freeze. (2.4.17), coating the plate with silica gel HF254.
Labelling. The label states (1) that the preparation is not to be Mobile phase. A mixture of 90 volumes of dichloromethane,
given by intravenous injection; (2) that the container should 14 volumes of methanol, 10 volumes of dioxan and 2 volumes
be shaken gently before a dose is withdrawn. of strong ammonia solution.
Metoclopramide Hydrochloride Test solution (b). Dissolve 0.4 g of the substance under
H
O N Reference solution (a). A 0.02 per cent w/v solution of the
N CH3 substance under examination in methanol.
OCH3 , HCl, H2O
CH3 Reference solution (b). A 0.4 per cent w/v solution of
metoclopramide hydrochloride RS in methanol.
Cl Reference solution (c). A 0.02 per cent w/v solution of N,N-
NH2 diethyl-ethylenediamine in methanol.
C14H22ClN3O2,HCl,H2O Mol. Wt. 354.3
Metoclopramide Hydrochloride is 4-amino-5-chloro-N-(2- light at 254 nm. Any secondary spot in the chromatogram
diethylaminoethyl)-2- methoxybenzamide hydrochloride obtained with test solution (a) is not more intense than the
monohydrate. spot in the chromatogram obtained with reference solution
760
IP 2007 METOCLOPRAMIDE SYRUP
(a). Spray the plate with dimethylaminobenzaldehyde reagent Related substances. Determine by thin-layer chromatography
and allow it to dry in air. Any secondary spot in the (2.4.17), coating the plate with silica gel HF254.
chromatogram obtained with test solution (a) that has not Mobile phase. A mixture of 90 volumes of dichloromethane,
been visualised in ultraviolet light at 254 nm is not more intense 14 volumes of methanol, 10 volumes of dioxan and 2 volumes
than the spot in the chromatogram obtained with reference of strong ammonia solution.
solution (c).
Test solution. Dilute a volume of the injection with methanol,
Heavy metals (2.3.13). 12 ml of a 10 per cent w/v solution in if necessary, to contain the equivalent of 0.5 per cent w/v of
carbon dioxide-free water complies with the limit test for anhydrous metoclopramide hydrochloride.
heavy metals, Method D ( 20 ppm). Use lead standard solution
(2 ppm Pb) to prepare the standard. Reference solution (a). Dilute 1 volume of the test solution to
200 volumes with methanol.
Reference solution (b). A 0.0025 per cent w/v solution of N,N-
Water (2.3.43). 4.5 to 5.5 per cent, determined on 0.5 g. diethylethylenediamine in methanol.
Assay. Weigh accurately about 0.25 g, dissolve in a mixture of Apply to the plate 10 µl of each solution. Allow the mobile
50 ml of ethanol (95 per cent) and 5.0 ml of 0.01 M phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
hydrochloric acid. Titrate with 0.1 M sodium hydroxide, light at 254 nm. Any secondary spot in the chromatogram
determining the end-point potentiometrically (2.4.25). Note obtained with the test solution is not more intense than the
the volume added between the two inflections. spot in the chromatogram obtained with reference solution
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03363 g of (a). Spray the plate with dimethylaminobenzaldehyde reagent
C14H22ClN3O2,HCl. and allow it to dry in air. Any secondary spot in the
Storage. Store protected from light and moisture. chromatogram obtained with the test solution that has not
been visualised in ultraviolet light at 254 nm is not more intense
solution (b).
Metoclopramide Injection
Metoclopramide Hydrochloride Injection Preparations (Injections).
Metoclopramide Injection is a sterile solution of Assay. Dilute an accurately measured volume containing about
Metoclopramide Hydrochloride in Water for Injections free 10 mg of anhydrous metoclopramide hydrochloride to 100.0
from dissolved air. It contains suitable buffering and stabilising ml with water. To 20.0 ml of this solution add 15 ml of 1.25 M
agents. sodium hydroxide and extract with three quantities, each of
Metoclopramide Injection contains not less than 90.0 per cent 30 ml, of chloroform, dry each extract with anhydrous sodium
and not more than 110.0 per cent of the stated amount of sulphate and filter. Dilute the combined extracts to 100.0 ml
anhydrous metoclopramide hydrochloride, C14H22ClN3O2,HCl. with chloroform and mix. Measure the absorbance of the
Description. A clear, colourless solution.
Calculate the content of C14H22ClN3O2,HCl, taking 265 as the
Identification specific absorbance at 305 nm.
A. Dilute a volume containing 10 mg of anhydrous Storage. Store protected from light.
metoclopramide hydrochloride to 500 ml with 0.01 M Labelling. The label states the strength in terms of the
hydrochloric acid. equivalent amount of anhydrous metoclopramide
When examined in the range 230 nm to 360 nm (2.4.7), the hydrochloride in a suitable dose-volume.
resulting solution shows absorption maxima at about 273 nm
and 309 nm.
B. To a volume containing 50 mg of anhydrous metoclopramide Metoclopramide Syrup
hydrochloride add 5 ml of water and 5 ml of a 1 per cent w/v
Metoclopramide Hydrochloride Syrup
solution of 4-dimethylaminobenzaldehyde in 1 M
hydrochloric acid; a yellow-orange colour is produced. Metoclopramide Syrup contains Metoclopramide
Hydrochloride in a suitable flavoured vehicle.
Metoclopramide Syrup contains not less than 90.0 per cent
Tests and not more than 110.0 per cent of the stated amount of
pH (2.4.24). 3.0 to 5.0. anhydrous metoclopramide hydrochloride, C14H22ClN3O2,HCl.
761
METOCLOPRAMIDE TABLETS IP 2007
Identification frequent shaking. Cool, dilute to 100 ml with 0.01 M

