IMPERIAL ACRICUL1URAL

RESEARCH INSTITUTE, NEW DELHI.

MeGRAW-HILL PUBLICATIONS IN PHAIlMACY

QlTAN'ITI'ATIVE PHARIV[ACEUTICAL
CHEMISTRY

23817

II"~'I '" ~'I ~I I ~'III"II'~

JAR'

The qUlIlity of the mntcl'i!lI.~ usea in tlw

manufacturc of this book is governell 1m
coni inllca postwar shcn'tagc8.
 McGRAW-HILL PUBLICATIONS IN
PHARMACY
A SERIES OF TEXTS A.ND REFlmENCE WORKS
OUTLINED BY 'rIlID FOLLOWING COMMITTEE
EV'VARD SIJEABl!J,C'lL(J.irman awl Oon8ldtino Editor,
Dean, Western Reserve UniverBity, School of Pharmacy.
JOHN O. BEAno" ,j
Profeasor of l'harmn,oy, University of North Carolina,
School of Pharmacy.
W. W. CIIART~ms,
Dimctor 01 the Dureillt of Educo.tiono.l Rc"oarch, Ohio
State Univcl"ait.y. Hcccntly Director in Chl"gc of the
Commonwealth Study of Phnrnlllcy.
H. C. CnnIS't'EN"I!lN,
Seoretary NILtional AB"Oci(,\ed DOllrd. of Pharmacy,
Chicago, Illinois.
ZAD.\. M. COOl'ElR,
ASBOcio.tJo Professor of Pharluncy St,n.to Ullivcrsity of
I

lOWI" College of Pharmacy.

ANDRBW O. DuMlOZ,
Dco.n, Univer.iLy of Mal'ylo.ll(l, School of Plmrmo.cy.
CIJAIR A. DYl!I,
Dcnn, Ohio State UnivcrfJity, College of PhlLfnU\cy.
C. W. JOHNSON,
De!ln, University of Washington, College of Phnrmncy.
D. D. H. JOHN"ON,
Dco,n, University of Oklaholnl1, School of Pharmacy.
C. B .ToRnAN,
Dcnn, I>ul'duo UUiVCfBitY1 School of Phnrnl[~cy,
TOWNJlJ~ R. J.,l :roII,
l

Do"n, University of Floridll, School of PluU'm"cy,

RUFUS A. LYMAN.
D~u.n, University of Nebrnalc 0. , College of Pho.rmncy.

WOItTLEY F. RUDTl,
Dean, Medicnl College of Virginia, School 01 Phnrm"cy.
CUAS. II. STOOKINCI,
Associnte ProfeRROl' of PlmrmltOy, Univeroity of Michi-
gall, College of l'hurIlll\cy.
An~o VIInlolnvER,
Professor of Biology "lid Phm'IIlncognoBY. Phnadelphi"
Collego of 1'IuU'm,,,,y.
A. L. I. WINNE,
Scerctu,l'Y I Virp;inin. Stato Board of Phu,nuncy. Ricll1nond.
Vil'ginio..
 McGRAW -HILL
PUBLICATIONS IN PHARMACY

B'ul'laoc, Ew'i, Leo and RiNiTL(J-

FUNDAMENTAL PRINClPLES AND PROCESSES OF
PHARMACY
Jenkins and DuM.2i-
QUAN'fITATlVE PHARMACEUTICAL CHEMISTRY
Jordan and D.Kay-
QUALITATIVE ANALYSIS FOR STUDENTS OF PHAR-
MACY AND MEDICINE
Lynn-
PHARMACEUTICAL THERAPEUTICS
Spease-
PHARMACEUTICAL MATHEMATICS
 QUANrl~ITArrIVE
PI-IARMACEUTICAL
CI-II~MIsrl~RY

CONTAINING TH.EORY AND. PRACTICE OF

QUANTITATIVE ANALYSIS Ar:PLIED
TO PHA!tMACY

BY
GLENN L. JENKINS, PH.D.
I'rof".,.qor of Pharrnn<,,,ulicrll Chcrn'istr1/, Collc(}c of Phmuuw1/,
U nivc'rsity of Jl{innesoliL

AND

ANDREW G. DuMEZ, PH.D.

Profl'8Nor of Plwl"1nnc'U and Dean of I.he 8chool of Ph""'lnac'U,
UnivlJ)'siiy (if Ma1'1Jlanrl

SE(!OND EDl'l'ION
rrNN'l'1.l 1~1 PHNHHI(J~

MuGRAW-HILL BOOK COMPANY, INc~

NEW YORK AND I,ONDON
1937
 COPYnIGH2', 1!):31, W:W, BY 2'HlD
Mc(lltAw-lIrr,r, BOOK COW.'ANY, INC.

All 1"'i(/ht8 reserved. 'l'his book, III'

11ar'is the1'eof, may not be I'cp'l'orlncctl
in any form wWwut 1Ie1'lIdswion of
the 1JllVlishc'l's.

THE USE IN THIS VOL,.UME OF CERTAIN PORTIONS OF' 'THE:

TEX.T of THE: UNITIZO STA.TES PHARMACOpOEIA IS BY VIRTUL:::
OF PERMlSsrON RE;CErV,EO FROM THE: BOARD OF' TRUST~~S

OF THE; UNITED STI\.n:s PHARMACOPOEIAL CONVENTION. THE:

SAID BOARD oF' TRUSTEES IS NO. fU;:sPON5IaLE FOR ANY
INACCURACY' ()F' qUOTATION Nott FOR ANY ERRORS IN THE
STATEMENT oF' QUAf'ITITIES OR PI!RCi!NTAGE STRENGTHS.

Pa:;.RMISS'ON 'to USE FOR COMMENT PARTS OF THE: TEXT OF'

THe: NATIONAl. F'OR,MUL.ARY, SIXTH J:;DI'I'ION, I~ THIS VOl..UME
HAS BEEN GRANTED BY THe: COMMITTEE ON PUBLICATIONS BY
THE:: AUTHORITY oF' THE COUNCIL. OF' THE: AME:RICI\N PHI\RMA...
CEUT'~"" ASSQCJATJON.
 PREFACI~ TO THE S]~COND EDITION
Since the puhlieation of the first edition of this book, numerous
challges hnve been made in tlw officinl methods of analysis of
phtll'maeeutical materinlfi in the United States Pharmacopoeia
Xl nnd in the National Formulary VI. The detailed procedures
and diseuRsiom; have been changed 01' rewritten in the present
edition to conform to the rnviHcd official methods.
The features that distinguish thil:! edition from the preceding
one are:
1. The contentH of the hook have be(;n rearranged into three
parts inste[tcl of four. Part I treats of general methods of gravi-
metric nnd volumetric analysis, Part II treats of physico-
ellO.mical methods, and Part III contains the Rpeeial methods of
pharml1cellticnJ analysis. In tho deletion of Part IV of the first
edition pertaining to lloll-ofiieial methods, the ehapters on
Hydrogon lOll Conecntmtiou, PllOtometl'ie Methods, and
Electrolytie MethodR have 1>oen placed in Part II under PhYRir,o-
dwrniealmethods. The elmptor on Ultimate Analysis has been
deleted to 8~we Rpaee and because a eompl'ohemlive treatment of
this suhjeet properly belongs in It course in organic tlul1lysis.
2. The theory has been brought up to date and many of the
ehapters lllLve been largely l'ewritten. Sections on Cn1eulations
aud ErrorH, Significant Figures, and Logarithms and Their Use
have 'beon added.
3. A lHlmber of llew exerciseR have been added and many of
the former exercises have been replaced by new ones so that the
instructor has a wielor range of selection.
4. The qnestions and problems have been extensively revised
und added to.
5. The method of presentation employed in the first edition,
wherein typical analyses are expla.ined simply, clearly, and
directly in considerable detltil, has been retained. Extensive
ta.bles classify all the official methods according to the type
methods which are described and explained.
vii
viii PREPACE '1'0 'l'IIJiJ 81iJCONlJ liJf)J'l'ION

The wide popularity that the first editioll or this hook ImH
enjoyed among students and teuehel'R hltH beml gratifyilltJ; to tho
publishers and to the nuthors. It is hoped that tho IWW (~dit;ioll
will similarly eommend itself to n Htilllnrger cin:Je. To tlw many
kind friends who have so gellorow-;ly ai(kd hy mnnllH of' ltoJpf'nl
suggestiolls, the l1ut]tOl'1'l (loHire to eXllJ'(iHi-: their thankR.
Gr,IDNN :L. ,h:NKINR.
ANIHUlW G. Du lV)I,lZ.
MINNJ%l'OLIH, MINN.
BAIJL'IMOIm, Mil.,
l"ebl'unry, 1937.
 PRKFACE TO THE FIRST EDITION
The objeet of this hook iR twofold: First, to furnish stndents of
pharmacy with [L systematie course eovcring all of the qunntita-
tive chemical and physical methods official in the United States
Pharmacopoeia amI the Nn,tional Formulary through the Relec-
tion and explanation of typical procedureR. Second, to present
SOllle of the generally applieable, non-official methods of analysis
whieh are widely used in pharmacy and with which all students
pnrsuiug the profm;Hiol1 should be familiar. In both instances,
the theory and practice of analytieal chemistry as applied in
qmtntitative phtumaceutical procedures have been correlated.
The use of the book ns a text should be supplcmented by lec-
ture and recitation instnwtion. It iH obviously impracticable to
ineludn detailed explanationH of all of the quantitative determi-
nations in the Pharmacopoeia, and National Formulary. Typical
analyses illustrating all of the various methods have therefore
beon Relected and explained in eOllRiderable det.ail, while those
determinations requiring the same or closely similar procedures
are indicated in tables 01' othcrwhm. The instructor may select
other determination" from the Pharmaeopoeia or National
FOl'Jlllllal'Y and nssign to the students the task of applying the
theory and explana,tion of the procedure involved aH given ill the
text.
The book is divided into four parts. Part I is intended for
use with HtudentH who have completed a full year of work in
general inorganic chemistry and qualitative analysis. Parts
II and III preferably should be given after the st.udents have
completed inorg!1nic and organic chemiHtry. Part IV should be
given to advaneed students who have acquired a thorough knowl-
edge of qnantitative technique. In the authors' classes, Part I
ill givcn during the second half of the second year in a course
eovering sixteen weeks with one four-hour laboratory period
each week. In t.his course, about twenty assays are performed
ix
x PRB}I'A(}JjJ '1'0 '/'lIB FIRS']' HDJ'l'f{)N

in addition to the exereis()s pertninill!!: to Ow IUlalytienl halmH:n

and stnnd!trd solutiolls. Parts II anci III ILI'n givnll ill the fHl(:OIHI
half of tho third year in it nolU'SO of instrur:tioll r:oVel'illg; sixtooll
weeks with two three-hour Jahomtory pnriocis pm' w('C'!c
Approximntely thirty-two eXnrr:iS(:H aro pnrfOl'llWd till rill!!: this
eourse other than thoso whieh deal with the: pr()]mra.tioll of stnnd-
ard AolutiollS, cte. Prar:tienIIy nIl of' Part IV iH LaknH up alollg
with other work ill a eourse in foocl and drug analysis givcn! ill
the fourth yeul'. The Humber of exoreiSOH that eall 1>0 (:ov(!n:d
in any given eourse will depend Oll tho prepamtioll of tho At1l(1mrt:-:,
the amollnt of time devoted to the suhjeet, and the fnr:ilitinH
n vailable. The large number of exereiflCS iJ1(~lnd()d providn for It
considerable dngree of elasticity in the time devol,od to the HLndy
of qua,ntitl1tivc methodr; and permit the instructor to Vt~ry tho
exereiscH with different classes.
The authors wish to acknowledge their appl'eoiation of tho
eourtesy extonded by the following eompanim; which granted
permission to usc illustratiolls from their (:atuloguos and hooks:
A. H. Thomas Company; Bausch ulld Lomh Opticnl Company;
Central Scientific CompfLllY; Christian Beeker, Ineorpomt(!d;
E. H. Sargent [mel Company; LaMotte Chemical ProdlLc:ts
Company; and Leeds & Northrup Company.
Notiees of error and suggm;tions for thc: improvement; 0)' tlw
text will be greatly appreciated by the authors.
GLENN L ..hJNKINS.
ANDm~w G. DuMBz.
BAL'rIMORlil, MD.
December, 1930.
 CONTENTS
PAOEl
PREFACE TO 'I'HE SI~COND EDI'l'ION. vii

PUEl'ACE TO 'l'lm ]i'IHS'l' BDI'l'ION ix

IN'l'RODtTC'l'ION . , . . . . . . xix
Definitions and scopc of qUlLlltitl1tivc phal'trutccnticn] ehcmistry-
Refcr()llec~.

PART I
General Methods Used in Official Pharmaceutical Analyses
CHAPTER I
REMARKS ANIl GENEUAL DlREC"l'IONS . . . . . . . , . . . . . .. a
Snmpling-Calr:uhtion of results !lnd fWI'Ol'S-GCJl(lJ'[Ll opel'ntiolts-
The :LIudytienl Imlallce-Weight.s.

CHAPTER II
GUAVIME'l'RIC ANALYSIR . . . . . . . . . . . . . . . . . 29
'fhcory of ionilmtioll-Heversible renetions-Soluhility produet,
prillciplc--Colllmoll ion effect.

CHAPTEH III
GUAVIME'l'RIC ME'rHoDs . . . . . . . . . . . . . . . . . . . . . 37
A::;sny of sodium chl()rid(~, of sodium sulfate, of mercuric chloride, of
r:nlcillm gly(icrophoRphllt.(~, of alum, nne! of solution of magnesium
citrtttc.
CHAPTEH IV
PmNCIl'LES 01" VOLUME'I'RW (TI'rIUMETRIC) ANATJYSIS. . . . . . . 57
Dctillitionf)---VoIIIUlct,l'ic Itpparnt\l~-The calibration of volumctIj[)
npp:tmLus--NrmtmlizlLtion methods: Theory-Illdicntors-
St.andard Holut.iollS-Prcp!Ll'lltion and standardization of normal
hy(lroehlol'i[l :.wi(l, of normal sodium hydroxide, and of teuth-
normal hariulll hydroxide.
CHAPTER V
Ar,IlALIMETHY . . . . . . . . . . . . . . . . . . . . . . . . . 86
Direct titration methods: Asstty of sodium bicarbonate, of sodium
hydroxide, and of sodium salicylate.
xi
xii CONTEN'l'8
PAnl!
Residunl titration methods: Assay of zinc: oxide, of P()tll~sillm and
HOI[ium t!trtratc, of m!lgnmiia nmgmll, of metlwnnmiue, of tloll1-
Lion of Itmmonium !LCetate, anll. ()st.imatiull of lIitroglm by tho
Kjeldahlmothod,

CHAPTER VI
ACIIHME'l'RY , . . . , , , , . . . . , , ' , , . , . , . . , , , 101\
Direct titration mctliOlls: Assay of (li1uted HltlfllJ'i(~ al~id, of l\Ori(~
add, of tllblet.s nf sodium sll1ieybt.e, llTld of tlll'LI\I'i(~ IWitl.
Residual titratiolllllethods: AssflY of nroIDlltiu sulfuril\ add ailli of
tablets of IweLylslllieylit: twirl,

CHAPTER VII
PUECIPITA'l'lON ME'l'HODS , lUi
DeterminatiolJ of the end point.-ImlielLt.ol's, SttUl(bnl Holllt,jOIlH.
Preparation and stlllldardizaLion of telltli-lIol'lllltl ~;jlvet' lIitrate allli
of tenth-Ilormal Illtlmoniulll thioeyalllLi.(l,
Direct titration l11eth()d~: AHI-my of strollg Hilver protein,
Residua] Li tl'ation methods: ABilIty of Hodiulll ehlol'ido, of :lllJlllO-
lliulll bromide, of syrup of hydl'iot1ie fleid, lind of dixir of 1.lll'lllJ
bromides.

CHAPTER VIII
OXIDA'l'ION-REDUC'l'lON Ml~'l'HODS , , , , . , . 120
Theory--Standal'd solutiOIlA-Pl'epal'lltiou !Iud stlllldanlir.a1.ion of
tenth-normnJ potl1ssiuHl Pm'lIll111gH.lll1tc,
Direct titration methods: Assay uf ferrous sulfate, or 1'0(1\1(:1'11
iron, !Lud of solution of hydrogen peroxide.
Indirect titration methods: AssflY of oldeillItl gitlOollllto,
Residual tHratioll methods: Preplll'llt.jon mal Hj,all(lllrdizat;ioll of
tenth-normal oXfllic acid~AHsay of sodiulLl nitrite, of 111'0-
eipitated caleium carbonate, and of potassium (',hlol'ld;c.
Diehromate methods: Proparation of tenth-llOl'1nal jJOLltHl:iiultl
dichronl!\tc~Assay of lllass of ferroliH nll<l'bOlltLt.ll.

CHAPTEH. IX
OXIDATION AND REDUC'l'ION-IODOllilIil'l'IUC MBTHOI\S. . , , , . . , , Uifi
Starch indicator solutions, StancIllI'd SOlll LiollA: Pr()jlll!atioll mnl
standardization of tenth-normal sodium thiosulfate Holutioll
and of tenth-normal iodine,
Direct titmtion with standard iodine solut.ioll: AHsay of tll'H(mie
trioxide,
Dired titl'!1tioll with sodium thimmlf!lte: Assny of !:ollljlouml
solution of iodine,
Residual titration with standard sodium thiosulfate: AHsay of
mercurous chloride,
 CON'l'EN'l'S xiii
PAGE
Tit,rntioll of t.he iodine lih8raj,ed from pot.IlHsium iodide wit.h sodium
LhioHulfl1te: Assay of solutioIl of ferric chloride, of chlorinated
linw, of cnprie Sllifato, of sodium lLrSflllllte, of thyroid, and of
spirit of ethyl nit.ritn.
Titmtioll with tcnth-llOrl1l1tl hromine: Preparat;ion and st.a.ndardi-
zILtion of tenth-normal bromine-Assay of phenol ltnd of
ammonium hypophosphitn.
Titmtions with Atandn.rrl pota,ssinm iodate: Prepamtioll of flta,ndarrl
potnssiultl iodlde s()lut.ion--A~say of potassiulIl iodide.

CHAPTER X
GASOME'L'Hrc ME'I'HODS . . . 188
'l'hnory-AllPlLr:Ltus-Test of t.he llitronwter-Ass:LY of [~lLrboll
dioxido and of spirit of othylnitrite,

PART II
Physical Methods Used in Official Pharmaceutical Analyses

CHAPTER XI
SOLUJlII,rry. . . . . . . . . 208
Definit.iolls.
Determinnt.ion of the solnhilit.y of borie add in witter at 25C.

CHAPTER XII
SI'Jj;Glvrc GlIAVrl'Y AND DNNI-iITY . . . . . . . . . . 207
Met.hods used t.o determine the spceific grn.vityof liquids: The !lEW
of Jlyenomoi;cl's-DeterrllilllLtioll of t.he alcohol eontont of all
ollieinl preparation. The usc of the Wcst.pluLl bf1IaIl(:c-Detel'-
winatioll of the ;;pneifie gmvity of I), volatile oil. The use of
hy(\rolllot,ors.
Mct.hodH used to detormine tlw spoc:ifie gravity of solids: By weigh-
ing in watorDntorrninat.ioIl of the specific gravity of nmnphor:
By t'\w flot:Ltion ll1et,hoti--Detorrninatioll of the speeific grn.vity
of yellow wax.

CHAPTER XIII
MEIfrING, CON(lEALlNG, AND BOlUNG POlN'rs. . . . . . . . . . . . 226
Melting poin1;: Determination of the Illelting point of sll.lieylie Iwid.
Congealing point: Method of detcl'lninillg-DcterminntiOll of the
soliclifioatioll t.empemtlll'c of the fatty acids of eottonseed oil.
Boiling and distilling point: Deterlllination of the boiling point of
Cl1rbOIl tetrachloride. .
XIV CON l'EN 'l'S

CHAPTER XIV
REli'HACi'OMEl'l'HIC MElASnUElMEN'l'A . . . . . . , , . . . . . . , 24(1
Refractive index: Refmotometerl:l-Tho Abbe refl'llc:toll1otor,
DetermiuILtioll of the refl'llotiv(] index of oil of ol'llllge.

CHAPTER XV
Ro'rATORY POWER . . . . . . . . . , , 247
Definitions-Poitwi Il\()tcrl:l.
Determination of the sped fie rottttiotl of ;\111\1'08(\.

CHAPTEn. XVI
VISCOSITY MEM\UHElIl1J!lN'l'H. . , . . . . . . . . . . . . . . , 257
Dofilljti()ns~App[\ratul5: The Saybolt viseDsimn\.('r.
Determination of tho kinemati,\ viscosit.y of liqllid J!ot,roln(;um.

CHAPTER XVII
PlIO'l'OM1~'l'ltlC MI!:TlIOPS 011 ANALYSIS. . . . . , . . . . . . . . . 2l\O
Colorimetry: Det,cl'l'niIU1tion of tho ammonite ('.onten\. of Witter
Dotermiun,t,ioll of t.ilO amollnt of epinepill'ilW hyllr()l.hlol'hlo ill
solution of epinephrine hydrouhloride-Assny of (il'f)(\UH for ,~ol()r.
Nephelometry: Dnterminat.ion of the [tmount, of arselli,\ tr.ioxido ill
solution of arsonous lwi<i-Dctol'miulltioll of the n,mount of oil of
peppermint in spirit of peppermint and limit test for (:lllol'ilie and
sulfate in cnlr:iulll glucormte,

CHAPTER XVIII
DETERMlNA.'l'ION OF HYDROGEN ION CONCEN'l'ltATlON 272
Acid base equilibrium and pH.
Potcntiometrie methods: The hydrogen nlcetrolie'"-PlatinlzlLtion
of the hydrogen elcetl'odc--The calomel eioctro(k-Tho HdwlIlll
of assembly for hydrogen ion )1lC'thociH.
Determinatioll of the end point. of titration of hyclroohlorie add
with sodium hydroxide pot(mtiollletl'ieally-Nol,()s nnd 1)1'(:-
cautions-Determination of the end point of titrat,ioIl or twdio
acid with sodium hydroxide potel1'Uolllfltl'ieally. Dntol'minnLion
of the pH of satumtml boric acid solution, n,nrl of phYHiolop;ienl
salt solution.
The quillhydrone clcetrocle: Preparation-Usc of.
Determiuation of the pH of elixir of iron, quinine and stl'yei>nill(1,
elixir of pepsin, and tillctme of aconite hy meallS of the quin.
hycll'One elee f;rodll. .
The glass electrode.
Colorimetric methods: Indit:l1tol's-Bl1fflJl' fl()llttioll~-Colol' Htltud-
artis-Color comparators.
Det(\l'!llinatioll of the pHof solution of epinephrine hydl'lwhlol'i,h:,
and syrup of hydriodic acid,
 CON1'EN'l'S XV
PAOlO
CHAPTER XIX
. 305
ElnoLrilml units :tnd fundamental 1[L1V~~Thcory--AppamtuB.
Assay of !'opper Hulfate lind of mercuric: chloride. Other electroly-
tic a8Sa.ys.

PART III
Special Methods Used in Official Pharmaceutical Analyses
CHAPTER XX
ASH AND Mors'I'TJIm DE'l':mUMINA'.l'IONfi.. ......... 323
Ash content: Determination of the total and tMid-insoluble ash
(lout.cut of digitalis leaf.
Muisture content: DeterminatioIl of t.he moisturn content of l10llcia
-Det,nrminatioll of the moistnre (lontont of digitalis leaf by the
toluene distilltttioll md,hod.
CHAPTER XXI
EX'l'HAC'I'IVID AND CRUDE li'mEH CON'rEN'!' . . , 33tl
Volatile and !loll-volatile ether-soluble extl'l1(:tive: Dotermination
of t.he volatile ane] nOll-volatile ether-soluble extl'l1ctive of dove.
Aloohol-solublc extractive: Assay of benzoin.
Wa.ter-HOluhle ext.raetive: Assuy of ltloc.
Purifie(l petrolUUIll henzin DxtmetivD.
Crudo fiber: DuterminM,ion of the crude fiber content of nioves.
CHAPTER XXII
CONSTANTS 0J0' FATS, FA.'l"l'Y OILS, WAXFlS, BALSA.MS, RESINS, ETC, . . 345
Acid number: DeLermination of the acid yalue of rosin.
Snpouifieation v{due: Dcterminntion of tho saponification value of
(mt [;ollseecl oil.
}jjstnr number,
Un~lLpollj(iable
matter.
Iodine value: Determination of the iodino value of olive oil.
CHAPTER XXIII
ASSAY OF VOLA'l'U.J; OILS . . . . , . . . . . . . . . . . . . . . 361
Methofls of p;elllJralltpplielltinn: Sppeifio grl1vity--Rotatory power
-Hefractivll illlkx-Uollgellling; point-Distilling point-Fl'lIc-
tional clistillntion-Suluhilit.y.
AHslLY for ester COl1tl!Jlt: Pwpamt.ion of half-n01'lIlnl aleoholie potas-
SilUll hydroxide-Assay of oil of peppermint fo!' totul COlters.
AS~l!LY for aleohol. eOlltent: As~ay of oil of poppermint for total
IIllmthol.
As!-my for aldehyde I.'.olltont: A~~!tY of oil of bitter !Llmond for
bcnzaldehyde content.
XVl CON'l'IiJNTS

Assay for ketone content: Assay of oil of camway.

Assay for ]lhenol content: Assny of oil of dove.
AS8fty for hydrocyanic acid content: Assny of oil of bitter nhnol\c]
for hydroc:yanie add.
As;;ay for ase:llridol content: Assay of oil of eheIlO]l()eliulll.
Assay for nllyl isothiocyltllltte: Assay of oil of lllll:>tanl.
Assay for volatile oil ill spirits: Assay of spirit of pep]lol'lllillt.

CHAPTER XXIV
ALKALOIDAL ASSAYING . .
General principles: Sourees of el'ror--Thoory of 1[i:;I,rihut.ioll
cocffieiellt-Choioo of inelicatol':;~--T()st solutiollH.
General procedurlls: Seleetion of tho HILlIlpic:--ExLr:u:Lioll wiih
immiseihle Holvonts----EvlLjlOl':ttioll of org:wie: solvent:;-----(}ravi-
metrie rleterminlttion of alkaloid~-Vollllllctrie t1eterllliwtLioll of
ttllmloirls.

CHAPTgR XXV
OF~~ICIAI, TYPE lVIE'l'HODS .
Alkaloidal assays hy aliquot, parI, method.
Gcneraillroceduro: Extl'l1ctioll of the drug--nooILlIting Uw niiquoL
portion-SIHtking out. with aeid-Sh:tldng out. wit.h illlllliH(:ibln
solvent-Determinntioll of the alkaloidal cont{mt.
Gravimetric assltys: Assay of hydrastis for ci.lwl'-solnhlo nllm-
loills, of cinchona for total :dlmloirls, ami of eompollnd t.inet.uro of
cinchona.
Volumetric ItSSI1YS: Assay of ipecac for ether-soluble allmloiehl ILnd
aSIli1Y of i1reca.
Allmloidal assays hy thCl totnl oxtraetiOIl methofl: Assay of hyo-
scyamus leaveR.
Assay of preparations of hyoseYlunus, belladollnll, ILnd stmlllOllillll\:
Assay of tincture of bolladonua and fluidextraot of belladolllllL
leaf.

CHAPTER XXVI
ALKALOIDAL ASSAYS BY SPECIAL lVIE'rIIODS . . . IHS
Assay of opiuIll.
Table of offieial substances assnyecl by the s!tme method as opium.
Assay of eolehieum.
Table of substances nSHayed by the amno mothod liS colohieum.
Assay of nux vomioa.
Assay of caffeine oontailling drugs: Assay of guarana.
Table of of1icial dl'l1gs and prcpamtions assayed for (lILffoinc,
Assay of alkaloidal SllltS: Assay of citrated ()~tffeine, of {lur:aiwl
hydrochloride, of t.heobromine wit.h sodium sali(lylate, and of
theophylline with sodium acetate.
 CONTENTS xvii
PAGEl
CHAPTER XXVII
(hUEU OI'l'lCIAI, Af\i'\AYS INVOlRINO 'l'lU: UB}] 01" IMMISClllLI~ SOLVENTS 437
ASHILY of (~allthlLl'ideH--ARH!1Y of ltspidium--Assay of j!Llap-Assay
01 t.ahletH of phcno\Hll'llital.

CHAP'l'ER XXVIII
AHI:\A Y (W ENZYMJH)ON'I'AINING SURH'l'ANCES . . . . . . . . . . 44,1
Assay of Ill)(lHill------AslmY of panCI'Oatjll fo!' stlLreh digest-ivo POWOl'--
Assay or IllLllo)'(mLin for clLsein digest.ive powol'--AssLLY of l'flllllili.

TAHl,g ()lo' LOilAltl'I'UW-l AND AN'l'll,OGAHl'l'HMS. 453

INllBX . . . . . . 457

T"m,l~ 01" A-l'O!.UC WI~Hm'l'S _ . . . . . _ . . . _ . Inside hack e.over

 INTRODUCTION
Quantitalivp- phm'maccutical analysis may be defined as the
applientioll of t.he procedures of quantitative analytical chemistry
'to the analysis of the materials used in pharmacy and, especially,
to the determination of the purity and quality of the drugs and
chemicals offieial in the United States Pharmacopoeia and in the
National Formulary. A complete chemical analysis of any
substance requires the establishment of the iUClltity of its
compoll(mt pnrts by qualitative analysis and the determination
of the proportioll ill which tlwse components are present by the
processes of quantitative annlysis.
Quantitative chemiea,l methods are commonly classified as
organio and inorganic; gravimetric, volumetric, gasometric, and
eleetrometric. It is impractical, however, to acUlere strictly
to thiH cllWHifi(~fI,tion in prps(mting the subject matter of quantita-
tive plmrmnceutical analysis since certain phases of the work, as
alkaloiclttl assaying and the assay of essential oils, have been
developed to a point where t.hey constitute a distinct subject
matter in which the prineiples involved and the technique
required are the SlLme whether the procedure be gravimetric or
volumetric.
Quantitative pharmaceutical analysis may be appropriately
subdivided aeeording to various procedures each of which requires
It special technique, as follows:
Gravimetric analysis, or the separation, by extraction, precip-
itation, or other means of the constituent to be determined
either in the natural state, or in the form of a definite compound
the eomposition of which is known to the analyst, and weighing
the resulting product.
Volumeb'ic analysis, or the determination of the volume of a
solution of known concentration required to react with a given
amount of the substance to be analyzed.
Ga.somctr'ic analysis, or the measurement of the volume
of a liberated gas or the decrease ill volume of a mixture of
xix
xx [N'l'RODUC1'ION

gallCR when a suitable reagent is Hseel to remove one of the

gases present.
Physico-chemicalmcthods of anal1}sis, or those which al'e based
on the utilization of Rome specific physical or ehemieal property
or properties of the su!mtallcc in HR qU!1ntitative o:;timl1tion.
Those physico-chemical methodR eommonly applied in qlUtntitt1-
tive plmrmaceutienl c:lwmiRtry are optomotric, rcjmctomctr-ic,
elect1'ometl'ic, and colorimetric. They comprise some of tho
most. valuable methods used in ttualyticltl proeedure. Sinee
eaeh of them require special treatment, they will be discussed
separately.
Sl!ecial. methods, or those whieh requiro a distinct type of
technique, as alkaloidal assaying, require separate treatment ill
order to preserve unity 01 sUbj(lct matter.
The official assay methods serve as an exact measure of tlw
purity of a substance only when the results are considol'(lIl ill
conjunction with the qualitative tests. Thus, in the assay of
zinc oxide, thc purity of the oxide as determined by as:my must
follow qualittLtive tests for other metals which if present would
be estimated as zinc oxide. A eomplete analYHis, therefore,
l'equires the qua,littttive identification of nJI substances prOHont
as well as their quantitative f~stimation.
The theoretical eonsidemtions applicable in any given analYRiH
are inherently a part of the analytical procedure. They should
be mastered hefore a det,ermin:1tion is made and carefully appliu(l
throughout each step of the procedure.
Those who desire a more comprehensive treatment of spcni:tl
aspects of analytical proceduro will find the following texti:\ lmd
reference books of value:

Textbooks
1. BASS lilT, "The Theory or QUltntitative Analysis," Alfred A. Knopf, Ine.,
New York, 1925.
2. BLASDALE, "Principles of Qll!Llltitlttivc Analysis," 3d cd., D. Van
Nostmnd Company, Inc., New York, 1928.
3. CLO'YES and COLEMAN, "Quant.itative Chemical Analysi8," 13th e(l.,
P. BlLtkiston's Son & Company, Philadelphia, 19a1.
4. CUMMINS and KAY, II A Textbook of Quantitative Chemical Analysis,"
6th ed., Gurney and Jackson, London, 1934.
5. ENGELDlilR, "Elementary Quantitative Analysis," 2d ed., .Tohn Wiley
& Sons, Inc., New York, 1936.
 I Nl'lWDUCT ION XXI

6. F AI,ES, "Inol'gfmic Quail tittL tivc AuuJysis," D. Appleton-Cen tury

CompfLIlY, Inc:., New York, 1925.
7. Ji'AltN"SWOH'l'H, "Theory and Technique of Quantitative AnalY~lH,"
,John Wiley & SOIlS, Inc., New York, 1928.
8. li'AY, " An AdvttllCed Course in QUltlltitl1tivc AlllLlysis," 2d ccl., ,John
Wiley & SOilS, Inc., Now York, 1922.
9. GUY tmli SKImN, "A Course ill QlUtntittttive Analysis," Giml 11,1ll\
Company, Boston, 1932.
10. HAl,r" "A 'l'extbook of Qmmtitativo Annlysis," 211 ed., John Wiley &
SOliS, Inc., New York, 1935.
11. HICNIlNL, "QlULntiLntive Antilysis," Ginll :Lud Company, Bostou, 1921).
12. I';:orlrIlOl'F lind SANUJuLr" "Textbook of QULLnt;itntive lnorgnllie Arudy-
sis," The MaemillaTl Company, New York, 1936.
18. MAllIN, "QUfLIltibttivlJ Analysis," ,!th eel., McGraw-Hill Book Com-
pany, Ine., Ncw York, 1932.
H. Or,KIm, "Quuntitative Amtlysis," lith (,(l., D. Van No~tl'/Ind Company,
IIlC., New York, 1919.
11). Popor'l', "Quantitat.ive Analysis," 3r1 m!., P. Blakist.on's Son & Com-
pallY, Philadelphia, 1935.
16. RIEMAN and NEUSS, "Theorctical Approach to Quant,itativc Analysis,"
McGmw-Hill Book Company, Inc., New York, 1037.
17. SCIIlMPF, "Esscntials of Volumetrie AlIalysiH," 4th cd., John Wil(~y &
Sons, Inc., New York, Hl2H.
lB. S(!O'l"l', "Esscntittls of QUllntit,!Ltive ChcmiCttl Analysis," 2d ed., Cltemi-
eal PlIbli~hillg Company, EastoIl, Pa., 1981.
1Il. SMI'l'H, "quantittttive Chelni('.[ll Ailitlysis fur Beginning Students,"
3d cd., '1'he Macmillan Company, New York, 10:33.
20. S'l'OCK and S'l'Xm;EU, "Qultutitl1tive Ch()lllie~tl AnalY8is," MeGmw-
Hill Book Company, Inc., New York, 1935.
21. TALHO'l', "Introductory COUl'Illl in Quantit,al;ivc Analysis," 71,h ccL, The
MaemilJan Company, New York, 1931.
22. WILr,AUJl and FURMAN, "Elcmontary Quantitative Analysis," 2d cd.,
D. Van Nostrand Company, Inc., New York, 1936.

Calculations
1. I:-I.<l.lIULroN n.nrl SIMPSON, "Calculations of Quantitative Amllysis,"
2d ed., McGraw-Hill Book Comp:LIlY, Inc., New York, 1927.
2. LONG and ANDEUSON, "Chemical Caleubtions," 3d ecl., McGI'n.w-Hill
Book Company, Inc., New York, 1936.
3. MEr,Lon, "Higher Mathcrn!LLics for Students of Chemistry and
Physics," 4th cd., Longmans, Green & Company, New York, 1922.
4. MILl,NU, "Calculations of Analytical Chemistry," 3d ed., McGmw-
Hill Book Company, Inc., 1921.
5. MOORE, "Logmithmic Reduction T!Lbles, for Student.') of An!tlytical
Chemistry," Ginn and Company, BostOIl, 1913.
xxii IN'l'JWDUC'l'10N

n. WILKINSON,"Calculatiolls ill qu:mtita(.ivp AllalysiH," MeGraw-Hill

Book Company, Inc., New York, 1!l28.

General References
1. ALLEN, "Commel'eial Organic AlUtly~iH," lith ecL, P. Blakisi;oll'H Hon &
Company, Philadelphia, 102H.
2. GOOCH, "Methods in Chemical Aultly::;i::;," ,]olm Wiley & HOliK, Ill!'.,
New York, 11ll2.
3. GRII'I"IN, "Teelmical Metho!ls of AnalYHiH," MeGraw-Hill Book (\J1n-
pltllY, Inn., New York, 1021.
1. KOVl'IIOl'~' and PUR MAN, "Volllmetric: Analysix," .John WilClY & !:lOll;!,
In!:., New York, 1920.
5. MEr,r,oR, "A TrcatiHo on Qllan(,ji;ntivD Tllol'ga.llie AllalYHi~," Chal'lnH
GriHin & COlllpany, Ltd., Loudon, 191:3.
(i. ScoT'r, "Stl1l1dlll'd Mot,hods of Chcmic:al AnalysiH," 41,h niL, D. Vall
Nostntncl COmjHLllY, Tile., Nnw York, 1921).
7, SMI~'H, "AJllI1y('ieall'l'oeeSHcs," gdwH.l'cl Arnold HlId Co., LOlHlolI, 1!l2!1.
8. Su'r'l'ON, "VolUlllotrie AnalysiH," 12('h ed., P. Bln,kiHtoll'H HOll &, Com-
pany, Philadelphia, 19:35.
9. 'fUEAIlW)'lLI, and HALT" "QuantitatiVf1 Al1IdYNis," 8th mI., John Wiloy
&; SOilS, Inc., Now York, 1\)8 Ii.
10, VII,r,AvEccIIIA, "Applied AIlIl!ytic:nl Clwmi:;;('l''y,'' P. BltddHt.OIl'~ H,ou &
Company, Philadelphia, 1918,

Drug Analysis
1. DUAGENDOIl.FF, "Plant Analysis, QUll.litativo and QUt\ll Litati Vll,"
. B:illiel'e, Tindall !tnd Cox, London, 1884.
2. EVErtS and ELSDON, "Analysis of Drugl:! and Clwmiollls," ChILI'kH
Griffin and Company, Loudon, 102n.
3. ]i'UI,r,ER, "The Chemistry [mll Analysis of Drugs awl Medieilll:H," .Tolm
Wiley & SOIls, Inll., Now York, 1920.
4. LYONS, "Practical Standardization of D!'ugs," Noison &; Co., ])u(.l'oi1,
1920.
5, "National Formulitry VI," 6th eeL, Maok Printiug CCllllPIlIIY, Ellston,
Pa., 1936.
B. NELSON, "Introduction to the Analysi~ of Drugs ami Medic:ineH,'! .Tohn
Wiley & Sons,.Inc., New York, 1910.
7, "Now and Non-ofilcial Remedies" ("N.N,R"), AIll!lric'lIll MedLenl
Association, Chicago, 1936.
8, "Pharmacopoeia of tho United States Xl," 11th (,llV., Made Print.ing
Comp!IIlYI Easton, Pa., 1936.

Food Analysis
1. BLn'H, "'Foods, Theil' Composition und . Analysis" .6th mI., D. VIm
Nostrand Company, Inci., New Yoi'k, 19(1).
 I N'l'RODUC'l'ION xxiii

2, BRIDGES, "PoOlI and BevorfLg;o AnlLlysi~," Lea Feheger, Philadelphin.,

1935.
3. Cox, "Chemical Antilysis of Foods," p, Blalciston's SOIl &; Company,
Philadelphia; 1921\.
4. LgACH and \VINTON, "Food Inspection [wd Analysis," 4th ed" .Tohn
Wiley &; Rom!., Jlle" New York, HJ20,
5. LEI"I'MAN [mrl BE.ur, "Food AnILlysis," P. Blaldston's SOil & Company,
Philadelphiu, 1\)05,
(i. SHERMAN, "Ol'glmie AnalYHiH," The M:u:millan CompfLllY, New York,
1!)20.
7. WOODMAN, "Food Analysis," 3rl eel., MoGraw-Hill Book Company,
IIlC" Nnw York, I93l.

Food and Drug Analysis

1. LIVJmRIDJ~(iB, "Adulteration and AnlllysiH of Foods aml Drugs," D.
Van Nostmnd Company, Ill!)" Now York, 1932,
2. M aOH and P Am'HlDGEl, "Aids to the Analysis of Food nUll Drugs,"
Gt.h mI., William Wood and Compltny, Baltimol'n, H)35,
3. "Methods of Analysis of the Associatioll of Offidal Agricult.ul'nl Chc!l1-
illts" (A,O.A,C.), 3d od., Assoc, Off, Agr. Chmn., Washillgtoll, D. C.,
1932.
4. PA HHY, "The AultIYHiH of Food and Drng;s, Chemieal and Mir:l'mwopicaJ,"
81:0tt., Grecnwood & Sl)})!>, Lonrlon, 1911.
5, TnUUK'l'ON, "Phal'mar:eutir:al amI Food Analysis," D. Vau Nostrand
Company, Inc" New York, 1922,

Special Analytical Methods '

1. BLAIH, "The Chmnieal Annlysis of Iron," 8th ccl., .T. B. Lippineott.
Compnny, Philurlelphia, H)18,
2. Ihu'I"l'ON, "ConrIuetillletric AIlIllysis," D. Vall Nostrand Company,
Inc" New York, 1934,
3. - - - , "Hydl'og;OIl Ions. Their Determination lint! Irnportunoe in
Pure find Inclulltrial CllOlllistry," D, Van Nostrmld Company, Ille.,
New York, 1932,
4. CLiI UK, "Deterlnillatioll of Hydrogoll IOlls," 3d ed" Williams & Wilkins
COl1lPIIllY, B!tIUlrlore, 1D28.
5. DENNI8, "GltS AnalysiR," Tho Mamllillan Company, New York, 1918.
(I. DUl'rJ!JnICH, "Analysis of ROBins," Seott, Greenwood &; SOIlfl, London,
1020.
7, Gn,IlEMEIS'.l'BH, H(WI"MAN, aIHl KnBMI~ltA, "The Volntilc DiIH," John
Wiley &; Sons, Inc" Now York, 1922.
8. HAI,nANN ami GHAHAM; "Mel,hods of Air Analysit:l," 4th ocl., CIH~rles
Grimil & Company, Ltd" LondoIl, 1935,
9. HJWJ~i-lY, "Chmllieni Aualysis by X Rays lind Its Application," Cornell
University Pross, Ithac:a, N, y" 1982,
xxiv 1N'l'[WDUC'/'[ON

10. HILl,IDDnAND and ]~UNDIDLL, "Applied Illorll;tlllie AualYI;is with Hpedal

Refel'enee t,o the ArmlysiH of Metals, Minernb:, lind HookH," .Tolm
Wiley & SOllS, Inc., New York, 1\)20. .
11. .KOIil'HOFI", "Tho Colorimetrie :tnt] POj,()!ILi(}]nd,ritl Dl\j,(1l'111in:\,Lioll o[
pH," John Wiley & Sons, Inc., New York, 1931.
12. - - - , II Conduetilllotrie Titmt;iollfl," T. S1.einkol)l'f, :LlliplIig, ID2::1.
13. - - - , :md Ji'UHMAN, "Potelli,iOluei;rie TitrnLiolls," 2d ed., .rOllil
Wiley & SOIlS, 11111., New York, ID'll.
14. LACEY, "Illstrumentnl Methods of Chmuieal Analysis," Tho MIL('lllilllLll
Comp:Lny, Nnw York, 102'1.
15. LmVKOWI'I'HCH, "The Chcmieal 'l'eohllOlo!2;y itllrl AllalYHis of (lilli, Fat:;
amI Waxes," Oth ee!., 'fhe M:wmillnll COlllpa.ny, New York, U)2:l.
Hi. LUNGE, "Teelmie:Ll GRS AnalYHiH," n. Van NnH(,mlHI Cc.lHlpH.ny, 1m'.. ,
New York, 1934.
17. IV[AHIN ami CAlm, "QtlftuLitativp. AgrilmlLnl'lll AllalysiH," MIlGmw-IIil\
Book Company, Ino., Now York, l!l2:l.
18. PIll'rmwl and VAN SLYKlil, "QllantitaLivp. Clillielll CltmlliHj,I'Y," WilliamH
& Wilkins Company, Baltimoro, 10:32.
10. PUlil(lL, "QuantitrLi;ivp. Orgnnie Mieroll.llalysis," 2d EIlp;li;;h {I(!., P.
Bbkiston's Son & CompfLUY, Philndu]phia, lOan.
20. RAIJLI<JY and GHAW!', "Fluorosemwo Analysis ill Ult,ravio](Ij; Lip;hl.,"
D. VIm Nostrand Company, rIte., New York, 19:3:3.
21. RAE fLnd REILLY, "Physico Chernieal Pmetienl Ex()rdBll~," M(I!'.lllUIll
aud Compltny, Ltd., London, 19:15.
22. S~1l1'H, "Electro-analysis," 6th od., P. Blttkistnn's Ron & Company,
Philadelphia, 1918.
23. and SNEI,L, "Colorimetric ]\IIuthorls of Allnl,VsiR, Tn(~lud inl~ HOJ]w
SNJiJI,T,
Turhidimetric llllel Nepholometrie Mothods," D. Van Nostl'lLlJI!
Company, rne., New York, 1936.
24. WINKLER, "Ha.rulbook of Teehnienl Gas Analysis," 211 EU!J;IiHh (1(1.,
Gurney and .Tac:ksoll, Loudon, 1\)32.
25. WRIGH'l', "Soil Analysis," Thomas Mllrhy and COlllll:IIlY, LOlHlol1,
1934.
26. YOE, "Photometric Chemic III Analysis, ColorinlPtl',V," ,Jolin Wilny &
Sons, Inc., New Ymk, 1928.
27. - - - , "Nephelomr,try," .Tohn Wiley &; Sons, Inc., New York, 1!J20.

References on Physical and Chemical Data

1. A'rAcK, "The Chemists YClLr Book," Chemienl Cntalog Company, Inn.,
New York, 1930.
2. and HAHN, "A Dictionary of Cho111ielLI Soluhilitios, IIlorglmi(I,"
OOMIGY
The Macmilbn COmpl111Y, New York, 1.921.
3. HODGMAN and COOI,BAUGH, "Handbook of ChOillist)'y lind r'h,VHil:H,"
The Chemical Rubber 00., Cleveland, 193!>'
4. "International Oritical Tnbles," MeGmw-HiII Book Company, Inc:.)
New York, 1926.
 IN'PRODUCTION xxv
5. LANDOI11'-BonNS'l'BIN, "Physicalisch-Ohmnisc:hc Tabcllcn," Julius
Springer, Berlin, 1923.
O. MUlmAY, "Stanchtrds and Tests for Reagent Chemicals," D. Van
Nostrand Company, Inc., New York, 1920.
7. OLSEN, "VIm Nostl'l1nd's Chcmical Annual," D. Van Nostrand Com-
pany, Ine., New York, 1928.
8. SI~rIllDr,r" "Solubilities of Inorganie and Org!111ie Chcmicl,ls," D. Van
Nostl'lLud COlUPiIIlY, New York, 1928.
9. "Smithsonian Physieal Tables," The Smithsonian Institution, W:lshillg-
1.011, D. C., HJlS.
 PART I
GENEItAI. METHODS USED IN OFFICIAL
PHARMACEUTICAL ANALYSES

The theoretical considerations and the procedures involved in

gravimetric, volumetric, and gasometric methods of analysis
are the same in quantitative pharmaceutical analysis as in quan-
titative chemical analysis. In quantitative pharmaceutical
[Lnalysis, however, the general theories of analytical chemistry
are applied to the procedures used in the analysis of the drugs,
chemieals and medicinal preparations employed in pharmacy;
chiefly to those materials official in the United States Pharma-
copoeia(U.S.P.) and National Formulary (N.F.).
 CHAPTER I

REMARKS AND GENERAL DIRECTIONS

Success as an Analyst.--To be sU!Jcessful as an analyst, the
student 111.USt realize that analytical chemistry is not simple
routine procedure. Manipulative skill acquired by experience
with the ability to follow directions under the supervision of a
skilled analyst may enable one to carry out successfully certain
mwlytieal procedures. The object of teaching quantitlttive
pharmaeeuticaJ chomistry, however, is to impart a thorough
fuudmnontal knowledge of the theory as well as the practice of
ltllalyticalmethods.
'rhe successful analyst must know what reactions are taking
pIneo during an anaJysis nnu be able to understand and apply
the theory upon which the method is dependent. The analyst
must lwquire skill of technique, pa,tience, neatness, and accuracy.
The fact should be borne in mind that analysis is not carried out
for the Hake of analysis but as a means to control the purity and
strength of substances used as medicaments. Above all, to be
Rlwcessfnl, the analyst must not only use common sense in the
laboratory but think throughout each step of the procedure.
Accuracy and Honesty.-It is absolutely essentia.l tha.t all
quantitative work be performed without even a slight loss of
material or gain of extraneous matter. Every precaution should
be t.aken to prevent. such loss or gain. All solutions and precipi-
tates should be covered to protect them from dust, whenever
possible, lmel every particle of sample, solution or precipitate
mnst be regarded as indispensable to the ultimate success of the
analysis.
Absolute integrity is demanded of every quantitative analyst.
The operator is the only person who is familiar with the entire
history of an analysis and who knows whether it is worthy of full
confidence. When confidence is lost for any definitely estab-
lished reason in the course of an analysis, operations should be
a
4 QUAN'l'I'1'A'l'IVE PIIAllMAC1W'l'ICAL CIIJiJMIS'l'/tY

discontinued at once and a ncw start made. All dotcl'min:.d,iom.;

should be made in duplicate, and the results should agre(~ closdy.
Failure to obtltin results which eheck is eonehmive oviduncl1 tlmt
at least one result is wrong and that neither can be d(~llellded U pOll.
Notebooks.-All notebooks should be of tho type dosignaterl
by the instructor. A small bound notebook whidl will slide
under the balance while weighing has proved mOl'lL satiNf:wtol'Y
in the writer's experience.
The following information relative to oneIl determination
should be included:
Date.
Object 01' title.
Experimental chta.
Reactions.
Calculations.
Results.
Remarks.

This information should be arranged in n nottt, systenmtie

manner so thn,t nnyone familiar with quantitative ttIlalysis may
understand it readily.
The date is very important in all scientific work. NUIlHll'OIlR
legal decisions have been lost through the failure' of the analyst
to affix the date at the time of analysis. The objeet or titlo
should be briefly stated. The experimental data should inel\1<1o
a complete record of all weighings and measurements set down
at the time they are made. The practice of reeol'ding datn OJl
loose pieces of paper 01' in a scmp notebook and copying t.ho
original entries into a second notebook should not be i;olul'tLt.()cl.
These data may be conveniently placed lLnd followed by tho enl-
culations on. thc left-hand page of the notebook, leaving the
right-hand page for .a concise, orderly write-up of tho entire
experiment. All reactions should be given uRing stl'uetul'al
formulae where possible. The use of logarithms for all (mlollln-
tions is highly recommended for accuracy and eeollomy of timo.
Results should always be reported as percentages nnlc:ss other-
wise directed. Remarks should include any unexpoeted develop-
ment during the analysis, explanation of error, and (Jommcnt,::;
which seem pertinent to the technique or theory involved ill
the analytical method.
 JWMARK.S AND GJ!lNEllAL VIUEC"l'IONS

Economy of Time.-Economical utilization of laboratory

hours is best twhi8ved through a preliminary study of the work
to be (lonn, followecl by H, plan for its prompt execution. There
is no Ellwh thing itl-i I< speed" in quantitative work. Rapidity
of accomplislllnent is achieved not by h[tste but by planning the
work so that two or more operations may be carried out at one
time aud especinJly by being so lLCcuratc that it will be unncces-
S[U'y to repQat it determination. The following suggestions will
save time and increase accuracy: (1)
Keep the desk scrupulously dean at
ull Limos. (2) Clean all apparatus at
onee when through using it for the
day :Lnd put it in the desk. (3)
Label all solutions, filtrates, and
precipitates systemlltically through-
out the fLlwJyLicn.l procedure. (4)
:Keep two or more operations going
at one time; thus, while cooling 11
precipitate in the desiceator, make a
wcighing, 01' while washing one
precipitate, ignite another one. (5)
Utilize all time between operations
in making calculations and writing
up experiments.
Cleaning Mixture.-A good elean- FIll. l.--Wash Lottie for diB-
Wlcu. WI1tor'.
jug mixtme may be prepltrcd by
ndtling ttpproximately 40 Gm. of commercial sodium diehro-
nil1to to 400 Gm. of comrnercial, eoncentrated sulfuric acid and
stirring the mixture. Commercittl potassium dichromate Il1n,y be
used, but the sodium dichromate is eheaper and more soluble in
coneentrated sulfuric acid.
This mixture should be used to wash all glass and porcelain
waro. The mixture is best preserved in !L thick-walled, glass-
stoppered, 500 cc. bottle.
Wash Bottles.-Wash bottles for use in quantitative work
uSlutlly consist of rubber-stoppered, flat-bottomed flasks of 500
to 1,000 ceo capacity, fitted with smoothly bent tubes and a jet
made flexible by means of a rubher joint. The jet should be
drawn out so that it will deliver a thin, even stream of liquid.
6 QUANTI'1'A'1'IVE PJIi111MA(!}W'1']CJ1L CHBMIS'1'BY

Prepare two wash bottles of the type illustrated in Fig. 1, one of

1,000 ee. aud Olle of 500 ce. ofLpa(:ity, for cold nnd hot water.
The necks of wash hottles for lise with hot water should be
wrapped with ashestoc; prtpol' 01' twine or other Imitable
material.
To borG :1 hole through [1 rubber stoppel', dip t.he end of the
borer in a strollf~ solutioll of sudium hydroxide before starting to
bore. Buil all ruL,)Jer parts in dilute sodium hydroxide for from
2 tu :~ min. [lnd wnsh them thoroughly with distilled water. Fire
polish tho ends of all glass tubing by holding th()m in the bhw
flame of :.L burner until the sharp edges lmvn beeol1lEll'Olmdocl.
"Policeman. "- The so-called "policenum" is a piece of soft;
rubber tubing fitted to the end of a glt1ss rod. rrhe rod should he
a,bout 20 cm. long with a diameter between 5 and 7 mm., and it
should have plain ltlll1ealed ends. A pieue of rubber tubing
about 3 cm. long should be fitted tightly over and projeet about
2 em. beyond the rod. The policeman is used to loosen from
the walls of the containing ves:sel adhming partielps whic:h nrc
not l'eulOvable by a stream of water from the wash bottle. A
pDlicernan should nevol' be m1ed afJ a stirring rod or a,llowcd to
rCll1ll,in in aualytienJ solutions.
Reagents.~-The reagents used in quantitative analysis must
be pure. Hen,gents which conform to the requirements of the
United States Pharmacopoeia and of the National Ji'Ol'lllUbry
should be used in all assays whiGh have for their purpose iillO
establishment of the strength or quality of official substLtneeil.
Those reagents whieh meet the specifieatiolls of the Amol'ie:tn
Chemical Society as well ns those sold by repntable manufuet.mel'H
as "annlysed" or ,( reagent" gmde will usually be found to moot
all ofIieiall'eqniremonts.
Purity and Strength Requirements.-The purity and strength
of chemieals and drugs of the U.S.P. and of the N.P. are usually
expressed in terms of pc)' cent. The offieial requirements of
purity and strength obtain only when the offieh~l methods of
assay arc employed. 'When the pel' (lent strength or purity is
given in round llumbers, it is understood that the figures repre~
sent exactly the minimum 01' maximum requirement, e.g., the
requirement that hydrochloric acid contain "not less than 35 pel'
cent and not more than 37 pel' cent of Hel" permits a product of
 REMARKS AND GENERAL DIRECTIONS 7
35 or 37 per cent or any intermediate strength; in other words, the
figures arc inclusive. It is established by rule that the figures
35 and 37, respectively, have the same significance as the figures
35.00 and 37.00.
Materials Required.-rrhe statement of materials required
in the exercises is intended as a g~lidc for the instruel;or in ordering
supplies. Commonly used Teagents and solutions such as indi-
cators, acids, and alkalies are not rtlways listed since it is assumed
that they are I1v[Lilnble in the analyticalla,bomtory. The quan-
tities indicated arc fo1' each sample. It is possible to give only
I1PPl'Oximate quantities for materials used as solvents, [or washing,
to muke alkaline, and so forth. All reagent materials should
meet the official requirements of purity [LIlel strength.

SAMPLING

The first important consideration in an analysis is the securing

of a representative sample, a consiclemtion too freqnently
ignored by analysts. Fixed rules cannot be laid down in sam-
pling for much depends on the IUtture of the materiaJ and the
quantity from which it l'epresentntive Rumple is to be taken.
Whenever possible, a sample fI:om each package should be exam-
ined separately. "\lI[hen a single package is sampled, the contents
should be mixed or samples should be tfLken from different parts
of the container and mixed, and then a portion of this mixture
should be taken for analysis. The sampling of bulk quantities
of chemicals and drugs lllay be perf onncd by the methods given
ill the U.S.P. for sampling vegetable tlrugs:
"1. It is recommended that gross samples of vegetn,ble drugs
in which the component parts are 1 cm. or less in allY dimension,
and all powdered or ground drugs, be taken by means of a sampler
which removes a core from the top to the bottom of the container,
not less than two cores being taken in opposite directions; that,
when the total woight of the drug to be sampled is less than
100 kilos (200 pounds) at least 250 Gm. shall constitute an oJJlcial
sample; that when the total weight of the drug to be sampled is in
excess of 100 kilos, repeated samples shall he taken by the above
method, and accor~iing to the schedule given below, mixed and
quartereeI, two of the diagonal quarters being rejected, the
.3 ()UAN'l'I1'A'l'IVE I'IIARMAC'EU'l'[(!AL CIlEMISTRY

remaining two qut),rters being combined and cnreflllly mixcd, and

u,gain subjected to a quartering prooess in the same manncr until
two of the quarters weigh at least 250 Gm., whieh latter quarters
Elhall constitute an official sample.
"II. It is rccommended t,lH1t p;ross samples of vep;ctnhle drugs
in which the c:omponent parts [ire over 1 em. in any dimension
be taken by hand; that when the totnl weight of the (lrug to ho
sampled is loss than 100 kilos, at least [iOC) Om. shall cOIl:,;tituto
an oJJicial sample, and this Rllall he taknn fronl differnll t, part:,; of
the containor or eontainerR; that whon the j;otftl wnight of the
drug to be sampled is in ""XeORS of 100 kilos, rnpentcd :,;:tmpIOR
shall be taken by Llw above method and aeeording to the s(:lindllln
below, mixed aud qnartered, two of the dit1gonal quart(mo; lwing
rejected, and the remaining two qunrters being; eombiUfHl and
earefully mixed, and again subjeeted to a quartming process
in the same mRll1ler until two of the quarters weigh not loss than
500 Om., which latter quarters sha.1l constitute an o.[ficial sample.

SClmJ)lTLm RJ"COMMElNllBIl Fon SAI\[PJ,ING

Numbor of P!1[:lmges Number of Paelmges
in Shipment to ne S:unplml
1 to 10 1 to 3
10 to 25 2 to 4
25 to 50 a to G
50 to 75 to 8
fj
75 to 100 8 to 10

"When over 100, the total number sampled should not he

less than 10.
"III. When the total weight of a drug to be sampled is less
than 10 kilos, it is reeommended that the above Inethods he
followed but that somewhat smaller qnantities he withdmwll, and
in no case should the final o:f!icia.l sample weigh less thfm 125 Om..
,e IV. In addition to the withdrawing of o.Uicial sarnplc8 :weorcl-
iug to methods I, iI, and III, the o:{/icial s(Lrnplo mn,y eOllsist of
the total amount of a direct purehase made by Fedeml, State or
Municipal Food and Drugs' Act enforcement officials."
Students' samples are supplied ready for !Lll!11ysis as a rule.
Samples which require drying should be spread on a large watch
glass, thoroughly mixed, and dried in an oven at the specified
 RBMARKS AND GENERAL DIIlEC'l'IONS 9
temperature. They should then be transferred to a weighing
bottle and kept in the desiccator.

CALCULATION OF RESULTS AND ERRORS

Source and Nature of Errol's.-It is only in rare cases that the

numerical value of an cxperimental result can be directly deter-
mined. As a rule, it is necessary to calculate the result from the
different measurements and observntions whieh have been rru1de.
Duplicate results which check very closely are not an assurance
of aecuracy, as is often taken for granted by the beginner in
qunntit[1tive work, since errors inherent in the method and
apparatus used may be roveated for each sample. Generally,
results which agree closely when obtained by two different meth-
ods of an:11ysis are It good indication of the reliability of the
methods.
The results of repCl1ted analyses or measurements will fail to
a,grec, in general, when made by the same 01' different analysts
to the full precision of which the method or instrument is capable.
The discrepancies in the results tUG ct1Used by various sources of
error t.o which all experimental data are subject. The errors
are of two types, namely: indeterminate or accidental errors
which manifest t.hemselves by slight variations in a series of
observations made by the same observer under identical condi-
tions. They result from causes difficult to detect, such as differ-
ences in the judgment and skill of the analyst. Generally,
indeterminate errors are intangible and their climination by the
analyst is impossible. Determinate or constant errors are of
such nature that they recur in a constant manner in each of a
series of determinations. Consequently, it is possible partially to
determine their value and reduce t.heir effect on the final result.
They arise from cnuses such n,s: (1) Personal errors made by the
individual analyst, c.g., inability t.o judge color changes sharply,
resulting in habitual l'elLding of end points in titration too late.
(2) Errors of method caused by faulty procedure, e.g., incorrect
sampling, contamination of precipitates, and improper selection
of indicators. (3) Apparatus errors due to poor construction or
calibration, e.g., inaccuracy in the calibration of burettes or
pipettes, inequality in the length of the arms of the balance, and
10 QUANn'1'A'1']V}I} 1'1lARMACliJU,]']CAL Cl1liJMIS1'UY

incorrect weights. Errors of this type nrc wmally detne(;ablo

and so m~w be eliminttLml to tL brgn extent.
Indeterminate Errors. -~Indetel'milHLL() ()l'l'ors follow tho law
of clU:LllCe which is represented by the followillg (ntl'V!) (:omlllonly
known as the cllrve of CI,),O), or tlto probability (;UI'V!:.
An examination of this curve Ahows that: (1) Very large errOl'H
are unlikely to occur often; (2) HlliaU Ort'01'H O(l(!lU' witJl l,!;l'uaLel'
frcquelley than bXl!;e ones; nnd (:1) positive :mu negative elTon;
of the same nUll1ericalllln,gnitude me equnlly probuhle Ainee the
curve is sYHlInetricn,l with respect to the Y axis. COlulequontly,
y

I'IlI. 2.-Pl'obllbilit~y cmrve.

jt is apparent that the best value to select for n series of observa-

tions is their 11rithmetiealmean.
Average Deviation from the Arithmetical Mean.---In order to
measure the magnitude of indetcrminn,te or aecidentn,l OWn'S,
use is made of the theorem proved by the method of least squares
which statos that an arithmetical mean computed from n equally
trustworthy ohservations is -vn
times as trustworthy as nny
single observation. If the indeterminate error in allY one obser-
vation is represented by i, the indeterminate error of the arith-
metical mean is iI-VIi; in other words, the uncertainty of tho
arithmetical mean is inversely proportional to tho Hqllare root
of the number of observations. The devirLtion of a singlo meUH-
momont from tho arithmetical menn of a series of similar mOI1H-
mements cnn readily be determined and from this the [Lvoragn
deviation of a single determination may be ea,leulnted. For
example: (1) In the standardization of hyclroehlorie add by
titration against pure sodium carbonate, the burette readingH
30.25, 30.22, 30.26, 30.27, and 30.23 ee. were obtained. The
 REMARKS AND GENERAL DIREC'lTIONS 11

arithmetical moan of these numbers is 30.246. The deviations

of the individual readings from the mottn, neglecting sign, me
0.004, 0.026, 0.014, 0.024, anrl 0.016. The sum of these five
numbers is 0.084, and the :wcrlLge is 0.084/5 = 0.017. Express-
ing the average deviation as [L ratio to the arithmetical mean
gives 0.017 part per 30.240 or about 0.56 part per 1,000 parts.
(2) Samples of tL pure ehloricle weighing 0.5056 am. yielcled cal-
culated results in grams of 0.<1850, 0.4840, 0.4844, 0.4846, and
0.4887, when the chloride ion was precipitated as silver chloride.
Calculate the inaceurlwy (absolute crrul') in pel' cent nnd the
lllean error in parts per thousand from the rosults. The mean
value obtained by experirnent and caleulntion, rejccting the last
detcrmination (sec Rejection of n Result), equals OA845. Tlw
absolute error is 0.5056 - 0.4845 = 0.0211. The relative error
of the method is 0.0211/0.5056 X 100 = 4 per oent (t\pprox-
imate). The mean errol' of the result is 0.06 pel' cent or 0.6 part
per 1,000 parts.
The example shows that the inaccuracy of the method used is
40 parts per 1,000 parts as compared with a mean errol' in the
determinations of 0.6 part per 1,000 parts.
Rejection of a Result.-Sometimcs in a series of detennina~
tions, one of the results obtained will differ greatly from the
others without any apparent mistake having been ~nade in the
work. The questioll then [Lrises as to whether this result should
be retained with the others in caleulating the value of the arith-
metical mean. If at least four determinations have been made,
the result in which tl18 error appears to be large rnuy be omitted
and the arithmetical mean and the average deviation of the
other results computed. If the difference between the arith~
metical ll1.can and the result believed to be in error is foUl' or
more times the avcrlJ,ge deviation, the result should be rejected,
e.g., in the preceding example, the last resl1lt should be rejected
since 0.4887 - 0.4845 = 0.0042 and this difference is more th:m
four times the average deviation, 0.0003. It hflS been shown that
the deviation of a result which is four or more times the average
deviation is clue to a 111.istake in all except about five cases in a
thousand.
Significant Figures.-Significant figures arc the digits which
when placed in order give the value to a number, e.g., in the
12 QUANTITATIVE PHARMACEUTICAL CIIEMI81'UY

quantity represented by the number 236, the digits 2, 3, !tnd 6

are significant figures. Zeros are employed in some cases to
locate the decimal point; in other cases, they mu,y be significant
figures. Thus in the number 20.5, the zero is a significant iignrc,
but in the number 0.0125 the zeros are not significant but silow
the order of magnitude of the other digits. In the qn:1nti['y,
1.0000 Gm., the zeros nrc significant since they indica[,e that the
weighing h:1s been made to YIo mg. and that only the last zero
is uncertain.
Every measurement involves some error or cun be carried out
only with a certain degree of [1ecuracy. Consequently, it is
evident that the number expressing the value of a Inet1suronHm1,
can be only approximate. Irrespective of the number of figmos
written as the result of a calculation, thc accuracy of n, valno
beyond that limited by the errors oC the me(1S\lrelllcnts et1lmot
be increased. On the other hand, if too few figures are written
down, the resulting value may be ll1uoh less 110Cllrate thn,n the
data permit. The proper manner of expressing Lt result is to
retain or use such a number of figures that all, exeept tho bst,
are khown with certainty, and that while the last figure is Ulleel"-
tain the error is not greater than 5 in the following plaeo.
For example: If a burette can be reftd to 0.01 co. and 32.5 instead
of 32.50 is written as the result of a measurement, the mistake of
writing too few figures is committed since the number H2.5
indicates that the true value lies between 32.45 and :32.55 and
the apparent error is five times greator than the error of measure-
ment. Again if, in recording the result of n, series of bmnUe
readings, the mean of the values 32.52, 32.54, 32.54, and ;{2.55 eo.
is written as 32.538, it would be a mistake of retaining too many
figures since it would indicate that the error of moasurmnolli; is
only about 0.0005 cc. The number should he rounded off,
therefore, to 32.54 cc. For purposes of further caleulntioll, all
of the figures should be retained, i.e., the number i32.538 sllOulcl
be used.
In general, for l1na.lytical work, the op(!mtioILs of addition and
subtraction should give as the finul result no more (keimal
places than the least llumber of deeimal places eat.milll,!; iut.o
the calculatioll. This is illustrated by the following ()x:ullplo:
In the addition 12.4 + +
121.502 3.6653 = 137.5673, the mnxi-
 llEMAlU(S AND GENEllAL DIR.lfJC'l'JONS 13

mum apparent errol' in the result is 0.05 whieh is the great-

est apparent error in the number 12.4. Therefore, a derived
01'1'01' of 5 exists in the seeond decimal place, figure 6, of the
result; :)'lld the result should be written 137.6 but if the result is
used for further c[Llcubtions, it is better to retain one figure more
'i.e., to omploy 137.57.
In multiplication or division, as a rule, retain one m.ore sig-
nificant figure, when available, in the result than there are in the
nnmber having the least number of signifieant figures. For
example: The product of 11.32 X 12.2 X 0.0321 = 4.4331384.
If the bst figure in each number is uncertain, the relative errOT
in the number 12.2 will be about 5 in 1,200 or 0.4 per cent.
Consequently, it would be incorrect to write the result as
4:.433138!t for this result has a derived error of 0.4 per cent or
of about 2 units in the second place of the decimals. All figures
following this are therefore meaningless and should be discarded,
the result being written 4.43.
Logarithms and Their Use.-The solution of problems and
the calcubtioll of the results of analyses from data are greatly
faciliLated by the use of logluithms. It is advisable for studente
to make calculations by the use of logarithms in order to save
time and avoid teclious calculations by long hand.
A logarithm is an exponent which must be applied to a fixed
positive number other than 0 or 1 to produce Hny given number.
The fixed number is called the base. In the common 01' Briggsian
system of logarithms, the base is 10. A logarithm consists of
two parts, the characteristic or integral part and the mantissa or
fractional part. The characteristic of n. number greater than 1 is
always one less than the number of digit.s to the left of the decimal
point, 1:.e., where n equals the number of digits to the left of the
decimal point, t.he charact.eristic is represented by a number
equal to n - 1. For example, the characteristic of 10,000 or
10 4 is 4, of 100 or 102 is 2, and of 1 or 10 is O. The characteristic
may be either positive or negative. When there are no digits
t.o the left of the decimal point, the characteristic is negative or
minus, and when there are no zeros before the significant figures
to the right of t.he decimal point, the characteristic is -1 or
9 - 10. When there are no digits to the left of the decimal
point and n zeros to the right of the decimal point before the
14 QUANTI'l'A'l'IVE PlIAIUvIACEUTICAL ClIRM18'l'NY

significant figures, the charactcristic is en -

11.) - 10, c.g., the
characteristic of 0.1 is I or 9 - 10, of 0.01 is 2' or 8 - 10, tLlul
of O.OO()OOl is 6 or 4 - 10.
The mantissas of numbers arc fmnished by th(~ table of log~
al'ithms, see pn,ge 45:3. The ext1Inplcs given below illnstmte
tho use of the tltbles:
E'J:ample L--Fincl the logarithm of 94G5. Frum Ow explana-
tion given, it is eviclell.t tllltt the chanwtnristie is 3. '1'0 fmd tho
mantissa, locato 94 in the logn.rithrn tahle in tho eolumll of
Natural Numbers iLnd then move dil'ecLly to the right, to the
colurnn uncleI' 6 at the top of the page. Tho malltiRsa given there
is 9759. This is tho mantissa for 94GO. To find the Illnntis,m
for 9465, move [tu'thcr to the right to the eolumns headed
Proportional Pnl'ts where, undor the 5 eolumll, tho iigul'u 2 is
given. This figure 0002 when n,ddecl to 9759 shows how n1.1.wh
more the logarithm of 946.5 is than Uwt of 9400. Consequently,
the logarithm of 9465 is 3.9759 +
0.0002 = :3.9701..
E:cample 2.-Find tho logarithm of 0.0001058. Sinee tho 1i1'...,t
significttut digit is four pitwes to th() right of the deeinlnl point, the
characteristic is 6 - 10. ]'ind the rnrmtissa aN in ex:tlHIJle 1 by
following the horizontal line oppusite 10 to the vertiea'! eoluwn
headed by 5 and add to this the proportional part UtHIul' the
column headed 8. The mantissa is then found to be 0212 .+.
0033 = 0245 and the logarithm is wdttnn 6.0245 - 10.
EXfLmple 3.---Multiplyillg and dividing with logarithms.
To llluitiply, iLdd the logarithms of j;he numbers together. To
divide subtract one logarithm from the other. Study tho follow-
ing example: .

24.4() X 0.2970
1000 X 0.003741

Multiply by adding the logarithms

log 24.46. . . . . . . . . . . . . . . . . . . . .. = 1. :3881)

log 0.2970. . . . . . . . . . . . . . . . . . . .. = 9.4728 - 10

24.46 X 0.2970 is the sum of the

logs ........................ = 10.8613 - 10
 REMARKS AND GENERAL DIRECTIONS 15
log 1000 ..................... . 3.0000
log 0.003741 .................. . 7.5730-10

1000 X 0.003741 is the sum of

the logs ............. '" ..... = 10.5730 - 10
Divide by subtraeting the logs

10.8613 - 10
10.5730 - 10

0.2883

The result of these operations is the logarithm of the number

sought. To find the nlLtuwl number eo.rl'esponding to the
logarithm 0.2883, turn to the table of antilogarithms (pllge 455),
move down the column headed Logarithms until 28 is found,
~nove horizontally to the column headed 8 where the number 194i
is found, and the11 procced to the Pl'Oportional PartB column
under 3 where the Humber 0001 is found; add this to 1941, the
sum is H)42. The chamctel'istie is O. Therefore, 011e digit is
to the left of the decimal point and the reqllireclllUmbel' is 1.942.
Questions and Problems
1. Rewrite eaeh of the following q\lantitic~, underscoring the significll,llt
1\g\ll'O~: (It) 1.1200 Gm. weighed on the analytical balrLllcc, (b) 20,459 ce.
from a burette reading, (c) 0.0062 Grn. frolll a weighing on the analytical
balance.
2. Make the following e:l}euhtions ret.aining only the significant figures:
a. 2.3 + 100.50 + 4.27:3 - 20.005
b. lOO X O.OlD X 1000 7- 10 =
c. (20 X 1.0250) - (10 X 1.2000) 7- 1.200 =
3. A sample known to be \)8.,50 per cent pure yielded results on analysis
in duplic:ate of 98.30 and !JS.12 pCI' cont, l'e~pectivoly. What is t,he precision
in pnrts pCI' thousand of each determination?
4. The result of an analysis for per cent purity employing the usual
appamtus nnd weighing to the fourth dccimfll place was reported as
86.3584323 pel' cent. How should the resuH be reported?
6. The followillJlj results were obtained in the determination 01 chloride
ion in a s[\mple of pure potassium chloritle:.47,48, 47.56, 47.50, 47.62 and
48.25 pel' cent. Calcuhtc: (a) the mean deviation, (b) the pel' cent deviatioll,
(c) the meHn deviHtion in parts pOl' thousand, (d) tho mean error in per C01lt.
Should Hny of the resn] ts be rejected? Why?
16 QUANTI1'ATIVE PHARMACEUTICAL CIiEMIS'1'RY

6. Find the common logarithms of the following numbers: 100, (j5, 8M2,
0.221, 0.00018.
7. Find the numbers (antilogarithms) which corre:>pouu to oneil of t Iw
following logarithms: 3.5448, 0.8250, 0.1260, 2.3fl7\).
8. Usillg logarithms, multiply: (a) 21500 X 0.000:332, (II) 0.00,18)<
0.0008'126, (c) (42.113 - 1.85)(30.20 + 12.82)(0.5444 X 0.1112).
9. Using logarithms, divide: Ca) 0.01648 by 0.\)'172, Cb) 20.0,1 by 110.80,
(c) 1020 by 12.64.

GENERAL OPERATIONS

Crucibles.-A variety of crucibles arc used in (l\lanLiLlttive

analysis. Those most comlllonly employed are lluLcle of high~
grade porcelain. They withsLttnd high
tempemtures and are suitable for use
in the ignition of ,most; drngs nnd
precipitates. They me no!; imitable
ror fusions beCltUse the gbze ifl aLLankml
by the flux, especially if the flux is
basic. Other cl'llcilllc:-:l eousist of fmiCd
silica, alundum, nick,el, ancl plntinllm.
The Gooch Filtration Crucible. !1'1w
Gooch filtmtion cnwiblc (Fig. ;j) is
designecl for t.ho sepanLt;ion of p]'peipi~
tates by suction filtration. It !tas :L
perforated bot.tom upon which is
bedded a mat of asbestos, tlms lllak~
li'lG. a.-Diagram of Gooch ing it possible to collect, WHflh, dry,
crueihle assembly lot" fUtra-
tion. and w(~igh a pl'eoipitate in tho same
crucible.
Preparation and Use of Gooch Crucible.--Obtttin rLllOnt 0.0
Gm. of [tRbestos fiher (washed with hyciroehlol'l(l ncicl) nlld fllUiko
it thoroughly with 100 cc. of distilled water in n sllitahle Iln..,k.
Allow the asbestos to settle and decant most of tho water ()()n~
taining the fine fibers. Add about. }OO co. of wld;er and agitate
the mixture again. Fit the Gooeh funnel contailling the erueible
in tbe mouth of a filter flask connected to n sllction pmllll. 1'0111'
the suspended asbestos mixture into the cl'ueiblc ,in small SlWees-
sive portions applying suction gently tlfter tho n,ddition oj' e[tcb
portion until a smooth mat not over 1.5 nUll. thick is obtuined.
Upon holding the crucible to a strong light, the holes in the
 nIiJMAllKS AND GENERAL DIRECTIONS 17

bottom should be just pereeptible and the mat of asbestos should

appear uniform. Cover the asbestos with [L perforated porcelain
diRk (filter plate) and add a very thin layer of asbestos. Wash
the filter mat with 200 to 300 cc. of water or until no trace of
aflbestos fiber passes into the washing. Place the prepared
crucible on a waLeh glass and dry it for 1 hr. in an oven fit 120 to
130. Allow the crucible to cool in a desiccator, then weigh.
Hepcat until the weight of the prepared crueible is constant
within 0.0002 Gm. for two suceessive weighillgS.
To use the Gooch crucible, place it ill the funnel and apply
moderate suction. Pour the liquid from the precipitate into
the crllcible, wash the precipitate in the beaker by decantation,
transfer the precipitate to thc crucible and
complete the washing. The rocl used to
guido the liquid into the crucible should
ncver be allowed to come in contact with
the asbestos mat, since it may break the
thin film above the filter plate. The filtrate
should be examined for traces of asbestos Fw. 4.-FritLed-glass
eruciblcs, low form und
fiber, and if sueh lue fo-und, it should be high form.
repassed through the filter.
A properly prepared Gooch filter may be used for a number of
sueeessive dotcrminations where the precipitates lue coarse
grained and crystalline.
Fritted-glass Crucibles.-Fritted-glass crucibles (Fig. 4) have
fused in sinterecl or fritted-glass bottoms and are supplied in two
porosities, "medium" and" fine." Crucibles of medium porosity
are suitable for the filtration of flocculent and moderately fine
pl'eeipitates. Crucibles of fine porosity are suitable for the
filtration of fine precipitates such as mercuric 01' bisl11.uth sulfides.
These crucibles are useful in analyses where the residue is to be
dried to constant weight at temperatures below 150C. 'rhe
crucibles should be heated in an oven and allowed to cool to
room temperature in a desiccator.
Filtration and Washing Precipitates.-The most usual and
efficient method of washing a precipitate is by the process of
decantation. The liquid above the precipitate is poured along
a guide rod onto a prepared filter, some of the liquid and most
of the precipitate being allowed to remain in the container.
18 (JUANTn'.A'l'lVE' PlIARMAClW'l'ICAL OJ1BMIS'l'RY

A given amount of wash water is tllOn poured on tho precipiLnte

and stilTed vigoroLlsly. 'rhe precipitate is allowed to set He,
a,nd the sup0l'llat,f\,nL liquid is dOC[lllLed u,s bdore. This process
is repeated three or four times. The rll"Cwipitatc is tlwn washed
onto the filter, Hsing a "polieemfLll" if neeCSf:il11'Y to dislodge
any partidos whidl adhere to the container wall. Tho filter
containing the pl'odpitato is was hod by emlRing a fine HtrOlt1ll of
liquid f['oin tho wash bottle to fall upon the filter, nlwnYH washing
from the top edge down, until the filter is about three-fourths
full. The filter is allowed to drain until pnwtieally ompty, Lhon
the rinsillg is repented. After about five slwh rinsings, 1 or
2 cc. of the final filtrate should be tested for complctencHs of
wnshing.
It should be borne in mind that it is better to wash 11 pweipiLate
a eomparatively large num.ber of times with small portions of
liquid iJHLn only a few times with brge portiOllfl and that. eueh
portioll of wash liquicl should be removed as eOllllllotcly I1S
practicable beforo tho next portion is added. Nevor fill the Jiltt~r
more than to within one-fourth inch of its upper edge. The size
of the filter should be governed by the amount of precipitate,
not by the amount of liquid to be filtered.
Colloidal and Fine-grained Precipitates.--Oolloidal precipi-
tates, some of them gelatinous, some of them finely divided,
Dxe given by the most insoluble substances, such as sulfideR,
hydroxides, elemcmt11ry sulfur, etc. Although insoluble, they
have the property of being converted by water into colloidal
suspensions which pass through the filter. These ImbstancOfl
can usually be coagulated and rendered filterable by boiling
them after tho addition of' an eleetrolyte, such as NH.1NO a,
NH-lCl, or Hel. It is in pmt for this reaSOll tha:l', prepared wash
liquors, rather than pure water, are used for washing certain of
these precipit:1tes.
AU crystallization is preceded by supersaturation. If super-
satumtion is high, crystallization will be spontaneous and tlte
crysta,ls formed will be so fine that they will pass through the
filter. If the preeipitating agent is acldecl slowly with constant
stirring, high local supersaturation will be avoided, and the
crystals formed will grow in size to such an extent that they will
be retained by the filter. By using different concentrations, it
 BEMARJ(S AND GENERAL DIRECTIONS 19

is possible to get the same salt crystallized out in crystals varying

in size from full-faced crystals down to the colloidal form.
Evaporation of Liquids.-The evaporation of liquids is best
carried out in porcelain dishes constructed for that purpose which
expose l~ large surface of liquid to the air. Evaporations should,
with few exceptions, be rlttl'l'icd out, on the water bath to avoid
dangor of loss of materinl due to spattering or bumping and to
avoid deeOlTlllOsition. The dish should be covered by an inverted
funnd supportpd above it or with a watch glass elevated above
the rim of the dish by means of a glass triangle to prevent the
entry of foreign lIla,ttel' into the liqllid. Large quantities of
liquid are best evnpol'ated in a comparatively small dish by
ndding fresh portions of the original liquid from time to time
to replace that lost by evaporation. This helps to eIim-
hUtte the source of error involved in the frequent transfer of
liquids.
Transfer of Liquids.-As few transfers of liquid as possible
should be made, since eaell permits of a possible source of error.
When transfers are necessary, they must be made quantitatively,
washing the original container with successive small portions of
wash liquid. This procedure is the most efficient, since it most
thoroughly removes the rnaterinl and keeps the volume of liquid
within workable limits.
When tru.nsferring liquids from one vessel to another, a guide
rod should always be used. It should be held tightly against
the lip of the (Jontainer.
Drying and Ignition of Samples and Precipitates.-Many
substances are direc1led to be dried under standard conditions
before analysis to correct for absorbed moisture. Precipitates
frequently must be dried previous to ignition. N umel'OUS
types of drying ovens are marketed which are suitable for this
purpose, the electrim111y hen.teLl ones being best suited to constant
temperature control. Sl1mples should be dried at the specified
temperature on a watch glass 01' in a suitable container. Precipi~
tates may be dried directly on a filter by placing the funnel
containing both direetly in a drying oven adjusted to a telll~
perature of 90 to 100 unless otherwise specified. The funnel
should be covered with a sheet of filter paper held in place by
crimping its edges over the rim of the funnel.
20 QUAN'l'I'l'A7.'ll'E PIIAW,{JlC1W'l'ICAL CllElI[[Sl'IlY

Preeipitlttes may frequently be ignited without previous

drying. To do this, the precipitate is folded within the filter
paper, and the whole pbcecl in n vertiettl crucible. Tho m'ueiblo
is covered and heateel gontly by iutennit.tcmt applicatiou of tho
flame until all of the moiRtuJ'e is driv(m. off anel the pa[lm' iM
well clmrrcd. The nover is then rmlloved, and tho t:l"Iwihle
placed in an inclined position on the triang;le (Fig;. fi). Tlw
full flame is then turned on and ignition continued until all CU,l'bOll

FIG. 5.-Posit.ion of crucible above Harne during ignition.

has been burned away, rotating the crueibJo sligh L1y from Limo
to time to expose fresh portions uf precipitate. The c:nwihlo
containing the ignited rer:;idue is allowed to enol in n dOfiieeator
and weighed. It is then ignited n. second time and weigholl. If
the successive weighings do not f),gree within 0.2 mg. ,,,hon n
porcelain eruciblc is used, it should be ignite(l :md wnigh(J(l l1
third time. These opel'lltions shollld. hn repeatc'cl unt.il (:Ollst:mt
weight is attnineU.
Constant Weight.-Tho term dr'icil to constant wldaht monllS
that two consecutive weighings do not differ by more Lllan 0.1
per cent when the second weighing is made aftel' an mlcli.tional
hour of dryin/!.
 [WJMARJ(S AND GENERAL DIREC'l'IONS 21

Use of Desiccators.-Desiccators are a special form of

glass vessel, rendered air-tight by means of ground contact sur-
faces, used to maintain l1 dry atmosphere for objects that i:night
be affeeted by moisture or carbon dioxide. If the contact sur-
faces are not air-tight, they may be made so by coating them
with a thin layer of grease prepa,red by melting together equal
parts of beeswax and petrointum. The desiccator is usually
divided into two compartments separated by a perforated por-
celain plate. The lower compartment is charged with a delty-
dmting agent, 011e of the most comlllon of whieh is anhydrous
cn.leium chloride, but concentrated sulfuric acid, solid potassiulll
hydroxide, unslaked lime, magnesium perchlorate, etc., Ine
HOlnet.imes used. The porcelain plute is usually fitted with
holos for erucibles. The desicca(;or should always be kept closed
when not tmnsferring crucibles into or from it ..
THE ANALYTICAL BALANCE
A knowledge of the principles and use of the analytical balance
(Fig. ('\) is essential in all quantitat.ive chemistry. The diagram
(Fig, 7) SllOWH the variolls rmrts of sllch a bahnce with the propel'
name of each part. Other bl1111,nces on the market differ slightly
in construction but the pm-ts arc essentially the same for all.
The ttllalytical balance is a delicate instrument of precision which
must be handled with extreme care to ml1intain its fine adjust-
m(mt and aCOll1'aey. Each student under the direct supervision
of 1111 im;tnwtor should learn the name and function of each part.
The following brief comments will serve to assist the student
better to uuderstand the function and care of the balance: The
balance beam is supported on a center knife edge or fulcrum,
usually of agl1te, which rests upon an agate plate. Two pallS
intended to support the masses to be compared are suspended
vertically from each end of the beam by stirrups. Each stirrup
has an agate bearing whieh rests on a knife edge. The arms of
the ba,htneo beam arc gmduated so that a rider, of known weight,
ean be pIn,cocl at any desired di:,;tanee from the central knife
edge. Forcible or SlHlclcll lowering of' the beam npon tho knife
edges dulls them [wd produces furrows in the bearings which
decrease the sensitiveness of the balance. A pan arrest is
provided which when pushed in and turned releases the pans,
22 QUANTI7'ATIVE PHARMACEUTICAL CHEMISTRY

FIG. 6.-A typical analytioal bahIIlC(l.

