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Microbial contamination of pharmaceutical preparations :

Components of pharmaceuticals and cosmetics which encourage

microbial growth include:

1\ Thickening and suspending agents such as gums : ( e.g. acacia

,tragacanth).

2\ Starches cellulose derivatives

3\ High sugar content in syrups although inhibits the growth of many

microorganisms due to it is high osmotic pressure, yet osmo-tolerant
moulds and yeasts not only can survive but they able to ferment this
sugar and flourish in it.

4/ Proteins and vitamins in cosmetics and toiletary preparations highly

nutritive to microorganisms and May inactivate preservatives and even
act as a source of contamination.

5 / The non ionic and to a lesser extent anionic surfactant which are
commonly used as solubilizing agents and emulgents support the
growth of certain contaminants .e.g certain strains of Pseudomonas
aeruginosa , penicillium. nonatum , Candida .albicans , Aspergillus niger
where we found capable of decomposing and utilizing tweens (Non-
ionic) , alkyl sulphate, sulphonated fatty acids.

6/ Water in Pharmaceutical preparation is essential for metabolism and

multiplication of all organism

N:B:-

Certain bacteria and some fungi can survive for extended periods of
time in anhydrous products e.g: ointments, oils.

Few organisms can even metabolize and use of some oils and fats as a
carbon source e.g : certain species of Aspergillus and Penicillium can
attack a rachis oil and liquid paraffin respectively.

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Important consequences of microbial contamination:

Contaminants are usually classified into:

1) Primary pathogens can infect humans under normal resistant.

2) Secondary pathogens or opportunistics.

3) Non pathogens.

Microorganisms may affect either the consumer , the preparation or

both depending on the class of contaminant

1) Effect on the consumer may cause infection, may liberate toxins ,

other metabolic products such as organic acids and amines may be
produced by contaminant which may exert irritating , sensitizing or
other allergic conditions in the consumer

2) Effect on the preparation: contaminant particularly non pathogens

possess metabolic reactions such as oxidation , reduction , hydrolysis ,
decarboxylation, deamination and dehydration .These may lead to
inactivation of the active constituent of the preparation e.g penicillins by
penicillinase enzyme .

a) Destruction of preservatives , disinfectants and other antimicrobial

agents .

b) Microorganisms may affect physiochemical properties of various

preparations through their degradation of various auxiliary materials
present .

c) Rancidity of ointments, creams , discoloration of tablets, unpleasant

aroma and flavour in syrups etc.

d) The breakdown of emulsions may be due to upset in phase

equilibrium as result in hydrolysis in oil phase ,the alteration of pH of
aqueous phase or decomposition of the emulgent.

e) Changes in aroma in pharmaceutical or Cosmetics products due to

contaminants e.g: production of hydrogen sulphide (H2S) , fishy odor of
amines.

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f) Occurrence of visible microbial growth objectionable from consumer's
point of view e.g the occurance of visible microbial growth particularly
fungi on the surface of solid or semi-solid preparation.

g) Colour changes in pharmaceutical preparations may be caused by

change of pH or redox potential of the components excreted by
microbial growth.

Members of genus Pseudomonas they can produce soluble pigments

color change to blue green ( exo- pigment ) .

Spoilage organisms can alter the following :-

1] The aesthetic qualities such as smell, taste , appearance .

2] Physical properties (pH, Viscosity).

3] Efficacy of the product.

4] Production of toxins .

3) Effect on the manufacturing company :

The presence of microorganisms in Pharmaceutical products forces

consumers to lose faith in the manufacturing company and this may
cause the company considerable financial defeate.

Sources and control of microbial contamination:

A) Contamination during manufacturing :

The main source of microorganisms before or during formulation and

manufacture include the following

1) Raw Materials :

The degree of microbial contamination of raw materials depends on

their origin.

Generally those from animal e.g. gelatin, thyroid or vegetable e.g:

gums, starches ,and powdered plants origin are heavily contaminated
with variable organisms.

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 Solid natural materials as Kaolin, bentonite and talc contain anaerobic
spore bearing bacteria, moulds and other organisms.

On other hand synthetic or semi synthetic ingredients usually contain

quite low microbial counts.

All raw materials should be kept under Proper Storage conditions to

guard against their contaminations and to discourage any increase in
microbial population.

Before manufacture, the raw material should be checked for viable

organisms and for the presence of specific pathogens such as
Salmonella, E.coli , Staphylococcus aureus , and Pseudomonas
aeruginosa .

Heavily contaminated raw materials are either condemned if cheap or

else sterilized by gaseous or heating process before formulation.

2) water supply

Types of water:

1} Raw or mains water

2} Softened water: Prepared by removal of calcium , magnesium , and

certain other metal cations in hard water.

Softened water is often used for washing containers and for cooling
systems.

3} Deionized or demineralized water: It is prepared by passing mains

water through anion and cation exchange resin bed to remove the ions.

Deionized water is used in pharmaceutical formulations ,for washing

containers and plant, and for the preparation of disinfectant solution.

4} Distilled water

5} Water produce by reverse osmosis : Is forced by an osmotic pressure

through a semi-permeable membrane which acts a molecular filter . The
diffusion of solutes dissolved in the water is impeded , and those with a
molecular weight in excess of 250 do not diffuse at all. The process,
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which is the reverse of the natural process of osmosis , thus removes
M.Os and their pyrogens .