hydrochloric acid, filter and dilute 10 ml of the filtrate to 50 ml
To 50 ml add 5 M sodium hydroxide till the solution becomes with the same solvent.
alkaline and extract with three quantities, each of 40 ml, of
chloroform, dry each extract with anhydrous sodium sulphate. When examined in the range 230 nm to 360 nm (2.4.7), the
Evaporate the combined extracts to dryness on a water-bath. resulting solution shows absorption maxima at about 273 nm
The residue complies with the following tests. and 309 nm.
A. Dissolve 10 mg of the residue in 0.01 M hydrochloric acid B. Shake a quantity of the powdered tablets containing 50 mg
and dilute to 500 ml with 0.01 M hydrochloric acid. of anhydrous metoclopramide hydrochloride with 5 ml of water,
filter and add to the filtrate 5 ml of a 1 per cent w/v solution of
When examined in the range 230 nm to 360 nm (2.4.7), the 4-dimethylaminobenzaldehyde in 1 M hydrochloric acid; a
resulting solution shows absorption maxima at about 273 nm yellow-orange colour is produced.
and 309 nm.
B. To 25 mg of the residue add 2.5 ml of water and 2.5 ml of a Tests
1 per cent w/v solution of 4-dimethylaminobenzaldehyde in Related substances. Determine by thin-layer chromatography
1 M hydrochloric acid; a yellow-orange colour is produced. (2.4.17), coating the plate with silica gel HF254.
D. Dissolve about 2 mg in 2 ml of water. The solution gives Mobile phase. A mixture of 90 volumes of dichloromethane,
the reaction of primary aromatic amines (2.3.1). 14 volumes of methanol, 10 volumes of dioxan and 2 volumes
Tests
pH (2.4.24). 2.0 to 4.0. containing 0.1 g of anhydrous metoclopramide hydrochloride
Other tests. Complies with the tests stated under Oral Liquids. with 20 ml of methanol for 5 minutes and filter.
Assay. Dilute an accurately measured volume containing about Reference solution (a). Dilute 1 volume of the test solution to
10 mg of anhydrous metoclopramide hydrochloride to 100.0 200 volumes with methanol.
ml with water. To 20.0 ml of this solution add 15 ml of 1.25 M Reference solution (b). A 0.0025 per cent w/v solution of N,N-
sodium hydroxide and extract with three quantities, each of diethylethylenediamine in methanol.
30 ml, of chloroform, dry each extract with anhydrous sodium
sulphate and filter. Dilute the combined extracts to 100.0 ml Apply to the plate 10 µl of each solution. Allow the mobile
with chloroform and mix. Measure the absorbance of the phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
resulting solution at the maximum at about 305 nm (2.4.7). light at 254 nm. Any secondary spot in the chromatogram
Calculate the content of C14H22ClN3O2,HCl, taking 265 as the obtained with the test solution is not more intense than the
specific absorbance at 305 nm. spot in the chromatogram obtained with reference solution
(a). Spray the plate with dimethylaminobenzaldehyde reagent
Storage. Store protected from light and moisture. and allow it to dry in air. Any secondary spot in the
Labelling. The label states the strength in terms of the chromatogram obtained with the test solution that has not
equivalent amount of anhydrous metoclopramide been visualised in ultraviolet light at 254 nm is not more intense
hydrochloride in a suitable dose-volume. than the spot in the chromatogram obtained with reference
solution (b).
Tablets.
Metoclopramide Tablets
Powder one tablet and carry out the Assay beginning at the
Metoclopramide Hydrochloride Tablets words “add 50 ml of 0.1 M hydrochloric acid,....”.
Metoclopramide Tablets contain not less than 90.0 per cent Other tests. Comply with the tests stated under Tablets.
and not more than 110.0 per cent of the stated amount of Assay. Weigh and powder 20 tablets. Weigh accurately a
anhydrous metoclopramide hydrochloride, C14H22ClN3O2,HCl. quantity of the powder containing 10 mg of anhydrous
metoclopramide hydrochloride, add 50 ml of 0.1 M
Identification
hydrochloric acid, heat on a water-bath at 70° for 15 minutes,
A. Shake a quantity of the powdered tablets containing 10 mg cool, dilute to 100.0 ml with water and filter. To 20.0 ml of this
of anhydrous metoclopramide hydrochloride with 50 ml of solution add 15 ml of 1.25 M sodium hydroxide and extract
0.01 M hydrochloric acid and heat at 70° for 15 minutes with with three quantities, each of 30 ml, of chloroform, dry each
762
IP 2007 METOPROLOL TABLETS
extract with anhydrous sodium sulphate and filter. Dilute the Mobile phase. Chloroform. Pour 200 ml of chloroform in the
combined extracts to 100.0 ml with chloroform and mix. developing chamber containing several beakers, each
Measure the absorbance of the resulting solution at the containing 45 ml of strong ammonia solution and saturate for
maximum at about 305 nm (2.4.7). Calculate the content of 1½ hours by lining the walls with absorbent paper.
C14H22ClN3O2,HCl, taking 265 as the specific absorbance at Test solution. Dissolve 0.2 g of the substance under
305 nm. examination in 10 ml of chloroform.
Storage. Store protected from light and moisture. Reference solution (a). A 0.01 per cent w/v solution of the
Labelling. The label states the strength in terms of the substance under examination in chloroform.
equivalent amount of anhydrous metoclopramide Reference solution (b). A 0.004 per cent w/v solution of the
hydrochloride. substance under examination in chloroform.
dry the plate in air. Place the plate in a chamber of chlorine gas
Metoprolol Tartrate prepared by adding 5 ml of 5 M hydrochloric acid to a beaker
containing 0.5 g of potassium permanganate, and let the plate
OH H OH remain in the chamber for about a minute. Remove the plate
H
O N CH3 COOH from the chamber, allow to stand for a few minutes and spray
, HOOC
CH3 H OH it with a 0.5 per cent w/v solution of potassium iodide in
H3CO 2 starch solution. Any secondary spot in the chromatogram
(C15H25NO3)2,C4H6O6 Mol. Wt. 684.8 spot in the chromatogram obtained with reference solution (a)
Metoprolol Tartrate is (RS)-1-isopropylamino-3-p- (2- and not more than one such spot is more intense than the
methoxyethyl)phenoxypropan-2-ol (2R,3R)-tartrate. spot in the chromatogram obtained with reference solution
(b).
Metoprolol Tartrate contains not less than 99.0 per cent and
not more than 101.0 per cent of (C 15H25NO 3) 2,C 4H 6O6, Heavy metals (2.3.13). 2.0 g complies with the limit test for
calculated on the dried basis. heavy metals, Method A (10 ppm).
Description. A white, crystalline powder or colourless crystals. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Identification Loss on drying (2.4.19). Not more than 0.5 per cent, determined
A.To 25 ml of a 0.4 per cent w/v solution add 2 ml of 5 M
Assay. Weigh accurately about 0.3 g, dissolve in 30 ml glacial
ammonia, extract with 20 ml of dichloromethane, filter the
acetic acid. Titrate with 0.1 M perchloric acid, determining
lower layer through anhydrous sodium sulphate and
evaporate to dryness. Place in a freezer for a few minutes to
titration.
congeal the residue and allow to warm to room temperature.
(C15H25NO3)2,C4H6O6.
obtained with metoprolol tartrate RS treated in the same Storage. Store protected from light.
manner or with the reference spectrum of metoprolol.
B. A 5 per cent w/v solution gives reaction B of tartrates (2.3.1).
Metoprolol Tablets
Tests
Metoprolol Tartrate Tablets
(2.4.1), and not more intensely coloured than reference solution Metoprolol Tablets contain not less than 90.0 per cent and
BS8 (2.4.1). not more than 110.0 per cent of the stated amount of metoprolol
tartrate, (C15H25NO3)2, C4H6O6.
Transfer a quantity of the powdered tablets containing about
Related substances. Determine by thin-layer chromatography 40 mg of Metoprolol Tartrate to a separator, add 25 ml of water
(2.4.17), coating the plate with silica gel G. and 4 ml of 5 M ammonia, extract with 20 ml of
763
METRONIDAZOLE IP 2007
dichloromethane, filter the lower layer through anhydrous Metronidazole contains not less than 99.0 per cent and not
sodium sulphate and evaporate to dryness. Place in a freezer more than 101.0 per cent of C6H9N3O3, calculated on the dried
for a few minutes to congeal the residue and allow to warm to basis.
room temperature. Description. A white or yellowish, crystalline powder.
spectrophotometry (2.4.6). Compare the spectrum with that Identification
obtained with metoprolol tartrate RS treated in the same Test A may be omitted if tests B and C are carried out. Tests B
manner or with the reference spectrum of metoprolol. and C may be omitted if test A is carried out.
Tests A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with metronidazole
Dissolution (2.5.2). RS or with the reference spectrum of metronidazole.
Apparatus. No 2 B. When examined in the range 230 nm to 360 nm (2.4.7), a
Medium. 900 ml of simulated gastric fluid (without enzyme) 0.001 per cent w/v solution in 0.1 M hydrochloric acid shows
Speed and time. 100 rpm and 30 minutes. an absorption maximum at about 277 nm and a minimum at
Withdraw a suitable volume of the solution and filter. Dilute about 240 nm; absorbance at about 277 nm, between 0.365
appropriately with the dissolution medium if necessary. and 0.395.
Measure the absorbance (2.4.7) of the solution at the maximum C. Heat about 10 mg in a water-bath with 10 mg of zinc powder,
at about 275 nm. 1 ml of water and 0.25 ml of 2 M hydrochloric acid for 5
Calculate the content of (C15H25NO3)2, C4H6O6 in the medium minutes and cool. The solution gives the reaction of primary
from the absorbance obtained from a solution of known aromatic amines (2.3.1).
concentration of metoprolol tartrate RS.
Tests
D: Not less than 80 per cent of the stated amount of
(C15H25NO3)2, C4H6O6. Appearance of solution. A 5.0 per cent w/v solution in 1 M
hydrochloric acid is not more opalescent than opalescence
Other tests. Comply with the tests stated under Tablets. standard OS2 (2.4.1), and not more intensely coloured than
Assay. Weigh and powder 20 tablets. Weigh accurately a reference solution GYS4 (2.4.1).
quantity of the powder containing about 0.12 g of Metoprolol Related substances. Determine by thin-layer chromatography
Tartrate, transfer to a 100-ml volumetric flask, add about 75 ml (2.4.17), coating the plate with silica gel GF254.
of ethanol (95 per cent) and shake for 15 minutes. Dilute to
volume with ethanol (95 per cent), mix and filter. Dilute 5.0 ml Mobile phase. A mixture of 80 volumes of chloroform, 10
of the filtrate to 50.0 ml with ethanol (95 per cent). Measure volumes of diethylamine, 10 volumes of ethanol (95 per cent)
the absorbance of the resulting solution at the maximum at and 1 volume of water.
about 275 nm (2.4.7). Calculate the content of (C15H25NO3)2, Test solution. Dissolve 0.1 g of the substance under
C4H6O6 from the absorbance obtained by repeating the examination in 10 ml of acetone.
operation using metoprolol tartrate RS in place of the
Metronidazole test solution is not more intense than the spot in the
OH Heavy metals (2.3.13). 1.0 g complies with the limit test for
N heavy metals, Method B (20 ppm).
O2 N CH3
N
C6H9N3O3 Mol. Wt. 171.2 on 1.0 g by drying in an oven at 105° for 3 hours.
Metronidazole is 2-(2-methyl-5-nitro-1H-imidazol-1-yl) Assay. Weigh accurately about 0.15 g, dissolve in 50 ml of
ethanol. anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
764
IP 2007 METRONIDAZOLE BENZOATE ORAL SUSPENSION
acid, determining the end-point potentiometrically (2.4.25). Test solution. Dissolve 0.6 g of the substance under
Carry out a blank titration. examination in 10 ml of equal volumes of chloroform and
1 ml of 0.1 M perchloric acid is equivalent to 0.01712 g of methanol.
C6H9N3O3. Reference solution. A 0.02 per cent w/v solution of the
substance under examination in a mixture of equal volumes of
chloroform and methanol.
dry the plate in air and examine it in ultraviolet light at 254 nm.
Metronidazole Benzoate Any secondary spots in the chromatogram obtained with the
Benzoylmetronidazole test solution are not more intense than the spot in the
O Sulphated ash (2.3.18). Not more than 0.1 per cent.
on 1.0 g by drying at in an oven at 60° at a pressure not
O2 N N CH3 exceeding 0.7 kPa.
N Assay. Weigh accurately about 0.25 g and dissolve in 50 ml of
acetone. Add 10 ml of acetic anhydride. Titrate with 0.1 M
C13H13N3O4 Mol. Wt. 275.3
perchloric acid, determining the end-point potentiometrically
Metronidazole Benzoate is 2-(2-methyl-5-nitro-1H-imidazol- (2.4.25). Carry out a blank titration.
1-yl)ethyl benzoate.
Metronidazole Benzoate contains not less than 98.0 per cent C13H13N3O4.
and not more than 101.0 per cent of C13H13N3O4, calculated on
the dried basis.
Description. A white or cream-coloured, crystalline powder or
flakes. Metronidazole Benzoate Oral
Indentification Suspension
A. When examined in the range 230 nm to 360 nm (2.4.7), a Benzoylmetronidazole Oral Suspension
0.001 per cent w/v solution in ethanol shows an absorption Metronidazole Benzoate Oral Suspension is a suspension of
maximum only at about 309 nm; absorbance at about 309 nm, Metronidazole Benzoate in a suitable aqueous vehicle. It may
about 0.3. contain suitable colouring, flavouring, sweetening, buffering,
B. Gives reaction B of benzoates (2.3.1). suspending and antimicrobial agents.
C. Melting range 98° to 102° (2.4.21). Metronidazole Benzoate Oral Suspension contains not less
Tests stated amount of metronidazole, C6H9N3O3.
pH (2.4.24). 5.0 to 7.0, determined in a 2.0 per cent w/v Identification
suspension.
Extract a quantity of the suspension containing 0.5 g of
Free benzoic acid. Not more than 0.2 per cent, determined by metronidazole with chloroform, filter and evaporate the filtrate
the following method. Dissolve 0.5 g in 25 ml of ethanol to dryness. The residue complies with the following tests.
(95 per cent) and titrate with 0.01 M sodium hydroxide using
phenol red solution as indicator. Carry out a blank titration. A. When examined in the range 230 nm to 360 nm (2.4.7), a
0.001 per cent w/v solution in ethanol shows an absorption
1 ml of 0.01 M sodium hydroxide is equivalent to 0.001221 g maximum only at about 309 nm; absorbance at about 309 nm,
of C7H6O2. about 0.3.
Related substances. Determine by thin-layer chromatography B. Gives reaction B of benzoates (2.3.1).
(2.4.17), coating the plate with silica gel HF254.
Tests
volumes of acetone. Other tests. Complies with the tests stated under Oral Liquids.
765
METRONIDAZOLE INJECTION IP 2007
Assay. Weigh accurately a quantity containing about 200 mg B. Heat 2 ml of the injection in a water-bath for 5 minutes with
of metronidazole, add 10 ml of water and extract with four 10 mg of zinc powder and 0.25 ml of 2 M hydrochloric acid for
quantities, each of 25 ml, of chloroform. Combine the 5 minutes and cool in ice. The solution gives the reaction of
chloroform extracts and wash with two quantities, each of 5 primary aromatic amines (2.3.1).
ml, of water. Wash the aqueous solution with 5 ml of chloroform
and evaporate the combined extracts and washing to dryness Tests
on a water-bath. Add two successive quantities, each of
pH (2.4.24). 4.5 to 7.0.
25 ml, of acetone and 10 ml of acetic anhydride. Titrate with
0.1 M perchloric acid, determining the end-point Bacterial endotoxins (2.2.3). Not more than 0.35 Endotoxin
potentiometrically (2.4.25). Carry out a blank titration. Unit per mg of metronidazole.
1 ml of 0.1 M perchloric acid is equivalent to 0.01712 g of Other tests. Complies with the tests stated under Parenteral
C6H9N3O3. Preparations (Intravenous Infusions).
Determine the weight per ml of the suspension (2.4.29), and Assay. Dilute a suitable volume with sufficient 0.1 M
calculate the content of metronidazole, weight in volume. hydrochloric acid to produce a solution containing 0.001 per
cent w/v of Metronidazole. Measure the absorbance of the
resulting solution at the maximum at about 277 nm (2.4.7),
Labelling. The label states the strength in terms of the using as the blank a solution prepared in the same manner
equivalent amount of metronidazole in a suitable dose-volume. omitting the substance under examination.
Calculate the content of C6H9N3O3 from the absorbance
obtained by repeating the operation using metronidazole RS
in place of the substance under examination.
Metronidazole Injection
Storage. Store protected from light, in single dose containers.
Metronidazole Intravenous Infusion
Labelling. The label states that the contents should not be
Metronidazole Injection is a sterile isotonic solution of used if they contain any visible solid particles.
Metronidazole in Water for Injections. It may contain suitable
buffering agents.
Metronidazole Injection contains not less than 90.0 per cent Metronidazole Tablets
metronidazole, C6H9N3O3. Metronidazole Tablets contain not less than 95.0 per cent and
Description. An almost colourless to pale yellow solution. metronidazole, C6H9N3O3. The tablets may be coated.
A. Determine by thin-layer chromatography (2.4.17), coating A. Shake a quantity of the powdered tablets containing 0.1 g
the plate with silica gel GF254. of Metronidazole with 40 ml of chloroform for 15 minutes,
Mobile phase. A mixture of 70 volumes of chloroform, 28 filter and evaporate the filtrate to dryness.
volumes of methanol, 4 volumes of water and 2 volumes of On the residue, determine by infrared absorption
strong ammonia solution. spectrophotometry (2.4.6). Compare the spectrum with that
Test solution. Dilute a suitable volume of the injection with obtained with metronidazole RS or with the reference spectrum
sufficient mobile phase to produce a solution containing 5 mg of metronidazole.
of Metronidazole per ml. B. Heat a quantity of the powdered tablets containing 10 mg
Reference solution. A 0.5 per cent w/v solution of of Metronidazole in a water-bath with 10 mg of zinc powder,
metronidazole RS in mobile phase. 1 ml of water and 0.25 ml of 2 M hydrochloric acid for
5 minutes and cool in ice. The solution gives the reaction of
primary aromatic amines (2.3.1).
dry the plate in air until the odour of solvent is no longer
detectable and examine in ultraviolet light at 254 nm. The C. Shake a quantity of the powdered tablets containing about
principal spot in the chromatogram obtained with the test 0.2 g of Metronidazole with 4 ml of 0.5 M sulphuric acid and
solution corresponds to that in the chromatogram obtained filter. To the filtrate add 10 ml of picric acid solution and allow
with the reference solution. to stand for 1 hour; the precipitate, after washing with cold
766
IP 2007 MEXILETINE HYDROCHLORIDE
water under suction and drying at 105°, melts at about 150° Mexiletine Hydrochloride
(2.4.21).
CH3 NH2
Tests
O , HCl
CH3
(2.4.17), coating the plate with silica gel GF254. CH3
Mobile phase. A mixture of 80 volumes of chloroform, 20 C11H17NO,HCl Mol. Wt. 215.7
volumes of dimethylformamide and 5 volumes of a 90 per cent
v/v solution of formic acid. Mexiltine Hydrochloride is (RS)-1-methyl-2-(2,6-
xylyloxy)ethylamine hydrochloride.
containing 0.2 g of Metronidazole with 5 ml of a mixture of Mexiletine Hydrochloride contains not less than 99.0 per cent
equal volumes of chloroform and methanol for 5 minutes and and not more than 101.0 per cent of C11H17NO,HCl, calculated
filter. on the anhydrous basis.
Reference solution. A 0.02 per cent w/v solution of 2-methyl- Description. A white or almost white, crystalline powder.
5-nitroimidazole in a mixture of equal volumes of chloroform
Identification
and methanol.
Apply to the plate 20 µl of each solution. After development, A. Determine by infrared absorption spectrophotometry (2.4.6).
dry the plate in air, allow the solvent to evaporate and examine Compare the spectrum with that obtained with mexiletine
in ultraviolet light at 254 nm. Any secondary spot in the hydrochloride RS.
chromatogram obtained with the test solution is not more B. When examined in the range 220 nm to 360 nm (2.4.7), a 0.04
intense than the spot in the chromatogram obtained with the per cent w/v solution in 0.01 M hydrochloric acid shows an
reference solution. absorption maximum at about 260 nm; absorbance at 260 nm,
about 0.46.
Apparatus. No 1
Medium. 900 ml of 0.1 M hydrochloric acid that in the chromatogram obtained with reference solution (b).
D. Dissolve 0.1 g in 3 ml of 0.02 M hydrochloric acid and add
Withdraw a suitable volume of the medium and filter promptly a few crystals of sodium nitrite. Nitrogen is evolved and a
through a membrane filter disc with an average pore diameter yellow colour may be produced slowly.
E. Gives reaction A of chlorides (2.3.1).
and dilute a suitable volume of the filtrate with the same
solvent. Measure the absorbance of the resulting solution at Tests
C6H9N3O3 from the absorbance obtained by repeating the Appearance of solution. A 5.0 per cent w/v solution is clear
operation using metronidazole RS instead of the substance (2.4.1), and colourless (2.4.1).
under examination. pH (2.4.24). 4.0 to 5.5, determined in a 10.0 per cent w/v solution.
D. Not less than 85 per cent of the stated amount of C6H9N3O3. 2,6-Dimethylphenol. Determine by thin-layer chromatography
Other tests. Comply with the tests stated under Tablets. (2.4.17), coating the plate with silica gel G.
Volumes of methanol and 1 volume of strong ammonia
solution.
Metronidazole, transfer to a sintered-glass crucible and extract
with six quantities, each of 10 ml, of hot acetone. Cool, add to Test solution. Dissolve 0.5 g of the substance under
the combined extracts 50 ml of acetic anhydride. Titrate with examination in 10 ml of methanol.
0.1 M perchloric acid, using 0.1 ml of a 1 per cent w/v solution Reference solution. A 0.01 per cent w/v solution of 2,6-
of brilliant green in anhydrous glacial acetic acid as indicator dimethylphenol in methanol.
to a yellowish-green end-point. Carry out a blank titration.
1 ml of 0.1 M perchloric acid is equivalent to 0.01712 g of dry the plate in air, spray with a 0.1 per cent w/v solution of
C6H9N3O3. fast blue BB salt in methanol, dry at about 90° and then spray
767
MEXILETINE CAPSULES IP 2007
with 3 M methanolic potassium hydroxide. Any spot When examined in the range 230 nm to 360 nm (2.4.7), a 0.04
corresponding to 2,6-dimethylphenol in the chromatogram per cent w/v solution of the residue in 0.01 M hydrochloric
obtained with the test solution is not more intense than the acid shows an absorption maximum at 260 nm.
spot in the chromatogram obtained with the reference solution. B. Dissolve 0.1 g of the residue obtained in test A in 3 ml of
Related substances. Determine by thin-layer chromatography 0.02 M hydrochloric acid and add a few crystals of sodium
(2.4.17), coating the plate with silica gel G. nitrite; nitrogen is evolved and a yellow colour may be
Mobile phase. A mixture of 85 volumes of chloroform, 14 produced slowly.
volumes of methanol and 1 volume of strong ammonia C. In the test of Related substances, the principal spot in the
solution. chromatogram obtained with test solution (b) corresponds to
Test solution (a). Dissolve 0.5 g of the substance under that in the chromatogram obtained with reference solution
examination in 10 ml of methanol. (b).
Test solution (b). Dissolve 0.2 g of the substance under D. A 1 per cent w/v solution of the residue obtained in test A
examination in 100 ml of methanol. gives the reactions of chlorides (2.3.1).
Reference solution (a). A 0.025 per cent w/v solution of the Tests
2,6-Dimethylphenol. Determine by thin-layer chromatography
mexiletine hydrochloride RS in methanol.
Apply to the plate 10 µl of each solution. After development, Mobile phase. A mixture of 85 volumes of chloroform, 14
dry the plate in air, spray with ninhydrin solution and heat at volumes of methanol and 1 volume of strong ammonia
110° for 15 minutes. Any secondary spot in the chromatogram solution.
obtained with test solution (a) is not more intense than the Test solution. A 5 per cent w/v solution of the residue obtained
spot in the chromatogram obtained with reference solution (a). in Identification test A in methanol.
Heavy metals (2.3.13). 2.0 g complies with the limit test for Reference solution. A 0.01 per cent w/v solution of 2,6-
heavy metals, Method C (10 ppm). Use 2 ml of lead standard dimethylphenol in methanol.
solution (10 ppm Pb) to prepare the standard.
Sulphated ash (2.3.18). Not more than 0.1 per cent. dry the plate in air, spray with a 0.1 per cent w/v solution of
Water (2.4.19). Not more than 0.5 per cent, determined on 5 g. fast blue BB salt in methanol, dry at about 90° and then spray
with 3 M methanolic potassium hydroxide. Any spot
Assay. Weigh accurately about 0.15 g, dissolve in 50 ml of a corresponding to 2,6-dimethylphenol in the chromatogram
mixture of equal volumes of anhydrous glacial acetic acid obtained with the test solution is not more intense than the
and acetic anhydride, add 10 ml of mercuric acetate solution. spot in the chromatogram obtained with the reference solution.
Titrate with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.4.25). Carry out a blank titration. Related substances. Determine by thin-layer chromatography
C11H17NO,HCl. Mobile phase. A mixture of 85 volumes of chloroform, 14
Volumes of methanol and 1 volume of strong ammonia
solution.
Test solution (a). A 5 per cent w/v solution of the residue
Mexiletine Capsules obtained in Identification test A in methanol.
Test solution (b). Dissolve 20 mg of the residue obtained in
Mexiletine Hydrochloride Capsules
Identification test A in 10 ml of methanol.
Mexiletine Capsules contain not less than 92.5 per cent and Reference solution (a). A 0.025 per cent w/v solution of the
not more than 107.5 per cent of the stated amount of mexiletine residue obtained in Identification test A in methanol.
hydrochloride C11H17NO,HCl.
Identification mexiletine hydrochloride RS in methanol.
A. Shake a quantity of the contents of the capsules containing Apply to the plate 10 µl of each solution. After development,
about 0.5 g of Mexiletine Hydrochloride with 10 ml of methanol, dry the plate in air, spray with ninhydrin solution and heat at
filter, evaporate to dryness and dry the residue at 105°. 110° for 15 minutes. Any secondary spot in the chromatogram
768
IP 2007 MIANSERIN HYDROCHLORIDE
obtained with test solution (a) is not more intense than the Mobile phase. A mixture of 85 volumes of chloroform, 14
spot in the chromatogram obtained with reference solution (a). volumes of methanol and 1 volume of strong ammonia
Dissolution (2.5.2). solution.
Test solution. Dilute a volume of the injection with sufficient
Apparatus. No 1
mobile phase to produce a solution containing 0.2 per cent w/
v of Mexiletine Hydrochloride
injection in methanol.
a membrane filter with an average pore size of 1.0 µm. Measure
the fluorescence intensities using the maximum excitation Reference solution (b). A 0.2 per cent w/v solution of
wavelength at about 265 nm and the maximum emission mexiletine hydrochloride RS in methanol.
wavelength at about 295 nm (2.4.5). Calculate the content of Apply to the plate 10 µl of each solution. After development,
C11H17NO, HCl by comparing the fluorescence intensities dry the plate in air, spray with ninhydrin solution and heat at
obtained with a standard solution of a known concentration 110° for 15 minutes. Any secondary spot in the chromatogram
of mexiletine hydrochloride RS in water. obtained with the test solution is not more intense than the
D. Not less than 80 per cent of the stated amount of C11H17NO, spot in the chromatogram obtained with reference solution (a).
HCl. Other tests. Complies with the tests stated under Parenteral
Assay. Weigh accurately a quantity of the mixed contents of Preparations (Injections).
20 capsules containing about 0.05 g of Mexiletine Assay. To an accurately measured volume containing about
Hydrochloride, mix with 50 ml of 0.01 M hydrochloride acid, 0.125 g of Mexiletine Hydrochloride add sufficient 0.01 M
shake for 30 minutes, dilute to 100.0 ml with 0.01 M hydrochloric acid to produce 250.0 ml and measure the
hydrochloric acid and centrifuge. Measure the absorbance absorbance of the resulting solution at the maximum at about
of the supernatant liquid at the maximum at about 260 nm 260 nm (2.4.7). Calculate the content of C11H17NO, HCI taking
(2.4.7). Calculate the content of C11H17NO,HCl taking 11.6 as 11.6 as the specific absorbance at 260 nm.
the specific absorbance at 260 nm.
Storage. Store in single dose containers.
Mianserin Hydrochloride
Mexiletine Injection H3C
Mexiletine Hydrochloride Injection N
Mexiletine Injection is a sterile solution of Mexiletine
N , HCl
Mexiletine Injection contains not less than 92.5 per cent and
not more than 107.5 per cent of the stated amount of Mexiletine
Hydrochloride C11H17NO, HCI.
C18H20N2,HCl Mol. Wt. 300.8
Identification Mianserin Hydrochloride is (RS)-2-methyl-1,2,3,4,10,14b-
hexahydrodibenzo[c,f] pyrazino[1,2-a]azepine
A. In the test for Related substances, the principal spot in the hydrochloride.
that in the chromatogram obtained with reference solution (b). Mianserin Hydrochloride contains not less than 98.5 per cent
and not more than 101.0 per cent of C18H20N2,HCl, calculated
B. A volume containing 2.5 mg of Mexiletine Hydrochloride on the dried basis.
diluted to 2 ml gives reaction A of chlorides (2.3.1).
Description. A white or almost white crystals or crystalline
Tests powder; odourless or almost odourless.
pH (2.4.24). 5.0 to 6.0. Identification