Rm'Bll. 'Rot) CA'Rnl'll. ----.

CgNTRE }(NIFE EOGE ----'-" \

STJRRUP--__
BEAMARRE5T--
'STIRRllP HOOK---
GRAVITY WEICHT
_ -----COl.UMN
____ INDICATOR
[QJl.NEr.DU.]

__ ----LEVEL

____ PAN ARREST

---INOEX Pt.ATll

:FIG. 7.-Diagram showinQ; p!lrts of tho analytical bailmco.

 llEMABKS AND GENERAL DIRECTIONS 23

leaving them sUl-lpcncled from the sLirrup. A long pointer which

multiplies the roi;u,tional displacement is attached to the beam in
order to relluer small movements of the beam perceptible.
Rules for the Use and Care of the Balance:
1. Use the babrwe assigned by the instructor.
2. Adopt an attitude of personal responsibility for the condi-
tion of your balance; the carelessness of one student may render
ilUlCcumte the work of all who use the same balance.
3. Brush the pans and floor of the balance with a camel's-hair
brush before starting to weigh any substance.
4. 'Test the adjustment of the balance before each weighing.
The balance is properly ndjusted only if the following conditions
are fulfilled: (a) The balance is level as shown by the spirit
level; (b) the mechanism for releasing and arresting the beam
works freely and smoothly; (c) the pan arrests just touch the
pans when the beam is lowered; Cd) the pointer rests at zero
when the beam is either released or arrested while the pans are
supported; and (c) the pointer swings equal distances on either
side of zero on the index plate when the beam is set in motion
without fI, l(md on the pans or with an equal load on each pan.
Allowf1lwe may be made for a variation of one division on the
index seale without adjustment.
5. Do not attempt to adjust the balance. Call an instructor
if it requires adjustment.
6. No sample should ever be placed directly upon the balance
pan. Solids may be weighed on a watch glass or in a weighi.ng
bottle, and liquids must alwnys be weighed in a tightly stoppered
bottle. The nse of sheets of paper in place of a proper container
is not permissible.
7. Hot objects must be allowed to cool to room temperature
before placing them inside the balance case, since currents of
warm ail' tend to buoy up one a,rm of the balance and also to
cause thtlt arm to expand in length.
8. Mn,tcrial spilled on the balance pans or floor must be
brushed up at once.
9. The babnce door must he closed when the final weighing is
made.
10. 'The beam should be set in motion by gently lowering it
so that the pans are supported by the pan arrests; the pan arrests
24 QUAN"PNA'l'IVN PHAIUt1AC]i}U'l'JOAJ~ CllRM18'/'RY

arc then Cflrefully lowered, and if the beam does noi; swing, it
may be sei; in motioll by mea,m; of the ride!'. Long swings of tho
point-or are not necessary. The point(H' ShOllld swing through nil
amplitude of not less tluLil two divisions to ()ith(~r si(le of the >ler\)
of the index scttle.
11. All weights should be handled with the fOl'eeps; they should
never be touched with the fingers. PIneo IHltLvy woights in the
ccnter of the pan to prevcnt oseillntioll.
12. Record all weights directly into your notebook npon l:om-
pletion of a weighing. Before removing weights, ehock thl)
balance by arresting and releasing tIle bemn anel be sure that
correct balance has been al;Lniucd. ltmnmnbor th:tt a Hlight ClrrOl'
in weighing may render worthless un fUl'ther annJYHis of It smllple.
13. Before leaving the balance, stop the motion of the beam
by means of the pan arrests, raise the benm off tho agate kllifn
edges, be sure the balance is clean, and CIOBC the door of tho
bahmce case.
Sensitivity of the Analytical Balance.--The senHitivity V[1l'ies
inversely with the weight of the beam. The lighter the beam,
therefore, the greater the sf1nsitivity. The sensitivity vl1ries
with the length of the beam. The sensitivity varies directly to
the time period of oscillation. The sensitivity vnries inversely
with the load on the pans. The sensitivity of a balance may bo
varied by moving the small gravity woight on the pointer upward
or downward, thus ellftnging the position of the center of gmvity
of the beam. The efficient operation and sensitivity of It balance
are dependent upon minimum friction between the ngate
knife edges and bearingR.
Exercise 1

Object.-Determination _of the True Zero Point of a Balnnoe.

Procedure.-Dust the pans and floor of the Laiunce with It (lamol'A-hair
brush. Close the bui::lllce door. Gent.ly release the beam 11lld pltn tnreat:,!,
avoiding setting the beam in motion. Cause the beam to swing so tiw,t the
pointer moves past not more than seven or lesH tllltll two divisiolls of tho
index scale by carefully touching one arm of the beam with the rider.
Allow the pointer to complete two oscillations without taking a l'eadillg.
Then take three readings of the limit of pointer oscilhLtion on one sirln
of the index scale and two on the other, counting first left then right. Take
the average reading for each side, find the difference between the aVel'llgeH,
 REMARKS AND GENERAL DIRECTIONS 25

:tnd tlivide the difiereIH:e by two; the result gives the true zero point of the
balltllce.

The zero point is then computed as follows:

Left Right
5.2 4.6
5.1 4.4
5.0
:3)15.3 2)9.0
5. 1 4.5 average
5. 1 - 4. 5 = O. 6 difference

0.6/2 = 0.3 of [l, sonlo division to the left which is the true zero
point. Ma,kc al; least three sneh zero point determinations. Thc
rmHll1is should cheek within two-tenths of n scale division.
The trne zero point of l1 balance varies from day to day and
HllOUlcl be determined each time the bltlanco is used.
Exercise 2
Object.-Determination of the Sensitivity of a Balance.

Procednre.-Thc sensitivity of [t bnlance is the displacement of the zero

point proc1ueed by fL weight of 1 mg. Determine t.he true zero, or rest,
point of the blllalloe. Place the rider on the first division of the beam scale
aIllI det.ermine the new resting point Oil the index scale.

The differcnco between the two rost points gives tho sensitivity
of tlw habnee in terms of index seale divisions. Assume that the
difference in the rest points is two, then 1. mg. causes a displace-
ment of two scale divisions. Sinee the index scale can be read
to one-tenth of a division and there are 20 one-tenth divisions,
the balaneo is said to he sensitive to 0.0,1) mg.; that is, the smallest
weight which ean cnuse a readable deflect.ion is 0.05 mg.
Detormine the sensitivity of the balance with loads of 10 Om.
on o:;wh pan and with 20 Gm. on each pan. Tabula.te your
results.
WEIGHTS
Figure 8 illustrates a set of analytical weights. Analytical
weights may be purchased in graduated sets; a set in which the
26 QUANTITATIVE PHARMACEUTICAL ClIEMIS']'RY

largest weight is 50 Gm. and the smallest 5 mg. is satisfactory

for most analytical work. '1'he 1 Gm. and all larger weights
arc usually made of brass and pla.ted with gold or coated with
lacquer to prevent corrosion. The fraction:1l weights arc usually
made of platinum, tantalum, or aluminum.

FIG. S.-Set of analytical weightR.

Comparatively accurate weights can be purchased, but all

analyst must know that his weights arc aecnmtc ltnd Hhould,
therefore, always calibrate the weights at least OIlec each year.
Exercise 3
Object.-Calibration of a Set of Weights.
Procedure.-Clcan the set; of weights (,0 lw enlibmtod by wiping UWIll
thoroughly with a dean o1oj,h and hrush Ow weigh!' box with It Htij'f hrllRh.
Mark duplicate weights by punehing one 01' two dots on them so 1,hnt
they may be readily distinguiHhed. Determine thc zero point of tIl() Iml-
anee. Place the 5 mg. weight 011 tho left pan and the ridor OIL the 5 n'lJ;.
position on the right arm of the beam scale nud determino the zero point.
again. Transfer the 5 mg. weight. to the right-hnnd pan mtd pIneo tho 10
mg. weight on the left-hand pan and secure perfed balaneo by nleLtllH of Ute
rider. Proceed in like manner with the remnining woights. Always plant)
the unknown weights on the left pan and the known wnights 011 the rigllfi
pan. Tabulate the data accorciing to the following illustration:
 REMARKS AND GENERAL DIREC'l'IONS 27

CAUBllA'l'ION CHAm'

IJcft lUght Rider

rending
I Actlllli
weight
I
Aliquot Actual
part correction
_ _ _ h .

0.005 JUder +0.0050 0.0050 0.0050 "- 0.0

0.010 0.005 +0.0052 0.0102 0.0100 +0.0002
0.010 (),Ol +0.0001 0.0103 0.0100 +0.0003
0.020 0.01 + 0.01 -0.0002 0.0203 0.0200 +0.0003
0.050 0.02 -I- 0.01 -I- etc. -1-0.0042 0.0500 0.0501 +0.0001
O.10() 0.(1) + 0.02 -I- ok -1-0.004,5 0.1003 0.1002 +0.0001
0.100 CUll) + 0.02 + (lto. -1-0.0044 0.1002 0.1002 0.0000
0.200 n.IOn + 0.100 -\-0.0008 0.2013 0.2004 +0.0001)
0.500 0.200 -I- c;tc. -\-[J. 0044 0.li020 0.5010 +0.0010
1.0 O.flOn -I- etc. -1-0.0040 1.0036 1.0019 +0.0017
1.0 0.500 -I- etn. -I-O.0()24 1.0()20 1.0019 +0.0001
1.0 o .500 + etn. -1-0.0034 1.0()30 1.0019 +0.0011
2.0 1. -\- 1 -0.0014 2.0042 2.0038 +0.0004
5.n 2+1+1-1-1 0.0000 5.0128 5.0006 -1-0.0032
10.0 5+2+1-1-1-1-1 -0.0064 10.0192 10.0192 0.0000
lO.O 1)--1-2-1-1-1-1-1-1 -0.0052 10.0204 10.0192 -1-0.0012
20.n 10-1-10 -0.0024 20.0372 20.0384 -0.0012
50.0 20 -\- 10 + 10 -I- etc. -0.0032 50.0992 50.0950 -1-0.0042

The information secured in the first four columns is all tha.t

is required for a ealibmtioll of the weights in terms of relative
units. This method of ealibl'l1tion is sufficiently accurate for all
ordinary quantibtive work. The calculation of the actual
eorreetioll is found by t.aking t.he 10 Om. weight. in t.erms of the
preliminary standard and finding the aliquot part of this weight
represented by eaeh of the other weights. In our table the actual
weight of the 10 Gm. weight in terms of the preliminary standard
was found to bo 10.0192; the aliquot part of the 5 Om. weight
will then bc 10.0192/2 = 5.0096; the aliquot part of th~ 2 Gm.
weight will he 10.()192/5 = 2.0038; and the aliquot part of the
1 Gm. weight 10.()192/10 = 1.0019; etc. The aliquot part of
10.0192 is represented in the fifth column as calculated above.
In tho sixth colulnll ILIe given the actual corrections as plus or
minus depending on whether tho actual weight is greater or less
t.han t.he aliquot part.
The corrcetiolls in tenths of a milligram should be tabulated
on a card, and the card fitted tightly into the cover of the weight
blJJ;:,
28 (JUANTI'l'A'l'IVB l'IlARMACPJU'l'IG'AD CfIll.'MIS'l'llY

Weight, ( ~flI'l'e(\Li()ll, l~(Irre!"ti(}Il,

WeigllL
grams ().1 mg. O.lmg.
._-----_,----- ---- ",~------- .. ~------'-

O.OOri 0 l.O +17

0.010 +2 l.O -I- I
0.010 -I- a 1.0 'HI
0.020 +3 :J.O + '1:
0.050 0 5.0 -H12
0.100
0.100
+ 01 10.0
lO.O
0
+12
0.200 -I- \) 20.n -12
0.500 +10 50.n -1-42

Riders.---Ridel's are supplied ill fi and 10 mg. weights. AlwLLYs

eheek the weight of It new rider bei'ore weighing agaillst; the
0.005 Gm. weight.
QUestions and Problems
1. Why lllust tho qllalit:1tive tCHts pl'(!(lcc1e the I\S~lly to obtain neeurate
daLa l'cll1tive to j,he purity of lUI o!lieial snhstiLllcC'?
2. What is a GOOlilt erueiblc? Why should lltihesLOH fibcr WlLHht!d with
hyill'oehlorio neid be lI~ed in preplll'ing this cl'lwible for u,~o ill lilt 1'1\ t.io n ?
3. What is It dCiiiecatol'? Whnt two ltgents tHO uRed most; (lolllIllonly to
uehydmte ollicinl samples heforc n~sny?
4. What factors alIcet; tho sonsitiviLy of II ball1nce?
Ii. Why al'e ngnte knife edges lUld agate sllpports used in Imlalloes?
6. Why should the true zero point uf 11 Imlnllee be determined lwfOl'C
each weighing'/
7. Eyvi'JirL hc\\{ the SCllRit.ivity of 11 hnl:lIIllo Illay bo detormilled wiLh It
lond or Ir) '.'fill '''I each pitH.
 OHAPTER II
GRAVIMETRIC ANALYSIS
THEORY
'1'he student should be familiar with the modern chemical
theorics which apply in quantitative analysis from the study of
general ehemistl'Y and qualitative analysis. In quantitative
nnnlysis, theoretical considerations must be understood and
applied in order to know the full explann.tion of what takes place
in an analysis. A brief review of the fundamentals of the various
theories with their quantitative application will be given in
conjunction with those procedures to which they apply.
The reactions of quantitative analysis take place in accordance
with the established laws and theories of chemistry, i.e., theory
of ionization, law of mass [wtion, common ion effect, reversible
reactions, and solubility product principle.
Theory of Ionization.,-The theory of ionization is based on
the following postulates:
1. Eleetrolytes in solution form ions. When electrolytes are
dissolved in w:1ter, their molecules dissoei:1te or break up into
particles termed ions; thus, when hydrogen chloride gas is dis-
solved in wat~r, some of the hydrogen chloride molecules disso-
ciate or ionize to form hydrogen ions and chloride ions so that
from one molecule of hydrogen chloride, two ions are obtained.
2. Some of the ions are charged positively, others negatively;
the sum of positive and negative ch(l,l'ges alwa,ys being equal, the
solution remains electrically neutI'iLl. In the ease' of hydrogen
chlorido, the hydrog~n ions are charged positively and the
chloride ions negatively, the sum of positively charged hydrogen
ions being eqnal to tho sum of negl1tively ehl1rged chloride ions;
the solution of hydrogen chloride gas remains electrically neutn~l.
3, The properties of the ions into which the electrolyte dis-
sociates differ from the original substance. The hydrogen and
chloride iOllS have properties different from the original hydrogen
29
30 QUAN'l'ITATIVE PIIARMACIW'l'ICA.L CHEMISTRY

chloride gas, and the hydrogen and chloride iOlls differ from
hydrogen and chlorine gas, respectively.
4. The ionization of the electrolyte results in nn equilibrium.
The point [Lt which equilibrium is reaelted is dependent on the
nature of the electrolyte, the nuture of the solvent, and. upon
the dilution; thus, eloctrolytes differ in tho extent to whieh thny
ionize; some solvents, snch as ethor and bOlur,ene, lll'oduco' no
ionization, but in solvents which pl'odm:o iOllizntion, the greater
the di.lution the greater thc extent of ionization of the do(:trolyto.
A certain stl1te of equilibrium ()xists between tho ion::; and
undissoc:iated moloenlns for evcry degroo of dilution. The
cquilibrium may be ropl'es(mtml as follows: NaCl ~Nnl" + Cl--
when sodium chloride is cliHsolved in wnljnl'. Since the extent
of dissociation is dependen(; on dilution, the greater the dilution
of the solution tho more sodium ehloridn will clissoebte into iOllS
until at infinite dilution it may be reganled [LR eompktely ioniz(~d.
1'h e eqlll'l'b ., dJ 1 . [Nn,+] X [Cl--]
I num IS repl'esente )y t 10 equatIOn [Nnel] =

J( where [Na+] , [C1-], and [NaCl] rcpresent the conc()ntrl1tion

of sodium ion, chloride ion, [Lud sodium chloricle, respectively,
and J( represents the ionization cOlmtant.
Reversible Reactions.-Most of the react,iollS involved in
quantitative analysis are of the reversible type. Under eortain
conditions they may be nutdo to continue to completion, hut
und~r other conditions they may attain equilibrium before
completion, reRulting in loss of a portion of the substance being
analyzed. It, therefore, is very important to Undel'Rtnnd what
conditions must be satisfied to 1llftke the reaction go forward
to completion so that it will be of value in quanLitative work.
There are three general conditions which tend to destroy equilih-
rium and lead to complete reaction: (1) the formation of an
insoluble gas; (2) the formation of a sparingly soln ble solid;
(3) the formation of very slightly ionizecl molemlles.
According to the law of mass action, the speed of a reaetion is
proportional to the products of the molecular eoneentmtions of
the reacting substances. Since the speed of n, reaction depends
upon the concentration of cvery Sl1bstltllOe taking; pn,rt in the
reaction, the point of equilibrium will depend upon the concen-
tration of each of the components of the two opposing reactions;
 GllAVIMETRIC ANALYSIS 31
c.g., in the reaction KNO a + H 2SO, 'KHSO. + HNO s, the
speed of the reaction of potassium nitrate with sulfuric acid is
expressed by the equation:
Speed = [KNO a] X [H 2S0 4] X k where k is the affinity con-
stant of the reaction.
The speed of the op posing reaction is expressed by the equation:
Speed = [In-ISO 4] X [HNO a] X kl where lei is the affinity con-
stant of the opposing reaction. At equilibrium the speeds of
the two reactions arc equal.

::mel

Since hi [mel k are ()Ollstants, their quotient J( is a constant

termed the" equilibrium constant."
At a llefinite temperature the equilibrium constant is a fixed
v::~luc 1'01' any given ren,ction irrespective of the concentration
of the substances present;. If, therefore, the concentration of
sulfuric add is inel'eaRe(l, all of the other concentrations must
ehallgc, the eOIlccntration of KNO a must become less, and that
of both 1\.1:1S0 4 [tnd HNO a greater to maintltin the equilibrium
constant, with the result that equilibrium is forced to shift
"toward the right. In quantitative analysis, an excess of one
eornpOllent is frequcntly added to cause the reaction to go as
lW!1rly to completion as possible.
Solubility Product Principle.-The solubility product prin-
ciple is nIl application of the law of maS8 action to equilibria
whieh genemlizes the beluwior of difficultly soluble salts in their
sl1tumtcd solutions. Whenever precipitation occurs, is pre-
vented, or solution cffeetcd, the conditions to which this prin-
eiple applies arc involved. The principle may be stated as
follows: ']' he product of the concentTat'ion of the constituent ions in (L
saturated solution of a d-i:f!icultlll soluble salt for any givcn tempera-
tUTe is pmctically a constant, each concentration be'ing raised to a
power equal to the relative number of ions supplied by one molecule
of the salt upon dissociating, i.e., a difficultly soluble salt AmBn
32 QUAN'l'!'/'J11'IVJiJ PiiARMAmW~I'IOAL CHE'M!8'l'RY

upon dissociating would fumish a relative nurnber of 'In cations

and n l111iollS:
A'lnBn~ ~'1nA.+ +
nB- and, u8ing; C to dcsignaJ,(~ concentra-
tion, the solubility Pl'OdllOi; would he (C,[+)"" X (ClI-)" = solu-
bility product AmBn.
The solubility IH'oducLt; of some of the more difIieultly Rolnble
salts de:.tlt with in phnruuwcutienl annlysis aro given in the
following table:

TAllI,E I._.sOLUflILl'l'Y PUODUC'l'H ou Smm IMPOH'l'AN'l' SAW'S

r.l\l~lnpQI'I~tUl'OJ Soluuilit,y
SUL!4tnllce Ionu iTl'v()lvl~(l
C). product
---_._ .... _---_..._.
Aluminum hy<il'(/xhle., .. , .. . AJO,- X IP :l.7 X 1O- If;
Hariunl cn.rbonnte ...... ' ... . Il,L H X COi-- 8.1 X 10-'
Barium sulfatll, ....... , ... , H"H X AO., -- I.OS X lO-If)
Culduill c(Lrhflnntn ......... . Cit 1-+ X CO,"" !l.3 X lO-o
Cltleiu1l1 mmlato., ... , ..... . Ca++ X (:,,0.,-- 2.11 X lO-'
I.lmtd carbonate ............ . 18 Pu+'" X CO,-" a.a X 10'11
JJI:md !-1uIfatn ..... , . .. , ., ... . 18 Pu"" X 80.,-- 1 X 10-'
:MngncBillm fLmnlOlliulH pllOH-
ph(Lt(l ............... ,. MIIH X NIl," X 1'0,--- 2.5 X 10-1'
Mag;ll(.!~i unl hydroxich~"
..... . 18 MgH X (OIl-h 3.4 X 1(1-11
!\lngneshnu oxalate ........ , 18 Mg++ x (;,0.,-- 2.fl X HI""'
Menourie sulfid,!. .......... . 25 Hg+) X s-- <1 X 10-"
Mercurous cl1loride ......... . 2;' JIg+ X C]- :J.5 X 10-"
Silver bl'omitln ............. . 25 i\.g+ X 111'- 7.7 X lO-13
Silvcr chloride ............. . 25 Ag+ X Cl- 1.f> X IO'D
Silver iodide ........ , .... .. . 25 Ag+ X I- O.!l X 10- 10
Silver thiocytl,llfLtlL .. , ...... 25 Ag+ X SCN- 1.2 X 1O-li

When a number of different kinds of ions are present in tho

same solution, the gron,test C:Olloontmtioll that imy Olie of them
can attain is determined by the others. Thus, no grent con-
centration of Ag+ can be present in a solution in the presence of
C1-, for the two ions unite to form a pl'o()ipitate of tho difficultly
soluble salt, silver chloride. Silver chlol'ide is slightly soluble
in water, the solubility being about O.OOOOl mole (1.5 mg.) pol'
liter. When this solubility is exceeded, a procipittttc of AgOl
forms which is in equilibrium with the dissolved silver chloride.
When the equilibrium is established, the supernatant liquid is a
saturated solution, and the tencleney of the solid to go into
solution is exactly equal to the tendency of the dissolved salt
 GRA VIMliJTlUC ANALYSIS 33

to precipitato. When this condition exists, the following scheme

of equilibria is set up:
AgCI~ AgCl )Ag+ + Cl-
solid dissolved dissolve<l
ppt. unionized ionized
The ionization equilibrium expressed according to theory is
(Ag+] X [CI-] ionized T?'
-~~--"--~---~' '--,--'~.- - = .n. = 101llzatlOll constant
[AgCl] umolllzed
Since the solution is S:1turated with AgCl at a given tempera-
turc [weI the COlwcntl'ation of unionized (AgCl] remains constant,
it follows that the product J( X [AgCl] also remains constant
and that in (J, saturated solution of a d'i;Uic'ultly soluble salt, the
lJl'o(iuct oj the concentrations of 1:ts ions is constant.
In the calculation of the solubility product, it is customary to
express the coneentmtion in terms of moles pCI' liter. A saturated
solution of silver chloride contains 1.5 X 10- 3 Gm. (0.0015 Gm.)
per liter. The molecular weight of silver chloride is 143.34.
In terms of molarity, therefore, the solution contains
1.5 X 10- 3 -i}
143.3-4- = 1.1 X 10

moles per liter. At this dilution t,he dissolved silver chloride may
be assullled to be cornpletely ionized, AgCl~Ag+ Cl- so+
that each mole of silver chloride furnishes 1 .mole of silver ions
and 1 mole of chloride ions. The ,solubility product, S.P., then
[Ag+] X [Cl-] = (1.1 X 10-5) X (1.1 X 10-) = 1.2 X 10- 1,
and the iouic product is equal to the solubility product.
If tho solnbility of [1 compound is known, the solubility product
may be ealeuhttcd; c,g., a saturated solution of silver iodide at
25C. (]ontains about 0.00235 mg. of AgI per liter. The molar
solubility is equnl to the solubility in grams divided by the
gram-molecular weight, or
0.0~~~~823~ = 0.00000001 = 10-&.

Assuming 100 pel' cent ionization at this dilution', the concentra-

tions of Ag+ and of 1- also equal 10-8, since each molecule of
34 QUAN7'ITATIVE PHARMACEUTICAl, CIIEM1STilY

AgI forms 1 Ag+ ion and 1 1- ion, and the solubility product is
equal to the ionic product;
GAIt+ X C1- = 10-8 X 10--8 = 10-16 = S.P. of AgI
From the above calculation of solubility produo!;, it enn he
predicted that silver iodide will precipitate if CAIt+ X (Jr-lHJeoInPs
greater than 10- tIl ; that in fI, supersaturated sollltiou OA,,+ X Or-
becomes greater than IO--ltl; tha!; in it satura(;ed solution OA,,+
X Cc will be equa'! to 10- 16 ; and tIm!; to dissolve procipitatod
AgI, OA"+ X G1- must be less than 10- 11).
In the above illustration, the soluLili!;y pro duet of silver iodide
was calculated from solubility data. COllvun,;ely, if the solubility
produ(Jt is known, the solubility of a (Jolllpound m:w he <Jalcu-
lntecl. In the table, the solubility product of silver chloride is
given as 1.5 X 1O"-ll1 fLt 25C. How much silvor chloridu will
dissolve in IOO ceo of wator at the above temperature?
S,P' AltOI = (JAg+ X OCI- = 1.5 X 10... 10
If CAg+ = x, theu OCl- = x, sinee CA .+ = CC1-
a;2 = 1..5 X 10- 10
X = 1.22 X 10-0
:c is expressed in terms of moles per liter. Since the soluuility
of AgOl is equal to 1.22 X 10- 5 moles per liter, the solubility
1.22 X 10-0
in grams per 100 ec. is equal to ---fo--- tImes the grnm-
molecular weight of Agel or 1.22 X 10-- 0 X 143.5 = 0.000175
Gm. Agel soluble in 100 cc. of water.
The solubility produet is thus seen to be an ultim:1te value
attained by the ionie product when eCluilibriuIll hits beon- estab-
lished between the undissolved solid and tho diffienltly solnble
salt in solution. If the product of the oonconiimtiou of ally
pair of ions in solution is made to exeeed in vahw the solubility
produet of the compound formed by their union, preeipitation of
the compound will take plnce until the pl'ocluet of the ionic
concentration is exactly equal to the solubility product value,
and when the procluet of the ionie eoneentrations is made less
than the solubility product value, the compound formecI by
their union will dissolve until the pro duet of ionic concentrations
is equal to the solubility product value.
 GRAVI1I1E1'RIC ANALYSIS 35
Common Ion Effect.-The equilibrium constant does not
change no matter what the concentration of the reacting sub-
stances may be. The relative concentration of the reacting
substl1llCeS 11.1:1Y change but there is no change in the equilibrium
constant. When two substances furnish an ion in common so
that the concentrations of positive and negative ions of an elec-
trolyte are unequal, the law of mass action causes equilibrium
to be maintained; thus, when a solution of silver nitrate is added
to a. solution of sodium chloride, the chloride ion is mdlnentarily
present in a concentration such that its ionic product with the
silver ion exceeds the solubility product of silver chloride, and the
insoluble silver c:hlol'ide is precipitated:
Ag+ + CI-->AgCl t
'Vhcn an equivalent amount of silver nitrate has been added,
and the system has acquired equilibrium, the concentration of
silver ions will be exactly equal to the concentration of chloride
ions.
If to the supernatant liquid which is a saturated solution of
silver chloride a small amount of a soluble silver salt or a soluble
chloridc bc added, a slight further precipitation will take place.
It follows from the application of the equilibrium representing the
ioniztttion constant
[Ag+J X [Cl-l = ](
----------
[AgCI]
that if the concentration of silvcr ion be increased by the addition
of a soluble silvcr salt, the conc:entl'ation of chloride ion must
decrease and, conversely, that if the concentration of chloride
ion be increased by adding a soluble chloride, the concentration
of silver ion must decrease since thcir product rcmains constant.
This decrease in the concentration of the ions in either ease call
be accomplishod only by the union of silver and chloride ions to
form insolublo silver chloride forcing the reaction toward
completion.
The (lommon ion efient is used frequently in gravimetric
pharmaceutical analysis to drive reactions toward completion.
Other examples of common ion effect will be discussed in conj une,
tion with the assays in which they occur.
36 QUAN~L'I'1'A'1'IVE PllARMACEU'l'ICAL ClIltJMI8:I'RY

Questions Ilnd Prohlems

1. What !lrc j,hn posi,ullltes upon whidl thn j,\H)OI'Y of ionization iH hmlccl?
2. Whttt i8 !t revCl'Hil,\c l'uaetioJl '/
3. What eOlulitious 1,eIH\ to <1eH1.roy C'ClIliiihl'illlll [lm\ o:mHll tt I'Ullc:tioll 1.0
go to oomp\el,iou?
4. Define mu\ illuHtmtc 1he so ill biHty p!'oduct pril)(:iple.
6. O.2't7 mg. of barium Hul[a1,o diHRlllvo ill 100 on. of watul' at 25"C.
ExprcRH tlw solubiJit;y ill torwH of IlIolal' };olllbility.
6. From the Rolubili1.y of 1Ia80 4 givCJn illlll'olliem 5 cn,lcailate ilK Ho!uhility
lll'oduc:t assuming 100 pel' cour, ioniy,lltioli.
7. Tho solubility produu1; of unlcilllll .~lIlfntn i:, ltppl'Oxillllttuly 2 X :JO-1.
Cf1}eu]:tto i,lw aJIlOIlJ\ t of mtloiulll HlIJI'ntn Noluble ill 500 un. oj' wal,(!'. A BHllflIC.
100 per (\[1111; ionization.
S. Illustrate how l\OlHlllOn ion effcet HUty be utilbled to HOC\Il'O (jutllltitativl'
pl'ecipil.atioll.
9. A sntuwtcd solutioll of lead chloride ttl 20C. contl1ills 0.07 moJn per
lit~l'. This solution is 8() lll!r (:enl. ionizell. Ca\culate the !'loJlIhilij,y lll'l)(luet,
 CHAI)TER III

GRAVIMETRIC METHODS
Gravimetric analysis implies that the substance to be deter-
milled is to he separated frum a weighed sample in the form of a
compound of known cOlupositiou and vieigltccl. I"i:uowillg the
weight of the original :,atnplc and that of the product, the weigbt
and percentage of any eomponcnt common to both can be ca1-
culatocl. The product to be weighed in plu\'l'lllacellticnl analysis
may be obtained by anyone of vfLrimul methods: (1) It may be
pmcipitatecl from Holution; (2) it may be the decomposition
pro<ineli resulting from ignition of a compollnd; (3) it may be
deposited on an dnelrode by eleetrolysis; (4) it may be separated
from other substnlwes by exLradJioll "with a solvent; and (5) it
may he obta,inOfI by absorbing; [1 gas in some substance of known
weight. and findillg; t\tn increase in weight produeecl by the absorp-
tion of tlw gaH. Tlw firHt two methodfl eomprise the subject
matter considel'o(l in this (~hallt.(]l'.

Exercise 4
Determination of Chloride Ion in a Soluble Chloride .. --Chloride
ion is deturmillcd gmvillleLrically by preeipitating and weighing it
ttH Hilver eldoride. An excess of solution of silvCl' nitrate, slightly
acidified with nitric add, is added to the solution of soluble ehlo-
rich). 'fho preeipitate is tiHerocl out, wf1shecl, dried, and weighed
as silv(~r c:ltloJ'ic[n. Otlwl' 8ubstnlH:us whidl yield insoluhle silver
snlts lllLlst bn nhsunt.
No llldJlOtl for i,he gl'[willwtl'ie determination of c:hloride ion is
ofIieial, bllt the pl'oemlure constitutes [1 elassie example of gravi-
metrie [1wLlytieal technique with which every student should be
familiu,r.
Object.--Assuy of Sodium Chloride.
Materials Required.-O.25 Gm. of sodium chloride.
5 pel' cent ~ilvcr llitmtc solution.
37
38 QUAWI'I'l'A'I'IVB PIlAUM11CEU']'ICA.D (!JIEMIS'l'IlY

DilllLeclliiLric [Leicl.
As]wstos IilJCI'.
Procedul"e.-l. Acoumtely.woigh two Harnph:s of 0.2 Lo (l.a Gm. of Hodillm
ehloriclc.

This is most easily done by weighing the slLmp]e tube (weighing

bottle) and contents nccurnLely to III mg., without touehing it,
entering Lhe weight in the notehook. Holel the weighing bottle
dil'ccUy above it 300 to 400 ec. heakm, labeled I, remove the
stopper, [tlld carefnlly trnIlsfor 0.2 to o.a Om. of the sample to
the beaker. I{,()piaoe tho stopper and agltin weigh the weighing
bottle to 0.1 Illg., entering the weight in the notebook. The
first weight le:.;s the seeond i::; the weight of sample taken. In
like manner weigh n second 0.2 to O.~i Gm. sample into bea1cer II,
cntering the weights as before. Treat eaeh sample as follows:
2. Completely diRsolvo UIO Hnmple in 100 to 125 (:e. of dist.illed water.
Add ahollt. 1 co. of (liIllted niLrie Iwid slowly with stirring. Test the solution
by touehing It !;trip of hlue litlllUH paper wiLh tho moist sUrring rod. If it
is not aeid in remution, add Huftic:ient ac:id to lllllko it ~O. Measure out 5 CCI.
in exceHS of t.he Hmount of silvor nitmte solution UlI)orctieally required to
preeipitltt.O all of the chlorine [Ii! silvor ehloricle. Add the silver nitrate
solution ill smnll Hllc(:essive port,iolls, st.irring constantly with a suitable
glass rod. Covet' the bpaker with a watch glass, and, with occasional
stirring, gl'ltchmlly heat tbo mix Lure to the boilillg point. Tum down the
fll1me [1Jld digost the mixturo, without, boiling for 10 min. Thii:l trontment
eongllbl,es tha preeipita.tu, oauses it to soUle, lULU leaves tho sup~rnatant.
liquiu clear. Add 1 or 2 drops of silvcr nitrate ~oluLion to the hot super-
Imt!tnt liquitl to teHt for eomplotc precipitatioll. If n precipitate forms, add
5 ceo more Hilver nit.n1to solut.ion, stir, allow to settle, and test ItS before.
Set the beakers aside away from direct sunlight to allow the precipitate to
settle.

The solution is acidified with nitric :wid to prevent the precipi-

tation of' other substances insoluble in water bui; soluhln in nitric
acid sueh as carbonates, oxidns, and phosphlttes. The twid ltlso
helps to coagulate any colloidal silver chloride. ']'00 gl'cai; an
exeess of nitric acid must not be added, since iI; exerts SOIlle sol-
vent aetion on silver halides. The uddified solution should not
be heated until the silver nitrate has been added, since chlorine
may be liberated and lost, thus:

NaCl + HNOa-~IICl + NaNO a

5HCl + 2HNO a--,>3C1 + 4H 0 + 2NO
2 2
 GRAVIMETIUC ME'I'HODS 39
The silver nitrate is added in excess to drive the reaction to
completion through the production of common ion effect, assuring
complete precipitation:
AgNO a + NaCl~AgCll + NaN0 3

The precipitation must be performed away from strong light,

since silver chloride is decomposed by sunlight with loss of
chlorine, while in uiffllsed light the enol' from this somce is
negligible.
The preeipitate is digested near the boiling point to coagulate
colloidal silver chloride whieh, if present, would pass through the
filter.
3. During IntcrvalH in the above procedure, carefully dean llnd prepare
two Gooeh (It'llcibks for filtration as l)l'evionsly deRl~ribl\d. After the ern-
ciblml luwe boon pl'Oporly prepared, heated to eom;tnnt weight, and weighed,
lit them into tho suet,ion flask. Be sure that the erueiblos can he idlmtified
as I and II l)y !L HlIitable mark. Dccnnt, most of the supernattmt liquid
into tho Gooeh crueihlo, lI~ing genUe SUC[,jOll to ha~ten filtration, guilliJlg the
liquid into the r:rucihle by menns of a glass rod held n,gllillHt the lip of tho
bet1kcl'. WtUlh Lho pl'eeipitate Ulree times with 15 ce. portions of hot, wlLsh
solution prepared by adding a chop or two of dilute uitl'ie ILcid to 200 ee. of
distilled water.

In pouring the clear liquid through the filter, retain as much of

the precipitate as pmlsible in the heaker. When the third wash
solution has been added to the precipitate, transfer the precipitate
into the Gooch crucible using a stream of hot wash solution from
the wash bottle and a "policeman" to dislodge all particles which
adhere to the sides of the beaker and guide rod. vVash the
precipitate on the filter ,vith severnl small portions of acidulated
wash solution, allowillg ench portion to ruIl through before ndding
anothe!'. Continue washing the precipitate in this manner until
1 to 2 ceo of filtrate give no test for silver upon the addition of a
few drops of diluto hydrochloric acid solution. When tested in
this way, the filtmte should give no greater turbidity than a
portion of the wash solution l)eing used tested in the same manner.
4. When the filtrate is shown by the test to be free of silver nitrate, wash
the Pl'()eipitate twitle with 5 ~C. portions of distilled water to remove most
of the nitric acid ret:tinerl by the precipitate from the former wash solution.
Apply strong suction to remove as much liquid front the precipittLtc as
possibill. Dry the crucible to constant ,weight at 110 to 120 0 in an air bath
40 (JUAN'l'I'l'A'l'lVliJ l'HAlUli[AGIW~l'IC'JlL L'l1/~MIS'l'Ul'

01' elcetl'ic oven until SUC(~eBE;ive weir~hillg;,9 tliffm' uy

no mO),(1 Lhan 0.0002
Om. Allow Lhe cl'lleibll!~ to eool ill [), (lc'Ric:eaj,ol' bnfol'e wt.,i!;hillg. The
weight, of the cl'Ueible :l1ld p!'eeipitnte miIlllH tho weight, of '!.he el'llcihle giveR
the weigllL of Hilv(q' ehlol'i(le obtained frolll tho Rlllllpl(\.

The pn,eipitat(\ is wru,lwtl to r()Il1OV(~ HolulJ]e HaHN 1)J.'oscmL ill the

solution, ehiofly sodium lliLr:tl,o :L1l(l exee~R SilV(1]' nitrate, Hot.
WIlRh solution in(lrense::; tlte fmlubiliLy of thm~e mt!(,fj ml(l keepN
the silv()r ebloricle hom pns::;ing into i,hn (lolloillnl enndil,.ion. The
solubility of sil VOl' ehlori(le illel'enSCs slightly in hut solut.ioll, hut
this is compensated f(j]' by tho slflullm' amouut of WltRlt sulution
requin1(1.
Silver e111ol'idn iN tLppl'ueirLhly vobtil(1 whon ignitnd, so if,
should be dricclat. a. rela.tively low tnH1[lemturn,
The rnaeti()ll illvolved ill tJIO eaklllation (,1' t.h(~ :t.IllJnmt of Nilv('l'
nitrato solution to be ([Red :I.1Il1 of tho pUl'ity uf j,I10 FHuliple is
AgNO;! + NaCl->AgCl +. NrLNO;!
1(j~1.8~) 118,4rj 14:~.:a S:i.Ol

BoU,ln hero]'" l'()Jll[Jv:Ll o[ i'nlllp]l'. , ]o1.,:3I1S0 I oJ, ()o!:~O

Bot/,I() nfL!1l' )'1'IIlDv:ll of ,~;!llIJlI, ,. H,()'I:21l l:l.Tllii
Sam pIe, , . , . , .... , ... , .. , . , , ' . , ' O.:Hon (),:lI!)[J
Cl'uc:ihlo .. , .. ,. ,."."',",, .... IS.21(it) I'i ,f{S:!.7
Cl'llI:ible nnc! "iIv(!]' I:lthll'i,l" '("1,, . , I.n,()~W1 IX, (\40H
CI'Il!1ih10 nnel Hilvol' I'Itlol'idn 2,1"., 10.0:.l0l Ig,('"tn;{
(;I.'III,,!ll](: nnd silv(i]' eli loddo ad , ' , 'I/!)' O:W() liUHO:.l
(,lllllhll, .. " . , ... " " . " " " . " , IS,~l'[(',~\ "7,gS~'{
Silver dll()ridr~ .. , ...... ,., ".,. O.77:\:~ O.'lG7,rj
-----<-__._-------.~-.-------~--~-.------

Tlw equn.j,ion shows that 1 ntoleC:l1lu or Ro(lilllll chloride l'(){wtH

with 1 llloleeule of silver nitrate, Fl'om tlw lllole(lu!n.r \Voight"
N ael = 58.45 :.mel AgNO:1 = lOn.S!), t.hen'.i'orn, tlw LhoOl'nti('al
amount of silver uitmtn solution req uiJ'(~d 1.0 pl'ndpit:Lie n,ll of
the chloride ion as silvol' ehlOl'ide can. be ealnubtecl [1,,'1 fOUOWH:
169.89/58.45 = 2.!)O(i(j lim. AI};NO;J required to pl'o(iir>ik~tn tho
ehlol'ine from 10m, of Nnel. Sin<:(', from the elata, 0.31()0
Gm, of NaCl were llflCll, then f),SHiO X 2.9()(\() ,= O.918!J Gm .
AgN0 3 required, Sinee the l'eag;rmt silver nitrate is It 5 p(~r
 UUA Vl.ME:1'IUC M1E'l'EWDS 4r
cent solution, oneh cuLic centirnr,ter must contain 0.05 Om. of
AgNO a and the amollnt of solution required is 0.9185/0.05 =
18.:37 ce. of [) pur (~tmt Kulutioll.
Tlw wfligltt of thti K~LI)J pI<: and the weight of silver chloride
oht:Lincd front iL }>nillp; ktlown, the pel" cent purity of the sample
01' t1w weight of dtl.ol'illti in the Rample may readily be cnleul:1ted:
5gA5/U;i.;{4 = OA07S Gm. NaCI cquivf1Jent to 10m. of
Agel. RiJl(:e IOn!. or Agel if! eqllivlti0nt to 0.,1078 Gm. of NaCI,
then 0.77:32 Gill. o[ AgC:l i'oulHl i,; cqnivalent to 0.'1078 X 0.7732
= O.:s15:~ elm. Nael. The pOl' cent purity of the sample iK
o.:n[j:~ I()) . /""'.
t IJere f oru UjT(it) X . (
= 9H.' 8 pel' cellt.
Having etdeubtu( I tIl() equivalent weight, the mlloulation of
pel' (:ent pmity lllay J)() ubtnirwc\ by a single formula applicable
in all graviwntri(: d(~(,(:],]Jljllati()ni-i, Chus:
WXE .
--I<:r--- X 100 = 1W1" epJ)t whpl'C' TV is weight of AgCl, E is
tho eqllivalollt weight, und S tho weight of sample; ('.g.,
0.77:12 X OA07S ,..
. ii:3Iif6 - - X 100 = 99.,8 per (lont.

1'0 enJeuhte tIle per t:Cillt chlorine in the sample, substitute

the weight of chlorine, ;::5.45, for that uf NnCl.
:j5,45jt4:U14 = 0.217<1 Gm. CI equivalent to 1 Gm. of AgCl.
SubsLitnting this valun for E in the above formula gives the per
eent ehlorbw in the s:tlllplo.
The quantity hy woight of an element or compound which is
equivalent to Olle 1m!'l; by weight of o;ome other substance is
termed a c}w't/l,'il:al facto)'. For example, the ratio or factor
NaCI/AgCl = liH.45/14:3.B4 = 0.4()'18 means that 0.4078 Gm. of'
puro NaCl eOl'l't!sponds to 1 Gm. of AgCl. To find the amount
of Nnel relll'f)Rentecl by [Lny given weight of Agel, therefore,
tho weight of AgCn obtained is multiplied by this factor. Thus
0.1 Om. of Agel iN equivalent to 0.1 X 0.4078 = 0.0408 Gm.
NaCl. Likewise, Ag/ Agel = 107.88/143.34 = 0.7526. This
menns thnt 10m. of Agel contains O.752() Gm. of silver and that
if we' wish to C!1Ieuln,t.e the weight of silver in a specific weight of
silver chloride, the latter need only be multiplied by this factor;
e,g'l 5.2 Om. of AgCI contain 5,2 X 0.7526 = 3.9136 Gm. Ag.
42 QUAN'1'I'1'A'1'IVE PHARMAOEU'1'J(JAL CIWMIS'1'RY

The gravimotric method for the dctenninntion of chloride ion

ll1ay be applied to estimate the 111ll'ity of most soluble ehlol'ides,
brornilles, iodides, eyanirles, thioeyunaLcs, cte., us wdl as to the
si;rmdurdiz[ttion of hydroehlorie twid solutiolll'l and the estimation
of silver eomp0111H.ls. ChlorateR, pCl'cblomtes, and other oxy-
adds of the halogens may be lletennined by this methocl after
reducing them to the halides. The vol uIllctric precipitation
Jlwthod has superseded the gmvimetl'ie method because it can
be carried. out more mpidly. Tho gl'et1test objectioll to this
method is that impurities in the form of Imlogell salts are included
in the determ.inatioll, the results being in reality a measure of t}w
halogens prescnt.

Questions and Problems

1. Why should the solution be aeicInlflted WiUl nitric acid?
2. Why add Hilver Ilitrate ill oxeOHS in t.ho procipitlltion of tho c\hloriclc?
3. Why is the precipitate w!\Hhcu with waLor Hlighlly a(~idulated with
n itl'ic acid?
4. Why iR it best to wltHh the preeilliLl1te by dCetlllttltiOll ]'elfol'o it; is
transferred to the Gcweh cl'Iwible't
6. Why should this dctcrmiwtLioll he earricd onl, away from direet SllIl-
light?
6. Name several oflieial suhstallces 0 Lller Llllm sodiulll ehloride w llioh nUty
be assayed by an analogous lllethod.
7. What snbstnnceH, if pl'esout, would interfere with the deterllliu!ltioll
of the purity of n soluhlc chloride?
8. In the preeipitntion of chloride fwm 0.(i00 Om. of !11l unknowll 811b-
Htl\I\Ce, [I. precipitate of O.(i800 Gm. of Agel WIUI obtained. CaleulltLe tlH~
pur cent chlorine in the unknown.
9. How many grams of AgN0 3 IIl'llIH1CflHH[\ry io preeipiLate the c:hlOl'illfl
from 10m. of 50 per cent zine chloride solutiol1'l
10. When call it be nssuIIled that 11 erL1cihle iH of eOllHj,[mL weight'l
11. Why are samples usulilly dried before POl'l.iOIlH llrc weighed for
nIlalysis?
12. Give a mtioual pro{l(!dure for the dcj.cll'IniulItioll of Hilver by the
gravimetric method.
13. What is meant by the terms chemical factor :.Lml ch(!mical CCjlti'Vlllcnl?
Illustmte.

Exercise 5
Determination of Sulfate Ion in a Soluble Sulfate.-The
sulfate ion in a soluble sulfate may be determined gravimet,rically
 GRA VIMETRIC METHODS 43

by precipitation as barium sulfa,te, the precipitate being collected,

dried, ignited, and weighed.
Object.-Assay of Sodium Sulfate.
Materials Reqllired.-l Gm. of sodium sulfate.
Barium chloridc T.S. (12 pCI' cent).
Procedttre.-Weigh aceuraLcly about 0.4 Gm. of the dried salt, obtained
by drying about 1. GIll. to eonstmtl, weight at 120 D C., dissolve it in 200 CC.
of distilled W!Lter, and ad(I 1. cc. of hydrochloric acid. Heat to boilillg, nnd
gradually add an exeeSH of hot barium chloride T.S. Heat the mixture for
30 min. OIl a water bath, (Jollnet the precipitate of barium sulfate OIl a filter,
wl1Hh it until free from chloride, dry, igl1ii,e, lmel weigh. The weight of the
barium sUlfltte thus obtained, multiplied by O.GOSu, indicl1tes its equivnlent
of Nl12S04.
The solution of the sulfttte is aciduhtcd with hydrochloric
add to prevent the preeipitation of carbonates, etc., which are
soluble in hydroehloric acid solution. The acid also increases
the solubility of bn,rium sulfate slightly and therefore promotes
the growth of large crystals, since small crystals dissolve more
rapidly than large ones. The small crystals expose more surface
area per unit of weight than do large ones. Consequently, they
dissolve more rapidly, and in a saturated solution tIde larger
crystals with small surface area exposed grow more rapidly than
they dissolve. The solution is heated to boiling to expel elis
solved CO 2 An excess of hot barium chloride solution is added
to insure complete precipitation and to decrease the solubility
of barium sulfate by common ion effect. The mixture is digested
011 a wate)" bath for 30 min. to allow the larger crystals to grow
at the expeu:-Je of the smaller ones., since the latter are more
rapidly soluble.
The bl1rium sulfate is directed to be washed on a filter to
remove exeess BaCh and the NaCI formed, but the washing
process may hc earried on t more rapidly by washing once by
decantation. The precipitate is then transferred quantitatively
onto an n.shless filter by the same technique used in Exercise 4
and washed free of chlorides as shown by tests made with silver
nitrate solution on 2 cc. portions of the filtrate. Always acidu-
late the filtrate with a drop of nitric acid in carrying out the test
for chlorides. Sometimes fine-grained crystalline precipitates
will pass through the filter. This may be observed by imparting
a rotatory motion to the filtrate, causing any crystals which
44 QUAN'1'1'1'L1'1'TVE PllAllJl:L1UI!JU'l'ICMJ CIlEM18'l'llY

passed t.hrough to g:tL]wi in t.he (~cllter of the bonkcr. If sueh

are d(~teet.ed! refilter.
Barium. sulfrLte exhibits the propDrty of "dra!1;~in~ down"
other 80111010 c1111tH when it is )1reeipii,ntcd. 'l'his property il'1
known as CO-]1Tp-dlJ'z'totion. TIi.o oedlllled or co-prneipikIJ.()cl
salt eallIloi; be l'lmlOVed by onlinnl'Y washing. The en'OJ: frolll
this Romee is redllce<.l lf1l'g(~ly by prc(:illitntioll frOll] hut dilll(,o
solutions and l)y adding tho pl'()(]ipitatillg l'nagmd, Hlovdy with
continnous Htirl'illg. The phenomelloll of eO-lu'n<:ipitntion iH
particubrly liable to oecur when Imrillll1 slIlfate is precipitated

TA]l'Ll~ lI.-OFliICIAT, f4UTlWl',\N('Ji:f; ASKAYKD I,Y GltAVI~UJ'.I'ltl(' l'ltlilCIJ'['I',\'l'l()N'

A~ RULl,1Nl:l;1
--------------,---.,.-_.-_"...'-------
Woig;llt,
SulmtullC[J of kllilion l' t ()" I '
Hil.lllple, re~idul 'ae ,Ill' {hem l"(~qIL1],(!JlHmt" tiP!" emlt
Om.
_"_'_"--"--'-'" . -
_ ...... ... _..._... ........ _...... --.... ~

U.S.P.
Len.Li subaoetntn, n ......... . 1.0 Phi')(l., I'h ._ 70 1.(17:':
Sodium Bullate ....... " , . , .. 0.1 :nItSO., n.rms!; Nn,S() , ._. !Hi
Sulfur oiutnlOuL. ........... . n.r. JlaHO., tl.l:!7a H I:Ui til 1(\.r,
;:-;l

Sulfur (all IOl'm.) , .... , , . ' . , . 1.0 I1"SO., O.lii73 8 ~

\lU.r.i
N.F.
Ichth:tllUnol for (NI-L,);i'O., .. . 1.0 BaR()., O.linlll (Nl1 . ),SO., .. not more thuu 8
IchthnmmllI for total sulfur .. . 1.'; ]laRO" n.lanS ... 1fl
PotuHsimn guninenl fiUlf()ll:tU' l.fI HaS n ., 1.O::i7t,7 e,,1 [".0 1r.nCIl"RO"K (l7 :::;l

Potflssilnl1 I:mlfatn, ... , .... , . 11,0 llaNO., ().71O,; 1\,:-\0., - on

Sodium sulf:Ltl.! l!x~~i(~cated. , , '/ O.r. lIaSO I I).nml!) N""R(l, = !I!I

R. = ruagent,

in the presence of chlorai,Cls, nitmt;oR, alld tlw fm]t,c; of iroll,

alulllimun, chromium, a.nd other t,d Val(:llL rlmncm 1.:'1.
The pl'Cleipitatc may be (lric'(l and. ignited by the wet method aN
cleseribed uncleI' Genoml OI)el'uLio]1s (pal!;e 2C). The eqllation

Na~SO'1 -+ BfLCI2.2ILO-~nn.sOI + 2NaCl + 2H~O

142.05 283.42
expresses the stoichiometric relationship 011 which the enJellla-
tions are based:

;~~:~~ = 0.6086 Om. Na 2 S04 equivalellt to 10m. of BnS01

 OlViVIME'I'JUC ME'l'IIOVS 45
The gravimetric determination of sulfate ion as barium sulfate
is use!l extensively in plumnaeeutimLI analysis. It may be
nppliud to the estiuw,tioll of me/;als whiell form illsoluble sul-
fatns as b[LrilllH and lead. Sulfides and sulfur, free, and in
orgn,nie combinat,ioll, may be oxidized to sulfate and subse-
quently pl'(leipitni;()d as harium sulfate. Other compounds as
thiuslllf:L1.eS, sllUiLciH, pnl'Slilfate::;, alknJoidnJ :sulfates, ete., may
be esLim:L1.ecl by the appii()tLtioll of similar proocdurcs.

Questions and Problems

1. Why if; 1110 ~'(Jll1Li()H (If ,:oluhlc ~ulfat(J :tcidubl;cd with hydrochloric
ndd?
2. Why i~ i LII('I'(~HHa,l''y to dig;l~Hj, the prollipiLflte of Imriutll sulfate pl'cvious
to filLrn.Lioll?
3. Why Nl,ollld thu pl'u(:ipihd.ioll of Hult'a:('es bl: !O:ul'icd oul; in IlOt, dilute
solutioll '(
4. An ullkllown flmnpl(] of :1 :,:oluhlo sulfnto \V~ig;hiIlg 1.8000 Gm: yielded
O. nono elm. of nnSO.). Caleuln I,e tllO pOl'crmtnge of fJulfur in tho unknown.
15. C:t.1(:lllnLu lIt[) equivaleu L :l.t1l0Ullts of uaeh lIud the pC1'(:elltage purity
if Ow 11ll1mowIl ill Pl'O]ljlllll <1 \\'()J'(: MgSO." 1\:280." Ale(SO.,)" or H 2SO,.
G. 'Writo L]w eqllfLtiO]lf1 WllrCN(]ltl.illg tho l'()lLotions that t.ake place in the
[IS,;lI,Y of Hulflll' uilll.lllfln L.
7. How lllllr:h 1'.:11')0, wonld lin formed from 1.2000 Gm. of NfL,SOol.lOH 20?
8. Wha.t is the por cput of 80., in l\ Hample of 1(,80 4 if 0.5000 Gm, yields
UpOIJ l'uHci,iolJ wi!.h The1, t1 Vl'ccipil;ato weighillg 0,5850 (1m.?
9. How is ICltLl dotel'miuod as slllfat.n? Elee roagont load 8ubacotate,
U.S,P.

Exercise ()
Determination of the Mercury Content of a Mercuric Salt.--
The mOl'l:nry is preoipitatod as sulfide, washed, dried, and
weighed.
Object.---Asrmy of l'vlereul'ie Chloride.
Materials Reql1i1'ed.-O.5 Om. of mercuric chloride.
1 eo. of hyclroehlol'lo twid.
About 70 ec. of alcohol.
About. tl0 cc, of (:l1rbOll t.eLmchlorldc.
Procedure.-l. "Dry IIbout 0.5 Gm. of Mercury Bichloride to constant
weight. over sulfurio g,cid, wnigh IlcoUl'nt.c1y, !111(l di~solve in 300 oe. of \VarlU
distilled waLor to which 1 eo. of hydl'Ochloric [wid has been added. Pass
hydrogen sulfide throngh the cold solution until the precipitate of mercuric
sulfide roadily Bubsides, leaving 11 olear, Buper Il at11nt liquid."
46 QUAN'l'ITA'l'[VE PIIARMACEU'J'ICAL CllEMI8'J'RY

The hydrogen sulfide gas used in the precipitation should be

washed by bubbling it through a gas-washing bottle containing
water. The glass delivery tube which passes to the bottom of
the precipit:1tioll belLkel' should be att[Lehccl to the gas-washing
bottle by Ineans of a rubber tube in snch a manner that it can
be removed readily and held over the preeipittttion beaker to
wash any adhering precipitate into the JJeaker.
The hydrogen sulfide rends with the mercuric chloride to form
a double salt of mercuric sulfide lmel mel'enric chloride at first and
then this double snlt is decomposed with the fonnntion of black
mercuric sulfide as more hydrogen sulfide is passed into tho mix-
ture. Since mercuric sulfide is insoluble in adds, hydrogen
sulfide muy be employed as the pl'ceipitnting agent Hnd hydro-
chloric [tcid mt\y be t1clded to repress tho coneentl'u,tion of sulfide
ion so that mctals which form sulfides more soluble than mercuric
sulfide and which m~ty be present as impurities will Hot be
precipitated.

2. "Collect the precipitate on couIltcrpOi8()d filterA or in n, tared Gooch

crucible, wash it well with cold distilled water, !Iud [innlly with throe porj,ions
of about 10 ce. elLeh of alcohol. ThOll dose the tip of the funnd 01' of the
Gooch crucible holdcr with !L cork stopper, add sllllicicnt carboll totraehlol'ide
to covel' the prccipihtte, covel' the funnel or crueiblc with It Wl1te,h gh\t;tl, :11111
allow it to stand for about half an hour. Dl'l,in off the ~olvnllt, and w(lHh
the precipitate with further POl'tiOll8 of carbon tctmehlol'ido uutil, aftor
evapol'!lting about 1 ce. of the filtn,te, no visible residue rOHlI,iIlS. Hemove
the adhering C!lrbOIl tet1'!lchloride by washing Lhe prceipiLatc with s(wcl'!d
portions of 10 ce. each of alcohol, and, after drying ill t.he !til', transfer to all
oven and dry to eOIlst!lnt weight at about !lOoe. The weight of mercuric
sulfide, multiplied by l.Hl7, indicates its equivalent in HgCh,"

The precipitate is washed with wuter to remove soluble sulfides,

then with alcohol to remove the water. The carbon tetra-
chloride, which is added later and is ilnmiscible with water but
miscible with alcohol, can permeate the precipitate and dissolve
any sulfur formed by the decomposition of hydrogen sulfide.
The precipitate is allowed to stand eovered with earbon tetra-
chloride for Y2 hr. because sulfur is slowly soluble in earbOll
tetrachloride. After washing the precipitate free of carbon
tetrachloride with alcohol, it is dried at 110C. because the
mercuric sulfide is volatile at high temperatures (400+ tJ C.).
 GRAVIME'l'RIC METHODS 47
The reactions which occur may be represented in their SIm-
plest form by the following equation:
HgCb + H2S~HgS + 2HCl
271.52 34.08 232.67 2(36.47)
The same principles are employed for the estimation of mer-
curic oxide in ointment of mercuric oxide and for the determina-
tion of the mereury content of such compounds as ammoniated
TADLE rrr.-O~'li'ICIAL SUDs'rANeES ASSAYED BY PRECIl'ITATION AS
SULFIDE

Sumple,
Rcsi- Fac- Offieiall'cquiremcnt,
SUUSLILIll\(] Gm. or
due tor per ecnt
eo.
~---'-""--
----
U.S.P.
Bismuth lend potas-
sium tlLl'trate, .... 0.4 Bi 2S3 0.9063 Bi 20 a = 71 to 75
Morbltpholl ......... 0.5 JIgS 0.8622 Hg = 33 to 34.5
Mercuric ehloride,
poi~on tahlets of,
li1rgc, . , ...... , , . lOa IIgS 1.167 HgCh == 0.45 to O.55 b
Mercuric chloride,
POiflOll tlLblcts of,
small...... , ...... 20 a HgS 1.167 HgCb == 0.1125 to O.1375b
Mercuric chloride ... 0.5 HgS 1.167 HgCh = 99.5
Mereuric oxide, oint-
nwut of ........ , . 10 HgS 0.931 HgO = 0.9 to 1.1
Mercury, !tInll1oni-
a.ted .. , .......... 0.5 HgS 0.8622 Hg = 78 to 80
Mereury, amlIlolli-
[Lted, ointment of. 1.5 HgS 0.862 Hg = 7.1 to 8.7
N.F.
Bismuth, giycerii,B of 3.0 Bi2S~ 0.90G3 Bi,Oa = 12.5 to 13.5

a Numbe!' of tllbl"tH.
h GmmH pill' tliblet.

mercury and merbaphel1. after the mercury has been converted

into a readily ionizable sn.lt. An n.nalogous procedure is employed
for the estimation of bismuth compounds.
Questions and Problems
1. Show how the factor for the conversion of the weight of sulfide obtained
into an equivalent weight, of mercuric chloride is derived.
48 (JUANTITA'l'[VE 1'Jl.tillMACEU'l'ICAL CHEMIS'l'l~Y

2. How much mercuric Imlficlc wallhl 1 GIn. of mOl'baplwu contrtining :H

pel' cent of nwrr:nry yield '/
3. What ,,(llverd, is employed to remove "1.]10 ,~lllflll' jn the tl.H,sny of (lI)
lllmbrlphclI, (Ii) IJi~llIlllb [\,]\(1 potllHsinlil l.al'l,r:ILn, (c) allllllolli:d,celllle1'(:Ill'Y'i
4. lj~xplaill how t.ho pn:snJH.:e or hyd I'CldtlOl'ic :wicl Lmuhe j,o repreSil (,he eon-
c:enLraLillll of suHidn iOIl ill 1111: a;lol:LY 0[' IIIC'I"l'.lIl'ie: e'.h!orkh:.
5. What spceitll pn)(:edl1l'1: I::; cll'll'loyc([ ill thn dl,j,(>rmilllLt,ioll of IlIP1'C:Ul'Y
in (a) ointment, 0[' ulllIllollinl.nrl 1l1(!rClll'Y and oilll.lllellL or lllln'lmrie ()xie[e,
(Ii) hl'gc poison tnblot.H of JIl(;n:llrie c:hloride'/
6. :How would you prm:oe:d (,0 siLlJlpln (a) II t(!ll-pound jnr of !tl1llllolliatcd
lllerelll'Y oiulment" (II) II, hoLl.ln COlli (Ii nillg ] ,000 Hllmll i>oisOJl j,ul)lels of
mIJI'I~lli'it: chloride?
7. A 5 cr. H:UlIpl,) of gIYI'.(;J'il,o oj' bismnl.h yil'lclc)cl (l.!)004 UJlI, of .Ili~S:I.
(;[I!cllbtu IIll'. pel' e~c.;Ilj, of Bi});,lY! V ill Ilw I-';l,v,)ul'it.n,

Exercise 7
Determination of the Purity of Calcium Glycel'ophosphate.--
The ealcilll1l is prul:ipi t.atecl as ox:dnt,(), ignited, and woigll()d afl
oxidn.
Object.--Assay of Calcium Gly(:e]'opho:-;pha].(~.

Materials Requil'ed.-O.J, Gil\. uf CJ!I]".illlll g;]YI:t:l'Ol'hoNph:t(,e.

20 ce. of 5 pt:l' CCli t acetie :Ieiel.
AmmoniuJ\l oxalnte '1'.8. (3.5 Gill. !lmmoniulll oXIll:t.t.e ill 100 oe.).
Procedure.--l. " Dry about 0.1 Urn. or Cale:illlll c.:lyeur()ph()~!Jhf\te t.o
COllstanl. weigh I, HI; 1300., weig;h [\()cumlcly, dii'soh'" ill 20 nIl. of 1111 :U.1I1COUH
solution 01' noutio HeidI ill 20, [lml Held at) c~(',. of disl illc:tl \\'fllnr, II.c:nl; tile
mixture to boiling, and ndrl (HI DXC()SH of :unHHIlIilllH IlX:lja1.0 '1'.8."

Cl1leiuni. glyeerophoHplmte is hygl'oseopio apd must be: dried

at 1.:30 0 to remove absorbed '\vator. 'flte salt is soluble in abouli
50 pnrts of water at. 25, )Jut at higher tnmpemtUt'cR it be<:ome~;
less soluble. Acetic aeid render:, it m.ore solnhln, but tho (:aI(~illm
OXlLbte formed in tlill precipitation 1'Pil.(:I,ioll is insolnhl() in :weLie
acid solution. AnllflOnillln nxnlu.Le 80111 Lion is Helde!l in t'xeOSB
because it rcduces the sulubility or ealeinm oxalate through
common ion dTeet [\,nd tends to holLl rtny m~tgllcsinm pl'l~sclll) in
,;;olution as a double mmnoniulll llw,gncsiwll salli. }lreeillitati(Ju
should be carried out in lloilillg solution, u(lcling the prc(:ipit,aLing
agent slo'wly with eons(;an(; stirring to eauso the formation of
coarse crystals. ']'110 Pl'ceipit;[tiie is best washed IIY deoanttLtioll.
2. "Collect, the resulting precipitnt.e, wash, dry, nIlel ignil,1) to I~Onl'lt.allt
weigh!;, using f1 hl!1Ht; 01' high t.empemtllrc hUl'Ilor,
 GlUl VlMW1'lUC ME'l'HODS 49
"ElICh gl'nm of the residue (calcium oxide) is equiVl1lent LO 3.N73 Gm. of
CaCaHr,(OH)cPO.!."

After qnantitatively trallsferring the preeipitate to an ash less

filter antI W~t~:hillg, the filtrate sholll(1 be examined to determine
wlwtllcn' any erystals h:we passed through, siuee caIeiulIl oxalate
frequently comes down in illl{!-gl'ained, difficulLly filterable erys-
tahl. 11' crystals are observed in tho filtrate, the latter should be
repuRscd through t,hl) flltel'. The preeipitate mny be ignited by
the wet, method, [1S dns(:l'ibcd on page 2ll. The ignited precip-
j'Ln,l,() shol1ld be eooled in [l, desic:c:atol' nnd wl)ighed as SOOll as
possible, Rillne c:dc:illlll oxide absorbs moisture and e:\,rboll di.oxide
rapidly frolll the nil'. Upon strong ignitioll c:nJc:iulll oxabtn
forms ~:aleiulll oxicle as shown hy the following eqnatiolls:

C aH[)(OH)2CnP0 1 + 2CELCOOR-->Ca(CILCOO)~ +
210.15 C 3H[j(OHhH 2 PO'1
Ca(CIIaCOOh +- (NH1hC~01-->CaC20,j + 2CI:IaCOONH.j
Ca'cM),j->CnD -I- CO~ + CO
51i08

Employing the above reactions as a basis, ealculate the pc

eellt of CaO in the sample.

Questions and Problems

1. Is ealeiulll oxa!n.to more Hoillule ill hoi, thml in cold water?

2. Explaiu how an ex.ee~K of :lIlIlllouiulll oxalate (](,el'mwes the solubilit.y
of calcium oxnJrtt.e by eommOll iOIl dleUl;.
ll. Why shlluld pre<:ipitation hI) (:arried onL slowly from 110ijing solution.?
4. Explnill how to ignite :L pl'CH1ipitnLo hy the wet method.
6. Why ,:llOlIld the ignition l'esilillc be weig;1l(lcl as mpidly as po~;;ible ?
6. How lnueh enJeiulfI oxide i;hcmld 1m obtltined from 2.5 GIll. of purc
(laiciulll l'Yl'oplwsplut {'(J?
7. Briclly expbill how i.olletcl'lllinc til[) ci,tlciulll coutent. of C:,leiuUl lrtctatc
gl'l1vimet,l'ielllly.
8. Write the eqlwtions for the l'ClwtiollS that take place in each of the
following ofIicial nssaYH: (a) zinc :t,cetl1te, (I,) manganesc glycerophosphate,
(0) zinc phonolsulfonate. .
9. Show how the factor in each of lhe assays under question 8 is derived.
50 QUANTITATIVE PHARMACEUTICAIJ CIlElvIlSTUY

Exercise 8
Determination of the Aluminum Content and Purity of Alum.-
The aluminum is precipitated as hydroxide, ignited to oxide,
and weighed.
Object.--Assay of Alum.
Materials Required.-l GIll. of alum.
1 Gm. of amrnonium chloride.
Ammoni!t T.S.
Procedure.-"Dissolve about 1 Gm. of Alum, accuratc1y weighed, and
a,bout 1 Gm. of ammonium ehloride in 250 ceo of di:,;Lillcd wnter. Heat the
solution t.o hoiling, and add 11 Hlight eXeOi:lH of llnllllonia '1'.S. to pnleipiLnj,e
aluminum hydroxide. Collect tho preeipit[1te OIl It HUer, wash thoroughly
with hot distilled water, dry, ignite strongly, and weigh. The weight of
the aluminum oxido so obtained, multiplhld by 8.8114, iIldicntes its cquivnlent
in AINH4(SO.)2.12H,O and when multiplied by 9.307, indicates its equiva-
lent in AIK(SO.)d2H,O."
Ammonium chloride is added to the dissolved alum to prevent
the formation of the colloidal form of aluminum hydroxide and
to prevent the precipitation of other hydroxides of metals,
as magnesium, which may be present as impurity. Ammonia
water is added to precipitate the aluminum as hydroxide. The
ammonia water should be freshly distilled, since upon standing
in glass containers it dissolves silica, the presence of whieh leads
to high results, the silica being precipitated and retained in the
precipitate. The ammonia water should be [ldded in very slight
exeess, since aluminum hydroxide is slightly soluble in strong
solutions of ammonia. '1'he preeipitation is conducted in boiling
solution to convert any colloidal aluminum hydroxide into lnrge
particles and to secure a coarse-grained precipitate. The solution
should not be boiled after complete preeipitation, for ammonium
salts become aeid in reaction upon prolonged boiling dnc to loss
of ammonia, and the resulting acid solution would dissolve some
aluminum hydroxide. Aluminum hydroxide forms a slimy
precipitate difficult to wash. It is most easily washed by deean-
tation, using a hot wash liquid prepared by adding a drop of
ammonium hydroxide to hot distilled water. As mueh of the
supernatant liquid should be passed through the filter as possible
after the third washing before transferring the precipitate to the
filter, since the nature of the precipitate renders filtration very
 GRAVIMETRIC METHODS 5J

slow. The precipitate ignited by the wet method should be

heated strongly, preferably over a blast lamp, after the ash has
T,UH,ju IV.-Ol!'l"IG!AL SUD STANCES ASSAYED DY IGNITION 'ro OXIDE

Sl1lnple,
F(f.c~
Substl1nc<l Gm. or Ucsidue Offioial roq11irement, per cent
co. tor

._- - - - --- .
U.S.P.
AluIIl, n.ll'unonium .. , .... 1.0 AhO, 8.8\)4 AINH.(SO.),.12R,O ; 99.5
Ahull, amnlOlliuln, (lxBic~
c:>1,,,d ................ 0.5 AI,O, 4.G53 AINU.(SO,), ; 96.5
Alum, plltllflt'lillffi., . . . . . . 1.0 .11.1,0, 0.307 AlK(SO,)"lZH,Q ~ 99.5
Alum, POll18sium, cXHlc ..
cllt.cd ............... 0.5 .11.\,0, 5.060 AIK(SO.), = 96.5
Bismuth B\lb~.[llbonlLt,e ... 1.0 BizO, ...... BioO, = 90
Bismuth BubglLlI!1tc ...... 1.0 m,o, ...... BhO, = 52 to 57
Hi"muth subnit.rllto ..... 1.0 BizO, ..... , RhO, = 79
Bismuth Bub"nlicyillte . 1.0 13;'0, ...... m,o,
= 62 to 60
Cn.Icil,lnl CI'cosotu/je ..... 0.2 C"O , ..... CuO = 40 to 50
Zinc acetate . .... , ...... 1. () ZnO 2.261 (CH,COO),Zn = 83.16 to 87.32
Zinc clllarido ............ 1. () ZnO 1.071 ZnCh = as
Zino oxido, ointment of .. 2.0 ZnO .. "" ZnO = 19 to 21
Zhw BulilltO ............ 1.0 ZnO 1. [184 ZnSO. '" 55.86 to 58.63
N.F.
A1uminmn n.cet,nte, Bolu ..
tiOll 01 .......... , ... 0.0 AhO, 4.0031 AI(C.H,O,,. = 4.8 to 5.8W IV
Aluminum ohlorido ...... 0.5 .11.1,0, 4.7369 AICIa.OR,O = 95
Aluminum ehloridc, 80\11-
tion of. .............. 5.0 .11.1,0, 4.737 AICb.OH,O - 22.5 to 27.5W/V
Aluminum Buhacotlttc,
solution of. ........... 5.0 .11.100, 3.1788 Al(C,H,O,J,OIl = 7.5 to 8.5 W IV
AluminulIl sulf'lto ....... 0.5 .11.1,0, 6.r,37 AI,(SO.),.18H,Q = 09.5
Bismuth mugn,\ ...... 10.0 Hi,O, """ Bi,O, '" 5.6 to 6.2W/V
Bismuth Bubcnl'bonltte,
tnblllts of ....... " 3 .0 Bi,O, ...... nbO, '" 83 to !)7 b
Bismuth Bullgall,lte, tab-
lets of ................ 3.0- BbO, ... ..
~ BhO, = 48 to (ii'
BiBm11th Bubnitl'ttte, tab-
lets of ............. 3,0 BbO, ...... Ili,O, = 73 to 85 b
Hi"m uth BubBlllicylttto,
an1Puls of ........ " .. 1.0 Bi,O, ...... Ili,O. = 57.6 to 70.4 b
CalciumglyccrophoBpitl1te 0.4 C"O 3.7473 CnC,H,(OH),PO, '" 98
MltngllIlcse citmte, Boluble 0.5 Mo,O. 2.3728 [C,fhOH:(COO),],Mn, ,., 4.8 to 52
M"np;lJ,ncse glyccrop!toB-
plulte .......... , .... 0.1i Mn,O, 2.95 MnC,H,(OH),PO. '" 98
Zinc llhell(lIBulfonate .. 2.0 ZnD 6.8284 Zn(C,U,OSO.),.8H,O = 99.5

Weight of ingl'odiont Bought.

b Per cent of Inbeled amount.

become white to convert any traces of basic aluminum sulfat.e

to the oxid.):
52 QUA.N7'I'l'A:I'IVE PI1AIUJIACIWTICAL ClllE!JllS'l'RY

2AINH.l(SO.lh.lzn..n + (iNH.OIT-):~Al(OH)a + 1(NIT'I)"SO.\ + 2.{n~,O

2 X nOD
2Al(OH),--->ALO" + :\11,,0.
lUl.04
\)()I\,[Jl:101.\ll,: :;1::1
10U14,t 0' \loti.net
x = 8.8fJ.:L em. alum erillival('.nt to 1 Gill. Al~O".