In pharmaceutical formulation water is a source of many potential

harmful organisms and at the same time provide the vehicle for growth
of organisms introduced by other Sources without effective treatment to
minimize contamination. Water can within a few days contain large
number of Gram-negative bacilli , wide variety of bacteria, molds and
yeasts.

Various methods have been suggested for controlling the contamination

of water supply includeing Filtration , or Treatment with UV radiation
and chemical disinfectants or the use of freshly distilled water under
rigid hygienic conditions .

3) Air Supply

Act as a critical source of contamination. In order to eliminate or at least

minimize such contaminants , one or more of the following measures
maybe followed :

A) The intake of air into manufacturing space could be made through

filters and conditions which remove most of the dust particles together
with majority of organisms.

All filters should be periodically cleaned and disinfected.

Chemical disinfection also can be used to reduce the microbial count.

B) The air currents could be exposed for UV radiation for more

disinfection.

C) Though the building dust should be controlled by avoiding dry

cleaning and sweeping of walls and floors.

The use of solutions of soaps and disinfectaant is of value.

D) The building of the firm should be located away from crowded

Industrial area generating dust and fumes.

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E) The floors, walls and ceiling of the plant should be constructed from
non porous smooth materials having no cracks and as few Services as
possible to minimize dust collection and to allow for frequent cleaning
with detergent and water.

4) Equipment

Apparatus and machines used in the manufacture of pharmaceutical and

cosmetics may possess small orifices cervices , cross-connections, dead
end lines , U,S, or T turns .

All such arrangements have difficult access and therefore potential

traps for microorganisms wish might grow in the remainings of the
product left between successive batches .

It is very important to eliminate such microbial reservoir and preventing

them from contaminating the subsequent product , The equipment
therefore must be designed so that they can be thoroughly cleaned with
detergent and water such cleaning is essential immediately after each
use.

5) Employees:

Human beings may a harbour various microorganisms in their

respiratory tract, intestinal tract , and on their skin or their hair.
Consequently they constitute the most important source of microbial
contamination, since they are sheding microorganisms during talking ,
laughing, coughing or sneezing near the pharmaceuticals being
prepared. They may also contaminate these products through handling if
they do not follow strict personal hygienic measures.
The employees should wear clean uniforms and approved head
covering at all times . Individual with obvious colds, fever and those
with visible hands or face infection either not to be allowed to handle
the product or be made to wear adequate masks or gloves .The
personnel should receive periodic medical examination in order to
discover early infection or carrier state.

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B) Contamination after manufacture :

The liners of paper and cork and the work closure often contain many
microorganisms therefore may act as source of contamination. Plastic
closures are free from these defects. All types of cork and plastic
closures may absorb or inactivate preservatives in the formulation and
so they encourage microbial contamination. In addition plastic suffer
from other disadvantages. They are porous to varying degrees , and
some allow the diffusion of oxygen (O2) and CO2 and may thus facilitate
microbial growth in the packed products.

The finished products may be contaminated during consumption when

the consumer opens container, its contents are immediately exposed to
air borne microorganisms , such a problem is significant in the case of
multi-dose Preparations.

Dispensing creams and ointments in wide mouth jars invites the

consumer to use his fingers thus producing more contaminating
organisms.
The use of collapsiable tubes instead of the jars are eliminates this
hazard and Lowers the level of contamination during the consumption of
product.

The use of droppers in the case of ophthalmic ear, nose liquid

preparations introduces after each application a number of
microorganisms including some pathogens. This is very important since
most of such preparations are available in multi-dose forms and they are
often used by more than one member in the family of the consumer .

In some cosmetic such as lipstick ,eye mascara ,solid cake makeup,

microorganisms are most likely to be produced during application of
such products through breath, saliva, via the brush or by the use of the
buffs.
The control measure designed for minimizing the extent of
contamination of products during the consumption are of two types:

A) The formulation of products in single doses particularly in the eye

drops.

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B) Incorporation of preservatives in the preparation prevents the
contamination after manufacture and during consumption however
preservation should not be substituted for high level of hygiene and
sanitation .

This is important in the light of observations that some organisms resist

the action of certain preservatives and even destroy them.

Microbiological control of non sterile pharmaceutical products :

The majority of products intended for topical use and oral

administration cannot be free from viable microorganisms. However
there is recognized need for microbiological control of such non sterile
products.

Two tests for the quality control of non-sterile products:

1) The product should be selectively free from those microorganisms

which can cause disease when they gain access to the body by the route
of administration recommended for the product, thus Salmonella and
E.coli as indicator of fecal pollution , should be absent in preparations to
be taken orally.

P.aeruginosa should not be present in products to be used in or near

the eye , and S.aureus in formulations to be applied on skin.

2) In considering the level of contamination with non-pathogenic

organisms the total number which may be present would vary with each
product , and from one manufacturer to another e.g. the British
pharmacopoeia for a medicine to be administered orally, there shouldn't
be more than 103 aerobic bacteria or 102 fungi/gm or cm3 of products.

*Higher levels may be permissible if the products contain raw materials

of natural origin e.g. U.S.P recommended that gelatin should not contain
more than 5,000 viable cells/gm with no E.coli 10 gm sample.

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