Related substances. Determine by thin-layer chromatography Test A may be omitted if tests B, C and D are carried out. Tests
(2.4.17), coating the plate with silica gel G. B and C may be omitted if tests A and D are carried out.
769
MIANSERIN TABLETS IP 2007
A. Determine by infrared absorption spectrophotometry (2.4.6). secondary spot in the chromatogram obtained with the test
Compare the spectrum with that obtained with mianserin solution is not more intense than the spot in the chromatogram
hydrochloride RS or with the reference spectrum of mianserin obtained with reference solution (a) and not more than one
hydrochloride. such spot is more intense than the spot in the chromatogram
B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.01
per cent w/v solution shows an absorption maximum only at Sulphated ash (2.3.18). Not more than 0.1 per cent.
about 279 nm; absorbance at about 279 nm, 0.64 to 0.72.
C. Determine by thin-layer chromatography (2.4.17), coating on 1.0 g by drying in an oven at 65° over phosphorus pentoxide
the plate with silica gel GF254. at a pressure not exceeding 0.7 kPa for 5 hours.
Mobile phase. A mixture of 75 volumes of cyclohexane, 20 Assay. Weigh accurately about 0.2 g, dissolve in a mixture of
volumes of ether and 5 volumes of diethylamine. 50 ml of ethanol (95 per cent) and 5 ml of 0.01 M hydrochloric
acid. Titrate with 0.1 M sodium hydroxide, determining the
end-point potentiometrically (2.4.25). Note the volume added
between the two points of inflection.
mianserin hydrochloride RS in dichloromethane.
C18H20N2,HCl.
w/v each of mianserin hydrochloride RS and cyproheptadine
hydrochloride RS in dichloromethane.
phase to rise 15 cm. Dry the plate in a current of cold air for
5 minutes and examine in ultraviolet light at 254 nm. The
Mianserin Tablets
principal spot in the chromatogram obtained with the test Mianserin Hydrochloride Tablets
with reference solution (a). The test is not valid unless the Mianserin Tablets contain not less than 90.0 per cent and not
chromatogram obtained with reference solution (b) shows two more than 110.0 per cent of the stated amount of mianserin
clearly separated principal spots. hydrochloride, C18H20N2, HCl. The tablets may be coated.
D. Gives the reactions of chlorides (2.3.1). Identification
Tests A. Shake a quantity of the powdered tablets containing about

20 mg of Mianserin Hydrochloride with 10 ml of methanol,
pH (2.4.24). 4.0 to 5.5, determined in a 1.0 per cent w/v solution. filter and evaporate the filtrate to dryness.
Related substances. Determine by thin-layer chromatography On the residue, determine by infrared absorption
(2.4.17), coating the plate with silica gel G. spectrophotometry (2.4.6). Compare the spectrum with that
Mobile phase. A mixture of 90 volumes of dichloromethane obtained with mianserin hydrochloride RS or with the
and 10 volumes of methanol. reference spectrum of mianserin hydrochloride.
Test solution. Dissolve 0.2 g of the substance under B. In the Assay, the principal peak in the chromatogram
examination in 10 ml of a mixture of 4 volumes of methanol and obtained with test solution (b) corresponds to the peak due to
1 volume of 10 M ammonia. mianserin in the chromatogram obtained with the reference
solution.
substance under examination in a mixture of 4 volumes of C. The residue obtained in test A gives the reactions of
methanol and 1 volume of 10 M ammonia. chlorides (2.3.1).
Reference solution (b). A 0.002 per cent w/v solution of the Tests
substance under examination in a mixture of 4 volumes of
methanol and 1 volume of 10 M ammonia. Related substances. Determine by thin-layer chromatography
phase to rise 15 cm. Dry the plate in a current of cold air for 5 Mobile phase. A mixture of 90 volumes of dichloromethane
minutes and expose to iodine vapour for 20 minutes. Any and 10 volumes of methanol.
770
IP 2007 MICONAZOLE NITRATE
Test solution. Triturate a quantity of the powdered tablets Miconazole Nitrate