C~dt:Ulnto the plll'it.y of the ahlln and tho Vfll' nent of n.lllminmn
in the sample.
Questions and Pl'()bl~!ms

1. Why iH IIJmnOJlimll l'hlol'ido add!'!l 10 Iho all1l11 HOluLioll proVi01U-l 10

jln'eipi h\'t,iOll'l
2. Why ,;1lOuld a l:trge (1:Xl:eH~ or 1<H1111011i1lIII hydroxide he lLvnirlnrl?
3. Indicai.) hy reac.lj()IlH how amlllonium HaHH in 80lnl ion may beoolllo
neicl in l'etwtioll 11}lOJ\ pl'olongt!d boiling.
4. If an unlulOwll Hample of IllulU yic!lch; 0.125 GIll. of AL{)" llpon assny, 1.0
how mueh AINTLt(SO'I),.12H,O, AINII.t(f~O.,)" Hllrl AlJSO.I),.12H"O,
l'OSpcctivnly, i;,; I ho AlI)" oCl11ivalent?
6. A 8.0000 Um. Hlllllpie pl'np:l.l'l'd hy ]lowilprilll,!; 20 lahletN of hi.~IlIllth
RUbllitmi~c which weighrfl G.8420 Gm. yielded 2.8ti87 Chil. (Jf Bi,()a. CnIC'lI-
bic lhe 1\,J'I\OImt of Hi tend or Hi 20 a (~(m(l\ill('.d ill (~tl.Ch (.[lh1e!,. U:t]en!tl(tl (111\
pel' cenL of Hi aud of Bi,O,1 COIltlliJlm[ in en('ll tnblet.
6. Write eqll:Ltiolls for the rnactiollil tlmL I.nkl1 p11\(11) ill tho a~~:ty of oilll,
mont of zinc: oxide.
'1. How 111l1f:h P\U'l) 7.ill(l Rulfntn would hl! rnqui['('cllo fO],Hl O.8n20 (lIn. of
7:110'1
8. A snmple of Holnl)le lllfl,lIgn.lleRC c!itraic; weighillg 0.502,[ Gm. yidtlecl
0.2744 Gm. of Mn"O'I. WIlli" pOl' CllUlt of Hl:lII[~nl[()Rl1 dicIt hn :<tIlnple COli (.:tin 'I

Exercise 9
Determination of the Magnesium ill a Magnesium Salt.-The
lllagnesium is pl'ueipiLntecl as mrtglH';,iullI amlllonilllll phosphate,
washed, dried, ignited t.o pyrnphosphnJf\, and weighnd.
Object.-AsRay of Solutioll of' Mngnpsililn Citmf.e fot' Mag~
nesiull1 Oxide.
Materials Reqllired.-l0 erl. of soln1iolt of mngllfl:;illlll (:11,1':11.8.
2 eG. of hydroehlorie aeid.
20 ce. of sodium pho:;plwtc '1'.8. (12 (Jm. Na,,,IIPO~ in 100 ee. of diHWh~d
w!1tcr).
Stronger ammonia '1'.8. (/l.t lcm;l~ 27 )lCI' cent NH:I) ...
Ammoni:1 '1'.8. (9 to 10 per cont NH~).
Procedul'e.--l, "Transf01' to :1 benker of about 200-ee. cnpnci1,y eXfletly
10 ce. of Solution of Magnesium Citl'ate which hUH btlen pl'cviollf:ily freed
 GIlLl VI ME1'RlC' METHODS 53
from excesflive em'boll dioxide by repcated pouring, Add 100 ce, of dis-
tilled wll,tor, 2 ce, of hYllror;hlorie Hcid, 20 ee, of sodium phosphate '1',8"
,tlld 2 drops of Illct.hylred '1',S, Add tlInmonin ']',S, n few dl'0ps at a time
with eOllBtant Al.irrill~ until the solution becomes faintly ycll(Jw, Allow the
mixture, to stand for ten Inillut;eH, fldd 40 ce, of stronger mnmonin, '1',8, with
{)OIlstant stil'rillg, a]\[l BIlow Lhe mixLlIl'o 1.0 st.and for two hours 01' over night,1I

The sample is lllt)t\,sul'ed aftor removing most of the earboll

dioxide gas by pominfs to prevent loss clue to effOrVeflC(~nCe,
Whon sodium ph()sphatl~ is dissolved in wator, the phm;phorie
aeid ionizDH in throe slin,ges which mny be represented by the
following oq LLl1tiollS:
HaPO li.-~H+ + lI PO:j
2
H2PO;i'=->If+ + lIPO:!,
npo1':'~---)H+ + PO~
The degree of ioni:t,ntinn bneomes less for ()[wh sllcccssive stage,
The relative f\,Ulollnt of Jl07, HPO;;, H 2pey;-, and HilPO,1 in
equilibrium wiLl! each oLlwl' depends upon the hydrogen ion
eoneontl'atioll of LllO ~ulnti()ll, In [L basie Koilltion, the PO~ and
IIP01' ions pl'cHlomiuaLe, l%eh of the ahove species of phosphate
ion whieh exist in the solution of phosplmte reacts with Mg++
ions to form the corresponding mfLgnesiulll salts, all of which
arc morc 01' less inRoluble in wu.tel', TIlllA there may he formed
Mg:l(P0 1 )z, MgITPO,1, alld IvIg(H 2PO.1h The addition of a
solution of NIl,jOH l'npl'OSSCS the coueentrntion of H 2P04' iOllS
ll,nd provcnts tho furm:ttion of M.g(H 2PO,lh In the pl'eserwc
of the u.rmnolliutn ion, the salt, is procipitated as 1\oigNH1P0 4,-
6H 20, If the Rolul;ion is tuo basic, IVIg(OH)2 may be precipi-
tated, The precipitation of both (Mg 3P0 4)a and JVIg(OHh is
prevented by tho pl'OSCIWe of l'Lmnl,oninm snIts which, through
common ion effoot, aot as [t buffer ltllcl prcvent the OlI ion
eoneontl'n.tion from becoming too gret'Ll;. The hexn.hydra,te,
MgNH 1P0 4 .6H 20, forllls in I'elntively coarse crystals on standing
for throe or more hours at room tmnpon'Ltmos, At temperatures
above GOC, the mOllohydmto, MgNH.1POdI 20, is formed.
2, "Collcc'C tho precipit.ate in a weighed Gooch orucible a.nd wash with a
mixture of 1 volume of ammonia T,S, and 3 volumes of didtilled water until
free from chloride, Dry and ignite to !l. eonst.!l.nt weight. 'l'he weight of
magnesium pyrophosphate (Mg 2P 207) so obt!l.incd when multiplied by 3,621
gives the amollnt of magnesium oxide represented by 100 cc, of t.he solution,"
54 QUAN'l'I'l'A'1'IVE PIIABMACEUTICAL CHEMISTRY

The precipitate is washed with aqueous ammonia solution

until free of chlorides. In testing the filtrate for chlorides,
acidulate with nitrie acid before adding the silver nitrate solu-
tion. Upon ignition, the magnesium ammonium phosphate IS
converted into the pyrophosphate as follows:
2MgNH 1PO,j.GH 20---l-J\I[g2 P 207 + 2NHa + 13H 2()

For the purpose of making ealculations the equation may be

represented as
2MgO->Mg 2P 20{
2(40.32) 222.68
Thus it is evident that each gram of Mg ZP 207 is equivalent to
80.64/222.68 = 0.3621 Gm. of MgO, aml, since 10 ec. of the
TABLE V.-O],FICIAJ, SUJlS'l'ANc:r~s ASSAyrm GUAVIME'1'HIGAJ,LY AS
PYHOPlIOSPHA'rm

Sample,
OHicin.ll'cquil'ClllenlH,
Substance Gm. or Residue Factor
per cent
ce.
--- -- -
U.S.P.
Magnesium cit-
rate, solution
of ...... , ..... 10.0 Mg,P,Ol 0.3G21 MgO = 1.6 to LOW IV
Magncsium sul-
fate .... , ..... 1.0 Mg ,P ,0/ 1.081 MgSO, = !l\l.5
Sodium phos-
phate ......... 0.3 Mg,P,Ol 1.275 NII 2HPO, = \18
Sodium phos-
phate, exsic-
cated ......... 0.2 Mg 2P 20 7 1.275 Na,IIPO, = n8
N.F.
Magnosium sul-
fate, ampuls of 0.2" Mg,P,O., 2.2138 MgSOmI~O = !l5 to 105b
Sodium phos-
phate, solution
of ............ 5.0 Mg 2P 2Or 1.275 NI12HPO. = 30 to 4:1 If IV

" Weight of in(lrerli~nt sought.

b Per cant of J!,belou amount.

solution was used as sample, 10 X 0.3621 = 3.621 Gm. of MgO

in each 100 ce. of solution.
 GllAVIME1'llIC METHODS 55
The pl'ineiples involved in this exercise may be applied to the
analysis for magnesium in all soluble magnesiuIU salts. Con-
versely, phosphates may be determined by essentially the same
method, magnesia mixture being added to the solution containing
phosphate ion. The arsenate ion, AsO:, lllay be precipitated in
like manner to yield MgNH 4 AsO,j. Salts similar to MgNH 4PO,1
TAm,E Vr.-SOME O'l'HEH OF'FICIAI, GRAVIME~l'lUC ASSAYS

8[1,1111>10,
SubHtt~n{l(l Gnl. or Residue OJIiainl requirement, per cent
cc.
---~-~-- ----
.. --.---~-

U,S,P.
Collodion .. , ........ , . 10.0 Pyroxylin Pyroxylin = 5.1 W IV
ClLrnpltOl', linhnCI1t of .. 5.0 Cottonseed oil Camphor = 19 to 21
Cu,ruphor, spirit of ... .. 25.0 CI,mphor dinitro- Camphor = 9.5 to 10.SWIV
phenylhydrnzidc
BI'ythrityl teirnniil'nto, (),25 Erythrityl j,etn.- C.H,(NO,). = 17 to 53
diluted. nitmtc
Gold chloride, R, ...... 0.2 Au Au = 47
Io(linc, cOlnpound, solu-
tion of (for K1) ...... Ii. [) KI RI = n.5 to lO,5W IV
Iodine, tincLul'e of (for
KI).'....... _ ....... 5.0 In In = 4.5 to 5.5lV /V
Iodine, tincture: (Jf, mild
(for NILI) ........ , . 5.0 NILI NaT = 2.1 to 2.5W/V
Molyhdic IUlhydddo, R 0.5 l'bMoO. MoO, ~ 99.5
I'L\l1lLdou~ chloride, B. .. 0.2 I'd I'd = 59
PILttinio "hlori<l", n .... 0.5 Pt Pt = 37
N.F.
CIl"ruphor I lunplIIH of ... 5.0 Ampul oil Camphor = 93 to 103.
Phenolphti1l11(liu, tl1blc~tB
of. ............. , .. O.G[jA Phenolphthalein Phenolphthalein = 92,5 to 107.5.

I Amount equivalent to.

Pllr "ant of tho ItLbeled I1mount.
R. = l'{HLgent.

are formed by zinc, cadmiulll, and manganese. Under care-

fully controlled conditions, the same general principles may be
made the basis for the estimation of cadmium, manganese,
phosphorus, zinc, and arsenic.
QUestions and Problems
1. Explain each sLep in the procedure for the assay of sodium phosphate
and writ.e equatioIls for all reael;ions that take place in the assay.
2. What is magnesia mixture T.B. ?
3. Under what conditions is Mg++ precipitated as MgNH.P04.2H 20?
[iii QUilN'l'FflM'1VliJ PllilJUUAC1EU'l'1C'11L CllliJMISTIlY

4. In the a~s!l.y of ltmpulH of lllagncmium sulfate labeled I< eac:h 2 ec. C:Oll-
t11ins 1 Gm. of lllal~n()sillIn r>Hlfnte," how would you Hmllplo It shipmellt eOll"
tnining 1 g;J'OHl! of iLlll)lllh:? How JIllleh of l,jw :LlllP1l1 RO]IlLioll Willi]!] yon
mOIItlU!'(l fur mwh aRsny smnplu'/ What miniltlUIllIIIlC] lllaxiJtllllllllTlJour\tS of
magrulHiulll sulfate t1lig;hL Iho mllJlIlI~: (\ollkdn in Dlle]t ouhie IIOllLillloter !"Illd
meet 1.110 oHieinl j,()1UnLIl(\O I'c:qllirc:lllclll t.H 'I
Ii, A ,w,Inplu of Ho,lilllll plIOHph:LI,(J w<:igliillg O.:l!i(i(i Um., when 11ricIl to
l!l)llsLnnt weilsht ai, nOne., wpiglll:(1 n. t ()'12 CllL allli UpOll m.iHiI.y yialtlod
0.11\)1 Clil. of .Mg~P~OI' Calnuiato the IH.'I' C\1\1I1. of 81H.liuIll pllllHplmto IIlld
of pllOHphol'llR itl Uw origillal Nlllliple ail'] ill (.}l(\ ltlOiNtlll'(:-fl'eo ~llllljJlll.
VVlmt pOl' eeJlt of ltloi<ll,ul'o did Lhe orig;iIml Hample contain'/
Other Gravimetdc Assays,--NuTlw],OllS gravimetric: methods
of assay oiJlwr than tlios() c01lr3idered ill tbis eitapti)r are used in
the determination of offieinl suhstances. Thosn whieh require
a i-lpeeial tcdlllique, snell as the gravimetric assay of alkaloids,
ILre treM,(][l in rmlmeq1Hmt ehnpters. Som.e of tho ()fIicial gmvi-
metric assayR, snch aM Che OlWS for alkali iodides, whieh involve
onl y evapora.tion, drying to eOllst:1llt wei gltt, , and weighing, are
ill( iea,ted in the table on page 55,
 CHAPTER IV
PRINCIPLES OF VOLUMETRIC (TITRIMETRIC)
ANAl,YSIS
In volmnetl'ie (tilirimetl'ie) analysis, or analysis by measure,
the quantity of an elcrnent, compouent of a compound, or com-
pound in [l, wcigJlcd "lnmple is nutcle to react with a measured
volume of a rcagent solution of known cOlwcntl'ation. Since the
volumc fLnd cOlw()llLratioll of the l'cfLgenb Holution required to
eomplete n, defillite reaeti()ll nre kllown, the amount of fmbstanee
entering into l"(1ll.etion wiLli the reagent solution can be calculated.
Thus the dllorido ion enntont in a soluble chloride i8 determined
by c1iRsolving the chloride in water B.nd [1dding silver nitrate
solution of known emwcntmLion until practically all of the chlor-
ide ion IU1H liGenl prec:ipit;nLnd as silver chloride. From the volume
of silver nitrate solntion onnsumed, the weight of chlorine in the
sample and. t.he purity of the Rolnble chloride can be calculated.
Definitions.--Tho Ol)()l'atioll whereby the eoncent.ration or
vltlu() or n, BoluLion for n specific l'Clwtioll is determined is tcrmed
st!l1ut(t1'IUzation. For cxmnple, a known weight of pure sodium
tarbonate dissolved in water, ~when made to react quantitatively
with n lUcaslU'od volmne of hydrochloric t1Cid, gives the data
rccIltircd to calculate the eonccmLraLion of hydrochloric add in
the solution. The hydrochloric acid solution thus stLtllclardized
is ealled it standard Hol'iltion, i.e., a 8olution wllii,d~ contains a
known 'We/alit of j'cIC(jcnt 11G1' 'unit of VOllt'llW. The pl'()cess of add-
ing a sta.ndard. solution or reag;ent in measured qnantity to a
flubstn.nee until the end point, of rmwtion is observed is called
titration. COllscqllently, volurnotric methods are often synony-
mously tonned titl"i'llwtric 'lI1ctlwds. rritmt.ion rnn.y be conducted
drectlll, 1:.e., n. stmldawl aeid is addecl to [1n allmli ltntil tho end
point is re[l,ehed, or 1'esiclually, i.e., [111 excess of standard a.cid is
added to the unknown ttllmli and the amount. of excess acid is
determined by titl;at.ion wit.h standard allmli solution. The end
point is the 11l"actieal poin1; at whieh t.itration is stopped when
57
58 QUANTI'l'A.'l'IVE PHARMACllJU'l'ICAL CIIRMIS'l'RY

the reaction is eomplete as shown by a pl1l'tieulm indicator or

other device llsnd for that purpose. The end l)oint Hhould not
be confused with the stoichiometric TJOint, i.e., the point whieh
marks the addition of the calculated equivalent of rel1cting sub-
stance demanded by the equation.
Volumetric Apparatus.-The 11pparatus required for volumetric
analysis includes the analytical baJa,nce desol'ibed under Challtcr
I and a number of accurately calibrated instruments as well
as beakers, flasks, funnels, etc. Volumetric appttl'atus is of two
types: that made to deliver a defmite volnme of liquid, as burettes
I1nd pipettes; and that made to contain a definite volume of
liquid, I1S volumetric Hasks and graduated cylinders. The tem-
perature specified in the U.S.P. for volumetric measurements
is 25e., while the Burel1u of Standn,rds has adopted 20 o e. as the
standard temperature for glass volumetric appl1l'atus. It mat-
ters little at what temperature the instruments a,re calibrated
IlI'ovided the Hamc temperature is used for all, and 0,]1 measure-
ments are made at the temperature for which the instruments are
calibrn,ted.
Units of Capacity.-The U.SP. liter is the volume occupied by
996.04 Gm. of distilled water at 250., weighed in air with brass
weights, with the barometric pressure at 760 mm. The official
cubic centimeter (ce.) represents the one-thousandth part, of the
normal liter. The normallitel' is defined as the volume oceupied
by a kilogram of water weighed in vacuum at 4e. The one-
thousandth part of a normal liter, termed a rnillilitel', and the
cubic centimeter, are both used as the unit of capacity. The cubie
centimeter, abbreviated as "cc.," which has been adopted aB the
official unit of capacity, is not exactly the one-thous::mdth'part of
a normal liter, since one milliliter = 1.000027 cubic eentimctel's.
The difference is so slight, however, that it beeomes negligible in
all volumetric measurements.
Burettes.-Graduated glass tubes of uniform bore throughout
the whole length used in the measurement of.variable quuntities
of liquid are termed burettes (Figs. 9 and 10). They are gradu-
ated i~to cubic centimeters and closed at thc bottom by a glass
stopcock or a short soft india rubber tube provided with a glass
tip and pineheoek or other suitable deviee to eontr01 the ouWow
of liquid. The official requirements for burettes follow:
 PRINCIPLES OF VOLUMB'l'RIC ANALYBIS 59

"All burettes should be provided with glass stop-cocks and

calibrated for usc at the standard or other temperature at which
they arc to be used. Burettes provided with rubber tubes,
pinchcoeks and glass delivery tubes in plaee of glass stop-cocks

PIli. U.-But'ottc with g\aFJS Pm. lO.-Bul'cttc with

st,O[loock. pinchconk. (Mohr bu-
rette.)

may be llsed for potassium and sodium hydroxide solutions and

such other solutions as are best used in this type of burette. The
p;lass tips should be from 2 to 3 em. in length, gradually tapered
and slightly bent. The rate of outflow should be regulated so
that not less than two seconds should be consumed for each ce.
delivered. When completing a titration, the tip of the delivery
tube should be touched to the wet inner surface of the receiving
vessel and the solution again stirred."
Burettes provided with glass stopcocks should be tested for
leakage. A lubricant such as pej;l'olatum serves to prevent leak-
60 (JUAN'l'ITA'1'IVE PIIABjIIA.CEU'l'IGAl~ GII1?M18'my

age [mel permits casy opcmtioll, but an exeeRS of lllbri(mnt :,l!ould

never be used, Those provide!l with [l rubber outlet (Mohr
burettes) should neven' be used for oLlier thrw nlknli solutions,
The outlet Lip of eiLilCl' type of burette Hhuulcl be of HIWh lliameter
and tapor as to [lcmuiL tho ddivc~I'Y of:}, drop tho volume or which
is consiclern.l)ly l(\SE: titan t.hat whieh e~m be [wc:olUlllodated
bet.ween LllfJ Jincst grarluaLions of the Hoale wit.h
~~~
'-'~ wliie JI tllO blU'ntLc ii:J InaI' IWIL
Reading Burettes.---The snrfn.r:e or liqnidA in
nan'ow tubes is n,l WtLyS cl1l'ved due to <:npillarity;
\1'{, the Rllri':L(~e or 'It/,cn.ISCI18 is always eonenve when
the JiqlLid \vets thu tube and convex wlunl it; does
not. l-icLMlilllJ,s should ttlwttys he nuttle at tho lowes\,
11l)int; (If the meniscus, that is, tho graduution nltLrk
whieh eoineid()s with the bottom of the curvo, excopt
when .measming highly culored liquids. The eye
lUUS!; be on the same level as the lllenis(;us, other-
wise error will be inLl'odnoed dun to pa.rallax (see :Fig.
FIn. ll.,- 11). Numerous dovieeFi have been employed to
gffunt-, of
parallax in aSRist in reading bnrettos eorrcd.ly. Onn S\lell device
l'cndinp; 111l- l'eeOlnmended by the United StaLus Bureau of
l'cl;tflS.
Standards is It eolbl'-dtapod sectiolL 01' blank rubl)er
tubing, cut open at one side and of l\, size whic:h fits the blll'otto
tube so that it; can be moved. This hbek l'llblwl' t\lbing;, when
pl::wed immedird,ely below the meniseu8, rondors the Hl<miH{',\lS
profile dark and elonxly ()bA(~l'Vn.ble agaillAli [\, light lxu:kgl'OUll(l.
Pipettes.-Tho following reqniromf)IJts [01' pipettes have beon
adopted in tho U.S.P.:
"Pipettes should be graduatnd to deliv~r fit standal'd torn-
peraturu tho volllme inclioaj;ed. The suetion stem, Ahnutd 1m at
13ast IG om. long, and tho delivery tube not less than :i em. and
not more thall 25 em. long. The inside llin,met,()l' :d; Lhe Uftpacity
mark must not be less than 2 mm. It 11lUAt noli exeped t1 mIll,
for pipettes up to and including 25-ec. eapaeity, fj Dun. for 50-oe.
capneity tl,nd (j mm. fo1' 100-ee. and 200-ee. eapaeities. The
capacity mark: must; Hot be more than 6 em. from the lnJlb. Tho
outlet of any transfer pipette must be of such sizo th:11; the froe
outfiow for water shall last not more than one minute. Not lesH
than fifteen seconds shall be required to empty a 5-co. pipette,
 PRINCIPLES OF' VOLUME'l'RIC rlNALYSIS 61

twenty seconds for a 10-cc. pipette, thirty seconds for a 50-cc.

pipette, forty seconds for a 100-cc. pipette, and a minimum of
fifty seconds for a 200-ce. pipette. After filling, the liquid adher-
ing to the outside should be wiped from the stem. When empty-
ing the contents, the pipette should be held in a vertical position
l\,nd the outflow should be uIll'c\strictecl until the surface of the
water reachcs the upper end of the delivcry tube; the tip should
then bo touched to the wet surface of the receiving vessel and kept
in contaet with it until the emptying is complete. Pipettes
should never be dmined by blowing into them unless, as in the
caso of the OstwfLlcl pipcttcR, they axe cspcci:tlly gmcluated for
\lRe in thiR way.
"The linlitR of ()t"1'Ol' pet"mitted ill the calibration of pipettes
shall he those aeoepted by the United States Burean of Standards,
whieh are [LS followR:
CC)IltClltK (If pipettes 2 ec. !j CC. 10 ell. 25 ceo 50oe. 100 co.
Limii. of cr1"01" ill er:. 0.000 C)' 01 0.02 0.08 0.05 0.08
Limit of C1"I"01" in Pf)l' eell[; 0.30 0.20 0.20 0.12 0.01 0.08

"It i." nssolltinl that in using' pipettes the samo procedure be

employed lL~ WliS used in the calibration."
In using Ll'fLllSfer 01' delivery pipettes (Fig. 12), they are filled
from the jut to a,bout 1. em. above the mark, then allowed to run
down just to tho marie The liquid jf) drawn up by sucking the
upper end of the pipette with the mClllth, but this procedure
should be !woicled and some other method of measurement
employed when volatile, corrosive, or poisonous liquids arc
measured.
Measuring Flasks.--The U.S.P. requirements for measuring
or volume(;rie flasks (Fig. 1:3) are as follows:
"Standard measuring Ibsks nl'C calibrated to contain, v"hen
filled to tho mark, 1000, 500, 250,200,100 or 50 cc. at 25C. '1'ho
necks must measure not IpsR than H mm. and not more than
20 mm. in diameter for lOOO-ce. capacity; frolll 12 mm. to 18 mm.
for 500-eapacity; from 10 rnm. to 15 mm. for 250-cc. capacity;
from 8 mm. to 12 mm. for 100-cc. capacity, and from 6 mm. to
10 mm. for 50-ce. capacity. The capacity mark on any of these
Hasks should be not less than 6 cm. distnnt from the mouth and
not less than 2 cm. from the base of the neck. The limits of
62 QUAN'l'ITA'l'IVE PIlARMJ1(!FJU'l'J(!AL OlIEMIS'l'RY

error permitted in tho ealibratioll of flasks shall be Lhosn aeeepted

by the United States Bureau of Sli::1lldarrls, which are as follows:"
Contents of flask 50 ce. 100 (:C. 250 ce. 500 ce. lOOO ceo
Limitoferrorincc. 0.05 O.OR 0.12 0.15 0.:30
Limit of error ill per cellt O. 10 O. os 0.05 0.03 0.03
Volumetrie flasks arc Hsed to make up RLnndard solutions to iL
given volnmc. Note that the U.S.P. requires that flasks be
graduated to contain and not to deli vel' the calibrated
volume.
Graduated Cylinders.-Gmduated cylinclers are
used in making approximate measures of volmno.
They m'e graduated to contain a given volume of
Equid at standal'll temperature. The insicle dimn-
del' of tho cylinder should not exceed one-fifth of the
graduatcd length.
Cleaning Volumetric Apparatus.-N ew volumet-
ric nppmatus should nlways be <:leaned before using,
and that in use should be eluttrwtl ns often H,H ncecs-
sary. The adherence of droplets to the wall of a
burette or pipette is positive evidence that the appa-
ratus is dirty. In a dean instrumont, the liquid
a drains clown uniformly wetting the
walls so that no droplets are obsm'v-
able. A warm cle::'l.l1ing solution (130-
clium dichromate in sulfuric acid),
solution of sodium hydroxide, or solu-
tion of soap powder is the best cleans-
ing agent to usc. Hot l:Iolutions
should be avoided when donning aecu-
ratcly ealibratecl apparatus bceause of
the possible production of [L permanent
change in volume caused by the hoat
FlU. 12.- and known as "thermal after-effect."
(a) Tmnsfor
or dolivery Calibration of Volumetric Apparatus.
pipette. (b) The' ordinary volumetric apparatus Ii' I ll. l:}.-
Mell.suxing Volumetri()
pipette. marketed is calibrated at various tem- fl!lsk.
peratures and is often unreliable, The
analyst should always check the calibration of volumetric appa..
ratus and make corrections if necessary.
 PRINCIPLES OF' VOLUME'l'RIC ANALYSIS 63

Burettes.-The calibraiiion of burettes llmy be performed in

various ways by means of special calibration devices such as the
Ostwald calibrating pipette and the Kiehl burette calibrator. In
all practiol11 work, however, the following method is satisfactory:
'rho dean burette previously tested for leakage is filled to the
zoro mark wit,h distillod water. The burette and water used
should be in the room a sufficient length of time before use to
[Lequire room temperature. A definite volume, e.g., 10 cc., is
allowed to run int.o n tared beaker. The weight of water with-
drn.wn is dnterminod [Lnd the burette is filled again to the zero
Inl1rk and then 20 ce. is withdrawn in exaetly the same way.
This operation is repeated for each 10 ec. until the 50 cc. mark is
reached, determining the weight of water withdrawn each time.
The results with corrections should be recorded in tabular form
;;howillg the rclation bctween the apparent volumc and actual
weight delivered to cnell rnnrk on the burette corresponding to an
interval of 10 ce. Weighings need not be more accumte than to
the seconu uueilllnl place, since that is the greatest degree of
accuracy with which the buret.te can he rcad. In practical
calibration, 1 co. mny be considered equal iio 1 Gm.
A buretto c!1libmtcd at 20C. should deliver 9.9718 Gm. of
water or at 2.5C. it should deliver 9.9604 Gm. for each 10 cc.,
since 1 liter of distilled wnt.cl' at 20C. weighs 997.177 Gm. and at
25C., 9HG.(}<1 Gm. when weighed with brass weights in air of 50
per centi humidity 11t stancln,rd atmospheric pressure. The true
volume for Olwh 10 ec. segment of the burette may be calculated
from the weights obtained and recorded on a convenient chart.
Pipettes.-Clettn the pipette thoroughly with soap solution
and denning rnixiillrO and rinse well with distilled water. Fill it
with distilled wat,nr at 20 or 25C. according to the temperature
selected :for ealibration, and allow it to drain into a tared iia,sk.
Weigh ~1Geurnt()ly and record the correct volume of the
pipette.
Flasks. -Clenn, dry, and tare the flask, placing the number of
grams weight corresponding to its volume on the opposite balance
pan and introduce water at 20 or 250. into the flask in sufficient
quantity to counter'balance the added weights. Mark the lowest
point of the meniscus if it does not agree with the original
calibration.
ti4 QUAN2'l'l'il'1'IVB l'IIARMACEU'l'[(JAL CII[I]MI8'1'j-tY

Sources of Error in the Use of Volumetric Apparatus.-Every

precaution should he takcn to avoid the following common
sources of error in the llse of volumetric appamtus:
1. Hin:'ie water adhering to walls of apparatus. The appn,ratuH
may he dried, but it is more convenient and just ItS uecnmtn to
wash it out with sillall successive portions of thfl liquid to he
Ilsed, discardillg ihn washings.
2. Oren-flu films ltnd dirty applwatus e:lllse il'l'ogulfLrity in tlte
ddivery of liquids and distor[, the meniscus.
8. Dmillage 01' nl'Lerflnw or the liquid adhering to tho vessel
walls to tho liqni(l helow l1eeUHsitaj,es that the time of drainage
be tho saltle as that use(l in the eoJibmt,ion of the illstrument.
4. Parallax HI1!St be avoided to secure propel' readings of tho
lewel uf tlle mClliscllS.
5. Variations in tempel'f1i;nro lead to elmnges in volume of
vessels and liquids. All llWLLSUl'OlfWlltH, therefore, should be
Iimde at [I, tempemtul'e dosely approximl1ting that n,t which the
apparatuR W!L8 calihmt,pcl.
G. Air bubbles tmpped benen,th the liquid snrfneo, especially
below the stopcoek in burettes, disphwe liquid.
7: Heat, such as supplied by hot solutions, eauses calibrn,ted vol-
mnetric apparatus to sufier a slight; permanent ehange of volume.
8. Most FiaHs when dissolved produce a ehanp;e in tcmpemture
with concomitant elw.nge in volume of the solution. Suell solu:"
tions should nevor be measured until they have nequired the
tempcrature at which the inf:d;rnments used to mcasure them wore
calibrated.
fl. Failure to use appamtus ill a manner npPl'oaehing as nearly
as possible that followed in ealibration.
Questions and Problems
1. DelillO Lite followillg te1'111S: (a) ~Llllldardizl1t,ion, (0) si;!tnda1'd sohil,ioIl,
Cc) end poillt, Cd) residual titration.
2. What is the oHicialliLer?
3. Blll'otteH provided wilh rubber tubes and pincheocks in place of gltLHR
stopcocks lnay be used to measure out sodium or potltSSiUIIl hydroxide
solutioll, but they should never be llHCcl in the lllC[t~urcmcnt, of soiutiollS of
lteids or oxidizing ltgents. Why?
4. A 100 ce. volumetric flask calibrated at 25C. is used to meltsure wltter
at 20"0. Whltt eorrect.i.on of the volume must btl macle? (See U.S.P. XI
table, page 623.
 PIUNC1PLBS OF VOLUME''1 ' RIC ANALYSIS 65
Ii, A volurnetl'ie flask c[llibmted to contnin 100 (~e. lIt 25C. and 760 mm.
bI1rom(lf,ri(~
prCScl1lrr, is lls(HI lit 180. find 7411 1I1Ill. baronwtrie pressme.
Cflleubte the volume of \Vn,j,el' eOllhlinecl ill Lhe na~k at the tempel'ature
and preH~\lJ'c ilt whj(~h if, is \lser!.

NEUTRALIZATION METHODS-ACIDIMETRY AND ALKALIMETRY

Aci(Uuwtry is L!l\') measurement of the qmtntity of acid in a

given sample by titration with It suitable alkali. 11ikali/ll,ctl'lJ
it; the mnaSlll'f:Il1[)llt of the quantity of alkali ill a givell sample by
titri1tioll with n f-luit:1hle aei(l. In eadl eaRn thn p1'i1l(:i pIe in vol v(~d
iR the HfLnJe, lU1ll)cdy, to add to the a(~id 01' alkali being detol'rnilWd
an (~q\livt1l(mt amOllnt of stnnrlnrrl f-lolutioll of alkali 01' aeid,
mspcd;ivdy. II\Olll the t1l1lOtIld, of ::-:!'alHlanl Nuhl1,ioll arMed, the
11IlHHmt of nlkali or :wi(l ol'igin:dly pn~i-Wllt is c:aiculttL('rl hy meaus
of the fitoielJiOlllpLri(~ nqlllttioll, The l)oln(; n.twhieh all equivalent
of rcmeting HulmtnlH:n haH been l1,(ld(~d to allother i-ml):,;1,ance if,
tennetl tltn doichimndrir point, c!JlI:ilmlener: l){)illt, or thcO/'elicaJ
end [Joint. It 'i8 thlLt l)oint Itt whi(:h Gr{lf.ioul('nl IJ:lJlOunl.': oj I'cm:ling
sulistances have been (:(I1I1,/JilU'fl as deuuulr!ed by the clrnaiio1l.. In
pl'n,etin(~ Uw tiLmtioll is stuPlln(l at It lll'l1r:tic:al point, krnwri the
enrl point, wlwn t.liu rnaetioll is shown to ho C:()lll}Jlr~tr, by the
c:lmllge in culm of a TllutienhLr indicator or I)y ::-:OI11e O1,her device.
In every titration tlw PIl(I poi Ilt. ancl s(,nidliometri(~ point nn.lst
eoineicln vury eln;-;nly if th(~ rmmlt iH to I)c of value ill unalyti(:al
work.
Before 1llHlortnkillg HllnlYI,(IH by nwtllOds of aeiclillletl'Y and
alkalinlOtl'Y, Lhn st.llckIlt shoultll)(H~()lD(: l'ulllilit1l' wit h t.he t]wol'eti-
enl eOllsidernti()]lH illVnlv(~cl.
Titer.----The tiLer of a HollltioJl is t.lw llllllllwr (rr grams oj' solute
(~()ntaillml in 1 (~(\., 01' tho ivei~ht of [Lny sllllHtf1l1C'O which will
rmwt with 01' )Jp (IXl1et!y pquivalent to 1 e(~. or tho s()lutioll, 1'(1,
a solutioll eODtrLilling; :Hi.47 Om. of .Hel in 1,000 (?(:. has an Hel
titer' of 0.0::;(;47. Tlw NaOn tit.er of Lhis Holut,ioll lllay be found
from the rdtdiioJlshi)J:

Hel :NaOH::0.0:3647:x

where x is the wuight ill gl'ams of Plll'C NaOH whieh can be

neutralized by 1 ee. of t.1)(' above :wid aUlI is cqUttl to 0.0400 Gm
66 QUAN'l'I'1'ATIVE PIIARMAC]!,'UTICAL CIlEMISTllY

In like mmmer, the desired titer of :1ny other substance may be

found by direct proportion.
Theory.-----The 11ydrogcm ion (:olleentratioll inflncmemJ most
reactions employed in ltnulytic[Ll methods WiH\tiwl" they are
lleutralizlLtion, pn:eipital;iou, or oxidntion-n:dlwtioll reactiolls.
It Ims bew establishecl that in any aqueous solutioll there aw
always hydrogen and hydroxyl iOllS prci-lcmt nnel that wluLtevo)"
the concentration of the one, the cOIlecutratiOll of thn othe)"
is sueh that the product of their eoncentrationR is :1 eOllRLanL fm
any given tempemture.
[I-P] X [OH-] = a constant for any given temperature, and
in pure water at about 22(;., [I--II-] X [OH-] = 1 x 10-H. Sinen
thiR relationship is true, the eoneentratioll of hydrogen ions II-I+j
exprf~ssl\d in terms of gram-ions per litor in pure wfLter is equtLI
to the square root of 1 X 10-H '= 1 X 10-7 (O.O()()OOOl), and tile
hydroxyl ion concentration [OH-] al:'-io is equal to :I. X 10-7.
The degree of acidity or alkalinity of a solutioll (:al1, tiwrdorn,
be defined in tenni; of its eonc~(mtration of hydrogen ion or
hydroxyl ion, sinec when the value of one is known tho vnhw of
the other ean he efLlclllatcd; 1.e., if [IP] = 10--r" thcm [OH--] =
1O-!1, Rince [H+] X [OH-] mURt equal 10- 14 Solutions which
have a hydrog(Hl ion coneentmtioll greater than 10"-'/ an~ udelie,
and those that h[we EL hydrogen ion concentration lnsR than 10- 7
are alkaline. Thus [L Roilltion of 0.01 N Hel eontaitlR_ 1(}-!~
Gm.-iolls of hydrogen per liter, assuming complete ciiHsoeint.ion,
and is twid in rerwtion tLlthongh it eonbins 10-- 12 Gm.-ions of Oll-.
To avoid the disadvantage of employing n. dual HYHtmn of
definition, it is enstomary to use only the hydrogcm ion eoneel1-
tmtion to ehamdorize l1 Rolution, wlwtlwr it 1>0 aeidk, nClltml, or
alkaline, the hydroxyl inn noneontmtioll }Jping impli<\cl.
pH Value.--In a(lditioll to the oxpcmelltial fonn of (\Xpl"(~Rsinl-!:
hydrogen ion eoneentratioJl nmployecl above, allOthe:r systnrn
suggested by Si_irellsen is ill common usc. III tlw lattm' Rystrlll)
only tho oxpouent io, llHed with itR negn,tive sign dropped and thH
symbol pH prefixed, c.g.:

[H+] lO-z.Of> then pH = 2.06 acidic:

=
[H+] =
10-- 7 tlwn pH =' 7 neutral
[H+] = 10-1~.~r, then pH = 12.9.5 alkaline
 P1UNCIPLES OF VOLUMETRIC ANALYSIS 67
pH is genendly definod as the Zogan:thm of the reciprocal of the
. emwen l ra t'um, wntten
}L1JI1:rofwn wn .' 1
pH = log [H:'T Since the
hydrogell ion coneollLmLion of :1 solution i" a fraction denoting
the nUlllhnl' of gnwL'\ of llyrirogeJl ion cOlltnined ill n litcr, it call
be (~xpn~:-;H(!(l as l/IH+I which i8 the reciprocal of [IF1. By
tlw usn oj' the l'oc:iPl'u(:ltl, tho negative exponent is avoided; thus,
1/101 iH Llw Ramo :ts 1 X 10-.1.
The fae!; that it :-Julution eontniu8 O.()OOl Om. of hydrogen ions
per liter may, tlwrdore, b() exprel,:-;pd in vario1ls ways as follows:
111+) = 0.0001 Gill. pOl' liter
[H+] = 1 X 10-'1
log [H+] = - 4
-. log 111-'] = ,~
1
log; [H'j '-7' 4
pH = 4

Cone()lltml;iol1s thut ILl'e llIlCvml decimal fl'lwtiolls can also be

expl'PRIw(1 ill t,nnlUi of pH lmits; e.g., if the hy(lrogoll ion f:ollcen-
trntioll of :1 soluCioll is 2.73 X 10-.1 the pH vaIuD js 3.5M. By
l'Oi'eronc:e to [t lognrithm tabk, it if:! fOllud that log 2.73 = + 0.436
lind log 10-.1 = - 4.0()O. Since the lllllllbers [11'0 to be multiplied,
t1w \Op;H ftl'(\ [1ddctl and -4.000 + 0.436 = - 3.564. Therefol'e
log [H+] = -:~.5(i'1, -log [IF] = a.564:, and pH = 3.564.
COllvnl'cwly, whnn tlw pH is given, the hYllrogen lOll eOlwentra-
tinn enll jJP (:(delllt~tl\ll. Thus pH = 9.63 Illoans that [I-I+] =
1 X 10 \1.1;:1. The eXpolKmt -9.6:3 = -10 + 0.37, t1ud 10- 0. 63
= 10-- 1(1 X 10+(1;11. From Lhe logarithm CahIfl, it iH found
thnt the expouent O.3kfCOI'l'(,:;pondR to the Humber 2.3"1. There-
fore [H'l = 10- il . lla is eqllal tu III-Ir = 2.:34 X 10- 10 .
Two ll]'[l('.tienl methods of determining when the hydrogen
ion c:oJl.(:eut.mtioll in (1 floluLioll has acquired n, eertain value have
been (1(wf11op!:d, lltunely, the potentiometric: method, and the
f,()lorimntri(: lll()thod (Bee page 272).
Buffers.,,--ln many Hlwlytical pI'OCedureR a ehnngc in the
hydl'Ogen ion eorwentrntioll oecuI'S [18 a result of chemical reae~
tiOll. In certain eru;es, t.his ehange in the concentrat.ion of hydro-
gen ion may prevent the reaction from going to completion
68 (JUfiN'l'1'l'A'l'IVlE PlIARMACIW'l'IC'AL CllRJJHS'l'RY

qU[l,ntita,tivoly. Since the chango in hydrogen iOll concentration

is caused by the formation or (wid Ol' hase in tlw eoul'se of the
re::wtion, somo l)]'ovisioll mllst he Hl:tc[e 1;0 eliminate the acid or
base forme(t in slU,.ll :1 lIl:UllJe]' as to lilttinbtin n H(msibly eonstnnt
hydrogen iOIl (~(m('.()lItl'::t(,iOll. ThiN iN HHllally ::weolllpliNhecl by
lnemlN 0(' n lmffpl. TIl(: tl~l'm ]Juffer iN applied alike to. tlw sn,)ts
of w(~ak. [l.eicls [tnt! tu tIl<: sa.!Ls of wPlLk. l>:wnH. Soln tions of tlWS(~
,;dts l'CNi"t, dml\g(~ ill hytlrogen ion (~one(m[,l':d,ion on the :\,(I<1i\,i01l
of HIlI[I,Il flllJOllll LH uf cithor :wid 0]' h[LH(~. In the titration of
acetie neid wiLh Hodium hyclroxidn, thn sodinm acelintu fOl'nwd
acts as a IlltiTel' to prevent ::t Ittrge eh:wge ill liyrlrogoLl iOll (~()ll
eentration. Anctil: a.eill in aquculls solutioll iH slightly dissoeiaLed
a:o :obOWH ill tIw followillg; (~q\latiollH:

In readion (1), the slighLly dis:o()(~iaLe(l acetic acid forms H+

and CH:1COO-- iOllS which combine with thfl OH- all (1 Na l
ions from equation (2) to form HOH ill equatiou (a) nlld CH:1-
COONa ill equation (4). The sodium [wctnLo lH highly (lis-
Roeiat,ed into N!t+ awl CHaCO(Y iOlIR. 'I'hi:-; results in It high
cOllcentmtion of CHaCOO- iOllH in Lhe sulution ltlld repression
of the H+ iOlL concentration fOl'eing pqtmtion (1) toward tho left.
As [tc1di.tiom~l qtULntities of NaOn m'e t1Xld(\d, tllP :n+ ion i:-;
removed as HOH and fmther RInltll 1Ll1l0lllltH of ncntie aeid clis-
soci[1,t,(:l so that a eontinuolli:l Hupply of H+ lOll is provided. Thn
solution will continuo to contain an (~X(l()RH of HI- ion until all of
the CHaCOOH has been eOllsumed. Tho addition of a small
(lUulltity of nllmli in exceSH of tho l'oqllirect arnount will thou
cause the OH-- iou eoneen(;r:d;ioll to exceocl tho H+ ion eorwentm-
tion and the solution will react, alkaline.
Tho prineiple of buffer u(:tion is often employed in l1llnJytieal
moth ods in whieh acicl or base is generatnd as a result of the
alU11yt,lt:111 reaction. In such cases, tho additioll of an amount of
buffer "alt rel:1tively large with respect to the amount of acid or
 l>lUNCIP/;}~8 (}fI' F()LUM1~'1'lU(} ANALYSIS 69

basu gmlO.mted (iaUHer,; n, frrwtional diminution of Lhe buffer ion,

through .itH nnion lVith tho hydrogen iOll to forIn the slightly
(lisso(:iatc:d twid (:ol'l'nSpOllclilllJ; t(l the llll.l'fm cinlt employed. The
dillliulltioll ill t,]u: :MliOllllL ()f hufTpl' mdt will Llwn bn so slight Lhat
j.Jw hydl'()gnll iOll ('()lI(:nntl'atioll will rema.in pl'aetic:nlly constant.
In iodilllld,l'l(: aHsays wIiP]'(: rn iN fOl'llWd a:-; It lll'ochwt of the reac-
Li()n, H()diultl Ilit:a!'])(IW11.n is w;(.(l rtH n. Ind'ft,l' to maiutain fLll
npPI'OXilllrddy (:lllmi.n,11L llYlll'ogml. ion e(me('lltration.
N eutralizatiol1 Reactions. -It is importl1llt t(J remember th[Lt,
ill tC'.l'Ins (Ii' L1u: 10Ili(: 1,II('o1'Y, n, IlPlltrnl ::;oliltion eontains H+ an.d
o Fl" JOIlH ill L1lu t':l.IIlt: ('Olli:on tl'11l.ioll ILOl in pitre water, namely,
O.O()()O()()l UIlI."j()1I 1)(:1' liler ()f Cl()lution. AllY sllhst.anee which
I:all fll],lIi~;]1 I'X('I','4,'-\ 111 jIJIlS wll.lll1 ;]i'~";Hlvl!d in water i.e.; aeidic and
allY snhH(:LIH:t: which \::tll i'Ul'lJiClh I'x\:m;:j ()H- jom; when diflflolved
in ",atlll' iH blt,~il:.
Fl'Olll (.111" poillt; of view of the iOllic theory,
i,; Lhe ILnion Ill' n+ ions from un twirl with
(']WI'(:i'OI'(\, 'linlll)'{//.i:~(/,li(}1/.
(HI- illltH frOlll a haHn, {;o forlll 1I1()1(:(:ltl(~S of watur. Tho other
l()U"! fllJ'llislwtl I),Y t1w add. 01' klS<1 mayor may not rol1ulin

;:oltil,ill!'Ii.
Nt:\ILJ':I.lizn,Lilill l'(:lL(:1.iIllIOl lllllS!; pl'oec;:d j.o eompletioll to Le of
valJl(: ill qlJ:l,IlLital,ivu :LlIaIYHiH. H.eaetiOllK lllay be eaused to go
(,1) l:umpld,i01I ill lL 11111111)(;1' of way::;, 'i.e., by tlm formatiun of n.
Hlig;lttly diH:-ml:ia{,(:d fmll:,;i;nnt:e [1tl :1 reaction product; by the
rt:l1luvnl or Ollt: OJ' mort: uf 1.1IU pl'o(lude; of the reCLctioll as a gas;
by the 1'(:lIlOval of (HU' 01' III 0],(: of tilO pnllluets of relletion as a
]lI'PI:ipi (.:Lk; nlill hy lI.rldint~ an eXt:f'''~ of olle of the r.eaetl1nt.s.
'1'11111', il' 1lCa :1IH1 NaOn ;Ll'() cliHiiolved separately in water to
Ilwkf'. dilul.n afjll('()\IH Nol111.iollH, nn.c:h kuhstanee iN almost eom-
plntdy (liHHoei:L1,nd into i()IJ:, [l'OlLctiollS (1) ml(l (2)].

Upon mixing tho two solutiollfl somo Na+ and Cl- ions unite
forming; NaCI l1Io1eellh's-ronetinn (4). Wn,ter is only slightly
70 QUANTITATIYE PHARMACEUTICAL CIlEMIS'l'RY

ionized, so neaTly all of tho H+ ions and OH- ions unite to form
HOIl molecules-reaction (3). The removal of H+ and OH-
ions ill. reaction (3) causes reactions (1) and (2) to proceed to
completion, so that if equivalent quantities of HCl and NaOR
are employed, the final fJolution will contain only HOH, a small
qnantity of ullclisRocilttccl N aCl molecules and N a+ and Cl- ions.
If one of the produets of a reaction ifl removed the reaction
procecds to completion readily; e.g., when KN0 3 in solution is
treated with Hel solution the equilihria represented by equations
(1) ancl (2) exist prior to their admixture.

(1)
KN03~ ~K+ +N03
(2) + +
HCl Cl-- +H+
11 (3) i1 (4)
KCl HNO a

,Vhen the two solutions are mixed, equilibria become estab-

lished between the four reactions represented. If the solution
is boiled, RNO:l is volatilized as fast as it is formed in reaction (4),
removing H+ and NOa- ions, thereby causing reactions (1) and
(2) to go to completion. Upon evapol':1tion of all of the water,
KCl rema,ins.
Unclcr Precipitation Iv[ethocls numerous examples will be
eneounterecl of the use of an excess of one component of a reaction
and of the removal of ono of the products in the form of an insolu-
hIe precipitate as methods of forcing reactions to completion.
Indicators.-The indicators commonly used to determine the
enc1 point in neutralization pl'Oeesses are as a rule complex
organic compounds eallable of existing in two forms of ditlorellt
color which arc mutually convertible, one into the other, at
given H+ and OR-ion coneentrations. Three theorie8 have
been proposed to explain the change in color of indicators which,
briefly stated, arc as follows:
1. The phy:-;ico-chcmical theory attributes the color to certain
ions an increase of which causes the appearance of a new color,
and the decrease of which causes the disappearance of a color or
the appearance of a different color.
 PRINCIPLES OF VOLUMETRIC ANALY8IS 71

2. The organic thcory attributes the color of illdicntorR to

certain groupings of the elements in [1 componnd, fLlld the chungtl
in color to change in molecular structure.
3. The colloidul thcory aSSUlllPS that indielltors f01'I1l colloidal
solutions the change in color of which is dnpCllclent upon change ill
size of the colloidal particle.
The point at which an indicator changes color in any given
. titration is dependent all the hydrogen ion concentrntion of the
solution and may not be indicative of the ahsolute neutrality
or completion of a reaction. The ac(~olllpanyillg tahle of the
eommonly used indicators gives the H+ ion concentration~
expre!3sed in terms of pH between which color changes occur.

TABLE VII.-COMMONLY USJW INDIC'ATons

Color
Illcliclt tor
Aeirl Alkaline

Bromcresol purplfJ ...... . 5.2 to 0.8 Yellol\' Purple

Bromphenol blue .............. . 3.0to4.1i tBlIow BIuisl] violet
Bl'Omthymol bluc .............. . li.Oto7.6 Yellow lllue
Cochineal ..................... . 4.8toG.2 Yellow Lilac pink
Congo red .................... . 3.0 to 5.0 Blue Reddish yellow
Litmus .................. , .... . 4.5 to 8.3 Red Blue
Methyl orange ................ . 3.1to4.4 Pink Yellow
Methyl red ................... . 4.2 to 6.3 Red Yellow
Phenol rcd .................... . 6.8 to 8.4 YnUow Hed
PhenolpMhnlein ............... . 8.0 to 9.8 Colorless Red
Thymol hlue .................. . 8.0 to 9.(\ Yellow Blue

Thi" tt1ble shows tlmt methyl Ol':1nge exhibits its acid color,
pink, at a pH of 3.1 and its alkaline color, yellow, at a pH of 4.4,
while bfltween theHe pH values the ('0101' undergoes trallsition
from one shade to the other. At a pH of 4.4 the solution is
Rlightly acid, since at neutrality pH = 7.
It would appear that an indicator which changes eolor exactly
at the neutral point, pH = 7, woulcl.be required in every case,
but this is not true. When a strong acid, sueh as hydrochloric
acid, is titrated with a strong base, like sodium hydroxide, the
change in concentration of hydrogen ion becomes very rapid as
72 QUA.NTITATIVliJ PHARMACEUTICAL CHEMISTRY

the stoichiometric. point is approached so that very smnJl amounts

of acid or alkali in excess carry the concentration of hydrogen
ion far to one siele of the neutral point, and the indicator will
give a sharp end point. If a solution of 25 ce. of 0.1 N HCI
is titrated with a solution of 0.1 N NaOH, the equivalent point
or stoic:hiometric point. would be rcached when 25 ee. of 0.1 N
NaOH had been added. Theoretically, the solution at the end
of titration should })e identical in eomposition with a solution
prepared by dissolving a known weight of sodium phloricle in
the calculated amount of solvent. ,,yhen methyl orange is llsed
as indicator, the end point, as shown by change of color, app(~ars
when about 24.95 ce. of 0.1 N NaOH have heen added or while the
solution is still slightly aciclie, but if phenolIlhthalein is used as
indicator, the emI point appears when about 25.05 eo. of 0.1 N
NaOH haye been added or when the solution is slightly alkaline.
The deviation of the end point from the stoichiometric point is
so slight that it is negligihle in this ease.
'When a weak add is titrntr~d with a strong hase, tho change
in hydrogen inn coneentration ill the vicinity of the stoichio-
metric point is not so abrupt; e.g., if 25 ec. ci'f 0.1 N acetic aeid are
titrated with 0.1 N NaOH, 2fi ec. of the alkali should be required
to attain the stoid1iOlnetric point. When methyl orange is
employed as inclieator, the end point is reached when about
17.5 ce. of 0.1 N NaOH have bcpn added, hut if phenolphthalein
is used the end point is reached when practically 25 ee. of 0.1 N
NaOH have been added. Ohviously methyl orange would not
be a satisfactory inclicn,tor in the above case.
Acetic acid titrated with sodium hydroxide forms sodium
acetate: CH 3 COC)H + NrtOH CIM~OONa + HOll. The
acetate ions. from the highly ionized sodium acetatc foree the
reaction to the left CH3COOH<--~H+ +. CH 3COO- before all
equivnJent qmmtity of NaOH has been added, and thc hydro-
gen ion. concentration becomes very sIllall before all of the hydro-
gen of acetic acid has been replaced. In this case methyl orange
which changes color nt a relatively high hydrogen ion concentra-
tion 10'-4..1 will show alkaline color gradually long before the
equivalent quantity of alkali has heen added, and the end point
will not be sharp. Phenolphthalein, however, does not change
color until the true neutral point pH = 7 has been passed, and
 PRINCIPLES OF FOLUMETRICANALY8IS 73

a drop of alkali will than reduce the concentration below [H+l ==

1O- D 8 or pH =-= 9.8 where phenolphthalein exhibits a change of
color.
A weak base titmtecl by a strong ll,cicl, c.g., :llnmoniUITl hydrox-
ide titmtod with sulfuric 01' hydrochlori(, [will, exhibits tIte
r()\"cniC of the above ('ondition, tiS would h" expeeterl, and methyl
orange is a suitable indicator while phenolphthalein is nut..
Rules for the Use of Indicators.-The following rules should
he observed in t.he use of indicat.ors:
1. Use two drops of indicator solution for a titratioll uules:'1
otherwise directed.
2. When a strong acid is t.itrated with a strong alkali or a ::;trong
alkali with 11 strong acid, either methyl omnge, methyl red, or
phenolphthalein may be uRed.
3. When a weak acid is t.itrated with a strong alkali, use phenol-
phthalein as inclic:ator.
4. ~Whcn a weak alkali is titmtecl with a strong ncid, use methyl
orange ns indicntor.
5. A weak alkali should never be titrated with a weak acid or
vice versa, since no indicator will give a sharp end point.
6. The appearance of a color is more easily observable than is
the disappearance. Always titrate where possible, therefore, to
the appearance of a c0101'.
The criteria of a good indicator for an::dytical purposes are
that there shall be a sharp contraRt between the two eolol's which
it exhibitR in add and alkaline media anel that the change in
color shall take place over a very small range of hydrogen ion
concentrati.on. Unless otherwise stated, each indicator solution
should be so adjusted that when 0.15 cc. of the indicator solution
is added to 25 ce. of distilled water, 0.25 cc. of 0.2 N add 01' alkali,
respectively, will develop the characteristic c,olor changes.
Some Commonly Used Indicator Solutions.
Bromcresol Purple. Di-bl'omo-o-cresol-sulion-ph thalein.-A
solution of 0.1 Gm. of the dry powder ill 9.5 cc. of 0.050 N
NaOH diluted to 250 ee. is used as indicator solution, Two to
four drops of indicator pel' 100 cc. of solution should be used in
titration unless otherwise directed.
Bromphenol Blue. Tetra-brolllO-pheuQl-sulfon-phtha,lein.--A
solution of 0.10 GIll. of dry powder in 100 eo. of 50 pel' cent ~
74 QUANTITi1TIVE PHARMACEUTICAL CHEMISTRY

alcohol is used a~ indicator. Foul' to six: drops of indicator per

100 ce. of solution should be used in titrations.
Bl'omthymol Blue. Di - bromo -thymol-sulfon-phthalein.-A
solution of 0.10 Gm. of dry powder in 100 ec. of 50 per cent alcohol
is employed as indicator. Two to four drops of indicator solution
per 100 ce. nre ufiecl in titrations.
C(Jchineal.-Cochilleal consists of the dried female imlcct
Coccus cacti. Th(~ indicator constituent is carminic acid.
Cochineal indicator Holntion is prepared by macerating 1 Gm. of
the puwdered, dry insc('t,s for three days with 25 per cent alcohol
and filtering, In titratiollS, 2 to 4 drops of indicator solution
should be used for eaeh 100 cC. of titration mixture unless other-
wise directed.
This indicator iH rmitable for the titration of alkalies and alkali
('tLl'bOllntes, ammonia, and mallY alkaloids and inorganic acids,
but it should not be used for the titration of organic adds.
Congo Reel. Diphcnyl-diK:lzo-bis-a-uaphthyl-amine-4-sulfonic
Aeid.--The indicator solution if; prepared by dissolving 50 mg.
of the dye in 100 cC. of 70 per cent alcohol. This indicator is
used in the Kjeldahl ammouia determination. In titrations,
;3 to 4 drops of indicator are used for each 100 cC. of solution.
Lit'lntts, or tournesol, ('ollsists of a blne pigrnent .obtained from
various Hpecies of Roccclla De Candolle by fermentation. The
pigment mixed with chalk 01' other diluent is marketed in eubes or
fragments. The blue coloring matter consists chiefly of azolit-
min. The indicator solution is prepared by extracting 25 Gm. of
the Holicl cOlnmereial product with three suecessive portions of
100 ceo of boiling alcohol to remove red coloring matter. The
undissolved residue is extracted with eold distilled water to
remove soluble carbonates. TheHe extractives are discarded.
The residue is theu treated with 125 cC. of distilled water, boiled,
cooled, ancl filtered. The filtmte constitutes the indicator
solution.
Other indicator solutions are superior to litmus for titration
purposes, but litmus test solution serves well for rough qualitative
tests of hydrogen ion coneentration.
MethylOra.nge. The Sodium Salt of Dimethylamino-azo-ben-
zene Sulfonic Aeid.-The indieator solution is prepared by
r dissolving 0.10 Om. of the dry salt in 100 cC. of distilled water,
 PRINCIPLES OF VOLUMETRIC ANALYSIS 75

Two to four drops pel' 100 cc. of reaction solution arc employee!
in titrations.
The color change is very sharp if too much indieator is not
used. Methyl orange is used frequently in the titration of strong
acids and st.rong alkalies and especially in the titratioll of -Weak
bases, e.g., NH 4 0H. It is also a good indicator to use in the
titration of the salts of weak acids sueh as carbollateH, bomte~,
:-mlfides, etc., with a strong ac:id, sin('e in t.hese CUf'(1S the adds
liberated in the titration reaction arc Loo little ionized to affed
the indicator. It does not give tJw proper end point in the titra-
tion of alkaloidl-l or organic acid8. Thi!:i incli('atol' should ne\'pr
be employed in the titration of alcoholic soluLion8, hot solutions,
or very dilute solutions.
IJ![ ethyl Red. o-Cal'boxy-benzenc-azo-dintetbyl-aniline.---The
indicator solution is prepared by dissolving 0.10 Om. of the dry
dye in 100 ce. of 95 pel' cent alcohol. Two to four drop;,; of the
reclUlting solution are used per 100 (~C. in titrations. 1\;1 ethyl
red indicator is especially useful in titmting ammonia, wpak
bases, and alkaloids, but it is 110t suitable for the titration of
weak organic acids,
Phenol Red. Phenol-sulfon-phthalein.-The indicator solu-
tion is prepared by triturating 100 mg. of the dry powder in n.
mortar with 14.5 ce. of 0.020 N NaOH until solution is ('omplete
and diluting the resulting soilltion to 250 ceo wit.h recently boiled
distilled water. After the removal of any prccipitl1,te formed
upon dilution, the solution is ready for usc. In titrations, 3 to
6 drops of this indieator Rhould be used for eael! 100 ec. of
solution.
Phenolphthalein indicator solution is prepared by dissolving
1 Gm. of phenolphthalein in 100 ce. of 95 per centakohol. Two
to three drops of this indicator should be u8cd per 100 cc. of solu-
tion in titratiol1R lln1ess othcrwiRe dirc('ted.
Phenolphthalein is an excellent indicntor to use in the titra-
tion of wenk organic and inorganiC' acids, most alkalies, and alkali
salts, but it is not satisfactory for use in the titration of ammo-
nia, alkaloids, 01' cold solutions of carbonates and bicarbonates.
It functions well in akoholic so~utions.
Thymol Blue. Thymol-sulfon-phthalein.-The indicator solu-
tion is prepared by dissolvillg 40 mg. of the dry substance in
76 QUAN'I'ITATIFE PllARM11CEUTWAL CHEl!.[I8Tl~Y

100 cc. of alcohol. Two to four drops of indicator should be

employccl in titrations per 100 cc. of reaetion solution.
Sensitivity of Indicator Solutions.-Indicator solutions ShOllld
be tested for sensitiveness. This is dOlle by adding 0.15 ce. of
indicator solution to 25 ce. of distilled water and titrating the
rmmlting solution with 0.02 N alkali or aciel. Upon the addition
of 0.25 cc. of the standard alkali 01' 1],('id solution, tho respective
cbaraeteristie color should develop.
Questions and Problems
1. Defiue the foll(nl"ing; tprmfl: (II) stoichiometrio point, (h) end point in
titmtioll, Cc) Illdie:Ltor, (d) buffer.
2. If it iiolutioll has a hydrogen ion conecntr[l,tion of 10- D.", Wh:Lt is its
hydroxyl ion concentration?
3. In a solution having a hydrogrll ion concentration of [IFJ = 1O- r. 5,
Wh~Lt woulrl be tho pH value? Would the solution be acid or alkaline?
4. What indicator iR lwst suited to the following titnLtions? 'VlIy?
(a) alkali cal'bon:1te wit h s\11hll'i() twill, (li) aectic twirl with sodium hydroxide,
(c) alYlmonillJU hyclroxi(l<l with bydroehlorie fwill, (d) org:min acids \"ith
sodium hydroxide.
5. How nULY the scnsit,lvrness of an inrlimtor he determined?
6. Why is methyl orange n good j'ndi(:n[,or to usc in tlm titratioll of
NH 1 0H'?

Standard Solutions.-The coneentration of standard solutions

ca.n be expressed in various ways as follows:
A normal solutl:on is one which contains 10m. equivalent
weight of reagent dissolved ill sufficient solvent to make 1,000 ec.
of solution.
Ala/at solutions are those which contain ,in 1,000 ee. a gm1J1-
molecule equ.ivalent of the 1'eagent, irrespective of the valency
of the molecule. Thus, a molar solution of sulfuric acid would
contain 98.08 Gm. of 11 2SO,1 in a liter of the solution, !Lnd a molar
p,01ution of potassium dichromate would cont.ain 2H4,21 Om. of
K 2 Cr Z07 in a liter of the solution. Solutions eontaining in
1,000 ce. one-tenth of a gram-molecule of the reagent are desig-
nated "tenth-111obr," M/10, and other molarities are similarly
indieated.
A molal solution is one which contains 1 mole 01' Gm.-moleculll,!,
weight of reagent dissolved in 1,000 Om. of solvent.
The coneentratioll of official standard solutions, volumetric
solutions, is expressed in terms of normality or molarity. It is
 PRINCIPLES OF' VOLUMETRIC ANALYB18 77

very important that what is meant by a normal and a molal'

solution be clearly understood. A normal solution contains one
gram equivalent wwight of reagent per liter of Sol1lt'ion, i.e., that
quantity of reagent whieh is chemieally equivalent to S.OOO Om.
of oxygen or 1.0078 Om. of hydrogen per 1,000 ceo The gram
equivalent weight, in neutralizatiofi reactiowl, is equal to the
gram-molecular weight (molecular weight of suostanee in grams)
divided by the number of H+ or OH- ions entering irtto or formed
in the reaction. Fr()lll the above definition of [l, normal solution,
it is obvious that a llorIllal acid solution eontains 1.0078 Gm. of
available hydrogen ion per liter; thus, a llUl'ma! solution of
hydrochloric: aeid contains 1 mole, a6.47 Gill. of Hel, and a
normal solntion of sulfuric [Lcid l'ontains (L5 mole, 49,04 Gm. of
H 2S0 4, pel' 1,000 ceo 01' solution, beef111Se tho~,e weights of the
respective acid:;; contain 1.0078 Gm. of availahle hydrogen. A
normal alkali solution contains sufficient available hydroxyl
ion in 1,000 eC. to nombine wiLh 1.0078 Gm. of hydrogen ion or
17.0078 Om. c.(j., 1 mole, 40.00 Gm. of sodium hydroxide or
0.5 mole, 157.75 Om. of barium hydroxide per 1,000 ec. of solu-
tion. A normal oxidizing solution has an oxidizing value of
8.000 Gm. of oxygen equivalent to 1.0078 Om. of hydrogen
per 1,000 cc.; i.c., 2KMu04 used as an oxidizing agent gives up
5 atoms of available oxygen equivalent to 10 aeoms of available
hydrogen, thorefore a normal solittion will contain 0.2 mole,
31.606 Gm. of KMn04, per 1,000 cC. of solution. It is evident
that. equal volumes of normal alkali and acid solution when mixed
will exactly neutralize one another if they react quantitatively
and that equal volumes of normal oxidizing and reducing solu
tions will exactly use up one another. .
The letter N is used to indicate normality. The normality of
a solution may be represented in two ways; i.e., a solution of
exactly normal strength is indicated by 1 Nor 1/N (normality =
1.0000), a double normal strength by 2 N 01' 21N (normal-
ity = 2.0000), a half 01' seminormal by 0.5 N 01' N /2 (normal-
ity = 0.5000), a clecinormal by 0.1 or N /10 (normality =
0.1000), and a centinormal by 0.01 or N 1100 (normality =
0.0100), etc.
In practical analytical work, it is nnneCE)ssary to prepare or
maintain solutions exactly normal or tenth-normal, etc. It is
78 QUANTITATIYE PIIAIlM.t1CEU'l'ICAL CHEMISTRY

usuflJly ettsier and just as satisfactory to determine the normality

of a solution which closely approaches the value of an exactly
norlllal solution. The relationship of an approximately normal
solution to an exactly normal solution is expressed by a factor,
often called the normality factor; e.g., a normal solution of sodium
hydroxide contains 40.00 Gm. (the gram equivalent weight)
per 1,000 ee., and an approximately normal solution contains
40.42 Gm. pel' 1,000 ee. The latter is then40.42j40.00 = 1.0105
normal and [IllY number of cubic centimeters of this solution used
can be expressed in terms of exact normality by multiplying by
this factor; thus 20.1 ee. represents 20.1 X 1.0105 = 20.31 cc. of
1 N Rolution. The standard alkali solution should be labeled
1.0105 N NaOH. The normality factor expresses the ratio
between the weight of reagent present in a liter of solution and
the weight that would be present if the solution were exactly
normal. Usually the factors of volumetric solutions are limited
to four significant figures.
'fhe stf1ndard acid i'lolutions used in acidimetry and alka-
limetry [Lre usually prepared from hydrochloric or sulfuric
acid. Bither aeid may be used in most titrations, but hydro-
chloric is preferable to sulfuric acid in the titration of com-
pounds whieh yield a precipitate with the latter, such as barium
hydroxide, and sulfuric acid is preferable in hot titrations,
since there would be danger of loss of hydroehloric acid due to
vola tilization.
Tho standard alkali solutions commonly used are sodium
hydroxide, potassiulll hydroxide, and barium hydroxide. These
hydroxide solutiolls absorb carbon dioxide from the air, thereby
changing rapidly in concentration. Sodium and potassium
hydroxides may beeome eontaminated with cm'bonates which
impair their usefulnoss due to thc liberation of carbon dioxide
during acid-alkali titration. Barium hydroxide solutions remain
free from carbonates, si1l(~e absorbed carbon dioxide is precipi-
tated as insoluble barium carbonate; this, however, decreases
the concentration of barium hydroxide in the solution. Alkali
solutions should be prepared carbonate-free, and they should be
protected from carbon dioxide by means of a soda-lime absorp-
tion tube. All alkali solutions shonld be restandardized fre-
quently.
 P1UNCIPLES OF VOLUMETRIC ANALF8IS i9

Exercise 10
Preparation and Standardization of Hydrochloric Acid Solu-
tion.-N o1'mal "hydrochloric acid solution. may be st::.mdardized
against pure sodium. (~arhollatp, standard solution of :-iodium
hydroxide, or gravimetrieally by precipitating and '\'Highing the
chloride ion as silver chloride.
Object.-To Prepare and Standardize Normal IIydroehloric
Acid.
Materials Required.-Hydl'ochloric arid.
4 Gm. of anhydrous sodium eal'bollltte.
Procedure.-l. Dilute 95 cc. of hycll'or:hlol'ic acid wilh snflil'iellj, (li.~lilletl
water to make 1,000 ~(l., lend mix thor<\\\ghly.

Since hydroehloric acid, specific gravity l.li5, {'OIltnins about

36 per cent of Hel, the l'c:-;ulting solution :-;hould be slightly
stronger than normal. Thus, 95 X l.li5 X 0.:16 = "10.18 Gm.
HOI. Since this quantity of He1 is dissol\"{~d ill 1,000 ce., the
solution should conta,in approximately, 40 Gm. He\.
2. Weigh about 4 Gm. of pure sodilllll earhollatl1 on a rough halance and
place the salt in a porcelain dish of suit!lble ellpRcity. nc'at the .~[ll'bOlmtl1
for 1 hr. over a low BUllscnllnmH or in an OYl1Jl at. 270"(;. rlllllll.llo,, it to eool
in a desiccator. Weigh accurat.ely t.wo portion~ of about 1 E (1m. c(l!'h of thl1
cool dry salt, recording the weights.

Although pure sodium carbonate can bo pureha~ecl, t.he Halt

contains varying amonnts of water of erystallization and llloiR-
ture. When it is dried, the absorbed moisture is dl'in'n on
as well as any water of crystallization, since sodium carbOllllb
becomes anhydrous when heated to lOaDe.
3. Transfer each of the samples to 250 cc. ErlerllJlcynr flusks and proeeul
to titrate them separately 1IF; follows: Add llhout IOO ce. of distilled water
and 2 drops of methyl orange T.S. aud slulke until the carbnnMe is eODl-
pletely dissolved. Fill the glllss-stopperpd burette eX:1clly to the mark
with the acid solution and rUll acid slowly into the flask with continuous
shaking until the solution just becomes pink ill color. A white porcelain
plate or a piece of white paper placed beneath i,he flask renders t,he end
point more easily discernible. Wash down the iIlsitie wnliH of the flask
with distilled water from it wash bottle, ami if the solution becomes yenow,
add acid drop by drop until the faint pink color is restored, taking care that
the last drop which adhered to the b\ll'otte tip is introduced into the solution.
80 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

When titration is c:omplete, read the burette accurately and record the
volume of acid used in the titration.
Assuming that a sample of sodium carbonate weighing 1.6250
Gm. required 30.20 cc. of acid in the titration, the calculationH
are made as follows:
If the acid were exactly normal, aeeording to the definition of
. . 1 mole Na"CO a
a normal solutIOn, 1,000 ec. would neutralIze 2 w =

10~.99 = 53 Gm. sodium carbonate, and the sample used would

require:
53: 1,000: : 1.6250:x
1625.0
x = -53 = 30.66 ec.

Since only 30.20 ce. of acid were required in the titration, it is

evident that the acid solution is ~tI'Onger than normal, and the
factor is 30.66/30.20 = 1.0152 N. The bottle eontaining the
acid should be labeled 1.0152 N Hel.
The calculation may ah;o be made by the method of equiva-
lents, namely,
Na 2 CO a + 2HCl = 2NaCl + H 2 0 + CO 2
2)105.99 2)72.94
53 Om. 36.47 = 1,000 ee. N HCI
0.053 Om. Na 2 C0 3 = 1 cc. of N HCI, and 10m. of Na 2C03 is
equivalent to 1,000/53 = 18.87 cc. of N Hel. Therefore,
1.6250 Om. will require 1.6250 X 18.87 = 30.66 cc. of N Hel, and
30.66/30.20 = 1.0152 N, the factor of the add. The solution
may be used for all practical work as standardized, the number
of cubic eentimeters used being multiplied by the faetor to reduce
the solution to tmms of exact normality.
If an exactly normal solution is wanted, the dilution required
may be calculated as follows:
30.66 - 30.20 = 0.46 cc. of water to be added to each 30 2 ce
of the standard solution to make it exactly normal. To .make
1,000 cc. of normal solution, it would be necessary to dilute with
water in the ratio, 0.46 :30.66: : x: 1,000 = 15.00 ee. of water in
1,000 cc. of the diluted solution. Measure 15.00 ec. of distilled
 PRINCIPLES OF VOLUMETRIC .ANALYSIS 81

water into a dry 1,000 ce. flask, fill to the mark with 1.0152 N
Hel, mix the solution well, and verify the normality of the solu-
tion by titration against weighed samples of sodium carbonate.
Volumdl'ie solution:;.; of hyclroehlorie acid may be standardized
gravimetrically by determining the amount of ehloride ion
present as silver chloride, as described under gravimetric deter-
mination of chlorides, using 10 ec. portions of the acid as samples.
Ten cubie centimeters of N Hel is equivalent to 1.4334 Gm. of
Agel. If a 10 ee. portion of ttcid ii:i found to yidd 1.4634 Gm. of
Agel, the acid is 1.4634/1.4334 = 1.0210 normal.
Solutions of hydrochloric acid may also be standardized by
titration against standard solutions of KOH or N aOR. Thus if
25.20 ee. of 0.9505 N NaOH is required to l'xactly neutralize
1': 00"
2a. ce. 0 f','d it. ac.t
.Lel, tl'f or o.ftlIe aCIC
"'1,25.20 X 0.9505 -- 09531
IS - . 25.0()-- ."
normal.
The normal Holution is too strong for mnny assay procedures,
so 0.5 N, 0.2 N, 0.1 N, etc., ::.;olutions are employed. These may
be prepared by dilution of the normal solution in the proper
ratio, but the resulting ;;olution should always be l'estalldardized.
Standard solutions of 11ydroehlorie acid pres'erved in tightly
stoppered, alkali:.frec, glass bottles retain their strength in-
definitely. ..
Exercise 11
Preparation and Standardization of Alkali Solutions.-Solu-
tions of sodium hydroxide or potassiulll hydroxide may be
standardized against standard solution of hydrochloric or sul-
furic acid or by titration against an accurately weighed quantity
of pure potassiulll bitartrate or potassium biphthalate.
Object.-To Prepare and Standardize Normal Sodium Hydrox-
ide Solution.
Materials Required.-Aboltt 50 GIll. of sodium hydroxide.
A saturated solution of barium hydroxide.
A standardized solution of hydrochloric acid.
Procedtlre.-l. Weigh about 50 Gm. of sodium hydroxide OJl a rough
balance flnd dissolve it in about 1,000 cc. of distilled watel.'. Add slowly
with stirring a satumted solution of pure barium hydroxide until no further
precipitation occurs (2 to 3 cc.). Allow the precipitate to subside for about
10 hr. and decant the liquid tll'l'ough a filter into a hard glass bottle OJ' a
82 QUANl'I'l'fl'1'IFE PIIARMACEUTICAL CHEMISTRY

bottle the inside of which has previously been coated with paraffin. Close
the bottle tighlly with u rubber stopper provided with a soda-lime tube.
Morc thaH ::t mole, 40.00 Gm., of sodium hydroxide is weighed
out, since it id hygroscopic, and if only a mole were used the result-
ing solution would be below normal strength. The solution is
tre:Lted with harium hydroxide to precipitato any carbonate
fOl'lll8Cl through the aetioll of carbon dioxide on the sodium
hydroxide. The solution is preserv(~cl in a tightly stoppered
bottle fitted wit.h a Hoda-lime tube to prote(:t it from the carbon
dioxide of the air. Solutions which contain carbonate are not
suitable for titration with phenolphthalein as indicator, but when
methyl orange is used the l'csultfl are the same as if all the sodium
were pl'eSfmt combined as hydroxide.
2. Accumtdy measure 30 ce. of llormal hydrochloric or normal ~ulfuric
aeid, diluto with 50 ceo of carbon dioxide-free distilled water, add 2 drops of
phcllolpitt.lmlein '1'.B., and titrate with the Rodium hydroxide solution to the
production of II pUl'IlHLllcnt pink color. If the end point, is passed in the
titmtioll, add :\ fow ellbil~ eentillloters more of acid and again titrate to
the end point. Titmte Cloveml pnrtions of the ::Llkali and from the mean of
the result;; i:aleuhtte the uormality of the sodium hydroxide solution.
A large volume, about 30 ee., of solution is used for each
titration to minimize error from measurements. Every precau-
tion should be taken to insure the coned measurement of the
volume of solutions. The burettes should be eleaned, freed from
air bubbles before titmtion, and read 'with the greatest possible
accuraey.
The soilltion can be made exactly norinal by dilution in the
same manner as that explained under the preparatioll of llormal
hydrochloric acid. Assuming that 30.00 cc. of alkali solution
required 30.15 C:(~. of 1.0119 N HeI, -the faetor of the sodium
hydroxide solution '\vould be
30.15 ~o 1.0119 = 1.0170.

Standard solutions of potassiulll hydroxide can be prepared

in the same way using 1 mole, 56.10 Gm., of KOH per liter. Half-
normal and tenth-normal, etc., solutions can be prepared directly
or by proportionate dilution of stronger solutions, but they
should always be restandardized when prepared by the latter
method,
 PRINCIPLES OF T/OLUMETIUC ANALYSIS 8:3

Sodium hydroxide or pottLSSilllll hydroxide solutions eun abo be

standardized against standard sulful'ie aeid or ugnin;;t :W!'ll-
rately weighed samples of Plll'(" dry potaKsilllll billhtlialate OJ'
potassium bitartrate. The alkali convc-rts the potlliiSiull1 :1!'id
tartrate to potassiulll and sodium t:J.rtrat(~ or Jlota~silUn tartrate
when sodium or potassiulll hydroxide solution, }'('sjJPf:tin\ly,
is used
KHC 4H 4 0r, + NaOH------7I\:NaG H 0 + H~O
1 1 6
188.14 40.00
One thousand cubic centimeters of N NaOH ('oJltain" 40.00 Gill.
of alkali and is equivalent to 188.14 Gill. of potn:"C'iulll hitartrate.
Therefore, 1 cc. of N NaOH iR eqllivalellt to O.IStn Gill. of
potassium bitartrate. From the abovp equivalent the normality
may be calculated.
Exercise 12
Object.-To Prepare and Standardize 0.1 N Solntioll of
Barium Hydroxide.
Materials Required.-18 Gm. bariulll hy(lroxi(le.
0.1 N hydrochloric acid.
Procedure.-DiRsolve about 18 GIll. of P1ll'(' bnrilllll hydroxide in 1,000
cc. of recently boiled, cooled, rli~tilled water and filter the solution quiekly
if turbid. Standardize the solution againRt N Hel llsing plwnolphthalein
as irldicator. It is not !tdvisable to adjuHt, the ~ol\llinn to eX'H'lly 0.1 N,
since it usually becomes t.urbid Oil dilution an(1 rleteriorate;; l'Ulii,lly. Tho
solution should always be standardized before usillll;. Tt Hhonld 1m protee:ted
from carbon dioxide of the air in It well stoj1pcred bottle fitipd with n ~Odll
lime tube.
Barium hydroxide solutions change in concentration rapidly
because the absorption of CO 2 from the l1jr rc:-mlts ill the f orml1tion
of insoluble barium earbol1n,te as follow~;
Ba(OH)2 + CO ------7BaCOa + H 0
2 2

The explanation of the proeeclurc anti tho calenlations if! similar

to that given under the preparation of normal sodium hydroxide
solution in the preceding exercise.
Exercise 13
Object.-To Prepare and Standardize Normal Sulfuric Aeid.
Materials Required.-30 ee. sulfuric acid.
84 QUAN'l'ITATlTTE PHARllL4.CEU'l'ICAL CHEMISTRY

Normal sodium hydroxide solution.

Procedure.-Add 30 cc. of sulfuric add in small portions to 1,020 ce. of
distilled water with constant stirring. Allow the ;,;olution to aequire room
temperature. St,auclardizo till' solution hy titration against normal sodium
hydroxide solution using phenolphthalein ns imlicntor. Calculate the
nOl'mlllity factor ancl writc it Oil the label or adjust, the solution to exact
norlllalit,y.

Since sulfuric acid iN a dibasic acid, 1,000 ee. of [L normal

solution shonld contain 98.08/2 = 49.04 Gm. Sulfuric acid
(95 per cent) has a specifie gravity of about 1.83 at 250. COll-
sequently, 30 ce. of the concentrated aeid should contain about
30 X 1.83 X 0.95 = 52.15 Gm. H 2S0 4 The method of cal-
culating the normality of the solntioll is similar to that described
under the preparation of normal hydrochloric acid (page 80).
The starldardization of sulfuric acid solution call also bc
effected by titration against pure sodium carbonate as des(~ribed
under the standardization of hydrochloric acid. It may also be
standardized gravimetric:ally by precipitation and weighing the
sulfate iOIl as barium sulfate as de~cribed under the grnvimctri<:
determination of sulfate in a soluble sulfate.
Standard Holutions of slllfnric acid preserved in tightly stop-
pered, alkali-free bottles do llot deteriorate.
Questions and Problems
1. Define thn following tl'rms: (a) uorIJlnl solutioll, (IJ) molar solution,
(c) molal solution, (d) gram equivalent weig;ht, (e) normalit.y fttetor.
2. How llluch of eaeh of the following; reugents would be required to
make 500 ce. of 1 N, 0.5 N, 0.1 N, and 0.020 N Rolnti()ll~, respectively?
(a) HOI, (b) H 2S04, (c) NaOH, (d)Na2CO", (e) Ba(OH):.,
3. Cnleulate the llormalit,y of oneh of the following: (a) a solution of
HCI containing 54.7 Gm. per liler, (b) l1 solution of NIl,CO a eontaining 32
Gm. per 500 ce., k) a solution of H 2S0 4 containing 9.8 Om. pCI' 200 ce., (d)
a solution of Ba(OH)~ containing 1.6 Om. per 40 ce.
4. If 1.1200 Om. of pme Na,CO, is ncutmlizcd by 40.2 Qe. of HCI solu-
tion, what is the normality of the HCI?
5. If 20.5 cC. of alkali solution neutralizeH 4.5020 Om. of pure KnC~H100,
what is the normality of the alkali solution.
6. How much 0..1 N yolullletrie solutioll could be prcpflrccl from eaeh of
the following? (a') 100 (~C. of 0.0500 N HCI, (b) :250 ce. of 0.1235 N H~SO<,
(c) 50 ce. of 0.9850 N NaOn.
7. If 40 ce. of HCI solution yielded 0.2140 Gm. of AgCI upon precipita-
tion of the chloride iOllllS silver chloride, what was the normality of the HCI
solution?
 PRINCIPLES OF FOLUME'l'RIC ANALYSIS 85

8. 25 ce. of Hel solution b found to be equivulent to O.S Gm. of pnre

sodium earbOfl[lte. How mueh of c:wh of thc fol1011"ing pnre l'Ntgcllts would
be exactly equivlllent to 25 ceo of the hyclroehlorie (tdll? (a) N:IHCO. 1,
(II) KOH, (c) Ba(OH)" (d) Ca(OHh
9. An unknowll soJut.ion of alkali weighing 10.5420 Gill. required 31).40
ee. of O.OStiO N acid for Ilclltraliz:lt.ion. Caleulatc the J1('n~('ntug() of alkitli
present if the unknown were Na,CO", Bn(OH)2, OT NaOH, l'Psp!)etively.
10. How much 0.1240 N H,SO. would be required to rwntl':1Iizc 25 ec. of
0.0500 N NaOHj 40 ce. of 0.1020 N NaOHj [,.nd 50 ceo of 0.5342 N KOI-l?
11. How lllany cubic centimetcrs of eaeh of the following reagents an~
requirl;lcl to make 500 ce. of 1.0000 N, 0.1000 N, 0.020() N, 0.0100 N solutions,
respectively: (a) 3G per ccnt. HCI, spedne gravity 1.12, (/J) 10 per epnt Hel,
f'pecif1(1 gravity 1.05, (cl 75 per cent 1-1,80" specific grtwity l.{j7, and (d)
10 pel' ceni; H,BO." specifin gravity 1.07?
 CHAPTER V

ALKALIMETRY
Alkali hydroxides and carbonates, etc., are usually titrated
directly with standard solutions of hydrochloric or sulfuric
acid using methyl orange as indicator. Phenolphthalein and
methyl red when used as indicators in such titrations are affected
by the acidic carbon dioxide liberated during titration so that the
end point appears before neutralization is complete. If phenol-
phthalein is used as indicator, carbonates must be removed by
precipitation before titration, or the titration mixture must be
boiled to expel the carbon dioxide formed during titration.
Direct titration is conducted by adding a standard reagent
solution in measured quantity to i substall(~e in solution until the
end point, as shown by a change of indicator color, is reached.
Substances which are insoluble in water or which do not yield
a sharp end point that coincides with the stoichiometric point
upon direct titration are titrated residually.
Residual titration is conducted by treating the substance under
analysis with an amount of standard solution known to be in
excess of that actually required to react with it completely;
the amount of standard solution in excess is then determined by
titration with another standard solution.
Exercise 14
Object.-Assay of Sodium Bicarbonate.
Materials Required.-About 8 Gm. of sodium bicarbonate.
Normal sulfuric acid.
Procedure.-tlDry about 3 Gm. of Sodium Bicarbonate to constant weight
over sulfuric acid, weigh accurately, mix it with 25 cc. of distilled water, and
titrate with normal sulfuric acid, using methyl omuge T.S. as the indicator.
Each cubic centimeter of normal sulfuric acid is equivalent to 0.08400 Gm.
of NaHCO a."

It is a general rule that when the substance to be assayed is a

solid, the accurately weighed quantity to be used should be dis-
86
 ALKALIMETRY 87

solved in sufficient water to' make the solution of about the same
concentration as that of the acid to be used in the titration.
Methyl orange is used as indicator because phenolphthalein and
most other indicators arc affected by the carbonic acid liberated
in the reaction showing a change of color before the reaction is
complete:

2NaHCO a + H SO2 C ---7Na 2S04 + 2H 0 + 2C0

2 2
2(84.00) 98.08

Since sulfuric add haH a hydrogen equivalent of t,yO, 1 cc.

. llormaI so1u t"1011 IS eqmva
of Its . Ient to 2x168
1,600 = 0.0840 Om.
NaHCOa.
Calculate the percentage purity of the sample assayed.
Questions and Problems
1. Write ionically all equations for all reactions that occur in the above
assay.
2. Could normal acetic !leid be used to titrate the bicarbmU1tfl in place of
sulfuric acid?
3. IJook up the pH at which methyl orange exhibits its acid color and
explain why it is a good indicator to use in this titmtioll.
4. If exactly 3.0 Gm. of NaHCO s al'e dissolved in 25 ce. of distilled watef
in the above assay, what would be the normality of the solution? DODS
the normality conform to the general rule stated under Explanation of
Procedure?
6. Enumerate the official carbonates and bicarbonates assayed by similar
methods.

Exercise 15
Object.-Assay of Sodium Hydroxide.
Materials Required.-1.5 Gm. sodium hydroxide.
Normal sulfuric acid.
Procedure.-l. "Dissolve about 1.5 Gm. of Sodium Hydroxide, accu-
rately weighed, in about 40 ce. of recently boiled and cooled distilled water.
Cool the solution to 160. and titrate with normal sulfuric acid, using
phenolphthalein T.S. as the indiclLtor. At the discharge of the pink color
of the indicator, record the volume of acid solution required."

The base, NaOH, behaves in the same manner when either

phenolphthalein 01' methyl orange is used as the indicator.
Consequently, the end point obtained with phenolphthalein as
88 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

the indicator represents complete neutralization of all of the

NaOH as represented by the equation:
2NaOH + H2SOr-+Na2SO'1 + 2H 0 2

2(40.00) 98.08 142.05

In a cold solution, with phenolphthalein indicattll', the end
point of titration of sodium earbonate with Htandal'd H 2SO,i is
exhibited when the solution is one-half neutralized or when the
Na 2 COa is transformed into NfLHCO a.
2Na2COa + I-hSOr--)Na S04 + 2NaHCO a
2

2(105.99) 2(84.00)
2. "Add 3 drops of methyl orange '1'.8., and continue the titration until
the production of a penmmcnt pink colOl'. Ell.ch cubic. centimeter of total
sulfuric acid consumed is equivalent to 0.0400 Gm. of NaOH. Each cubic
ecntililcter difference bet.ween the Humber of cubic: centimetel'H of normal
sulfuric acid COIlsumed in the methyl orangc aIHI phenolphthalein titratiolls
is equivalent to 0.1060 GIll. of Nu 2C0 3 ."
The base NaHCO s is praetieally neutral toward phenol-
phthalein because thc hydrogen ion concentration of the weakly
ionized HCO'3 ion (C0'3 H+ + CO'3) is about the same
(1 X 10- 9) as that necessary to change the color of this intlieator.
A minute amount of H 2S0 4 is sufficient, therefore, to indicate a
neutral solution by the discharge of the pink color. When-methyl
orange is used as indicator, it doc's not change color from alkaline
to acid until a hydrogen ion concentration of about 1 X 10- 4
is obtained. This will occur only when all of the bicn,rbonate
has been noutralizetl. The reaction is indicated by the equation:
2NaHCO a + H2SOC---+Na2S0.j + 2C0 2 + 3H 20
2(84.00)
The method of calculation is as followH: If a 1.0000 Gm.
sample of NaOH required 20.80 cc. of N IhS04 to titrate to tho
end point with phenolphthalein indicator and an additional
0.95 cc. of N H 2S0 4 to titrate to the end point with methyl
orange indicator, the total alkali ealculated as NaOH .would be
20.80 + i.;go~ 0.0400 X 100 = 87 pel' cent. Tho volume of
N H 2S0 4 required to titrate the NaHCO a, after the titration to
an end point with phenolphthalein, represents one-half of the
acid that would be necessary to neutralize the N a 2C0 3 -originally
 ALK"iLIMETRY 89
present. The equivalent weight of Na 2CO a is 105.99/2 X
1000 = 0.053 Gm. pel' cubic centimeter of N H 2SO,j. Conse-
quently, by doubling the equivalent, the result gives the per cent
0.95 X 01>106
of Na 2CO a, e.g" 1.0000 X 100 = 1.01 pCI' eent. The
same result would be obtained by doubling the volume of acid
used and reta.ining the equivalent 0,053. (See eal(~ulation of
K 2CO a ill solution of potassium hydroxide, N.F. VI.)
Questions and Problems
1. What volume of 0.5 N H~SOd will be required to titrate 1.1124 Gm.
of pure Na,CO, using phenolphthalein as indicator in a cold ~()llltion? \Vhnt
volume will be required to titrate the same weight of sample using methyl
orange as indicator?
2. A sample weighing 1,0202 Om. and cOllsisting of a mixture of eqllal
parts of Na,CO, and NaHCO, is iitratrd with 0,82-12 N H 2SO., with phenol-
phthalein in cold solution, and then with methyl orange. What volumes of
the H 2S0 4 solution will be neet'ss:U'y in e:teh titration '/
3. A 25.00 ec. Rumple of KOH :5Qlution rl\<\\\inrl 22,25 ce. of N H2S0, to
titrate to the cud point with phenolphthalein l11dieator in the cold and 4.45
ec, of the same aeid to titrate tn the end point, with methyl orange inclienioL
Calculate the per cent of total uJkali as KOH and the per ceut of K"CQ, in
the solution.
4. Calculate the per cent of Nn,CO.1 !1nd of NaHCO. in a 1.2500 GIll.
sample whieh on titration with 0.2000 N HCI requirecI3(i,25 ee. with phenol-
phthalein indicator and an additional 42,80 co, with methyl orange indicator.
Exercise 16
Object.-Assay of 6jodium Salieyltite.
Materials Required.-2 Gill. of sodium sulicyl!!te.
100 ee. of ether.
0.5 N hydrochloric acid.
Bromphenol blue, T.S,
Procedure.-l. "'TraIlHfer nbollt, 2 Om. of Sodium Salicylflte, previollsly
dried to constant weight at lOODC, :tnd tlceurately weighed, to a tall benker
of about 300-ce. capacity, nnd add 75 ce. of etber IJ.nd 10 clro]Js of brom-
phenol blue T.S. Titrate the mixture with half-mmnnl hydroehlorie add,
mixing intilnlttely the aqueous and ethereall!lyers by vigorous stirring until
;1 IJermanent, plJ.le green color is produced in the ttqucous layer."

The sodium salicylate reacts with the added acid to form freo
salicylic acid and sodium chloride as follows:
C 6H.1.OH.COONa + HCl---7C H 6 4 .OH.COOH -/- NaCI
160.04 138~05
90 QUANTITATIVE PHARMACEUTICAL CHEMIS7'RY

The salicylic acid dissolves in the ether layer almost as rapidly

as it is formed when the ethereal and aqueous layers are well
mixed.

2. "Transfer the contents of tho beaker to a small !!eparatol', and draw off
the aqueous layer in to a small flask. Wash the ethereal layer once with 5 cc.
of distilled water, and add this to the aqueous layer. Add 20 ce. of ether to
the combined aqueous solutions, and mix intimately. Continue the titration
with vigorous shoJcing until a permanent, pale green color is prOduced in the
aqueous byer. Each cubic centimeter of holf-normal hydrochloric acid is
equivalent to 0.08002 Gm. of COH 4.OH.COONa."

The separated ethereal layer is washed with water to remove

small amounts of sodium salicylate which may remain dissolved
therein. Upon washing the combined aqueous solutions and
mixing intimately, any rernaining salicylic acid partitions itself
into the ethereal layer leaving only the Hel in the aqueous
layer to influence the end point.
Questions and Problems
1. Why is bromphenol blue a good indicator to use in the above assay?
2. Compare the above method of assay for sodium salicylate with that for
the assay of tablets of sodium salicylate. Which method is more simple and
rapid? What component of the sodium salicylate molecule is assayed for
in each case? On the basis of the therapeutically active component of the
molecule, which method is preferable?
3. List the official salicylnte and benzoate compounds with the indicator
used for each. Explain why the indicators used are 01' are not suitable.
4. Write equations for the reactions that h1ke place in the assay of:
(a) sodium benzoate, Cb) elixir of sodium salicylate, Cc) ~trontium salicylate.
6. Show how the equivalent is derived for each of the assays in question 4.

RESIDUAL TITRATION METHODS

Residual titration or "back titration" is frequently used when

a reaction proe-eeds slowly or when the substance to be assayed
does not give a distinct, sharp end point with an indicator by
direct titration. Residual titration is carried out by dissolving
the substance under examination in an accurately measured
quantity of standard solution known to be in excess and titrating
the excess of the latter with another standard solution. Numer-
ous examples of this method of titration will be met with in
subsequent exercises.
 ALKALIMETRY 91

010'0
'"
~M~M~OO 00 ~O 00 ~ ~O'l"""OO~OOM"""""""...fl
O'lC')e-l r-I,....IO 0::> 000 QO "Ifl
~~~R~~~8e~~g~88gg~g
~___~~oo=~~~~n=~~Oro
M ....... O!.OQ':l~l:r.ioOO:OoC')
tCr~_It":lf':::OOOOO_-Cl
QOOO~OO~OO~OOO~OOMO 00000000-000
0000000000000000000 000000000000

~OOOOO_OOOOWOWOOOO~ 1t:'I-Cr-IOOO ...... 1"""'I~_,....;IC

O~~~~~O~~~~O~O~~~~O 000""';""';"";000000
92 QUANTl'l'ATll'E PILlRMACEUTICliL CHEMISTRY

Exercise 17

Object.-Assay of Zinc Oxide.