containing 40 mg of Mianserin Hydrochloride with 2 ml of a
mixture of 4 volumes of methanol and 1 volume of strong
ammonia solution and centrifuge. N
Reference solution (a). Dilute 1 volume of the test solution to N
200 volumes with the same solvent mixture.
, HNO
Reference solution (b). Dilute 1 volume of the test solution to O 3
500 volumes with the same solvent mixture.
Cl Cl Cl Cl
dry the plate in a current of cold air for 5 minutes and expose
to iodine vapour for 20 minutes. Any secondary spot in the C18H14Cl4N2O,HNO3 Mol. Wt. 479.2
chromatogram obtained with the test solution is not more Miconazole Nitrate is (RS)-1-[2-(2,4-ichlorophenylmethoxy)-
intense than the spot in the chromatogram obtained with 2-(2,4-dichlorophenyl)ethyl]-1H-imidazole nitrate.
reference solution (a) and not more than one such spot is
Miconazole Nitrate contains not less than 98.5 per cent and
not more than 101.5 per cent of C18H14Cl4N2O,HNO3, calculated
on the dried basis.
Description. A white or almost white, crystalline or micro-
Assay. Determine by gas chromatography (2.4.13). crystalline powder.
Test solution (a). Weigh and powder 20 tablets. Shake a
quantity of the powdered tablets containing about 60 mg of Identification
Mianserin Hydrochloride with 30.0 ml of 0.2 M hydrochloric Test A may be omitted if tests B, C and D are carried out. Tests
acid for 1 hour and filter. To 10.0 ml of the filtrate add 3.0 ml of B, C and D may be omitted if test A is carried out.
1 M sodium hydroxide and 10.0 ml of toluene, mix thoroughly,
centrifuge and use the clear upper layer. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with miconazole
Test solution (b). To 10.0 ml of the filtrate obtained in test nitrate RS or with the reference spectrum of miconazole nitrate.
solution (a) add 3.0 ml of 1 M sodium hydroxide and 10.0 ml of
toluene containing 0.2 per cent w/v of triphenylamine (internal B. When examined in the range 230 nm to 360 nm (2.4.7), a
standard), mix thoroughly, centrifuge and use the clear upper 0.04 per cent w/v solution in a mixture of 90 volumes of 2-
layer. propanol and 10 volumes of 0.1 M hydrochloric acid shows
absorption maxima at about 264 nm, 272 nm and 280 nm; ratio
Reference solution. Add 3.0 ml of 1 M sodium hydroxide and of the absorbance at the maximum at about 272 nm to that at
10.0 ml of toluene containing 0.2 per cent w/v of triphenylamine the maximum at about 280 nm, 1.18 to 1.22.
(internal standard) to 10.0 ml of a solution containing 0.2 per
cent w/v of mianserin hydrochloride RS in 0.2 M C. In the test for Related substances, examine the
hydrochloric acid, mix thoroughly, centrifuge and use the chromatograms obtained in ultraviolet light at 254 nm before
clear upper layer. exposure to iodine vapour. The principal spot in the
Chromatographic system that in the chromatogram obtained with reference solution (a).
silanised diatomaceous support (80 to 100 mesh) coated D. Gives the reactions of nitrates (2.3.1).
with 3 per cent w/w of phenyl methyl silicone fluid
(50 per cent phenyl) (such as OV-17), Tests
– temperature: Appearance of solution. A 1.0 per cent w/v solution in
column 255°, methanol is clear (2.4.1), and not more intensely coloured
inlet port and detector at 240°, than reference solution YS7 (2.4.1).
Inject 1 µl of test solution (a), test solution (b) and the reference (2.4.17), coating the plate with silica gel GF254.
solution.
Mobile phase. A mixture of 60 volumes of hexane, 30 volumes
Calculate the content of C18H20N2, HCl in the tablets. of chloroform, 10 volumes of methanol and 1 volume of strong
Storage. Store protected from light and moisture. ammonia solution.
771
MICONAZOLE CREAM IP 2007
Test solution (a). Dissolve 0.5 g of the substance under 50 ml, of carbon tetrachloride, discarding the organic layers.
examination in 10 ml of a mixture of 90 volumes of methanol Make the aqueous phase alkaline with 2 M ammonia and
and 10 volumes of strong ammonia solution. extract with two quantities, each of 40 ml, of chloroform.
Test solution (b). Dissolve 0.5 g of the substance under Combine the chloroform extracts, shake with 5 g of anhydrous
examination in 100 ml of a mixture of 90 volumes of methanol sodium sulphate, filter and dilute the filtrate to 100 ml with
and 10 volumes of strong ammonia solution. chloroform. Evaporate 50 ml to dryness and dissolve the
residue in 50 ml of a mixture of 90 volumes of methanol and 10
Reference solution (a). A 0.5 per cent w/v solution of volumes of 0.1 M hydrochloric acid.
miconazole nitrate RS in a mixture of 90 volumes of methanol
When examined in the range 230 nm to 360 nm (2.4.7), the
and 10 volumes of strong ammonia solution.
resulting solution shows absorption maxima at about 264 nm,
Reference solution (b). A 0.0125 per cent w/v solution of 272 nm and 280 nm.
miconazole nitrate RS in a mixture of 90 volumes of methanol
and 10 volumes of strong ammonia solution.
Apply to the plate 10 µl of each solution. After development, to miconazole nitrate in the chromatogram obtained with
dry the plate in a current of air for 15 minutes and examine in reference solution (a).
chromatogram obtained with test solution (a) is not more Tests
Other tests. Complies with the tests stated under Creams.
reference solution (b). Expose the plate to iodine vapour until
a brown spot appears in the chromatogram obtained with Assay. Determine by gas chromatography (2.4.13).
reference solution (b) and examine immediately in daylight. Test solution. Weigh accurately a quantity of the cream
Any secondary spot in the chromatogram obtained with test containing about 40 mg of Miconazole Nitrate, mix with 20 ml
solution (a) is not more intense than the spot in the of a mixture of 4 volumes of methanol and 1 volume of 0.5 M
chromatogram obtained with reference solution (b). sulphuric acid and shake with two quantities, each of 50 ml,
Sulphated ash (2.3.18). Not more than 0.1 per cent. of carbon tetrachloride. Wash each organic layer in turn with
the same 10-ml quantity of a mixture of 4 volumes of methanol
Loss on drying (2.4.19). Not more than 0.5 per cent, determined and 1 volume of 0.5 M sulphuric acid. Combine the aqueous
on 1.0 g by drying in an oven at 105° for 2 hours. phase and the washings, make alkaline with 2 M ammonia
Assay. Weigh accurately about 0.4 g, dissolve in 50 ml of and extract with two quantities, each of 50 ml, of chloroform.
anhydrous glacial acetic acid, with slight heating if necessary. To the combined extracts add 10.0 ml of a 0.3 per cent w/v
Titrate with 0.1 M perchloric acid, determining the end-point solution of cetyl palmitate (internal standard) in chloroform
potentiometrically (2.4.25). Carry out a blank titration. and 5 g of anhydrous sodium sulphate, shake, filter, evaporate
the filtrate to a low volume and add sufficient chloroform to
produce 10.0 ml.
C18H14Cl4N2O,HNO3.
miconazole nitrate RS and mix with 10.0 ml of a 0.3 per cent w/
v solution of the internal standard in chloroform and 0.2 ml of
strong ammonia solution, add 1 g of anhydrous sodium
Miconazole Cream sulphate, shake again and filter.
Miconazole Nitrate Cream Reference solution (b). Prepare the solution in the same
manner as reference solution (a) but omitting the addition of
Miconazole Cream contains Miconazole Nitrate in a suitable the internal standard solution.
basis.
Miconazole Cream contains not less than 90.0 per cent and – a glass column 1.5 m x 2 mm, packed with acid-washed,
not more than 110.0 per cent of the stated amount of silanised diatomaceous support (80 to 100 mesh) coated
miconazole nitrate, C18H14Cl4N2O,HNO3. with 3 per cent w/w of phenyl methyl silicone fluid (50
per cent phenyl) (such as 5 per cent OV-101),
Identification
– temperature:
A. Mix a quantity containing 40 mg of Miconazole Nitrate column 270°,
with 20 ml of a mixture of 4 volumes of methanol and 1 volume inlet port and detector at 270°,
of 1 M sulphuric acid and shake with two quantities, each of – flow rate. 30 ml per minute of the carrier gas.
772
IP 2007 MICROCRYSTALLINE CELLULOSE
Inject 1 µl of the test solution, reference solution (a) and methanol and 1 volume of 0.5 M sulphuric acid. Combine the
reference solution (b). aqueous phase and the washings, make alkaline with 2 M
Calculate the content of C18H14Cl4N2O,HNO3 in the cream. ammonia and extract with two quantities, each of 50 ml, of
chloroform. To the combined extracts add 10.0 ml of a 0.3 per
Storage. Store protected from light and moisture at a cent w/v solution of cetyl palmitate (internal standard) in
temperature not exceeding 30°. If it is packed in aluminium chloroform and 5 g of anhydrous sodium sulphate, shake,
tubes, the inner surfaces of the tubes should be coated with a filter, evaporate the filtrate to a low volume and add sufficient
suitable lacquer. chloroform to produce 10.0 ml.
miconazole nitrate RS and mix with 10.0 ml of a 0.3 per cent
w/v solution of the internal standard in chloroform and 0.2 ml
Miconazole Pessaries of strong ammonia solution, add 1 g of anhydrous sodium
Miconazole Nitrate Pessaries; Miconazole Nitrate sulphate, shake again and filter.
Vaginal Tablets; Miconazole Tablets Reference solution (b). Prepare the solution in the same
Miconazole Pessaries contain Miconazole Nitrate in a suitable manner as reference solution (a) but omitting the addition of
basis. the internal standard solution.
Miconazole Pessaries contain not less than 90.0 per cent and Chromatographic system
not more than 110.0 per cent of the stated amount of – a glass column 1.5 m x 2 mm, packed with acid-washed,
miconazole nitrate, C18H14Cl4N2O, HNO3. silanised diatomaceous support (80 to 100 mesh) coated
with 3 per cent w/w of phenyl methyl silicone fluid (50
Identification per cent phenyl) (such as 5 per cent OV-101),
– temperature:
A. Mix a quantity of the crushed pessaries containing 40 mg column 270°,
of Miconazole Nitrate with 20 ml of a mixture of 4 volumes of inlet port and detector at 300°,
methanol and 1 volume of 1 M sulphuric acid and shake with – flow rate. 30 ml per minute of the carrier gas.
two quantities, each of 50 ml, of carbon tetrachloride,
discarding the organic layers. Make the aqueous phase alkaline Inject 1 µl of the test solution, reference solution (a) and
with 2 M ammonia and extract with two quantities, each of reference solution (b).
40 ml, of chloroform. Combine the chloroform extracts, shake Calculate the content of C18H14Cl4N2O, HNO3 in the pessaries.
with 5 g of anhydrous sodium sulphate, filter and dilute the
filtrate to 100 ml with chloroform. Evaporate 50 ml to dryness Storage. Store protected from light and moisture.
and dissolve the residue in 50 ml of a mixture of 90 volumes of
methanol and 10 volumes of 0.1 M hydrochloric acid.
When examined in the range 230 nm to 360 nm (2.4.7), the Microcrystalline Cellulose
resulting solution shows absorption maxima at about 264 nm,
272 nm and 280 nm. Microcrystalline Cellulose is purified, partially
depolymerised cellulose prepared from alpha cellulose.
obtained with the test solution corresponds to the peak due Microcrystalline Cellulose contains not less than 97.0 per cent
to miconazole nitrate in the chromatogram obtained with and not more than 102.0 per cent of cellulose, calculated on
reference solution (a). the dried basis.
Description. A fine or granular, white or almost white powder;
Tests
odourless.
Other tests. Comply with the tests stated under Pessaries.
Identification
A. To about 1 mg add 1 ml of phosphoric acid, heat on a
Test solution. Weigh accurately a quantity of the crushed
water-bath for 30 minutes, add 4 ml of a 0.2 per cent w/v solution
pessaries containing about 40 mg of Miconazole Nitrate, mix
of catechol in phosphoric acid and heat for further 30 minutes;
with 20 ml of a mixture of 4 volumes of methanol and 1 volume
a red colour is produced.
of 0.5 M sulphuric acid and shake with two quantities, each
of 50 ml, of carbon tetrachloride. Wash each organic layer in B. To 50 mg add 2 ml of iodine solution, allow to stand for
turn with the same 10-ml quantity of a mixture of 4 volumes of 5 minutes and remove the excess reagent with the aid of a filter
773
MICROCRYSTALLINE WAX IP 2007
paper and add 1 or 2 drops of sulphuric acid (66 per cent 1 ml of 0.1 M ferrous ammonium sulphate is equivalent to
v/v); a blue-purple colour is produced. 0.000675 g of cellulose.
C. Mix 30 g with 270 ml of water, mix in a blender at 18,000 rpm Storage. Store protected from light and moisture.
for 5 minutes, transfer 100 ml of the mixture to a 100-ml graduated
cylinder and allow to stand for 3 hours. A white, opaque,
bubble-free dispersion is obtained that does not produce a
supernatant liquid.
Microcrystalline Wax
Tests Petroleum Wax (microcrystalline); Amorphous Wax
pH (2.4.24). 5.0 to 7.5, determined on the supernatant liquid Microcrystalline Wax is a mixture of straight-chain,
obtained by shaking 2.0 g with 100 ml of carbon dioxide-free branched-chain and cyclic hydrocarbons, obtained by
water for 5 minutes and centrifuging. solvent fractionation of the still bottom fraction of
Starch and dextrins. Mix 0.1 g with 5 ml of water by vigorous petroleum by suitable dewaxing or de-oiling means.
shaking and add 2 to 3 drops of iodine solution; no blue or Description. A white or cream-coloured, waxy solid; odourless.
brownish-red colour is produced.
Organic impurities. Place 10 mg on a watch-glass and add Tests
0.05 ml of a freshly prepared solution of 0.1 g of phloroglucinol
in 5 ml of hydrochloric acid; no red colour is produced. Acidity or alkalinity. Introduce 35.0 g into a 250-ml separating
funnel, add 100 ml of boiling water and shake vigorously for
Water-soluble substances. Shake 5.0 g with about 80 ml of 5 minutes. Draw off the separated water into a beaker, wash
water for 10 minutes, filter through a filter paper (Whatman further with two quantities, each of 50 ml, of boiling water and
No 42 or equivalent) into a tared beaker and evaporate the add the washings to the liquid in the beaker. To the pooled
filtrate to dryness and dry the residue at 105° for 1 hour. The washings add 0.05 ml of phenolphthalein solution and boil;
residue weighs not more than 10 mg (0.2 per cent). the solution does not acquire a pink colour. Cool, add 0.1 ml of
Arsenic (2.3.10). Mix 5.0 g with 3 g of anhydrous sodium methyl orange solution; no red or pink colour is produced.
carbonate, add 10 ml of bromine solution and mix thoroughly. Solidifying point (2.4.10). 54° to 102°. Follow the method with
Evaporate to dryness on a water-bath, gently ignite and the following modifications. Place in the inner test-tube
dissolve the cooled residue in a mixture of 15 ml of sufficient of the melted substance to fill the tube to a depth of
hydrochloric acid containing 0.15 ml of bromine solution about 50 mm. Stir the substance gently and steadily, without
and 45 ml of water. Add 2 ml of stannous chloride solution scraping the side of the tube, while the tube and its contents
AsT. The resulting solution complies with the limit test for are allowed to cool. The temperature at which the level of the
arsenic (2 ppm). mercury in the thermometer remains stationary for a short time
Heavy metals (2.3.13). 2.0 g complies with the limit test for is taken as the solidifying point.
heavy metals, Method B (10 ppm). Colour. Melt about 10.0 g on a water-bath and pour 5 ml of the
Sulphated ash (2.3.18). Not more than 0.2 per cent. liquid into a clear-glass (15 cm × 16 mm) bacteriological test-
tube; the warm, melted liquid is not more intensely coloured
Loss on drying (2.4.19). Not more than 6.0 per cent, determined than a solution prepared by mixing 3.8 ml of FCS and 1.2 ml of
on 0.5 g by drying in an oven at 105°. CCS (2.4.1), in a similar tube, the comparison being made in
Assay. Weigh accurately about 0.125 g and transfer to a reflected light against a white background, the tubes being
300-ml conical flask with the aid of about 25 ml of water. Add held directly against the background at such an angle that
50.0 ml of 0.083 M potassium dichromate, mix, carefully add there is no fluorescence.
100 ml of sulphuric acid and heat to boiling. Remove from
Organic acids. To 20.0 g add 100 ml of a 50 per cent v/v
heat, allow to stand at room temperature for 15 minutes, cool
solution of ethanol neutralised to phenolphthalein solution
and transfer to a 250-ml volumetric flask. Dilute with water
and titrate rapidly with 0.1 M sodium hydroxide with vigorous
almost to volume, cool to 25°, dilute with water to volume and
agitation, to a sharp pink end-point. Not more than 0.4 ml of
mix. Titrate 50.0 ml of the resulting solution with 0.1 M ferrous
0.1 M sodium hydroxide is required.
ammonium sulphate using 2 to 3 drops of ferroin sulphate
solution as indicator. Repeat the procedure without the Fats, fixed oils and rosin. Digest 10.0 g with 10 ml of 5 M
substance under examination. The difference between the sodium hydroxide at 100° for 30 minutes. Separate the water
titrations represents the amount of ferrous ammonium sulphate layer and acidify with sulphuric acid; no oily or solid matter
required. separates.
774
IP 2007 MINOXIDIL
Ash (2.3.19). Not more than 0.1 per cent, determined on 2.0 g. The principal spot in the chromatogram obtained with the test
It volatilises without emitting an acrid odour. solution corresponds to that in the chromatogram obtained
D. Dissolve about 10 mg in 1 ml of methanol, add 0.1 ml of
cupric sulphate solution; a green colour develops. The
solution becomes greenish-yellow on addition of 0.1ml of 2 M
Minoxidil hydrochloric acid.
Tests
N N NH2 Related substance. Determine by liquid chromatography
(2.4.14).
N
O
NH2 examination in 100 ml of the mobile phase.
C9H15N5O Mol. Wt. 209.3 ml with the mobile phase.
Minoxidil is 2,4-diamino-6-piperidinopyrimidine 3-oxide. Reference solution (b). Dissolve 5 mg of deoxyminoxidil RS
Minoxidil contains not less than 98.5 per cent and not more in the mobile phase and dilute to 20 ml with the mobile phase.
than 101.0 per cent of C9H15N5O, calculated on the dried basis. To 2 ml of this solution add 2 ml of the test solution and dilute
Identification – a stainless steel column 10 cm x 3 mm, packed with
Test A may be omitted if tests B, C and D are carried out. Tests – mobile phase: dissolve 3.0 g of dioctyl sodium
B and C may be omitted if tests A and D are carried out. sulphosuccinate in a mixture of 10 ml of glacial acetic
A. Determine by infrared absorption spectrophotometry (2.4.6). acid and 300 ml of water, adjusted to pH 3.0 with
Compare the spectrum with that obtained with minoxidil RS. perchloric acid and add 700 ml of methanol,
B. Dissolve about 20mg in 0.1 M hydrochloric acid and
dilute to 100 ml with the same solvent (solution A). Dilute 2 ml
of solution A to 100 ml with 0.1 M hydrochloric acid (solution
B) and dilute 2 ml of solution A to 100 ml with 0.1 M sodium Inject the test solution, reference solution (a) and reference
hydroxide (solution C). solution (b). Continue the chromatography for twice the
retention time of the principal peak. In the chromatogram
Measure the light absorption of solutions B and C in the
obtained with the test solution, the sum of the areas of any
range 220 nm to 360 nm (2.4.7). Solution B shows absorption
secondary peaks is not greater than 1.5 times the area of
maxima at about 230 nm and 281 nm; absorbance at about 230
reference solution (a) The test is not valid unless, in the
nm, 0.406 to 0.408 and at about 281 nm, 0.424 to 0.468. Solution
chromatogram obtained with reference solution (c), the
C shows absorption maxima at about 230 nm, 262 nm and
resolution between the peaks corresponding to minoxidil and
288 nm; absorbance at about 230 nm, 0.610 to 0.674, at about
deoxyminoxidil is at least 2.0. Ignore any peak with an area
262 nm, 0.194 to 0.214 and at about 288 nm, 0.222 to 0.242.
less than 0.1 per cent of that of the peak in the chromatogram
C. Determine by thin-layer chromatography (2.4.17), coating obtained with reference solution (a) (0.001 per cent).
Heavy metals (2.3.13).1.0 g complies with the limit test for
Mobile phase. A mixture of 100 volumes of methanol and 1.5 heavy metals, Method C (20 ppm).
volumes of strong ammonia solution.
Test solution. Dissolve 0.1 g of the substance under Loss on drying (2.4.19). Not more than 0.5 per cent, determined
examination in 100 ml of methanol. on 1.0 g by drying in an oven at 105°.
Reference solution. A 0.1 per cent w/v solution of minoxidil Assay. Weigh accurately about 0.15 g, dissolve in 50 ml of
RS in methanol. anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
Apply to the plate 2 µl of each solution. After development, acid, determining the end-point potentiometrically (2.4.25).
dry the plate in air and examine in ultraviolet light at 254 nm. Carry out a blank titration.
775
MINOXIDIL TABLETS IP 2007
1 ml of 0.1 M perchloric acid is equivalent to 0.02093 g of to 10 mg Minoxidil; for tablets containing more than 10 mg of
C9H15N5O. Minoxidil the wavelength used is about 287 nm. Similarly
Storage. Store protected from light and moisture. measure the absorbance of a solution of known concentration
of minoxidil RS. Calculate the content of C9H15N5O.
D. Not less than 75 per cent of the stated amount of C9H15N5O.
Minoxidil Tablets Other tests. Comply with the tests stated under Tablets.
Minoxidil Tablets contains not less than 90.0 per cent and not Assay. Determine by liquid chromatography (2.4.14).
more than 110.0 per cent of the stated amount of minoxidil, Test solution. Weigh and powder 20 tablets. Shake a quantity
C9H15N5O. of the powder containing about 5 mg of Minoxidil with 20 ml
of a solution of medroxyprogesterone acetate (internal
Identification standard) in the mobile phase having a concentration of about
0.2 mg per ml (solution A) for 5 minutes and centrifuge.
A.Transfer a portion of the finely powdered tablets containing
about 10 mg of Minoxidil to a separator, add 25 ml of water, Reference solution. Dissolve an accurately weighed quantity
and extract with three quantities, each of 15 ml, of chloroform. of minoxidil RS in solution A to obtain a solution having a
Combine the chloroform extracts and evaporate with the aid known concentration of about 0.25 mg per ml.
of stream of nitrogen. Wash the inside of the container with
about 5 ml of ethanol (95 per cent), add 0.3 g of potassium
bromide IR and evaporate under vacuum at 50° until dry. The
residue complies with the following test.
– mobile phase: a solution prepared by adding 3.0 g of
Determine by infrared absorption spectrophotometry (2.4.6). docusate sodium per litre to a mixture of 700 volumes of
Compare the spectrum with that obtained with minoxidil RS. methanol, 300 volumes of water and 10 volumes of
B. Determine by thin-layer chromatography (2.4.17), coating glacial acetic acid, adjusting to pH 3.0 with perchloric
the plate with silica gel GF254. acid, filtering and degassing,
Mobile phase. A mixture of 100 volumes of methanol and 1.5 – spectrophotometer set at 254 nm,
volumes of strong ammonia solution. – a 10 µl loop injector.
Test solution. Shake a quantity of the powdered tablets Chromatograph not less than 4 replicate injections of reference
containing 10 mg of Minoxidil with 10 ml of methanol, solution and record the peak response as mentioned below.
centrifuge and use the supernatant liquid. The relative standard deviation is not more than 2.0 per cent
Reference solution. A 0.1 per cent w/v solution of minoxidil and the resolution between the internal standard and minoxidil
RS in methanol. is not less than 2.0.
Apply to the plate 2 µl of each solution. After development, Inject separately the test solution and the reference solution
dry the plate in air and examine in ultraviolet light at 254 nm. and measure the responses for the major peaks. The relative
The principal spot in the chromatogram obtained with the test retention times are about 0.8 for the internal standard and 1.0
solution corresponds to that in the chromatogram obtained for minoxidil.
with the reference solution. Calculated the content of minoxidil, C9H15N5O in the tablets.
Apparatus. No 1
Monothioglycerol
Medium. 900 ml of phosphate buffer pH 7.2
Speed and time. 75 rpm and 15 minutes. Thioglycerol
Withdraw a suitable volume of the medium and filter promptly OH
through a membrane filter disc with an average pore diameter
HO SH
and dilute a suitable volume of the filtrate with the same
C3H8O2S Mol. Wt. 108.2
solvent. Measure the absorbance of the resulting solution at
the maximum at about 231 nm (2.4.7), for tablets containing up Monothioglycerol is 3-mercaptopropane-1,2-diol.
776
IP 2007 MORPHINE AND ATROPINE INJECTION
Monothioglycerol contains not less than 97.0 per cent and B. When examined in the range 230 nm to 360 nm (2.4.7), a
not more than 101.0 per cent of C3H8O2S, calculated on the 0.005 per cent w/v solution in 0.1 M sodium hydroxide shows
anhydrous basis. an absorption maximum only at about 298 nm; absorbance at
Description. A colourless to pale yellow, viscous liquid; odour about 298 nm, about 0.34.
resembling that of sulphides; hygroscopic. C. Add a few mg of the powdered substance to a mixture of
1 ml of sulphuric acid and 0.05 ml of formaldehyde solution;
Tests a purple colour is produced.
pH (2.4.24). 3.5 to 7.0, determined in a 10.0 per cent w/v solution. D. Dissolve 5 mg in 5 ml of water and add 0.15 ml of dilute
Relative density (2.4.29). 1.241 to 1.250. potassium ferricyanide solution and 0.05 ml of ferric chloride
solution; a bluish green colour is produced immediately, which
Refractive index (2.4.27). 1.521 to 1.526. changes rapidly to blue.
Heavy metals (2.3.13). 1.0 g complies with the limit test for E. Gives the reactions of sulphates (2.3.1).
Water (2.3.43). Not more than 5.0 per cent, determined by Acidity. Dissolve 0.2 g in 10 ml of freshly boiled and cooled
Method C on 50.0 g. water and titrate with 0.02 M sodium hydroxide using methyl
Assay. Weigh accurately about 0.2 g, dissolve in 50 ml of red solution as indicator. Not more than 0.2 ml is required to
water and titrate with 0.05 M iodine using 3 ml of starch change the colour of the solution.
solution, added towards the end of the titration, as indicator. Other alkaloids. Not more than 1.5 per cent, calculated on the
1 ml of 0.05 M iodine is equivalent to 0.01082 g of C3H8O2S. dried basis.
Storage. Store protected from light and moisture. Transfer 0.5 g to a separating funnel, add 15 ml of water, 5 ml
of 1 M sodium hydroxide and 10 ml of chloroform, shake,
allow to separate and transfer the chloroform layer to another
separating funnel. Repeat the extraction with two further
Morphine Sulphate quantities, each of 10 ml, of chloroform. Wash the mixed
chloroform solutions with 10 ml of 0.1 M sodium hydroxide
and then with two quantities, each of 5 ml, of water, evaporate
HO to dryness on a water-bath and dry the residue to constant
weight at 105°.
O
, H2SO4, 5H2O Loss on drying (2.4.19). 9.0 to 12.0 per cent, determined on
NCH3
H 0.5 g by drying in an oven at 145° for 1 hour.
HO Assay. Weigh accurately about 0.5 g, dissolve in 30 ml of
2
glacial acetic acid. Titrate with 0.1 M perchloric acid,
(C17H19NO3)2,H2SO4,5H2O Mol. Wt. 758.8
Morphine Sulphate is 7,8-didehydro-4,5α-epoxy-17-
methylmorphinan-3,6α-diol sulphate pentahydrate.
(C17H19NO3)2, H2SO4.
Morphine Sulphate contains not less than 98.0 per cent and
not more than 102.0 per cent of (C17H19NO3)2,H2SO4, calculated
on the dried basis.
Description. White, acicular crystals or cubical masses or a
white, crystalline powder; odourless. Morphine and Atropine Injection
Identification Morphine Sulphate and Atropine Sulphate Injection
A. When examined in the range 230 nm to 360 nm (2.4.7), a Morphine and Atropine Injection is a sterile isotonic solution
0.015 per cent w/v solution shows an absorption maximum in Water for Injections containing 1.0 per cent w/v of Morphine
only at about 285 nm; absorbance at about 285 nm, about 0.65. Sulphate and 0.06 per cent w/v of Atropine Sulphate.
777
MORPHINE INJECTION IP 2007
Morphine and Atropine Injection contains not less than 0.90 of 40, 20 and 20 ml, of a mixture of 1 volume of ethanol (95 per
per cent w/v and not more than 1.10 per cent w/v of morphine cent) and 3 volumes of chloroform, washing each extract with
sulphate, (C17H19NO3)2,H2SO4,5H2O, and not less than 0.054 the same two successive quantities, each of 5 ml, of water and
per cent w/v and not more than 0.066 per cent w/v of atropine continue the extraction until complete extraction of alkaloids
sulphate, (C17H23NO3)2,H2SO4,H2O. has been effected (2.6.4). Evaporate the chloroform, boil the
residue with 10.0 ml of 0.05 M sulphuric acid, cool and titrate
Identification the excess of acid with 0.1 M sodium hydroxide using methyl
red solution as indicator.
plate with silica gel G. 1 ml of 0.05 M sulphuric acid is equivalent to 0.03794 g of
(C17H19NO3)2,H2SO4,5H2O.
Mobile phase. A mixture of 100 volumes of methanol and 1.5
volumes of strong ammonia solution. Storage. Store protected from light.
Test solution. Add 1 ml of 5 M ammonia to 1 ml of the injection,
extract with two quantities, each of 5 ml, of chloroform, filter
the combined extracts through anhydrous sodium sulphate, Morphine Injection
evaporate to dryness in a current of warm air and dissolve the
residue in 0.5 ml of chloroform. Morphine Sulphate Injection
Reference solution (a). Treat 1 ml of a 0.06 per cent w/v Morphine Injection is a sterile solution of Morphine
solution of atropine sulphate RS in the same manner as for Sulphate in Water for Injections.
the test solution. Morphine Injection contains not less than 90.0 per cent and
Reference solution (b). Treat 1 ml of a 1 per cent w/v solution not more than 110.0 per cent of the stated amount of morphine
of morphine sulphate RS in the same manner as for the test sulphate, (C17H19NO3)2,H2SO4,5H2O.
solution.
Identification
dry the plate in air and spray with dilute potassium A. Evaporate a volume containing 5 mg of Morphine Sulphate
iodobismuthate solution. The principal spots in the to dryness on a water-bath. Dissolve the residue in 5 ml of
chromatogram obtained with the test solution correspond to water and add 0.15 ml of dilute potassium ferricyanide
the spots in the chromatograms obtained with reference solution; a bluish green colour is produced immediately, which
solutions (a) and (b). changes rapidly to blue.
B. Gives reaction A of sulphates (2.3.1).
Tests
Other tests. Complies with the tests stated under Parenteral Tests
Preparations (Injections). pH (2.4.24). 2.5 to 6.0.
Assay. For atropine sulphate — To 10.0 ml add 10 ml of water Other tests. Complies with the tests stated under Parenteral
and 5 ml of 1 M sodium hydroxide and extract successively Preparations (Injections).
with 15, 10 and 10 ml of chloroform and continue the extraction
with 10-ml quantities of chloroform until complete extraction
of alkaloids has been effected (2.6.4). Wash the chloroform Test solution. Measure accurately a volume containing about
extracts with the same 5 ml of water (preserve the aqueous 25 mg of Morphine Sulphate and dilute with sufficient of the
solution and the washings for the Assay for morphine mobile phase to produce 100.0 ml, freshly prepared.
sulphate). Evaporate the chloroform, dissolve the residue in Reference solution. Weigh accurately about 25 mg of morphine
2 ml of ethanol (95 per cent), add 2.0 ml of 0.025 M sulphuric sulphate RS and dissolve in sufficient of the mobile phase to
acid, cool and titrate the excess of acid with 0.05 M sodium produce 100.0 ml, freshly prepared.
hydroxide using methyl red solution as indicator.
1 ml of 0.025 M sulphuric acid is equivalent to 0.01737 g of – a stainless steel column 40 cm x 4.6 mm, packed with
(C17H23NO3)2, H2SO4, H2O. octadecylsilane bonded to porous silica (5 µm),
For morphine Sulphate — Combine the aqueous solution – mobile phase: dissolve 0.73 g of sodium heptane-
and washings obtained in the Assay for atropine sulphate, sulphonate in 720 ml of water, add 280 ml of methanol
add 1 g of ammonium sulphate and 25 ml of ethanol (95 per and 10 ml of glacial acetic acid, mix and filter,
cent) and extract with 40 ml, followed by successive quantities – flow rate. 1.5 ml per minute,
778
IP 2007 MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE I
– spectrophotometer set at 280 nm, B. 20 ml gives the reactions of acetates, chlorides phosphates,
– a 20 µl loop injector. sodium salts, potassium salts and magnesium salts (2.3.1).
Inject the test solution and reference solution. Tests
Calculate the content of (C17H19NO3)2, H2SO4, 5H2O in the
injection. pH (2.4.24). 3.0 to 7.0.
Storage. Store protected from light. 5-Hydroxymethylfurfural and Related substances. Dilute a
volume containing 1.0 g of Dextrose to 500.0 ml with water
maximum at about 284 nm; absorbance at about 284 nm, not
more than 0.25 (2.4.7).
Multiple Electrolytes And Dextrose
Injection Type I per ml.
Multiple Electrolytes and Dextrose Injection Type I is a sterile Other tests. Complies with the tests stated under Parenteral
solution of Dextrose and suitable salts in Water for Injection Preparations (Injections).
to provide sodium, potassium, magnesium, acetate, phosphate
and chloride ions. It may contain Hydrochloric Acid or Sodium Assay. For sodium — Dilute suitably with water and determine
Hydroxide used for adjusting the pH. by Method A for flame photometry (2.4.4), or by Method A for
atomic absorption spectrophotometry (2.4.2), measuring at
Composition 589 nm and using sodium solution FP or sodium solution
Sodium acetate 0.32 g AAS respectively, suitably diluted with water for the standard
Potassium chloride 0.13 g solutions.
Dipotassium hydrogen phosphate 0.026 g For total potassium — Dilute suitably with water and
determine by Method A for flame photometry (2.4.4), or by
Magnesium chloride 0.031 g
Method A for atomic absorption spectrophotometry (2.4.2),
Dextrose 5.0 g measuring at 767 nm and using potassium solution FP or
Water for Injections to 100 ml potassium solution AAS respectively, suitably diluted with
water for the standard solutions.
Concentration of electrolytes in mmol / l
Sodium 23.0 For magnesium — To 50.0 ml add 50 ml of water and 5 ml of
strong ammonia-ammonium chloride solution and titrate with
Potassium 20.0
0.005 M disodium edetate using 50 mg of eriochrome black
Magnesium 1.5 T mixture as indicator.
Acetate 23.0 1 ml of 0.005 M disodium edetate is equivalent to 0.1215 mg
Chloride 20.0 of Mg.
Phosphate 1.5 For acetate — Determine by liquid chromatography (2.4.14).
Multiple Electrolytes and Dextrose Injection Type I contains Test solution. Dilute an accurately measured volume of the
not less than 90.0 per cent and not more than 110.0 per cent of preparation under examination quantitatively with water to
the stated amounts of sodium, Na, potassium, K, magnesium, obtain a solution containing about 0.12 per cent w/v of Sodium
Mg, acetate, C2H3O2, and phosphate, PO4. It also contains not Acetate.
less than 90.0 per cent and not more than 120.0 per cent of the
stated amount of chloride, Cl and not less than 90.0 per cent
of sodium acetate in water to obtain a solution having a known
concentration of about 0.12 per cent w/v of sodium acetate.
dextrose, C6H12O6. It contains no antimicrobial agent.
Description. A clear, colourless or faintly straw-coloured
solution.
Identification cross-linked styrene-divinylbenzene copolymer in the
hydrogen form (7 µm) and a guard column 4 cm x 7.8 mm
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution; packed with the same column material,
the solution remains blue and clear. Heat to boiling, a copious – temperature. column 60°,
red precipitate is formed. – mobile phase: 0.1 M sulphuric acid,
779
MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE II IP 2007
– flow rate. 0.8 ml per minute, Multiple Electrolytes and Dextrose