Materials Required.-1.5 of zinc oxide.
N orm:!l sulfuric aeid.
Norml1l sodiulll hydroxide.
Procedure.-"Digest about 1.5 Gill. of freshly ignited Zine Oxide,' accu-
ratdy weighed, with 50 ce. of norlllal sulfuric :wid until solution is com-
plete. Then titrate tIn; eXlil'SS of J,mIfuric neid with llorlllnl sodium hydroxide,
using methyl l'erl-T.S. as indicator. Each l'ubie centimeter of normal
sulfuric !l.cid correspol\ds to O.040(i\) Om. of ZnO."
Thc zine oxide is digested with N H~SO.i to effect complete
solution. The excess acid is then determined by titration with
N NaOH:
ZnO + H SO,c---'ZnSO.j + H 0
2 2
81.38
H~SO.I + 2NaOH---Na SO. + 2H 0 2 2

2 ~\3~oO
, = 0.04069 Gm. ZnO eqnivalent to 1 ce. N H 2SO.j

Calculate tho percentage of zine awl the percentage of zine oxide

in the sampl,e Hssayed.
Questions and Problems
1. Exphlin why zinc oxide is soluble in sulfuric [Icicl, employing the
solubility product principle.
2. CllJcuhtc the l1Jllount of ZllCO" ZnS04, amI ZnCl 2 equivalent to 25 ce.
of 0.0245 N H aS0 4
3. How much 1.0520 N ILSO.j would be required to l'm,Wt quanLitatively
with 1.35GO Om. of ZnO if the ZnO were 90 ucr cent pUl'e?
Exercise 18
Determination of the Purity of the Alkali Salt of an Organic
Acid. The assay of the acetates, citrates, tartrates, etc., of alkali
metals is based upon their conversion by ignition into the corre~
sponding carbonates. All of the alkali combined as carbonate is
determined by titration in the usual way and from the alllount
of carbonate found, the quantity of organic acid salt originally
present may be caleulated. The salt should, as a rule, be dried
previous to ignition in order that caleulations may be made on a
moisture-free basis. . ,
Object.-ARsay of Potassium and Sodium Tartrate.
 jlLJ(ALIMETRY 93
Materials Required.-2 Gm .. of potassium and sodium tartrate.
0.5 N sulfuric acid.
Procedure.-l. "Heat about 2 Gm. of the salt, !U;curately w(ighed, in a
platinulIl or porcelain eru(:iblc, heating; at iirst vpry gPIllly thell "I":ldually
raising the tempemtnl'c until the 8nlt is thoroughly ~arl.lOniz:"cl. (2(lIIiil)n~
Do not use pbtinlllll crucibles for lithium 8tllts.) The fiual temperature
must not exceed it dull red heat and tlw flame of the burner IllUHt not come
in contact with the carbonized ll1ass."

The salt is heated slowly at first, since the sample "",dis and
fuses with concomitant decomposition. If strong heat is applied
in the initial stages of ignition, there may be loss of a })Gl'tion of
the sample through decrepitation or spattering. After the mass
in the crucible is partially charred and white fumes arc no longer
evolved, it is ignited to dull redness. If heated too strongly or
if the flame comes in contact with thc carbonizedl11ass, the alkali
carbonate may be tonvcl'tcd into Lhe oxide.
2. "After [tHawing the e,lrbonized lllass to cool, llisintegl'tltc it with the
aid of a stout glass roeland lnmsfcr the mass and cnwible to [l bC[lker. Alid.
50 ce. of distilled water [Inc] 50 ce. of half-normal sulfuriC) acid, covel' the
beaker with It \V:~tch glass nnd boil the contonts for thirty minutes. 'I'lli'll
filter the Holution and wash the residuo with hot dbtilled ",rlter until the
washings <:oase to reelden blue litmus paper. Now determine the rc~idual
acid ill the cooled filtrate by titratioll with lu~lf-llormal sodium hydroxide,
using methyl orange T.S. as the indicator. The volume of half-normal
sulfuric acid consumed, multiplied by the propel' equivalent of the salt,
represents the quantity of the Balt present in the quantity takon."

The fused residue is broken up by means of a .strong glass rod

over glazed, black paper and transferred with thc crucihle into
a 250 cc. beaker. About 50 cc. of distilled water and 50 ee. of
sulfuric acid are added along the glass rod, and the mao;s is com-
pletely dissolved.
The reaction taking place upon ignition \vith free advent of
oxygen from the air may be represented [lS follows:
KNaC 4H 4 0 6-KNaCO a + 3C0 2 + 2H 0 2

210.12
and upon the addition of an excess of acid the double carbonate
is converted into double sulfate:
KNaC0 3 + H2S0c~KNaS04 + CO + H 0 2 2

122.1 98.08
94 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The volume of acid in excess of that ~leccssary to react with the

carbonate residue is determined by titratioll with stn,ndal'cl alkali
solutioll. Thus, if 12.20 ec. of 0.5 N N aOH is requil'f~d to neutral-
ize the exceSH acid, 37.80 ceo of the 0.5 N H 2S0 4 must have reacted
with the (~arbon(1te in the residue.
One mole of the tartrate upon ignition forms one mole of the
carbonate whieh is equivalent to one mole of H 2SO.j . Sulfurie
aciel haH a hydrogen equivalent of two, and thn ~tall(lard solution
m,;ed is 0.5 N. Each cubic centimetcr of 0.5 N acid consumed in
the titration is, therefore, equiyalcnt to:
210.1.2
2 X 2X UlOa -
_ 0 0l':2r':'l G
. LI L1.)
ITN ft C'elI::I"j_10 6
m. \.

The U.S.P. requires that potaRsinm anel sodium tartrate con-

tain not less than 9H per cent of KN[LC1H 1 0 0 Calculate the
purity of the sample annJyzell and compare the result with the
U.SeP. requirement.
Questions and Problems
1. Why cannot t.ll[~ alkali saH~ of sueh acids as citric, tartlll'il', ('te., be
tit.rated directly with II strong acid, in the sallle manner aH HolnLle carbollates
Ill') titrated?

2. How much of each of the following snits is equivn.lollt- to 1 (C. of 0.5

N H,SO'l? Ca) KHC1H.10" (IJ) K 3 C,rLOdLO, (c) CH;,COONlt, (Il)
Li 3CGH,O,.4H 20.
3. Name n number of oflicia! sllbstullce:, :I.~mycd by 1he above method.

Exercise 19
Object.-Assay of Magn(~si:1 MagmfL.

Materials Required.-5 Gm. of l1lagncHia magma.

N orlllal sulfuric acid.
N onnal sodium hydroxide.
Procedure.-" AUl'!' thorollgh (1git.atioIl, place ahout 5 Om. of Magnesia
Mn.gma in a tared flll~k, stopper, :mri weigh accnralply, arid 25 N~. of UOrlllf1!
sulfuric aeid, aIlLI nfter solution is cOlllpletc, titn,tfl t.h(~ l'Xl.l'SS of ncid with
normal sodium hydl'oxidn, llsing mothyl red '1'.8. as tho indicator. Eaeh
cubie centimeter of nornm! ~ulfurle acid iR oqnivalent to 0.02017 Om. of
Mg(OHh"

The magnesia magma is cliKsolvecl in an accurately measured

excess of normal sulfuric acid solution to insure eomplete neu-
tralization of all of the magnesium hydroxide with the formation
 AL[{ALIIYfETRY 95
of the soluble lllagnesium slilfate. The excess acid is then deter-
mined by l'ei3iclual titration with normal sodium hydroxide, using
methyl orange as inclieator.
The reactiolls ,yhich take place when the magnesia magma is
dissolved and upon reflichml titration are a~ follows:
Mg(OH)2 +
H 2SO,r--->l\:IgSOj 2Hi) +
58.34
H 2 S0 4 +
2NaOH-----.;Na 2SO,j + 2H~O
Each cubic centimeter of N lU30 4 cOllHumccl by the magma ii3
equivalent to:

2~\~~06 = 0.02917 Gm. Mg(OH)2

If a sample of magnesia magma weighing 5.2430 Om. when

dissolved in 25 ce. of 0.9m5 N H 2SO'1 required 9.85 cc. of 1.1402
N NuOH to titrate tllH excess acid, the per eent Mg(OH)2 in
the sample could be c,doubted from the following formula:
[(1'1'. H 2SO,j X N) - (cc. NaOH X N)] X equiva]Pllt
Wt. of sampIe~
X 100 = pel' cont, 01' suhstituting the figures from the example
eitecl aboye,
[(25 X 0.9915) - (9.85X 1.1402)] X 0.02917 X 100 ==
5.2430
7.54 per cent.
The U.S.P. requires that magnesia magma contain not lesi3 than
7 and not more than 8 ..5 per cent Mg(OH)z. The above sample,
therefore, conforms to the official requirements.
Questions and Problems
1. Why is l'(~siuu[ll titratioll resoyted to in this assay?
2. Write all n~aetiOJlR iouieally.
3. To how Illul'h Mg(OH), i;; 1 ee. of 0.1054 N HCI equivalent?
4. 12.32 GIll. of maglll!1 magnesia dissolved in 50 cc, of 1.0340 N H 2S0 4
required 24.00 ce. of 1.1255 N NuOH for rcsidmtl titrntion. Ca!eulate the
percentage Mg(OH)2 present in the sample. To how much MgO does the
result corl'c,~pond?
Exercise 20
Object.-Assay of Methenamine.
96 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Materials Required.-l Gm. of methenamine.

N oJ'mal sulfuric acid.
Normal sodium hydroxide.
Procedure.-l. "Place about 1 Gn~. of Methenamine, accurately weighed,
in it beaker, add 10 cc. of normal sulfuric acid, alld evaporate on a water
bath (or boil gently adding n little di~tilled water from time to time, if
necessary) until the Ullor of forlllaidehyde is no longer perceptible."

The weighed sample of methenamine is heated on a water

bath with a measured excess of normal sulfuri(~ acid to decom-
pose it into ammonia and formaldehyde, distilled 'water being
added from time to tillle to replace the water lost by evaporation.
Completion of the decomposition is indicated when the odor of
formaldehyde is no longer perceptible. The decomposition
reaction may be indicated in t,vo steps aR follows:
(CH2)oRl + 10H20-~6CH20 + 4NH 40H
140.18
4NH'IOH + 2H 2 SO r --72(NH.1)2S0.j + 4H zO
2. "Cool, add 20 ceo of distilled water, and tiLrate the excess of acid with
norl11l)l sodium hydroxide, using methyl red T.S. as indicator. Each
cubic centimetel' of normal sulfuric acid corresponds to 0.03503 Gm. of
(CH 2)GN . "

The excess sulfuric acid found by residual titration with N

N aOH subtracted from the amount originally added gives the
amount of acid which combined with the liberated ammonia.
Since each mole of methenamine requires 2 moles of sulfuric
acid in the reaction, the quantity of methenamine equivalent to
1 cC. N H 2S0 4 is
140.13 _ 0031':03 G
2 X 2 X 1,000 - . 0 m.

Calculate the percentage purity of the sample assayed.

Questions and Problems
1. Why should not the solution (If ~mmonium sulfate be evaporated to
dryness to volatilize all of the formaldehyde in this assay?
2. Why is sulfuric acid a better acid to use than hydrochloric acid when
high temperatures are involved in the ass!ly process?
8. If 1.12 Gm. of methenamine treated with 80 ce. of 0.5250 N H 2S04
require 12.40 cc. of 0.9885 N NaOH to titrate the residual acid, how pure is
the methenamine?
 ALKALIMETRY 97

Exercise 21
Object.-Assay of Solution of Ammonium Apetutc.
Materials Required.-25 ce. of solution of ammoniulll l1ectate.
Sodium hydroxide, T.8.
Normal sulfuric acid.
Normal sodium hydroxide.
Procedure.--"TnlIlRfer 25 ec. of Solution of AmmoniuJIl A('(Jt:ltc to a
diHtilling ~fllik, dilute with 75 ce. of di~till('[l Wlltf1l", add 50 ne. ot ~()di\llll
hydroxide '1'.8., and distil the liquid until all of tl](~ nmlllollia IlflS been
driven ovor (about 100 ec. of distillate), rcepiYing the distillate under the
slp'face of 50 ee. of normal Rulfuric l1~id cOlltninNI in a flade Titrate HI!'
exeoss of acid with normal sodium hydroxide, using lllethyl red 1'.8. n~
the imlinutor. E!leh cubic eentimcter of normal Hulfuric [wid is eqniY1l1l'nt
to 0.0770(l Gm. of CH:J.COO.NH 1 ." .

The NaOH reacts with the amlllonium acetate liherating NH~

and combining ,,,ith the acetic and earbonic l.1cicls to form non-
volatile compounds. The distilling fbHk should he eonnected
to the condenser at once after adding the NaOH to prevent loss
.
of the volatile NH a. The NHa formed in the re::wtioll,
CH aCOONH 4 + NaOH---7CH aCOONa + NI-L + H~O,
77.06
distils with water when the flask is heated. TI18 NHa is con-
ducted into a receiving flask which contains an excess of N
H 2S0 1 The eondenser should be fitted tightly with an adapter
in sueh a manner that the diHtillate will be collected beneath the
surface of the standard ariel. This is done to prevent loss of
NHa from the distillate. As soon as the NHa in the distillate
comes into the reeeiving flask, it eombines with the H 2 S0 4 to
form (NH.1)SO'I,
2NH3 + H2S04-~(NH4hS()t
17.03 132.14
,>

The exeess acid is then determined by titration wiih standard

alkali.
Questions and Problems
1. Why is methyl red rather tlwn phenolphthalein lweI! in this assny?
2. Write equations for the reaetions that o('em in the aaSlLY of: (n)
aromatic spirit of ammonia, (b) [tmmoniunt bromide in tablets of three
brornidcs, (cl acid ammonium valemte, (d) ammonium earbonute.
 98 QUANTITATIVE PIlARJ.lfACEUTICAL CHEMISTRY

3. Is this method of assay goncrrrlly applicablc 10 ammonium com-

pounds?

Estimation of Nitrogen by the Gunning-Kjeldahl Method.-

Severnl meth~)(ls have been developed for the so-called moist
combustion of nitrogen. The Gunuiug-Kjeldahl method which
will be described lwre, because it is the method speeified in the
National Formulary for the estimation of the nitrogen content
of beef extract, illustrates the general principles imrolvfcl in the
determination of nitrogen by all modifications of the Kjeldahl
method. See also peptone and casein, reagent, U.S.P.
The U.s.P. gives the following directions for the determination
of total nitrogen by the E:_jeldahlmethod:
"Nitrates and Nitrites Absent.-Place about 10m. of the sub-
stance, accurately weighed, ill a 500-cc. Kjeldahl flask of hard
glass. The material to be tested, if solid or Hmni-solicl, may be
wrapped in a sheet of nitrogen-free filter paper for cOll\'eniellC:p
in transferring it to the flask. Add 10 Gm of powdered potas-
sium sulfate or imhydrous sodium sulfate, 0.5 Om. of powdered
cupric: sulfate, and 20 ce. of Bulfuric: acid. Incline the flask at an
angle of about 45 and gently heat the mixture, keeping the
temperature below the boiling point of the mixture until frothing
has ceased. Increase the heat until the acid boils briskly, and
continue the heating until the solution has been clear green in'
color for thirty minutes. Allow the mixture to cool, add 150 cc.
of distilled water, thoroughly mix the contents of the llILHk, and
cool again. Add calltioutlly 100 ee. of a 30 per cent aqueous
solution of sodium hydroxide added so as to eause the ::;ollltion to
fiow down the inner side of the flask to forlll 11 layer under tho
acid solution. Add a. few pi(~eeH of gmnubted zine, connect the
flask, by mea,llS of a Kjeldahl connecting bulb, with a condenser,
the delivery tube from which dips beneath the surface of a, mixtul'u
of 30 ce. of half-normal hydrochloric or sulfuric acid a,n.d 25 cc.
of distilled water contained in all Erlellmeyer flask or n wide-
mouth bottle of about 500-cc. capacity. Mix the contents of the
l(jeldahl flask by gentle rota,tiol1, and diHtillwtil about two-thirds
of the contents of the flask has distilled over. Add about 5 drops
. of methyl red '1'.B. to the contentf; of the receiving flask and deter-
mine the excess of acid by titration with half-normal sodiulll
hydroxide. Run a blank test and make neceHsary correction,"!.
 ...lLf(ALIME1'RY 99
Each ee. of half-normal add consumed is equivalent to 0.007004
GIll. of nitrogen.
"'iVhen the nitrogen content of the substanc(> is knuwn to be
low, the half-normal hydrochloric or sulflll'ie acid lllLly be t'C!Jllueed
by teuth-normal acicl uml tenth-normal ttlka1i should then be used
in t.itrating the eX!)C'HS of aeid. One ('ubic centimeter of tenth-
normal hydrochlOl'ic or sulfuric acid is equivalent to 0.0014008
Gm. of nitrogen.
"With Nitrates Present.-Place a quantity of the fmbstance,
accurately weighed, corrciiponcling to about 0.15 Gm. of nitrogen,

FIG. 14.-A singlo-unit Kjddahl apparatus.

in a 500-cc. Kjeldahl flask of hard gla!'s, and add thereto 2,5 cc.
of sulfuric acid in which 1 Gm. of salicylic Heid has previously been
dissohcd. Mix the c:ont(~lltR of the flask thoroughly, and allow
the mixturp, to st.and for thirty minute::; ,dth frequent shaking.
Add to the mixtlU'G 5 Gm. of powdered sodium thiosulfate and
again mix thoroughly, then add 0.5 GIll. of powdered cupric: sul-
fate and proceed as direc:ted previously for Nitrates and Nitrites
Absent, beginning with 'Inc:lillo the flask at all angle of about 45.'
"N OTE.-There are {:ert!:dn allmJoids and other nitrogen-
containing organic compounds that will not yield all of their
nitrogen to digestion with sulfuric aeid, and this method, there-
fore, cannot be used for the determination of 'nitrogen in all
organic compounds."
100 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The organic matter is decomposed by sulfuric acid with the

aid of a catalytic agent in a Kjeldahl apparatus (Fig. 14). The
carbon and hydrogen are converted into carbon dioxide and
water, respectively. The nitrogen is converted into an ammo-
nium salt. The ammonia is liberated from this salt by a fixed
alkali and distilled into a measured qnantity of arid of known
norll1fl,lity. The residual [lcid is then determined by titration
with a stn,ndard alkali solution, alld the percentage uf llitl'ogPfl
in the sample is calculated from the data obtained. This
method i::; applicable to the determination uf nitrogen in most
organic and ammoni::wal compounds but not to t.he estinuttion
of nitrogen in nitrates unless modifiC'd. It eannot be used for
the analysis of nitl'OSO-, nit1'o-, azo-, and azoxy-compouncls or
for certain other compounds such as pyridine and quinoline and
certain alkaloids, because these cOlllpouncb do not yield all t,lwir
nitrogen when digested with sulfuric acid.

Exercise 22
Object.-To Determine the Amount. of Nitrogen in the
Alcohol-soluble Solids of Extract of Beef.
Materials Required.-A Kjeldahl apparatus.
10 Gm. of extract of beef.
100 cc. of alcohol.
25 cc. of 0.1 N sulfuric acid.
25 ec. of 0.1 N sodium hydroxide.
20 CC. of concentl'llted sulfuric acid.
10 Gm. of potas~ium sulfate.
A 30 per cent solution of sodium hydroxide.
Cochineal 01' methy Ired indicator solution .

. All of the chemieulH llsed must be of reagent quality and be

free from nitrates and ammonium ~mHfl.
Procedure.-l. Wdgh about 10 Gm. of extl'l1ct of beef aeeurately and
add sufficient distilled water to mnke 100 ce.

The extract of beef call be weigh cd cOIlveniently in a small

evaporating dish. It may then be washed into [I, 100 ce. volu-
metric flask ,vith distilled water.
2. Distribute 10 ce. of the solution ovor about 25 Gm. of previously
weighed sand or 5 Gm. of asbestos fiber contained in a previously weighed
flat-bottomed porcelain dish and dried to constant. weight in an oycn at a
 ALKALI M filTRY 101
temperature of 105"0. The weight of the r{)~idue frolll 10 (:e. of the solution
corresponding to 1 Gm. of cxtnwt of beef should not bl:: Ips:'> tklll 0.7.5 Gill.
corresponding to 75 per cent of total sui ids in the original f'xtrnl't.

The pasty mass of the extract iH dilntE'd and clistribnted onr

sand or asbestos to expose it large surface area to the heat and
thereby hasten the evaporation of water. Extruet of beef
containing less than 75 per cent of total solids docs not meet the
National Formulary requirement.
3. Transfer another 25 ce. portion of thl\ solution of beef extraet to an
Erlenmeyer flask, add 50 cc. of o.1cohol, llnd Hhake the mixture vigol'ollsly.
When the preeipitate which forms has suhHided, eollect it on a counter-
poised filter paper, wash it, three times with 5 ce. portio;1S of n mixture
of 2 volumes of alcohol and 1 volume of distilled water, and then dry it to
constnnt weight at 105"0. Reserve the filtrate and wnshings for the
determination of nitrogen.

The precipitate thrown down by the alcohol is com;iderecl to

be the alcohol-insoluble solids of beef extract.
4. Measure an aliquot portion of the alcoholic filtmte fro111 the pre-
ceding determination correspollding to 1 Gm. of alcohol-soluble solids into
a 500 cc. Kjcldahl flask.

The aliquot portion corresponding to 1 Gm. of alcohol-soluble

solids of beef extraet is ealcubted as follows: If the total solids
obtained from 10 cc. of the solution containing 1 Gm. of the
extract of beef weigh 0,8 Gm., the percentage of total solids
in the extract will be 80 per cent. If the alcohol-insoluble solids
from 25 cc. of the solution contftining 2.5 Gm. of beef extract
weigh 0.2 Gm., the percentage of alcohol-insoluble solids in the
total solids will be 2.5 0: .0 X 100 = 10 per cent. Each gram of
8
the extract must therefore correspond to 0.8 Gm. of total solids
and to 0.8 - 0.08 = 0.72 Gm. of alcohol-soluble solids. ThE'
volume of the solution of beef extract corresponding to 1 Gm.
of alcohol-soluble solids ilNhis case will he O.~2 X 10 = 13.9 cc.,
where 10 is the dilution factor.
5. Add about 10 Gm. of powdered potassium sulfate and 20 ee. of con-
centrated sulfuric acid to the solution contained in the :flask. Place the
flask in a slightly inclined position on an asbestos support with n circular
102 QUANTITATIVE PHARMACEUTICAL CHEMIBTRY

opening so that the free flame will comc in direct contact with the bottom
of the flask. HC:lt below the boiling point for about 10 min. or until frothing
has ceased and then mise the temperature and boil the mixture until the
latter acquires a palc straw [Jolor 01' is nearly colurless.
The proteins and other organic matter arc oxidized by the
sulfuric acid-potassium sulfate mixture, leaving the nitrogen
combined as ammonium sulfate. If foaming; OCcurs at first,
it can be prevented, for the most part, by dropping a small piece
of paraffin into the flask. The time nocessary for the oxidation
of the compounds depends upon their nature, from }~ to 12 hr.
heating being required by different compounds and mixtures.
A small erystal of copper sulfate or a globule of mercury may be
added to the digestion mixture to catalyze the oxidation.
6. Cool thc flask, add about 250 cc. of distilled water, [md cltutiously
add a 30 pel' eent solution of sodium hydroxide until the contents of the
flask urc distinctly alkaline. Use phenolph1.lmlein added to the mixture
as indic~ltor. Connect the flask at once to the l(jelduhl tmp, condenser,
lmd receiver as illustrated in Fig. 14, so that the 1011'e1' end of the condenser
dips beneath the Hurface of 25 ce. of 0.1 N sulfurie acid contained in the
receiving flask. Distil the mixture \llltll about 100 ee. of distillate is
obtained.
In making the mixture alkaline, it should be borne in mind
that the pink, alkaline color of the phenolphthalein is destroyed
by a large excess of alkali. A few pieces of granular zinc or
porous plate placed in the flask prior to distillation will prevent
bumping. The alkali liberates the ammonia from the amllloniulll
sulfate, and upon diHti11atioll the ammonia gas is driven over and
collected in the standard acid. In the analysis of compounds
containing a high percentage of nitrogen, all of the standard acid
may be neutralized before the whole of the ammonia has distilled.
If from 2 to 3 drops of methyl red or cochineal indieator are
added to the standard acid before the distillation is started, the
color of the indicator 'will show whether all the acid has been
neutralized, and in case it has, more standard [wid can be placed
in the rceciver without an appreciable loss of ammonia. The
Kjeldahl trap in the apparatus prevents liquid from the distilla~
tion flask from being carried over mechanically.
7. Add methyl red or cochineal indicator solution to the distillate and
titrate the cxcess acid with 0.1 N sodium hydroxide solution. Calculate
t.he percentage of nitrogen in the alcohol-suluble solids.
 ALKALIlIfE'l'RY 103

Deduct th(~ number of cubic centimetrl' of 0.1 N alkali required

to neutralize the exccss acid from the num]wr of cubic centi-
meters of 0.1 N aeiclllsed. The difference will be the number of
TABLE IX.-Ol'FICIAL ALKALIMETIUC ASSAYS.BY REf:'IllUAL TrrnATION

Am(JUnt
w':icd, Indi- Nnr- Equivll-
SUbt:itulH.:e mnlity If!nt of Offif'ial l'enUin~nWtlt8,
Gm.o1' e~'\,tul' per l'!~llt
ce, of achl 1 ej~.

------- --- - - - --- - - '- -----_._----

U.S.P.
Ammonia, aroma-
tic spirit. of (iol'
tot,,1 NH,). . . .. 10 lVI,It. 0.,; D.OOSiili; NH, = 1.7 to 2.1
Ammonium :lC(~-
tate, Rolution of 2G M,R, 1.0 0.07701] CH,COO.NH,=[),5to7.5W/V
Anunoniuln ('ar-
bon"te .... ' .. . 2 111.0. 1. 0 0.0170:1 N1I3 = 30-32
Effervescent pow-
den", compound 2 Pp, 0,5 0.04201 N"HCO,=2:.lto'.!.7
G1ycel'yl trini-
trute. tablets I)f. (LOo' M.R. 0.02 D.OOL;14 ChH:.(NO,1)' = 87.;; to 112.,,'
M:lgnesia magma. 5 lI1.R. 1.0 0.021117 l\Ig(OH), = i tu S.;)
~IagnesiulIl car-
bonate .... , ... M,O. 1.0 0.020lG MgO = 3[}.2 to 41.5
l\Ingnesiuln oxide. 0,5 M.O. 1.0 0.U20lfi MgO = OG
1Vlagncsium oxide,
beavy .. ,,,.... O.r. M.O. 1.0 0.02011l ::\lg0 = IlG
Methenamille, , . . I lI1.R. ],0 0.0:330:1 (Cn,),N, = 00
Potassium n('etaie 2 A1.0. 0.5 O,[J4JIOli CHaCOOK = HI'
Potassium a.nd so-
(liurn tu.rh.ate. 2 M.a. n,ii O. 05~i:;'1 I{NuC>,H,Or. = un
Pota8~iuIll eitratc. M.a. n.5 O.[JiijIJlj J("C"U,07,H,O = UO
Sodium :teetate.. . 2 ALa. 0.,; n. 04!o~ CH,COONu ~ 90
Sodium citrute. . . 1.5 A1.0. O.fJ I),04aOl Na,C,fhO, = D(I
Zinc oxide ..... , , 1.5 l\LH.. 1.0 O.04(Hl(l ZnO = In
Zirw Rtenrntf~, . 1 M,R. 0.1 O,I)OlOlifI ZnO = 13 - 15.5
N.F.
Anunoni um bro-
Inide in tftbh~ts
of threo bl'O-
luides .. "..... O.GI' ;\Ul. O.J1 O.04Sf18 NI'{,Br = :10.8 to 35.8'
Ammonium valer-
ate, acid .. " .. . 1.5 :\['R. n.r. C.,Ir"COONH, = 62 to li7
Calamine, pre-
pared, ....... , 1.5 M,O. 1.0 O. a'iOan ZnO = 08
Iron and anlnlon-
fum acetate,
so1ution of. . . .. 25 lI1.R. O.ii [J.nOS.31ii NH, = 0.0 ttl O,8W/V
IchthulllJIlol (for
NH;~).
, ...... ,' 5 M.lt. 0.5 0.OOS;'[3 NIl, = 2.,;
Lithium earbon-
"tp ....... " ... 1.5 M.O. 1.0 n IWil'4 Li~(,O:! = IlH
Litbium eitn\te.. .
Methenamine,
2 1[,0. 0.5 O.U:l-1llS I C"I{.,.OH.(COOLi)" = US.5
fLmpuh~ (If ..... . l\!'H. 1.0 O.n3iiO:j (CH,loN, = \IJ tu Ill:;'
MethellR.Illine,
tablets of, . . . . . III ::\LH. 1.0 0.03503 (CH,),II., "" U4'to lOG'
Methenamine HHfl
Bmlium bipho8-
pbate, tablets
of. , ... , , . . . . . . 0,25" M,R. O./i 0,01752 (CH,)oN. = 92.5 to 107.5
Zinc oxide in mild "
resorcinol paste M.a. 1.0 O.1)4()09 ZnO = 24 tn 26
Zinc oxide in
strong re-sor-
cino! paste ..... I M.O. 1. 0 0.040(1) I ZnO
= IU to 21
I I
M,O. = methyl ontnge, M.R. = methyl red. Pp. = phenolphthalein.
'. Weight of ingredient sought.
b P~r cent of labeled umoullt.
104 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

cubic centimeters of the 0.1 N acid neutralized by the ammonia.

Since each cubic centimeter of 0.1 N acid neutralized by the
ammonia is equivalent to 0.001401 Om. of nitrogen, the amount
of nitrogen obtained can be found by simple multiplication of the
number of cubic centimeters of 0.1 N acid neutralized by this
factor.
Questions and Problems
1. Name several nitrogen-containing products used in pharmacy to which
the Gunning-Kjeldahl method of analysis might be applied.
2. A nitrogenous substance weighing 6.150 Gm. furnished enough
ammonia, when analyzed by the Gunning-Ejoldahl method, to react with
40.65 cc. of 0.1 N H~S04. What percentage of nitrogen did the substance
contain?
3. The nitrogen in a 5.0 Gm. sarnplc of a food product was determined
by the Gunning-ICjeldahl method. After passing the evolved ammonia
into 90 cc. of 0.5 N acid, 25.80 ceo of 0.1 N alkali was required to titrate the
excess acid. Calculate the perccntage of nitrogen in the sample.
4. How does the pl'Occdul'c for the determination of total nitrogen diffe!'
from that given nbove when. nitrates are present?
 CHAPTER VI

ACIDIMETRY
Acids are estimated quantitatively by methods analogous to
those employed in alkalimetry, namely, by directly titrating an
exact quantity of the acid or acid Sltlt with sta.ndard alkali
solution or by adding an excess of the latter and determining the
amount in excess by residual titration with standard acid solu-
tion. Direct titration is employed whenever practicable, since
it is easier and requires fewer burette readings.
In assaying acids, the quantity of acid to be taken should be
such that about 30 to 40 cc. of the alkali solution will be con-
sumed. As a general principle, it is recommended that the
normality of the acid to be titrated should be approximately
the same as that of the titrating medium. Except when other-
wise directed, the liquid to be titrated should be brought to room
temperature before titration, as many indicators give different
values at different temperatures. Ifor most of the inorganic
acids, methyl orange, methyl red, phenolphthalein, or litmus
can be used as indicators, but the alkali must be standardized
with the particular indicator used. For organic acids, phenol-
phthalein is always used.
DIRECT TITRATION METHODS
Exercise 23
Object.-Assay of Hydrochloric Acid.
Materials Required.---'-3 ce. of hydrochloric acid.
Normal sodium hydroxide.
Procedure.-"Weigh accurately about 3 ce. of lIydrochlol'ie Add in a
tared, glass-stoPPf'red flask. Dilute with about 25 cc. of diAtilled water
and titrate with normal soditun hydroxide, using methyl red T.S. as the
indicator. Each cuhic centimeter of normal sodium hydroxide is equivalent,
to 0.03647 Om. of Hel."
Concentrated acid solutions are weighed in glass-stoppered
flasks to prevent loss of dissolved gases in SOlUe cases p,nd to
105
106 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

prevent the absorption of water in other cases. Samples are

taken by weight rather than by volume, because it is difficult to
measure small volumes of concentrated acids accurately, and a
small error in the measurement of the sample would cause a
large per cent errol' in the result.
Hel + NaOH~NaCl + H 0 2

36.47 40.00
The percentage of absolute HCl in the sample may be calcu-
latd:
cc. N NaOH X equivalent of HCl X 100 t
--W~of sample of He} = per cen,
The U.S.P. defines hydrochloric acid as an aqueous solution
containing not less than 35 and not more than 37 pel' cent of
Hel. Compare the strength of the acid assayed with the
official requirement.
Questions and Problems
1. A sample of HCI weighing 3.0024 Gm. required 34.45 co. of 1.1542 N
NaOH upon til.ration. What per cent of HCI did the sample contain?
What is the Na,CO a titer of the acid? What is the normality of the acid?
What is the molarity of the acid?
2. Calculate the per cent of chlorine in the sample in problem 1.
3. One cubic centimeter of a solution of HCI was found to be equivalent
to 1.12 ce. of NaOH solution. When the HCI was subsequently standard-
ized, its HCI titer was found to be 0.003642. What is the normality of the
NaOH solution?
4. A solution of HCI has a sodium carbonate titer of 0.0530. What weight
of sodium carbonaLe must be taken as a sample in order that the volume of
acid in cubic centimeters may give the per cent of Na 2C0 3 ?
Exercise 24
Object.-Assay of Diluted Sulfuric Acid.
Materials Required.-l0 cc. of diluted sulfuric acid.
Normal sodium hydroxide.
Procedure.-" Accurately measure 10 cc. of Diluted Sulfuric Acid and
dilute with about 20 cc. of di~tillcd water. Titrate the solution with normal
sodium hydroxide, using methyl red T.S. as the indicator. Each cubic
centimetel' of normal sodium hydroxide is equivalent to 0.04904 Gm. of
H 2S0 4."
The official diluted acids are assayed, as a rule, by measuring
an exact quantity of the acid, titrating it after dilution, and
 ACIDIMETRY 107
calculating the per cent of acid on a weight to volume basis.
This method of sampling is more convcnie~t and rapid than
the taking of weighed Ramples, and, Rinee the volume of diluted
acid used as sample is relatively large, the error in measnring is
slight. The sulfuric acid reacts with the sodium hydroxide tL::;
indicated by the following equation:
H 2S0 4 + 2NaOH~Na2S0'1 + 2H 0 2
98.08 2(40.00)
Since sulfuric acid is a dibasic acid and each mole is equivalent
to 2 moles of the monoacidic base, N aOH, the H 2S0 4 equivalent
of 1 cc. of 1 N NaOH will be 98.08/2 X 1,000 = 0.04904. Calcu-
late the per cent of H 2S0 4 in the sample assayed.
Questions and Problems
1. What indicator solutions are suitt>ble for use in the assay of diluted
sulfuric acid?
2. List a number of other official diluted acids, which are nssayed by !1
similar procedure, witli the indicators used in cach.
3. If the sample of H 2SO. assayed in this exercise had a specific gmvity of
1.0640, what would be the per cent by weight?
4. Calculate the sodium carbonate [lnd ammonium hydroxide titers of the
acid assayed in this exercise.

Exercise 26
Object.-Assay of Boric Acid.
Materials Required.-2.5 GIll. of boric acid.
100 cc. of glycerin.
Normal sodium hydroxide.
Procedure ...,-" Dry about 2 Gm. of Boric Acid to constant weight over
sulfuric acid, weigh accurately, and dissolve the dried Acid in 100 ce. of a
mixture of equal volumes of glycerin I1nd distilled water, prcviouHly ncutral-
ized to phenolphthalein T.S. Titrate with normal sodillm hydroxide, using
phenolphthalcin 'r.s. as the indicator. Discharge the pink color by the
addition of 50 cc. of glycerin, neutralized to phenolphthalein T.S., and
again titrate until the pink color reappears. Each cubic centimeter of
normal sodium hydroxide is equivalent to 0.06184 GIll. of HaBOa."

If an aqueous solution of boric acid is titrated with standard

alkali solution using phenolphthalein as indicator, the end point
as shown by the development of the pink color appears before
all of the boric acid has been neutralized. If sufficient glycerin
is added to the boric acid previous to the titration, however,
108 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

the end point is sharply defined, the boric acid behaving as a

monobasic acid. The action of glycerin may be explained by
the assumption that glyceryl borate (C 3H a020H)B(OH)2, a
stronger acid than boric ncid, is formed, thllS preventing the
formation of sodium rnetaborate. The hydrolysis of the latter,
if present, would increase the [OH-] causing the end point of
titration to be reached before an equivalent quantity of NaOH
has been added. Glycerin is frequently acid in reaction and
should, therefore, be neutralized before addition to the sample
being analyzed:
HaBOa + NaOH----->NaH BOa + H 0
2 2

61.84
Therefore, 1 ce. N NaOH = ~~o~~ = 0.06184 Gm. boric acid.

Questions and Problems

1. Why is glycerin added in this assay?
2. Write the reaction structurally.
B. C[tlculate the per cent HaBOa and the per cent B in the sample assayed.

Exercise 26
Object.-Assay of Tablets of Sodium Salicylate.
Materials Required.-20 tablets of sodium salicylate.
Diluted hydrochloric acid.
About 100 cc. of et.her.
2 cc. of neutral alcohol.
Ferric chloride, .T.S.
0.1 N sodium hydroxide.
Procedure.-l. "Weigh not less than 20 of the Tablets, l'educe them to a
fine powder without an appreciable loss, and transfer an !lliquot portion,
cquiv!llent to about 0.3 Gm. of sodium salicylate, to a separatory funnel.
Add 25 ce. of distilled water and a slight excess of diluted hydrochloric !lcid,
and completely extract the liberated s!llicylic acid with ether. Trnnsfer
the ethereal solution to a suitable flask and distil off most of the ether, being
careful not to volatilize the salicylic acid, and allowing the last few cubic
centimeters to evaporate spontaneously."

The tablets are weighed before powdering so that the average

weight per tablet can be computed. When HOI is added, salicylic
acid is set free as indicated by the following equation:
 ACID! ME'l'RY 109

Shake the aqueous liquid successively with 20 ce. portions

of ether. The salicylic acid, being more soluble in ether than in
water, partitions itself into the ether layer. Completion of the
extraction can be determined readily hy withdrawing 1 ceo of
the aqueous liquid and adding 3 drops of ferric chloride, T.S.
If no violet color forms in the test liquid, the extraction lllay be
considered to be complete. If a violet color appears, continue
to extract the aqueoul5 liquid with 20 cc. portions of ether until
a negative test is obtained. Upon evaporation of the combined
ethereal solutions, any dissolved ReI is volatilized. In distilling
off the last of the ethel', care should be used so that the flask
will not be heated above 100C. because salicylic acid sublimes
slowly at higher temperatures.
2. "Add 2 ce. of neutral alcohol and 15 ce. of distilled water to the residue,
and titrate wiih tenth-normal sodium hydroxide, using phenolphthalein T.S.
as the indicator.
"Each cubic centimeter of tenth-normal sodium hydroxide is equivalent
to 0.01600 Gm. of sodium salicylate, C,H . OII.COONa.
"No'rE: This assay is applicable to ihci'ie Tablets when not coated.
Suitable modifications or another method IlJay be necessary for assayiIlg the
Tablets whcn coated."

N ClItral alcohol is added to dissolve the salicylic acid which is

but slightly soluble in water. Alcohol is frequently acid in
reaction and should always be neutralized if necessary, when
used as solvent in neutralization analyses.
C aH 4 (OH)COOH + NaOH--JoC eH 0HCOONa + H
4 20
138.05 160.04
One cubic centimeter of 0.1 N NaOH is equivalent to

10 1~8i~~00 = 0.013805 Gm. of salicylic acid

Questions and Problems

1. Write equations for the renetions that oeeur in this assay using struc-
tural formulae.
2. Ascertain the solubilities of salicylic acid in water and in ether. Explain
why the acid partitions itself largely into the ether layer.
S. What amounts of sodium salicylate might tablets labeled "5 grain
tablets of sodium salicylate" contain and meet the official tolerance
allowances?
110 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

4. From a bottle labeled "5 grain sodium salicylate tablets," 20 sample

tablets were removed and found to weigh 7.9246 Gm. A 0.3 Gm. sample
taken from the powdered tablets required 16.05 cc. of 0.1012 N NaOH.
Calculate the amount of sodium salicylate contained in each tablet :md the
per cent deviation of the tablets from the labeled amount.
Exercise 27
Object.-Assay of Tartaric Acid.
Materials Required.-3 Gm. of tartaric acid.
Normal sodium hydroxide.
Procedure.-" Place about 3 Gm. of Tartaric Acid in IL tared, glass-
stoppered flask and weigh accurately. Dissolve the Acid in about 40 cc.
of recently boiled distilled water, ILnd titrate with normal sodium hydroxide,
using phenolphthalcin T.S. as thc indicator. Each cubic centimeter of
norrnal sodium hydroxide is equivalent to 0.07503 Gm. of H 2C 4 H.0 6."

Phenolphthalein is the most suitable indicator to use in the

titration of organic acids. Tartaric acid reacts as a dibasic acid
as follows:
C 2H 2 (OH)2(COOHh + 2NaOH------7Na C. H 0 + 2H zO
2 I 4 6
150.05
Since 2 moles of NaOH are required to neutralize each lUole of
tartaric acid, each cubic centimeter of N NaOI-I is equivalent to

2, l~Oi~~() = 0.07503 Gm. tartaric acid.

Questions and Problems

1. Why is phenolphthalein employed as indicator in the titration of
organic acids with strong alkalies?
2. Write equations for theabove reaction structurally.
3. How much citric acid is equivalent to 1 cc. of 1.0250 N NaOH? Indi-
cate all calculations.
4. Write equations for the reactions that occur in the assay of; (a) lLC(ltic
anhydride, (b) tl'ichloroacetic acid, (c) sulfosalicylic acid, Cd) sodium bitar-
trate.
6. What would be thc titer of each of the above substances for 0.1256 N
NaOH?
RESIDUAL TITRATION METHODS
Exercise 28
Object.-Assay of Aromatic Sulfuric Acid.
!J.[aterials Required.-5 co. of aromatic sulfuric acid.
 ACIDIMETRY 111
Normal sodium hydroxide.
Normal sulfuric acid.
Procedure.-" Accurately measure 5 cc. of Aromatic Sulfuric Acid into a
tall bcaker. Add exactly 30 cc. of normal sodium hydroxide a.nd evaporate
the mixture to complete dryness 011 a. salld bath. Dissolve the residue in
30 ceo of distilled water, and titrate the excess of alkali with normal sulfuric
acid, using methyl orange T.S. as the indicator. Each cubic centimeter of
normal sodium hydroxide is equivalent to 0.04904 Gm. of H 2SO t ."
Aromatic sulfuric acid contains ethyl sulfuric acid, alcohol,
and volatile oils in addition to sulfuric acid. The mixture
after treatment with a measured excess of N N aOH is evaporated
to complete dryness on a sand bath to deeomposc any ethyl
sulfuric acid and to drive off the volatile ingredientH. The fn~e
alkali and sodium sulfate which remain in the residue are then
dissolved in distilled water and the excess alkali is determined
by titration with normal sulfuric acid, using methyl orange
indicator. The small quantities of alkali consumed by the
organic acids present in the volatile oil are negligible, since the
U.S.P. requirement is sufficiently broad to cover variation from
this source, being not less than 19 per cent and not more than
21 per cent of H 2S0 4
H 2S0 4 + 2NaOH~Na2S04 + 2H zO
98.08
Since 1 mole of H 2S0 4 requires 2 moles of NaOH, each cubic
centimeter of N NaOH is equivalent to 2 ~'fgoo = 0.04904 Gm.
H 2S0 4
Questions and Problems
1. Why is methyl orange used in place of phenolphthalein in this assay?
2. Why is the solution of aromatic sulfuric aeid evaporated to dryness on
a sand bath? Could a water bath be used?
S. Enumerate other official acidimetric assays in which residual titration
is employed.
Exercise 29
Object.-Assay of Tablets of Acetylsalicylic Acid.
Materials Required.-20 tablets of acetylsalicylic acid. (N OTE.-Two
5 grain tablets may be used for student work provided that the instructor
gives the weight of 20 tablets.)
0.1 N sulfuric acid.
0.1 N sodium hydroxide.
112 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

~~~2;~~g~~~~~~gg~~~~~8~~~
~OOO~~=o=m~=MM~~~~~m~om~~

~~88~~gb~8~g88~~soa~~~~8~
0000000000000000000000000

~~~~~~~~~~~~~~~~~~~~~~~~~
___
_rl~~OrlO~ __ _
~_~~rlOOOrl_rl

OOO~~OOOOOOooooo~omoooooo
~~~~~~~~~ci~~~~~~ci~~~ci~~~~
c-:t ...-I ........ _
 A(}JDIME1'RY 113

co
co ."
r~
o

.......... t:ll'"i
...... I"""'f r-l ~ C'.J P"'"f.-l
O}lDv.l~r-I-.:tIOe e-lIXlOo:.~
Cl':)tr.:l 000 CIJ"IflOO
.-t.,....: C'I COl 00 ...... r::'l ..-I
O_CO"l;tl~ :!!::!:
88~?:li:l
Cl Cl
OOOO ..... O.-tO 00
00000000 00000 00

\t:IlQlt:lr.oOOOl~
.
00
Oro
, ,
Ie N
ooooto:i""';e-ici 00 00

:~ ...'0~
m
0
~
.~
. "
',.c .",
~.;a~ :3
'Po

0
~
:",'" "
: .~
114 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Procedure.-1. "Weigh not less than 20 of the Tablets, reduce them to a

fine powder without an appreciable loss, and dissolve an aliquot portion,
representing approximately 0.7 Gm. of acetylsalicylic acid, in 20 cc. of
neutral alcohol in an Erlenmeyer flask, and titrate the solution immediately
with tenth-normal sodium hydroxide, using phenolphthalein T.S. as the
indicator."
Care must be observed in powdering the tablets because the
sample will not be representative if any of the material is lost.
The sample is titrated with sodium hydroxide to neutralize any
free acid formed by hydrolysis of the acetylsalicylic acid as
represented by the equation:
C aH4.0COCHa.COOH + HOH--l-CsH .OH.COOH + 4
CHaCOOH
and to neutralize the carboxy group of the acetylsalicylic acid:
C aH 4 .OCOCHa.COOH + NaOH----?C H .OCOCHaCOONa +
6 4
180.06 HOH
2. "Now add a volume of tenth-nOl"mal sodium hydroxide equal to that
used in the above titration plus 5 cc. more. Heat the mixture on a steam

TAJ3LE XI.-Ol!'FlCIAL AClDIMETRIC ASSAYS BY RESIDUAL TITRA'l'ION

Amount
Nor- Equh'll-
used, Indi-
Substance mality lent of Official requirement, per cent
am. or cator
of alkali 1 cc.
ce.
--- ---
U.S.P.
Acid, acetylsalicylic 1.5 Pp. 0.5 0.04502 C.H.IO(CR,COlCOOH = 9fl.5
Acid, lactic ........ 2.5 Pp. 1.0 0.09005 CH,CROHCOOH ~ 85 to no
Acid, sulfuric aro-
matic ........... 5.0 M.O. 1.0 0.04904 H2S0, = 19 to 21 W IV
Chloral hydrate ... 4.0 Pp. 1.0 0.1654 CCl,CHO.H20 = 99.5
Formaldehyde,
solution ... , .... 3.0 B.T.B. 1.0 0.03002 HCHO = 37
Methyl salicylate .. 2 PP. 0.5 0.07603 C,IhOH.CO,CH, = 98
N.F.
Acetylsalicylic acid,
tablets of .. , .... 0.7- Pp. 0.1 0.01801 C,H . OCOCH,.COOH =92.5
to 107.5'
Ethyl acetate ..... 1.5 ,.pp. 0.5 0.04403 CHaCOO.C,H. = 99
Formic acid, spirit
of .............. 10 Pp. 0.1 0.004602 HGOOH = 0.95 to 1.05W/V

B.T.B. = bromthymol blue. M.O. = methyl orange. Pp. = phenolphthalein .

Weight of ingredient SQught.
Per cent of labeled amount.
 ACIDIMETRY 115
bath for 15 minutes, with intermittent stirring, allow the solution to s.tand
at room temperature for 5 minutes, and then titrate with tenth-normal
sulfuric acid. Subtract the number of cubic centimeters of tenth-normal
sulfuric acid used, from the total number of cubic centimeters of alkali
added the second time, and multiply the result by 0.01801. This result
represents the weight of acetylsalicylic acid present in the portion of the
powder lIsed for the assa.)r."

Upon heating the titration mixture after the addition of an

excess of 0.1 N alkali, the ester is saponified:
C 6H4.0COCHa.COONa + Nn,oH-~C6H4.0H.COONa +
CHaCOONa + HOH
The amount of 0.1 N alkali remaining after the saponification
is complete is then determined by titration with 0.1 N acid.
QUestions and Problems
1. Show how the equivnll\nt fo~ acetylsalicyl.ic acid is derived.
 CHAPTER VII

- PRECIPITATION METHODS
Under Neutralization Methods, a class of reactions was con-
sidered which were of value in quantitative analysis because little
ionized substances 01' gases or both little ionized substances and
gases were formed. In the following exercises, a class of reactions
is dealt with which requires the formation of very slightly
soluble substances to cause the reactions to go to sufficient com-
pletion to be quantitative in nature. The solubility product
principle may be applied to all precipitation reactions, and it
should be reviewed before the subsequent assays are undertaken.
Determination of the End Point.-The end point of a reaction
in analyses by precipitation methods may be determined in one
of three ways:
1. By adding a standard solution to a solution of the substance
being analyzed until no. further precipitate is produced. This
method is sometimes applied in the determination of the chloride
ion content of chlorides with standard solution of silver nitrate.
2. By adding .the standard solution to a clear solution of the
substance being analyzed until a precipitate begins to form.
This method is often used in the titration of alkali cyanides with
standard silver nitrate solution. (See sodium cyanide reagent,
U.S.P. and diluted hydrocyanic acid, N.F.)
3. By means of ~ suitable indicator. All of the official assays
by precipitation methods employ indicator solutions to show the
end point of the reaction.
Indicators.-The indicators used in the official volumetric
precipitation assays are:
1. Ferric ammonium 8ulfate T.S. prepared by dissolving 8
Om. of reagent ferric ammonium sulfate in sufficient distilled
water to make 100 cc. This indicator is used in both direct and
residual tit rations of silver and mercury salts with standard
ammonium thiocyanate solution. The thiocyanate reacts with
116
 PRECIPITATION METHODS 117

the silver or mercury present to form a white precipitate of

silver or mercury thiocyanate but as soon as all of the silver
or mercury has been precipitated the thiocyanate reacts with
ferric ammonium sulfate to form red ferric sulfocyanate; the
first appearance of the red color marks the end point of the
reaction.
2. Potassium chromate T.,s. prepared by dissolving 10 Om. of
reagent potassium chromate in sufficient distilled water to make
100 cc. It may be used as indicator in the titration of chloride
solutions with standard silver nitrate solution, but in the Phar-
macopoeia it is employed only in the assay of oil of bitter almonds
for hydrocyanic acid. The end point is shown by the production
of a permanent red color caused by the formation of silver
chromate.
Standard Solutions.-The standard solutions employed in
the official assays by precipitation methods are tenth-normal
silver nitrate and tenth-normal ammonium thiocyanate.
Exercise 30
Object.-To Prepare and Standardize 0.1 N Silver Nitrate.
Materials Required.-lO Gm. of silver nitrate.
Procedure.-Dry about 10 Gm.' of pure silver nitrate in an electric oven
at 1100 0. until of constant weight. Weigh out exaetly 8.4945 Gm. of the
dried salt and dissolve it in sufficient distilled water to make 500 ce.

The pure analyzed silver nitrate now available on the market

makes it possible to prepare the solution directly from the
dried salt. Every precaution should be exercised in weighing
the silver nitrate, since a slight error in the weight would make
the concentration of the solution inaccurate.
Mol. wt. AgNO a = 169.89
Therefore 1,000 cc. of 0.1 N AgNO a requires 169.89/10 =
16.989 Gm., and 500 cc. of 0.1 N AgNO a requires 16,989/2 =
8.4945 Om. pure AgNO a
If the purity of the silver nitrate crystals used is doubtful, the
concentration of the volumetric solution may be determined by
titration against an accurately weighed sample of pure sodium
chloride in the same manner as given under the assay of sodium
chloride.
118 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The distilled water should be tested for halides as followR

before it is used to prepare standard Hilver nitrate solution:
Fill a test tube three-fourths full of water, add about 0.5. cc.
of silver nitrate solution Hnd a few drops of U.S.P. nitric acid.
Any turbidity is positive evidence that the water contains
halides and is unfit for use in preparing the standard solution
unleso; the concentration of the solution is to be determined by
titration against a pure chloride.

Exercise 31

Object.-To Prepare and Standardize 0.1 N Ammonium

Thiocyanate.
Materials Required.-4.0 Gm. of !nnmOniulll thiocYlLllate.
0.1 N silver nit.rate.
Procedure.-Dissolve 4.0 Gm. of reagent 1ll111110uilllll thiocyanate in 500
eo. of distilled water. Ascertain its strength by titration against tenth-
normal sih~er nitrate, and adjust to exact normality or to a known strength
of approximate normality.

Since ammonium thiocyanate is a deliquescent salt, a slight

amount in excess of the theoretical quantity required is used to
prepare the solution. The normality of the solution is deter-
mined by titration with 0.1 N AgNO a as follows:
Measure exaetly about 30 ec. of 0.1 N AgNO a into a 250 ce.
Erlenmeyer flask, and dilute the solution with 50 ec. of dis-
tilled water, add 2 cc. of ferric ammonium sulfate T.S, and
2 cc. of nitric acid to prevent hydrolysis of the ferric alum,
and titrate the mixture with the approximately 0.1 N NH 4SCN
solution .. When the thiocyanate solution is run in, a white
precipitate is produced giving the mixture a milky appearance,
and as the thiocyanate is added drop by drop, it produces a
brownish-red cloud which quickly disappears on shaking. As the
end point is reached, the precipitate becomes flocculent settling
readily and, finally, a drop or two of thiocyanate solution pro-
duces a permanent reddish color which does not disappear on
shaking.
Ammonium thiocyanate solution added to an acidulated solu-
tion of silver nitrate containing ferric alum precipitates silver
thiocyanate until all of the silver has been precipitated, then the
 PRECIPITATION ME7'HODS 119

slightest excess of ammonium thiocyanate solution reacts with

ferric alum to form red ferric thiocyanate as fo11o\vs:

AgNO a + NH SCN-AgSCN + NH NO a
4 4
169.89 76.11 white
2FeNH4(SO~h + 6NH 4SCN--l-2Fe(SCN)a + 4(NH )2S04
4

red
If 30.00 cc. of 0.1 N AgN0 3 require 28.13 cc. of NH 4SON solu-
tion, the factor of the NH 4SCN solution ill terms of 0.1 N is
30.00/28.13 = 1.0666.
Oxides of nitrogen yield colored salts with ferric filum. Solu-
tions containing oxides of nitrogen, therefore, Bhould be boiled
prior to the addition of the indicator.

"Questions and Problems

1. Explain in detail how to standardize a solution of silver nitrate against
pure sodium chloride.
2. Why is nitric acid added when NH 4SGN solution i~ stlmdardizeti
against 0.1 N AgNO, using ferric alum as indicator?
3. Write ionic ally the reactions involved in the standardization of
NH.SGN against AgNO, with ferric alum indicator.
4. What must be the normality of solution of silver nitrate so that each
cubic centimeter shall be equivalent to 6 mg. of NaG!?
5. How might 0.1 N AgNO a be prepared from pure metallic silver?
6. Why should standard silver nitrate solution bc protected from strong
sunlight?
7. Why must oxides of nitrogen be ahHent in titration~ with NH.SGN?
8. How much 0.1 N AgNO a could be prepared frOlll 5 Gm. of pure silver?
9. To how much Hg, Ag, toughened AgNO., and HgO is 1 cc. of 0.1000
N NH,SGN equivalent?
10. If 1 ce. of a solution of silver nitr:tte is equivalent to 0.0148 Gm. of
NaGI, what is the normality of the silver nitrate solution?

DIRECT TITRATION METHOD

Oompounds of silver and mercury which can be readily con-
verted into silver nitrate and mercuric nitrate, respectively,
may be estimated by direct titration with standard potassium
thiocyanate solutioll, using ferric ammonium sulfate as indicator.
The method is based on the quantitative precipitation of the
corresponding thiocyanate, e.g.:
120 QUANTITATIVE PHARMACEU'l'ICAL CHEMISTRY

and

When all of the metallic ion has been precipitated as thio-

cyanate, the NH 4SCN reacts with the ferric alum indicator to
form red-colored ferric thiocyanate, marking the end point of
titration:
2FeNH 4 (S04)2 + 6NH4SCN~2Fe(SCN)3 + 4(NH )2S04 4

The solution must be acidulated with nitric acid to prevent the

hydrolysis which ferric salts undergo in neutral Rolution. Chlo-
rides must be absent, since the chlorides of silver and mercury
are more soluble than the respective thiocyanates. This
method of assay is exemplified in the following exercise:

Exercise 32
Object.-Assay of Strong Silver Protein.
Materials Required.-2 Gm. of strong silver protein.
About 10 cc. of nitric acid.
2 cc. of ferric alum indic~Ltor (8 Gm. FcNH,(SO')2.12H,O in 100 cc.).
0.1 N ammonium thiocyanate.
procedure.-l. "Ignite about 2 Gm. of Strong Silver Protein, accurately
weighed in a porcelain crucible until nIl of the carbon is burned off. Trans-
fer as much as possible of the residue to a beaker, add to the crucible 5 ceo of
nitric acid, warm to dissolve any adhering silver, and transfer the solution
1,0 the beaker with the aid of a little distilled water. Cover the beaker, and
heat on a water bath unW all of the metallic silver is dissolved, l1elding a
little more nitric acid, if necessary. II

Upon ignition, strong silver protein yields a residue of metallic

silver and silver oxide. The silver and silver oxide are dissolved
in nitric acid:

Ag + 2HN08~AgN03 + H 20 + N0 2
Ag 20 +
2HN03~2AgN03 H 20 +
2. "Filter into a flask, wash the insoluble residue thoroughly with dis-
tilled water, cool, and dilute with distilled water, if necessary, to about
75 ceo Add 2 ceo of ferric ammonium sulfate T.S., and titrate with tenth-
nonnal ammonium thiocyanate Each cubic centimeter of tenth-normal
ammonium thiocyanate is equivalent to 0.01079 Gm. of silver."
 PRECIPITATION METHODS 121

The mixture is filtered to free it of carbonaceouR matter,

the residue on the filter is washed free of silver nitrate, and the
nitrate is titrated with 0.1 N NH 4SCN:
AgNO a + NH 4SCN-)AgSCN + NH 4NOa
169.89
Each cubic centimeter of 0.1 N NH 4SCN is equivalent to
107.88
10 X 1000 = 0.01079 Gm. Ag. The U.S.P. requires that strong
silver protein contain 7.5 to 8.5 pel' cent silver. Calculate the
percentage of Rilver in the sample analyzed and compare your
result with the U.S.P. requirement.
Questions and Problems
1. How does the Rolubility proouct prineiple apply in the above assay?
2. If each cuhic centimeter of Il solution of NHjSCN is known to be
TABLE XII.-OF~'lCIAL SUBSTANCES ASSAYED BY DIRECT TITIlATION WITH
0.1 N NH.SCN

Amount Equiv!1-
Official require-
Substance used, Gm. lent of
ment, per cent
or ce. 1 ce.

U.S.P.
Mass of mercury ............. . . 0.5 0.01003 Hg = 32 to 34
Mercuric oxide, yellow ........ . 0.5 0.01083 HgO =99.5
Mercuric salicylate ........... . 0.5 0.01003 Hg = 54 to 59.5
Mercuric sllcciniinide ......... . 0.5 0.01003 Hg = 49.5 to 51
Mercury, oleate of ............ . 0.75 0.01083 HgO = 24 to 26
Mercury ..................... . 0.4 0.01003 Hg = 99.5
Mercury with chlllk ........... . 1.0 0.01003 Hg ~. 37 to 39
Ointment, mercurial, mild ..... . 1.0 0.01003 Hg = 29 to 31
Ointment, mercurial, strong ... . 1.0 0.01003 Hg = 49 to.51
Silver nitrate ................ . 0.8 0.01699 AgNO. = 99.8
Silver nitrate, toughened ...... . 0.8 0.01699 AgNO. = 94.5
Silver protein, mild ........... . 1.0 0,01079 Ag = 19 to 25
Silver protein, strong ......... . 2.0 0.01079 Ag = 7.5 to 8.5
N.F.
Arsenic and mercuric iodides,
solution of (for HgI 2) . 25 0.02272 Hgh "" 0.95 to 1.05
Mercuric oxide, red. . ........ . 0.5 0.01083 HgO ""99.5
Mercuric salicylate, ampules of o.5a 0.01003 Hg = 51.0 to 62.46
Mercuric succinimide, ampuls of 0.25 a 0.01003 Hg = 47.7 to 52,8"

Amount of ingredient Bought.

" Per cent of labeled ~mount.
122 QUANTI'l'ATIVE PHARMACEUTICAL CHEMISTRY

equivalent to 0.01140 Gm. of pure Ag, what would be its equivalent of I-Ig,
HgI2' HgO, Hg(NO,)2, AgN0 3, and Ag,O?
3. Give a method for the assay of metallic mercury. Explain each step
in the procedure and write equations for all reactions.
4. What must be the normality of a solution of NI-I 4SCN so that each
<mbic centimeter shall be equivalent to 1 mg. of silver?
5. Write equations for the reactions that occur in the assay of: (a) yellow
mercuric oxide, (fJ) mercuric succinimide, (c) mercuric iodide in solution of
ai'senic and mercuric iodides.

RESIDUAL TITRATION METHOD (VOLHARD'S METHOD)

. This method is based on the complete precipitation of insoluble
silver salts from nitric acid solution by the addition of excess
standard silver nitrate solution to a soluble salt and the deter-
mination of the amount of silver nitrate solution in excess by
residual titration with standard ammonium thiocyanate c.:olution,
using ferric ammonium sulfate as indicator.
Exercise 33
To Determine the Purity of a Soluble Chloride.-The purity of
any soluble chloride may be determined by the method given
below, provided that other substances which give precipitates
with silver nitrate are absent. Substances ot11er than chlorides
which produce slightly soluble silver salts are bromides, iodides,
cyanides, sulfocyanides, sulfides, phosphates, arsenates, car-
bonates, etc.
Object.-Assay of Sodium Chloride.
Materials Required.-1 Gm. of sodium chloride.
5 cc. of nitric acid.
2 ee. of ferric alum indicator.
50 ce. of 0.1 N silver nitrate.
0.1 N ammonium thiocyanate.
Procedure.-" Dry about 0.25 Gm. of Sodium Chloride to constant weight
at 100C., weigh accurately, and dissolve in 50 ce. of distilled water in a
200-cc. volumetric flask. Add 50 ce. of tenth-normal silver nitrate to the
solution, then l5 ce. of nitric acid, and sufficient distilled water to make the
mixture measure 200 cc. Mix well, filter into a dry flask through a filter
which has not been moistened, and reject the first 20 cc. of filtrate. To
exactly 100 cc. of the subsequent filtrate add 2 ce. of ferric ammonium sulfate
T.S., and titrate with tenth-normal ammonium thiocyanate. Eaeh cubic
centimeter of tenth-normal silver nitrate is equivalent to 0.005845 Gm. of
NaC!."
 PRECIPIT A'l'ION ME'l'HODS 123
Nitric acid is added to prevent the precipitation of silver as
carbonate, phosphate, etc. and to prevent hydrolysis of the ferric
alum.
The precipitated chloride is removed by filtration, since
silver chloride is several times more soluble than silver thio-
cyanate. If the precipitated silver chloride were left in the
titration mixture when the excess silver nitrate is titrated with
ammonium thiocyanate, the following reaction would take place
between the dissolved AgCI and NH 4SCN: 9

AgCI + NR1SCN---}AgSCN + NH CI 4

More precipitated AgCI would then dissolve to re-establish

equilibrium between the precipitate and solution. This would
make the end point uncertain.
The residual liquid after precipitation, therefore, is made up
to a definite volume, 200 cc., and an aliquot portion, 100 ce., of
the filtrate is taken for residual titration. The first 20 (Je.
of filtrate is rejected to avoid possible error due to absorption of
silver nitrate by the filter. The aliquot-portion method elimi-
nates the tedious procedure of washing the precipitate free of
silver nitrate whieh wuuld be required if all of the excess silver
nitrate were to be titrated residually.
NaCl + AgNOs--AgCl + NuNO s
58.45
AgNO a + NH,jSCN----+AgSCN + NH 4NO a
Each cubie centimeter of 0.1 N AgNO a i:-; equivalent to
58.45
- - - -.... _- = 0 005845 Gm. N aCt
10 X 1000 .
The number of cubic centimeters of 0.1 N NH 4SCN required in
the residual titration is multiplied by 2 because only one-half the
excess 0.1 N AgNO s is represented in the aJiquot portion.
[cc. 0.1 N AgNOa - (2 X ee. 0.1 N NH 4SCN)] X 0.005845
\Vt. of sample
X 100 ::= per cent purity of NaCl
Questions and Problems
1. Why is nitric acid added in the above assay?
2. Why must the precipitated AgOl be removed before titration with
stb.udard NH.SON? Explain in detail.
124 QUANTITA'1'IVE PHARMACEU'1'ICAL CHEMISTRY

3. How might all of the excess silver nitrate solution be freed from the
precipitate and titrated?
4. Name several official chlorides assllyed by the above method.
5. What must be thc normality of a standard silver nitrate solution so
that each cubic centimeter will be equivalent to 1 mg. of chlorine?
Exercise 34
Object.-Assay of Ammonium Bromide.
Materials Requited.-OA Gm. of ammonium bromide.
2 cc. of nitric acid.
2 cc. of ferric alum indicator.
50 cc. of 0.1 N silver nitrate.
1J.1 N ammonium thiocyanate.
Procedure.-"Dry about 0.4 GIn. of Ammonium Bromide to constant
weight in a desiccator over sulfuric acid, and weigh accurately. Dissolve
it in about 50 cc. of distilled water, add 50 cc. of tenth-norlllal silver nitrate,
2 cc. of ferric ammonium sulfate T.S., and 2 cc. of nitric acid. Titrate the
excess of silver nitrate with tenth-normal ammonium thiocyanate. Each
cubic centimeter of tenth-normal silver nitrate is equivalent to 0.009796 Gm.
of NH4Br. Each gram of Ammonium Bromide, previously dried, is equiv-
alent to not less than 101.1 cc. and not more than 103.0 of tenth-normal
silver nitrate."

The above assay is similar to that for chlorides, except that the
precipitated silver bromide need not be removed by filtration,
since it is less soluble than silver thiocyanate.
The reactions and calculations are similar to those of the pre-
ceding assay .. Write equations for all reactions involved and
calculate the per cent purity of the ammonium bromide.
Questions and Problems
1. How does the prescnce of excess silver nitrate tend to produce quanti-
tative precipitation 'of silver bromide?
2. How does the formation of insoluble silver bromide force the reaction
to completion?
3. Why may the residual titration of excess silver nitratc be conducted
without removal of the precipitated silver bromide?
4. Look up thc solubility product of AgBr and calculate the solubility of
silver bromide. .
5. Write all equations ionically.
Exercise 35
Object.-Assay of Syrup of Hydriodic Acid.
Materials Required.-25 ce. of syrup of hydriodic acid.
40 ec. of 0.1 N silver nitrate.
 PRECIPITATION METHODS 125
. 5 ce. of nitric acid.
2 cc. of ferric alum indicator.
0.1 N ammoniuin thiocyanate.
Procedure.-"Plaee exactly 25 ee. of Syrup of Hydriodic Acid in a flask,
dilute it with 100 ee. of distilled water, add 40 cc. of tenth-normal silver
nitrate, agitate the mixture, add 5 ce. of nitric acid and heat the mixture on
a water bath until the precipitate has acquired a hright yellow color. Cool,
add 2 ce. of ferric ammonium sulfate T.S., :lnd determine the residual silver
nitrate by titration with tenth-normal ammonium thiocyanate. Ea('h eubic
centimeter of tenth-normal silver nitrate is equivalent to0.01279 Gm. of HL"

This assay is similar to that for bromides exeept that a

measured volume of liquid is used for the sample.
HI +
AgNOa--+AgI + HNO a
127.93
~gN03 + NHojSCN--+AgSCN + NH 4 NO a.

Calculate the amount of HI equiyalent to 1 cc. of 0.1 N AgNO a

and the per cent HI in the sample analyzed. The U.S.P.
requires that syrup of hydriodic acid contain not less than 1.3
and not more than 1.5 Gm. of HI in each 100 ce. Note that
this is a weight-to-volume percentage.
Questions and Problems
1. Why must a large amount of nitric acid be avoided in the precipitation
of chlorides, bromides, and iodides as the eorrcHponding Hilver salts?
2. If 25 cc. of syrup of hydriodic aoid containing 1.4 per cent HI is
treated with 40 ce. of 0.0955 N AgNO" how much 0.1225 N NH.SCN will be
necessary to titrate the excess AgN0 3 ?
3. Write equations for the reactions that occur in the assay of ((11 chloro-
form liniment, (b) potassium nitrate, (e) syrup of ferrolls iodide, and (d)
acriflavine.
4. A solution is known to contain 5.3490 Gm. of silver combined as nitrate
per 1,000 ee. How much of each of the following substances would be
equivalent to 1 cc.: Br, CaCh, Fer" HCI, NaI, NaCN, and NHICl?
Exercise 36
Object.-Assay of Elixir of Three Bromides.
Materials Required.-l0 {c. of elixir of three bromides.
0.1 N silver nitrate.
2 ec, of nitric acid.
2 cc. of ferric alum indicator.
0.1 N ammonium thiocyanate.
126 QUANTITATIVE PHARMACEU'PICAL CHEMISTRY

Procedure.-" Dilute exactly 10 ce. of the Elixir with distilled water to

250 ce. To 25 ee. of the dilution, add slowly and with agitation, 50 ee. of
tenth-normal silver nitrate, 2. ce. of nitric acid, and 2 ee. of ferric ammonium
sulfate T.8. Titrate the excess silver nitrate with tenth-normal ammonium
thiocyanate.
Each cubic centimeter of tenth-normal silver nitrate is equivalent to
0.01059 Gm. of the total bromides."

In calculating the amount of bromide equivalent to 1 cc. of

0.1 N silver nitrate in elixir of three bromides, elixir of five
bromides and syrup of five bromides and similar mixtures the
following general formula may be used:

where F = number of grams of mixed bromides equivalent

to 1 cc. 0.1 N AgNO a
a = total number of grams of bromides (based on
100 per cent purity) in mixture.
bl , b2, ba, etc. = number of grams of individual bromides (100
per cent pure) in mixture.
etc. = number of grams of the bromi.de represented by
Gl, C2, Ca,
bl , b2 , bs, etc., equal to 1 cc. 0.1 N AgNO a.
The factor for elixir of three bromides, which contains 80 Gm.
of sodium bromide, 80 Gm. of ammonium bromide and 80 Gm.
of potassium bromide per 1,000. cc. of solution, is calculated
thus (assuming 100 per cent purity):
240
F = 80 80 80 = 0.01059
0.010291 + 0.011901 + 0.009796
The factor for syrup of five bromides may be calculated as
follows: The syrup contains 80 Gm. of potassium bromide, 80 Gm.
of sodium bromide, 50 Gm. of ammonium bromide, 25 Gm. of
calcium bromide and 8 Gm. of lithium bromide per 1,000 cc.
of solution. If these substitutions are made in the formula,
237.8
F = -~80;;---._.;.___---;:;:8o;;-------,;;50,,---.2:;-;;5~X:-;0~.8~4;---;8:;-:-X:-0;;:-.=85
0.011902 + O~OT0292 + 0.009796 + 0.009996 + 0.008686
= 0.01058
 PRECIPITATIOJv lvIETHODS 127
TABLE XIII.-OFFICIAL SUllBTANC'ES ASSAYED BY RESIDUAL TITItATION OF
0.1 N AgN0 3 WITH 0,] N NH 4SCN

Amount Equivll-
Jeno of
used, Ofllcialrequirement,
Substance 1 CC. of
GlII. per cent
0.1 N
or co.
Nfl,SCN

U.S.P.
Acid hydriodic. diluted ...... " .. 5 0.01279 HI = 0.[, to 10,5
_-\.t:riflavine ..... , ..... , ........ . 0.25 0.003546 CI = 13.3 to 14.3
Acriflavine hYdrochloride ....... . 0.25 0.00354" Cl = 23 to 24,0
Arnmonium bromide ... , ....... . 0.4 0.009796 NH.,EI' = 90
AmIllonium chloride ............ . 0.1 0.005350 NH,Cl = 99.5
Calcium cbloride. R ............ . 2 O. 005550 CaCh = 74
Chlorofol'ln, liniment of. ..... " .. 10 D.OD39S CHCI, = 40 to 4,'; W/V
Chloroform, spirit of. .......... . I) 0.00398 CHC]' = 8.5 to G.25W/V
Oil of musturd, vobtile ...... " .. 4 0.004956 C.H,NCS = 93
Potassium bromide . ............ . 0.4 0,01190 KBr = 99
Potassium nitrute .. , ........ " .. 0.4 0.01011 KNO, = 1J9
Sodium bromide ............... . 0.4 0.01029 NnBr = 99
Sodi um en loride ............... . 0.25 0.005845 NaCI = 99.5
Symp of ferrous iodide ......... . 10 0.01548 Fer, = 0.5 to 7.5W/V
Syrup of hydriodic acid. . . . . . . . .. 25 0.01279 HI = 1.3 to 1.511' IV
N.F.
Ammonium bromide, elixir of. .... 10 0, 000796 NH.,Hr = 8 til 9 IVi V
Ammonium chloride, tablets of ... O. 00535 NH,Cl = 92.5 to 107.5'
Ammonium iodide ..... , .... " .. 0.5 0.01450 NH,I = 98
Bromides, five, elixir of ....... , .. 10 0.0102 total bromides = 25 to 27W/V
Bromides, three, elixir of. . . . . . . .. 10 0.01059 total bromides = 23 tu 25W/T"
Bromides, three, tablet. of. ... , .. 0.6" 0.007992 Hr = 70 to 81 b
Lithium bromide ........... ,., .. 0.35 O. 008686 LiHr = 8" to 1)0
Potas..'3iuTIl onlluide, elixir of .. . , . , 5 0.01100 KBr = 17 to 18W/V
Pata,8Hium chlorhle........ .. .. .. 0.25 0.OU7455 KCl = 91)
Potassium thiocyanate ........ ,.. 0.2 0.009716 KSCN = 99
Sodium bromide, elixir of. ... , .. , 5 0.01029 NaBr = 17 to 18 W IV
Sodium bromide, tablets of....... 0.3a O. ()1029 NaBr = 92.5 to 107.5.
Sodium chloride in isotonic solu-
tion of dextrose und ... ",..... 0.2" 0,005845 N"Cl = 0.4 to 0.425W/V
Sodium chloride in umpuls of dex-
trose undo .. ..... _ ... , ...... . 0.2,1 0.000S45 N "Cl = \l5 to 105~
Sodium chloride. ampul, of, , . , .. . 0.2" 0,005845 N act = 95 to 105'
Sodium nitrite, tablets of. ...... . 0.5(1. 0.02070 NI1NO. = 01 to 109'
Sodium thiocyllnate ........ ' " ., 0.2 0.008107 NuSON "" 98.5
Strontium bromide ............. . 0.5 0,01778 SrBn.OH,O = 98
Zinc iodide, .... , .......... , . , , . 0.5 0.01590 ZnIo = 98

R. reagent .
1'::1

Amount of ingredient sought.

b Per cent of the labeled amount.
128 QUANTITATIVE PlJARMACEUTlC'AL CHEMISTRY

According to the U.S.P. XI, calcium bromide should contain

at least 84 per cent of CaBr2, thus we have to multiply 25 Om. by
84 per cent to obtain the amollnt of 100 per cent CaBr2 present
The same applies to lithium bromide; the N.F. VI allows a
minimum of 85 per cent LiBr.

Questions and Problems

1. Show how the total bromide equivalent in the assay of elixir of five
bromides is derived.
2. Tablets of three bromides each weighing on the average 0.4252 Gm.
required 37.50 ceo of 0.1 N AgNO, in the assay of a powdered sample of the
tablets weighing'O.4012 Gm. Calculate the per cent of bromine and of total
bromides in the tablets.
3. Calculate the weight of hromine and of total bromides in each tablet
from the data given in probl~lll 2.
4. If the tablets assayed as in problem 2 were labeled ,e 5 grain tablets of
three bromides," would they conform to tho official Tequil'oments?
5. Calculate the minimum and maximum amounts of total bromides that
the tablets labeled as in question 4 might contain and meet the official
requirements.
 CHAPTER VIII

OXIDATION -REDUCTION METHODS

The chemical reactions whieh occur in neutralization and
precipitation methods of analysis take place without change in
valence. Oxidation-reduction methods of analysis nearly always
involve a change in valence of the reacting substances. A study
of the principles underlying the latter type of reactions is an
essential preliminary if the methods based on them are to be
understood.
Theory.-The simplest type of oxidation-reduction reMtions
are addition reactions. Thus, when oxygen gas unites with
hydrogen gas, forming water, O2 + 2H 2-2H 20, the oxygen
is reduced and the hydrogen oxidized. When carbon burns in
the presence of oxygen to form carbon dioxide, C + 02-----'>C0 2,
it is oxidized and the oxygen is reduced, and when carbon unites
with sulfur to form carbon disulfide, C + 2S---}CS 2, the carbon
is oxidized and the sulfur reduced.
From the above simple illustrations, it is obvious that reduction
need not imply a reaction of hydrogen, since oxygen is reduced
by carbon, and that oxidation need not imply a reaction of
oxygen, since carbon is oxidized by sulfur. When one sub-
stance is oxidized, some other substance must be correspondingly
reduced, and conversely, when one substance is reduced, some
other substance must be correspondingly oxidized.
The oxidation-reduction reactions which take place in the
official assay processes are, for the most part, between electrolytes
in aqueous solution. Their quantitative value is based upon the
fact that metals, non-metals, and their iOllS, under suitable
conditions, can be made to undergo a change in the quantity
of electric charge associated with them and that in the change
there exists a simple relationship between the quantity of
electricity lost or gained and the weight of the reacting sub-
stances. This relationship may be expressed according to Fara-
129
130 QUANTITATIl'E PHARMACEUTICAL CHEMISTRY

day's law as follows: A change of charge of one is equivalent /0

the gain 01' loss of 96,500 coulombs of electTicity for each formula
weight of element 01' group of elements involved. Since in every
oxidation-reduction reaction the charge lost or g!1ined by one
subst!j.1lCe must necessarily be gained or lost by another, it follows
that there is always a tran:-;fer of electricity in oxidation-reduction
reactions.
It has been shown by experimcnt that substanceH eIassified
chemically as reducing lose negative charges or electrons when
oxidized and that those classified as oxidizing gain electrons
when reduced. Oxidation, therefore, consists of the loss of
negative charge, and reduction consists of the gain of negative
charge. A unit negative charge is an eleclr'on according to the
electron theory of matter, and a unit positive charge, the proton,
is the charge left on an atom when 1:t has lost an electron. OX1:dation
may then be defined as the loss of one 01' more electrons by an atom
01' ion, and reduction may be defined as the gain of one 01' more
electrons by an atom or ion.
When metallic iron is oxidized to the ferrous ion, each atom of
iron loses 2 electrons. If an electron is represented by the nega-
tive sign (-), and the charge left on the atom by the positive
sign (+), the change may be indicated as Fe----)Fe++ + 2( -).
Thus, when iron wire is dissolved in hydrochloric acid the
following reaction occurs:

(1) Fe + 2HC1----)FeCb + H2
or
(2) Fe + 2H+ + 2Cl-----)Fe++ + 2Cl- +
H2
Zero + 2( +) + 2( -) = 2(+) + 2( -) + zero.
Since C1- is present on both sides of equation (2) the essential
reaction is Fe + 2H+----)Fe++ +
H 2 During the reaetion,
the iron has changed (been oxidized) from a neutral atom to an
iOll bearing two positive charges through the lOHH of 2 electrons,
and the 2 hydrogen ions have each gained 1 electron (been
reduced to molecular hydrogen).
If the ferrous ion is further oxidized to the ferric state, it
loses another electron:
Fe++ __ Fe+++ + (-)
 OXIDATION-REDUCTION METHODS 131

In the reaction
2Fe+1- + Ck--..72Fe+++ + 2Cl-
each ferrous ion loses I electron which is gained by a chlorine
atom, for a negative charge on an ion indicates that it hat; gained
an extra electron. In the above case, the-ferrous ion has been
oxidized, since it lost an electron; and chlorine has been reduced
to chloride ion, since it gained an electron.
When ferric chloride is reduced by stannOllS chloride, the
following reaction takes place: 2FeCI a + 811C1 2 -72FeC1 2 +
8nC14, which written ionically becomes
2Fe+++ + Sn++----t2Fe++ + Sn++++
2(55.84) 118.7

Each ferric ion gains one electron at the expense of the stannous
ions, each of which loses two electrons. The quantity of electric-
:ty gained by the iron is 96,500 coulombs for every 55.84 Gm. of
iron reduced, and that lost by the tin is 2 X 96,500 coulombs
for every 118.7 Gm. of tin oxidized. The chloride ion is in the
'lame state of oxidation before as after the reactioll, sillce it
undergoes no change of charge.
In most cases, the change of charge which an atom, iOll, or
raelieal undergoes in an oxidation-reductiOll reaction is llumeri-
cally equal to the change in valence, ~:.c.,'
1. When ferrous iron is oxidized to the ferric conditioll, the
charge associated with the iron atom changes from two to three:
2Fe++ + 2H+~2Fe+++ + H2
and the valence also changes from two to three:
Cl C1
/ /
2Fe + C12->2Fe-Cl
"Cl "Cl

2. When potassium pennanganate is i'educed, the manganese

atom gains five negative charges:
Mn 7(+)~ Mn++ + 5( +)
132 QUL1N'l'!1Wl'lVE PHARMACEU'l']CAL ClllJ,'MISTRY

and it changes from a valence of seven to a valence of two:

O-I<:
~O
Mn (See assay of FeS04 page 136.)
'\.~
~O
o
3. When potassium chlorate is reduced, however, the chlorine
atom changes from a charge of 5( +) to a charge of 1 ( - ), and
since the total change of polarity is thc algcbraic sum of the
charge carried by the atom on ench side of the equation, the
change in polarity of the chlorine atom is six:
C15(+l--tCI-
but the change in valence experienced by the chlorine is four:
O-K
/
CI=O --tKCl (See assay of KelO a, page 149.)
~
o
Standard Solutions.-The standard solutions used in the
official assays by oxidation-reduction methods are: potassium
permanganate, potassium dichromate, iodine and bromine as
oxidizing agents, and oxalic acid and sodium thiosulphate as
reducing agents.
A normal solution of an oxidizing or reducing agent is one which
contains lin moles of reagent per 1,000 ceo where n represents the
change of charge which the reagent undergoes in the reaction.
The change of charge which oxidizing agents ill the official
standard oxidizing solutions undergo is given below:
1. Pel'manganate ion.---7manganous ion
Mn04":' + 8H+-7Mn++ + 4H 20
1(-) + 8(+)-72(+) + zero + 5(+)
When the manganate ion is reduced to the manganous ion,
it gains five electrons or loses five positive cha,rges. A normal
solution of potassium permanganate should therefore contain
H mole of KMn04 per 1,000 cc.
 OXlDA'l.'IVN-REDUCTIDN };[ E'l'llOD8 132
2. Dichromate ion-chromic ion
C1'207 --+ 14H+-2Cr+++ 7H 20+
2( -) + 14( + )--76 ( +) zero + + 6( +)
Upon reduction to ('.hromic iOll, each dichromate iun loses
six positive charges. A normal solution of didll'ol11:lte must
therefore contain }li mole of K2Cr207 per 1,000 ee.

3. Iodine-)iodide ion
I2 ~ 21-
Zero 2(-) 2(+)+
Each atom of iodine gains 1 electron and supplies one positiy(~
charge. A normal solution would therrforc contain 1 mole of
iodide ion per 1,000 cc.
The change of charge which the reducing agents in the official
'lo
standard solutions undergo is as follows:

1. Oxalate ion-}carhon dioxide

C 204-- --,) 2C0 2 +
2( -)
2(-) zero +
2(-)

The change of charge is two pel' mole of oxalate ion, and a

normal solution of oxalic acid should contain 12 mole of oxalic
acid per 1,000 cc.

2. Thiosulphate ion--Jotetrathionatc ion

2S 2 0 3--- - 8 40 0- -
4(-) 2(-) + 2(-)
The change of charge is aIle pel' mole of thimmlpbate iOll, and a
normal solution of sodium thiosulphate should contaiu one mole
of sodium thiosulphatc per 1,000 cc.
Permanganate Methods.-Potassium permanganate volumet-
ric solution can he standardized easily, and it retains its COll-
centration over long periods of time when proper precaution&
are observed in its preparation and preservation. The reactions
of pennanganate in solution are rapid. It also serves as an
indicator in titrations where it is used, since a very slight excess
of permanganate imparts to solutions a distinct pink color.
134 QUA.NTITATIVE PHARMACEUTICAL CHEMIS1'RY

Exercise 37
Object.-To Prepare and Standardize 0.1 N Potassium
Permanganate.
Materials Required.-About 3.3 Gm. of potassium permanganate.
About 1 Gill. of reagent sodium oxalate.
Procedure.-l. "Dissolve ahout 3.3 Gm. of potas8iuIIl permanganate in
1000 ce. of distilled water iu a fla~k and boil the solution for about fifteen
minutes. Stopper the flask and allow it to stand for at least two days before
filt.ering through l1;;bestos."

Any organic matter which may be present in distilled water is

decomposed by potassium pennanganate. Consequently, after
solution of the pel'manganate is effected, the liquid is allowed to
s/;and for two days to insure the completion of the decomposition
reaction. The solution is then filtered through asbestos to,
remove all tl'ltces of manganese dioxide which if present acts as a
catalyst to promote the formation of more mtnganese dioxide at
the expense of permanganate ion.
2. "Ascertain its exact nonmdity by titration against reagent sodium
oxalate as follows:
"Weigh accurately about 0.2 Gm. of reagent sodium oxalate, previously
dried to constlLllt weight at llOC., and dissolve it in 250 ce. of distilled
water. Add 7 cc. of sulfuric acid, heat to about 70C. and then slowly add
the pcrmunganate solution from a burette, with constant stirring, until a
pale pink color is produced which per~ists for fifteen seconds. The tem-
perature at the conclusion of the titration should not be less than 60C.
C!Llculate the norm[Llity of the solution and, if desired, adjust the solution
exactly to tenth-normal."

Titrate the mixture by adding the permanganate solution

from a glass-stoppered burette, with continuous stirring of the
titration mixture until a drop of trw pormanganate causes a
permanent pink color. Read the top of the meniscus in the
burette instead of the lowest part. Using the first sample as a
preliminary !'Un, titrate the two other portions of oxalate in the
same manner and base the calculations of normality upon them.
Sodium oxalate is the best standard to use in the standardiza-
tion of potassium permanganate, since it can be obtained in
very pure condition. Sufficient sulfuric acid must be added to
keep the hydrogen ion concentration sensibly constant through-
out the titration, thereby preventing the formation of manganese
 OXIDATION-REDueTJON jly[ETFIODS 135

dioxide and supplying the hydrogen iOllS used up in the reduction

of the permanganate ion. Thus for every mole of pcnnangnnate
ion reduced, 8 moles of hydrogen ion are reqllil'f~d as "hmvn in the
equation below. The top of the meniscus is read, sinc(" the
color of the permanganate solution renders illlpO,~sible accllrate
readings of the lowest point.
The reactions involved arc as follows:

5Na 20204 + 8H 2S0 4 + 2KIVInOc~5Na~S04 + 10CO z + K2S0,~

5(133.99) 2(158.03)
+ 2MnSO.j 8H~O. +
50 20 4- - + 2Mn04- + 16H+---).10C0 2 + 2Mn++ + 8H zO.
5 X 2(-) + 2(-) + 16(+) = zero + 2 X 2(+)
Mn04-~Mn++ giving up 5( +)
0 2 0 4---200 2 taking on 2( +)
Therefore 2 moles of 1\1n04- are equivalent to 5 moles of 0 20 4----,
and ZKMn04 is equivalent to 5ND. zC ZO.1
The amount of sodium oxalate cquivl1hellt. to 1,000 CC. of 0.1
. . 5 X 133.99
N KM f n 0 4 ca IcuIat e d f rom,th e [t)OVl'
1 equatWJl IS 5 X 2 X 10
= 6.7000 Gm. Therefore 1 ce. of 0.1 N KMn()4 is equivalent
to 6.7000/1,000 = 0.006700 Gm. Na ZC Z0 4 If a 0.25 Gm.
sample of sodium oxalate required 34.20 CC. of 0.1 N KMn04
for titration, the normality of the l{MnO.j solution \-v<mld be
34.2 ~'~~00670 X 0.1 = 0.10907, and the solution should be
labeled 0.10907 N KMnO.j.
When permanganate solutions are diluted to an exact normal-
ity, water freshly distilled from permanganate solution should be
used to avoid the introduction of organip matter.
Potassiulll perlllanganate solution prepared as directed above
will retain its value for a long time if it is protected from sunlight
and from the entry of organic matter. It iioJ best, however, to
l'estandardize the solution from time to time. Solution of
standard potassium permanganate withdrawn from the stock
bottle should never be returned to it. Solutions of oxidizing
agents should never be titrated from Mohr burettes, since the
Qxidizing agent attacks the rubber and the solutions are thereby
weakened in concentration. The burette used to measure the
136 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

permanganate solution should be washed out immediately

after use to prevent the formation of a deposit of manganese
dioxide on the burette wall.
Questions and Problems
1. Define the following terms in accordance with the elect,ron theory:
(a) oxidation, (b) reduction, (e) electron, (d) unit positive charge.
2. How much KMn04 is needed to prepare 500 ce. of 2N, 0.1200 N,
0.02 N, and 0.01 N KMnO, solutions, respectively?
3. Why is the titration of permanganate with a reducing agent carried
out in acid solution?
4. Why should Mohr burettes never be employed to measure permanga-
nate solution?
DIRECT TITRATION METHODS
Exercise 38
To Determine the Purity of Ferrous Sulfate.-Potassium
permanganate is especially well suited to the estimation of iron
and ferrous salts oxidimetrically. When the titration is carried
out in solution acidulated with sulfuric acid, the ferrous ion
is oxidized quantitatively to ferric ion.
Object.-Assay of Ferrous Sulfate.
Materials Required.-About 1 Gm. of ferrous sulfate.
25 ce. of dilutecl sulfuric acid.
0.1 N potassium perlllanganate.
Procedure.-" Dissolve about 1 Gm. of Ferrous Sulfate, aceurately
weighed, in 25 ceo of diluted sulfuric acid, and titrate with tenth-normlll
potassium permanganate until a permanent pink color is produced. Each
ee. of tenth-normal potassium perrnanganate corresponds to 0.01519 Gm.
of FeS04."
After dissolving the sample in 25 cc. of diluted sulfuric acid,
it is best to wash down the sides and neck of the flask with about
25 cC. of cold distilled water. The titration must be conducted
in acid solution to maintain a sensibly constant hydrogen ion
concentration as explained on page 134. The pink color of
permallganate is distinct when a drop in excess of that required
to oxidize all of the ferrous salt to the ferric state has been added.
The reaction is as follows:
10FeS04 + 2KMn04 + 8H2S04~5Fe2(S04)3 + 2MnS04
10(151.9) 2(158.03)
 OXIDA'l'ION-REDUC1'/O}/ METliODS' 131

or, written ~onically,

10Fe++ + 2lVIl10 4- + 16H+---410Fe+++ + 2Ivln++ + 8H 20

In the above equations the water of crystallization is omitted,
since the U.S.P. assay determines the content of FeS04.
316.06 Gm. of KMn04 is equivalent to 1,519 Gm. of FeS04
and each cubic centimeter of 0.1 N i(Mn04 is equivalent to
1,519
100 X 1,000 = 0.01519 Gm. FeS04. The 100 ill the denominator
is obtained by taking the change of charge of 2KMn04 ",,10
multiplied by 10, the dilution of a 0.1 N ,mlution.
If the per cent of iron in the sulfate is desired, it can be cal-
culated by substituting its equivalent, 55.84, for that of FeS04.
Thus each cubic centimeter of 0.1 N KMn04 is equivalent to
10 X 55.84
100 X 1,000 = 0.005584 Gm. Fe. By the same procedure, the
equivalent of FeS04.7H 2 0 can be calculated.
Calculate the per cent Fe, FeS04, and FeS04.7H 20 equivalent
to the sample assayed. The U.S.P. requires that ferrOllR sulfate
contain 55.36 to 57.07 per cent FeSCh Compare YOul' results
with the official requirement.
Questions and Problems
1. Why is it Ullnecessary to cluploy 1m illdientor in direct titmtiollH with
KMnO,?
2. What quant.ity of electricity undergoes tl'!1l18fer in the oxidation of
two moles of FeSO. to Fe2(SOl);'?
3. Does the change of charge on' the iron in question 2 corn~spond with
the change in valence?
4. What must be the normality of a solution of KMnO. in order that each
ell bie cell timeter shall be cq ui valen t to 0.1 per cent FeSO.?
5. How much FeSO. would be required to prepare 500 ce. of 0.1 N solution
of FeSO,.7H.O?
6. What other official ferrous salts are assayed by methods analogous to
tlutt given above?
Exercise 39
To Determine the Purity of Reduced Iron.-In this assay the
metallic iron is converted quantitatively into ferrous ion, and
estimated oxidimetrically by titration with potassium permanga-
nate solution
138 QUAN1'jTATIVE PHARMACEUTICA.L CHEMISTRY

Object.-Assay of Reduced Iron.

Materials Required.-About 1 Grn. of reduced iron.
10 Gm. of nwrcuric chloride.
20 cc. of diluted Hulfuric acid.
l.0 N potassium pcrlllangHIlate.
Procedure.-l. "Triturate thoroughly about 1 Gm. of Reduced Iron,
weigh accurately, plnee it in a 100 ce. volumetric flask, ltnd add 10 Gm. of
fillety powdered corrosive mercuric chloride and 50 cc. of boiling distilled
\vnter."

The rcdueed iron is triturated to produce a uniform mixture

in order that a representative sample may be obtained for assay.
Since reduced iron always contains some oxide, the metallic
iron in the Hample is oxidized by boiling it in solution with mer-
curic chloride. The mereurie ehloricle oxidizes the metallic
iron to the ferrous state without changing any oxide present.
HgCb + Fe-->FeCb + Hg
or, written ionically,
Hg++ + Fe--> Fe++ + Hg
2. "Heat the mixture for ten minutes on a water bath, shaking it fre-
quently, thon fill the flask to the 100 co. mark with distilled water, recently
hoiled and cooled, and cool the mixture to room temperature. Again fill
the flfl,sk to the mark with distilled water, stopper, agitate the contents well,
and nUow to stand for a few minutel:l."
The solution is diluted to exactly 100 cc. so that an aliquot
portion representing oue-fifth of the weight of sample (20 cc.
of solution) may be obtained.
3. "Now filter the contcntl:l of tho flask, and immediately titrate 20 CC.
of the filtrate to which has been added 20 cc. of diluted sulfuric acid, with
tenth-norm[,l potassiUlll permangmutte, until a permanent pink color is
produced. Each cc. of tenth-normal potassium permanganate is equivalent
to 0.005584 GIll. of Fe."
The aliquot portion is filtered to free it of any finely divided
mercury or iron oxide which may be suspended in the solution.
The titration is conducted in acidulated solution in a manner
similar to that described in the preceding assay.
The reactions involved in the titration and the calculations
are also similar. Write all reactions and calculate the purity
of the sample assayed.
 OXIDATION-REDUCTION METHODS 130

Questions and Problems

1. Does the iron gain or lose electroTlS wh()fi oxidizer! to the ferron!'; state
by HgCI.? What quantity of electrieit;,{ dot's it gain or lo"e pOl' moho of Fe?
2. Given pure iron, how might a solution of potassiulll pernwngalwte he
stand!mlized against it?
3. If the iron in a 0.15 Grn. sample of iI'OIl ore rccillircs 15.0 ceo of 0.1 N
KMnO. for oxidation from FeN to Fe 1-++, what is the percentage of Fe ill
the are?

Exercise 40

Object.-Assay of Solution of Hydrogen Peroxide.

Materials Required.-Ahout 2 ce. of l:iolntioIl of hydrogen pemxide.
20 cC. of diluted sulfuric !wid.
0.1 N potassium permangallltte.
Procedure.-"Measure aecllrately 2 ce. of Solution of Hy,lrogl'll PNoxic1e
and transfer it to a suitable flask containing 20 m!. of distiller! wnter. Acid
20 ce. of diluted s~llfuric twirl, and titrat(~ wil 11 tcmth-Ilormn! pot a,"illlll
perml1nganate. Each eubic cClltimrtcr of tenth-normal puta;;.<iulJl prr-
manganate is equivalent to 0.001701 Gill. of H 2 0,."

The reasons for the various steps ill the above nssay htwp
already peen given under the assay of ferrous .~ulf!1te. Thi:'l
assay illustrates a case where an oxidizing agent, H20~, serves
as a reducing agent for KMu04 and is itself reduced. ""lwu
H 20 2 acts as a reducing agent, 1 Gm.-atom of OXygC'1l from eueh
mole of H 20 2 is capable of uniting with 1 Gm.-atolll of oxygen
from the substance rcdu(;ed (KMnO.l) to form 1 mole of oxygen
gas:

2HO +0 (from oxidizing agcnt)----)H 20 +O 2

In the assay reaction 5 Gm.-atoms of oxygen from 5 moles of

H 20 2 unite with 5 Gm.-atmns of oxygen from 2 moles KMn04
to f~rm 5 mol~s of oxygen gas:

5H 20 2 + 2KMn04 + 3H2S0r-~K2S04 + 2j.\ilnS04 + 8H 0 2

5(34.02) 2(158.03) + 50 2
Each cubic centimeter of 0.1 N K1V111 0 4 is equivalent to
5 X 34.02
100 X 1,000 = 0.001701 Om. H202
140 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

To comply with the U.S.P. definition, solution of hydrogen

peroxide should contain not less than 2.5 and not more than 3.5
per cent of H 20 2 Calculate the percentage of H 20 2 in the sample
assayed and compare the strength of the solution with the U.S.P.
requirement.
Questions and Problems
1. A 2 cc. sample of hydrogen peroxide rcquired 8.5 ce. of KMnO. solution
for titration. If each cubic centimeter of ICMn04 is known to be equivalent
to 0.007294. GIll. Fe, what percentage of H.O. docs the sample contain?
2. Is H 2S0 4 necessaTy to supply a concentration of H+ to combine with
the oxygen liberated from the perlllanganute to form water? Why is the
H.SO. necessary?
3. Sodium perborate when dissolved in diluted sulfuric acid solution yields
NaHSO. + HaBO. + H 20.. Ascertain the method of assay of sodium
perbol'ate, balance the reaction involved in the assay, and compare the
method with thltt employee! in the :tssay of hydrogen peroxide.
4. Could lcud dixoide be assayed in this way? (See U.S.P. Teagent
PbO . )

TABLE XIV.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT Tl'J'RA'l'ION WITH

0.1 N KMn04

E1IUivl1-
Amount
lent of
used, Official requirement,
Substance 1 ce.
Gm. or per cent
0.1 N
ce.
KMnO,

U.S,P.
Ferrons ~u1fate .......... 1 0.01519 FeSO, = 54..36 to 57.07
Hydrogen peroxide solu-
tion .................. 2 0.001701 H 20 2 = 2.5 to 3.5
Hydroxylamine hydro-
chloride, R ........... 0.1 0.0034.75 NH 20H.HCl = 95
Indigo carmine, R, , , .... 0.3 0.01333 C IG H sO.N2(SOaNa)2 = 82
Iron, reduced ........... 1 0.005584. Fe = 90
Lead peroxide, R ........ 0.5 0.01196 PbO. = 90
Sodium perbomt.c ........ 0.25 0.0008 Available O2 = 9
Sodium peroxide, R ...... 0.7 0.0039 NI1.02 = 90
.-
R. "" reagent.
 OXlD.j r['ION-RIWUC'l'J ON M E'l'HODS 141
INDIRECT TITRATION METH.ODS
Exercise 41
Object.-Assay of Calcium Glueonate.
Materials Required.-0.5 Gm. of calcium glue onate.
2 ce. of hYflroehloric acid.
Ammoniulll oXlllnJe, T.S. (3.5 Gm. lLmmonium oxalate ill 100 ee. of wMer).
Dilute sulfuric aeid (1 iu 3).
0.1 N pot!tssium pcrmangmwtc.
Procedure.-l. "Weigh aecl1l'utely about 0.5 Gm. of Calcium Ghwonate,
dried to comJt.ant weight in a desiccator over Rlllfurie add, pl!tce it in a
250-cc. heaker, add 2 ce. of hydrochlorie ILcid anti 100 ceo of distilled water.
Heat the mixture to boiling, make the solution alkaline with ammonia T.S.
l>nd add, with ;.;Lining, an exct'sl'l of hot ammonium oxalate T.S. I-Ie!Lt the
mixture on a water bath during onc hour, filter through h[1t'dened filter paper
and wash thoroughly with warm distilled w[tter."

The caleium gluconato reacts with the HOI as follows:

CH 2 0H(CHOH)4.COO zCa.H 20 + HCI 2CHzOH(CHOH)4.
COOH + CaCl 2 + H 20
448.27
When ammoniuIll oxaln.te is added to the reaction mixture, the
calcium iOll is quantitatively precipitated as oxalate and the
above retwtion is forced to the right,
CaCh + (NH.l)2020c~CaC204 + 2NH4CI
The precipitation is earried out in hot solution and the mixture
is digested to secure a coarse grained precipitate.
2. "Pullcture the filter paper, wnsh the precipitute into a beaker by
means of a stream of hot distilled wHter, followed by 30 cc. of dilute (1 in 3)
sulfuric acid. Heat tho solutioJl, if necessary, to 60C. and titrate with
tenth-Ilormal p()ta~si\l!l1 pormnllganaie. Each cubic centimeter of tenth-
normal potaRsiulll perlIlllllgallate iH equivalent to 0.002804 Gm. of CaD."
The precipitate is washed with warm water to 'remove the
excess ammonium oxalate. Upon the addition of diluted sulfuric
acid, the calcium is precipitated as sulfate leaving; free oxalic
acid in solution,
Ca0204 + H2S0c-~CaSO1 + (COOHh
142 QUA. NTI1'ATIVE PHARMACEU'l'ICAL CHEMISTRY

and the amount of oxalic acid liberated is found by titration with

standard KMl10 4 solution. The reaction indicated proceeds to
completion because the oxalic acid formed is consumed by titra-
tion with the permanganate.
This is an indirect method since neither the calcium nor the
gluconic acid is determined directly but the calcium is determined
by the indirect process of estimating the amount of oxalic acid
with which it combined.
Calculate the per cent of calcium gluconate in the sample
assayed.
Questions and Problems
1. Calcium glucona.te is required to yield not less than 12.4 and not more
than 12.8 per cent, of CaO. Within what limits of per cent purity would
calcium glllc;oUl1te conform to these requirements?
2. Wh:Lt amounls of anhydrous calcium lactate might 5 grain tablets or
calcium lactate contain and meet the requirements of tbe N. F.?
TABLE XV.-Oli't.'ICIAL SUBSTANCES ASSAYED BY PRECIPITA'l'ION WITH
AMMONIUM OXALA'rE AND TITRATION OF THE OXALIC ACID FROM
THE PRECIPITATE WITH 0.1 N KMn04

Equiva-
Amount
lent 01 Ollicin!
used,
Substance 1 cc. requirement,
Gm. or
0.1 N per cent
cc.
KMnO.

U.S.P.
Calcium bromide ....... 0.4 0.009996 C"Br2 = 84 to 94
Calcium gluconate ......... , 0.5 0.002804 C"O = 12.4 to 12.8
C"lcium lactate ............. 0.5 0.0101)1 (CH3CH.OH.COO),Co. = 98
N.F.
Calcium carbonate, tablets of. 0.0 0.005004 c.. eo, = 92,5 to 107.5b
Calcium glucollute, ampule of. O.5a 0.02242 C12II"0,,C:)'.H,O = 05 to 105 b
C"lcium laotale, tablets of .... 1. 5a 0.01541 (CH,ClI.OH. COO) ,Cn.5H20 = 92,5
to 107.5 b

a Weight of ingredients sought.

b Per cent of the labeled amount.

RESIDUAL TITRATION METHODS

The residual titrations in the official assays with potassium
permanganate solution are of three types, namely: (1) An excess
of standard potassium permanganate solution is employed to
oxidize a substance, and the amount in excess is determined by
 OXIDATION-REDUCTION METHODS 143
residual titration with standard oxalic aeid solution; (2) an excess
of standard oxalic acid solution is added to a substance and the
excess of oxalic acid is titrated with standard permanganate
solution; (3) an excess of ferrous sulfate solution is added to an
oxidizing agent, and the amount of ferrous sulfate ill excess is
titrated \vith standard pennanganate solution.
The following exercise is an example of a residual titration
method in which an excess of standard permanganate solution
is employed to oxidize a substance, and the :1111011nt in excess is
determined by residual titration of standard oxalie acid solution.
Exercise 42
Object.-To Prepare and Standardize n.1 N OXlllie Add.
Materials Required.-6A5 Gm. of reagent oxalic acid.
0.1 N potassiam perlllI1nglilla teo
Procedure.-Dissolye 6.45 Gm. of reagent oxalic Ilcid in Rufficicnt dis~
tilled water to mel,snre 1,000 ceo AscerLain its cxaet strfl!)!!:th by titration
against freshly stltfl(lardized 0.1 N potassiulll permangallate [tIll[ adjust to
IJxacv normality or to a known strength of llpproximate normality.

The purest oxalic. add obtainable varies in composition due to

differences in its moisture content. If the acid is heated suffi-
ciently to drive off absorbed moisture, it tends to lose water of
crystallization. Consequently, a solution of approximate con-
centration is prepared, and its nonnnJity is determined by
titration against a standard solution, in this case, 0.1 N KMn04.
The titration is conducted a,s follows:
Measure accmrately from a burette about 30 CC. of the acid
solution into a beaker and dilute with about 200 cC. of distilled
water. Add 7 cc. of sulfuric ac.id, heat to about 70C. and then
slowly add the standard permanganate solution from a glass-
stoppered burette, with constant stirring, until a pale pink color
is produced which persists for 15 sec. The temperature at the
conclusion of the titration should not be less than 60C. Calcu-
late the normality of the solution and, if desil'ccl, adjust the
solution to exactly 0.1 lv. The calculations and reactions
are similar to those described in the preparation of 0.1 N KMn04.
The probability of error is minimized by standardizing the
oxalic acid solution against the same permangunate solution
with which it is to be used in analyses. The normality of ar~
144 QU.1NTIT:1TIVl!} PIL1RivIACBUTICAL CHEMISTRY

oxalic acid solution may be determined by titration with l-3tundard

:Llkali as follows:
Dilute 25 ce. of 0.1 N N aOR aecurately measured from a
burette with 25 cc. of distilled water, add 2 to 4 drops of phenol-
phthalein T.S. as indicator, and heat the solution to boiling.
Titrate this solution with 0.1 N oxalic acid unW the pink color
disappears. The solution should be brought to a gentle boil at
the end to expel all CO 2 which, if present, would affect the
phenol ph thalein indicator.
The oxalic acid is dibasic; that is, one molecule requires two
of sodium hydroxide for neutralization. Since 1 ee. of 0.1 N
acid is equivalent to 1 cc. of 0.1 N alkali, the normality of this
solution may be calculated eaRily: .
Cc. alkali X N . .
C .1 = normalIty of aCId
c. aOIC

Oxalic aciel solution deteriorates on standing and must be

restandardized from time to time.

Questions and Problems

1. Why is hot titration employed?
2. What is the cquivalenL weightof oXlllie twid a;,; an acid; as a reducing
agent?
3. Write the reaction involved in the standardization of oxalic acid
against sodium hydroxide solution.
4. How much oxalic acid is required to prepare 1,000 ce. of solutions of the
following respective normalities: (al 0.5000, (Ii) 0.2500, (clO.200?

Exercise 43
Object.-Assay of Sodium Nitrite.
Materials Required.-1 Grn. of sodium nitrite.
50 cc. of 0.1 N potassium permanganate.
5 cc. of sulfuric acid.
0.1 N oxalic acid.
Procedure.-l. "Dry about 1 Gni. of Sodium Nitrite to constant weight
over sulfuric acid, weigh accurately in a stoppered weighing-bottle, clis-
~olve the salt in a volumetric flask with sufficient distilled water to make
ce.
100 cc., and add 10 ce. of this solution, from It pipette, to a mixture of 50
oi tenth-normal potassium permangallate, 100 ce. of distilled water, and 5
cc. of sulfuric acid. When adding the sodium nitrite solution, immerse the
tip of the pipette beneath the surface. of the permanganate mixture."
 OXIDL1TION-RIWU(!'!'WN ME'!'HODS 145

Sodium nitrite is deliqnescent lind is therefure directed to he

dried to constant weight in a clci"iccator oYer Bulfuric aeid
preyiotls to weighing. BeCllUf:ie of thi~ prol)erty, the f;umple j~
also directed to be weighed in a stoppered 'n~ighing hottle.
Nitrous aeid is oxidized by the permangal1nte to uih'i(' acid.
If the Holution of sodium nitrite we're acidulated with sulfuric
acid, nitrous add, which is yolatile, would be lost.. The sodium
nitrite is therefore introduced beneath the surface of the acidu-
lated perll1anganate solution.
2. "Warm the liquid to 40C., allow it, to stand for five minutes, and add
25 ce. of tenth-normal o~alic ncid. Heat the Jlli~tnre to n.hout SOC., and
titrate with tenth-normal potassiulll penn::mgalJute. Each cuhill l1ent.imett'l'
of tcnth-normal potassiulll perIllangaunte io equivalent to 0.003,150 Gill. of
NaN0 2."
The oxiclation of nitrou~ to nit.ric acid proceeds slcHvly at
ordinary temperature, but it is rapid at 40C. The warm mixture
is allowed. to stand for 5 min. to imure completion of the oxida-
tion. The excess 0.1 }loxtl,Ue ttcid i8 then determined by titra-
tion with permanganate until the pink permanganate color if!
obtained.
The real'tioll may be represented as taking place in two st.ages,
viz.:
2NaN0 2 + H2S0c--~Na2S0.1 + 2HN0 2
2(69.01)
.5HN0 2 + 2KMn04 + 3H 2SOc-+.5HNOa + 1(2S04
+ 2MnS04 + 3H 0 2

In this reaction each nitrogen atom loses two electrons and

undergoes a change of polarity of two. Tlwl'Cfore, 1 ec. of 0.1
N KMn04 is pqllivulent. to 2" X 1~9~\,ooo = O.(l03450 Gm.
NaN0 2 Calculate the purity of the sample assayed.
. Questions and Problems
1. Why should the tip of the pipette he immersed beneath the surface 0\
the pel'manganate solution when the solution of iSodhim nitrite is added?
2. Why is the reaction mixtttre wurlllPd and allowed to stand?
3. Write ionic ally the l'oactions involved in the above assay.
4. How Illueh 0.0985 N KMl10 4 would be required to titrate 0.8544 Gm.
of NaN02 if the NaN0 2 is 92 per cent, pure?
5. Could nitrous acid and other nitrites be astiayed hy the ahove method?
146 QUANTITATIVE PHARM J1CEUTICAL CHEMISTRY

Sodium nitrite and potassium nitrite, reagent, are *the only

official substances assayed by this method.
The following exercise illustrates residual titration methods in
which an excess of standard uxalic acid solution is employed to
precipit!1te a substance quantitatively with subsequent residual
titration of the excess oxalic add by standard pel'manganate
solution.

Exercise 44
To Determine the Purity of Calcium Carbonate.-Caleium
and lead salts can be assayed volumetrically by precipitation
with standard oxalic acid added in excess and subsequent titra-
tion of the excess acid with standard potassium permanganate
solution.
Object.-Assay of Precipitated Calcium Carbonate.
Materials Required.-About 0.4 Gm. of precipitated calcium earbonate.
10 cc. of diluted hydrochloric acid.
100 cc. of 0.1 N oxalic acid.
Ammonia water, 10 per cent.
Diluted sulfuric acid.
0.1 N potaH~ium permanganate.
Procedure.-l. "Dry about 0.4 Gm. of Precipitatod Calcium Carbonate
to constant weight at 200C., weigh accurately, dissolve it in a mixture of
10 cc. of diluted hydrochloric acid and 10 ce. of distilled water, and boil the
solution to expel all carbon dioxide."

Calcium carbonate is soluble in hydrochloric acid because the

product of the concentration of calcium and carbonate ions
becomes less than the solubility product of calcium carbonate.
When a strong acid is added to calcium carbonate, the concen-
tration of COr- ion is decreased, due to the formation of slightly
ionized carbonic acid:

CaCO g Ca++ + C0 3- -

C0 3- - + 2H+ H 2CO a CO 2 +H 02

The increase in H+ concentration displaces the equilibria

toward the right and leads to solution of the calcium carbonate
in hydrochloric acid. 'rhe CO 2 formed in the reaction is expelled
by boiling.
 OXIDATION-REDUCTION j~fET[JODS 147
2. "Transfer this solution to tL 200 ee. volumetric fbsk, add 100 ce. of
tenth-noDllal oxalic acid, render it alkaline with ammonia T.S., shake the
mixture well, and allow it to stand for three hours at from 00 0 tn 70C., or
over night at room t.emperature. Cool the liquid if necesoary, fill the Rusk
with distilled water to 200 ce., mix weU, filter through a dry filter into !l.
dry flask, and reject the first 20 ce. of the filtrate."

The 0.1 N oxalic acid is added to the solution contained in the

200 cc. volumetric flask and thc whole iR made alkaline with
ammonia T.S. added slowly with continuous stirring. Ammo-
nium oxalate is formed, and it reaets with the calcium chloride
to form ammonium chloride and insoluble calcium oxal::Lh~. The
exceRS of ammonium oxalate present decreases the solubility of
calcium oxalate to a negligible quantity by common ion effect.
The Rolution is digested at 60 to 70C. to prolIlotfJ the growth of
larger crystals and to permit ('.omplete precipitation of the
calcium oxalate. The solution is eoolc~cl, Rime ealeiml1 oxalate is
more soluble in hot than in cold water, its solubility being 5.6
mg. per liter at 20C. and 14 mg. at 95C. The solution is
made up to a precise volume, 200 ce., and an aliquot portion,
100 cc., is taken for residual titration. ThiR procedure eliminates
the tedious washing whieh would he required to render the erys-
talline calcium oxalate free of ammonium oxalate. The filtm-
tion of the aliquot portion is best conducted by collect,illg the first
20 ce. of filtrate, which is to be rejected, in a cylindrical graduate,
then transferring the funllel to a 100 ce. volumetric flask and
filtering an additional quantity of solution sufficient to fill it
exactly to the graduation mark.
3. "Acidulate 100 ceo of the filtrate (corresponding to one-half of the
Precipitated Calcium Carbonate taken) '''ith diluted sulfuric acid, then
add 25 ceo more of the diluted sulfuric add, warlIl the solution to about
GOC" and titrate with tenth-normal potassium pernulnganate. Each
cubic centimeter of tenth-normal oxalic acid is equivalent to 0.005004 GIll. of
CaCO a."
Thc filtrate is transferred quantitatively to a 250 ce. Erlen~
meyer flask, acidulated with sulfuric acid and titrated with 0.1
N KMn04 to determine the excess oxalic acid.
The reactions involved are:
1. Solution of CaCO a in HCI:
CaCO a + 2HCl---+CaC1 2 -t- CO 2 +H0 2
148 QUilNTl'l'ATlVE PHARMM,'EUTICAL CHEMISTRY

2. Addition of NH 40H to H 2 C 20.!:

H 2C 20 4 + 2NH 40H->(NH 4)2C 2 0.! + 2H 20
3. Double decomposition between CaCl 2 and (NH 4) 2C Z0 4:
CaCl z + (NH4)2C 20 c ---+CaC 20 4 + 2NH 4CI
4. Filtrate of (NH4)2C204 acidulated with H 2S0 4 :
(NH4) ZC 20 4 + H 2S0 ---7H C 0.! + (NH )2S0'J
4 2 2 4

5. Residual titration of excess H 2C 20 1 with KMn04:

5H 2C 20. + 2KlVInQ'1 + 3H SO. -)I\: S04 + 2lVInS04
2 1 2

+ 10CO z + 8H zO
The calculation of the purity of the sample may be made
according to the following:
One cubic centimeter of 0.1 N oxalic acid iR equivalent to

2 X ~~0~81,OOO = 0.005004 Om. CttCO a

[eN x CCL H 2C 204) - (N X cc. KMnO.l X 2)] X 0.005004
Wt. of sample
X 10 X 100 = per cent purity
Note that the number of cubic centimeters of KlVIn04 solution
is multiplied by two, since only one-half of the excess oxalic acid
is represented in the aliquot portioll used in the residual titration.

Questions and Problems

1. Why is the solutioll rendered alkaline wit.h NH 40H?
2. How does common ion effeet serve to make the precipitl1tjOll of calcium
oxalate complete?
3. Why is the lllixtlll'e digest(~d at 60 to 7000.?
4. Why is the soluUon cooled previous to dilution and filtration?
6. N~lme I-lcveral official calcium compounds whieh arc assayed by methods
uualogous to that given above.
6. Look up the oH1cia,l method of aRH~lY for solution of lead subaeetate,
Explain each step i1) the pl'Ocedure Hnd write all reactions.
7. An unknown calcium salt weighillJ.!; 0.25 Gm. assayed by the above
method was treated with 100 ce. of 0.1010 N oxalic acid. Upon residual
titration of an aliquot portion representing one-half of the excess oxalic
acid, 22.45 ce. of 0.0982 N KMnO. wa~ required. To what percentage of
Ca, CaO, OaOOa, and CaOh, respectively, is the calcium sa.lt equivalent?
 OXIDilTION-RBDUCTION ilfE7'lIO[)S

TABLE XVI.-OFFICIAL SUBSTANCES ASSAYBD BY PnECIl'ITATION WITH

EXCESS STANDARD OXALIC ACID SOLUTION AND RESIDUAL TITIlATl()N
OF TIlE EXCES;; OXALIC ACID WITH 0.1 N KMnO 1

Amonnt
used,
Equivil- i
lent of Ii
Oflicial
Substatwc TI~qllireIllcnt,
Gm. ce. 0.1 NI per cent
or ce. H,C,O. I
___ - __J --_________
U.S.P.
Calcium carhonate, pl'P.cipitILte(1
Calcium chloride. . . . . . . . . . . . . .
0.4
II .3
(). 005(}0.J C:lCO~= 98
0.00555 C'1.Cb = 75 to f>5
Calcium chloride, fused, R.. . . . . 0.3 O. 00555 CaCl~ = f}4
Calcium hydroxide. . . . . . . . . . . . . 0 .3 0.003705 Ca(OB)2 = 1).5
Chalk, prepared. . . . . . . . . . . . . . . 3.5 () . 005004 CI1.CO~ = 97
Lead acetate. . . . . . . . . . . . . . . . . . 5 0.01()20 I Pb(C 2H:102h ""
85.31 to SO.57
Lead monoxide, R. . . . . . . . . . . . . 0.4 0.01110 PbO=98
Lime, R....... .......... ..... 1 0.002804 CaO = 95
N.F.
Caleilllu chloride, ampuls of. . . . . 0.25" 0.007351' CaCI . 2II 20 "" 95
t.o 105b
Lead monoxide. . . . . . . . . . . . . . . . 0 .4 O.OIllO PbO = 97
Lead subacctate, soilltioll. . . . . . 1 0.01036 Ph = 22.5W/V
Lead subacetatc, solutiun, clilu tp 50 0.01030 Pb = 0.7 to
O.81Y1V
Lime (calcium oxicle) .......... . 0.002804 CltO = 95
\
R. = reagent..

n Amount of ingredient Bought .

Per cent of the labeled amount.

Potassium permanganate, U.S.P., and mangaJleSe dioxide and

tablets of potassium permanganate of the N.F. are assayed by
treating the sample 'with an excess of standard oxalic acid and
titrating the excess uf add with stunclu.rd permanganate.
The following assay illustrates a method involving reduction of
fLU oxidizing agent by means of an excess of ferrous sulfate, and
residual titration of the excess ferrons sulfate (unuxidized)
by Rtandard permanganate solution:
Exercise 46
Object.-Assay of Potassium Chlorate.
Materials Required.-Q,l Gm. of pot.assium chlorate.
150 QUANTITA'lTVE PHARMACEUTICAL CHEMISTRY

100 cc. of acid felTous sulfate solution prepared by dissolving 7 Gm. of

ferrous sulfate erystals (FeS04' 7I-I 20) in 90 cc. of freshly boiled distilled
water and adding sufficient sulfuric acid to make 100 CC.
20 CC. of Imtuganous sulfate solution prepared by dissolving 11 Gm. of
manganese sulfate (MnSO .4H20) in 50 ce. of distilled water and adding
sufficient sulfuric acid to make 100 cc.
0.1 N potassium permanganate.
Procedure.-l, "Weigh accurately about 0.1 Gm. of Potassium Chlorate,
diHsolve it in 10 ce. of diHtilled water in !1 250-ec. flask, !1nd add, from a
burette, 35 cc. of acid ferrous sulfatfl T.S. Prepare a valve-stopper by tak-
ing a piece of rubber tuhing of convenient. diameter and about 5 em. in
length, place a piece of glass rod ill one. end, and slip the other end over a
glass tube whieh passes through a perforated stoppel' of a size to fit the
flask used. Cut. a longituc\ill!tl slit about 15 mm. long in one side of the ruh-
bel' tube about midway of its length. Insert this stoppel' in the flask and
boil the mixture for ten minutes."

When potassium chlorate is treated with ferrous sulfate T.S.

in acid solution, the ferrous salt is oxidized to the ferric state
and the potassium chlorate is reduced to potassium chloride:
KCIO a + 6FeSO'1 + 3H S0 ->J{CI + 3Fe2(S04)a + 3H 0
2 4 2
122.55
or
C1O'3- + 6Fe++ + 6H+ = Cl- + 6Fe+++
Sufficient acid must be present to supply an abundance of H+
to satisfy the equation and to prevent the hydrolysis of the ferric
sulfate. If oxygen from the ail' were allowed to enter, it would
oxidize some of the FeS04 to Fe2(S04)a. The reaction is, there-
fore, carried out in a flask fitted with a Bunsen valve which allows
steam to escape but prevents the entrance of air.
2. "Cool the mixture, lldd 10 ec. of mangllllous sulfate T.S., and titrate
the excess of ferrous sulfate with tenth-llormal potassium pel'mangnnate."

The mixture should be cooled in the flask protected from air

and the manganous sulfate solution added just before titration
of the residual ferrous sulfate with standard potassium per-
manganate solution. The presence of manganous sulfate is
essential in the reaction mixture. The manner in which it
functions is not clearly understood, but it is known that in the
presence of chlorides more permanganate is used up than is
necessary to oxidize the ferrous salt to the ferric condition. The
 OXIDATION-REDUCTION ME1'HODS 151

manganous iOllS react with perrnanganate iOlls to form man-

ganese ions Mn++++. In this state of oxidation, the mangallese
is unstable in acid solution, and it is reduced more reaclily by
ferrous ions than by chloride ions.
3. "Conduct lL blank experiment with another porticn of 35 (;e. of neil!
ferrous sulphate T.S., measllred from :1 hllmtte, and Huhtmd the result,
of the former titration from that of th(, latter. Each ellbie ('en tinwtnl" of
tenth-normal potassium pernumgarmte iH equivalellt to 0.00204:3 Gill. of
KCl0 3 ."

A blank experiment is conducted employing all of the reagents

used in the assay but without the addition of potassium chlorate.
This procedure is necessary, since the exact concentration of t}w
ferrous sulfate T.S. is not known. The difference l)(~hvef'n the
volume of 0.1 N Kl\'ln04 eOllslIUled in the blank experiment
and in the assay gives the number of cubic centimeters of 0.1
N KMn04 equivalent to the sample> of potassium chlorate.
Each cubic centimeter of 0.1 N KMn04 consnmed ill the
titration of the residual ferrous sulfate is equivalent to

6 X ~~2~51,OOO = 0.002043 Om. KCIO a

If 50 cc. of ferrous sulfate T.S. requires 49.5 cc. of 0.1 N

KMn04 in the blank experiment ~tnd the residual ferrous sulfate
in the assay requires 4.5 ec. of 0.1 N KMnO'I, the 0.1 Om. sample
of KOI0 3 is equivalent to 49.5 - 4.5 = 45 ce. of 0.1 N Kl\ifn04.
The per cent purity of the KOlOa may then be calculated:
0.002043 X 45
0.1 X 100 = 91.93 per cent

Questions and Problems

1. Why is the reaction carried out ill a flask fitted with a Bunsen valve?
2. Explain how the equivalent weight of leCIO. is derived Oil the hasis of
change of charge ..
3. Why is it necessary to rUll a blank exporiment?
Potassium chlorate and tablets of potassiuUl chlorate are the
only official substances assayed by this method.
Dichromate Methods.-In certain official preparations con-
taining ferrous salts the percentage of iron salt cannot be aceu~
152 QUAN1'I'l'A'1'IVllJ PJIARM.1CEUTICAL CHEMISTRY

rat ely estimated by oxidation in acidulated permanganate

solution owing to the presence of sugan; which l'PCluce this rea-
gent. Potassium dichromate possesses two distinct advantages
over potassiulll permanganate a8 'a volumetric reagent, namely, it
can be obtained very pure, and it does not deteriorate upon aging
if properly kept.
Determination of the End Point.-The end point is determined
in the official assays by the use of diphenylamine as the indicator.
In the presence of sugars, the violet color formed by this indicator
is obscured somewhat by the formation of the green chromic ion.
The end point of titration may also be determined by the use
of an "outside indicator" as follows: The end point of titration
is found by placing a drop or two of freshly prepared potassium
ferricyanide solution in a number of the depressions in a porcelaill
spot plate. A clrop of the solution being titrated is brought in
contact with this indicator by means of a stirring rod at frequent
intervals in the titration. As long as ferrous iron is present, a
blue coloration results when the solution comes in contact with
the indicator, the end point being reached when the blue color
is no longer produced. It is usually advisable to run a prelimi-
llary titration so that it will not be necessary to withdraw an
undue quantity of reaction solution during the titration.
The following reaction takes place between ferrous sulfate
and potassium ferrieyanide:

3Fe80 4 + 2K3Fe(CN)u~Fe3[Fe(CN)6h + 3K 80 2 4
blue

Exercise 46

Object.-Preparation of 0.1 N Potassium Dichromate.

Materials Required.-1.2 Gm. of pure potassium dichromate.
Procedure.-Pulverir.e about 1.2 GIll. of pure potassium dichromate and
dry the product to constant weight }1t. 120C. Accurately weigh exactly
0.9807 Gm. of dried K 2Cr,07 and dissolve it ill sufficient distilled water to
make 200 cc. at the temperature at which Uw volumetric flask has been
calibrated. If the dichromate used is pure, t.he solution will he exactly
tenth~norm.al.

Each chromium atom in K 2 Cr 2 07 has a polarity of 6( +) and

upon reduction two trivalent chromic ions are formed. Each
 OX!DA1'lON-REDUC'l'/ON METHODS 153

chromium atom undergoes a ehange in polarity of three. Since

two atoms of chromium undergo the change, the total change in
polarity iR 2 X 3 = 6. A normal solution of K2Cr207 should,
therefore, contain one-sixth of a mole per 1,000 cc.
An approximately 0.1 N solutioll of K2Cr207 can be standard-
ized against pure iron wire or 0.1 N sodium thiosulfate solution
(sec page 158).

E:ocercise 47

Object.-Assay of lVIns:,; of Ferrous Carbonate.

Materials Required.-Ahout 1 Gm. of mass of fcrl'ou~ carhonate.
15 ce. of diluted sulfurie add.
0.1 N potassiuUl diehromate.
Diphenylamine, T.S. (1.0 Gm. of diphenylamine in 100 co. of nmgfmt
H2S0.).
Procedure.-" Dissoh'c about 1 Glll. of Mass of }ferrous Cllrbonat(),
ttecl1rately weighed, in 15 ce. of diluted sulfuric !tcid, dilute the solution wit.h
distilled water to about 150 (!c., and immediately tHmte with tenth-normal
potassium diehromntr, using about 0.5 cc. of diphenylamine T.S. as the
indicator. Eaeh ~llhie centimeter of tenth-normal potaosium dichromate is
equivalent to 0.01158 Gtn. of FeeO,."

The mass of ferrous carbonate difisolves in dilutccl Rulfurie

acid to form ferrous sulfate, /liz.:

Sufficient acid in excess of that required for the formation of

ferrous sulfate is needed to supply a constant source of hydrogen
ion:

The solution is diluted to facilitate the reaction in the presence

of the suspended matter and to minimize the error caused by
the formation of the green chromic ion which tends to obscure the
violet color of the diphenylamine at the end point. The titration
should be conducted as SOon as possible after solutIon of the
sample has been effected to avoid as much as possible the oxida-
tion of ferrous to ferric iron by the action of atmosphelic oxygen.
154 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The reactions which occur in the assay are:

FeCO s + H2S0.1~FcSO'1 + H 20 +'C0 2
115.84
61<'e80 4 + + 7H280r~3Fe2(SO'I).1 + K
R 2Cr 2 07 2 S0 4
+ Cr 2(80 4)8 + 7H zO
or

Each Fe++ undergoes a change of charge of one. Each cubic

centimeter of 0.1 N 1(2Cr 207 consumed in the titration is there-
fore equivalent to 10 1~51~~OO = 0.01158 Gm. FeC03. Cal-
culate the per cent of iron and the per cent of ferrons carbonate
in the sample assayecl. The U.S.P. requires tlutt mass or ferrous
carbonate eontain 36 to 41 per cent of FeCO a Compare the
per cent of ferrous carbonate found in the assayed sample with
the official requirement.
Questions and Problems
1. Why mnst the solution be ac:i<l in reaction dming titration?
2. If three pills of ferrous carbonate of equal weight employed as a sample
require 16.25 ce. of 0.1 N K,Cr,07, what amount of Fe and of FeCO a,
respectively, doe~ each pill contain?

TABLE XVII.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH

0.1 N K,CI' 207

Equiva- Official
Substanee Amount
lent of requirement,
used
1 cc. per cent
._-----
U.S.P.
Mass of ferrou8 c!1Tbonate ......... 1 Gm. 0.01158 FeCO a = 36 to
41
Pill of ferrous carbonate .......... 3 pills 0.01158 FeCO a = 0.06
Gm. per pill
N.F.
Saccharated ferrous carbonate ...... 2 Gm. 0.01158 FeCO a = 15
 CHAPTER IX
OXIDATION AND REDUCTION. IODOMETRlC
METHODS
Iodometry is the process of determining iodine volumetrically',
and assay method" 'which are based upon the quullt.ibtive deter-
mination of specific quantities of iodine are termed iudometric
methods. .In these methods, the renrsible reactiun 12 21-
is applied, since the reaction from left to right h---721- can be
made to go to completion in the quantitative oxidation of thiosul-
fate, arsenite, and other reducing agentR, and the reaction from
right to left h~--2I- can be made to go to cumpletion in the
quantitative reduction of permanganute, dichromate, arsenate,
ferric, cupric, and other reducible compounds. In the aSHIlY
proceSRes, either a standard solution of iodine is employed to
effect a definite chemical reaction or the iodine libemted from an
iodide is determined volumetrically by titration with a suitable
standard solution.
Iodine may function as an oxidizing agent either diredly or
indirectly. Thus, \vhen iodine dissolved in potassiuIll iodide
solution reacts with Hodium thiosulfate, it acts directly oxidizing
the thiosulfate ion to tetra.thionate ion:
12 + 2Na 2S20a----)2NaI + NU S4 02 0 .

or

The iodine combined as potassium iodide takes no part in the

oxidation, since it is already in the ionic state, but the free iodine
is reduced from the Iholecular to the ionic state by the gain of
one electron.
Iodine may act indirectly through the formation of hypoiodous
acid which is formed when iodine is dissolved in water, viz,;

12 + HOR HI + HOI
155
156 QUANTITATIVE PHARMA.C1WT!CliL CHEl,iIS1'RY

or

and

In the last reaction the arsenous acid is oxidized to arsenic

acid by the oxygen mado available from water by the reduction
of iodine to the ionic condition. Each of the two atoms of iodille
acquire one electron from one atom of !1rsenic so that the arsenic
undergoes a clmnge of charge of two when it is oxidized from the
trivalent to the pentavalent state.
The reaction 21-->1 2 can be made to go to completion by
most oxidizing agents uncleI' suitable conditions; e.g., when free
chlorine reacts with potassiulll iodide, each atom of iodine loses
one negative charge or electron and is oxidized from the ionic
to the molecular state:

or

Obviously, the iodine functions as a reducing agent, since each

atom of iodine loses one electroll which is gained by an atom of
chlorine.
Since the molecular iodine formed can be determined quantita-
tively by titration with a standard solution of a reducing agent,
the amount of chlorine which entered into the reaction can be
calculated. See assay of chlorinated lime, page 170.
Potassium iodide when dissolved in an acid solution forms
hydriodic acid and the potassium salt of the acid used; c.g.:

KI + HCl-)J(Cl + HI
When a readily reducible substance is present, it reacts with the
hydriodic acid oxidizing the ionic iodine to the molecular state:

or
2Fe+++ + 2I--2Fe++ + 12
2 X 3(+) + 2( -) = 2 X 2(+)
 OXIDATION ~lND REDUCTION 157

The ferric chloride is reduced at the expense of the iodilw.

Since each mole of Fe+++ oxidizes 1 Gm.-atolll of iodine to the
molecular state, the amount of Fe+++ which is redueed in a given
reaction can be determined by titrating the iodine oxidized to the
molecular state 'with a standard solution of a rednc'iug agent
such as sodium thiosulfate. Since the reducing agent removes
the iodine as fast as it is formed, the reaction proceeds to
completion.
Iodometric methods include some of thc most exact processes
of volumetric analysis, because, under proper conditions, the
prescnce of one part of iodine in several million parts of solution
can be recognized by means of starch indicator solution.
Starch Indicator Solution.-This solution. is prepared a:-;
follows:
Triturate 1 Gm. of arrowroot starch with 10 cc. of cold distilled
water in a mortar to form a thin paste. Heat abmit 200 cc.
of distilled water to boiling, add the starch paste to it with con-
stant stirring. Boil the mixture gently for about 30 min. or
until it forms a thin translucent liquid. Since solution of starch
deteriorates rapidly, it should be prepared freHhly each day.
Upon hydrolysis in hot aqueous solution 8tarch forms i3-amy-
lose, the soluble portion, and a-amylose, the insoluble portion.
The blue color of stareh solution with iodine is thought to be
due to the formation of a colloidal sol made up of {J-mnylose,
iodide ion, iodine, and water, Both iodide ion and iodine must
be present for the production of blue color. The blue color is
discharged in the presence of hydroxyl ion if the latter is present
in high concentration. Consequently, titl'atiolls with starch
as indicator should not be made in the presence of alkali hydrox-
ides or carbonates, but cold, dilute solutions of alkali bicarbonate
do not affect the end point appreciably.
Standard Solutions.-The standard solutiolls employed ill t.he
official assays by iodometrie methods nre 0.1 N iodine and 0.1 N
sodium thiosulfate.

Exercise 48

Object.-To Prepare and Standardize 0.1 N Sodium Thiosul-

fate,
158 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Materials Required.~26 Gm. of sodium thiosulfate.

0.2 Gm. of sodium carbonate.
2 Gm. of potassium iodide.
5 ec. of hydrochloric acid.
0.1 N potassium dichromate.
Starch indicator solution.
Procedure.-l. "Dissolve about 26 Gm. of sodiulll thiosulfate and 0.2 Gm.
of sodium carbonate in 1000 cc. of recently boiled and cooled distilled \vater."

The thiosulfate is dissolved in water previously boiled to expel

CO 2 , since the latter causes slow decomposition of Na 2S203 which
increases the reducing value of the solution:
Na2SZ0a + 2HzCO s 2NaHCO z + H 2S 20 S
H 2 S 20 S H 2SO g + S
As the sulfurous acid forms, the reducing power of the solu-
tion becomes greater, since 1 mole of the sulfurous acid formed
is equivalent to 2 moles of thiosulfate:
2Na 2S20a + 12 2Na1 + Na 2S40a
H 2SO a + 12 + H20~2H1 + H 2S0 4
The sodium carbonate acts as a stabilizing agent by converting
the thiosulfuric acid formed into sodium thiosulfate.
2. "Measure accurately 30 cc. of tenth-normal potassium dichromate
into a glass-stoppered flask and dilute it with 50 cc. of distilled water. Add
2 Om. of potassium iodide and 5 cc. of hydrochloric acid, stopper and allow
to stand for ten minutes. Dilute with 100 cc. of distilled waier and titrate
the liberated iodine with the sodium thiosulfate solution. When the solu-
tion has assumed a yellowish-green color, add starch T.S. and continue with
the titration to the discharge of the blue color. Calculat.c the normality of
the sodium thiosulfate solution and, if desired, adjust exactly to tenth-
norllllll."

The acid solution of potassium iodide quantitatively reduces

chromic acid to green chromic chloride, liberating an equivalent
quantity or iodine:
K 2C1'207 + 6K1 + 14HCl~2CrCla + 8KCI + 7H 0 + 31 2 2