– a 20 µl loop injector. Injection Type II
Inject the reference solution. The test is not valid unless the Multiple Electrolytes and Dextrose Injection Type II is a sterile
tailing factor is not more than 2.0 and the relative standard solution of Dextrose and suitable salts in Water for Injections
deviation for replicate injections is not more than 2.0 per cent. to provide sodium, potassium, calcium, magnesium and
chloride ions. It may contain Hydrochloric Acid or Sodium
Inject separately the test solution and reference solution and
Hydroxide used for adjusting the pH.
measure the responses for the major peak and calculate the
content of acetate in the preparation under examination. Composition
For phosphate — Dilute an accurately measured volume Sodium acetate 0.33 g
containing about 4 mg of phosphate with sufficient water to Sodium chloride 0.088 g
produce 50.0 ml. Transfer 2.0 ml of this solution to a test-tube. Potassium chloride 0.12 g
Add 1.0 ml of a 5 per cent w/v solution of ammonium molybdate
in a cooled mixture of sulphuric acid and water (15:85) and Calcium chloride dihydrate 0.037 g
allow to stand for 3 minutes. Add 1.0 ml of a freshly prepared Magnesium chloride 0.031 g
0.5 per cent w/v solution of hydroquinone containing 1 drop Dextrose 5.0 g
of sulphuric acid and 1.0 ml of a freshly prepared 20 per cent Water for Injections to 100 ml
w/v solution of anhydrous sodium sulphite, mix and allow to
stand for 30 minutes. Measure the absorbance of the resulting Concentration of electrolytes in mmol / l
solution at the maximum at about 640 nm (2.4.7), using as the Sodium 40.0
blank a solution prepared in the same manner by treating 2 ml Potassium 16.0
of water instead of the solution of the preparation under Calcium 2.5
examination. Calculate the content of phosphate from the
absorbance obtained by simultaneously carrying out the Chloride 40.0
operation using a known concentration of about 0.11 mg per Magnesium 1.5
ml of dipotassium hydrogen phosphate in water instead of Acetate 24.0
the solution of the preparation under examination.
Multiple Electrolytes and Dextrose Injection Type II contains
For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of not less than 90.0 per cent and not more than 110.0 per cent of
0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the the stated amounts of sodium, Na, potassium, K, calcium, Ca,
precipitate with water slightly acidified with nitric acid and magnesium, Mg, and acetate, C2H3O2. It contains not less
titrate the excess of silver nitrate with 0.1 M ammonium than 90.0 per cent and not more than 120.0 per cent of the
thiocyanate using ferric ammonium sulphate solution as stated amount of chloride, Cl. It also contains not less than
indicator until a reddish yellow colour is produced. Carry out 90.0 per cent and not more than 110.0 per cent of the stated
a blank titration. amount of dextrose, C6H12O6. It contains no antimicrobial agent.
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total Description. A clear, colourless or faintly straw-coloured
chloride, calculated as Cl. solution.
2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient Identification
water to produce 100.0 ml. Mix well, allow to stand for 30 A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
minutes and determine the optical rotation in a 2-dm tube the solution remains blue and clear. Heat to boiling, a copious
(2.4.22). The observed rotation in degrees multiplied by 0.9477 red precipitate is formed.
represents the weight, in g, of dextrose, C6H12O6 in the volume
taken for assay. B. 20 ml gives the reactions of acetates, chlorides sodium
salts, potassium salts, calcium salts and magnesium salts
Storage. Store in single dose containers at a temperature not (2.3.1).
exceeding 30°.
Labelling. The label states (1) the content of each electrolyte Tests
in terms of millimoles in a given volume; (2) the amount of pH (2.4.24). 3.0 to 7.0.
each ingredient in 100 ml; (3) the total osmolar concentration
in mOsmol per litre; (4) that the preparation should not be 5-Hydroxymethylfurfural and Related substances. Dilute a
used if it contains visible particles. volume containing 1.0 g of Dextrose to 500.0 ml with water
780
IP 2007 MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE III
and measure the absorbance of the resulting solution at the – spectrophotometer set at 210 nm,
maximum at about 284 nm; absorbance at about 284 nm, not – a 20 µl loop injector.
more than 0.25 (2.4.7). Inject the reference solution. The test is not valid unless the
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit tailing factor is not more than 2.0 and the relative standard
per ml. deviation for replicate injections is not more than 2.0 per cent.
Other tests. Complies with the tests stated under Parenteral Inject separately the test solution and the reference solution
Preparations (Injections). and measure the responses for the major peak and calculate
the content of acetate in the preparation under examination.
Assay. For total sodium — Dilute suitably with water and
determine by Method A for flame photometry (2.4.4), or by For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of
Method A for atomic absorption spectrophotometry (2.4.2), 0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the
measuring at 589 nm and using sodium solution FP or sodium precipitate with water slightly acidified with nitric acid and
solution AAS respectively, suitably diluted with water for the titrate the excess of silver nitrate with 0.1 M ammonium
standard solutions. thiocyanate using ferric ammonium sulphate solution as
indicator until a reddish yellow colour is produced. Carry out
For potassium — Dilute suitably with water and determine by a blank titration.
Method A for flame photometry (2.4.4), or by Method A for
atomic absorption spectrophotometry (2.4.2), measuring at 1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total
767 nm and using potassium solution FP or potassium chloride, calculated as Cl.
solution AAS respectively, suitably diluted with water for the For dextrose — To an accurately measured volume containing
standard solutions. 2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient
For calcium — Dilute suitably with water and determine by water to produce 100.0 ml. Mix well, allow to stand for
Method A for flame photometry (2.4.4), or by Method A for 30 minutes and determine the optical rotation in a 2-dm tube
atomic absorption spectrophotometry (2.4.2), measuring at (2.4.22). The observed rotation in degrees multiplied by 0.9477
422.7 nm and using calcium solution FP or calcium solution represents the weight, in g, of dextrose, C6H12O6 in the volume
AAS respectively, suitably diluted with water for the standard taken for assay.
solutions. Storage. Store in single dose containers at a temperature not
For magnesium — To 50.0 ml add 50 ml of water and 5 ml of exceeding 30°.
strong ammonia-ammonium chloride solution and titrate with Labelling. The label states (1) the content of each electrolyte
0.005 M disodium edetate using 50 mg of eriochrome black in terms of millimoles in a given volume; (2) the amount of
T mixture as indicator. each ingredient in 100 ml; (3) the total osmolar concentration
in mOsmol per litre; (4) that the preparation should not be
1 ml of 0.005 M disodium edetate is equivalent to 0.1215 mg
of Mg.
For acetate — Determine by liquid chromatography (2.4.14).
Test solution. Dilute an accurately measured volume of the Multiple Electrolytes and Dextrose
preparation under examination quantitatively with water to Injection Type III
obtain a solution containing about 0.12 per cent w/v of Sodium
Acetate. Multiple Electrolytes and Dextrose Injection Type III is a sterile
solution of Dextrose and suitable salts in Water for Injections
Reference solution. Dissolve an accurately weighed quantity to provide sodium, potassium, acetate, chloride and phosphate
of sodium acetate in water to obtain a solution having a known ions. It may contain Hydrochloric Acid or Sodium Hydroxide
concentration of about 0.12 per cent w/v of sodium acetate. used for adjusting the pH.
Chromatographic system Composition
strong cation-exchange resin consisting of sulphonated Sodium acetate 0.28 g
cross-linked styrene-divinylbenzene copolymer in the Sodium chloride 0.10 g
hydrogen form (7 µm) and a guard column 4 cm × 7.8 mm Potassium chloride 0.15 g
packed with the same column material,
Dipotassium hydrogen phosphate 0.13 g
– mobile phase: 0.1 M sulphuric acid, Dextrose 5.0 g
– flow rate. 0.8 ml per minute, Water for Injections to 100 ml
781
MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE III IP 2007
Concentration of electrolytes in mmol / l For acetate — Determine by liquid chromatography (2.4.14).