294.2 6(166.02) 6(126.92)
It is evident from the above equation that 1 mole, 294.2 Gm.,
of K2Cr201 reacts with 6 moles, 996.12 Gm., of K1 to oxidize
761.52 Gm.-atoms of iodine from the ionic to the molecular state.
 OXIDATION AND REDUCTION 159
Potassium iodide must be present in excess to pre"vent thp
precipitation of iodine which is insoluble in aqueous solution but
soluble in KI solution.
The solution is diluted to a large volume, 350 ce., to make the
end point more easily distinguishable, for the latter il; not indi-
cated by the disappearance of color but by a change in color
from blue to light green.
Since tenth-normal oxidizing and reducing solutions are equiva.,
lent cubic centimeter for cubic centimeter, the calculatiOll of
the normality of the thiosulfate solutioll is as follows:
cc. 0.1 N K 2Cr 2 0 1 X 0.1 = normal't1 y 0 f th e so d'Hllll thO108UIfa t e
cc. N a2 S20 3
solution.
Sodium thiosulfate solution changes in its reducing power
due to the action of carbon dioxide, sunlight, and dust. The
solution should, therefore, be preserved in tightly stoppered
bottles and be kept away from direct sunlight.
Sodium thiosulfate solution may also be standardized against
pure iodine dissolved in potassium iodide solution or against
iodine set free from acidulated solution of potassium iodide by
standard solutions of potassium permanganate, potassium bro-
mate, or potassium iodate.

Questions and Problems

1. Illu~tl'lLte how iodine l11!1y function directly and indirectly as an oxidiz-
ing agent.
2. How may CO 2 change the reducing power of standard sodium thio-
sulfate solution? Use reactions to illustrate the explanation.
3. When potassium dichromate is reduced by iodinc, what change ill
polarity doe~ each Cr atom undcrgo? Is the iodine oxidized or reduced?
4. Why must 1(1 be present in excess before the R,Cr,Or is added?
5. Wh[lt purpose does the Hel serve?

Exercise 49
Object.-To Prepare 0.1 N Iodine Solution.
Materials Required.-13 GI11. of pure iodine.
36 Gm. of potassium iodide.
Procedure.-"Weigh accurately about 12.75 Gm. of reagent iodine and
transfer it quickly into a solution of 36 GI11. of potassium iodide in 100 cc.
of distilled water. After solution is complete, dilute to exactly 1000 ce. at
160 QUrlN'l'I'l'A'I'JVE PIlAllM.ACEU'I'IC'AL CIlEMIS'l'RY

25C. From the weight of the iodine used calculate the normality. The
normality of the solution should frequently be redetermined."

One gram of iodine is soluble in 2,950 cc. of water. In the

presence of sufficient potassiuHl iodide, however, its solubility
is greatly increased due to the formation of an unstable, soluble
polyiodide of potassiulll, lOa:

KI + 12 KI3
In the presence of reducing agents, the reaction proceeds quanti-
tatively to the left, and the solution reacts like a solution of
iodine alone.
The iodine should be weighed accurately in a glass-stoppered
weighing bottle, transferred quantitatively to the solution of
36 Gm. of KI in 100 ceo of distilled water, and after it is com-
pletely dissolved, the solution should be made up to exactly
1,000 cc. The solution of potassium iodide used to dissolve
the iodine is of sufficient concentratiOll to dissolve the iodine
rapidly so that no appreciable loss will occur due to volatilization.
If pure rcagent iodine is used in preparing this solution, it will be
approximately tenth-normal and need not be standardized. Due,
however, to the difficulty of weighing exact quantities of iodine,
it is usually more convenient to prepare a solution of approximate
strength and standardize it against 0.1 N sodium thiosulfate
solution as directed in the following exercise.
Exercise 1i0
Object.-To Prepare and Standardize 0.1 N Iodine against
Sodium Thiosulfate.
Materials Required.-14 Gm. of pure iodine.
36 Gm. of potassium iodide.
0.1 N sodium thiosulfate.
Starch test solution.
Procedure.--Dissolvc about 14 Gm, of rengent iodine in a solution of
36 Gm. of potassium iodide in ahout 100 ce. of distilled water, diluting
finally to 1,000 ceo Carefully measure from a burette 25 cC. of this solution
into a flask, then add gradually and cautiously from a burette 0.1 N sodium
t.hiosulfate, shaking constantly, until the color of t.he solution is but slightly
yellow, t.hen add a few drops of starc~h T.S. and continue the addition of the
0.1 N sodium thiosulfate until the blue color is just discharged. Note the
number of cubic centimeters of t.he 0.1 N sodium thiosulfate consumed, and
 OXIDATION "iND REDUCTION 161
then dilute the iodine solution so that 25 ce. will require for deeolorizl1tioll
exactly 25 ce. of the 0.1 N sodium thiosulfate at standard tc'llJperatllfe.

Although the U.S.P. directs that reageat iodine he uflml in

preparing 0.1 N iodine solution, iodine "containing ehlorillP in
small amounts as impurity could he used, since the ehlorine
reacts with potassiulll iodide to liberate iodine:

Iel + KI KCl + 12
The sodium thiosulfate reacts with the iodine to form sodium
iodide and sodium tetrathionate, and it Vf'ry slight excess of
thiosulfate discharges the blne color of the indicator:

I~ + 2Na2S20a 2NaI + NU28406

If 25 cc. of approximately 0.1 N iodine require exaetly 25 cc.
of 0.1 N sodium thiosulfate, the iodille 11'1 exactly tenth-normal,
but, if only 24 ce. of 0.1 N iodine solution are n~quired, the
solution is fitronger than normal and each 2.t (:('. of iodine solu-
tiOlllllltst he diluted to 25 cc. to secure an exactly 0.1 N solution.
Since 0.1 N iodinc solution deteriorate::;, it will not remain
precisely tenth-normal very long. It is therefore best to calculate
the normality of the solution and empluy it ail a standard l:iolution
of known value. In the aboye ease, the approximately 0.1 N
iodine solution would be 25/2,1 = 1.0417 times tenth-llormal
and should be labeled 0.1042 N iodine. vVhenever the solution
is used in it determination, the numher of cubie eentimeters of
0.1 N iodine employed mit)' be ealeul!tted; i.e., cubic eerttimeters
approximately 0.1 N iodine X 1.0417 = eubie centimeter::;
0.1 N iodine.
Iodine solutions may a,lfio be standardized Rgainst pure arsenous
oxide (fiee Exercise 51).
Questions and Problems
1. Why mllY iodine conta.ining chlorine or bromine as impurity be used to
prepare standard solutions of iodine when the solution is standardized
against sodium thiosulfate?
2. What is the normality of a solution containing 1.5420 Gm. of iodin!!
in 100 ce.? .
3. If 24.25 ce. of iodine solution requires 25 ce. of 0.0858 N Na2S20~1 what
is the normality of the iodine solution?
162 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

OXIDATION-REDUCTION METHODS BY DIRECT TITRATION WITH

STANDARD IODINE SOLUTION

Exercise 51
Object.-Assay of Arsenic Trioxide.
Materials Required.-About 0.2 Gm. of arsenic trioxide.
Sodium hydroxide test solution prepared by dissolving 4.3 Gm. of NaOH
in sufficient distilled water to make 100 cc.
Diluted sulfuric acid.
2 Gm. of sodium bicarbonate.
0.1 N iodine.
Starch indicator solution.
Procedure.-l. Dissolve about 0.2 Gm. of arsenic trioxide previollsly
dried to constant weight at lOOoe., and accurately weighed, in 20 cc. of
boiLing distilled water, and graduaIIy add sodium hydroxide T.S. until
complete solution results.

Arsenic trioxide is slowly soluble in cold water, more rapidly

soluble in boiling water, and readily soluble in sodium hydroxide
solution with the formation of sodium arsenite:
AS20~ + 6NaOH~ >2Na aAs0 3 + 3H 02
197.82

Sodium arsenite on hydrolysis yields sodium hydroxide and

arsenous compounds to form a distinctly alkaline solution.
2. Neutralize this solution with diluted sulfuric acid, using phenol-
phthalein T.S. as indicator, cool, dissolve in it 2 Gm. of sodium bicarbonate,
and titmte the mixture with 0.1 N iodine, using starch T.S. as indicator.
Each cnbic ceDtimeter of 0.1 N iodine corresponds to 0.004946 Gm. of
As 20 3

If iodine is added to this alkaline solution, some of it tends to

form sodium hypoiodite, NaIO, or similar compounds which do
not react readily with arsenous acid:

2NaOH + k---l-NaIO + NaI + H 2 0

The excess sodium hydroxide, therefore, required to dissolve the
As 20 S is exactly neutralized by sulfuric acid. Sodium bicarbon-
ate is added to neutralize the HI formed in the reversible reaction
HaAsOa + H 0 + 12
2 HaAsO~ + 2Hl
 OXIDATION AND REDUCTION 163

The NaHCO a removes the HI aR rapidly as it is formed, causing

the reaction to go to completion toward the right:

Under the conditions of the assay, :mdiUIn bicarbonate does

not react with iodine, but, if an excess of iodine is added, it may
react slightly. The reaction between arsenous ltcid and iodine
proceeds quu,ntitatively to completion best in a neutral solution.
Since HI is formed in the reaction, a buffer must be added to keep
the solution neutral. The sodium bicarhonate in this case is
the buffer.
The oxidation of a mole of arscnic from As+++ to AH++-H-I-
requires 2 Gm.-atoms of iodine. The change of polarity of
As+++ when oxidized to As+++++ is 2( +), and since each mDle of
As 20 a contains 2 atoms of As, the total ehallge of polarity is
2 X 2(+) = 4( +). One cubic c(mtimeter of 0.1 N iodine is
therefore equivalent to 4X
i~7:;,ooo = 0.004946 Om. As 2 a, or 0
74.96
2 X 10 X 1,000 = 0.003738 Gm. As.
U.S.P. arsenic trioxide should contain 99.8 per cent of As 20 a
Calculate the per cent purity of the As 20 a aSHayed and the per
cent of arsenic which it contains. Many arsenous compounds,
antimonous compounds, etc., and their preparations may be
assayed by similar procedure.

Questions and Problems

1. Writc.the reaction between AS~03 and NaOH structurally.

2. Why must the exceRS sodiuJIl hydroxide be neutmlized with !Leid before
tho titration with st!LIul::ml iodine solution?
3. What substance is employed us !L buffer in the assay? Why is It buffer
necessary?
4. Name some other official substances or preparations containing anti-
mOllous or arsenous compounds to which the above methOli is applied.
5. If pure As 20 S is avaihlble, how might a solution of iodine be standard-
ized against it?
6. Ascertain the official method of assay for anthnonous oxide, explain
each step in the procedure, write equations for the reactions involved, and
calculate the equivalent weight of 8b 20 3
164 QUANTI'l'il'l'IVE Pl!ARMACEU'l'[Ci1L CHRMISTRY

TABLE XVIII.-OFFWIAL SUBSTANCES AI:;HAYED BY DIRECT TITRA'i'ION WI'l'H

0.1 N IODINE

Alnount Equiva-
used, lent of Official requirClnellt p
Substance
Gm. or 1 CC. per cent
CC.

U.S.P.
.Antimony and l)OtuB.8iuJD tartrate . ... . 0.5 0.01670 I((SbO)C.,n,Oo.).fH.O ~ 99
Al'senie h:Uodidc .... .. , ........... ~ . 0.5 0.02278 AsIa '" 99
Arsenic trioxid(~ .. , .... , , . , ......... . 0.2 0.004046 As,O, = 99.8
Arsenous [lUill, solution . ........... , . 20 0.OO401G o,
A = O.H7G to 1.025 TV IV
Potassiutn {lfHenite, solution ..... . , . " 20 0.004946 A.,O, = 0.975 to 1.025 W IV
Sodium tlliORl1lCate ................. . 0.5 0.01581 Na,S.O, ~ 99
Trypnrsumiuo ... ....... , .......... . o ., ll. 003746 As = 25.1 to 25.5
N.F.
Arsenic linu mercw'ic iorlio(!, solution
(for AsIa) ....................... . 25 0.02278 As!" = 0.95 to 1.05 WIV
Rodium rJlwodylutc, Ilmpul~, .. , ...... . 0.2" 0.007008 Na(CII,),AsO, = 69 to 76 b
Sodium tlliosu1iate, IlWpUL9, ...... , .. . 1" 0.01581 Ni1.S20, '" 60.5 to 66.9"

a Amount, of ingredient sought.

b Pel' cent of labeled amount.

OXIDATION -REDUCTION METHODS BY DIRECT TITRATION OF

FREE IODINE WITH STANDARD SODIUM THIOSULFATE
SOLUTION
Exercise 52
Object.-Assay of Compound Solution of Iodine for Iodine
Content.
Materials Required.-5 ce. of compound solution of iodine.
OJ. N sodium thiosulfate.
Starch inaic[Ltor solution.
Procedure.-"Dilutc exantly 5 cc. of Compound Solution of Jodine with
25 ce, of distilled W[Ltcr, and titrate with tellth-normlll sodium thiosulfate,
stan:h 1'.S. being used us the.indic!ltor. Each cubic centimeter of tenth-
llorniil.l sodium thiosulfate is equivalent t.o 0.01269 Gm. of I."
The eompound solution of iodine is diluted with water to bring
it to approximately tenth-normal. This dilution (Jan be made
without the addition of KI to prevent precipitation of iodine,
since the compound solution of iodine contains 10 per cent of KI.
The starch solution should not be added until the titration mix-
ture has acquired a straw color. The following reaction takes
place':
12 + 2Na S 03.5H 0-Na S400 + 2NaI + lOH 0
2 2 2 2 2
2(126.\)2)
 OXIDATION liND REDUG'TlON Hifi
Each cubic centimet.er of 0.1 N sodium thiosulfate consumed
. tI t't t" . I t 253.84 1 G
III Ie l.ra lOll lS eqUl\'a en to 2 X 10 X 1,000 = 0.0 2692 .:rm.
ioeline. The U.S.P. requires that eompound :-;olution of iodine
contain not less than 4.5 or more than 5.5 Om. of iodine ]Wl'
100 cc. of solution. Note that this i:-\ a \\'C'jght-to-yolume per-
centage. Calculate the per cent of iodine in the sample u~sllY('cl
and compare the result with the official requirement.
Elemental iodine, tinctures of iodine, and any preparation
containing free iodine may be as~myml hy methods similar to
that given above, provided that other snbl';tauccs which oxidize
sodium thiosulfate are absent.

Questions and Problems

1. Why can compound solution of iodine he diluted witho1lt :ulding I\I
to prevent the precipitation of iodine?
2. Give it procedure for the assay of nlemental iodine with re!I.~(Jns for
each step. Write eq1Ulijolls for aU of the reactions iIlVO\vell.
3. Name seveml 8ubstaneeH which, if present, would o::-;idi?,(' )wdiulll
thiosulfate.
4. Why is it best not to add the starch indict,tor solution lwfo1'I' the iodine
solution has acquired It tl1.rnw color?

TABLE XIX.-OFFICIAr~ SURS1'ANGES ASSAYED BY DLRE(,vl' TI'l'RATlON Wl'l'f{

0.1 N N::t,S,O"

AlUollllt,
Equiva-
used, Oflidal reqllirement,
Subst!lnee lent. of
Om. per ccut
1 ec.
or ce.
---- - - "
U.S.P.
Iodine ...................... 0.5 0.0126() 12 = 09.5
Iodine, compound solution of. 5.0 0.012li9 I2 = 4.5 to 5.5W/V
Iodine poutoxide, R .......... 0,5 0.002782 1.0. = U9.5
Iodine, tinetul'c of .. , ........ 5.0 0.01269 12 = u.5 to 7.5W/V
Iodino, tincture of, mild . ..... 5.0 0.(1261) I~ = 1.8 to 2.2H'/V
N.F.
Iodine, ampuls of ............ o.175a 0.01269 12 = 95 to l05W /Vb
Iodine, stronger tincture of ... 2.0 0.01269 I. = 16 to 17W,IT"
Iodie acid, R ............... , 0.1 0.002932 HIO s = 99.S

R. = reagent .
. Amount of ingredient sought.
Per cent of the labeled amount.
166 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

OXIDATION-REDUCTION METHODS INVOLVING THE USE OF

EXCESS STANDARD IODINE SOLUTION AND RESIDUAL TITRA-
TION OF THE EXCESS IODINE WITH STANDARD SODIUM
THIOSULFATE SOLUTION
Exercise 63
Object.-Assay of Mercurous Chloride.
Materials Required.-About 0.7 Om. of mercurous chloride.
50 cc. of 0.1 N iodine.
5 Om. of potm;siUlll iodide.
0.1 N sodium thiosnlf;L1,e.
Stlll'f~h imlicrltOl' solution.
Procedure.-l. "Dry about 0.7 0111. of Mild Mercurous Chloride t.o
constant woight over sulfuric acid, weigh accurately. mix well with 10 cc.
of distilled water in a. glass-stoppered flask, lLud add 50 ee. of tenth-normal
iodine and 5 GIll. of potassium iodide dissolved in 10 ce. of distilled water."

Since mercurous chloride is practically insoluble in water, the

purity of the salt cannot be determined by the precipitation
method used to determine the chloride ion in a soluble chloride.
"When mercurous chlorido is treated with a solution of iodine in
potassium iodide, it dissolves quite readily with the formation
of a double salt of mercury and potassium iodides:
2HgCl + 6KI + h---72HgK I + 2KCl 2 4
236.07
2. "St.opper the flask, allow the mixture to I:lialld with occasional agitation
until complete solution has taken place, and then titrate the residual iodine
with tenth-normal sodiuIll thiosulfate, stareh T.S. being llsed as indicator.
Each cubic centimeter of tenth-llormal iodine is equiv[1lent to 0.02361 Gm.
of HgCl."

In the assay process, the mercurous chloride mixture is allowed

to stand with occasional agitation until the reaction has gone to
completion as indicated by its complete solution. The excess
0.1 N iodine added is then determined by residual titration. The
formula of the double salt of mercuric potassium iodide may be
written, HgI2.2KI, and the ionic reaction may be represented as:
2Hg+ + I2~2Hg++ + 21-
The number of cubic centimeters of 0.1 N iodine found upon
residual titration with 0.1 N sodium thiosulfate subtracted from
the number of cubic centimeters originally added gives the
 OXIDATION AND REDUCTION 167
volume of standard iodine solution constlmed ill the oxidation of
the mercurous ion to mercuric ion. Since 1 Gm.-ioll of iodin!' j"
equivalent to 1 Gm.-mole of HgCI, each cubie eentimetcr of
o.1 N'lO(rme consmnelI 1S. eqUlva
. 1ent t0 ITi236.07
X 1,000 = 0.023 6 1
Gm. HgCl.

Questions and Problems

1. Why is the mixture of HgCl with solution of ioditlc in IU "topperetl
lIud allowed to stand?
2. Indicllte by ionic reactioIls hoI\' iodine fUllctions flS till oxidizing agent
in the i1HS!\y of sodiulll sulfite (U.S ..P. reagent) and sulfurous acid (U.S.P.
reagent).
3. ExpltLin !l:wh step involved in the oflieial assay of menmrous iodide
[IUd write ionic ally all of the refwtiollS which opcur during the assay.
4. If an ullknown sample treated with 50 ec. of 0.D50 N indine required
18.6 of 0.1025 N sodium thioslili'ate to titrate the execs:'; iodine, to what
amount of each of t.he following substances would the sample be equivalent:
CR,COCR" JJgCl, Na2S.DH,O, find R0 2 ?
TABLE XX.-OF~'ICIAL SUB>l1'ANCES ASSAYED BY RESIDUAl, TITRATION OF
ExCESS STANDAIW IODIKE SOLUTION WITH 0.1 N Nn~S20"

Amount
El[uinl-
used, Oflicial fPqlliremcnt,
SubstllllC(l lent of
Gill. or p{;r cent
1 ce.
ce.
-----------------------------1--------1------------------
U.S.P.
Acetone ................. . l.0 0.0009675 CH,COCH, = 99
Mercurous chloride ....... . 0.7 0.02361 HgCI = 99.6
Merc:urolls iodide ......... . l.0 0.03275 HgI = 99
Methylthioninc chlori(10 .. . 0.12 0.005328 C,,,H 1 RN.CIS = 98.5
Sodium bisllifite, R ....... . 0.005203 NIlHSO, = 90
Sodium oulfite, R ......... . 0.25 0.000303 NaISO. = 95
Sulfurous add, R ......... . 2.0 0,003203 S02 = II
N.F,
MercurouH chloride, mild,
and sodium hicn.rbonatc,
tablets of. .... , ....... '. 0 .3a O. 02361 EgCl ~ 92.5 to 107.5"
Mercurous chloride, mild,
tahletsof ...... , ........ 0.3" 0.02361 RgCl "" !JZ.5to I07.5h

R. = reagent .
Amount of ingrodient sought.
b Per cent of the labeled amount.
168 (JTJA_NTITA.TIVE PHAWHACl!JUTlCA.L CHEMISTRY

OXIDATION-REDUCTION METHODS INVOLVING THE LIBERATION

OF IODINE FROM POTASSIUM IODIDE AND TITRATION OF
THE FREE IODINE WITH STANDARD SODIUM
THIOSULFATE SOLUTION

Exercise 54
Object.-Assay of Solution of Ferric Chloride.
Materials Required.-About 2 Gm. of solutioIl of ferric chloride.
5 ce. of hYllrochlorie 11cid.
3 Gm .. of potassium iodide.
0.1 N sodium thiosulfate.
Starch indicltt.or Roiution.
Procedure.-L Transfer nbout 2 Gm. of a solution of ferric chloridll to
a tared flask, stopper, and weigh aecurately i add 5 ce. of hydrochloric acid,
25 ce. of ciist.ilh,n water, and about 3 Gm. of potassium. iodide. Securely
stopper the flask, Ilnd allow the mixt.ure to stand for 5 min.

The solution of ferric chloride may more conveniently be

weighed in a glass-stoppered weighing bottle. Loosen the stopper
of the weighing bottle and place the bottle, stopper, and contents
in a 250 ce. Erlenm~;yer flask: containing 5 ceo of HCI and 25 cc.
of distilled water. Remove the stopper from the weighing bottle
with a stirring rod and thoroughly mix the solutions, add the
IeI, and stopper the flaHk securely. The solution is allowed to
stand for 5 min. to permit the following reactions to go to
completion:
IG + HCI~ KCl HI +
2FeCh + 2HI ---).2FeCb + 2HCl + 12
162.21
or

Sufficient KI must be present to form enough HI to reduce tlH~

ferric ion quantitatively to ferrous ion and to dissolve the free
iodine formed.
2. Dilute the liquid, with 50 ce. of distilled water, and then titrate the
liberated iodine with 0.1 N sodium thiosulfate, starch T.S. being used as
indicator. Each cubic centimeter of 0.1 N sodium thiosulfate corresponds
to 0.005584 Gm. of Fe.
 OXIDA.TWN AND REDUC''l'ION 169
The liberated iodine is titrated with 0.1 ]I[ sodium thiosulfate
in the usual manner.

Each mole of Fc-r:.-~ is equivalent to 1 Gm.-atolll of iodine.

Each cubic centimeter of 0.1 N Na 2S203 is therefore equivalent
to 101~21~~OO = 0.016221 Gm. FeCla or 10 ~~~ooo = 0.005584
Gm. Fe.
According to the U.S.P. definitioll, solution of ferric chloride
should contain not less than 10 per cent or more than 11 per cent
of Fe. Calculate the per ccnt of Fe and the per CCllt of FcCh
in the sample assayed and compare the results obtained with the
official requirement.
TABLE XXI.-OFFICIAL SUBS'rANCES ASSAYED FOR Fe BY Trl'RA'l'IOX OF nIE
IODINE LmBRATED 1'lWM IU WITH 0.1 N Na,S,Oa

Ali.~ount IEquiva-
1,,~d. I<mt of Ofti(dnl requil{_~nH'nt,
Substanee
Gm. or I 1 I~C. pCI' eent
(~C.

U.S.P.
Ft~rrie chlori<Ie, solution of ....... ..... _
- --- -
2.0
iF".\' _- 10 to II
0--. (),),.)-,C)84
_
Ferric chloride, tincture o, ......... ,.. 5.1l 0.005584 Fe = .1.5
Ferric citmte, R............ ..... ..... 1 0.005584 Fe = Hl.5 to 18.5
Ferric "ulf:1lc, solution of.............. 1.5 0.005584 Fe = 9.5 to 10.5
Iron und ammonium citrntes ...... ..... ! 1.0 O. 00558~..I: Fe = 10.5 to 18.5
Iron and Itnullonium citrates, green ..... [ I 0.005584 Fe = 14.5 to 16
N,F.
Ferric citrochloride, tincture of......... 5.0 0.005584 Fe .. 4.481V/V
Ferric glyceropho"phu to.. . .. .. .. .. .. .. 0 .5 0 . O()5584 Fe = 11
Ferric hYl'ophosphitc. . . . . . . . . . . . . . . . . . 1.0 O. 005li84 Fe = 21.8
Ferric oxide. Slwcharated............... 8.0 0.005ii84 Fe = 2.8 t,o 3.2
Ferric phosphate. soluble.............. 1.0 0.005584 Fe = 12 to Iii
Ferric pyrophosphate. soluble.......... 1.0 D.OO5fJ84! Fe = 10 .5 to 12.5
FerricsubsuUa[e,Bolution of..... ...... 1.0 0.005584 l!'e = 20to 221F/Y
Iron and ammollium ncetate, solution of. 26 0.005584 Fe = 0.16 to O.201V/V
Iron, and ammoniuIll cit,rates, green,
ampul. 01. . . . . . . . . . . . . . . . . . . . . . . . . 1a 0.005084 Fe = 14.5 to W.O'
Iron, peptonized.. .. .. . .. . . . . . . . . . . .. . 0.5 0.005584 Fe ~ 10 to 18
Iron" peptonized, and manganese. solu-
tion Of ........ , .................. 1 25 0.005584 Fe = 0.265 to 0.325 W/Y
Iron, peptoni;!,ed, solution of. . . . . . . . . . . . 25 0.0(155841 Fe = 0.21)5 tQ 0.325 W/V
1

R. ;" reagent .
Amount of ingredient sought
Per cent of the I.beled amQunt.
170 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The principle illvolved in th(~ method of assay in this exercise is

applicable to most official determinations of ferric salts with slight
modifications, e.g.,. solution of iron and ammonium acetate,
solution of ferric sulfate, soluble ferric phosphate, solution of
ferric citrate, albuminized iron, peptonized iron, ferrous lactate,
and others.
Questions and Problems
1. Explain how pottl~sium iodide in flcid solution functions as a reducing
agent.
2. Ascertain the method of assay of peptonized iron in the N.F. and
apply the principles studied in the above assay in an expittnation of each
step of the procedure.
S. Caleulate the amount of ferric sulfate, ferric oxide, nne! ferrous lactate
equivalent to 1 ce. of 0.1 N KMn01.

Exercise 55
Object.-Assay of Chlorinated Lime.
Materials Required.-About 4 Gm. of chlorinated lime.
1 Gm. of potassium iodide.
5 cc. of ncetic acid.
0.1 N sodium thiosulfate.
Starch indicator solution.
Procedure.-l. "Transfer to a mortar ahout 4 Gm. of Chlorinated Lime,
accurately weighed in a tared weighing hottle, using 50 ee. of distilled water.
Triturate thoroughly, and pour the mixture into a 1,000 ce. graduated flask,
rinsing the mortar with distilled water to make 1,000 cc. Stopper the flask
and allow it to stand for ten minutes."

Tho accurately weighed sample of chlorinated lime is triturated

with water to disintegrate it to a fine powder.
2. "Shake thoroughly, add to 100 ce. of the mixture 1 Gm. of potassium
iodide and 5 ce. of acetic acid, and t,itrate the liberatcd iodine with tenth-
normal sodium thiosulfate, starch T.S. being used as the indicator. Elteh
cubie centimeter of tenth-normal sodium thiosulfate is equivalent to 0.003546
Gm. of available chlorine (C1)."

An aliquot portion of the mixture representing 0.4 Gm. of

sample is treated with 1 Gm. of KI and 5 ce. of acetic acid. In
the presence of acids the available chlorine is liberated from the
chlorinated lime. Available chlol'ine is usually defined as the
chlorine that is liberated from a substance by acids. 1'he free
 OXIDATION AND REDUCTION 171
chlorine reacts with K1 to Hber[Lte iodine C[lmntitrLt.ively, and
the quantity of iodine set free is determined by titration \vith
0.1 N Na2S20S.
Chlorinated lime is thought to be acalciul1l chlol'o-hypoehlorite,
Ca(OCI)CI, which may he regarded as a compound contai'ning
equimolecular portions of calcium hypochlorite, Ca(OCI)~, and
calcium chloride, CaCI 2 :

2Cu(OCI)CI---+CaC1 2 + Ca(OCl)2
All of the chlorine present in cakiulll nhloro-hypoehloritc IS
released when the compound is treated with acetic [wid:

C[L(OCl)CI + 2CH sCOOH---+Ca(CH COO)2 + H 3 0

2 + Cb
The liherated C1 2 reacts wi. th K1:

C1 2+ 2K1---+21\:C1 12 +
2(35.46) 2(1213.92)

From the [Lbovc reaetionR, it is apparent that 2Na2S20;! i!-l equiva-

lent to 12 which is equivalent to C1 2
One cubic centimeter of 0.1 N N a 28 20 3 is equivalent to
35A) () ()OgI::46
f6-126.92 0 0 692
X 1,000 = . 12
G . l'
me
lll1. lOC
1
nIlC 10 X T,511O =. a

Om. chlorine.
Other substances which yield free chlorine or free bromine
when trea.ted with acids may he u,;;sayed in the same way, e.g.,
solution of sodium hypochlorite.

Questions and Problems

1. What is "Iwailable chlorine"?
2. What funetion does KI RCfve in this assay? Must the K1 be present
in excess?
3. If a 0.4 Gm. sample is known to C',ontllin 32 per cent of avniJahll1
chlorine, how much J\:1 would be necessary to yield an llmount of iOlline
equivaJent to the chlorine?
4. Look up the U.S.P. requirement for chlorinated lime and compare it
with the assay result.
172 QUANTITATIFE PHARMACEUTICAL CHEMISTRY

TABLE XXII.-OFFlC'IAL SUIlSTAN0ES ASSAYED FOR AVAILABLE CHLORINE

BY TITRATION OF 'fHE IOIJINE LlBERA'l'lm l'HOM KI WITH 0.1 N Nu 2S20,

Amount
Equiva-
used, Official requirement,
Substance lent of
Gm. or per cent
1 cc.
CC.

U.S.P.
ClI1orarnine-T .... , , . , . , 0,2 0,001773 Cl (active) = 11.5 to 13
Dich1oraminc-T., . , , .. , 0,1 0,001773 Cl (active) = 28 to 30
Chlorinat.ed lime, R .. , . , 4 0,003546 Cl (available) = 30
Sodium hypochlorite,
solution of. .......... 5 0,003723 NaOCI = 4 to 6
Sodium hypoc h10ri to,
solution of, diluted, . , 25 0.003723 NaOCI = 0.45 to 0.50W /V
l{. = reagent.
Exercise 56
Object.-Assay of Cupric Sulfate.
Materials Required.-l Gm. of cupric sulfate.
4 ce. of acetic acid.
3 Gill. of potassium iodide.
0.1 N sodinm thioRulfate.
Starch indieator solution.
Procedure.-" Dissolvtl about 1 Gm. of Cupric Sulfate, accurately
weighed, in 50 cc. of distilled water, add 4 cc. of acetic acid and 3 Gm. of
potassium iodide, and titrate i,he liberated iodine with tenth-normal sodium
thiosulfate, starch T.S. being used as indicator. Each cubic centimeter
of tenth-normal sodium thiosulfate is equivalent to 0.01596 Gm. of CuSO~."
'This aHsay is based on the reaction between cupric sulfate
and potassium iodide in which the copper is precipitated as
cream-colored cuprous iodide and one atom of iodine is liberated
for each cupric ion present:
2CUS04 + 4KI-Cu212 + 2K S0 + 12
2 4
2(Hi9.63)
or
2Cu H + 41----t2Cn+ + 21- + 12
Each cubic centi.meter of sodium thiosulfate consumed is
126.92 159.63
equivalent to 10 X 1,000 = 0.01269 Gm. 12 , to 10 X 1,000
63.57
"" 0,01596 Gm. CUS041 and to 10 X 1,000 = 0.00636 Gm. Cu,
 OXIDATlON AND IVJ:DU(:'1'lON 178
U.S.P. copper sulfat.e should contain not less than 63 per Cf'llt
01'more than 66.8 per cent of CUSO.l corresponding to 98.5
per cent of CuS04.5H20. Calculate the per cent eu, CUS04
and CuS04.5H 20 in the Barnple assaypd.

Questions and Problems

1. What funct.ion does the acetic !toid serve ill t,his (Lssay?
2. Explain the reduct.ion of copper from tho cupl'ie to t.ho euprou:l con-
dition in accordance with the ionic theory. Indicate all tnlllSfcl's of ('hargu.
3. How much iodine will 0.25 Gm. of CuSO. liberate from KI in aoici
solution?

Exercise 6'1'

Object.-Assay of Exsiccated Sodium Al:senate.

Materials Required.-O.3 Gm. of exsit~cated ~ollilllll UiscIllLte.

10 ce. of hydrochloric !leid.
3 Gm. of pot.assium iodidt'.
0.1 N sodium thiosulfate.
Starch indicator solution.
Procedure.-" DissolYl about 0.3 Gm. of Exsiecated SodiUlIl Arsenate,
dried to con~tllnt weight at 150"C. and accll1'ately weiglH'd in 25 cc. of dis-
tilled water in a glaHs-stoppereLl fia~k;.,hcat the solution to 80"C., and add
10 cc. of hydrochloric aeid alld 3 Gill. of potassiulll iodide. Allow the mix-
ture to !ltand in the stoppered flask for 15 minutes ut 80C.; then eool, and
titrate the liberated iodine with tenth-norm!ll sodium thiosulfate, using
starch T.S. a1l the indieator. Conduct a hlank te>lt, and dedlld the amount
of tenth-normal sodium thiosulfate consullwd in the blank. Each cubic
centimeter of tenth-normal sodium thiosulfate is equivalent to 0.009295 Gm.
of Na'2HAsO . " "

Sodium arsenate in strongly acid solution liberates iodine from

KI with reduction of As+++++ to As+++, the reaction going to
completion when the solution is warmed and allowed to stand.
The 'reactions which take place, although probably more complex,
may be represented as follows:

Na 2 HAs0 4 + 2HC]---)H aAs0 4 + 2NaCl

185.91
+
HCl + KI---4-1(Cl HI
HaAs04 + 2I-U-HaAsOa + H 0 + h 2

2Na2S~03 + I~---4-2NaI + Na2S 06 4

174 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Since each mole of Na2HAs04 liberates 2 Gm.-atoms of iodine,

1 cc. of 0.1 N Na 2S20 a is equivalent to 2 X i~5~~,000
0.009295 Om. Na2HAs04.
N.F. exsiccated sodium arsenate contains not less than 98
per cent of Na 2HAs0 4 Calculate the per cent purity of the
sample assayed and compare the result with the official require-
ment. Also calculate the pCI' cent of arsenic in the sample
assayed.
Arsonic and antimonic compounds may be assayed by methods
similar to that given above. The arsenic and antimony present
in organic compounds in the trivalent state may also be aHsayed
by this method after quantitative oxidation to the pentavalent
state. Such procedure is employed in the official assays of
arsphenamine and neoarsphcnamine.

Questions and Problems

1. Assuming that the sample uAed in the assay was pure Na 2HAsO,.7H 20,
how much KI would be required?
2. Write the equations involving oxid(1tion"reduction ionically.
3. N!1me several official an;cllic !md !tutimonic compounds assayed by
methods which employ the same basic principles.
4. What change in valence does the arsenic undergo in the nbove assay?
Does the change of valence correspond with the change of charge'/

TABLE XXIII.-OFFICIAL SuaS'l'ANCES CONTAINING ARSENIC ASSAYED BY

TITRATION OF THE IODINE LIBERATED FROM KI WITI! 0,1 N Na 2S 20a

Amount
Equiva"
used, Official requirement,
Substance lent of
Gm. or pel' cent
1 cc.
co.
---- - - " -
U.S.P.
Arsphenamine ..... 0.2 0.003746 As = 30
N eoalsphenamine .. 0.2 0.003746 As = 19 to 22
N'.F.
Sodium nrsenate,
exsiccated .... " .. 0.3 0.009295 Na 2HAs0 4 = 98
Sodium nrsenate,
solution of ....... 25 0.009295 Na2HAsOj = 0.975 to 1.025W IV
 OXIDA.TION A.ND REDUCTION , 175

Exercise 68
Object.-Assay of Thyroid.
Materials Required.-l Gm. of thyroid.
A nickel crucible.
9 Gm. of anhydrous potassium carbonate.
7 Gm. of anhydrous sodium cttrhonato.
5 Gm. of potassium nitmtc.
50 CC. of solution of chlorinated soda.
About 60 cc. of diluted phosphoric acid (1 to 1).
0.1 Gm. of pot.assium iodide.
0.005 N sodium thiosulfate.
Starch indicator solution.
Procedure.-l, "Thoroughly mix 1 Gm. of Thyroid, finely pow(l(ln~d am:I
accurately weighed, with 15 Gill. of an intiIllltte mixture of 138 pn.rt;; by
weight of anhydrous potassium carboIHlte, 106 parts of anhydrous sodium
carbonate and 75 parts of powdered pot!lsRium nitrate in a nickel crucible
of about 125-cc. capacity, and spread an additional 5 Gm. of this JIlixture
evenly over tho surface. Heat the crucible with the flame of a Bunsen
burner at such a rate as to attaiIl a dull red eolor in ten milHltes, and conl:illUC
the heating at the same tempernture for an additional ten-minllte period.
At the end of this time the carbonaceous materhtl is completely oxidized
and the mixture has just begun to melt around the wall of the erucible_
Cool the fusion mixture und pInee the crucible and contents in a 400-00.
bcaker. Add 150 ce. of hot distilled water and stir until the contents of the
crucible are completely dissolved."

Dried thyroid gland contains iodine in organic combination.

The organic matter is oxidized by a fusion mixture containing
alkali carbonates, which serve to fix the liberated iodine in the
form of iodide, and potassium nitrate, which acts as an oxidizing
agent. The carbonates used in the fusion mixture must be
anhydrous to prevent loss of material by decrepitation. A layer
of fusion mixture is spread over the surface of the mixture in the
crucible as a precaution against the loss of iodine. When thiH
mixture is heated a number of reactions undoubtedly take place,
but the only ones which pertain to the assay for iodine may be
regarded as:
2RI + R2C03~2KI + CO + H 0 2 2

RI + 3KN03~KI03 + 3KN0 2

In the above reactions it is assumed that all of the oxygen set

free when potassium nitrate is heated is not utilized in the oxida-
176 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

tion of the organic matter present but that some of it reacts with
K1 to form K10 3 After carbonization is complete and the
crucible has cooled, the residue is treated with distilled water
which dissolves the iodide, iodate, carbonates, and nitrite.
2. "Transfer the solution to a 500-cc. Erlenmeyer flask and rinse the
beaker arid crucible with four, lO-c(~. portions of hot distilled water, !1dding
the rinsirJgs to the solution in the flask. Cool the solution to 15C. and
add 50 ce. of freshly prepared chloriul1tcd soda T.S. Cautiously add 60 ee.
of dilute phosphoric acid (madc by mixing equal volumes of phosphoric arid
and distilled water), place the flask on a hot plate and boil the solution until
a strip of fllter paper, moistened with starch-potassium iodide T.S., does not
become blue when held in the vapor in the mouth of the flask. The final
volume of solution in the flask must be about 175 ce., and distilled water
must be ILdded, if necessnry, during the boililJg to maintain this volume."

Solution is best effected by treating the residue with successive

portions of warm distilled water. The combined solution is
treated with f;olution of chlorinated soda which functions as an
oxidizing agent to convert allY K1 present in the solution into
JUOs. The oxidation with sodium hypochlorite N aOOI is carried
out in solution acidulated with phosphorin .acid. The reactions,
although more complex, may be represented as:
2NaOCI + HsPOc-~Na2HP04 + 2HOOI
K1 + 3HOCl--'>K10 a + 3HOI
HOI + HOCl--'>HOH + Ch
Excess phosphoric acid is added to liberate all of the combined
chlorine present in the NaOOl. The solution is boiled to drive
off all free chlorine, since chlorine if present would liberate iodine
from KL
3. "Cool the solution to about 250. and add 10 cc. of a freshly prepared
aqueous solution of potassium iodide (1 in 100). Titrate the liberated
iodine with two-hundredth-normal sodiulll thiosulfate, adding 3 cc. of
arrowroot starch T.S. as indicator just befOl'e the end of the titration. Con-
duct a blank test with the same quantities of the same reagents, omitting
oIlly the Thyroid, and fusing as directed, and subtract the volume of two-
hundredth-normal sodium thiosulfate consumed froIn that consumed by the
Thyroid. Each cubic centimeter of the corrected volume of two-hundredth-
IwrIllal sodium thiosulfate is equivalent to 0.0001058 Gm. of 1."

When the cool solution is treated with K1 in acid solution,

iodine is set free quantitatively by the iodate:
 ()XIDA fl'JON AND lUmUC'l'lON 177

2KI + HaPO" K 2HP0 4 + 2HI

6HI + EI03~KI + 3H 0 + 3I 2 2

The liberated iodine is titrated with 0.005 N NU2S203:

Since each mole of KIO. obtained from the thyroid liberates

6 Gm.-atoms of iodine from KI, each aubie centimeter of 0.005
N N 0,2S2O3 IS . 1en t to 6 X 200
eqmva 126.92
X 1,000 00001058
=. . Grn.

iodine derived from thyroid glands.

Thyroxin and thymol iodide are assayed by modifications of
the above procedure.

Questions and Problems

1. What purpose is served by the alkali carhonates in the fusion mixture?
2. How does the ~ol\ltioll of ehlorilHlied sOlb function as nn oxidizing
agent?
3. Why must all of the liberated ehlorine be expelled previoull to the
!lddit.ion of ,In? Indicate by nn equation whal, would occur if the chlorine
was not driven off.
4. When KI0 3 is reduced by HI, what change of eharge does the iodine
combined as iodate undergo? Does the chang!) of ch[lrgc eorrespond with
the change of valence?
o. How may the USI! of 0.005 N Na 2S 20" ill place of 0.1 N NU 2S203 com-
monly used, minimize error in the measurement. of "olume?

TAllLE XXIV.-OFFICIAL SUBs'rANeES ASSAYED FOR IODINE BY CONVERSION"

OF THE IODINE TO IODA1'E AND TITRA'l'lON OF THE IODINE LIBERATED
FROM KI WITH STANDARD Na"s203

Amount
Nor- Equiva-
used, Official require-
Subst.ance mality of lent of
GIll. or mont, per cent
NU 2S20, 1 ec.
ec.
--- ----
U.S.P.
Calcium iodobehenate .. 0.5 0,1 0.002115 I2 = 23.5
Thymol iodide ... , ... , . 0,25 0.1 0.002115 h == 43
Thyroid ............... 1,0 0.005 0.0001058 12 == 0.17 to 0.23
Thyroxin.,., .... , ... , , 0.02 0.005 0.0001058 1I2 = 64
i ' I
178 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Exercise 59

Object.-Assay of Spirit of Ethyl nitrite.

Materials Required.-l0 ce. of spirit of ethyl nitrite (of known specific
gravity).
10 Gm. of potassiuIll iodide.
Cylinder of carbon dioxide.
10 cc. of dilute hydrochloric [Icid (1 in 2).
Aldehyde-free alcohol.
0.1 N sodium thiosulfate.
Apparatus.-"Fit a 300-cc. Erlenmeyer flask with a 2-hole mbber stopper.
Through one hole pass an aeration tube leading to the bottom of the flask and
tapered to an internal diameter of about 1 mm. at the lower end. Through
the other hole pass a glass tube, of at least 6 mm. internal diameter, which
will extend about 1 cm. above and below the stopper. Connect the aeration
tube with a cylinder or generator of air-free carbon dioxide."
Procedure.-1. "Place 10 Gm. of potassium iodide in the flask and add
40 ce. of boiling, deaerated distilled water. Insert the stopper in the flask
and pass a stream of carbon dioxide through the flask at a rate of 5 bubbles
per second until the solution has cooled to room temperature. Add 10 ce.
of dilute hydrochloric acid (1 in 2) and continue the stream of carbon
dioxide for at least three minutes. If any iodine is liberated, as shown by
the appearance of a yellow color, cautiously add tenth-normal sodium
thiosulfate from a burette, through the outlet tube, until the color is just
discharged. "

The water can be deaerated in the flask by boiling it and passing

a stream of carbon dioxide gas through it. Oxygen must be
excluded from the apparatus at all times during the assay since
it would react with the HI formed in the acid solution to liberate
iodine,

If iodine is liberated, the solution becomes slightly yellow in

color indicating that the solution was not completely deaerated.
2. "Decrease the flow of carbon dioxide to about 2 bubbles per second,
and add the directed volume of the nitrite solution with a transfer pipette,
passing the pipette through the outlet tube until the tip is just. above the
surface of the potassium iodide solution. Touch the tip of the pipette to
the outlet tube to remove adhering nitrite solution, then rinse the outlet
tube with a fine jet of aldehyde-free alcohol from a wash bottle. At once
titrate the liberated iodine with tenth-normal sodium thiosulfate, introducing
the tip of the burette through the outlet tube."
 OXIDATION AND REDUCTION 179
When the sample containing nitrite is introduced into the flask
nitrous acid is formed, and it reacts with the HI liberating iodine,
C 2H 5 0NO + HOH ~C2H50H HN0 2 +
75.05
2HN0 2 + 2HI~2HOH + 2NO + I2
The liberated iodine is titrated with 0.1 N Na 2SzOa in the usual
way.
Questions and Problems
1. Write equatiolls represrmting the n'uetions that oecul' in tlw Il~say of
amyl nitrite and spirit of glyceryl trinitrate.
2. Why must air be excluded during the nssuy of spirit of ethyl nitrite?
3. Ten cubic centimeters of spirit of ethyl nitrite (specific gravity 0.820)
required 30 ce. of a, solution of Na.S.03, each cubic centimeter of whieh i~
known to be equivalent to 0.01420 Gm. of ioclinc. Calculate the per cent
of ethyl nitrite in the sample.
TABLE XXV.-IODOMETRIC NITRITE ASSAYS

Equiva-
Amollnt lent of
Official requirement,
Substance used, 1 ce. of
per cent
Gm. 0.1 N
Na 2S20
.- --~

U.S.P.
Amyl nitrite ............ 3.5 0.01171 C,RtlONO = SO
Ethylllitrite, spirit of .... 10 0.00750/) C2H,ONO = 3.5 to 4.5
Glycel'yl tl'initrate, spirit
of .................. 1 25 0.0113 C,R,(ON0 2), = 1 to 1.1

TABLE XXVI.-OTHEU O.'FICIAL SUBSTANCES ASSAYED BY TITRATION OF

THE IODINE LIBEHATED FROM KI WITH 0.1 N NR 2S20,

Equiva-
Amount
lent of
used, Official requirement,
Substance 1 eo. of
Om. or per cent
0.1 N
ce.
Na 2S20,

U.S.P.
Chromium trioxide .......... 1 0.003334 CrO, = 95
Cupric sulfate ............... 1 0.01596 CUS04 = 63 to 66.S
PotaRsium bromate, R ....... 0.3 0.002784 KBrO. = 99.8

R. = reagent.
180 QUANTI'l'ATIVE PHilRMACEUTICAL CHEMISTRY

OXIDATION -REDUCTION METHODS WITH 0.1 N BROMINE

In the assay of compounds such as aniline, phenol, and resor-

cinol, bromine is employed 'as an oxidizing agent in place of iodine
since it forms tribrom derivatives quantitatively which are
insoluble in water in the presence of bromine in slight exc~ss.
The volumetric solution does not contain bromine as such, but
it contains potassium bromide and potassium bromate. Mix-
tures of these salts in solution liberate bromine when treated with
an acid.
Exercise 60
Object.-To Prepare and Standardize 0.1 N Bromine, 1{oppe-
scharr's Solution.
Materials Required.-3 Gm. of potassium bromate.
25 GIll. of pOkcssium bromide.
S ce. of hydroel1lol'ic acid.
S ce. of potassium iodide T.S .. (1fj.5 Gm. leI in 100 ce.).
0.1 N sodium thiosulfate.
Starch indicator solution.
Procedure.-Dissolve 3 Gm. of pbbl11sium bromatc and 25 GIll. of potas-
sium bromide in sufficient distilled water to make 1,000 ce. and determine its
exact normali ty as follows:
"Measure aC(Jurately from a buret.to about 25 ec. of the solution into a
500-ee. glass-stoppered IhlHk and dilute with 120 co. of distilled water. Add
5 ce. of hydrochloric acid, stopper the flask, and shake it gently. Then add
5 cc. of potassiull1 iodide T.S., re-stopper, shako the mixture, allow it to
stand for fiv(] minutes, and titrate the liberated iodine with tenth-normal
sodium thiosulfate, using starch T.S. as the indicator. C!llculate the
normality and, if desired, adjust exactly to tenth-normal."

When the solution of potassium bromide and potassium bro-

mate is acidulated with hydrochloric acid, bromine is set free:
51{Br + KBrOa + 6HCl--6KCI + 3Br2 + 3H 20
and the liberated bromine replaces the iodine of KI:
21{I + Br2--2KBr + 12
The flask should be tightly stoppered after the addition of the
acid to prevent the escape of bromine vapors.
The free iodine is titrated with 0.1 N sodium thiosulfate to
standardize the solution:
12 + 2Na 2S20a--t2NaI + Na2240~
 OXIDA'l'ION A.ND. REDUCTION 181

Since 1 mule of NaZS20S is equivalent tu 1 Gm.-atom of iodine,

and 1 Gm.-atom of iodine is equivalent to 1 Gm.-atom of bro-
mine, each cubic ccntimeter of 0.1 N Na~S203 is equivalent to
126.92 0012692 G . d' d 79.92
10 X 1,000 = . m.lO me an to 10 X 1,000 = (}.007992
Om. bromine.
If 20 CC. of 0.1 N bromine require 25 CC. of 0.1 N sodium thio-
sulfate, the solution ,vould be 25 ~O 0.1 := 0.1250 normal. Each
20 cc. of the bromine solution may be diluted to 25 cc. to make
an exactly 0.1 N solution, or the solution of known normality may
be employed.
Exercise 61
Object.-Assay of Phenol.
Materials Required.-1.5 Om. of phenol.
30 ce. of 0.1 N bromine solution.
10m. of potassiulll iodide.
1 cc. of chloroform.
0.1 N sodium thiosulfate.
Starch indicator solution.
Procedure.-l, Dissolve about 1.5 Om. of phenol, accurately weighed,
in sufficient distilled water to make 1,000 ce. Transfer an aliquot portion of
this solution, contltilling from 0.038 to 0.041 Gm. of phenol, to a 500 ec.
glass-stoppered flask hnving a long, narrow neck, add 30 cc. of 0.1 N bro-
mine, then 5 ce. of hydrochloric acid, and immediately insert the stopper.
Shake the flask repc!ttedly during P'2 hr., allow it to stnnd for 15 min.,
remove the stopper just sufficiently to introduce quickly 5 cc. of an aqueous
solut.ion of potassium iodide (1 in 5), being careful that no bromine vapor
escapes, and at once stopper the flask.

When the HCI is added, bromine is set free:

5KBr + KBrOa + 6HCl~6KCl + 3Br2 + 3H 0. 2

The bromine reacts with phenol to form a white crystalline

precipitate of tribromphenol and hydrobromic acid:

CoH.OH + 3Br2~C6H2Br30H + 3RBr

94.08
The flask should be tightly stoppered to prevent the escape of the
volatile bromine and Ilhaken gently during 30 min. to mix the
182 QUANTI'l'ATIYE PHARMACEUTICAL CHEMISTRY

eontents thoroughly and allow the oxidation of phenol to tri-

bromphenol to go to completion.
2. Shake the flask thoroughly, remove the stopper, and rinse it and the
Ileck of the Hask with a little dist,illed water, so, that the washings may flow
into the flasli., then fidd 1 ec. of ehlorofol'm, shake the mixture well, and
titrate with 0.1 N sodium thiosulfate, using starch T.S. as indicator. Each
cubic centimeter of 0.1 N bromine corresponds to 0.001568 Gm. of C.H.OH.

After thc addition of potassium iodide, 1 cc. of chloroform is

added to dissolve the precipitated tribromphenol which would
otherwise interfere with the clear observation of the end point,
particularly in the assay of old, colored phenol. The excess
bromine liberates iodine in accordance with the following
equation:
2KI + Br2->2KBr + 12
'The free iodine reacts with sodium thiosulfat.e:
12 + Na S20a-Jo2NaI + Na S 00
2 2 4

'The number of cubic centimet.ers of 0.1 N bromine added less

the number of cubic centimeters of 0.1 N Na 2S 20a required in the
titration gives the number of cubic centimeters of 0.1 N bromine
consumed in the reaction with phenol.
Since each mole of phenol oxidized to tribromphenol requires
6 Gm.-atoms of bromine, each cubic centimeter of 0.1 N bromine
consumec1 IS . Ient t 0 6 X 10
. eqUlva 94.08
X 1,000 = 0.00 11::68
D
Gm.

C6HDOH.
Liquefied phenol and resorcinol are assayed by the same
method. Acetanilid in tablets of acetanilid is determined by a
special method which is based upon the formation of trihrom-
aniline during direct titration with a standard bromine solution.
Questions and Problems
1. What substance is the oxidizing agent in Koppescharr's solution?
2. Explain the derivation of the equivalent weight of KBrO, when it is
used as an oxidizing figent in accordance with the change of charge which
the bromine atom undergoes.
3. How much pure KBr and pure KBrOa must be present in a solution .
which when treated with an acid will liberate an amount of bromine equiva-
lent to 100 cc. of 0.1 N iodine solution?
 OXIDATION A.ND REDUCTION 183
4. Calculate the weight of phenol and resorciuol, re,'ipectiYely, equiyalent
to 25 cc. of 0.1220 N bromine?
6. Aniline forms a tribromaniline when treated with bromine 3.na]ogoU;-f
to tribromphenol. Write equations for the reactions that occur ill the assay
of acetanilid in t.ablet.s of ilcetanilid..
6. Upon standardizing a solution of potassiuIll bromide-bromate against
acetanilid (see assay of tablets of acetanilid), 1 ce. of the solution was found
to be equivulellt to 0.008240 GIll. of acetanilid. A 0.2856 Gill. Ilumple of
powdered l1ect8!nilid tablets required 21.30 cc" of the brOlnide-hrOlUlLte solu-
tiOIl. If 20 tablets weighed 6.1244 GllJ., calculate the avemge amount of
acetanilid per tablet.
Exercise 62
Object.-Assay of anlmonium hypophoflphite.
Materials Required.-0.15 Gm. of amlllonium hypophosphite.
0.1 N bromine.
20 ee. of diluted sulfuric acid.
2 Gm. of potassium iodide.
0.1 N sodiulll thio~ulfate.
Procedure.-" Accurately weigh about 0.12 Gm. of the salt, dried over
sulfuric acid for 24 hours, and dissolve it in sufficient water to makc 100 ee.
TranlSfer 50 ee. of the solution to a 250-cc. glass-stoppered flask, a.dt! 50 ec.
of tcnth-normal bromine solution and 20 ce. of diluted sulfuric acid, stopper
the flask, shake well and allow to stand for three hours. Add 2 Gm. of
potassiUlIl iodide, dissolved in 10 ce. of recently boiled distilled water,
shnke the flask, and titrate the Hbcrat,ed iodine with tenth-normal sodium
thiosulfate, using starch T.S. as the indicator. Each cubic centimeter of
tenth-normal bromine is equivalent to 0.002077 Gm. of NH4PH~02'"
The hypophosphorous acid} formed when H 2S0 4 is added to
the solution of NH 4PH 20 2l is oxidized quantitatively to phos-
phoric acid upon standing in contact with the bromine solution:
2NH 4PH 2 0 2 + H SO r
2 -)o(NH 4)2S04 + 2HPH 0 2 2
2(83.07)
H HO
/
HO-P=O + 2Br2 + 2H20~HO-P=O + 4HBr '"
66.04
'" H
4(79.916)
HO/
98.04
Two of the hydrogen atoms in HP~02 carry negative charges
and are oxidized by the Br2 to positive hydrogen. The phos-
phorus undergoes no change of charge. The ionic equation
representing the reaction that occurs may be written:
184 QUAN'l'I'l'A'i'lVE PHARMACEUTICAL CHEMISTRY

2H- + 2Br2----?2Hi- + 4Bc

2( -) + zero = 2( +) + 4( - )
From the reactions, it is apparent that each cubic centimeter of
83.07
0.1 N bromine consumed is equivalent to
4 X 10 X 1,000
=

Questions and Prol>lems

1. How is ferric hypophosphite assayed? (See N.F. VL)
2. Derive the factor for total hypophosphites equivalent to 1 ee. of
0.1 N bromine for a mixture containing 50 Gm. of Ca(PII,02h 20 Gm. of
NaPH,02, and 30 Gm. of KPI-I 20, assuming 100 per cent purity for each
of the salts.
3. Give examples of a number of official substances other than those listed
in the appended table to which this method of assay might be applied .
. TABLE XXVII.-OFFICIAL SUBSTANCES ASSAYED BY TITRATING WITH 0.1 N
SODIUM THIOSULFATE 'rHE IODINE I,IBEHA'l'ED FROM POTASSIUM IODIDE
BY EXCESS 0.1 N BROMINE

EqlliVl1-
Amonnt lent of
Substance used, 1 ce. Official requirement, per cent
grams 0.1 N
bromine

U.S.P.
Phenol. .............. 1.5 0.001568 C OH 60H = 98
Phenol, liquefied ....... 1.5 0.001568 C,H 60H = 88
Phenol ointment ....... 2 0.001568 C OH 60H = 1.8 to 2.2
Resorcinol. ........... 1.5 0.001834 C OH 4 (OH)2 = 99.5
N.F.
Ammonium hypophos-
phite. " ............ 0.12 0.002077 NH.PH,O, = 97.5
Calcium hypophosphite 0.12 0.002127 Ca(PH 20,h = 98
Manganese hypophos-
phite ............... 0.12 0.002538 Mn(PH 20')2.H 20 "" 97
Potassium }lypophos-
phite ............... 0.12 0.002604 KPH 20 2 = 98
Resorcinol, mild paste
of ................. 1 0.001834 C,H 1 (OH)2 = 9.5 to 10.5
Resorcinol, strong p!1ste
of .... " ........... 1 0.001834 C.H.(OH)2 = 19 to 21
Sodium hypophosphite 0.12 0.002651 NaPH,02.H,O = 98
Sodium salicylate in
caffeine with sodium
salicylate ........... 2 0.002668 C 6H . OH.COONa '" 48 to 52
 OXlDL1TION AND REDUCTION 185

OXIDATION-REDUCTION METHODS WITH POTASSIUM IODATE

Potassium iodate may be used as the oxidizing agent in the

assay of a number of substances sllch as iodides, arscnites, and
other reducing agents. The method depends upon the formation
of iodine mono chloride in strong hydrochloric acid solution.

Exercise 63

Object.-To Prepare 200 CC. of 0.05 Molar Potassium Iodate.

Materials Required.-Ahout 2.2 Grn. of potassiulll iodate.
Procedure.-Dry 2.2 GIn. of pot,llssium iodate to constant weight at
110 e. Dissolve 2.1402 Om. of the dried salt in sufficient distilled water to
0

make exactly 200 ce. at 25C. The resulting solution should be exactly
0.05 molar.

The potassium iodate is dl'ied to constant weight at lloac.

to insure its freedom from moisture. Since potassium iodate is a
very stable salt and can be obtained in a very pure condition,
standard solutions of this reagent ean be prepared by dissolving
the calculated 'weight of the Halt in distilled water and diluting
to the proper volume.
Standard iodate solutions of known molarity rather than those
of known normality are usnally employed because the normality
varies depending on the nature of the reaction, e.g.,
1. Potassium iodate reactH with iodine in the presence of 12 per
cent or more of HCI as follows:
212 + RIO. + BHCI-KCI + 5IC} + 3H~O
and a normal solution of KIO a with respect to iodine should
contain ;..;1 mole.
2. Potassium iodate reacts with KI in the presence of 12 per
cent HCI as follows:
2KI + RIO. + 6HCI-3KCl + 3ICI + 3H 0 2

and a normal solution of RIGa with respect to KI should contain

H mole.
3. Potassiulll iodate reacts with As 2G a in acid solution as
follows:
186 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

and a normal solution of KIO a with respect to As 2 0 a should

contain 7:1 mole.
Exercise 64
Object.-Assay of Potassium Iodide.
Materials Required.-0.5 Gm. potassium iodide.
35 cc. of hydrochloric acid.
5 cc. of chloroform.
0.05 M potassium iodate.
Procedure.-" Dry about 0.5 Gm. of Potassium Iodide to constant weight
at 100C., weigh accurately, dissolve it in ahout 10 ec. of distilled witter, !Ldcl
35 cc. of hydrochloric acid and 5 cc. of chloroform. Titrate the mixture with
twcntieth-molar potassium iodate until the purple color of iodine disappears
from the chloroform. The last portions of the iodate solution must be
added in drops, the mixtlll'e being agitated vigorously and continuously.
After the chloroform has been docolorized, allow the mixture to stand for five
minutes. If the chloroform develops a purple color, the mixture Ilhould be
titrated further with the iodnte solution. Each (mbic centimeter of twenti-
eth-molar potassium iodate is equivalent to 0.01660 Gill. of K1."
PotassiUlll iodate is quantitatively reduced tQ iodine chloride
in strong HCI solution. Iodine is formed intermediately but at
the equivalent point, in the presence of sufficient Hel, it is com-
pletely converted into leI. The titration mixture should contain
at least 12 pel' cent of Hel at the end point to prevent the hydrol-
ysis of ICI. In solutions slightly acidified with HCI, the equation
representing the reaction is:
5KI + KI0 3 + 6HCl~6KCI + 31 2 + 3H 2 0
When sufficient HCl is present in the reaction mixture, the equa-
tion representing the reaction is:
2KI + KI0 3 + 6HCl~3KCI + 3ICl + 3H 0 2
2(166.02)
The change of charge in the latter equation is as follows:
21- + IH~3I+
2(-) + 5(+) = 3(+)
Each atom of iodine combined as iodide loses 2 electrons and each
atom of iodine combined as iodate gains 4: electrons. Each cc. of
0.05 M potassillm iodate is equivalent, therefore, to 2(166.02) =
20 X 1,000
0.01660 Gm. of KI.
 OXIDATION .AND REDUCTION 187

Chloroform is added to the titration mixture to enable the casy

and accurate determination of the end point. Binee all of the
iodine which remains during the latter part of the titration i"
collected in the chloroform layer, the disappearance of the violet
iodine color is readily observable. Vigorous shaking of the
titration mixture is necessary as the cnd point of titration if;
approached to mix intimately the immiscible solwuts so that tho
iodine in the ehloroforllllayer will be brought ill contaet with the
iodate.
Questions and Problems
1. Why should the i itration of i(]ditIes with iocllltn be (,llrried out in
strongly acid solution?
2. What is t.he function of the ehloroforlll ill the llS:;llY of iodides with
potassium iodate?
3. Write equations representing the change of ehargp t hat oet:ur~ whon
iodate is added to a solut,ioll of iodide which is slightly add.
4. 40 tnblets of red mercuric ioclido labeled" Ii 5 gmin tablets of mercurie
iodide" weighed 12.4220 Gm. A 10.0000 Gm. salJlple of the powdered
tablds required 11.45 ce. of 0.02 M KIO,. C:tlclliate tllP amount of Hgh
per tablet. Calculate the per cent devi:Ltion from the labeled amount.
5. AsccrLnin the method of ltssay of iodine ointrncnt, in the U.S.P. Ex-
plain the procc(lure.
TABU] xxvrII.-Ol'FICIAL SUBS'rANGES ASSAYED BY TITRATION Wl'l"H KIO

Substunce
Anl()Unt
used, .I EquiV"ll-
1Il0hmt.y I t f Offwiu ll'cq uirerl1ent,
Om. or of 100, ~ncc~ })er cent
ce.
_-_ ---~

U.S.P.
Chiniofon, powder ............. 0.3 0.05 0.01269 ! = 26.5 to 28.0
I<;>dophthulein, soluble .......... 0.3 0.05 O. 01269 I = 61 to 62
Oil. iodized ................... 0.35 0.05 O.012fl9 I = 39 to 41
Potassium iodide. 4 , 0.5 0.05 0.01060 In = 90
Sodium iodide ............ 0.5 0.05 0.01490 Nul = 99
N.F.
Arsenic trioxide, tablets of ...... 0.06" 0.02 0.003956 "\s,O, = 92.5 to 107.5'
11ercuric iodide, ted .... ........ 0.2 0.05 0.02272 HgI, = 99
Mercuric iodide, rod, tablets of. . 0.13" 0.02 0.009089 Hgh = 91 to 109'
Mercurous iodide, yellow, tl1blets
of. ......... " ....... , ..... 0.1- 0.02 0.008734 HgI = 91 to 10gb
PotaBsium iodide, solution of .. 5 0.05 0.01660 KI = 97 to 103lV/V
Potassium iodide, tablets of, ..... 0.1" 0.02 0.OOtlfl40 KI = 1l2.5 to 107.510
Sodium iodide, ampuls of ........ 0.5 . 0.05 0.01490 Na! = 92 to 105'

"Amount of ingredient sought.

b Per cent of the labeled amount,.,
 CHAP'l'ER X

GASOMETRIC METHODS
The assay processes which involve the volumetric measure-
ment of gases are of two types, namely:
1. The purity of a gas is determined by absorbing one of the
components of a gaseous mixture by means of a suitable absorbing
agent and measuring the quantity of gas absorbed by the change
in volume.
2. The purity of a given substance is determined by measuring
the quantity of gas which it may be made to evolve in a given
chemical reactioll.
Theory.-'-According to the kinetic theory of gases, the mole-
cules of a gas are in constant motion. When a gas is enclosed in a
vessel, it exerts a definite pressure due to the combined effects of
the impacts of the moving molecules of gas upon the walls of
the containing vessel. The magnitude of the pressure exerted
depends on the number of molecules which strike a given surface
in a given period of time. The number of molecules which strike
upon the surface in a given period of time is dependent upon the
number of molecules present and upon the velocity of the mole-
cules. The velocity of the molecules varies with the temperature
of the gas; the higher the temperature the greater the velocity of
the molecules, and the lower the temperature the less the velocity
of the molecules. If the temperature and pressure of a gas remain
constant, the volume of the gas will depend on the number of
molecules present.
The official gasometric determinations pertain only to sub-
stances which behave as "perfect" gases, that is, gases in which
there is no dissociation or association of the molecules, and which,
therefore, obey the gas laws. Consequently, a definite volume
of such a gas measured under laboratory conditions at a known
temperature and pI'essure provides the data required to calculate
the weight or volume of the gas present under standard condi-
188
 (iflS0METRIC METHODS 189

tions, as will be shown subsequently. The term standard condi-

tions is used to designate a temperature of OC. and a preSFiure
equal to that exerted by a column of mercury 760 mm. in height.
Whenever the volume of a gas is given, it is nnderstood that t.he
gas oecupies the specific volume when meu:sured at O"C. aud
at a barometric preSSlll'e of 760 !Iun. unless otherwit'{(J "tated.
Standrirel conditions should not be confuRed with the expression
normal conditions which has l'cferenee to the meftsnrement of
g11,':leS at 25C. and 760 mm. pressure.
It has been found by experiment that 22.4 l. of oxygen mnas-
lll'ed at OC. under a pressure of 760 mIll. weighs 32 Gm. and thltt
the same volume of hydroge1lll1easured under standard conditiolls
weighs 2.016 Gm. Thetle facts are embodied in Avogadro's
hypothesis, which is: Equal volumes of gases measured under the
same conditions of temperature and pressure contain the same
number of molecules. In the case of perfect gase>;, this hypothesil:l
may be stated; Equivalent molccular weights of gascs confined
under identical conditions of temperature and prcssure occupy the
same volume. Thus, 2.016 Gm. of hydrogen, 32 Gm. of oxygen,
30.01 Gm. of nitrie oxide, or 14.008 Om. of nitrogen will oeeupy
22.4 l. when measured at (lO. and 760 mm. preRsure. According
to Boyle's law: The volume of a gas is 'inversely proportional to
the pressure upon it if the temperature remains constant. Doubling
of the pressure would diminish the volume one-half, and, con-
versely, diminishing the pressure one-half would double the
volume. The prodU(~t of the preS~lUre and volume is therefore
constant. Boyle's law may be stated in algebraie form as follows;

or Yl = P2 = K
Y2 1\
where P 1 and V 1 are the pressure ami volume under one set of
conditions and P2 and V 2 are the pressure and volume under
another set of conditions, K being their constant product. The
magnitude of K depends upon the quantity of gas measured. If
P l and VI are used to represent the observed pressure and volume
of a gas, respectively, and P 2 is standard pressure (760 mm.),
the volume V 2 which the gas will occupy at P 2 may be calculated;
e.g., if a sample of gas measured at 25C. and 750 mm, pressure
occupies a volume of 50 ce., what volume will it occupy at 250.
190 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

and 760 mm. pressure? By making the proper substitutions

in the preceding equation,
50 760 50 X 750
VI = 750 and VI = 760 = 49.35 cc.

The increase of 10 mm. pressure on 50 cc. of gas measnrccl at

25C. is thus found to have caused a decrease in volume of 0.65 cc.
Charles' law may be expressed as follows: If the volume of a
definite mass of gas is kept constant, the pressure which it exerts
is directly proportional to the absolute temperat'U?'e at whz"ch it is
measured. This law may be stated in algebraic form as follows:
111 T1 or VI = V 21'1
V2 = T2 1'2
where VI and V 2 are the volumes of the gaR at the absolute tem-
peratures 1'1 and 1'2. Zero on the absolute temperature scale
corresponds to -273C. The absolute temperature at which
a given analysis is carried out is therefore found by adding 273
to the laboratory temperature expressed in degrees centigrade.
The above equation then becomes
VI 273 + tl v _ V2 X 273 + tl
or
V2 = 273 + t2 and 1 - 273 +
tz
where tl and t2 represent the respective temperatures in degrees
centigrade; e.g., if a sample of gas measurcs 50 cc. at 25C.,
what volume would it occupy at OC., the pressure remaining
constant? Upon substituting in the above equation, wc obtain
50 273 + 25 _ 50 X 273 _ ~
and V2 298 - 4;).81 cc.
V2 - 273 + 0 -

When both the temperature and pressure at which a gas is

measured deviate from the official requirement, the corrections
for each may be made in succession, as illustrated in the above
examples. It is more convenient, however, to combine the
respective equations which represent Boyle's law and Charles'
law and make the correction in a single calculation. The com-
bined laws may be expressed by the algebraic equation

111 = P 2V 2T 1
P11'2
 GASOMETRIC METHODS 191

The following example illustrates the use of thiR equation. - A gas

measures 40 ce. at 200. and 740 mm. pressure. 'What volume
will it occupy at 250. and 760 mm. pressure? By substituting
the propel' values in the foregoing equ!1tion, we obtain
T 74{) X 40 X 298
T1 = 760 X 293 = 39.7:) co.

Apparatus.-The volumetric measurement of gases in tIl('

official assay processes is made by the use of a gas burette of the

;1'
~ I:

,I'.:
,

.. '

1
!! 1:"':
'a- Q'!

,I,t!
FIG. I5.-Gas FIG. 16.-Simple gas pipette.
burette with level- (Courteay Fisher Scimlti/ic Com-
ling flask. (COU1'- pany.)
teay Fisher Scienti-
fic Company.)

type illustrated in Fig. 15. The burette should be of 100 cc.

capaeity and graduated in 0.2 cc. divisions. The burette may
1;>e enclosed in a water jacket, as illustrated, to permit the
control of temperature. The lower end of the gas burette is
attached to an op'cn leveling or equilibrium tube by meallS of ,:),
heavy walled rubber tube.
The absorptioll of the gases is accomplished in a gas pipette
similar to that shown in Fig. 16. The absorption bulb of the
192 (JUAN'l'I'l'ATIVl? PllARMAC'}j,'U'l'IC'AL CIIEMIS'['RY

pipette is provided with an opening at the bottom which can be

closed by means of a rubber stopper and through which solid
absorbing agents may be introduced.

Exercise 65
Object.-Assay of Carbon Dioxide.
Materials Required.-Carboll dioxide compressed in a metallic cylinder
which is provided with a reducing valve.
A gas burette with leveling tube.
A gas pipette.
About 2 lb. of mercury.
150 cc. of 50 per cent KOH solution.
Procedure.-l. "Place a sufficient quant,ity of mereury in lL IOO-ce. gas
burette or nitrometer, provided with a two-way stop-eock and a two-way
outlet, IInli properly connected wit,h a balancing tube. COllnect olle of the
intake tubes of' the nitrometer with a gas pipette of suitable eapaoity.
Place in the pipette about 125 cc. of 50 per cell t potassiulll hydroxide solu-
tion. Draw the liquid (frce from air bubbles) through the capillary opening,
connection and stop-cock opening in thc llitrometel' by reducing the pressure
in the nitrometer tube and opening the stop-cock controlling the connection
with the gas pipette. Then close the stop-cock.';

. The gas burette is filled with mercury by raising the leveling

flask containing the mercury until the stopnock opening is filled
with the mercury. The stopcock is then closed. The gas
pipette is filled with 50 per cent KOH so that the absorption bulb
and capillary tube are completely full and the expansion bulb
is about one-tenth full and is connected to one of the outlet tubes
of the gas burette, Upon opening the stopcock connecting the
pipet.te and t.he burette and lowering the leveling flask, the KOH
solution and any trapped air in the connection are drawn through
the stopcock opening. Any air bubbles drawn into the burette
may be driven out by opening the stopcock to the other outlet
and raising the leveling flask. .
2. "Having completely filled the nitrometer, the other stop-cock opening
and the other intake tube with mercury, draw int.o the nitrometer exactly
100 ce. of Carbon Dioxide, measured at atmospheric pressure and at 25C.,
by reducing the pressure in the tube."

The gas can be introduced directly into the burette from a

cylinder of the compressed gas if the latter is provided with a
reducing valve which can be carefully controlled. When the
 GASOMETRIC ME1'IlOD/3 193

gas is obtained in this manner, the connectioJl tubing between

the cylinder and the burette should be flushed free of air by
allowing a stream of the gas to flow through the tubing for a
minute or two. A volume of gas slightly in excess of 100 co.
is then admitted to the burette, and after it has aequired a
temperature of 25C., the excess gas is permitted to escape so
that a volume of 100 cc. remains.

3. "Close this stop-cock. Increase the pressure on the C:Lrhon Dioxide

in the nitrometer tube and open the stop-cock controlling the eOIlllcction
with the gas pipette. Forco the entirlJ volume of gC{S into tlJe pipette.
Clese the stop-cock and rock the pipette gently, proviLling frequent con bet.
of the liquid and gas. At the end of five minutes most of the gas will have
been absorbed by the liquid. At this time, to faeilit[Lte the ahsorptioll of
the last portion of the gas, draw some of the liquid htto the nitrometer tube
aud force the residual gas back upon the surface of the liquid in the gas
pipette. Again rock the pipette until no further diminution in the yolume
of gas occurs. Draw the residual gas, if any, into the nitrometer tube and
measure its volume at atmospheric pre~sure and at 25C.
"The volumo of gas remaining uudissolved should not exceed 1 ce., indi-
cating not less than 99 per cent by volullle of CO, in the gas tested."

The CO 2 is absorbed by the ROR forming K 2COsleaving other

gases which are not absorbed by l{OH as fPsidual gas. If thp
initial volume of the sample was exaetly 100 ce" the decrease
in volume is numerically equal to the per cent of CO 2 in the
sample.
All the official gasometric assays are performed by the absorp-
tion method. The procedures are designed primarily for use in
manufacturing laboratories for the control of the purity at the
time the gases are compressed into cylinders.
TABLE XXIX.-OFFlC'IAL GASOMETlUC ASSAYS

Official rllquire-
Substance AbsorptiOll reagent
ment, per cent

U.S.P.
Carbon dioxide. . . . .. Potassium hydroxide CO 2 =- 99
Ethylene. . . . . . . . . . .. Bromine CH 2 = CHa = 99
Nitrogen monoxide.. Water N 20 = 95
Oxygen. . . . . . . . . . . .. Copper with NH 4Cl and o "" 99
Nli.OH
194 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Although no assay process involving the measurement of

evolved gases is official, the procedure employed in this type
of assay is of such general application that every student of
pharmaceutical analysis should he familiar with the method.
The following exercise illustrates this method
D
of gasometric analysis.
Apparatus.-The volumetric measurement of
evolved gases may be made by the use of an
instrument known as a nitrometer. The nitrom-
eter of the type devised by Lunge (Fig. 17)
consists of a gas burette A of at least 50 cc.
E capacity, graduated into 0.1 cc., with a special
glass stopcock B at the top, to which is sealed a
glass cup D. The gas burette is connected with
an open leveling or equilibrium tube E by means
of a heavy-walled rubber tube F.
In the use of the simple Lunge apparatus
described above, both tubes of the instrument are
__..........:F partly filled with some liquid in which the gas to
be measured is insoluble; generally mercury is
FIG. 17.-Lunge employed but in some cases a saturated i:lalt
nitrometer.
solution is used. The latter may be used in the
measurement of the nitric oxide liberated from ethyl nitritc or
amyl nitrite.
When mercury is used, enough of it is poured into the equilib-
rium tube to fill the burette and leave a slight excess. The
stopcock is turned to connect the gas burette with the glass cup,
and the equilibrium tube is raised until the mercury level reaches
the bottom of the cup to expel all of the air from the burette
and stopcock bore, and the stopcock is closed. The sample to
be assayed, a liquid or a solid in solution, is weighed and poured
into the cup from a weighing bottle or weight pipette. Upon
opening the stopcock and lowering the equilibrium tube slightly,
the sample is drawn into the burette. Care mURt be exercised to
prevent thc entrance of air into the passage of the stopcock. The
cup is rinsed two or three times with 3 cc. portions of the reagents
used to liberate the gas, each being drawn into the burette al:]
described above. With the stopcock closed, the equilibrium
tube is lowered sufficiently to cause a slight diminution of the
 GASOMETRIC METHODS 195

pressure in the burette, and the latter is shaken vigorously for

about 5 min. to bring the reagents and sample into intimate
contact. When no more gas is evolved on further shaking, the
burette is allowed to stand about 30 min. to permit its contents
to acquire room temperature. The volume is then meflsured at
atmospheric pressure. Since there is a column of impure reac-
tion mixture above the mercury in the burette, SOllle proeeclurc
must be employed to adjust the level of the equilibrium tube so
that the weight of the column of mercury in it will just equal the
weight of the columll of mercury and reaction mixture in the
burette. If the reagent mixture has a speeific gravity of about
1.3, as it doeR in the assay for et.hyl nitrite, the equilibrium
tube is set 011 the assumption that lllllll. of morellI'y will bala.nce
about 10 mm. of the reaction mixture. About 5 ceo of dilute
sulfuric acid is then poured into the cup and the stopcock is
carefully opened to it. The tendency to draw in or expel gas is
noted and the equilibrium tube is adjusted aecordingly. This
tediolls leveling procedure is obviated ill the assay by the use of
a saturated flolutioll of sodium chloride in the place of mercury.

Exercise 66

Object.-To Prepare and Test a Nitrometer.

Materials Required.-l gas burette.
I equilibrium tube.
30 in. of nitrollleter tubing.
A satumted solution of sodium chloride.
Procedure.-Clcall tlw gas burette and equilibrium tnbe of the uitrometer
thoroughly, connect them by :1 piece of heavy-w!Llled rubber tubing about
3D in. long, grease the burette stopcock with a thin film of petrolatum, and
clamp the assembled instrument securely to a Slipport in It vertiCil'! position.
Open tbe stopc:o(:k of t.he burett!l and pour sllffieient slltl1rntcd solution of
sodium chloride into the equilibrium tube to fill the burette and the bore of
the stopcock completcly when the cquilibrium tube is raised. Carefully
adjust the lcvel of the equilihrium tube so that the stopcock passage is
completely filled with the salt solution without !tny of it being present ill thc
glass eup and close. the stopcock. Lower the equilibrium tube so that the
solution level in it corresponds approximl1tely with the 50 ce. grad\llltion
on thc bl\l'ctte, note the solution level in the burette, and after the instrument
has stood for 20 min. talce another reading. If the two readings do uot
correspond, the stopcock is leaky, If the leakage cannot be corrected by
the use of stopcock grease, another instrument should be employed.
1!}6 (JUANTI'l'A'l'IVE PJIAJlMACEU'l'ICAL CHEMISTRY

Exercise 67
Object.-Assay of Spirit of Ethyl Nitrite.
Materials Required.-l Lunge nitrometer.
1 10 cc. delivery pipette.
J 100 cc. volumetric flask.
40 C(~. of ~pirit, of ethyl nitrite.
0.5 GIll. of potassium bicarbonate.
(){) CIl. 01 1111l0ho1.
1.65 Grn. of potassium iodide.
5 ce. of diluted sulfuric acid.
Procedure.-l. Transfer about 40 cc. of spirit of ethyl nitrite, which
haR heen previously slllLken with 0.5 Gm. of powdered potassium bicarbon-
ate, to a tared, 100 cc. measuring flask, and weigh accurately. Add sutli-
cient alcohol to bring the volume to exactly 100 cc. and mix thoroughly.

In the presence of water, ethyl nitrite is partially hydrolyzed

with the formation of ethyl alcohol and nitrous acid:
C 2H 5N0 2 + H 0--7C H 0H + HN0
2 2 5 2

Since the nitrous acid, if present, would liberate oxide in thc

assay, it is removed by shaking the spirit with powdered pot.as-
sium biC:Lrbollate:

The potassium nitrite formed is practically insoluble in the spirit.

The sample is diluted with alcohol to 100 ce. after weighing, so
that an aliquot portion of the solution representing a definite
weight of the spirit may be measured into the nitrometer because
it is difficult to weigh and ~ramlf(~r a sample of the spirit directly
without the loss of some ethyl nitrit.e.
2. Int.roduce into a nitrometer exactly 10 cc. of the alcoholic solutioll,
follow by 10 cc. of potassium iodide T.S. and afterwardl5 by 5 cc. of diluted
sulfuric acid. Whcn the volume of gas has become COllstant (within 30 to
60 min.) note the volume of gas collected.

Adjust the nitrometer prepared for use in the preceding

exercise so that the level of the salt solution in the equilibrium
tube is about even with the 15 cc. graduation mark on the burette,
the latter as well as the stopcock passage being filled with the
.~alt solution. Fill a 10 cc. pipette with the diluted spirit exactly
to the graduation mark and transfer the liquid into the nitrometer
 GASOME'1'IUC ME1'llODS 197

cup. Open the stopcock at once to allow the solution to be drawn

into the burette llsing cure to prevent the entrance of air at the
same time. R.inse the cup at once with two .5 ce. portions of
potassium iodide solution (1.65 GIll. 1(1 in 10 ec. H 2 0), drawing
each portion into the burette by lowering the equilibrium tube,
if necessary, and then place about G cc. of diluted sulfurie aeid
in the cup and draw about 5 cc. of it into the burette in the same
manner. The excess acid remaining in the cup serves to prevent
the escape of the nitric oxide gas 'which begins to form as soon as
the first of the acid is introduced, and it prevents the (~ntr[Lllee of
air. After the stopcock has been tightly closed and the equilib-
rium tubc well clamped to the support, remove the IJUl'l't te
from the clamp, in dine it at an angle of about 45 deg., and shake
it vigorously for about 5 min. to mix the n~ngents thoroughly
with the solution of ethyl nitrite.
The sulfuric acid liberates hyciriodie acid from the potassium
iodide:
KI + H2S0~~KHS04 + HI
and the liberated hydrioc1ic add reacts with the f~thyl nitrite in
the spirit, setting nitric oxide gas free:

The entire reaction may be represented in one equation, as:

The nitric oxide gas formed being insoluble in saturated salt

solution rises to the top of the burette, and after it has acquired
room temperature as shown by the constancy of its volume when
two successive readings taken at 10-min. intervals agree, the
volume of gas libemted is recorded. When the final reading of
the volume of gas is taken, the liquids in the burette and equilib-
rium tube should be on the same level.
Since 1 Gm.-molecular weight of a gas at ODC. and 760 mm.
pressure occupies 22.4 l., 1 I. of a gas will contain the gram-
molecular weight divided by 22.4. The volume of 1 Gm.-molecu-
lar weight (30.01 Gm.) of nitric oxide gas measured under OC.
and 760 mm. pressure, is 22.3855 l. One liter of nitric oxide gas
will therefore contain 30.01/22.3855 = 1.3406 Gm., and 1 cc.
198 QUANTITATIVE pHARMACEUTICAL CHEMISTRY

of the gas will contain 0.00134:06 Gm. At 250. and 760 mm.
presfmre, 1 l. of the gas will contain 1.34:06 X 2732~ 25
1.2281 Gm., and each cubic centimeter of nitric oxide gas will
eontain 0.0012281 Gm. of NO.
The volume of gas being known and the temperature in degrees
centigrade and the barometric pressure in the laboratory having
been aKeertained, the ""veight of nitric oxide gas evolved by a
given weight of spirit of ethyillitrite may be calculated; e.g., an
tdiquot portion of an alcoholic solution of spirit of ethyl nitrite
equivalent to 4.0 Gm. of the spirit when assayed evolved 50 cc.
of uitric oxide measured at 21e. and 740 mm. pressure. What
percentage by weight of ethyl nitrite did the sample contain?
Substituting in the algebraic cquation which expresses the COffi-
binedlaws of Boyle and Charles,
V 1 = !:..iY2Tl
P!T!'
've obj,aln V = 740 X 50 X 298 = 4935
1 760 X 294 . cc.,
the volume corrected to 25C. and 760 mm. pressure. 49.35 X
0.0012281 = 0.06061 Gm. NO contained in 49.35 cc., at 250.
and 760 mm. pressure.
Since 1 mole, 75.05 Gm., of ethyl nitrite, C 2H 5N0 2, evolves 1
mole, 30.01 Gm., of nitric oxide, NO, each gram of nitric oxide is
the approximate equivalent of 75/30 = 2.5 Gm. ethyl nitrite,
and 0.06061 Gm. of NO is equivalent to 0.06061 X 2.5 = 0.1515
GIn. 02H 5N0 2
The percentage by weight of ethyl nitrite contained in the
samp1e IS there fare --4:0--
0.1515 X 100 =. 379 pel' cent. 0 alculate

the percentage of ethyl nitrite present in the sample of spirit

assayed.
Comment.-When a gas is measured above a liquid, the total
pressure observed is due to the vapor pressure of the liquid plus
the pressure of the gas. In the assay of spirit of ethyl nitrite,
the nitric oxide gas evolved is measured above a solution mixture
consisting primarily of water and alcohol., According to Dalton's
law, each gas in a mixture of gases exerts the same pressure as it
would if it were present alone. The vapor pressure of water
at 250. has been found to be equal to 23.55 mm. of mercury at
OC., and that of alcohol has been found to be equal to 5.94 mlll.
of mercury at OC. The pressure exerted by the nitric oxide gas
 GASOMETRIC METHODS 199

in the preceding example should, therefore, be corrected for the

vapor pressure of water and alcohol 11S follows: 740 - (23.55
+ 5.94) = 7l0.5mm. Unless this correction i" made, thc method
of assay measures the water and a1<:oho1 vapor;:; present as nitric
oxide.

Questions and Problems

1. Upon what factors is the pre~surc exerted by a gas dependent?
2. How do the "nornuLl conditions" eIllploy(~d ill the lIWtlRUl'l!IIJf'ut of
gases in the Pharmacopoeia differ from stuml:ml cllllditionR'?
3. How many grams of oxygen, hydrogen, and nit,rio oxiilf! gas, rc~pce
tively, will be contained in a volume of 1 l. Ilwilsured at ODC. ~lUd 700 mill.
pressure?
4. 60 ce. of nitric oxide gas is cooled at constant prC8Slll'e from 40 to 25C.
Calculate the volume of gas at the latter temperature.
6. A gas occupies a volullle of 40 ce. at 20C. and 705 nlln. pressUl'P.
What volume will it occupy under standard conditiOllH?
6. An aliquot portion equivalent, to 1.2 Gm. of spirit of ethyl nitritll
evolve8 48 ce. of nitric oxide llIe!lSllrcd at 25C. tlIId 742 nun. pf[~HB1Il'e.
Calculate the percentage by weight of ethyl niN'ite contained ill the sViril.
7. If the gas in problem (J was collected over a solution consisting primarily
of alcohol and water with vapor preSRures equal to 5.1I4 lInrl. and 23.55 llllll.
of mercury, respectively, what percentage of ethyl nitrite wonlLi the solution
contain?
8. From the results obtained in problemH 6 and 7 cakulate the pur cent
of error in the amount of ethyl nitrite containcd in the spirit [h,\!; is CaUH()[\
when the corrections for the vapor presimrcs of wate)r and nlcohllI aro not,
made.
9. Write equations represrmting the reactions that take phwe in the
absorption of the gllses ill tho oflicilll gllsomctric assays.
 PART II
PHYSICO-CHEMICAL METHODS USl:D IN
OFFICIAL PHARlVIACEUTICAL ANALYSIS
The analysis of many of the official substrLllces for the purpose
of identifying them and of determilling their purity or therapeu-
tic value is based on the measurcnwnt of certain physical und
chemical characteristics or constants, considered in conjunction
with the estimation of certain special components by assay, and
qualitative tests for various suhstances commonly prcsent as
impurities or added for the purpose of adulteration.
The qualitative methods employed in the determination of the
physical characteriHties or physical eonstullts of official substances
are frequently applied to materials of wiclely divergent. nature;
e.g., valuable criteria relative to the purity of volatile oils, sugars,
and alkaloids may be gained by the meu.'iurement of the optical
activity. The fundarnental principles involvecl in ascertaining
the physical eonstants of these :mbstances are the same regardless
of the nature of the substance, although slight modifications
of the technique employed are frequently necessary.
The Pharmacopoeia and National Formulury, as a rule, spPcify
that certain methods itnd itpparatuH shall be employed in the
determination of the phYHical constants of official substances.
In such cases, the methods are known as official methods and the
appamtus as official apparatus.
 CHAPTER XI

SOLUBILITY

The statements of the solubilities of official suhstautes un!

given primarily for the informatioll of physicialls and l1harmaei"ts
for use in the preparation and dispcu:;illg of medicilwH. TI1f'Y
should not always be regarded as exact phYi;ical eon,.;tants upon
which the identificatioll or the determination of the purity of a
substance may be based. Frequently, the official standards arp
descriptive terms to indicate the approximate solubility of sub-
stances, viz.: (1) It Very soluble," 1 part of the substanee is Holuble
in less than 1 part of the solvent; (2) "freely snluhl(~," 1 part of th!'
substance is soluble ill from 1 to 10 parts of solv('ut; (3) "solullln,"
1 part of the substance is soluble in from 10 to 30 parts of sulvent;
(4) "sparingly soluble," 1 part of tho substnnec is solublo in frulll
30 to 100 parts of solvent; (5) "slightly soluble," 1 part of the
substance is soluble in from 100 to 1,000 Pfuts of solvent; (i)
"very slightly soluble," 1 part of the sulmtullep is soluble in from
1,000 to 10,000 parts of solvent; and (7) 'ipractimlly insoluble,"
more than 10,000 parts of solvellt arc rC(luired to dissolve 1 part
of the substance.
There are nine possible classes of solutiolls, naIlwly: (1) a gas
in a gas; (2) a gas in a liquid; (3) a gaR in a solid; ('1) a liquifl
in a gas; (5) a liquid in a liquid; (6) a liquid in a solid; (7) (t
solid in a gas; (8) a solid in a liquid; and (9) a solid in a solid.
Although examples of all of these different type::; of solutions
are known, the only classes of pharmaetmtical importan('e are
solutions of gases in liquids; of liquids in liquids, and of soliclH
in liquids.
The official determination of the solubility of gases in water,
such as HCI and NH3; is generally ('ffected by chemieal titration
methods, while the solubility of liquids in liquids and of solids
in liquids is determined by physical methods.
203
204 QUAN7'ITA1'!VE PHARMACEUTICAL CHEMISTRY

Numerous factors affect the rate and the extent of the solu-
bility of a substance in a given solvent as follows:
1. The solubility of most of the official substances is increased
by a rise ill the temperature at which solution is effected. There
are numerous exceptions to this generality, however; e.g., gases,
ealcium salts, and ether are less soluble in hot than in cold water.
Since the solubility of substances varies markedly with slight
changes in temperature, it is very important that a constant
tcmperature be accurately maintained throughout a solubility
determination.
2. Substances in a fine state of division dissolve more rapidly
than large crystals or particles because the total surface area
exposed to the action of the solvent
is m.'1ch greater when the substance
is powdered.
3. The purity of the substance and
of the solvent must be assured in all
solubHity determinations, since slight
amounts of impurity in either may
cause considerable variation in the
results. In the official statements of
solubility, it is assumed that both the
substance and the solvent employed
conform to the official tests for purity.
4. The position of the solute in the
solvent affects the rate of solution.
If the solute is allowed to lie on the
FIG. IS.-Apparatus for sol-
ubility determinations. A As- bottom of the vessel, it becomes
sembled apparatus. B Stirrer. surrounded by a layer of concentrated
solution which prevents the access of fresh portions of the solvent
to the surface of the solute.
The determination of the solubility of a solid in a liquid neces-
sitates the preparation of a saturated solution. The production
of a saturated solution of this type may be carried out in .the
apparatus illustrated in Fig. 18. This apparatus consists of a
hard-glass test tube A of medium size into which the solvent and
solute are placed and stirred vigorously by means of a motor-
driven glass screw stirrer B. The stem of the stirrer passes
through a glass tube, inserted through a well-cleaned stopper by
 SOLUBiLiTY 205
which the soluhility tube is closed. The glass tube selected
should be of such :1 size that the stem of the :-:tirl'er jllst pass!';.;
through, the area, of contact being well lubrie/1ted with petro-
latum. When solubility cleterminations nre madC', the solubility
tube is fastened npright in a thermostat maintllilH~cl at a eOIl:':tant
temperature of 2.5C.
Exercise 68
Object.-To Determine the Sulubility of Boric Acid in Water
at 25C.
Materials Required.-Uj Om. of horic ~wid.
A solubility app:.Ll'[ltub auel t,hermo~tnt.
Procedure.-1. In the solubility tube, plnec: about 1.5 Om. of fhwly
powdered boric acid, 25 ec. of distilled water, [Illd after fitting it with the
stirrer, place it in the thermostat so that it is immersed in the water up to
the level of the stopper, and stir the ~olution vigorously for about 3 hr.

The boric acid is powdered finely because substances ill fine

particles dissolve more rapidly than do large crystals; subdivisioll
increases the surface area exposed to the aetioll of the solvent.
The solubility tube is il1l111Crspd in the thermostat bath, to
maintain a constant tcmperatmc throughout the mixture, since
the solubility of boric ncid varies appreciably with slight changes
in temperature. The temperature in the thermostat should not
vary by morc than O.lC., above or below 25C., during the
determination. The mixture is stirred continuously to bring
fresh portions of solvent in contact with the undissolyed. particles
of boric acid and insure equal cOlleentration throughout the
solutioll.
2. Stop the stirrer Illul reJt1ov() it from the tube, stopper the 1(1 tter secnrely,
allow the untlissoked crystals to settle, remove about 5 ce. of .the dear
Holution frolll the solubility tuhe by means of a pip(ltte, and weigh accurately.
Evaporate this portioJl of the solution to dryness on a water bath at 80C.
nnd finally to COllstant weight :1t 90C. in an elcetric or other sllit!lble
oven, and weigh the residue.

The weight of the boric acid contained in the residue, sub-

tracted from the weight of the solution analyzed, gives the ,,,eight
of the water in which the boric acid was dissolved.
8. Attach the stirrer and allow the stirring to continue for about one
hour. Again remove a 5 cc. portion of the solutioll, weigh it accurately,
206 QUAN1'1'l.'A'l.'IVE PHARMACEUTICAL CHEllHSTRY

and determine the amount of boric aciri contained in it in the same manner
as before.
If this result agrees with the former determination, it shows
that satnration of the solution is complete, but if the amount of
dissolved solid is greater in the second case, the stirring must be
continued until the concentration of tho solution becomes
constant.
4. Express the results as gra.ms of boric acid soluble in 100 Gm. of water.

If a 5.5 Gm. portion of a saturated boric acid solution yielded

0.3000 Gm. of boric acid upon drying to constant woight, the
0.3 Gm. of boric acid was dissolvecl in 5.5 - 0.3 = 5.2 Gm.
water. Tho amount of boric acid solnble in 100 Gm. of water at
25C. may therefore be calculated from the simple proportion
0.3:5.2: :X:I00
in which X = the number of grams of boric acid soluble in 100
Gm. of water at the temperature at which the determination
was carried out.
Calculate the solubility of boric acid in water at 25C. from the
results of the above determination.
Questions and Problems
1. What factors influence the rate of solution of bone acid in water?
2. Why is the accurate control of temperature throl.lghotit a solubility
determination essential?
3. Consult the table on page 623 of the U.S.P. and express the results of
the above determinlttioll ill Lerms of grams of boric acid soluble in 100 cc.
of water at 25C., indicating all calculations, if !iny are necessary.
 CHAPTER XII

SPECIFIC GRAVITY AND DENSITY

The absolute density of a solid or liquid is the !nass of unit

volume of the substance. Generally, hO\vever, in u:'iing the term
density, the relative density or the dcnsity at a given temperaturc
relative to the density of distilled water, either at the same tem-
perature, represented by d, or at 'loC., represented by d~: is
understood. \Vhen in the latter case, the density is corrected
for the buoyancy of the air, the number obtained represents tlw
true specific gravity of the substance. In comlllon practice, how-
ever, and in all except very exact work, the specific gravity of a
substance is taken as the ratio of the weir/ht of the 811b8iallce to
the 1J)cight of an equal volume of dl:stilll~d water at the same
temperature.
In the official standards, the specific gravity of a substance has
been established as the ratio of the apparent wcif/ht of the sub-
stance in air at 25C. to that of an equal 'Volume of distilled water
at the 8ame temperature, or di~:, except when otherwise stated and
in the case of alcohol, which should be determincd at 15.56C.
and compared with water at the Flame temperature. The latter
is the standard temperature established by the U. S. Revcllue
Department. for t.he measurement of the sperinc gravit.y of alcohol.
A considerable number of methods have been developed by
which the specific gravities of various substances can be deter-
mined. Only those methods of practical importance most
commonly used in pharrnaceutic',al anHJysis, to establish the
identity and purity of medicinal substances, will be considered
in the following pag(~s. These methods may be classified as
follows:
A. Methods used to determine the specific gravity of liquids.
1. By the use of a pycnometer.
2. By the use of the Westphal balance.
207
208 qUANTITATIVE PHARMACEUTICAL CHEMIS'l'RY

3. By weighing a solid of known specific gravity and ,veight

in the liquid.
4. By the use of hydrometers.
B. Methods used to determine the specific gravity of solids,
such as:
1. Solids hel1vier than and iJlRolllble in water.
2. Solich; lighter than and insoluble in water.
3. Solids heavier than and soluble in water.
4. Solids lighter than and soluble in water.

METHODS USED TO DETERMINE THE SPECIFIC GRAVITY OF

LIQUIDS
1. By the Use of a Pycnometer.--The specific gravity of
liquids can be determined very accurately by means of some form

FIG. 19.-Specif- FlU. 20.-Rprcngol-Ostwald

ic gravity bottle. ]Jyellomctcr (modified form).

of pycnometer or speeific gravity bottle, such as that Rhown

in Fig. 19. Although a number of different forms of pycnometers
have been developed by investigators, those described may be
used fol' practically all of the determinations of the specific
gravity of liquids required in pharmaceutieal analysis.
The SprcnyeZ-Ostwald Pycnom,eter.-Olle of t.he simplost and
most generally useful forms of pycuomet.el's is that known as
t.he Ostwald modification of the Sprengel tube (Fig. 20). This
form of pycnometer, which can be easily made by the student,
consists of a thin glass tube, 10 to 15 nnn. in diameter, to which
narrow tubes, a and b are sealed and bent as shown in the figure.
At about the middle of tube b, the bore should be slightly con-
stricted and a graduation mark scratched or etched on the glass.
Tube a should be drawn out to a point and fire polished so that
 SPECIFIC ClRAVI:!'Y AND DENSITY 20!)

the bore is constricted. Some of the commercially available

pycnometers of this type (Fig. 21) arc provided with ground-on
glass caps which fit over the eIHls of the tubes A and B. In all
except very acc:nmte work, howon'r, these caps are Ullnecessary.
The volume of the pycnometer should be about 5 to 15 ce.,
thus permitting of an accuracy of about one unit in the fourth
decimal place. When only a wry small volume of liquid is
available for a determination, howewT, a tube of smull bore bent
in the shape indicated in Fig. 20, and of a capacity of 0.5 ce.,
may be used in approximate specific gravity determinations.
In carrying out a determination with this type of.pyenometer,
it should be cleaned hy washing slleces~ively with water, aleohol,
A B
Gii!~~ )IF~-~~,.:l(,~
1

FIG. 21.-Sprengel-Ostvmlrl pyellorneter with groulH1-oll glnss caps t11.d groulld-

011 glass side tube for ust) ill filling.

and ether and then dried by drawing a current of warm air

through the 1.1111('. The pycnometer, e1eluwd and dried, is first
weighed empty. To do this, it is sut4pended from the end of a
balance beam by means of a double hook made from either plati-
num or copper wire, preft'rably the former. The pycnometer
is then filled with distilled water, whieh has been previously
boiled to expel dissolved carbon dioxide and allowed to cool,
by applying gentle suction through a rubber tube attached to
the tube b while the tip of the tube a dips into the water. The
introduction of ail' bubbles into the pycnometer must be carefully
avoided. The filled pyenOlnetcr is then suspended in a beaker
of water, along with a thennometel', [1nd the temperature is
kept constant at the desired level.
When the pycnometer and its eontents have acquired the same
temperature as the bath, usually ill about 20 min., the amount of
water must be so adjusted that it exactly fills the pycnometer
from the point of tlH~ tube a to the graduation mark on b. If
there is too much water in the pycnomete", ~ piece of filter
 210 QUAN1'ITil1'Irl:.' PHARMACEUTICAL CHEMISTRY

paprr is carefully touchpci to the tip of the tube a, thereby

drawing water from the pycnometer until the meniscus cor-
n'.~pUl1df; to tlJf~ mllrk on b. If the pycnometer is not completely
filkd. a rod or tullP carrying a drop of water is touched against
tlH' end uf the tube (I, and water if> drmvn into the tube by
eapillarity.
Thc pycnometer is removed from the bath and the outside is
dried with a doth, using cure to prevent the expulsion of water
from tho tube by expansion due to the heat of the hands, and
weighed.
Aftpr tho pycnometer filled with water has been weighed, it
is emptied, dricd onee more, and filled with the liquid the specific
gravity of which is to be determined. It is placed
in the ()(Hh;t:mt temperatme bath, the volume of
liquid it; acijust,t,d to the mark, the pycnometer is
dl'ipd with a doth in the same manner as before and
weighf'(l.
The Cieissler pycnometer or specific gravity bottle
(Fig. 22) is a. type of pycnometer well suited to
accurate determinations and can be procured in
sizes varying from 25 to 100 cc. capacity. Pycnom-
eters of this type are provided with a thermom-
eter stopper and a capillary overflow tube fitted
with a ground glass eap to prevent evaporation.
The cap should have a very small perforation in its
top to permit the liquid to expand without forcing
FlO. 22.- the cap loose. The thermometers provided with
G e i 8 S 1 e r these pyenometers are not always accurate and
PYCllometer.
should be tested by comparison with one of known
accuracy whenever exact determinations are made.
In ascertaining the specific gravity of a liquid with this
pycnometer, the thermometer and cap are removed, the previ~
ousIy cIpuned bottle is filled with the liquid which has been
cooled to from 1 to 3C. below the temperature at which the
specific gravity is to be determined. The thermometer is then
fitted to the bottle, using due precautions so that 110 air bubbles
remain in the bottle. The pycnometer with its contents is
warmed to the required temperature, any excess liquid that
exudeR from the capillary tube is carefully wiped off, the cap
is fitted to t~e side tube and the whole is weighed.
 SPECIFIC GRA VITY liND DENSITY 211

The official procedure for the determination of the specific

gravity of a fat or oil by the pycnometer method and 'which may
be applied to most liquid substances is as follows;
"The specific gravity of a fat or oil shall be determined at
25C., except when the substance is a solid at that temperature.
In this case the specific gravity shall be determined at the
temperature directed in the respective monograph and referred
to water at 25C.
"Clean a pycnometer (use a SI}l'engel pycnomet.er or a Squibb
pycnometer with a well-fitted capillary stopper) by filling it
with a saturated solution of chromium trioxide or potassium
dichromate in sulfuric acid and allowing it to stand for at least
four hours. Empty t.he pycnometer and rinse it thoroughly with
distilled water; then fill it with recently boiled distilled water
previously cooled to ahout 20C. and place in a constnnt tcmpera-
ture bath at 250. At the end of thirty minutes adjust the level
of the water to the proper point on t.he pycnometer; put the
perforated cap or stoppcr in place j remove from the h[Lth j wipe
dry with a clean cloth, free frol11lint; and after allowing to stand
for thirty minutes, weigh. Empty the pycnometer, rin:;;e several
times with alcohol and then with ethel', allow it to become
perfectly dry, remove any ether vapor, and weigh. Ascertain
the weight of the contained \vater at 25C. by subtracting the
weight of the pycnometer from its weight when full.
"Fill the clean, dry pycnometer with the oil at a temperature
below that at which the determination is to be made; place it in
a constant temperature bath at the specified temperature for
thirty minutes; adjust the level of the oil to the proper point
on the pycnometer; put the cap or stopper in place, wipe dry;
allow to stand for thirty minutes; and weigh. Subtract the
weight of the empty pycnometer from its weight when filled
with oil, and divide the differenee by the weight of water con-
tained at 250. The quotient is the specific gravity at the tem-
perature of observation, referred to water at 250."
Exercise 69
To Determine the Alcohol Content of an Official Preparation.-
It is very difficult to effect a quantitative separation of alcohol
from the solvents ordinarily employed in the manufacture of
pharmaceutical preparations, but it is comparatively easy to
212 QU,LVTI1'.l1'Il"R ['llJR:1f.1CECTIC.,lL CHEMISTRY

prf'pan' all aqtteon" di"tillnte ('oBtaining all of the aleohol in the

original mixtul'l' and to ('stimatl' tlw aj(:uhoj in the distillate by
'iome physieal m('an,~, litlf,1l as tlte d"tNlllination of the specific
.!;favity, tIll! indf'x of rnfradioll, or the boiling puint of the solu-
;.ion. 'rIte slll'eifk gradty mdhod is uffieial, and it iK probably
till' most Hecnrntn Hml :,ati:;fnetory of tIw methods nwntionccl
aLlow. This me thud e(Jn~ists of Ht~parating the nleohol from
substancf's ot!wr than water eOlltaiTlPd in tlw pl'!'paration by
distillat.ion ur otlwl' pl'lleUdUlV'; amI determining the per cent of
aleobol in lhe IlJ'('j HlI'uti(ln froIll tlw slweific I!:l'avity of the dis-
tillate. 'TIH! folluwillg notl';; whidl dpal with the methods of
tre:1till~ "arimls ty]l!.'~ of lll'l'paratioll,'; are taken from the U.S.P.:
"General Process.-l\'I('n:'<nr<: ur'cUl';thly not. leiiH than 25 cc.
nf tIl(' liquid in whirl! thp :llr:uhul b to b(: ddenninccl, nnd note
itt' ttllllpcmture. Tmlliifpl' it to [t .'il1itnhlc diHtilling apparatus
and, if its aleohul C'fJllknt is thought to be not more than 30 ppr
('('ut., dilute it with an (!(111al ,'olullI(! of distilled water, llsing the
water to rinse the VPSKf'1 that was nspd for memmring. Distil,
and ('ullpet a YOIUIllP of distillatp of about 2 ceo l('sf-5 than the
vulullle of the original test liqnid, eool or bring to the ternperaturp
at whieh the origiJlal tf':-it liqui(l Wa" mea:>ul'ed, add sufficient
distilled wat.er to melUmre exactly the original VO!tllllP of thp test
liquid and detennitHl tilt! KI}Pcific gravity, From this result
ascertain the lwn:entagn, by volume, of alcohol cont,:,linPfl therEin.
The proportion of ale'ollol by volume in the distillate equals
that in the liquid examined.
"If the liquid lllltkl' examination contains more than 30 Pl'l'
eent of aleohoJ, pro(,petl as dirnc:ttlrl nhovo, except: dilute tlw
sample with about twice itt; volume of distilled watp!, and eolleet
a volume of (E:.;tillnte uf about 2 co. 10:-38 than twice the volume
of the original tr:st liquid, cool or bring to the temperature at
which the original liquid wa:; measured, add sufficient distilled
water to measure exaetly twice the original volume of the test
liquid Ilnd determine the speeific gravity. The proportion
of alcohol by volume in this distillate, as aRcertnined from its
::;peeifie gravity, equals one~half that in the liquid examined .
. "The distillate must be clear or only Hlightly e1oudy, must not
eontain any non~vulatile material and not more than traces of
volatile substanees other than alcohol and water.
 Si'Ef'JPJ(' em..! FI1T AND f)E'I";SI1T 21~

"Thi;,; general method without modification b .,uitltble fiJI'

examining lllOf't fluid f'xtmetA alld t incturei\ provided the
capacity of the distilling flask is suffieicll t (eomll1only two
to four times tllll yolume of the liquid to be distilled) und tlw rate
of distillation is sud! that ekar di,;t.jJJutes (He proclueml. Borne
alcoholic preparation;.: will, howen'I', n'<[uirc :-:ppt'ial treat Ill(,llt
or HlP obsel'\"ane\' of special preeaut.ions to yield :-:uitahh' di;;;-
tillates. All distillates mw.;t. hI' dear or Iwarly so.
"Frothing.--Liquids whieh froth to [l. troltblpsome extent during
distillation may he di:otilled by strongly acidulating thelll with
phosphoric or sulfuric aeid, or hy the arhlition of a slight ('Xel'SS
of caieiulll chloride solution or hy the addition of a littl(~ paraffin
or yellow wax to thl' di::;tiIling flask.
"Bumping.-Liquid:'l that tend to bllmp badly WI1<'l1 IH'lti,!'d
(particularly resillous sollltions) may be distilled by making; them
alkaline with magnesia magma or by placing pieces of pmnicc
or angular pieees of metal in the distilling fh""k with the liquid,
01' by similar uwans of distributing tht~ h\~at.
"Glycerin.-Liqllids that coutain glyceriu IlIU:'lt be dilutf'd
with sufficieut di"tillecl wah.~r so that the re,.;idtlP, aftt~r distillation,
will contain at lca;.:t 50 per eput of water.
"Iodine.-All Solution" of Iodine mw-,;t he deeolori~wd, i.c.,
deprived of free iodine, bdore heing distilled, by treatm(~nt with
po\vdered zinc or by the addition of a solution of sodium thio"
sulfate. In the latter ease a. large excess of th(' thiosulfate must
be avoided and must be followed by a few drops of sodiurn
hydroxide solution to fix volatile sulfur eompounds.
"Volatile Substances.-Spirits, elixirs, tinctures, etc., that
contain nppreciable proportions of volatile materials other than
aleohol and water, such as volatile oils, chloroform, ether,
camphor, etc., are treated as follows: Mix the aecumtcly mca:,;-
ul'ed liquid with about an equal volume of saturated solution of
sodium chloride in a separator, add about the same volume of
purified petroleulll benzin and shake the mixture to extract the
interfering volatile ingredients. Draw off the separated low('r
layer and extract the benzin solution wi.th two suceessive IJortiolls
of a saturated solution of sodium chloride, using about one~half
as much eaeh time as was used in the first extraetion nlixture.
Combine the aqueous saline solutions and difltil the miJ{ture in
21.1 ()f.lN'J'17'.t'l'IT'E PILlRJfM'ECTICAL CHEMISTRY

the' ll~twl way, collecting U \'olUIl1C' of distillate having a simple

ratio to tlip \'oll1Dlc of the original liquid.
"A tro1l1Jlesmne cllluision lllay develop in the liquid mixture
upon l:ihakillg it with the benzin. In such case dilute a fresh
portioll of the origiunlliqnid with water and distil it as directed
ill 1,[H' gf'llcral prO((';'8. Thpll treat this distillate as directed
allow, ll"ing pmificd petroleulll benzin and sodium chloride
solution, lllld di"m the aqueous saline solution so produced, to
olitain 11 di~tillate which is free frolll volatile substances other
than aleohol nnd water.
"In pl'f'pnl'inp; ('(Illorlilln for distillntioll, water is used in place of
the f'ntumt!'cl ;'lulntiull of sorlium r:hloride directed abm'e.
"If yobtile ()ils [Lr(~ pr('sl'ut in small proportions only, and a
('limdy distillate is obtained, the benzin trC'atment not having
bpPll employed, the cli;;tillnte lllay be clarified und rendered
suitabll' for the SlwL,ifie gI'a vlty detenniuatioll by shaking it
with nl)(lut olle-fifth its vulume of purified petroleulIl benzin, or
by filtering; it through a thin layer of purified tnlc.
"Other Preparations Requiring Special Treatment.-Prepara-
tiolls ('ontaining free ammonia, as Arorllatic Spirit of Ammonia,
mUi't be rt~llder('d slightly acid with sulfuric acid before being
distillml.
"8Vil'it oj Ethyl Nitrite, Spirit of Glyceryl Trinilrate and
Aromatic Slilfuric Acid mUi't be treated with a small excess of
:sodium hydroxide l)f'fore being dif'ltilled. The usc of this expedi-
ent in px::tJllining Spirit of Ethyl Nitrite nceessitates that a suita-
ble correction be subtracted from the apparent result of the
:t1<:oI101 determination to compensate for the alcohol produced
during the dpcomposition of the ethyillitrite.
"Chloroform Liniment, Camphor and Soap Liniment, and
Liniment of 80ft Soap are treated with an excess of sulfuric acid
to effp(;t, (leeompnsition of the soap before thE'Y are extracted with
purified petl'olpum benzin as directed in the general process."
Object.--To Determine the Alcohol Content of Fluidextract of
Belladonna Lpaf.
Materials Required.-lO ec. of fluidextract of belladonna leaf.
A distilling (lpparatus.
A fla~k graduH1ed to hold 50 CC.
A pycnonl(ltl~r.
 SPECIFIC GR.i rI2'Y A.ND DENSITY 215
Procedure.-l, PI:wc 10 ceo of the prepar:ttion, men511red [It 25C., in a
distilling flask of about 200 (C. eapaeity, provided with tL high side tube con-
nected with :1 suituhle eOllllpll~prj add 50 ['c. of di~tilled water, distil into :l
50 ce. graduatpd flask, at ~\ieh it mte that about 48 !:e. of distillate will be
reecived ill ~i hr. j if volatile produets oiher than nleohol :n'e known to he
absent, bring to the original tlnnperature and dilute to 50 ce. with water of
the same temperature.

In very accurate work, it is bp:';t that the volume of the Rample

and of the distillate be measured at the same t(~mperature, i.r;.,
15.56C. When the temperaturps differ, the nllume of the
sample used and of the distillate eolledcd are not the same,
Hillce the coefficient of expansion of nleohol is much grouter than
that of watpr. In the yolume percC'ntages fm ukuhol, it b
assumed that the volumes al'(~ m(~asured at 15.56C.
2. Determine the specifie grayity of the dbtillflte at 25"C. hy mct1ll8 of [t
pycnometer, ::tud ihld the per cent of absolute alcohol hy volullle by refer-
ence to the ltlcohololtwtrie tflble in the U.S.P., pllge no!, lind multiply by
the dilution flwtor to find the per eent of alcohol in the original ]If(~paration.

"
The specific gravity of the distillate lllay be d(~h'l'minf'd by
means of either the Sprengel-Ostwald or the Geissler J"lyeuOlllPter
as previously described. The btter im;trurnent is the morc
satisfactory, since the tempcrature of the aleoholie Clolution can
be determined while the specifie grtlvity determination is being
made.
The method of recording the data aud ealculuting the results is
illustrated in the example:

Volume of sample at 25C. = 10 ce.

Volume of distillate at 25C. = 50 ec.
Dilution factor = 5Ho = 5

The specific gravity of the distillnte at 25/2.5 found = 0.9844.

By referring to the aleoholometric table, page 60+ of the
U.S.P., the percentage by volume of alcohol in the di::;tillate is
found to be 11 + 0.5 = 11.5. This percentage, is based on the
apparent specific gravity of 111cohol at 15.56/15.56. The
per cent of absolute alcohol by volume contained in the sample
tested is found by lIlultiplying the per cent found in the distillate
by the dilution factor, or 11.5 X 5 = 57.5 per cent. The
corresponding per cent' of absolute alcohol by 'weight may be
2Hi (j(',IN 'I'l1'. I 'l'!VI:: PflJJULtl.ECT1('AL ('IlBMIS'l'RY

fUIlIHl hy ('fHliilIlting the tablp on page H04 of the U.S.P.; e.g.,

::.7 aml 5S pf:r f'f'llt aleohol hy volume are found to ('olTl'Bpond to
~9.H) a11(150.17 per ('cut aleohnl by wpight, re;.;ppntiwly. There-
fore, fi7.f) pl'r ('put akohol hy volume cOlTe;;polHls to 49.68 per
PPllt by w('iglit.
The amollnt of nltohol prr:"Pll t in nlriulIR rn'l'paratiolls such as
fl\lidextmd~ am! tillttur('C' i" "ubjpd to cOJl"ideruhJe yuriation
dw' 10 tlw varying llrllportiolls of moisture and extractive COll-
taiw:,d ill the drug u,,;ed awl to the methods of extraction and

Fw. !!:I.-PllImmet, FIG, 2I.-"\Vcstphal specific grav- FIG. 25.-Riders.

ity b"bncl!,

COlI(~('lltration cmpJoy(:d in tlwir preparation. The official

standanl" tlwrdore, gt'IH~ndly giye a runge of alcohol content for
preparations of thic; type; 1J.(j., f1nidextmd of bPlladol1nu leaf is
required to (:olltain 57 to 63 per cent of alc'ohol by volume.
2. By the Use of the Westphal Balance.-The specific gravity
of liqnids may he determined quickly and 1:vith a fair degrpc of
aeCUl'ney by I1wans of the apparatus known as the lVcstphal
balance. 'l'llf~ mechanism of this device is based upon the gener-
ally knowll law of physics that a body immcrsc>d in a liquid is
huoyed up by a foree equal to the weight of the liquid displu('ed.
'rhis apparatus (F'igs, 23, 24, and 25) COlll;ists of it beam ba]anc~ed
on a knife edge A with !t sinker or plummet B, suspended from
one end and counterpoised by a fixod brass weight 0 at the other.
The distanee between the knife edge A and the plummet support
D j:,; graduated into ten divisions of equal length. The plummet,
 81'HCIFfC GH.1Tl'lT AND DE.VS[T}' 217
which is made of gla:-is [tIllI j" provitkrl with a tlU'rIllOlneter, iii
made of RIWh size that it will displace exaetly 5 Gw. of water at a
given temperature, usually 15.56C. The plummet and the
platinum wire by which it is suspended arc u:-iwtlly made of a
definite weight so that they are interehang('nble on different
balances.
Exercise 70
Object.-To Determine the Speeifie Gravity of a '{olatiIe ()il.

Materials Requlred.-A Wt'~tplltll lJ!lillIlCl'.

A 50 ce. cylindri('al gra(hmte.
50 ee. of yol::ttile oil.
Procedure.-l. Bal:mcf' tite apparatus in air.

To balance the apparatus, plaep it upon a firm, ]r~v('l tahle

and be sure that the leveling :-ierew in thp base if> directly hell()ath
the arm which supports the plummet, compare the h0ight of the
heam with that of the (ylinder whieh is to eontain the oil, lpngthcn
or shorten the standard if necessary, and mlju:-;t the level of the
beurn hy m('tllls of the leveling serew E until 1h(-' points F' and a
exactly coiueide.
2. Suspend the plUlllllwt in \vii.tN, eontaiIlt,d in the glass cylinder, place
the largest ridpl' weight on the hook tn whil'h tlw plummet is attaehcd, ami
hl1lanee the instrument again hy turning the leveling serew.

When determinations are made at thf'- exad temperature for

which the instrument is designed, no adjl1~tment should be neees-
sary at this point, hut in taking the speeifie gravity at 250.,
a slight adjustment if! required to eorrect for the expansion of the
liquid at the higher temlJPrature.
3. Dry ami dean the plummet nnd cylinder, flll the eyJimier with the oil
to the same level as that reached by tho water ill halancing the apparatuR,
lift the plummet. and plu('c the cylinder beneath the end of the beam,
replace the plummet so that it hangs free in the oil without touching the
cylinder wall, and place the riders on the beam until balance is restored.

The temperature of the oil (25 DC.) at which the final adjust-
ment of the balance is made should be the same as that of the
water used to standardize the apparatus. The specific gravity
is read directly from the position of the riders on the beam i
e.g., if the largest rider is at 9, the second at 7, the third. at 5,
218 Q[UNTITATIfE PHA.RMACEUTICA.L CHEMISTRY

Hnd the fourth at 2, the specific gravity of the oil is 0.9752.

Hf~,uliIlgs should alwfiYs be made to four decimal places; i.e.,
all four sizE'S of riders should be used.
3. By Weighing a Solid in Air, in Water, and in the Liquid.-
:PhllllllHt,S ur :o;inker;:; of nuious forms made of sealed glass tubes
partly iUled with llwreury or of S()nH~ metal such as platinum
have bpen dpvised for s!H'C'ifie gravity determinations of this
typP, hut (l hras;; wf'ight will suffice for the determination of the
,specific gra\'ity of liqui(ls wbieh do not attack the brass. For
example, if a piuee of bratis weighs 10 Gm. in air, 8.82'1 Gm.
ill water, and 9 Gill. in a Ijquid of unknowll specific gravit.y, the
f:'ll(!r'ifie gravity of the last call he calculated from a simple
proportioll as folluwi;:
1.17u:l::1':X x = 0.85

where l.l7U is the 10";'; of w('[ght ill wutt'l' (10 - 8.824 "= 1.176)
or 1he weight of tltp water disp\ltC'cd; 1 is the loss of weight in the
liquid; 1' . the specific: gravit,y of water; and X, the speCIfic grayity
of t.he unknown littllid.
The prineip!P5 involved and the method of making the deter-
mination art: similar to that gi,'cll in detail under the determina-
tion of the specific gravit.y of solidt> heavier than and insoluble
in wuter (pagf' 219).
4. :By the Use of Hydrometers.-Hyclrometers are instruments
used to determine q\lickly the n.pproximaJe specific gravity or
density of liquids by flotation. They usually consist of a sealed
glas;:; tube weighted with lllercury or shot and frequently are
provided. with t1 tlwrmometer so that tho temperature at which
the specific gravity is dptl~nnitw(llllay be read at t.he same time.
They an' variou,sly called alcuholomctCl's, lactomcters, 'urinometers,
ete" accurding to the use for which they are designed. They are
either constrncted to read specific gravity directly, as, for
example, t.he hYllromet.ers for general use and lactometers, or they
are graduated aceording to some arbitrary seale of degrees, as
in the case of the Baumc and 'fwadell hydromet.ers. Specific
gravity hydrometers are usually made in sets of four or six, each
illBtrument being calibrated to cover a different range in specific
gravities so that the complete set will suffiee to determine the
specific gravity of almost any liquid; e.g.) the different inst.ru-
 SPECIFIC GRAVITY AND DEXSITr 219
ments in the set of foul' are calibrated to rc'ad from 0.600 to 1.000.
from 1.000 to 1.400, from 1.400 to 1.800. awl from 1.800 to 2.200.
Vilhen making n, specifie gm \"i ty d!'tI'rmi1HllilHl with t1 II ydl'Olll-
eter, the thoroughly cleaned imtrullwllt, should b{_~ :dl<;\\"cd to
sink in the liqnid gl'uduuIly until it is at re:;t, HU'll dq)n';'i~l'd
about 1 in. to moisten [1 portion uf till' skm, ulluwed to eOlne to
equilibrium, and the reuding taken.

Exercise 71
Object.--To PreparE' 200 cc. of Sulfuric Aeid Containing
10 Gm. of H 2S0 1 per 100 ee.
Materials Required.~20 (ie. of eonc(,lltrated ILSO.j.
A hydrometer enlibl'ntcd from specific gmvity 1.000 to 1..100 :Lt, 25C.
Procedure.-DiIut() 20 ceo of Illll"l.\ cou('f'ntrated ~ulfurie !teid 10 IOO ("c,
with distilled wat.er, allow the resultiug; solution to at'quire a telllpera1urt
of 250., and take its specific gravity as accurately as pO~5ible with the
hydroIIJett,r.

If the specific gravity of the solutioll is found to be 1.2603,

it contains 35 PPl' cent of H 2S0 4 as found by l'ofl'l"l'ing to tlw
table, page 615 of the U,S.P. The weight of the prepared ~wid
2 X lOO = i)~7 .1'430 G 111.
. t,0 b e (iiI nteeI WI'II t Ilell 1.ue 10 X 3~
so1utlOll
,0
This weight of solution containing 35 per eent of H 2S0 4 when
diluted to 200 ee. will fol'IU a solution containing 10 Gm. of
H 2S0 4 per 100 ce.
Solutions prepared as directed above are subject to slight.
inaccuracies of concentration, since speeifie gravity determina-
tions by means of hydrometers are not exact, hut if the specific
gravity is determined accurately by means of a Jlyenomctcl',
the eonecntratiol1 of a solution prepared in this manner will be
sufficiently aecurate for alI ordinary qualitative purpose,.;.

METHODS USED TO DETERMINE THE SPECIFIC GRAVITY OF

SOLIDS
1. Solids Heavier than and Insoluble in Water.-A. Weigh
the substance in air and then in water.
According to the law of Arehimedes, a body sw:!pended in a
fluid is buoyed up by a force equal to the weight of the fluid
displaced. The weight of a solid in air, therefore, less its weight
22(1 (JCAN1'1'l'.!TlI/~' ]'ILllDf.{('IIr.'l'J('.IL ('llRJ1IS1'RY

in watl'l' gin':; 111(' \wight uf an eqnal \"Illume ()f walt'!", since

trw w,ll1n~H of watN di;:plrU'l,d i;-; eqnal to tllP volt1l11r of the
,':fllil]. If tIll' WIight in air j,,) djyjrhl by the In.~.., of mjght in
\I":ll('l', th(~ 1'1':<11lt i~ tIll) spl'eific grayity of tho f'llbstallC'(, nt the
tC'lllIwmt1ll'P at which thl deh::l'll;inatioll is made. For example:
SllPPO~P t hat a pi(~ee of IHt!":-i IH'igh;.: lO.OOlO Om. in air at 2f5C.
amI 8.8010 GIll. illunen.;ed in wab.'r at 25C. The weight ill air
di l'idl'll hy the los:,; of w('ight, 1.2030 GIll., gin's the specific
~mvity of tbe 1ml"s at 25/25 or 10.0040/1.2030 = 8.:32.
B. If thc' :mh:-;tallC(; is in slUall picces ur i" [l powr\('l'. thp specific
~ra vity may lip determillc'd a" illllstrntpu ill the folluwing exercise:

Exercise 72
Object.~T(J DeU;l'llline tIl<' SIW(jfic Gml'ity of Powdered
Bal'illlll fhdfatl"
Materials Required.-i to 10 UIIl. uf jlowdered bariulll Rlllflite.
A :Hn:dl YUilIlU(;i:l'il! Ha~k (25 or 50 (~e.l.
Procedure.-l, 'iVl'igh abullt 10 Om. of the powderr',1 bariulll Hulfatfl
al'l'luatdy,
2. A(:('lImt,dy wl'igh the .'';llwll \'olmfll'll'ic fl!\~k filled pxndly to t.he>
gI'luiulltioll lIlllrk wit It l'l'nentl_,, hoiled ,.li~tillwi wllter at 25C.
:.I. Illtr,,'IIH'{~ the powr\{;!'(',l harium Hulfat" into the flask qllan;.itlltivl'iy,
rl'1I1m'" :Illy air ol'ellldt'd ill tlte powder by lll!;itlltillg (he mixture, draw off the
waH;l' wit It a pipette until the 1,,\'1'1 of the liquirl is again exaetly at the
graduatioll lIlurk Wl!l'll tlll' liquid is at 25"C., aml weigh the fllLsk and its
"OUt.Pllt, a!"lIrall'!Y.

It is e\'idput. that. II \'lllllme of wuter eqllul to that of the harium

~tllfnt[! ha" bE'f'1l displaced and removed from the flask. The
~Iweifie gravity of the harium :mlfate Illay then be calculated, f'.y.:

Wr,igilt of powdered barium sulfate = 8.0 Om.

\Vpight of fla~k fillt'd with wateI' = 80 Gm.
W('ight of fia"k, barilllll sulfatp, amI water = 85 Om.
80 + 8 - 85 = 3 Gm., the w(ight of the volume of water
displaced by trw hariu,Ill Hulfttte; and % = 2.6666, the specific
gravity of the barimn sulfate.
2. Solids Lighter than and Insoluble in Water.-A. By Weiyh-
ina in rVater.~If It suhstance is lighter than water, it will float,
and its volume cannot be obtailwd by weighing it in water unless
it is completely submerged by nWfLUS of a weight or sinker heavier
 SPECIFIC' GRi1 nIT AND DEXSI1T 221

than WittN. The method of determining awl calculating the

specific gravity of it :mh::;tancc of this type is illustrated ill the
following exercise:

Exercise 73

Object.-To Determine the Specific Gravity of Camphor.

Materials Required.-A llieee of ClUllphol' (2 to 4 G1I1.).

A suitable ~inkel' such aH a 2 GIll. weight.
Procedure.-1. 'Veigh the piece of e:tmphol'in nil'.
2. 'Veigh tlll' siuker immersed in wItter.
3. Att:wh tllC' sinker to the piece of cllInphur IUlll Jiml lIH~ir comhinetl
weight when illlllwffled in watcr.

Since the camphor iR lightf~r than water, t.he weight of the "illlwr
an~l camphor is less than the wpight of the sinker alone. Tho
differenee between tIw weight of the Rink!'!' ulld camphor in water
and the ,veight of the sinkC'r alone in wllter plus the weight of the
camphor ill ail' is a llWttSure of the watp!, displaeed by tIll' CampllOl'
and also a measure of it8 buoyant power, e.g.:

"Weight of the camphor in air = 4 Gill.

Weight of the sinhI' in Wllt<~r = 1.6 Om.
Weight of the eamphor and sinker in water = 1.5 Gm.

The difference between the weight of the ;-linker in water and

the weight of die Hinkel' uuel camphor ill water = 1.6 - 1.5
= 0.1 Gm.
The weight of the camphor in air pIns thi" diftprenee of 0.1 Gm.
is the weight of a volume of water equal t.o the 'weight of the
piece of camphor or 4 +
0.1 = 4.1 Om. water displaced.
Since the spccific gravity of a substance is found by dividing
its weight by the weight of an equal volume of water, the sp('eific
gravity of the eunphor is 4/4.1 = 0.97.56.
B. By the Flotation ~il-fethod.-Thi:-: llwthocl eOl1sist:-; of bringing
t.he substanee into equilibrium with a liq lliel of krlO1;vll spreifie
gravity. It is used in the offieial detel'lnination of the specific
gravity of suell substances as white and yellow wax and sperm-
aceti and by food and drug analysts to determine the spedfic
gravities of certain fats and oils.
222 QUANTITATIVE PHA.RM~1CBU'1'ICAL CHEMISTRY

Exercise 74

Object.-To Determine the Specific Gravity of Yellow Wax.

Materials Required.-Abollt 0.25 Gm. of yellow w:.x.
25 ce. of 11leohol.
A pycnollwter.
Procedure.-l. "Melt the \Vax at u low temperature and allow it to fall
into separated drops from jl'1!t above the sUffaee into al('ohol that has heen
warmed to from 45 to IlO"O. Allow the globule:; to remain in the alcohol
until it IU1S cooled ~Jlontan('ously to roolll temperature (20' to 25"0.), then
remove the \VllX and keep it ut room temperuture for twenty-four haUl's."

The meltf'd wax separates in glohules free from occluded air

which solidify whell the mixtme is cooled. The v.-ax which has a
high eoefficient of expansion is kept at room temperature for
24 hr. to permit the globules to acquire their normal volume.
2. "Prepare a mixture of fOllr volumes of nleohol lind enough distiJ.lecl
water to make ten YoluInes, and allow it to stand until free from air bubbles.
Moisten the globules of WILX with clisWlcd water by means of a brush, and
place them by mClU1S of forceps in the aleohol solution jURt prepared and
cOl1t,ained ill a lwaker. Then add alcohol or air-free distilled water, as
required, to the mixture kept at 25C., until the glohulcs of the Wax flout
and rest indifferently in the liquid. Finally determine the specific gravity
of the alcohol-water mixture. The figure thus obtained represents the
specific gravity of the Wax eXllmilled."

The wax globules are moistened with water :-;0 that the water-
alrohol mixture will COllle in contact with all portions of the
globules without occluding air at the intersurface. When the
globules are in equilibrium in the alcoholic solution, their weight
must be equal to the weight of liquid which they displace or they
would rise to the top of the liquid and float or sink to the bottom
of the beaker. Consequently, when such equilibrium is attained,
the specific gravity of the wax and that of the liquid are equal,
and a determination of tlw specific gravity of the liquid is a
measure of the specific gravity of the wax.
3. Solids Heavier than and Soluble in Water.-The specific
gravity of solids heavier than and soluble in water may be deter-
mined by weighing the solid in air and then in some liquid of
known specific gravity in whieh the solid substance is insoluble
such as oil of turpentine, chloroform, benzene, etc. Thus, the
weight of a piece of alum in air divided by its loss in weight when
 SPECIFIC GRl1'VITY AND DENSITY 223
weighed immersed in oil of turpentine gives the density of the
alum relative to the oil of turpentine, and if the specific gravity
of the oil of turpentine is known, that of the alum can be cal-
culated, e.g.:
Weight of the alum in ail' = 10 Gm.
Weight of the alum in oil of turpentine = 6.5 Gm.
Specific gravity of the oil of turpentine = 0.860
10 - 6.5 = 3.5 GIll., (oss in weight of the alum in the oil of
turpentine equal to the weight of an equal volume of the latter.
:3.5:X::0.8G:l x = 4.070 Gm.,
where X is the weight of a volume of water equal to thc volume
of the oil of turpe!ltill(~ displaeed, and 10/4.070 = 2.4570, the
specific gravity of the alum.
4. Solids Lighter than and Soluble in Water.-The specific
gravity of substances lighter than and soluble in water may be
determined by weighing the substance with a sinkPI' in some
liquid of known spccifie gravity in which the substance is not
soluble. For example, the specific gravity of lithium may be
determined from the data below as follows:
Weight of the lithium in air = 2.0 Gm.
Weight of the sinker in air = .5.0 Gm.
Specific gravity of the sinker (brass) == 8.4
Specific gravity of liquid (kerosene) = 0.830
Weight of lithium and sinker in kerosene = 3.8 Gm.
The density X of the sinker referred to kerosene as a standard is
0.830:1 ::8.4:X x = 10.1205
The weight Y of the sinker in kerosene may be calculated,

-~.~.-. = 10 1205
..5.0 Y = 4.5095,
- Y .
and the density of lithium compared to that of kerosene is
2
2 + 4.5060 - 3.8 = 0.7391.
221 ("t'.lN'l'I1'.l'J'IFE l'H.lRJf.H.'EU'l'/CAL C'llK~[J8'l'RY

The sppcifie gravity Z (If thE' lithium iB, therefore,

o.s:m:I::Z:O.nOI
Z =: (Hil35 , the ~peeiiir gmvity of tlw lithium.
Questions and Problems
1. Define till' t('rIllS ~IWl'ifi(' gravity, a]lpul'Pllt ;;pncitic gl'l1vity, true
I'pPeiti(\ gradlY.
2. By wh:l!. rliffen'llt lllnthofls might the ~pecifil: gwvity of ulcohol be
det!'rlllilH'd?
3. A I () Gill. weight. of hntss, HIll:!'ifj('. gravity 8.'1, SUIJlll('rged in glycerin
,,"I'ight'd 8.51:35 Gill. What i8 tho ~J.leeilie gniVity of thn glycerin?
4. Gin'll tho s[w('iti(~ gravity Df sulfuric Heid at l.5 C. ilS 1.8,122 l'OIupllred
Q

to \vUI,'!' Itt 15 c C., what i~ it;; ;-;pef'ific gravity referred to water at 21 e.?
(Brie U.S.1'., ]lag" fill.)
5. How hlni'll nit ric lldd, spp('ilil~ gravity 1.45, nnd hOlv lllllCh wah~r Illllst
Ill' lllixe(ltn furm 1 l. of nitric:: add, ~jJl:eilie gravity 1.20?
6. Calelll:tte the ~peeitie gmvity of a piece of sHlldalwoud frolll the fullow-
ing data:

Wpight ill' air =' 1O.10() Gill.

W!'ight of Hinhr in waiN = 12.200 Gm.
Wt'ight of ~illkcr awl s~1l1(h!l\l'l)ot! ill water = 1UlOO Gm.

7. How wany gr~lI!)l' of Ivater will a llyellOTnctlr tmlibrated to hold 100

Gm. of water at. 25"0./25"0. tiUfI 7(\0 mm. Pl'PSl;Ul'P contain at. 190. und 740
lfllll. PI"e:;~ure? (See U.8.P., ]Jngf~ 022.:'
8. Give the 8pedfic gravity of l1kohol eompm-ed to wtlter fit 15.560. and
700 1I11l1. prp~~nre ill a Holution contltilling Hi.5 per Cf'ut of alcohol by volume.
(Sef~ U.S.P., p:lge GOt}
9. Briefly state how ttll' ~Jlceitie gravity of cadi of the fulluwing might he
ddermined with cOlllparative ae(!UJ'[tcy: olive oil, benzene, powderer! hor11.x,
gl'lLllular zinc, aue{ C!WllO butter.

The approximate Rpecific gravities for many official substances,

Bm~h as acetone, acids, and llumerOUH preparations, are given in
the U.S. Pharmaeopoeia and National Formulary simply as
information for the conversion of volume to weight. 'When the
minimum or lIln,ximull1 speeific gmvity or a range of ~pecific
gravity is given, it is a definite constant of value in
df'terll1ining the identity, purity, und concentration of the
BubHtal1ee. Those Hubstallecs for whirh definite specific gravity
limits are given in the official stundardi'! are Bhown in the following
table.
 8l'EC1FJ(: (ilL! nTr .t.\']) ])ESSl1T 225

~pr:~ifi~' [ ;"IrJl'I~lfl~'

-_.
Sub;-;T,n.ll(!l' p;rdviry. t'ulJ,:,t all'_'(' gl_':',.Yity~ (

U.S.P. 2,,'(:. U.s:;~----------- 'I~~'~

A.chl, n.cetir~, J!-ladal. .. T." 1.Oi:ito 1(1)50 d".r~'l.ti 1'011' J11.'(_llt,~ " , . '1(1.8,~ja1.n I) ,Sri')
Alenh"l (at J5.iili'r'.) ...... (n.m.t.) !J.SHi "u,ahp tus . . . . " . .,. Ii. (1(1[. to II. ~l~~,
Alc~}!':'al~. dphydratf!d I"HOP1 ., . , . . . . . _[O.!tft:l1U n_!!7~j
(1;",.6 C.) .............. (run.t.) 0.7:18i julllI"'t'. .. .. , ........ O.8i,4 tn 1).8;(1
Alrolwl. ,1i\utrd (lr,.~(;c.)
Amyl nitritT~ ....... _... , ..
I). !J:l" to o. n:l7l'
0.805 to o. 87.51
Illv""d,'r .............. '1.
~iJ to O. 888
If'Tn.u~.: ..... , , , ....... 0, 8.4~. 1 t'.,1 n 8ilfJ
Bal.urn. l'rruvirUl ......... 1. 15() In 1.170 mllst"rol ...... , ........ 1 .01.J to I .02fl
'Ben7.in, petroleum, puriftpd (). G3,1 to (). GnO myri~t.i.ti.l.. .. , ........... O. S.i~! tu (J, ~r.!4.
Brandy .................. O. (Kja to 0.021 umngf! .............. , .. O.84~ tu O.Mn
Bromine, R.I .............
Carnuhof, ~pirit of, .... ~.
, (n.1.I.) ~1, OUt)
O. 8.24 ~.() 0, H2t)
'1
eP.penllint .. , , ....... 0 , 8!.lt.') to. (). ~. j.)S
rllse 4.... , ~ ............ O. 84S tiJ (\ . Ht):1
p.

Carbon tr.tmchloride ..... ' ]. il88 t(l 1. Iinn ro~!'m;lfY .............. O. S~I.! to \), n12
Chlot{)form ....... ' " .... 1.474 to 1.478 santa\. .............. II. !ttl'-, t.o (I llsn
Collodion ................ n. 705 to n. 77:::1 I.ijl""afJ;ts ....... , .... ,[l.Uij['1 to 1.077
COP[~ibll ...... , .......... Il.U30 to (l.~t~H;1 "'pp.(lrrnAIQ ....... , l),U17 tn U WHo
Creosotc> .... , .... , .......
(:ref.lf~ote (;arbtJIllltp.., ......
(n.l.t.) L(}7tJ
Cn.l.t.)] .14.1),]
I 1-11', rl'(~hfi('ti., . . . .
tUfJWutlllf. ..
'10\1(,0 tit n 1mo
n 854 to II aDS
Cr('(',,\ ................... 1. (I~O t" 1 . OM'I lurp,ntm,. n\,tlfi~d. 0.8;;:1 til II !\I',~
Ether ...... ' ........... n.71:1 100.711:1 Olr"If'swJ!fllepidium (nl!.) 1.0
Ether. "b,,,I,,t,,. R....... (n.m.!.) 11.71011 Ox hM .......... ' 1 O!,-, tn 1.1)~;;
Ethyl "I.. tnl<'. I~ :. .... () 8!1:J.tu (I.,fi;~FI\' Par,dlin, "hlorinald. 1.0n tu 1.07
Ethyl mInt!', .pmt of .... (n.Ill.,.) n 8:..1\ l'mr"lemn'... .... 11.820 to (L 8(\5
~thyl oxi,le .... .... .., n. 71:l!" O. 711 l'<rtr,,}utum. li'lUid ........ l). 8~5 tn (). un;,
hut"lyptol......... .... () !l21 In fl. H2:1 HOHi" ........... , "" .' [. (17 to 1 m
Eng"",,1 ... ... '" " 1.llbli tn 1 .07f'i Sp!'flt"'tI'eti ............... I). (1:18 to II. f)a
}"f...rrie l'11!oridC', r=,f)]ution vf ]:!U to t.:.;2 I SwHum hYl)f){'}J]f)]'itl', dj~
Glyc<rrin ................. (n I.t) 1 :H~): luted solutill" "f. ....... \1I.1.t.) 1.20,;
Glyceryl tl'ini!r"t". "pirit <If (\.814 lo o. 8~OI Tar. i I111iIlI'l' .............. (I. \l50 tu 1.05"
Honey'. . .. ............. (n.Lt.) 1. fl!l(I T"rcher", ................ O. 81:\11 to 0.81'1.;
lIllllt, I'''",et nf .......... 1.350to 1.4:101 Wax. wl,ile and ydlnw ... O.8r,lJtoO.!lr.O
Methyl ".aliCIHte ellat urn] I 1.17(., II.) 1.182 Whisky .................. o. U:;o to 0.923
Methyl Rlllicylatn (synt!II'!- N.F.
ic) .................... 1. 18U t.o 1.18,\ And.hol.. ................ O. !18.1 to O. (lS7
Oil (fix,,,) of . I Ether, "pirit of .......... ' 0.784 to O. 7()4
almond ."prJ'socli. " .... o.. ()W to O. \lU,l E. thYlll""'t,."t'" ............ \Il. 8.92 to 0.898
Castnr .. , . , .......... O. U45 to {I. fI(iGlr\ Cal'lun.d., .... ,., ...... ~. (H.1.t.) 1,:-{5
chaulmoogra ........... 0.940 to O. HfiO!1 H:trn~~mdi~ water .. ' ...... 0. U7n to 0.082
<o,lliv"r, .............. IUn8 to O. fl27:1 Oil (fixed) of
corn, ................. , tLU14toO.n21~ ('I"Ot01L...'IO,H!l;)tl)O.U.)O
r~ott.on!'!r.r.d ............ , O.Hpi tt.,O.021;. !'f'N:l1Y.lf'! . . . , . . . . O.111l\ toO.U21
linsE'l~d., ............... O.02fJ to O. U8f11 Oil (yolatilt~) of
O.
olive ................. !IIU t() n. ()! ;"1 bil"'}< (ar. fl'"tifie,l ...... 'jl). 881i tn O. \);;0
thoIJbl'nma' ...... .... O.HG8 to 0.8()41 hiUe!' nrclllf';" ........... 11.815 tn 0.8,,1
Oil (volatilel of I "awway .... , .......... O. (101) to 0.1110
anise .................. 0.1l78 to O. \)1'18.1 ecmlulIlom ............. I). m 7 to (I. !l47
bitter uln!ond .......... 1. O:lR to I. ~I).
chenopodlUm ........... (n.l.t.) 0.9.)0
0Il lllYf'.'ill ....... , .... , .. IJ. 962 to 0 !!llD
orang'> fi()WerR .......... n.863 I." D.8SI)
cinnamnn ............... 1.040 to 1.063 !limen!" .... , .......... L{ll8to 104.8
e!ov\, .............. , ... II.
ms to I.OO()I'I thymI' ................. 0.894 to O. !)30
coriancler ............ '10'
&j~l to (I. 875 T('(m,hlnrel.h~I"tll1.. I. non t.[J 1. 61tl

(n. m. t.) = Il'Jt more than. (n.]. t. ) = lint 11'.,. th"n. It. = reagent,
1 Saturate-Ii with watf;r.
2 Diluter! with 2 tim(~~ its volume of distill(~d water~
3 At 100C. compared to wntr.r Ht 25C .
At 30C. complred with water at 15C.
At. 50C.
 CHAPTER XIII

MELTING, CONGEALING, AND BOILING POINTS

Melting Point.-Whcn an amorphous substance is heated, it
gradually softens and eventnally acql1ires the properties charac-
teristic of a liquid, but no definite point of transition from the
solid to the liquid state occurs during the process of heating.
When a ery~tallille solid is heated, there is a sharp change from
tlm solid to the liquid state at a definite temperature, this temper-
ature being known as the melting poz:nt of the substance.
The nH'lting point of a compound may be defined a8 the tem1Jer-
ature at which the liquid and solid phases of the compound are in
equilibrium. Each pure fusible compound
has a fixed and definite melting point, and
this constant is often used to determine
the purity of a solid, since the introduction
of even a small amount of impurity lowers
the melting point to a considerable extent.
If appreciable quantities of impurities are
present, the melting point is not sharp but
ranges over a number of degrees and is
usually 10,y. In the official standards, the
melting point of a substance is defined as
the interval of temperature within which
the substance is observed to melt when
treated according to a definite method.
:r. . lG. 26.-Appamtus for 'L'
taking melting points. .[' or the determination of the melting point,
the official substances are divided into two
classes, namely: (1) substances which can be readily reduced to
a powder; (2) substances such as fats, fatty acids, waxes, and
paraffin which cannot be reduced to a powder readily.
The apparatus (Fig, 26) required to determine the melting
point of either class of substances consists of the following parts:
(I) a thin-walled test tube about 3 or 4 em. in diameter and about
226
 MELTING, CONGEALING, AND BOlLING POlN'l'" 227

10 em. long; (2) a iitirring rod b(!]lt into a eirde at one end to fit
the above tube and at all angle at the other Plld to permit easy
manipulation; (3) a standard thermometer eovE'ring tlH~ desired
range of temperature; (4) an auxiliary thermometer to take the
emergent stem eorrection, prefrrably graduated from 20 to
100 0 e.; (5) a capillary glass tube about 6 em. long and 1.0 mIll.
in diameter sealed at one end.
For temperatures up to 200C., a purified, concentrated iiulfnrie
acid is a suitable hath. For higher temperatures, up to about
3500., a pure grade of cottonseed oil (almo::;t colorless) will
serve for a limited l1umJX'r of drtenniImtioIls. Other, though
less desirable, substitutes for sulfuric aeid fDr use at high temper-
atures are: (1) a pure grade of pamffin which has lwcn fre::;hly
distilled; (2) dean, white, hydrogenated cottonseed oil. A very
satisfactory hath is prepared by cautiously boiling together, for
from five to ten minutes nnder a hood, a mixture of 70 parts of
sulfuric acid and 30 parts of potassium sulfate, stirring constantly
until the pota.'1fliulll flulfatc is cornpletdy dissolved.
The following exereise illustrates the me thuds used to deter-
mine the melting points of official substances:

Exercise 75
Object.-To Determine the Melting Point of Salicylic Acid.
Materials Required.-A llH'lting point nppamtus as deseribecl above.
20 to 30 ce. of eoneentra tE~d sulfuric acid.
About 0.05 Grn. of salieylic: acid.
Procedure.-l. Reduce the salicylic acid to 1\ fine powder and dry it at
100C. for 2 hr.

The material is dried because substances which contain water

of crystallization or moisture must be renderNI anhydrous to
prevent liquefaction when heated through the solvent actioIl, of
water.
2. Place sufficient sulfuric acid in the test tube to fIll it to a depth of
about 5 em. so that the upper portion of the t.hermometer bulb Ulay b('
immersed about 2 cm. below the upper surfacc of the bath and leave about
2 em. of the liquid below the lower end of the bulb.

This arrangement prevents unequal heating of the different.

parts of the thermometer bulb.
228 QUANTI'l'ATIYE PHARMACEUTICAL CHEMISTRY

3. Introduce enough of t.he lin ely powdered dry salicylic acid into t.he
(:tpillttry tl\btl to form a columll about 0.3 om. in length.
Tht: salicylic acid may be packed into the scaled end of the
tube by tapping the open end of it iuto a little mound of the pow-
der, then inverting the tube and tapping it gently on a solid
surfaee, or by dmwing the broad side of a triangular file across
the surface of the tube just below the I)owder.
4. Attach the charged capillary tube to the thermometer by wetting
both with the acid of the bath, and the tube will then be held in position by
ClLpillary attractioIl, or attach the capillary tube by IllenllS of it rubber band,
dipped from a piece of rubber tl\bing, in such a position that the column of
balieylic acid is centmlly located by the side of 1,he thermometer bulb.
Attrrch the lLllx:iliary thermometer so that the centcr of its bulb is a~ close as
possible to the stem of the main thermometer at a point midway between
the HurftLee of the b:Lth and the 160 graduation lllark. Heat the acid bath
by mcnns of a frec 13IlIlSCll flame until it temperature of about 140C. is
l'el1chcd, then carefully reguhLte the rise in temperatul'e to about 3 PCl'
minute until the substance begins to melt, and then regulate thc rise ill
ternp('l'ILtUl'C to about 0.5 per minute until tho snlieylic acid is completely
melted, while stirring the hath continually. Record the melting interval
tempcmture aud the temperature l'egi~tcrod on the auxiliary therlllometer
at the end of the mclting.
'Ehe temperature at which the column of salicylic acid is
observed to collapse definitely in the capillary tube is considered
as the beginning of the melting, the temperature at which the
salicylic acid becomes liquid throughout is considered as thc
end of the melting, and the interval betweon the temperatures
at which the melting begins and ends is taken as the temperature
rangc of the melting point of salicylic acid.
The rate of rise in the temperature 01 the bath can be regulated
easily if the bUl'l1er is held in the hand so that more or less heat
can be applied as required. Since heat from the burner may
affect the temperature registered on the auxiliary thermometer,
it is advisable to construct a nlOvable platform of stiff paper
around the main thermometer and adjust it to a position just
below the bulb of the auxiliary thermometer.
'1'he temperature observed on the auxiliary thermometer is
used to correct for the contraetion of the thread of mercury above
the bath by the following formula:
Correction = 0.00015 X NeT - t) where N represents the
number of degrees from the surface of the bath to the melting
 MELTING, CONGEALING, AND BOILING POINTS 229

point, T the temperature at the end of melting, and t the temper-

ature registered on the auxiliary thermOIlleter. The correction is
added to the observed reading of the main thermometer; i.e.,
if in the determination of the melting point of salicylic acid, the
main thermometer is immersed in the bath to the 30 0 graduation
mark, the acid is completely melted at 156.5C. and the tempera-
ture registered on the auxiliary thermometer if) 40C., the correc-
tion for the melting point temperature would be
0.00015 X 126.5(156.5 - 40) = 2.59C.
and the corrected melting point becomes 156.5 + 2.59 = 159.1C.
The melting I)oint of a substance which cannot be readily
reduced to a powdcr is determined in the same manner with the
following exceptions: The substance is melted at as Iowa temper-
ature as possible and drawn into a capillary tube, open at
both ends, to a depth of about 1 cm. The charged tube IS allowed
to cool at 10 0 e. or at a lower temperature for 24 hr. or in contact
with ice for 2 hr. The melting point of the substance is then
determined by attaching the charged tube to a standard ther-
mometer as described in the above exercise or by means of a
rubber band and heating in a suitable bath so that the rate of rise
of temperature dOe.'! not exceed 0.5 0 per minute for the last 5
before the expected melting point is reached. The temperature
at which the substance is observed to rise ill the capillary tube
i~ taken as the melting point. Melting point determinations
by the above method arc generally peliormed on substances
which are mixtures and whieh do not give the sharp melting
point characteristic of pure organic compounds.

Questions and Problems

1. Look up the mel~illg points given for each of the following substances
in the U.S.P. and explain briefly why an interval of melLing point tempera-
tures is permitted in some cases while fixed melting points are given in
others: Ca) paraffin, (b) petrolatum, (c) acetylsalicylic add, (d) menthol,
(e) cocaine.
2. Why should thermometers for use in melting point determillat,ions be
made of annealed glass? (See U.S.P., page 470.)
3. How might the emergent stem correction for the main thermometer be
determined by interpolation from a correction curve when a number of
melting points are to be determined? (See U.S,P" page 457.)
230 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

T.A.DLE XXXI.-MELTING POINTS OF OFFICIAL SUBSTANCES

Melting Melting
Substance point, Substance point,
C. C.

U.S.P. U.S.P. (Cont.)

Acetunilid ................ 113 to 115 Phenobarbital. ......... " 173 to 177
Acetphenetidin ......... " 134 tD 135 Phenolphthalein.. . . . . . . . . (n.l.t.) 258
Acid, ucetyI81tlicylic..... .. (n.l.t.) 185* Phenylsalicylate...... .. .. 41 to 43
Acid, benzDic ........... " 120 tD 122 Phloroglucinol, R ......... 2Hl to 219
Acid, salicylic ............ 157 tD 150 Pilocarpine nitrate ........ 170 to 173
Aminopyrine ............. 107 to IOn Procaine hyrll'ochlorirlc .... 158 to 156
Antipyrine ............... 111 tD 113 pYl'OguUol. ............... 130 to 133
Atl'Opinc ................. 114 to 116 Quinine ethyl""rbonate.... 89 to 91
AtrDpine "ulfate. . . . . . . . . . (n.Lt.) 188 Resorcinol. ............. 109 to III
Barbit:t1 ................. 187 to HlO Sacclwrin.. .. . . . . . . . . . . . . (n.l.t.) 222
Benzidine, R ............. 127 to 120 Santonin ................. 169 to 171
Betanaphthol. .......... " 120 to 122 Scopolamine hydl'obromid" 190 to 102
Caffeine ................. 235 to 237 SpermaCf!ti. . . . . . . . . . . . . .. 42 to 50
Camphor ................ 17'. to 177 Sud, prepared .... , . . . . . .. 45 to 50
Carbromal. . . . . . . . . . . . . .. 116 to 117 Sulfone thylmothane. . . . . .. 74 to 76
Chlorbutanol.. .. . . . . . . . . . (n.l.t.) 78 Terpin hydmte ........... 115 to 117
Cocaine ................ '. 9() to \)8 Theophylline ............. 269 to 272
Cocaine hydrochloride..... (u.l.t.) 183 Thymol. . . . . . . . . . . . . . . . .. 48 to 51
Codeine ('LIlhydrou") .... " 154 to 156 " Trinitrophenol. ........... 121 to 123
Colchicine ............... 142 to 146 Vanillin.. . .. .. . . . .. . . . . .. 80 to 82
DiehlDrnnline. . . . . . . . . . . . . Cn.) 80 Wax (white and yellow). .. 62 to 65
Dinitrophcllylhydm~ine R. 198 to 200 N.F.
Diphenylnmine. R ........ 52.5 to 53.5 Arecoline hydro bromide ... 169 to 171
Ephcdrine. . .. . . . . . . . . ... 34 to 40 Camphor, monobmmated. 74 to 76
Ephedrine hydrochloride .. 216 to 220 Chlorthymol. . . . . . . . . . . .. 59 to 61
Ephedrine sulfate ......... 210 to 217.5 Cholesterol, R ............ 146 to 148
EthylaminobcnzDnte. . . . .. 88 to 90 Chrysoiclin, base ......... . 117
Etllyl morphine hydro- Cinchophen .............. 213 to 216
chloride .............. .. 123(d.) Cotarnine periodide ....... 142 to 144
Guaiacol. ............... . (a,) 28 Coumarin............ .... 67 to G9
Histamine phospllate ..... . (a.) 130 Dimethyla,minoazobenzcnc t

Homatropine hydrobro- R ..................... 115 to 117

mid.... . . . . . . . . . . . . . . . (a.) 212(d.) p-Dimethyll1minon,zoben-
Indofol'ln. . . .. .. . . . . . . . . . (n.) 115 zene, R ............... , 73 to 75
Isatin, R ................. 198 to 201 Ephedrine l,ydrnchlol'ic)e .. 214 to 220
Lurd ... " . . . . .. . . . . . . . .. 36 to 42 Ethyl carbamate ........ " 48 to 50
Menthol.. . . . . . . . . . . . . . .. 41 to 13 Guaiac....... . .. ... . .... 85 to 90
Methenamine. . . . . . . . . . . . (a.) 263 Guaiacol carbonate. . . . . . .. 8(j to 88
Neoeinchophen ......... ,. (n.l.t.) 74 Pnpaverine ............... 145 to 147
Oil, theobroma ......... " 30 to 35 Phenylhydrazine hydro-
Paraffin. . . . . . . . . . . . . . . . .. 50 to 57 chloride, R ............. 240 to 243
Petrolatum........... .... 38 to 54 Pilocarpine hydrochloride., 195 to 198
Phenacaine ............... 195 to 197 Salicin .................. , 199 to 202
Phenacaine hydrochloride. (n.J.t.) 189 Sulfonmethane ........... 124 to 126

(8.. ) = about. (n.1.t.) = not less than, (d.) = with decomposition. R. = reagent.
* Special method. s = sublimes.
 MELTING, CONGEALING, AND BOnING POINTS 231

4. What PCl' cent error would be introduced in the determination of the

melting point of salicylic acid as found if uncorrected?
6. Given two samples which show the same melting points; how might
you determine whether or not they were the same substance?

Congealing Point.-The congealing or solidifying point is a

physical constant characteristic of certain liquids, sunh as oil
of anise, oil of eucalyptus, and the fatty acids of fixed oils,
which may serve as a criterion of purity and quality just as the
melting point of a solid does, since the presence of impurities
also causes a depression of the congealing point. The official
method for the determination of congealing points is as follows:
"Unless otherwise directed, about 10 cc. of thc liquid to be
tested is introduced into a dry test tube of from 18 to 20 mm.
internal diameter, which is then cooled in water or a suitable
freezing mixture, the temperature of which should be about 5
lower than the supposed congealing point of the liquid. To
induce crystallization, rub the walls of the tube with a standard
thermometer, or add a small crystal of the substance being tested.
By alternate immersion of the tube in the bath, or removal from
the bath and constant stirring with the thermometer, the tem-
perature is so adjusted that the greater part of the liqlud gradually
congeals. The highest temperature remaining constant for a short
time elUTing the congelation is defined as the congealing point."
The solidification temperature of fatty acids is commonly
referred to as the titer. The official procedure for the determina-
tion of the solidifica.tion of fatty acids is illustrated in the follow-
ing exercise.

Exercise 76
Object.-Determination of the Solidification Temperature of
the Fatty Acids of Cottonseed Oil.
Materials Required.-50 ce. of cottonseed oil.
25 Gm. of potassium hydroxide.
100 ce. of glycerin.
15 ec. of alcohol.
"A standard thermometer meeting the following specifications: The
thermometer must have a zero mark, graduations of O.lC. between 10 and
600., and auxiliary reservoirs at the upper end and between the 0 and 10
marks. The cavity of the capillary tube between the 0 and 10 marks Illust
be at least 1 em. below the 10 mark, which must be from 3 to 4 cm. above
232 QU,<lN1'I7'..lTIVE PIIARkJACEUTICAL CHEMISTRY

the bulb, the totullength of the thermometer being about 38 em. The hulh
should he about 3 em. long and 6 mm. in diameter, the scale should he
etched on, and the graduations clear-cut and distinct. The thermometer
should be made of the best thermometric glass and thoroughly annealed, so
that scale errors 'vill not incl'case after continued heating."
Procedure.-l. "Heat 75 ec. of glycerin-potassium hydroxide solution
(made by dissolving 25 Gm. of potassium hydroxide in 100 cc. of glycerin)
to 150 0. in an 800-cc. beaker and add 50 cc. of the clarified fat, melted if
0

necessary. Heat the mixture for fifteen minutes with frequent stirring, but
do not ::Uow the temperature to rise above 1500. When saponification is
complete, the mixture is homogeneous, with no particles clinging to the
beaker at the meniscuR."
The oil is saponified by the KOI-I with the formation of the
potassiulll salts of the fatty acids and glycerin. See Saponifica-
tion Value (page 350) for the reactions that occur.
2. "Pour the soap into 500 ce. of nC[1rly hailing distilled water in an
800-co. beaker or o[1sserole, add slowly 50 ce. of dilute sulfuric acid (made
by adding 1 volume of sulfurie acid to 3 volumes of distilled wat81;), and
heat the solution with frequent stirring, until the fatty aeids separate
cleanly liS a transparent layer. Wash the acids with boiling water until free
from sulfuric acid, collect them ill a small beaker and place on a boiling
water bath or steam bath until the w[\ter has settled and the fatty acids are
clear. Allow the acids to cool, melt and filter into a dry beaker while hot,
and dry for twenty minutes at 100 0. 0

"Place 3 cc. of the dry acids in a test tube and add 15 ec. of alcohol. Heat
the solution to boiling and add an equal volume of ammonia water. A clear
solution should result."

The fatty acids are liberated from their potassium soaps by

the sulfuric acid. They are washed to remove potassium sulfate,
excess sulfuric acid, and glycerin and dried to remove the water.
If saponification is incomplete, the acids will be contaminated
with ullsaponified oil which would alter the congea.Iing point of
the acids. A turbidity in the alcohol solution indicates incom-
plete saponification since the oil is insoluble in the diluted alcohol
and the ammonium soap is soluble.
3. "0001 the dry, filtered acids to from 15 to 20 degrees above the expeet,ed
reading, and transfer to a glass tube 25 mIn. in diameter and 100 mm. in
length, the gl!1sS being 1 mm. in thickness. By means of a perforated cork
fasten the tube in a wide-mouth bottle of clear glass, approximately 70 mm.
in diameter and 150 mm. in height. Suspend the standard thermometer
in the melted acids so that it will serve as a stirrer, cooling if necessary, and
stir the mass slowly until the mercury remains stationary for thirty seconds.
 MELTING, CONGEALING, liND BOlLING POINT.S 233

Then allow the thermometer to hang quietly, the bulb in the center of the
acids !tnd observe the rise of the mercury colullln. The highest point to
whieh it rises is the Solidifying Point of the fatty acid"."
'Vhen the melted acids solidify, the "latent heat of fusion" is
liberated, and a rise in temperature OCCUl'R which remains con-
stant for a short time. The fatty acids give a distinct rise in
temperature when they solidify, whereas the fats and fatty oils
are usually characterized by the fact that they yield a constant
temperature which in some cases is variable when they solidify.
Questions and Problems
1. The U.S.P. requires that the congealing point of stearic acid should not
be below 54C. Why?
2. The determination of the congealing point of oil of eucalyptus con-
stitutes the official assay process. How does this determinatiou serve as Il.
measure of the purity of the oil?
TABLE XXXII.-OFFICIAL SUBSTANCES WITH CONGEALING POINT
REQUIREMENTS
Congealing
Subst!Lnce Point, cC.
U.S.P.
Acid, glacial acetic ........................ . (n.l.t.) 145
Acid, oleic ............................... . (n.m.t.) 10
Acid, stearic ............................. . (n.U.) 54
Bcuzene, R .............................. . (n.l.t.) 5.2
Euc!Llyptol. .............................. . (n.l.t.) 0
Fatty !Lcids from
oil, almond, expre~sed ................... . 9 to 12
oil, cottonseed ......................... . 28 to 35
oil, olive .............................. . 17 to 26
oil, theobroma ................. " ...... . 45 to 50
soap, hard .............. , .............. . 18 to 23
sodium stearate ....... , ................ . (n.l.t.) M
zinc s {"carate ........................... . (n.l.t.) 54
Oil of anise .............................. . (!l.l.t.) 15
Oil of eucalyptus ......................... . (n.Lt.) 15.4
Oil of fennel. ............................ . (n.l.t.) 3
Paraldehyde ............................. . (!l.l.t.) 11
PhenoL .... , ............................ . (n.Lt.) 39
Phenylhydrazine, R ....................... . (n.l.t.) 16
Suet, prepared ........................... . 37 to 40
N.F.
Anethol. ......................... , . . . . . .. 20 to 21
o-Toluidine, R .......................... " (n.m.t.) 20
(n.m.t.) = not more than. (n.Lt.) = not less than. R. = reagent.
234 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

Boiling and Distilling Points.-One of the characteristic

properties of a liquid is its boiling point. If a substance consists
of a single compound, it is generally considered to be pure when
it distils n,t a constant boiling point, under constant pressure.
In like manner, the purity of a mixture of substances, such as
11 volatile oil, may be judged in part by the range of temperature

at which the substance distils.

FIG. 27.-Apparatus for boiling and distilling point determinations.

The boiling point of any liquid may be defined as that tempera-

ture at which the vapor pressure of the liquid is equal to the atmos-
pheric pressure. Wh(~re a given temperature is stated as the
boiling point of a liquid, the inference is that the corresponding
pressure is standard atmospheric pressure, 760 mm., unless
otherwise given.
In the Pharmacopoeia, the boiling point is defined as that range
of temperature within which at least 95 per cent, by volume, of
the substance distils when treated in a definite manner.
The apparatus to be used (Fig. 27) and its construction are as
follows:
 MEL'l'ING, CONGEALING, AND BOILING POINTS 235

A distilling bulb of from 50 to 60 cc. capacity to the lower part

of the neck; the length of the neck should be from 10 to 12 cm.
and its internal diameter from 14 to 16 mm. The outlet tube
of from 4 to 5 mm. internal diameter should be attached to the
neck at approximately its mid-point, forming an angle of
70 to 75 dog. with the lower portion of the neck.
A straight glass condenser with a water jacket from 40 to 60 cm.
in length; the distance from the upper end of the water jacket to
the neck of the bulb being from 18 to 25 cm.
In order to avoid the necessity for an emergent stem correction,
it is advisable that an accurately standardized thermometer of the
Anschutz type be used. If such a thermometer is not available,
a thermometer used for melting points Illay be employed, apply-
ing the emergent stem correction as described under "Melting
Points,)) the length of the emergent stem being measured, in
degrees, from the lower end of the stopper to the highest point
of the boiling temperature. When placed in position, the lower
end of the thermometer should be from 2 to 3.5 cm. below the
center of the orifice of the outlet tube.
An asbestos board of from 12 to 15 cm. square and from 3 to 5
mm. in thickness, having a circular perforation, located centrally,
for the reception of the bulb. The edge of the asbestos around
the pcrforation should fit closely to the bulb when the latter is
set into it. The size of the perforation should be such that when
the bulb is set into it, the portion of the bulb below the upper
surface of the asbestos will have a capacity of from 3 to 4 cc.

Exercise 77
Object.-To Determine the Boiling Point of Carbon Tetra-
chloride.
Materials Required.-25 cc. of carbon tetrachloride.
A distillation apparatus as illustrated.
Procedure.-Place the asbestos board on a tripod or other suitable
support. Introduce into the distilling bulb 25 cc. of the liquid to be tested,
insert the thermometer, stand the bulb in an upright position in the per-
foration of the asbestos board, and connect it with the condenser. Then
distil the liquid at the rate of 1 cc. for each 15 to 20 sec., noting the tempera-
ture as soon as 5 drops of the liquid ha.ve distilled into the receiver and when
the last liquid evaporates from the bottom of the flask or when the specified
percentage has distilled over. Correct the reading for any variation in the
236 QUANTI1'ATIVE PHARMACEUTICAL CHEMISTRY

barometric pressure from t.he norl11al (760 mill.) by aliowing.O.loC. for each
2.7 mm., adding if the pressure is lower, and subt.racting if higher than
760 mm.
The temperature obtained when a thermometer is immersed
in a boiling liquid is usually greater than the true boiling point
of the liquid, because the vapor bubbles form below the surface
of the liquid and eonseqnently are at a pressure greater than
atmospheric. There is also an appreciable error due to super-
heating of the liquid, in this case. When the thermometer is
suspended in the vapor of the liquid, however, the bulb becomes
coated with a thin film of liquid in contact with its vapor, and
superheating is avoided.
It is often best to run a preliminary distillation, of a 25 cc.
portion of liquid, prior to the boiling point determination, in
order to ascertain the regulation of heating necessary to cause
the entire portion of volatile liquid to distil at the prescribed
rate. Superheating of the liquid and consequent bumping can
be prevented by placing a few small pieces of porous plate in the
distillation flask. The correction for the barometric pressure
may be made simply as illustrated in the following example:
If the range of temperature within which 95 per cent of glacial
acetic acid distils is 116.5 to 116.9C. at 733 mm., the corrected
temperatmcs would be 116.5 +
0.1 ( 7602.7
- 733) = n7.5, and

116.9 + 0.lC 60
2~ 733) = 117.9. The.boiling point of the acid
Rhould then be reported as b.P.m = 117.5 to 117.9C. cor., the
subscript 733 indicating the barometric pressure at which the
determination was made, and the abbreviation" cor." indicating
that the boiling point as reported has been corrected to standard
pressure. If a long-stemmed thermometer is used in the deter-
mination, a further correction for the emergent stem must be
made, as explained under Melting Point, page 229.
A second method is given in the Phn.rmacopoeia for use with
liquids for whieh the permissible range in boiling temperature
exceeds 5C. '1'ho following is tl description of the apparatus
required and the procedure to be followed:
A 200 cc. distilling bulb with an outlet tube at approximately
the center of the neck, forming an angle of from 70 to 75 deg.,
with the lower end of the neck. The length of the neck is from
 MELTING, CONGEALING, AND BOILING POINTS 237

10 to 12 cm. and its inside diameter is from 18 to 24 mm. The

length of the outlet tube is from 10 to 12 cm. and its inside
diameter is from 5 to 6 mm.
A straight glass condenser, a thermometer, and an asbestos
board are required as in Method 1. The diameter of the perfora-
tion in the asbestos board is 25 mm.
Place the distilling bulb in an upright position in the perfora-
tion in the asbestos board and connect it with the condenser.
The outlet tube is to extend from 25 to 35 mm. into the condenser
beyond the connecting stopper.
Measure 100 cc. of the liquid to be tested, using a cylinder
having 1 ce. graduations. Note the temperature of the liquid and
transfer it as completely as possible to the distilling bulb. Use
this cylinder as the receiver for the distillate without rinsing out
any of the adhering liquid. Insert the thermometer through a
stopper in the position described in Method I and heat with a
Bunsen burner protected from drafts until the liquid begins
to boil, then distil at the rate of from 4 to 5 ce. pel' minute. The
temperature when 5 drops of the liquid have distilled into the
receiver is the minimum distillation temperature, and unless
otherwise specified, the maximum distillation temperature is the
temperature at which 95 pOI' cent of the liquid has distilled over.
Bring the distillate to the same temperature at which the liquid
was originally measured, and note the volume. Correct the
temperature reading for barometric pressure and apply the emer-
gent stem correction as described in Method 1.
Liquids which begin to distil below 80 D C. are cooled to 10 or
15C. before measuring the 100 ce. for the test. The end of the
condensing tube is fitted with an adapter bent at a suitable angle
and the end of the adapter is passed through a cork inserted into
the receiving cylinder. The cork has a small perforation to
permit the exit of air. The receiving cylinder is kept immersed
in ice to within 2.5 cm. of its height during the distillation.
When boiling points are given with a range instead of with it
single figure, the figures are to be regarded as inclusive. The
boiling points of such substances as acetone, ether, ethyl oxide,
alcohol, glacial acetic acid, and tetrachlorethylene should be
regarded as informative rather than as criteria of identity and
purity since only approximate boiling points are given. Definite
requirements relative to the per ~ent of a substance which
238 QUliN'l'ITATIVE PHARMACEUTICAL CHEMISTRY

should distil within a certain rangc of temperature and above or

below a ccrtain temperature are given in the Pharmacopoeia and
National Formulary as follows: 85 per cent of creosote from
creOlolote carbonate distils at 200 to 2200., 90 per cent of creosote
distils at 203 to 2200., 90 per cent of cresol distils at 195 to
2050., 90 per cent of oil of turpentine and of rectified oil of tur-
pentine distils at 154 to 1700., and 85 per cent of liquid guaiacol
distils at 200 to 2100. It is required 'that the volatile oil
separated from copaiba by stcam distillation should not boil
below 2500., that less than 10 per cent of oil of dwarf pine needles
distils below 165C., and that the boiling point of liquefied phenol
should not rise above 1820. when distilled.
Those official substances for which a dcfinite range of boiling
points is spec.:ified and within which range 95 pel' cent of the
substance should distil are given in the following table.
TABLE XXXIII.-OFFICIAL SUBS'rANCES WI'l'H BOILING-POINT RANGE
Substance Boiling Point, C.
U.S.P.
Amyl alcohol, R ........ " ............. . 128 to 132
Aniline, R ............................ . 183 to 186
Benzene, R ........................... . 79.5 to 81
Benzin, purified petroleum .............. . 35 to 80
Butyl alcohol, R. ...................... . 116 to 118
Carbon disulfide, R .................... . 76 to 78
Carbon etrachlol'ide .................... . 76 to 78
Ethyl a tetate, R ...................... . 76 to 77.5
Ethyl chcloride ........................ . 12 to 13
Eucalyptol ............................ . 174 to 177
Eugenol. ............. '" ... " ........ . 250 to 255
Furfural, R, ........ , .. " ............. . 150 to 152
Guaiaeol, solid ......................... . 204 to 206
Kerosene, R ......................... : .. 180 to 300
Methallol, R ....................... " .. 64 to 67
Methyl salicyltLtC ....... " ............. . 219 to 224
Oil of lllllstard, volatile ................. . 148 to 154
Paraldehyde ........................... 120 to 125
Terebene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160 to 172
Toluene, R ............................ . llO to 111
Xylene, R ............................. . 137 to 140
N.F.
Ethylltcetate ........................ " 75 to 77
Anethol. ........ '............... '. : ..... " 234 to 237
R. = reagent
 MELTING, CONGEALING, AND BOILING POINTS 239

Questions and Problems

1. Define the terms boiling ]loint and dislilling ]Joint.
2. How is the distillation range of the cresol and of the creosote in creosote
carbonate directed to be determined in the Pharmacopoeia?
3. If ether contained alcohol as an impurity, would the boiling point of
the ether be mised or lowered?
4. A sample of carbon tetrachloride is found to distil between 76.8 and
77.g c C. at 664 mm. Calculate the correctccl distilling range of the carbon
tetrachloride.
6. Consult the appended table and list those substances the boiling points
of which should be determined by method II of the U.S.P.
6. Which is more pure as indicated by the required boiling point ranges,
ethyl acetate of the N.F. or ethyl acetate reagent of the U.S.P.?
 CHAPTER XIV

REFRACTOMETRIC MEASUREMENTS
The index of refraction is a physical constant frequently made
use of in the determination of the identity and purity of drug and
food products. In many cases, it may be used to determine
quantitatively the strength and purity of solutions or the propor-
tions in which liquids are mixedj e.g., the percentage of sugar in
syrup can bc estimated directly from the refractivity of the solu-
tion, and the percentage of alcohol in water can also be determined
in this way. Although the refractive index is a constant charac-
teristic of many substances such as fats, fatty
oils, waxes, sugars, organic solvents, etc., it is
applied almost exclusively in the official stand-
ards as a criterion of the purity of volatile oils.
Index of Refraction.-When a ray of mono-
chromatic light passes from one transparent
substance into another of different optical
density, it is deflected or refracted except
FIG. 28.-Refrac- when it enters perpendicularly to the surface
tioll of light.
of con t act between the substances. The
extent and direction of the deflection are dependent upon the
difference between the densities of the two substances. The
angle between the ray in the first medium and a perpendicular to
the dividing surface is termed the angle of incidence i, and the
. corresponding angle in the second medium is called the angle of
refraction r. The sin of i and the sin of 7' are directly proportional
to the velocities of light in the two media. The ratio sin i/sin r
is termed the index of refraction. Thus, the velocities of light
in ail' and in water are in the ratio of about 4 to 3. The index of
refraction of water with respect to ail' is therefore about 1.333.
In Fig. 28, I is the less dense medium and II the more dense
medium. A ray of light passing obliquely from I to II will be
deflected so that the angle of refraction r will be less than the
240
 REPRAC'TOME'l'RIC ME'A8UREMENTS 241

angle of incidence i, and according to the law of refraction, the

relationship between these two angles will be sin i/sin r = N /n
where n is the index of refraction of the less dem:e and N the index
of refraction of the more dense medium. When the angle i is
increased, the angle l' also is increURcd and attainR its maximum
value when the angle of incidence enters horizontally at 90 deg.,
termed grazing incidence, or at a right angle. In the latter case,
since the sin of 90 deg. = 1, the above eqnation becomes IJsin
l' = N /n or sin l' = niN. The angle 90 dog. is known as the
critical angle and the ray of incident light passing from one
mediulll to another at this angle is known as the critical ray.
If the angle r is increased to a value at which i becomes equal to
90 deg., the beam of light \vill no longer pass from the first
medium to the second but will travel through the first medium to
the dividing surface at an angle of 90 dog. with the perpendicular
to the surface. If r is smaller than this particular value, however,
the ray of light will pass on through the second medium, and, if
greater, all light will be reflected from the surface back into the
first mediulll. The critical ray is thus seen to make a boundary
between the reflected and refracted light. This furnishes the
basis for the observation of the refractive end point used in many
refractometers.
Refractometers.-Indices of refraction may be determined by
means of a polarizing microscope or refractometer. Refractom-
eters are of several types. Some of these give figures on an
arbitrary scale, the refractive iildex being found by reference to a
table furnished with the instrument. The principal instruments
in use are (1) the Pulfrich refractometer, an instrument adapted
for use with liquids and solids of all degrees of refractive power,
used chiefly for purely scientific investigations; (2) the Zeiss'
butyro-refractometer, designed especially for the examination
of fixed oils and fats; (3) the Zeiss immersion refractometer,
especially constructed for the examination of aqueous, oily, or
alcoholic liquids when a large amount of sample is available; (4)
the Abbe refractometer, an instrument designed for the examina-
tion of liquids, solids, and plastic bodies. Since this last instru-
ment is best suited for the examination of all of the official sub-
stances for which refractive indices are given, it is described in
considerable detail.
 242 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The A.bbe ReJractometel'.-BecauflB of its simplicity. ease of

manipulation, and the small amount of material required in its
use, the Abbe refractometer is extemdvely used in pharmaceutical
analysis. The wide range of indices measurable with this instru-
ment and the fact that it can be used with ordinary white light
make it capable of more general application than any of the
other refractometers mentioned above. When measuring the
refractive index of liquids, only a few drops) sufficient to form a

FIG. 29.-Abbli refractometer.

film, need be placed between the prisms of the instrument.

This is a distinct advantage over the other forms of refractom-
eters, for frequently only a small quantity of the substance to
be examined is available, and if the liqnid is colored or turbid,
the thinness of the film permits a determination of the index of
refraction without difficulty.
The Abbe refractometer (Figs. 29 and 30) consists of four
essential parts, namely: (1) a telescope E provided with an
objective, an eyepiece, and a cross-hair disk. The purpose of the
telescope is to form .an image of the border line of total reflection
 REFRACTOMETRIC MEASUREMENTS 243

at the intersection of the cross-hairs where the image may be

observed. (2) Two similar glass prisms (Abbe prisms) of high
refractive index cemented into hollow, water-jacketed, mounting
cases, so constructed that when temperature control is required,
water can be circulated about the prisms. The exposed surfaces
of the upper prism B are highly
polished, while the hypotenuse sur-
face of the lower prism A, which
serves solely for the purpose of illumi-
nation, is finely ground. The lower