Sodium 37.0 Test solution. Dilute an accurately measured volume of the
Potassium 35.0 preparation under examination quantitatively with water to
Acetate 20.0 obtain a solution containing about 0.12 per cent w/v of Sodium
Acetate.
Chloride 37.0
Phosphate 7.5
of sodium acetate in water to obtain a solution having a known
Multiple Electrolytes and Dextrose Injection Type III contains concentration of about 0.12 per cent w/v of sodium acetate.
not less than 90.0 per cent and not more than 110.0 per cent of
the stated amounts of sodium, Na, potassium, K, acetate,
C2H3O2, and phosphate, PO4, and not less than 90.0 per cent
chloride, Cl. It also contains not less than 90.0 per cent and
hydrogen form (7 µm) and a guard column 4 cm x 7.8 mm
not more than 110.0 per cent of the stated amount of dextrose,
packed with the same column material,
Description. A clear, colourless or faintly straw-coloured – mobile phase: 0.1 M sulphuric acid,
solution. – flow rate. 0.8 ml per minute,
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution; Inject the reference solution. The test is not valid unless the
the solution remains blue and clear. Heat to boiling, a copious tailing factor is not more than 2.0 and the relative standard
red precipitate is formed. deviation for replicate injections is not more than 2.0 per cent.
B. 20 ml gives the reactions of acetates, chlorides phosphates, Inject separately the test solution and the reference solution
sodium salts and potassium salts (2.3.1). and measure the responses for the major peak and calculate
the content of acetate in the preparation under examination.
Tests For phosphate — Dilute an accurately measured volume
pH (2.4.24). 3.0 to 7.0. containing about 4 mg of phosphate with sufficient water to
produce 50.0 ml. Transfer 2.0 ml of this solution to a test-tube.
5-Hydroxymethylfurfural and Related substances. Dilute a Add 1.0 ml of a 5 per cent w/v solution of ammonium molybdate
volume containing 1.0 g of Dextrose to 500.0 ml with water in a cooled mixture of sulphuric acid and water (15:85) and
and measure the absorbance of the resulting solution at the allow to stand for 3 minutes. Add 1.0 ml of a freshly prepared
maximum at about 284 nm; absorbance at about 284 nm, not 0.5 per cent w/v solution of hydroquinone containing 1 drop
more than 0.25 (2.4.7). of sulphuric acid and 1.0 ml of a freshly prepared 20 per cent
Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit w/v solution of anhydrous sodium sulphite, mix and allow to
per ml. stand for 30 minutes. Measure the absorbance of the resulting
Other tests. Complies with the tests stated under Parenteral blank a solution prepared in the same manner by treating 2 ml
Preparations (Injections). of water instead of the solution of the preparation under
Assay. For total sodium — Dilute suitably with water and examination. Calculate the content of phosphate from the
determine by Method A for flame photometry (2.4.4), or by absorbance obtained by simultaneously carrying out the
Method A for atomic absorption spectrophotometry (2.4.2), operation using a known concentration of about 0.11 mg per
measuring at 589 nm and using sodium solution FP or sodium ml of dipotassium hydrogen phosphate in water instead of
solution AAS respectively, suitably diluted with water for the the solution of the preparation under examination.
standard solutions. For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of
For potassium — Dilute suitably with water and determine 0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the
by Method A for flame photometry (2.4.4), or by Method A for precipitate with water slightly acidified with nitric acid and
atomic absorption spectrophotometry (2.4.2), measuring at titrate the excess of silver nitrate with 0.1 M ammonium
767 nm and using potassium solution FP or potassium thiocyanate using ferric ammonium sulphate solution as
solution AAS respectively, suitably diluted with water for the indicator until a reddish yellow colour is produced. Carry out
standard solutions. a blank titration.
782
IP 2007 MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE IV
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total Identification

chloride, calculated as Cl.
A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
For dextrose — To an accurately measured volume containing the solution remains blue and clear. Heat to boiling, a copious
2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient red precipitate is formed.
water to produce 100.0 ml. Mix well, allow to stand for 30
minutes and determine the optical rotation in a 2-dm tube B. 20 ml gives the reactions of chlorides ammonium salts,
(2.4.22). The observed rotation in degrees multiplied by 0.9477 sodium salts and potassium salts (2.3.1).
Tests
taken for assay.
Storage. Store in single dose containers at a temperature not pH (2.4.24). 3.0 to 7.0.
exceeding 30°. 5-Hydroxymethylfurfural and Related substances. Dilute a
Labelling. The label states (1) the content of each electrolyte volume containing 1.0 g of Dextrose to 500.0 ml with water
in terms of millimoles in a given volume; (2) the amount of and measure the absorbance of the resulting solution at the
each ingredient in 100 ml; (3) the total osmolar concentration maximum at about 284 nm: absorbance at about 284 nm, not
in mOsmol per litre; (4) that the preparation should not be more than 0.25 (2.4.7).
used if it contains visible particles. Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit
per ml.
Multiple Electrolytes and Dextrose Preparations (Injections).
Injection Type IV Assay. For total sodium — Dilute suitably with water and
determine by Method A for flame photometry (2.4.4), or by
Multiple Electrolytes and Dextrose Injection Type IV is a sterile Method A for atomic absorption spectrophotometry (2.4.2),
solution of Dextrose and suitable salts in Water for Injections measuring at 589 nm and using sodium solution FP or sodium
to provide sodium, potassium, ammonium and chloride ions. solution AAS respectively, suitably diluted with water for the
It may contain Hydrochloric Acid or Sodium Hydroxide used standard solutions.
for adjusting the pH.
For potassium — Dilute suitably with water and determine by
Composition Method A for flame photometry (2.4.4), or by Method A for
Sodium chloride 0.37 g atomic absorption spectrophotometry (2.4.2), measuring at
767 nm and using potassium solution FP or potassium
Potassium chloride 0.13 g
solution AAS respectively, suitably diluted with water for the
Ammonium chloride 0.37 g standard solutions.
Dextrose 5.0 g For ammonium — Transfer an accurately measured volume
Water for Injections to 100 ml of the preparation under examination, containing about 63 mg
Concentration of electrolytes in mmol / l of ammonium, to a 500-ml Kjeldahl flask, dilute to 200 ml with
Sodium 63.0 water, mix and add 50 ml of 40 per cent w/v solution of sodium
hydroxide. Connect the flask immediately to a well-cooled
Potassium 17.0 condenser through a distillation trap. Let the delivery tube
Ammonium 70.0 from the condenser dip into 40 ml of a 4.0 per cent w/v solution
Chloride 150.0 of boric acid contained in a suitable receiver. Heat to boiling
and distil about 200 ml. Cool the liquid in the receiver, if
Multiple Electrolytes and Dextrose Injection Type IV contains
necessary, and titrate with 0.05 M sulphuric acid using methyl
not less than 90.0 per cent and not more than 110.0 per cent of
red solution as indicator. Carry out a blank titration.
the stated amounts of sodium, Na, potassium, K, and
ammonium, NH4 and not less than 90.0 per cent and not more 1 ml of 0.05 M sulphuric acid is equivalent to 1.804 mg of
than 120.0 per cent of the stated amount of chloride, Cl. It also ammonium, NH4.
contains not less than 90.0 per cent and not more than 110.0 For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of
per cent of the stated amount of dextrose, C6H12O6. It contains 0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the
no antimicrobial agent. precipitate with water slightly acidified with nitric acid and
Description. A clear, colourless or faintly straw-coloured titrate the excess of silver nitrate with 0.1 M ammonium
solution. thiocyanate using ferric ammonium sulphate solution as
783
MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE V IP 2007
indicator until a reddish yellow colour is produced. Carry out Multiple Electrolytes and Dextrose Injection Type V contains
a blank titration. not less than 90.0 per cent and not more than 110.0 per cent of
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total the stated amounts of sodium, Na, potassium, K, and
chloride, calculated as Cl. ammonium, NH4 and not less than 90.0 per cent and not more
than 120.0 per cent of the stated amount of chloride, Cl. It also
For dextrose — To an accurately measured volume containing contains not less than 90.0 per cent and not more than
2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient 110.0 per cent of the stated amount of dextrose, C6H12O6. It
water to produce 100.0 ml. Mix well, allow to stand for 30 contains no antimicrobial agent.
minutes and determine the optical rotation in a 2-dm tube
Description. A clear, colourless or faintly straw-coloured
(2.4.22). The observed rotation in degrees multiplied by 0.9477
solution.
taken for assay.
Identification
exceeding 30°. A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
the solution remains blue and clear. Heat to boiling, a copious
Labelling. The label states (1) the content of each electrolyte red precipitate is formed.
in terms of millimoles in a given volume; (2) the amount of
each ingredient in 100 ml; (3) the total osmolar concentration B. 20 ml gives the reactions of acetates, chlorides citrates,
in mOsmol per litre; (4) that the preparation should not be sodium salts, potassium salts, calcium salts and magnesium
used if it contains visible particles. salts (2.3.1).
Tests
pH (2.4.24). 3.0 to 7.0.
Multiple Electrolytes and Dextrose 5-Hydroxymethylfurfural and Related substances. Dilute a
Injection Type V volume containing 1.0 g of Dextrose to 500.0 ml with water
Multiple Electrolytes and Dextrose Injection Type V is a sterile
maximum at about 284 nm; absorbance at about 284 nm, not
solution of Dextrose and suitable salts in Water for Injections
more than 0.25 (2.4.7).
to provide sodium, potassium, calcium, magnesium, acetate,
citrate and chloride ions. It may contain Hydrochloric Acid or Bacterial endotoxins (2.2.3). Not more than 0.5 Endotoxin Unit
Sodium Hydroxide used for adjusting the pH. per ml.
Composition Other tests. Complies with the tests stated under Parenteral
Sodium acetate 0.64 g Preparations (Injections).
Sodium chloride 0.50 g Assay. For total sodium — Dilute suitably with water and
Sodium citrate 0.075 g determine by Method A for flame photometry (2.4.4), or by
Method A for atomic absorption spectrophotometry (2.4.2),
Potassium chloride 0.075 g measuring at 589 nm and using sodium solution FP or sodium
Calcium chloride 0.035 g solution AAS respectively, suitably diluted with water for the
Magnesium chloride 0.031 g standard solutions.
Dextrose 5.0 g For potassium — Dilute suitably with water and determine by
Water for Injections to 100 ml Method A for flame photometry (2.4.4), or by Method A for
atomic absorption spectrophotometry (2.4.2), measuring at
Concentration of electrolytes in mmol / l 767 nm and using potassium solution FP or potassium
Sodium 140.0 solution AAS respectively, suitably diluted with water for the
Potassium 10.0 standard solutions.
Calcium 2.5 For calcium — Dilute suitably with water and determine by
Magnesium 1.5 Method A for flame photometry (2.4.4), or by Method A for
Acetate 47.0 atomic absorption spectrophotometry (2.4.2), measuring at
422.7 nm and using calcium solution FP or calcium solution
Chloride 103.0 AAS respectively, suitably diluted with water for the standard
Citrate 2.5 solutions.
784
IP 2007 MULTIPLE ELECTROLYTES INJECTION TYPE VI
For magnesium — To 50.0 ml add 50 ml of water and 5 ml of – mobile phase: 0.1 M sulphuric acid,
strong ammonia-ammonium chloride solution and titrate with – flow rate. 0.8 ml per minute,
0.005 M disodium edetate using 50 mg of eriochrome black – spectrophotometer set at 210 nm,
T mixture as indicator. – a 20 µl loop injector.
1 ml of 0.005 M disodium edetate is equivalent to 0.1215 mg Inject the reference solution containing 1.0 mg of anhydrous
of Mg. sodium citrate per ml. The test is not valid unless the tailing
For acetate — Determine by liquid chromatography (2.4.14). factor is not more than 1.5 and the relative standard deviation
Test solution. Dilute an accurately measured volume of the
preparation under examination quantitatively with water to Inject separately the test solution and all the three preparations
obtain a solution containing about 0.12 per cent w/v of Sodium of reference solution and measure the responses for the major
Acetate. peak. Plot the responses of all the three preparations of
reference solution versus concentration, in mg of anhydrous
Reference solution. Dissolve an accurately weighed quantity sodium citrate per ml, and draw the straight line best fitting
of sodium acetate in water to obtain a solution having a known the three plotted points. From the graph so obtained, calculate
concentration of about 0.12 per cent w/v of sodium acetate. the content of citrate in mg equivalent to anhydrous sodium
Chromatographic system citrate per litre of the preparation under examination.
– a stainless steel column 30 cm x 7.8 mm, packed with For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of
strong cation-exchange resin consisting of sulphonated 0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the
cross-linked styrene-divinylbenzene copolymer in the precipitate with water slightly acidified with nitric acid and
hydrogen form (7 µm) and a guard column 4 cm x 7.8 mm titrate the excess of silver nitrate with 0.1 M ammonium
packed with the same column material, thiocyanate using ferric ammonium sulphate solution as
– column. temperature 60°, indicator until a reddish yellow colour is produced. Carry out
– mobile phase: 0.1 M sulphuric acid, a blank titration.
– spectrophotometer set at 210 nm, 1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total
– a 20 µl loop injector. chloride, calculated as Cl.
Inject the reference solution. The test is not valid unless the For dextrose —To an accurately measured volume containing
tailing factor is not more than 2.0 and the relative standard 2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient
deviation for replicate injections is not more than 2.0 per cent. water to produce 100.0 ml. Mix well, allow to stand for 30
minutes and determine the optical rotation in a 2-dm tube
Inject separately the test solution and the reference solution (2.4.22). The observed rotation in degrees multiplied by 0.9477
and measure the responses for the major peak and calculate represents the weight, in g, of dextrose, C6H12O6 in the volume
the content of acetate in the preparation under examination. taken for assay.
For citrate — Determine by liquid chromatography (2.4.14). Storage. Store in single dose containers at a temperature not
Test solution. Preparation under examination. exceeding 30°.
Reference solution. Dissolve an accurately weighed quantity Labelling. The label states (1) the content of each electrolyte
of anhydrous sodium citrate, previously dried at 180° for 18 in terms of millimoles in a given volume; (2) the amount of
hours, in water to obtain a stock solution having a known each ingredient in 100 ml; (3) the total osmolar concentration
concentration of about 10 mg per ml. Dilute accurately in mOsmol per litre; (4) that the preparation should not be
measured volumes of this solution quantitatively with water used if it contains visible particles.
to obtain three solutions having known concentrations of
about 0.5, 1.0 and 2.0 mg, respectively of anhydrous sodium
citrate per ml.
Chromatographic system Multiple Electrolytes Injection Type VI
strong cation-exchange resin consisting of sulphonated Multiple Electrolytes Injection Type VI is a sterile solution of
cross-linked styrene-divinylbenzene copolymer in the suitable salts in Water for Injections to provide sodium,
hydrogen form (7 µm) and a guard column 4 cm × 7.8 mm potassium, calcium, magnesium, acetate, citrate and chloride
packed with the same column material, ions. It may contain Hydrochloric Acid or Sodium Hydroxide
– column. temperature 60°, used for adjusting the pH.
785
MULTIPLE ELECTROLYTES AND DEXTROSE INJECTION TYPE II IP 2007
– flow rate. 0.8 ml per minute, Multiple Electrolytes and Dextrose

– a 20 µl loop injector. Injection Type II
Inject the reference solution. The test is not valid unless the Multiple Electrolytes and Dextrose Injection Type II is a sterile
tailing factor is not more than 2.0 and the relative standard solution of Dextrose and suitable salts in Water for Injections
deviation for replicate injections is not more than 2.0 per cent. to provide sodium, potassium, calcium, magnesium and
chloride ions. It may contain Hydrochloric Acid or Sodium
Inject separately the test solution and reference solution and
Hydroxide used for adjusting the pH.
measure the responses for the major peak and calculate the
content of acetate in the preparation under examination. Composition
For phosphate — Dilute an accurately measured volume Sodium acetate 0.33 g
containing about 4 mg of phosphate with sufficient water to Sodium chloride 0.088 g
produce 50.0 ml. Transfer 2.0 ml of this solution to a test-tube. Potassium chloride 0.12 g
Add 1.0 ml of a 5 per cent w/v solution of ammonium molybdate
in a cooled mixture of sulphuric acid and water (15:85) and Calcium chloride dihydrate 0.037 g
allow to stand for 3 minutes. Add 1.0 ml of a freshly prepared Magnesium chloride 0.031 g
0.5 per cent w/v solution of hydroquinone containing 1 drop Dextrose 5.0 g
of sulphuric acid and 1.0 ml of a freshly prepared 20 per cent Water for Injections to 100 ml
w/v solution of anhydrous sodium sulphite, mix and allow to
stand for 30 minutes. Measure the absorbance of the resulting Concentration of electrolytes in mmol / l
solution at the maximum at about 640 nm (2.4.7), using as the Sodium 40.0
blank a solution prepared in the same manner by treating 2 ml Potassium 16.0
of water instead of the solution of the preparation under Calcium 2.5
examination. Calculate the content of phosphate from the
absorbance obtained by simultaneously carrying out the Chloride 40.0
operation using a known concentration of about 0.11 mg per Magnesium 1.5
ml of dipotassium hydrogen phosphate in water instead of Acetate 24.0
the solution of the preparation under examination.
Multiple Electrolytes and Dextrose Injection Type II contains
For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of not less than 90.0 per cent and not more than 110.0 per cent of
0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the the stated amounts of sodium, Na, potassium, K, calcium, Ca,
precipitate with water slightly acidified with nitric acid and magnesium, Mg, and acetate, C2H3O2. It contains not less
titrate the excess of silver nitrate with 0.1 M ammonium than 90.0 per cent and not more than 120.0 per cent of the
thiocyanate using ferric ammonium sulphate solution as stated amount of chloride, Cl. It also contains not less than
indicator until a reddish yellow colour is produced. Carry out 90.0 per cent and not more than 110.0 per cent of the stated
a blank titration. amount of dextrose, C6H12O6. It contains no antimicrobial agent.
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total Description. A clear, colourless or faintly straw-coloured
chloride, calculated as Cl. solution.
2 g to 5 g of Dextrose, add 0.2 ml of 5 M ammonia and sufficient Identification
water to produce 100.0 ml. Mix well, allow to stand for 30 A. To 1 ml add 0.05 ml of potassium cupri-tartrate solution;
minutes and determine the optical rotation in a 2-dm tube the solution remains blue and clear. Heat to boiling, a copious
(2.4.22). The observed rotation in degrees multiplied by 0.9477 red precipitate is formed.
taken for assay. B. 20 ml gives the reactions of acetates, chlorides sodium
salts, potassium salts, calcium salts and magnesium salts
Storage. Store in single dose containers at a temperature not (2.3.1).
exceeding 30°.
Labelling. The label states (1) the content of each electrolyte Tests
in terms of millimoles in a given volume; (2) the amount of pH (2.4.24). 3.0 to 7.0.
each ingredient in 100 ml; (3) the total osmolar concentration
in mOsmol per litre; (4) that the preparation should not be 5-Hydroxymethylfurfural and Related substances. Dilute a
used if it contains visible particles. volume containing 1.0 g of Dextrose to 500.0 ml with water
780
IP 2007 MUSTINE INJECTION
Chromatographic system Mustine Hydrochloride contains not less than 98.0 per cent
– a stainless steel column 30 cm x 7.8 mm, packed with and not more than 101.0 per cent of C5H11CI2N, HCl.
strong cation-exchange resin consisting of sulphonated Description. A white or almost white, crystalline powder or
cross-linked styrene-divinylbenzene copolymer in the mass; hydroscopic; vesicant.
hydrogen form (7 µm) and a guard column 4 cm × 7.8 mm
packed with the same column material, Identification
– mobile phase: 0.1 M sulphuric acid, A. Dissolve 50 mg in 5 ml of water and add 1 ml of 5 M sodium
– flow rate. 0.8 ml per minute, hydroxide; oily globules are produced which dissolve on
– spectrophotometer set at 210 nm, warming.
– a 20 µl loop injector. B. Dissolve 50 mg in 5 ml of water and add 0.02 ml of potassium
Inject the reference solution containing 1.0 mg of anhydrous mercuri-iodide solution; a cream-coloured precipitate is
sodium citrate per ml. The test is not valid unless the tailing produced.
factor is not more than 1.5 and the relative standard deviation A. Melts at about 108° (2.4.21).
Inject separately the test solution and all the three preparations Tests
of reference solution and measure the responses for the major Assay. Weigh accurately about 0.2 g, add 15 ml of 1 M ethanolic
peak. Plot the responses of all the three preparations of potassium hydroxide and 15 ml of water and boil under a
reference solution versus concentration, in mg of anhydrous reflux condenser for 2 hours. Evaporate the solution to half its
sodium citrate per ml, and draw the straight line best fitting volume on water-bath, dilute to 150 ml with water, add 3 ml of
the three plotted points. From the graph so obtained, calculate nitric acid and 50.0 ml of 0.1 M silver nitrate. Shake vigorously
the content of citrate in mg equivalent to anhydrous sodium and filter. Wash the residue with water and titrate the excess
citrate per litre of the preparation under examination. of silver nitrate in the combined filtrate and washings with
For total chloride — To 20.0 ml add 30 ml of water, 50.0 ml of 0.1 M ammonium thiocycanate using 1 ml of ferric ammonium
0.1 M silver nitrate and 2 ml of nitric acid. Filter, wash the sulphate solution as indicator.
precipitate with water slightly acidified with nitric acid and
titrate the excess of silver nitrate with 0.1 M ammonium
C5H11CI2N, HCl.
thiocyanate using ferric ammonium sulphate solution as
indicator until a reddish yellow colour is produced. Carry out Storage. Store protected from light and moisture at a
a blank titration. temperature not exceeding 30°.
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total Labeling. The label states that the contents of the container
chloride, calculated as Cl. are strongly vesicant.
exceeding 30°.
Labelling. The label states (1) the content of each electrolyte Mustine Injection
in terms of millimoles in a given volume; (2) the amount of
each ingredient in 100 ml; (3) the total osmolar concentration Mustine Hydrochloride Injection
in mOsmol per litre; (4) that the preparation should not be
Mustine Injection is a sterile material consisting of Mustine
Hydrochloride with or without buffering agents and other
excipients. It is filled in a sealed container.
Mustine Hydrochloride The injection is constituted by dissolving the contents of the

Nitrogen Mustard Injections or Sodium Chloride Intravenous Infusion,
immediately before use.
CH3
N , HCl The constituted solution complies with the requirements for
Cl Cl Clarity of solution and Particulate matter stated under
C5H11CI2N,HCl Mol Wt. 192.4 Parenteral Preparations (Injections).
Mustine Hydrochloride is bis(2-chloroethyl)methylamine Storage. The constituted solution deteriorates rapidly on
hydrochloride. storage and should be used immediately after preparation.
787
MUSTINE INJECTION IP 2007
Mustine Injection contains not less than 90.0 per cent and not containers, containing 40 mg of Mustine Hydrochloride,
more than 110.0 per cent of the stated amount of the stated dissolve in 10 ml of ethanol (95 per cent), previously
amount of mustine hydrochloride, C5H11CI2N,HCl. neutralised to dilute phenolphthalein solution. Titrate with
The contents of the sealed container complies with the tests 0.01 M sodium hydroxide using dilute phenolphthalein
stated under Parenteral Preparations (Powders for solution as indicator.
Injection) and with the following requirements. 1 ml of 0.01 M sodium hydroxide is equivalent to 0.001925 g
of C5H11CI2N, HCl.
Identification
Dissolve about 20 mg in 1 ml of water and add 0.02 ml of exceeding 30°.
potassium mercuri-iodide solution; a cream-coloured Labelling. The label states (1) that the contents are strongly
precipitate is produced. vesicant; (2) the amount of Mustine Hydrochloride in the
Tests container, (3) that the injection should be used immediately
after preparation.
Weigh accurately a quantity of the mixed contents of the ten
788

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INDIAN

THE INDIAN PHARMACOPOEIA COMMISSION

General Monographs on Dosage Forms .................................................................

Monographs on Drug Substances, Dosage Forms and Pharmaceutical Aids

General Statements ....

Meanings of Terms — per cent v/v (percentage, volume in volume) expressing

General requirements ....

Creams Sterility. When the cream is labelled as sterile, it complies

not cause deterioration of the preparation as a result of Eye Ointments

Dimensions in millimeters (tolerances ± 1 mm, unless otherwise specified)

Fig. 3: Apparatus B. Anderson cascade impactor

respirable powder. It is connected to the induction port as Procedure

Dimensions in millimeters unless otherwise stated

(Dimensions are in millimeters unless otherwise stated)

Table 4 – Specifications of the apparatus shown in Fig. 6

Table 5 – Component specification for set-up in Fig. 7

Fig. 7: Experimental set-up for testing powder inhalers

Oral Powders determination using another 20 containers taken at random.

Tests Shell Pessaries. Shell Pessaries, also known as Vaginal

If coated tablets fail to comply because of adherence to the Enteric-coated Tablets

Tests Prolonged- release Tablets

Soluble Tablets Tablets for Use in the Mouth

DRUG SUBSTANCES, DOSAGE FORMS

Aluminium Acetate Ear Drops ....

Amoxycillin Dispersible Tablets ....

Atorvastatin Tablets ....

Glacial Acetic Acid 1 ml of 1 M sodium hydroxide is equivalent to 0.06005 g of

N Reference solution (c). Dissolve 10 mg of adenine RS and 10

Storage. Store protected from light in containers preferably Chromatographic system

B. Extract a quantity of the powdered tablets containing 100 Tests

Chromatographic system Calculate the content of C17H13ClN4 in the tablet.

Identification Aluminium Acetate Ear Drops

Identification Storage. Store protected from moisture.

Tests Aluminium Sulphate

Identification hydrochloride RS or with the reference spectrum of ambroxol

mechanical means for 30 minutes. Dilute with 0.1 M Tests

Aminocaproic Acid Tablets O

Triturate 2 tablets with 10 ml of water and filter into 100 ml of

Identification Calculate the amount of C2H8N2 present for each g of C7H8N4O2

1). To 15 ml of solution A add 1 ml of 0.1 M hydrochloric acid, Tests

Amitriptyline Hydrochloride Mobile phase. A mixture of 85 volumes of cyclohexane,

Identification than the spot in the chromatogram obtained with reference

Reference solution (c). Dissolve 5 mg of the substance under Identification

Related substances. Determine by thin-layer chromatography Tests

Speed and time. 100 rpm and 60 minutes. Amoxycillin Injection

Reference solution. Dissolve an accurately weighed quantity Identification

Chromatographic system Solvent mixture. Dissolve 6.8 g of monobasic potassium

Ampicillin Sodium is sodium (6R)-6-(α-phenyl-D-glycyl- – inlet port and detector.150°,

Bacterial endotoxins (2.2.3). Not more than 0.15 Endotoxin Tests

Ampicillin Injection Determine by gas chromatography (2.4.13).

Reference solution (b). Dissolve caffeine in reference solution Identification

digesting about 10 mg of dry soluble maize or corn starch). Tests

Identification NOTE — 15 ml of solution A or 25 ml of solution B are to be

A. To 2 ml of the filtrate, add 5 ml of ethanol (95 per cent) and Identification

50 volumes of glacial acetic acid, 1 volume of sulphuric acid Chromatographic system

Tests B. To a volume containing 40 mg of Ascorbic Acid add 4 ml of

C. To 1 ml of solution A, add about 0.1 ml of 2 M nitric acid Tests

Inject 20 µl of the test solution and reference solution (b), Identification

Atenolol is (RS)-4-(2-hydroxy-3- Reference solution (b). Dissolve 50 mg of atenolol impurity

peak in the chromatogram obtained with reference solution Atropine Methonitrate

bubbles disappear. Add 1 ml of á-naphthol solution. A pale- Identification

Benzylpenicillin Injection ....