prism mount is hinged to the upper
prism mount and the latter iH rigidly
attached to the index arm or alidade.
,Vhen the two prisms are clamped
together, a space of from 0.1 to 0.15
mm. separates their surfaces. (3) The
Bector H is a metal arm to which the
telescope is rigidly attached. One
end of the sector is attached to the
upright of the base in such a manner
that the whole sector can bo rotated
about an axis, which coincides with
the axis of rotation of the Abbe
prism, so that various conditions of
. illumination can be accommodated.
On the upper end of the sector a scale
G is mounted which is graduated
directly in terms of refractive index FIG. aO.-Path of light, etc.,
through Abbe refractometer.
of the D line (sodium light) at 20C.
An alidade or movable arm J rests against the sector, but it
can be rotated' as a unit with the Abbe prism independently
of the sector. rEhe upper end of the alidade bears an index
mark which moves along the edge of the sector scale when
the arm is rotated. A magnifier F is attached to the alidade
in most instruments to facilitate the reading of the index
scale. A screw M is used to make fine adjustments of the
border line at the cross-hairs. (4) Compensating prisms are
used to correct for the dispersion of light. Thus, when white
light is used in the place of sodium light, the beam is split
244 QUANTITA'I'lVE PHA.RMACEUTICAL CHEMISTRY

up into colored beams, siuce short wave lengths of light are

refracted to a greater extent than arc long wave lengths.
Unless this dispersion is compensated for in some way, the
border line becomes a broad, varicolored band which cannot
be brought into accurate coincidfmce with the cross-hairs of the
telescope. The compensator fUllctions to bring tlie various wave
lengths of light together again. It is placed between the upper
prism and the eyepiece and is rotated by means of the milled
head D.

FIG. 31.-Temperature control apparatus for Abbe refractometer. W.

Water supply. WR. Water-pressure l'egulator. S. Spiral heatCl'. B. Bunsen
burner. C. Water-pressure regulator. P. Inlet stopcock. R. Refractometer.

Regulation of Temperature.-Since the density of both the

prisms and the substance under investigation varies with thE:
temperature, it is necessary in refractivity meaSlHements of any
degree of accuracy to be able to maintain the prism and the
substance at constant temperature, In the Abbe refractometer,
this control of temperature is accolllplished by circulating water
from a thermostat 01' other suitable constant tempcrature device
about the jacketed prisms, the water entering the inlets I and L
passing out of the outlets 0, the temperature at whir.h the meas-
urement is made being obscrved on the thermometer T. A
temperature control device is illustrated in Fig. 31.
 REPRllC'1'OME'l'RIC MEilSUREj)[EN'1'S 245

Exercise 78

Object.-To Determine the Refractive Index of Oil of Orange.

Materials Required.-About 1 ce. of oil of or[l,uge.
A pipette.
An Abbe refl'llctol11etcr.
Procedure.-l. Rotu.te the prisms and telest,ope flO tlmt the lower prism
may be swuug open easily. Attach the test plate to the upper prism, by
means of the ll10110brom-Imp\lthalene or other liquid provided with the test
plate, by moistening the test pIa1.e with the liquid and pressing it ag:linst
the uppel' prism B. Focus the light by means of the lower prism, on the
test plate, rotate the prism by means of the alidade until the border line
of total reflection coincides with the cross-hairs in the tel('seope, and take
the ]'(;ading of the refrttcti ve index.

Th(~ refractive index of tlw test plate is etched upon it, and the
reading of the instrument should correspond to that given on this
plate. If the instrument is not in adjustment, a correction should
be applied to the readings obtained.
2. Remove tlle test plate, dean Lire llpper prism \VItI! cotton saturated
wit.h alcohol; damp the prisms tugeiJlC!l' looHcly, introduce 2 or 3 drops of
the volatile oil into the groove at the side of the prisms, and damp the
pri8m~ together firmly. AfljnRt the mirror K HO that the light is re.fleeted
upon the lower prism, and rotaUl the nJidatlc until the border line between
the light alit! clark halves of the field of view exaetly eoincides with the
cross-hairs of the teleAcope, .rotating the compensator prisms to obtain a
sharp uncolored border line if neeessary. Read the reimetive index: of
the oil directly from the gra([uated seetor sCllIn to the fourth deeimal place.
Move the alidade and again determine the refract.ive index unt.iI three
readings nrc obt:1ined, taldng the mean of the readings us the refmctive
index of Lhe oil. Note the temperature :Lt which the refractive index is read.

The readings on the graduated scale of the compensator drum

may be referred to tables of mean dispersion and c011version
supplied with the instrument. The refractive index of the oil
should be expressed as, e.g., N~oo = 1.4725 where N~oo denotes
the index of refraction for the D line (sodium light) measured
at 20C.
Questions and Problems
1. Define the terllls index of refraction, critical ray, and angle of incidence.
2. What advltnt,agcs docs t.he Abbe refractometer possess over the other
types of rcfractometers?
246 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

3. A l'ange is given for the refractive index of most of the official volatile
oils. Why?
4. How do variations in temperature affect the indices of refraction of
liquids?
o. Would you expect tlH1t oil of orange would have a greater or smaller
index of refraction at 25 than at 200. ? Why?
TABLE XXXIV.-OFFWIAL SUBS'l'ANCES WITH REQUIRED REFRACTIVE
INDICES

Temperature,
Substance 00. Official requirement

D.S.P.
Eucalyptol. ................... , , . 20 1. 455 to 1. 460
Oil of anise .......... , ......... , .. 20 1 . 5530 to 1. 5600
Oil of bitter almond ............ , . , 20 1 . 5428 to 1. 5439
Oil of chenopodium .. , ........... . 20 1 . 4723 to 1. 4790
Oil of cinnamon .. , ..... , ..... , ... . 20 1 .6020 to 1.6135
Oil of clove ... , .... , . , , .. , , , , .. , , . 20 1 . 5300 to 1. 5350
Oil of coriander, , .. , , . , , , , , , , , . , , . 20 1 , 4620 to 1, 4720
Oil of dwarf pine needles. , . , . , .... . 20 1. 4580 to 1. 4700
Oil of eucalyptus. , , ........ , , .... , 20 1 . 4600 to 1, 4690
Oil of fennel. ...... , ........ , .... . 20 1.5280 tu 1,5380
Oil of juniper. , . , .. , . , .. , ........ . 20 1.4780 to 1. 4840
Oil of lavender, , ...... , , , . , , , , ... . 20 1. 4590 to ] .4700
Oil of lemon, , , . , .. , .. , . , , , , . , .. , . 20 1,4742 to 1. 4755
Oil of mustard, volatile .... , . , ... , . 20 1 . 5268 to 1. 5280
Oil of myristic a ........... , . , .... . 20 1.4740 to 1. 4880
Oil of orange, . , . , .. , . , , ... , ..... . 20 1. 4723 to 1. 4737
Oil of peppermint ....... " , . , . , .. . 20 1. 4600 to 1. 4710
Oil of rose, , ............ , . , ...... . 30 1. 457 to 1. 463
Oil of rosemary .................. . 20 1. 4640 to 1. 4760
Oil of santa!. .................... . 20 1. 5000 to 1. 5100
Oil of sassafras .................. . 20 1 . 5250 to 1. 5350
Oil of spearmint ..... , . , " , ...... . 20 1 .4820 to 1,4900
Oil of theobroma, ..... , .. , .. , .... . 40 1.4537 to 1.4578
Oil of turpentine ................. . 20 1. 4680 to 1. 4780
N.F.
Anethol. ...... ' ....... , ......... , 25 1. 5580 to 1.5610
Oil of caraway ................... , 20 1. 484 to 1. 488
 CHAPTER XV

ROTATORY POWER

Many organic substances, such as certain constituents of the

volatile oils, alkaloids, and sugars, possess the power of rotating
the plane of polarizcd light when the latter is passed through
solutions containing them. Substances which possess this power
are said to be opt1'cally active and are designated as dextrorota-
tory when the direction of the rotation is toward the right and as
laevorotatory when it is toward the left.
The optical activity of many substances is a function of their
chemical constitution as well as of their concentration. COIl-
sequently, a determination of the rotatory power or lack of
rotatory power of a substance may serve as a means of establish-
ing both its identity and its purity~ In some cases, the meas-
urement of the optical activity of a substance may also give
some indication of its therapeutic value; e.g., the laevorotatory
alkaloid hyoscyamine is much more active as a mydriatic than
its optically inactive isomer atropine.
In a ray of o1'dinary light, the vibrations are transverse, i.e.,
they take place in a plane at right angles to the direction of
propagation, but the vibration direction is constantly changing.
In a ray of plane-polarized light, commonly termed polarized light,
the vibrations are also transverse, but they take place in only
one direction.
The polariscope and the polarizing microscope are used in the
determination of the optical activities of liquid and solid sub-
stances, the polarizing microscope being employed more often
for the examination of solid substances, while thc polariscope
is used primarily for the examination of liquids.
When a ray of ordinary light is passed through a specially
constructed prism made of Iceland spar, such as the Nicol's
prism, the emergent light wave vibrations take place in one
plane, and the light is said to be .plane-polarized. If another
, 247
248 QU.ilN1'I1'A1'IVE PHARMM'EU1'IC'AL CHEMISTRY

Nicol's prism is used to examine the plalle-polarizecllight, it will

be found that on rotating the lattcr prism, the field of view
appears alternately light and clark, tlH~ minimum brightness
following the maximum as the prism is rotated through an angle
of 90 cleg. The prism by means of which the light is polarized
is termed the polarizer, and the sceond prism, by means of which
the light is examined, is called the analy;zc1'.
If, when the field of view appears clark, as when the axes of the
two prisms are at right angles to each other, a tube similar to
that shown in Fig. 32 or 33 is filled with an optically active
liquid and placed between the polarizer and ~Ulalyzer, the field
will become light due to the rotatiol1. of the plane of polarization

FIG. 32.---Polariscope tube. lira. 33,-Polariscope tube

with jacket aud thermometer
well for constant temperature
control.

by the optically active substance. Tlw extent to which the plane

of polarization has been rotated can be determined by turning
the analyzer Ghrollgh a certain angle until the field becomes dark
again. When it is necessary to turn the analyzer to the right
(clockwise), the optically active substance is said to be dext1'Orota-
tory; when the analyzer must be turned to the left (counter-
clockwise), it is said to be laelJorotatol'Y,
It is possible to obtain a position at which the fielcl of view
becomes dark by rotation of the analyzer to either the right or
left, since in one complete rotation of the prism through 360 cleg.,
there are two positions of the analyzer, 180 deg. apart, at which
the field becomes dark, and, likewifle, two positions at which
there is a maximum of light, In determining the sign of activity
of a given substance, it is customary to take the direction in which
the rotation required to reproduce the dark field is l&,>s than
90 deg,
The angle of rotation of the polarized light on passage through
a substance varies directly as the length of the column of sub-
 ROTliTORY POWER 249
stance through which the light paSSflS. In homogeneons mixtures
or solutions, the angle of rotation depends upon the ~oncelltration
of the optically active :mbstanees present. ~Whcl1 more than one
optically active substance is present in a solution, the resultant
rotation will be the srtme as the SUIn of the sever:ll rot.ations of
each separate c:ollstituent in the RaHle volume of solution, pro-
vided that the substances do not react with each other. The
angle of rotation also varies with the wave length of the light
employed (the shorter the wave length the greater the augle of
rotation), with the temperature, and with the nature of the
substance. When all of t.hese fadm's are taken into aecount, the
results may be calculated in terms of specific rotation or specific
rotatory power.
The speeifie rotation of an optically active liquid is defined
as thc anglc of rotation in dC(frces through which the plane of polariza~
tion of monochromatic CD) light is rotated by passagc thro'ugh a
liquid containing 1 Gm. of actilJc 8ubstance in a volume of 1 ce.
determined in a tube havin(J a length of 1 don. Since the specific
gravity of liquids I1S well as the optical activity of many sub-
stances is influenced hy the temperature, it is necessary to indicate
the temperature at 'which the rotation and the specific gravity are
determined. The temperature specified in the two official
standards for the measurement of optical activity is generally
250., exeept in the case of sugar which is determined at 200.
by international agreement.
The caleulation of the speeific: rot(Ltory power of an optically
active liquid or solution of an optically active solid may be made
from the following general formulae:
1. For liquid substances [alb
. 100a
II. For solutIOns [aJb = lpd
lOOa
t
or [a ID = --r:c-
For calculating the specific rotation [Ic] using these formulas,
the determination of the following factors is necessary:
a = observed rotation in degrees of the liquid at temperature
t using a sodium light.
l = the length of the tube in d~cimeters.
250 QUANTITATTVE PHARMACEUTICAL CHEMISTRY

d = the specific gravity of the liquid or solution at the temper-

ature of observation.
p = concentration of solution expressed as the number of grams
of active substance in 100 Gm. of solution.
c = concentration of solution expressed as the number of
grams of active substance in 100 cc. of solution.
In expressing the specific rotation of a substance in solution
the concentration and solvent used must always be stated. The
oo
temperature should also be recorded; e.g., [ag = -38 0 means
that the specific rotation of the substance determined at 20C.
using sodium light is laevorotatory 38 deg., and [al~Oo = + 16
means that the substance is dextrorotatory 16 deg. when the
specific rotation is determined at 20C., using sodium light.
Polarimeters.-The instruments used to measure the optical
activity of liquid substances are called polaril3copes or polarimeters
i , , -----~I
... J::I'/~
I~F ~ IDE c:s::JD 1 ~)=I=:::::Q~=:Jq] ,Ie !:SIs A0 1~Rt
(]let
FIG. 34.-Diagram of the optical parts of a polarimeter.

and l3accharimeters. Polarimeters permit the angle of rotation to

be read off in degrees or fraetions of a degree of a cirele, while
saccharimeters are usually constructed and set to give readings
directly in terms of percentage of sugar. The so-called half-
shadow instruments known as the Laurent and the Schmidt and
Hansch polarimeters are most generally used in pharmaceutical
work.
In the polarimeter of Laurent and that of Schmidt and Hansch,
the optical parts may be represented diagrammatically as in Fig.
34. Monochromatic light (sodium light) from the source L,
passes through the lens A, which renders the rays of light parallel,
and then passes tlll'ough the polarizing priKm B, which renders
the light ray plane~polarized. It then passes through the obser-
vation tube 0, which contains the substance being examined,
and thence through the analyzer prism D. '1'he field of view is
observed through the telescope FE. The circular opening
of the tube carrying the polarizer is one-half covered by a t,hin
. quartz plate as illustrated in a', the thickness of which is so
adjusted that the light in passing through the plate is altered
 ROTATORY POWER 251

in phase by half a wave length but remains plane-polarized. In

this way, two beams of polarized light arc produced. If the
polarizer is rotated so that the plane of polarization forms an
angle X with the quartz plate, the planes of polarization will
also be inclined at an angle 2X. This is termed the half-shadow
angle. On rotating the analyzer, a position will be found at
which one beam of light will be completely, and the other only
partly, extinguished. One half of the field of view will then
appear dark while the other half will remain light. Upon rotating
the analyzer farther through the angle 2X, a second position will

FIG. 85.-A polarimeter.

be found at which the second beam of light will be extinguished

while the first beam becomes visible. At this position of the
analyzer, that half of the field which formerly appeared dark will
be light and formerly light will be dark. When the analyzer
occupies an intermediate position, a field of view of uniform
brightness will appear, and this is the position at which the
analyzer should be set.
The complete polarimeter is shown in Fig. 35. At the end S,
which is directed toward the source of illumination, is the lens.
The polarizing prism is at P, and it is connected with a lever
L, by means of which it can be rotated to alter the angle of half-
>1l,aclow. The observation tube is placed in the central part
,;1 the instrument and protected from extraneous light by a
hinged cover. The analyzer is placed at A, and it can be rotated,
252 QUANTITATIVE PHARMACEUTICAL CIlEMIS'l'RY

independrmtly of the graduated circle, by means of a screw. This

permits the correction of the zero point of the instrument. F
is a telescope: with an eyepiece by means of which the field of
view is observed. K is a graduated disk which can be made to
rotate with the analyzer and telescope past the fixed verniers n
011 the graduated eirele. Most instruments are provided with
Due or more magnifying lenses to increase the accuracy of
readings.
The official polarimetric determinations should be carried out
in a dark room using monochromatic light. Monochromatic
light can be obtained easily by introdueing a fused bead of sodium
ehloride on a platinum wire or gauze into the non-luminous flame
of a gas burner.
The observation tub~, in which the liquid to be examined is
placed, usually consists of a tube of thick glass with accurately
ground ends, dosed by eireultVr glass plates with parallel sides
which are pressed against the ends of the tube by meam; of screw
caps. Other polarimeter tubes are eonstrncted with a jacket
through which wat,er may be circulated t.o give a constant. t.emper-
ature. Since the unit of length in polarimetric measurements is 1
dem., the observation tubes are generally of 1 demo length or some
multiple or fraction thereof, c.g., 2.0, 0.5, aud 0.25 dClll. The
length of the tube used in any definite determination usually
depends upon the depth of eolor in the liquid and the magnitUde
of its rotatory power.
Readings made on a saccharimeter may be converted into the
angular degreeH uf a polarimeter by means of the following factors:
1 division of the Laurent sugar scale = 0.2167 clog. angular
rotation D.
1 division of the Ventzke or Schmidt ltnd Hiinseh sngar scale
= 0.3468 deg. angular rot,ation D.

Exercise 79

Object.-To Determine the Sp(~cific Rotation of SUerose.

Materials Required.-26 Gm. of oane sugar.
A 100 CG. volumetric flask calibrated at 200.
A polarimeter with 11 200 mm. observation tube.
PTocedure.~l. Adju~t and determine the zero point of the polariinetel".
 ROTATORY POlVER 253

The polarimeter should be set up so that the polarizer end is in

direct line with the bright sodium flame and about 4 to 6 ill. from
it. Place a 200 mm. observation tube filled with distilled water
in the tube holder between the polarizer and the analyzer and
focus the telescope eyepicce on the lille bisecting the field of
view, rotating the analyzer if necessary in order to obtain unequal
illumination of the two halves of the field. Now determine the
zero point by rotating the analyzer until equal illumination of the
two halves of the field is obtained. This position should be
approached several times from either side of the zero point,
readings being made from the verniers, and the mean of at least
four readings taken as the zero point. Since the zero point is
altered by the alteration of the angle of half-shadow, the lever
whieh rotates the polarizer must he set before the zero point is
determined. It is possible to rotate the analyzer in some instru-
ments without rotating the graduated disk, and it is possible,
in such cases, to adjust the instrument so that the zero point
correction is eliminated.
2. Dissolve 26 Gm. of Sllcrose, accurately weighed and prcviously dried
to constant, weight at 105C. in sufficient distilled water to make 100 ce. at
20C.

The sugar is dried to drive off moisture. The solution should

be made up in a volumetric flask calibrated at 20C., and the
final adjustment of the volume should be made at the same
temperature.
3. Fill a clean dry observation tube with the solution. Determine the
value of the allglc of rotation. Calculate the specific rotation of the sample
examined.

A polarimeter tube may be cleaned and dried easily by washing

it successively with water, alcohol, and ether. In the determina-
tion of the angle of rotation, a number of successive readings
should be taken as described nnder procedure 1, and their mean
taken as the angle of rotation. After correcting for the zero
point of the instrument, the specific rotation can be calculated
by the formulae given on page 249.
The specific rotation of the official sucrose must not be less
than +65.9 deg. when determined by the above method. Com-
254 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

pare the specific rotation of the sample eX!1mined with the official
requirement.
Questions and Problems
1. What is polarized light? How can it be obtained?
2. Show diagrammatically the essential parts of the polariscope used
and explain the function of each part.
3. What factors influence the. rotatory power of a substlLllce?
4. Define the term specific rotatory power.
TABLE XXXV.-OFFlCIAr, SUBSTANCES WITH REQUIREIJ ANGULAR ROTATION
Official Requirement
Substance a = Angulur rotation
U.S.P.
Copaiba, volatile oil from. . .. a = not less than _7
Methyl salicylate from gaul-
theria ................... , a = not ~nore than -1.5
Oil of anise ........ , ........ ' a = +1 to _2
Oil of bitter almond ......... a = inactive to +0 0 10'
Oil of cinnamon ............ , a = +1 to _10
Oil of clove ................ , a = not more than _10 10'
Oil of coriander. . . . . . . . . . . .. a = +8 to + 15 0
Oil oUennel. ...... , ........ a = +12 to +24 0
Oil of juniper. . . . . . . . . . . . . .. a = 0 to -15 0
Oil of lavender ..... , ......... a = -3 to _10 0

Oil of lemon ................ a = +57 to +65.6 0

Oil of dwarf pine needles. . . .. a = -5 to -12 0
Qil of myristica. . . . . . . . . . . .. a = + 10 to +30 0
Oil of orango ............... , a = +94 to +99 0
Oil of peppermint ........... a = -23 to -320
Oil of rose ......... , ....... a = 1 to 4 0
Oil of rosemary. . . . . . . . . . . .. a = 5 to + 10
Oil of santal ................ a = -15 to -20 0
Oil of sassafras .. " .......... a = +2 to +4
Oil of spearmint ............. a = -48 to -59 0
Terehene. . . . . . . . . . . . . . . . . .. a = not more than +0.3
N.F.
Anethol. . . . . . . . . . . . . . . . . . .. a = 0 to +0.08
Oil of bergamot .... , ..... '" a = +8 to +24
Oil of bitter orange .......... a = +88 to +98 0
Oil of caraway. . . . . . . . . . . . .. a = +70 to +80
Oil of cardamom. . . . . . . . . . .. a = +22 to +44 0
Oil of myrcia ........... " .,. a = not. more than _3
Oil of orange flowers. . . . . . . .. a = +1 0 30' to +9 8'
Oil of pi me uta .............. a = 0 to _4 0
Oil of thyme ................ a = not more than 4"
Salioin .. , . . . . . . . . . . . . . . . .. a = 62 to 67
 ROTATORY POWER 255
5. Explain what the following terms mean: (1) [alif = +4.210; (2)
[am" = _10"; (3) dextrorotatory; (4) optically inactive.
6. The optical rotation of a sample of oil of ehenopodiUIn determined in a
100 mIn. tube and having a density of 0.9750 is found to be -12.25 dog.
at 210. Calculate the speeific rotation of the oil.
7. A solution containing 10 Gm. of sugar in 100 ec. was found to exhibit
an angular rotation of +11.5 dog. when determined in a 100 lllm. observa-
tion tube. Calculate the specific rotation of the sucrose.

The official substances with their optical activity requirements

may be classified as follows.:
1. Those which are required to be optically inactive.
U.S.P. Atropine, eucalyptol, eugenol, mcthyl salicylate
(synthetic and from Betula), vanillin, and volatile oil of mustard.
N.F. Anethol, arecoline hyclrobromide, and papaverine
hydrochloride.
2. rrhoRB which are required to be laevorotatory.
U.S.P. Cocaine, codeine, colchicine, honey, menthol,
quinine, quinine bisulfate, quinine sulfate, and santonin.
N.F. Cinchonidine sulfate.
TABLE XXXVr.-OFFICIAL SUBSTANCES WITH REQUIRED SPECIFIC ROTATION

Cone en-
Solvent
Substance
used
trationin [aln = specific rotation
100 ce.

U.S.P.
Camphor, natural Alcohol 10 Gm. [all!'" = +41 to +42 0
Dextrose ......... Water 10 Gm. [a]W = +52.5 to +53
Ephedrine hydro-
chloride ..... , . , Water 5 GIll. [alJr = -33 to -35.5"
Ephedrinesulfate .. Water 5 Gill. [alto" = -29.5 to _320
Epinephrine ...... 0.5 N HCl 5 Gm. [alto" == -50 to -53.5
Oil, chaulmoogra .. Chloroform 10 Gm. [alt" "" +48 to +60
Ethyl chaulmoo-
grate ........ , .. Chloroform 50 cc. [ant = nob less than +44.5
Laetose ..... ,., ... Water 10 GIll. [an:" = +52.2 to +52.5
Scopolamine hy-
drobromide .. , .. Water 5Gm. [aHt = -22 to -25.75
Sucrose .......... Water 26 Gill. [aut = not less than +65. go
N.F.
Ephedrine sulfate,
solution of ...... Water 3Gm. [altaO = -28 to -30
256 QUAN1'ITA'l'IVE PHARi}fACEUTICAL CIJEMl81'RY

3. Those which are required to be dextrorotatory.

U.S.P. Quinidine sulfate.
N.F. Cinchonine sulfate.
4. Those for which the angular rotation at 25C. determined
in a 100 mm. observation tube is given in Table XXXV, page 254.
5. Those for which the specific rotation is given in Table
XXXVI, page 255.
All of the official determinations of specific rotation are per-
formed in a 200 nun. tube, except that of scopolamine hydro-
bromide in which a 100 mill. tube is used.
The measurement of rotatory power is made the basis for the
assay for dextrose in the alllpuls of dextrose, ampuls of dextrose
and sodiulll chloride, and isotonic solution of dextrose and
sodium chloride of the N.F.
 CHAPTER XVI

VISCOSITY MEASUREMENTS
The viscosity of a fluid substance is constant for any given
temperature and is a measurable characteristic of the substance.
It is used chiefly as an index of the composition and lubri-
cating value of oils.. The viscosities of solutions and liquid
mixtures often vary with their concentration and composition,
however, this property may be used, in many cases, as a rapid
means of analysis. The U.S.P. utilizes the viscosity as a means
of standardizing liquid petrolatum which is employed therapeu-
tically as an intestinal lubricant.
In the U .S.P., viscosity is defined as "a term used to denote
the relative degree of fluidity of a liquid." This definition is
confusing, since fluidity is the reciprocal of viscosity; i.e., fluidity
= l/viscosity. Fluidity may be regarded as a measure of the
tendency of a liquid to flow, whereas viscosity is a mf.asure of the
resistance which a liquid exerts against the tendency to flow.
Viscosity may be defined as the force of friction which tends
to retard movement in a fluid body. The unit of absolute viscosity
is defined as the tangential force required to move a unit area of
plane surface within the liquid with unit velocity relative to another
parallel unit area of plane surface one unit distant from the former
surface, where the unit of force is the dyne (the force necessary
to produce an acceleration of 1 cm. per second per second on a mass
of 1 Gm:), the unit of velocity is the centimeter per second, the unit
of area is the square centimeter, and the unit of distance is the centi-
mete1. This unit of absolute viscosity is known as the poise and
it is generally expressed as dyne-second per square centimeter.
The poise is a relatively large unit, and the one-hundredth part
of the poise, or a centipoise, is commonly employed as the unit of
absolute viscosity.
Instead of giving results in terms of absolute viscosity, most
methods of getermination give the- relative viscosity; i.p,., the
257
258 QUANTITA.TIYE PHARMACEUTICAL CHEMISTRY

viscosity of the liquid compared to that of some standard of

known viscosity, such as water under similar conditions. Since
the relative viscosities obtained by determinations with different
types of apparatus are not the same, the kinematic viscosity,
which is the ratio of the absolute viscosity in poises to the density,
has been adopted in the U.S.P. as the method of expressing
viscosity. As the viscosity of a liquid changes very rapidly
with the temperature, decreasing about 2 per cent for each
degree rise'in temperatmc, it is important to know the exact
temperature of the liquid at the time of measurement and to
use an instrument provided with some means whereby the sample
under examination may be main-
tained at constant temperature.
Apparatus.-In the official deter-
mination either the Eng 1e r 0 r
Saybolt \riscosimeter may be em-
ployed, and the results obtained
with these instruments may be con-
verted into the kinematic viscosity
by the use of a conversion table.
Both of these instruments are con-
structed to meal:lure the tim e
required for a certain amount of
fluid to pass through a capillary
tube. Whcn the Engler instrument
is used, the viscosity is expressed in
terms of "Engler degrees," obtained
FIG. 36.-Saybolt universal vis- by dividing the time of discharge
cosirnctcr.
for the liquid being tested by the
time of discharge of the same volume of water at 20C. In this
country, the Saybolt universal viscosimeter is most commonly
employed ill the determination of the viscosity of lubricating
oils. The results obtained with this instrument are expressed
as the time in seconds required for the discharge of the liquid
being tested.
The SayboZt universal viscosimeter (Fig. 36) consists of a metallic
cup of about 80 cc. capa<Jity to contain the liquid under examina-
tion with an outlet in the bottom closed by a cork stopper.
This cup is surrounded by a jacket which serves as a constant
temperature bath. Most instruments are provided with devices
 VISCOSITY MEASUREMENTS 259
for stirring and heating the bath, with a thermometer to measure
the temperature, and with a receiving flask graduated to contain
60 cc. at 20C.; a stop watch with which the time of discharge
of the liquid may be measured is usually a part of the standard
equipment supplied with the instrument, but any stop watch
lllay be used.
Exercise 80
Object.-:-To Determine the Kinematic Viscosity of Liquid
Petrolatulll.
Materials Required.-A Saybolt viscosimeter.
A stop watch.
100 cc. of liquid petrolatum.
Procedure.-l, Assemble the viscosimeter, fill the cOllstant tempera-
ture bath with water, and bring the ternperatUl'e of the latter to about 40C.
Tightly stoppel' the outlet tube, and pour the sample of petrolntum into
the oil cup until the level is above the rim of the main cup. When the
temperature of the oil sample is 37.8C., pipette the excess oil from the
overflow cup until thc level in it is below the rim of the main cup.

The water in the temperature bath should be stirred continu-

ally to insure an even distribution of temperature throughout
the sample of oil.
2. When the thermometer in the oil GUp registers exactly 37.8C., remove
the cork 1'om the outlet tube, starting the stop watch at the same instant,
and collect the effluent oil in the flask graduated to hold 60 cc. Whcll the
level of the oil in the flask reaches the 60 cc. graduation mark, stop the
watch.

The time in seconds required for the efflux of 60 ce. of the liquid
petrolatum is called the Saybolt universal viscosity.
The U.S.P. requires that liquid petrolatum have a kinematic
viscosity of not more than 0.370 at 37.80. (lOOF.). This value
is equivalent to a Saybolt universal viscosity of 170.
Questions and Problems
1. Consult the viscosity conversion table (U.S.P., page 477) and expreSH
the results obtained in the above exercise in terms of kinematic viscosity
and in terms of Engler degrees.
2. If the viscosity of a sample of liquid petrolatum is found to bc less thnn
the official requirement, what does the determination indicate relative to
the composition of the oil?
3. Would viscosity requirements be of value 'as a constant for (1) glycerin,
\2) castoI' oil, (3) simple syrup?
 CHAPTER XVII
PHOTOMETRIC METHODS OF ANALYSIS
Photometric methods of chemical analysis are based upon the
meaSl1l'emcnt of the changes in the amount or character of light
caused by chemical reactions. Changes in the amount of light
may be due to absorption or reilection. Measurements based
on the absorption of light are termed colorimetric methods, and
the process is knowll as colorimetry. Measl1l'ements based on
the reflection of light are termed nephelometric methods, and the
process is termed nephelomct7y.

FIG. 37.-Nessler apparatus.

Colorimetry.-Thc usc of color as a moallS of determining the

amount of a given constituent in a substance has been utilized
for many years in t=malytical work. It is only in recent times,
however, that the methods have been extended and refined.
Colorimetric methods of analysis generally consist in adding
a reagent to a solution of the substance being tested so that a
color is produced and comparing the intensity of color in the
product with that produced by adding the same reagent to a
solution of the substance of known concentration. Frequently,
they are rapid and accurate; however, their greatest advantage
is their extreme sensitivity which makes it possible to determine
very small amounts of substance, in many cases unattainable by
other methods.
260
 PIlOTOME1'RIG' METHODS OF' ANALYSIS 261

The simplest type of apparatus for colorimetric measurements

consists of the Nessler tubes and rack (Fig. 37). The tubes have
parallel sides and flat bottoms and are graduated to 50 and 100
ce. Nessler tubes ilhould be of the same diameter, but frequently
they are not. Consequently, they should be compared for
equality of distance between the 50 ancilOO cc. graduation marks
and only those which agree should be selected. The use of this
apparatus is illustrated in the following exercise.
Exercise 81
Object.-To Determine the Ammonia Content of Water.
Materials Required.-Natural water (use tap water).
Ammonium ehloride solution. Dissolve 0.3141 Gm. of pure ammonium
chloride in ammonia-free water and dilute to 100 ce. Dilute 10 ce. of this
solution to 1,000 ce. with ammonia-free water. Each cubic centimeter of
the resulting solution contains the equivalent of 0.01 mg. of NH a
Rochelle salt solution. Dissolve 5 Gm. of potaesiulll and sodium tartrate
in 10 cc. of water.
Nessler's reagent. Dissolve 2.5 Gm. of potassium iodide in 3 ce. of
water, add 3.5 Gm. of mercuric chloride, and sti~ until solution is complete.
Add 100 cc. of a 25 per cent solution of potassium hydroxide, mix, Itilow to
settle, and decant the clear supernatant liquid. Preserve the Holution in 8
rlark place.
Ammonia-free water. Test the distilled water in the laboratory by
placing 50 cc. in a Nessler tube previously washed several times with th~
distilled water and add 2 ce. of Nesiller's reagent. Cover the Lube antI
allow it to stand for 5 min. If the color produced after standing is mor(1
intense than that of the Nessler's reagent, immedilltely after dilution, thl1
water must be purified. If purification is necessary, add 1 Gm. of
potassium permanganatc to 1,000 cc. of tho distilled water, di~'til and whell
the distillate does not give a test for ammonia wit,h Nessler's reagent, Lallcet
the water in a clean bottle previously rinsed with a portion of th' 'lnnnonia-
free distillate.
12 Nessler tubes.
A Nessler tube stand.
Procedure.-Place 50 cc. of the water to be tested in one Nessler tube
Place 0.1, 0.2, 0.3, 0.4, 0.5, etc., cc. of the ammonium chloride solution ill the
other eleven Nessler tubes and add ammonia-free distilled wat.er up to the
50 cc. mark. Add 2 ca, of Rochelle salt solution and 2 cc. of Nessler'~
reagent to each of the tubes. Mix the contents of the tubes thoroughly and
place them in the stand. While looking vertically down through the solu-
tion, vary the standard color solutions until the color of the unknown
matches one' or lies between two of them. Calculate the milligrams pel'
liter of ammonia in the sample of water tested.
::l62 ()l!flN'l'I1'A1'IV]!] PlL1RMJ1(!EU'l'ICAL CHEMIS'l'RY

When comparing the intensity of color in the tubes, place them

jn the stand and adjust the position of the latter in front of
a window so that light from the opaque reflector will pass upward
through the tubes.
Calcium and magnesium are precipitated from aqueous solu-
tion by Nessler's reagent. The Rochelle salt is added to prevent
this.
Nessler's solution reacts with the ammonia and ammonium
salts to form an orange-colored compound of the composition
HgO.Hg(NH2)I. When Nessler's reagent is added to solutions
containing more than a few
milligrams of ammonia per 100
ce., a difltinct orange-colored
precipitate is formed, and very
small concentrations of ammo-
nia, such as 1 part in 1,000,000,
produce a distinct yellow to
brown coloration.
The number of milligrams
of ammonia per liter of water
may be calculated as follows:
Assume that the color produced
in the sample exactly matches
that in the tube containing 0.5
cc. of the ammonium chloride
solution. Since each cubic
centimeter of ammonium chlo-
ride solution contains the equiv-
Fw. 38.-Duboscq colorimeter. alent of 0.01 mg. of NH3 per
cubic centimeter, 0.5 ce. will
contain the equivalent of 0.005 mg. per cubic centimeter or 5.0
mg. per liter. The sample of watel' must contain the same
amount or 5 mg. per 1,000 cc. equivalent to 5 parts of ammonia
in 1,000,000 parts of water. .

Questions and Problems

1. Whit is meant by the term colO1'irnetry?
2. Look up" Aqua" in the U.S.P. and explain the use of Nessler's reagent.
in the tests for purity.
 PHOTOMETRIC METHODS OF ANALYSIS 263

3. Look up Beer's law in a textbook of physies and explain what applica-

tion if any is made of that law in colorimetric analysis.
4. Review the Gunning-Kjeldahl method of estimating nitrogen in organic
compounds and briefly explain how that method combined with thc eolori-
metric method might be used to determine the nitrogcll content in very
small amounts of certain organic compounds.
Colorimeters.-A number of colorimeters have been developed
for rapid and precise colorimetric measurements. The Duboscq
colorimeter (Figs. 38 and 39) is n,
device of :this type, and since it is
generally used, its construction and
use will be described here. The
essential parts of the instrument
consist of plungers, cups, a reflecting
m i 1'1' 0 I' , a compound prism, an
observation microscope, and adjust-
ments. Two prisms of optical glass,
hexagonal in form, matched for color
and with optically plane and parallel
ends are attaehed to the frame of
the instrument by threaded metal
adapters. Two detachable cups in
which the liquids under examination
are placed are threaded on movable
stages beneath the plungers. The~e
cups are made of glass fitted into "===~R=1B'
metal casings and have bottoms ~1llI
made of optically inactive glass
disks. An adjustable mirror with
two reflecting surfaces is so placed
that light can be reflected through
the cups. A compound prism is
built in above the plungers in a
dust-proof housing in such a manner FIG. 39.-Diagram of the Du-
bo~cq colorimeter showing the
as to bring the light beams through path of light..
each cup to a common axis, light
from one cup illuminating one-half of a circular field and light
from the other cup illuminating the other half. An observation
microscope, by which the observeI' views both fields of illumi-
nation with one eye, is focused on the two fields. Adjustments
264 QU.1NTIT11TIYE PHARMACIWTICAL CHEMISTRY

are provided whereby the cups may be raised or lowered, the

kvel of each cup being shown on a scale and vernier.
Exercise 82
Object.-To Determine the Amount of Epinephrine. Hydro-
chloride in Solution of Epinephrine Hydrochloride.
Materials Required.-0.050 Gm. of pure epinephrine.
0.1 N hydrochloric acid.
10 ec. of 1 pel' cent solution of potassium iodate.
Commercial epinephrine hydrochloride solution 1. to 1,000.
A Duboocq colorimeter.
Procedure.-l, Place 0.050 Gm. of pure epinephrine in a 50 cc. vohimetric
Hask, add 0.5 ec. of 0.1 N HCl, and dilute the solution with sufficient dis-
tilled water to make 50 cc.
This standard solution of epinephrine hydrochloride containing
1 part of epinephrine to 1,000 parts of solution must be prepared
freshly as needed.
2. Place 20 cc. of disWlecl water in a 50 ec. Erlenmeyer Hask, add 5 ee.
of 1. per cent pot[1~Hium iodate solution and 0.25 cc. of 0.1 N hydrochloric
acid, and warm the solution to 38C. Then add exactly 0.5 ce. of the
standard epinephrine solution and heat the resulting solution at about
38C. for exactly 15 min. Prepare another acid solution of potassium iodate
in the smIle manner, warm it to 38C., add exactly 0.5 ce. of the sample of
epinephrine hydrochloride to be tested, and hetLt this solution at 38e. for
exactly 15 min.

The greatest intensity of color is produced after the epinephrine I

has been heated for about 10 min. at 38C. It is important

that the unknown and the standard solution be heated at the
same temperature for the same length of time.
3. Adjust the colorimeter, phwe it before an ample source of light, pref-
erably diffused daylight, and regulate the mirror so that the two halves
of the field are of the same color ancl equal brightness. Fill one of the cups
about one-half fullwHh the known solution and the other cup about;o~e
half full of the unknown. Replace the cups in the instrument and raise
them .until the plungers come in contact with the bases. Adjust the liquid
depth in the cup containing the known solution to any convenielit level,
preferably to an exact number of millimeters.' Raise or lower the cup
containing the unknown liquid until both halves of the field are of the same
color and intensity. Take three independent readings of the instrument
with the cups set at different levels. Calculate the percentage of epinephrine
in the unknown solution.
 Pl101'OME1'RlC ME1'lloD8 Oli ANALYSIS 265

In adjusting the colorimeter, make certain that the plungers

and cups are fitted tightly. Raise the cups ulltil the plungers
come in contact with the cup bases. The verniers 8hould then
read zero. The best source of illumination is diffused daylight
obtained by placing the polorimeter before a north window,
but that produced by an electric colorimeter lamp may be used.
The colorimeter cups should never be filled much more than
one-half full, otherwise their liquid contents may overflow when
the cups are raised. After placing the known and unknown
liquids in the colorimeter cups, raise the latter until the plungers
come in contact with the cup bases to force any bubbles of air
out from beneath the ends of the plungers.
In taking readings, set the level of the cup containing the known
liquid at some definite position, e.g., 15 111m., and regulate the
level of the cuP. containing the unknown liquid until a color
match is secured. Repeat this procedure with the cup containing
the known liquid set at 20 and 25 mm. readings. When the two
halves of the field of the colorimeter are of the same color and
intensity, the color intensities of the two solutions are inversely
proportional to the concentration. If C1 and D1 are the respec-
tive concentration and depth of the known solution, and C2 and
D2 are the corresponding symbols for the unknown, then C 1 X
D1 = C2 X D 2 If C1 = 1,000 mg. per liter, D1 = 20 rom., and
D2 = 25 mm., then: 1,000 X 20 = 25 X C2, and

C2 = 1,00025X 20 = 800 mg. per lt

1 er,

the concentration of the unknown.

Questions and Problems
'1. 'Is the principle involved in colorimetric measurements with the
Duboscq colorimeter the same as when Nessler tubes are used?
2. What advantages and disl1dvRntages does the eolorimetl'ic method of
analysis of epinephrine have in comparison with the biological method?
3. Consult the reference at the end of this chapter, and give several
!!:pplications of colorimetric methods of analysis to pharmaceutical produets.

Exercise 83
Object.-Assay of Crocus for Color.
Materials Required.-O.l Gm. crocus in fine powder.
266 QUA.NTITA.TIVE PHARMACEUTICAL CHEMISTRY

10 cc. of 0.01 N potassium dichromate.

A Nessler apparatus or a colorimeter.
Procedure.-" Weigh accurately 0.1 Gm. of Crocus, in moderately fine
powder, and macerate it at room temperature in 100 cc. of distilled water for
three hour:,; with frequent shaking. Immediately filter the mixture and add
sufficient distilled water through the filter to make 100 co. Dilute 10 cc. of
this filtratc, aceurately measured, to 100 cc. with distilled water. Immedi-
ately compare the color of this solution, in Nessler tubes or in a colorimeter,
with the (Jolo!' of hundrodth-normal potassium dichromate.
"The color tint of the dilute Crocus solution approximates that of the
hundredth-normal potassium dichromate; the strengt,h of col01', when
expressed in illlll. of column height, shall be not less than that of the hun-
dredth-nol'llHll plltassium dichromate."

See Exercise 81 (page 261) for directions and explanation of the

use of the Nessler apparatus and Exercise 82 (page 264) for
direction~ and explanations of the llHe of the colorimeter.
Cudbear and tincture of cudbear of the N.F. are also assayed
for color by similar procedures. Other simple color comparisons
are used in the official tests for carbonizable substances. These
qualitative tests are based upon matching the color developed
when a substance is treated with sulfuric acid with that of various
standard matching fluids. (See U.S.P. XI, page 557.)
Questions and Problems
1. What are colorimetric solutions? (See U.S.P.)
2. Look up the official tests for limit of lead and carbonizable impurities in
the U.S.P. Are these colorimetric tests?

Nephelometry.-Nephelometric measurements afford a means

of determining directly the mass or weight of a precipitate in
l3uspension, i.e., without filtering, drying, and weighing. The
method commonly used is based upon the measurement of the
brightnesl3 of the light reflected by a cloud of finely divided
particles suspended in a liquid. When other conditions remain
constant, the intensity of the reflected light is a function of the
amount of suspended particles.
Nephelometric methods of analysis have been applied to a
great number of inorganic and organic compounds, such as
chlorides, sulphates, ammonia, phosphorus, silver, proteins, fats,
volatile oils, and alkaloids. Many substances can be deter~
mined quantitatively by nephelometric methods in concentrations
of 0.1 to 1 part or less in 1,000,000 parts of water; e.g., 1 part of
 PHOTOMETRIC METHODS OF ANALYSIS 267

phosphorus in 330,000,000 parts of water has been determined

by this method. Generally nephelometric methods are limited
to measurements of suspensions of substances in low concentra-
tion, usually not greater than 100 mg. per liter. Larger amounts
of substanccs can be estimated, however, if properly diluted.
The instrument used for nephelometric measurements may
consist of a Duboscq colorimeter fitted with special nephelom-
eter attachments. These attachments include two cups with
transparent sides and black bottoms in place of the colorimeter
cups; two tubular metal light shields which surround the cups in
such a way that the depth of the liquids illuminated by a light
source may be varied; a special nephelometer illuminator; two
condensers by means of which a pair of parallel rays are projected
horizontally into the cups containing the liquids under examina-
tion. Nephelometric determinations preferably should be carried
out in a dark room.

Exercise 84

Object.-To Determine the AmOlmt of Arsenic Trioxide in

Solution of Arsenous Acid.
Materials Required.-25 ce. of potassium molybdate solution, 1 pel' cent.
50 cc. of 1.0 N hydrochloric acid.
25 cc. of cocaine hydrochloride solution, 2 per cont.
0.1 Gm. of pure arsenous acid.
10 cc. of solution of arsenous acid.
A Duboscq nephelometer with illuminator aUacllluents.
Procedure.-1. Place 25 cc. of 1 per cent potassium molybdate solution
in a 100 cc. flask, add 50 cc. of 1.0 N hydrochloric acid and 25 cc. of 2 per
cent cocaine hydrochloride solution. Shake the mixture thoroughly and
preserve it in a tightly stoppered bottle in a dark place.

This mixture constitutes th~ nephelometric reagent for arsenic.

It will keep for a long time if properly preserved. Inunediately
prior to using a portion of this solution, it should be filtered from
any sediment present, using a quantitative filter paper.
2. Dissolve exactly 0.1 Gm. of pure arsenic trioxide, previously dried to
constant weight at 100C., in sufficient distilled water to make 100 cc.

This is a standard arsenic trioxide stock solution containing

1 mg. of As 2 0 s in each cubic centimeter. Other standard
268 QUl1NTl7'11'l'IVE PHl1RMACEU'1']CAL .CHEMISTRY

solutions contai:ning 0.1,0.01,0.001 mg. of As 20 a can be prepared

by dilution in the proper ratio.
3. Dilute 10 ceo of the standard arsenic trioxide solution to 1,000 cC. with
distilled wntel'. Place 10 cC. of this diluted solution in a 30 cc. glass-
stoppered bottle, add 2.5 cC. of the c:ocaine-molybdeIllun reagent, and shake
the mixture vigorously. Dilute 1.0 ce. of the unknown arsenous acid solu-
tion to 1,000 cc. ltnd treat a 10 cc. portion of this solution with the cocaine-
molybdenum reagent ill the same manner and at the same time as the
~tandard.

The diluted standard arsenic trioxide solution should contain

0.001 mg. of As 20 a per cubic centimeter and the amount of
arRenic trioxide in the solution of arsenous acid should contain
approximately the same amount of As 20 a per cubic centimeter
if the solution is of about the official strength.
The cocaine-,molybdenum rcagent produces a cloud when added
to solutions containing arsenic trioxide. The best clouds are
obtained when from 0.05 to 0.005 mg. of arsenic are present in
each 10 cC. of the solution.
4. Pdur the known and unknown solutions of arsenic trioxide into the
respective cups, filling them to about 6 mm. from the top. Place the
tubular light shields about the cups on the cup stltge and raise them up
until the senle reading is zero. Place the instrument ill position with respeet
to the nephelometer illuminator, set the position of the cup containing the
known solution at 15 mm., and adjust the level of the cup containing the
ullimown until a balanced field is obtained. Take thrce separate mel1Stlre-
ments with the cup containing the stalldal'd solution set at 15, 20, and
25 mill.

The calculations are based on the principle that the turbidities

are inversely proportional to the length of the column of liquid.
Thus, if (JJ is the Goncentration of the standard solution, (J2
the concelltl'l.Ltion of the unknown, L1 and L2 the lengths of the
columns of liquids in the cups containing the known and unknown
solutions, respectively, the concentration of the unknown solu-
tion may be found by substituting in the following formula:
O2 =0 0 1 XLI
L2
Questions and Problems
1. Would it be practicable to repll1ee the present offioial test for arsl)nic
(see U.S.P., page 436) by a nephelometric method?
 PHOTOMETRIC ME1'IIODS OF ANALYSIS 269

2. Name saventl official, nl'senic-contllining preparations to which this

method with slight modifications might he applied.

Exercise 86
Object.-To Determine the Amount of Oil of Peppprmint in
Spirit of Peppermint.
Materials Required.-5 ce. of spirit of peppermint.
1 cc. of oil of peppermint.
Diluted hydrochloric acid.
Alcohol, 95 per cent (redistilled).
Procedure.-Dissolve 1 ee. of oil of peppermint, accllmtely mea~un'll
from a pipette, in sufIicient 95 per cent alcohol to make exactly 10 (~t:.
Dilute 5 co. of this solution with 25 cc. of diluted hydrochlorie acid, ehftke
thoroughly, and compare by mef1IlS of a nephelometer the turbidity pro-
duced with thnt of an equctl volume of spirit of peppermint dilutccl in the
slLlne 111anner.

When the alcoholic solution of volatile oil is diluted with

hydrochloric acid, the volatile oil is thrown out of solution in
the form of finely divided globules with the formation of a fairly
stable emulsion. The amount of oil present is then estimated by
measuring the amount of light reflected from a column of the
emulsion by matching the latter with the light reflected from a
column of emulsion containing a known amount of oil. The
measurements should be made as soon as possible after throwing
the oil out of solution so that there will be no error caused by
breaking of the emulsion.
Turbidimetric Tests.-Turbidimetric tests are applied to
certain official chemicals to insure the absence of excessive
amounts of ehloridB and sulfate. The following directions are
given in the U.S.P. for the performance of these tests:
"In carrying out these turbidimetric tests, the following points
are to be observed: The same quantities of the same reagents
must be used in the test of the substance under examination and
in the control test. The glass cylinders in which the tests
are made must be of the same diameter and mateh in all other
respects as closely as possible. The precipitating reagent must
be added to both in immediate sequence.
"Chloride.-The prescribed quantity of the substance to be
tested is dissolved in from 30 to 40 ec. of distilled water, I1nd the
solution neutralized, if necessary, with nitric acid, using litmus
270 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

paper as the indicator. One cubic centimeter of nitric acid and

1 cc. of silver nitrate T.S. are added and then sufficient distilled
water to make 50 cc. After mixing well and allowing to stand
five minutes protected from direct sunlight, the turbidity, if
any, is compared with that produced in a control test made with
the specified volume of fiftieth~normal hydrochloric acid.
" Bismuth Salts are first dissolved in a few cubic centimeters of
distilled water and 2 cc. of nitric acid, then diluted with distilled
water to 50 cc.
"Sulfate.-The specified quantity of the substance to be tested
is dissolved in from 30 to 40 cc. of distilled water, and the solution
neutralized, if necessary, with hydrochloric acid, using litmus
pf~per as the indicator. One cubic centimeter of diluted hydro-
chloric aeid and 1 cc. of barium chloride T .S. are added and then
sufficient distilled water to make 50 ec. After mixing well,
it is allowed to stand for ten minutes and the turbidity, if any,
is compared with that produced in a contresl test made with the
specified volume of fiftieth~normal sulfuric acid.
" If the solution, after the addition of acid, is not perfectly
clear, it is filtered through a filter paper free from chloride or
sulfate, then the silver nitrate or the barium chloride is added.
"In applying the turbidimetric tests to salts of heavy metals,
which normally show an acid reaction, their aqueous solutions
prepared for the test are not to be neutralized.
"When the tests for chloride or sulfate are to be applied to a
specified volume of a solution of a substance prepared as directed
in the text, and the permissible limit for these impurities cor-
responds to 0.2 cc. or less of fiftieth-normal hydrochloric or
sulfuric acid, the solution is not to be further diluted. The con-
trol test is also made with the same volume of water (or other
specified solvent) as in the test."
Exercise 86
Object.-Limit Test for Chloride and Sulfate III Calcium
Gluconate.
Materials Required.-3 Gm. of calcium gluconate.
0.02 N hydrochloric acid .
. 0.02 N sulfuric acid.
Silver nitrate 'r.S. 1:1. N silver nitrate).
 PHOTOMETRIC METHODS OF ANA.LYSIS 27!
Barium chloride T.S. (12 Gm. of barium chloride in suffieient water to
make 100 ce.)
Nessler apparatus.
Procedure.-Proceed as directed above using 1 Gm. of calcium glnconate
and 1 cc. of 0.02 N HCI in the test for chloride and 2 GIll. of calcium glucon-
ate and 1 cc. of 0.02 N H 2S0 4 in the test for sulfate.

Questions and Problems

1. Calculate the pel' cent of chloride and of sulfate permitted in calcium
gluconate in the official test for purity.
2. List several official substances which are required to show no turbidity
when tested for chlorides and sulfates.
3. How could the volatile oil content of a highly colored solution he
determined nephelometrically?
4. The presence of sugar in a solution contl1ining volatile oils alters the
particle size when the oil is thrown out of solutioIl. How could the volatile
oil content of a cordial be determined by means of the nephelometric method?
5. Acetone is precipitated from solutions containing as little as 1 part in
100,000,000 by silver-mercury cyanide solution. Could the nephelometric
method be used to estimate the presence of small amounts of acetone in
alcohol, in ether, and in urine?
6. How might minute quantities of each of the following ions be deter-
mined: iodide, sulfate, phosphate, and calcium?
7. Could minute quantities of alkaloids in solution be estimated nephelo~
IlJ.etrically? Suggest possible precipitants.

Those who desire to make a more extended study of photomet-

ric methods of analysis will find a very comprehensive treatme:nt
of. the subject and an extensive bibliography in "Photometric
Chemical Analysis," by J. H. Yoe (2 vols., John Wiley & Sons,
Inc., New York, 1929).
 CHAPTER XVIII

DETERMINATION OF HYDROGEN ION CONCENTRATION

Eleetromctric methods of analysis may be divided into two
classes, namely: (1) conductimetric methods in which the end
point is detected by measuring the change in electrical con-
ductance during a titration; (2) potentiometric methods in which
the end point is found by measuring the change in potential of a
suitable electrode during the titration. The latter has become
of increasingly great importance in all fields of chemistry within
comparatively recent years.
Portions of the theoretical considerations ()f pH which
follow have been taken with slight changes from the U.S.
Pharmacopoeia.
Many of the compounds which are pharmaceutically important
are acids, bases, or salt8 thereof. In aqueous or hydroalcoholi{J
solution they tend, in great or small degree, to dissociate into
their respective ions. The concentration and activity of hydro-
gen ions in a solution influence the concentrations of the anions,
cations, and undissociated molecules present in :the solution.
These factors, in many instances, affect the stability, therapeutic
activity, and pharmaceutical elegance of medicaments in aqueous
or hydro alcoholic solutions.
Acid~Base Equilibrium and pH.-The dissociation of acids
may be represented by the type description:
AH >H+ -I- A-

In a similar manner the dissociation of substituted ammoniums

may be represented:
BHI )B -I- H+

The dissociation of the metallic hydroxides may be represented:

BOH )B+ -I- OH-

272
DETERMINATION OF HYDROGEN JON CONCEN'l'RATlON 273

For the purpose of this discussion, salts, certain acids, and the
hydroxides of -the alkali metals are aflsUIned to be completely
dissociated in dilute aqueous solution.
At equilibrium the orclinl1ry mass aetion equation will hold us a
first approximation for weak acids
[H+] [A -] = Ka
(1)
[HAJ
[A-] Ka
[HA] = [H+]' (2)

where the brackets indicate molar concentration of each ionic or

molecular species.
The constant, IC, is the dissociation constant of the aeid. The
value of Ka is markedly different for various acids, and for the
same acid it is appreciably affected by the presence and concen-
trations of various ions in solution. The ratio [A-l/[HA1 in
equation (2) is seen to depend upon the value of IL and the con-
centration of hydrogen ions. Assuming that Ka remains con-
stant, the ratio [A-]/[HA] can be changed only by changing the
value of [H+]. This can be accomplished by the addition of a
strongly dissociated acid which increases [H+] or the addition of a
strongly dissociated base which will decrease the value of [H+].
Furthermore, the ratio [A-]/[HA] may be changed by the addi-
tion of the acid itself or its salt, but not directly as the ratio of
these added quantities, for [H+] will have been changed also.
The magnitudes of [H+] vary enormously, i.e., from one to one
ten-millionth squared or 10- 14 , The desirability of an exponen-
tial or logarithmic seale of expression is at once apparent. The
system generally adopted is that devised by Srenson, namely,
the use of the symbol pH, which is defined as the negative logarithm
of the hydrogen ion concentration (see page (7). Hence:
1
pH = -log [H+] 01' log [H+] (3)

The relation of pH to [H+] may be seen in Table XXXVII.

The logarithmic nature of this system should be borne in mind
constantly ill considering changes in hydrogen ion concentration
. and its connotation in terms of pH as illustrated in Table
. XXXVIII.
274 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

TABLE XXXVII

Normality, Normality,
pH hydroxyl ions
hydrogen ions

0 1 lO- u
1 10-1 10-13
2 10- 2 10-12
3 10-3 10-11
4 10- 4 10-10
5 10-5 10-9
6 10- 0 10-8
Neutral point 7 10-7 10-'/
8 10-8 10- 0
9 .10- 9 10-5
10 10- 10 10-4
11 10-11 10-3
12 10- 12 10-2
13 10-13 10-1
14 10-14 1

TABLE XXXVIII
[H+j pH
Times Increased or Decreased Corresponding Change
10 1.00
5 0.70
2 0.30
1.5 0.18
1.1 0.04
1. 05 0.02
1.023 0.01

Equation (2) may be written

[A-] 1
log [HA] + log Ka. = pH (4)

Obviously when the ratio [A-]/[HA] is unity, log I~a will be

equal to pH. Also, when pH is greater than log ia by two units,

practically the entire compound exists as species A-. When pH,
however, is less than log i"
by two units, pl'actically the entire
compound exists as species HA. When pH has a value lying
DETERMINATION OF HYDROGEN ION CONCENTRATION 275

within 2 units log I~a' there will bc relatively large proportions

of each species present.
The standard device generally used in the determination of
hydrogen ion concentration is the hydrogen half-cell.
A hydrogen half-cell consists of a noble metal electrode,
immersed in a solution containing hydrogen ions or substances
capable of supplying such ions at the activity symbolized by
(H+), under a definite partial pressure of hydrogen, PH2, and sup-
plied with a catalyst such as platinum black to facilitate the
half-reaction
H2 2H+ + 2e
When two such half-cells are placed in liquid junction, the
electromotive force E of the cell is defined by

E = RT In (H+)'~ +E (5)
F (H+h/p'H2 L,

Where (H+), is the activity of the hydrogen ions in the half-

cell having the greater hydrogen ion activity, P'H2 is the hydrogen
pressure in the same half-cell and EL is the potential jump at the
liquid junction of the half-cells. The latter is not eliminated
but is reduced to a small, usually neglected value, by making the
junction with saturated potassium chloride solution.
The ultimate standard of reference is the hydrogen half-cell
in which the hydrogen ion activity is unity and the hydrogen
pressme is one atmosphere. If this half-cell were used in con-
junction with another half-cell of lower hydrogen ion activity and
the hydrogen pressure were the same in the two half-cells, then
RT 1
E = yIn (H+) or (6)

1
E = 0.0581 log (H+) (7)

The foregoing ultimate standard of reference is not used in

practice and it is impossible either to eliminate entirely the liquid
junction potential or to calculate it with exactitude. Therefore
276 QUANTITATIVE PHARMACEU'l'lC'AL CHEMISTRY

for official purposes the following is adopted for approximate

measurements.

Hg HgoCb, r
KOl (sat. soln.) I Solution
k Ilown of I
-
I Ull p H Pt, H2 (1 atmos.)
Employing this cell at 200., pH may be calculated by the
following equation:
E - 0.2488
pH (8)
0.0581
The electromotive force is measured by means of a potentiometer.
When the standard 0.05 M potaHsillm biphthalate solution
(page 296) is employed in the foregoing cell, in place of the solu-
tion of unknown pH at 200., the cell has an electromotive force
of 0.4797 volt, corresponding to pH 3.974.
These half-cells employed under the prescribed conditions con-
stitute the reference device for pH measurements for the official
solutions. For official purposeR, various electrodes may bp
employed at the discretion of the operator which are capable of
as great a degree, or a greater degree, of accuracy than that
set forth in the colorimetric method, namely, 0.1 pH.
When the electrometric method is employed for hydroalcoholic
solutions and the electromotive force developed converted into
pH, the cOllversion is based on the assumption that the equation
holds for hydroalcoholic solutions as it does for aqueous solutions.
Stl'ictly speaking this condition does not obtain. Therefore
the values expressed as pH are relative and subject to change with
val'ying concentrations of alcohol.
The term pH is expressed generally by an integer and two
significant decimals. The reliability of the value of the figures
in the second decimal place is dependent largely on the method
employed in carrying out the determination. For official pur-
poses, the expression is given with one decimal only. For prac-
tically all purposes, this degree of accuracy is considered sufficient.
However, the method described is capable of a higher degree of
accuracy.
The potentiometric method of analysis may be applied to
determine the end point of titrations and to determine the
hydrogen ion concentration of solutions. The hydrogen iOll
DE1'ERMINA'l'ION OF HYDROGEN ION CONCEN1'RA1'lON 277

concentration of solutions can also be determined eolorimetri-

cally (see page 297). The potentiomctric method, however, is
more accurate in general and is. to be preferred even though it
rcquires a more elaborate apparatus and more experience on the
part of the analyst.
Potentiometric Methods.-When an acid or an alkali is dis-
solved in water, the characteristic properties of the solution
obtained are due to the hydrogen and hydroxyl ions formcd by
the dissociation of the acid or base. The effec.tive acidity
or alkalinity depcnds on the extent of dissociation of the acid
or bas_e in solution and is independent of the total ~Cicljty or
alkalinity. In the volumetric estimations of acids and alkalies,
the total acidity or alkalinity is determined. In measuring
the effective acidity or alkalinity (the strength of the aeicl or
base) of a solution, however, it is necessary to determine the
hydrogen ion concentration.
The hydrogen ion concentration of an aqueous solution may be
determineu by measuring the voltage developed between two
electrodes of special character immersed in the solutioil by means
of a potentiometer, since the voltage developed between the
electrodes directly depends on the concentration of hydrogen
ions in the solution. The measurement of the voltage produced
between the electrodes is equivalent, therefore, to a determination
of the hydrogen ion concentration and to the determination of the
effective acidity or alkalinity of the solution.
The electrodes employed in the determination of hydrogen ion
concentration must be of such character that one of them will
develop a potential that varies according to the concentration of
hydrogen iOlW, and the other must have a constant potential
unaffected by the hydrogen ion concentration. Although a num-
ber of electrodes of different character have been developed for.
use in potentiometric methods of hydrogen ion concentration
measurements, the fundamental method involves the use of the
so-called hydrogen electrode and the saturated calomel electrode
described here.
The hydrogen electrode (Fig. 40) consists of a small square of
platinum foil attached to a short piece of platinum wire sealed
into and extending through one end of a glass tube. Mercury
is poured into the open end of this tube to cover the platinum wire
278 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

in the bottom so that the electrode may be connected with

the potentiometer by dipping a wire in the mercury. This
electrode element is placed in another tube of larger diameter
through a side tube of which purified hydrogen gas is conducted
to the electrode when it is immersed in a solution.
Holes in the side of this outer tube at the level of
the electrode element permit the hydrogen gas to
escape witbout forcing the solution completely out
of contact with the platinum.
When the platinum tip of this electrode is coated
with platinum black (platinum in a finely divided
state), the platinum black absorbs and holds a
relatively large volume of the gas. Consequently,
when an electrode prepared in this manner is
saturated with hydrogen gas and immersed in a
FIG. 40.- solution containing hydrogen ions, it behaves as a
H y d r 0 g e n hydrogen electrode and develops a potential that
electrode.
is definitely related to the hydrogen ion concen-
tration of the solution.
To platinize the hydrogen electrode, clean a new electrode by
dipping it in hot solution of potassium dichromate, then in
concentrated sulfuric acid, and finally wash it thoroughly with
distilled water. This procedure removes any organic matter
adhering to the platinum. The platinum black can be removed
from old electrodes by electrolysis in concentrated hydrochloric
acid, the electrode being used as the anode. In the latter case,
the electrode should be removed from the hydrochloric acid and
washed with water as soon as all of the platinum black has
dissolved; otherwise the platinum of the electrode will go into
solution.. Dissolve 3. Gm. of platinic chloride and 0.010 Gm. of
lead acetate in sufficient distilled water to make 100 cc. Immerse
the electrode in this solution and connect it as the cathode to two
dry cells connected in series (the positive, center pole, of one cell
connected to the negative, outside pole, of the other) and use a
clean platinum wire as the anode to complete the circuit in the
Rolution. Allow the current to pass for from 3 to 5 min. or until a
thin, even deposit of platinum black covers all of the electrode
so that the sheen of metallic platinum cannot be observed.
When platinization is completed, wash the electrode several times
DETERMINATION OF HYDROGEN ION CONCENTRATION 279

with distilled water, immerse it in dilute sulfuric acid, and elec-

trolyze as cathode for 1 01'.2 min. to saturate the platinum black
with hydrogen. Suspend the electrode in distilled water until it
is used.
The calomel electrode (Fig. 41) is prepared as follows: Add 200
Gm. of pure potassium chloride to 500 ce. of hot distilled water
contained in a 1,000 cc. beaker, add 0.5 Gm. of pure mercurous
chloride (calomel) to the hot solution, and stir the mixture well.
Allow the mixture to cool to room temperature and filter out the
excess potassium chloride and calomel. Store the solution in a
well-stoppered bottle for future use. Clean the electrode
vessel thoroughly by washing it successively with cleaning
mixture and distilled water. Dry the vessel. Pour pure mer-
cury into the electrode vessel until the platinum wire in the
bottom is covered. Add a layer of mercu-
rous chloride about a quarter inch deep
over the mercury, pour some of the potas-
sium chloride-calomel solution 0 v e r it, +
stopper the mouth of the vessel, close the
stopcock, and shake the contents together,
being careful to avoid as far as possible
getting mercury and calomel into the side
arm. Allow the mixture to settle and fill
the vessel and side arm with the potassium
chloride-calomel solution, wash away any
floating calomel by overflowing, make
certain that no air bubbles are trapped FIG. 41.-Calamel eleo-
. trade.
in the side arm, and close the stopcock.
Stopper the electrode vessel until it is ready for use: Fill
the side tube connecting with the bottom of the vessel with
ordinary mercury so that the platinum wire that extends into
this tube from the vessel is covered.
Only the purest obtainable chemicals should be used to pre-
pare the calomel electrode. Specially purified mercury, calomel
produced by electrolysis, and analyzed potassium chloride can
be obtained from supply houses for this purpose.
The potential developed by the calomel electrode depends on
the concentration of potassium chloride used and is independent
of the hydrogen ion concentration of the solutions examined
2s0 ()Ui1N1'I'l'A7'lVE PllARMACEU'l'ICAL CHEMIS'l'RY

during a deternliliation. The potential also varies with the

temperature of the electrode. The following table shows the
potentials developed by the thrce commonly used calomel
electrodes at different 'working temperatures:
TABLE XXXIX
..

Potential at
Calomel electrode
200. 250.

0.1 N KCI. .................. . 0.3379 0.3376 0.3371

NICOL .... ,. ...... ,. ...... . 0.2860 0.2848 0.2835
Saturated KCI ............... . 0.2496 0.2458 0.2420

The scheme of assembly for hydrogen ion measurements is

shown in Fig. 42. A is a beaker of 50 to 200 cc. capacity con-
taining the solution being investigated. B is the hydrogen

l\!\i\llll\ml\I\\\(

'K

. 0

Cf,l~TDC;C~
OOg:,J'
"'~ 0
o
POl[NTIOMtTER.

. FIG. "42.-Scheme of assembly for hydl"ogen ion measurements.

I..

electrode. G is a cylinder of compressed, electrolytic hydrogen

equipped with a pressure-reducing valve D. The gas supply
is paB:~ed through a solution of pyrogallol in potassium hydroxide
contained in the gas-washing bottle E and then through distilled
DETERMINATION 0]1' HYDROGEN ION CONC'ENTR11TlUN 281

water contained in the srcond gas-wa~hing bottle F and finally

into the hydrogen electrode VPRsel. The hydrogen gas may be
generated from zinc and sulfuric acid, but in this case, the
hydrogen should be washed by paRsing it throug;l~ :m~cessive
gas-washing bottles containing alkaline permangallate, alkaline
pyrogallol, sulfuric acid, and water to remove arsenic, hydrogen
sulfide, etc. All of the connections in the gas train should be
tight to exclude the entrance of air. Pure gum rubber tubing,
5 mm. bore, and of medium wall should be used for the connec-
tions. H is an aspirator bottle containing a resel've supply of
saturated potassium chloride-calomel solution connected to the
calomel C'lectrode vessel so that the side arm of the latter can be
flushed out at frequent interval::; and before each new deter-
mination. The flow of solution from the aspirator bottle is
controlled by the pinchcock J.
The voltage measuring equipment consists of a potentiometer
by means of which the voltage produced by the hydrogen and
calomel electrodes dipping into t.he solution is measured by
establishing a known voltage in opposition and exactly equal to
that of the electrodes; a storage cell, or battery, K to supply a
constant source of current; a galvanometer J which shows whell
the voltage developed at the electrodes is exactly counterbal-
anced against the voltage of the potentiometer; and a standard
cell L to establish known voltage conditions in the potentiumeter
circuit.
Exercise 87
Object.-To DE'tpl'mine the End Point of Titration of Hydl'o-
chloric Acid with Sodil).m Hydroxide Potentiometrically.
Materials Required.-A potentiometer.
A galvanometer.
A standard cell.
A storttge battery.
A hydrogen electrode.
A calomel electrode.
A cylinder of compressed electrolytic hydrogen 100 or 200 cu. ft. capacity,
equipped with a pressure-reducing valve.
Two gas-washing LottJes. -
An aspirator hottle. . .
12 to 20 ft. of well-insulated, IS-gage copper wire
3 titration beakers, 50 to 200 cc. capacity.
282 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

3 ft. of rubber tubing, 5 mm. bore, medium wall.

8 oz. of specially purified mercury.
1 oz. of calomel, specially purified.
1 lb. of potassium chloride, analyzed grade.
3 Om. of platinic chloride.
0.010 Om. of lead acetate.
50 ee. of normal sodium hydroxide solution.
100 ce. of normal acetic acid solution.
25 cc. of 0.1 N hydrochloric acid.
0.1 N sodium hydroxide solution.
A number of types of potentiometers are available and used.
The potentiometer referred to here is the Leeds Northrup and'
Company modified type K instrument. The student potenti-
ometer made by the Leeds Northrup and Company is a simpler
and less expensive apparatus.
Proc~dure.-1. Assemble the apparatus, as illustrated in the diagram
(Fig. 42). Close the switch on the potentiometer that connects the standard
cell in place of the electrodes, tap the galvanometer key gently, and note
the deflection of the galvanometer. Adjust the rheostat R until the gal-
vanometer index does not move when the key is tapped.

The voltage of the potentiometer is exaetly balanced against

that of the standard cell, the voltage of which is known, when
the galvanometer index does not move upon tapping the galva-
nometer key. Do not hold the galvanometer key down to note
the deflection of the index, but tap it firmly and quickly.
2. Dissolve 40.836 Gm. of reagent potassium biphthalate in distilled
water and 'dilute the solution to 1,000 cc. Dilute 25 cc. of this solution to
100 cc. t,o obtain 100 cc. of 0.05 M potassium biphthalate. Transfer about
25 cc. of this solution to the titration beaker and immerse the hydrogen and
calomel electrodes in the solution as indicated in the diagram (Fig. 42).
Disconnect the gas train from the hydrogen electrode, open the valve of
the pressure gage, and allow a current of hydrogen to flow for about 2 min. to
sweep all air out of the tubes and wash bottles. Connect the gas line to the
hydrogen electrode immersed in the buffer solution, and regulate the flow
of hydrogen 80 that about three bubbles of gas escape from the electrode per
second. Permit the hydrogen to flow for about 5 or 10 min. to saturate the
electrode and the solution with gaseous hydrogen. Flush a small quantity
of potassium chloride through the side arm of the calomel electrode, to
remove any test solution diffused into it, and close the stopcock on the side
arm almost completely. Change the switch on the potentiometer from the
, standard cell to the electrodes, tap the galvanometer key, and note the
deflection of the index. Adjust the dial switch and slide.wire knobs on top
DETERMINA1'ION OF HYDROGEN ION CONCENTRATION 283

of the potentiometer plate until the galvanometer index shows no deflection

upon tapping the galvanometer key.

The voltage developed by the calomel and hydrogen electrodes

in this solution at 200. is 0.4797 volt with a saturated calomel
electrode corresponding to pH = 3.974. If the apparatus
measures the required voltage in this solution, it is in proper
working order for measurements in solutions of unknown hydro-
gen ion concentration.
Do not change the rheostat adjustment while testing the
apparatus. Record the voltage from the setting of the dial
switch and slide wire.
3. Check the potentiometer voltage against the standard cell again as in
step 1, and if the galvanometer index is deflected, adjust the rheostat so
that the index does not deflect. Remove the test solution from the titration
beaker, wash the hydrogen electrode with distilled water, and flush a small
quantity of potassium chloride through the side arm of the calomel electrode.
Place 25 cc. of 0.1 N Hel in a 200 cc. titration beaker and dilute it with
about 80 cc. of distilled water. Determine the voltage developed by -the
hydrogen and calomel electrodes immersed in the acid solution. Accurately
measure about 20 cc. of 0.1 N sodium hydroxide solution into the titration
beaker and again determine the voltage. Add successively four accurately
measured 1 ec. portions of 0.1 N sodium hydroxide and determine the Voltage
after the addition of each cubic centimeter. Continue to add portions
of sodium hydroxide solution dropwise and determine the voltage after
the addition of each portion until 26 cc. have been added. Add 5 ec. more
of 0.1 N alkali and determine the voltage. Plot a curve using cubic centi-
meters of 0.1 N sodium hydroxide'solution as abscissae and voltage readings
as ordinates, and determine the end point of titration from the curve.
Calculate the hydrogen ion concentration and the pH at the end point.

If the normality of both the acid and alkali solutions used is

not known, it is advisable to run a preliminary test, using a
suitable indicator to determine the end point so that the equiva-
lE;nt point determined potentiometrically will not be overrun.
As alkali is run into the acid in the titration, it will be noticed
that the voltage changes slowly at first, more or less gradually
as the equivalent point is approached, very rapidly with respect
to very small amounts of sodium hydroxide at the equivalent
point, and then very slowly again as an excess of alkali solution
is added.
The pH value of the solution corresponding to the measured
voltage at any point in the titration may be found from a curve
284 QUAN'l'[Tlt'NVE PHARMACEV'l'1CAL C'IIEMIS'l'RY

showing the relationship (Fig. 43). ,If the voltage is measured

with a hydrogen electrode and a calomel electrode at 25C. the
pH value can be calculat,ed hom the following:
pH = 16.9 (V - 0.246) for the saturated calomel electrode
pH = 16.9 (V - 0.285) for the normal calomel electrode
pH = 16.9 (V - 0.338) for the O.1norrnal calomel electrode

o A. .~
rB _"-,c
\ \\
1\\~ Curve A'WifhSaturodea
KG( Calomel Efecfroc/e
Curve 8-Wifh Normal kG!
\~\ CGllomel Electrode
Curve C- Wifh Tenth Normal
~ l\ Calomel Electrocle

~ j\"" , '"
6
,
, '-~,

'\ ~
pH
7
f\\ f\
\ l\\ ,.
8
9
I\'f\
~\
0
I
'\ '\
'l ~\ 1\
\[\\
J
14 I\'l\
0.2 0,3 0,4 0.5 0.6 0.1 0.5 0.9 r.O r.l 1.2
VoltGlge
FIG. 43.-Gro,ph showing relation of pH and voltnge me!tsured with hydrogen
lwei calomel electrodes.

V is the measured v~ltage, 0.246, 0.285, and 0.338 arc the

reference potentials of the saturated, normal, and 0.1 N calomel
electrodes, respectively, and 16.9 is a factor which when multi-
plied by this difference converts the potential of the hydrogen
electrode which corresponds to the hydrogen ion concentration
of the solution into the corresponding pH value. The relation-
ship between pH and voltage measmcd at 250. with the hydro~
gCll and calomel electrode is given in the graph (Fig. 43). Some
typical titration curves are illustrated in Fig. 44.
DETERMINATION OF HYDROGEN ION CONOENTRr17'ION 285

Notes and Precautions.-l. A thin deposit of platinum black

on the hydrogen electrode is better than a thick deposit. The
former establishes equilibrium more rapidly and is not "poi-
soned" so easily as the latter.
2. It is advisable to test different hydtogen electrodes against
the same calomel electrode in the same solution for agreement in
o , t 3 4 S 6 1 8 9 m" ~ D M&~ n rn ~ m

'i
z~ -

3i
4~~~~~~~liijl!!I~~~~~~~~
5i~

1}6ill:- - - -

8i
9t -
10
1
IIi
12;
"f
14'
FIG. 44.-Characteristic titration curves. Vertioal scale shows pH values.
voltage indications. A difference in the agreement betv;reen two
hydrogen electrodes can also be detected by substituting one
for the calomel electrode and measuring the other against it.
The agreement in the voltage of two calomel electrodes can also
be determined in this way by substituting one for the hydrogen
electrode.
3. The 'true potential of the hydrogen electrode in a solution
is attained only when the electrode and solution are saturated
286 QUAN1'ITA1'IVE PHARMACEUTICAL CHEMISTRY

with hydrogen. Inconsistent results are frequently obtained in

hydrogen ion determinations due to the use of insufficient
hydrogen.
4. The hydrogen electrode cannot be used in solutions con-
taining strong oxidizing agents, such as ferric iron, dichromates,
nitric acid, and chlorine. It cannot be used in solutions con-
taining strong r~ducing agents, such as hydrogen sulfide or
sulfurous acid.
5. Certain organic compounds which are easily reduced
frequently prevent accurate results, e.g., unsaturated acids,
nitrobenzene, and aniline dyes.
6. The hydrogen electrode does not function properly in
solutions containing metal ions that fall below hydrogen in the
electromotive series of the metals or lead ions, since all of these
ions are reduced on the platinum electrode.
7. Sometimes unexpected a,nd erratic results occur in determi-
nations employing the hydrogen electrode. This may be due
to the so-called poisoning of the electrode which usually causes
a drift in the voltage readings when attempts are made to balance
the potentiometer. A number of substances are commonly
known to produce electrode poisoning, especially mercury, pro-
teins, oxygen, some alkaloids, formaldehyde, ammonia, and
hydrogen sulfide.
8. Hydrogen electrodes that have been poisoned can frequently
be restored to normal by washing them in nitric acid and caustic
alkali solution successively and then electrolyzing them in sulfuric
acid solution.
Questions and Problems
1. Study one of the first three references given at the end of this chapter
and explain tho principle of the. potentiometer.
2. Explain why a platinized platinum electrode saturated 'with hydrogen
gas acts as a hydrogcn electrode.
3. Consult the reccnt journal literature (during two preceding years)
and briefly summarize the articles treating of the application of hydrogen
ion concentration measurements to pharmaceutical prodncts.
4. In the potentiometric titration of 25 cc. of 0.2680 N H 2S0 4 with NaOH
solution, using the hydrogen electrode and a saturated calomel electrode,
the following are values in millivolts for the corresponding volumes of N aOH:
0.0 ce. = 370.0; 5 cc. = 378.1; 10 cc. = 388.0; 15.0 cc. = 397.6; 20 cc. =
406.0; 25.0 ce. = 420.2; 28.0 ce. = 460.1; 30.0 cc. = 516.1; 30.5 eC. =
DETERMINATION OF HYDROGEN ION CONCENTRATION 287

690.5; 31 cc. = 860.1; 35.0 cc. = 949.5; 40 cc. = 965.9; 45 cc. = 982.2;
50 cc. = 992.1. Plot the millivolts of electrocle potcntinl as ordinates and
the cubic centimeters of NaOH as abscissae. Determine the pH value at
the end point fron"t the eurve and calculate the normality of the sOllium
hydroxide solution.
Exercise 88
Object.-To Determine the End Point of Titration of Acetic
Acid with Sodium Hydroxide Potentiometrically.
Materials Required.-The same as in Exercise 87, except that the 0.1
N hydrochloric acid is replaced by a solution of acetic acid of unknown
concen tration.
Procedure.-Procecd as in Exercise 87 .

. Questions and Problems

1. Plot a curve using pH values as ordinates and cubic centimeters of
alkali as abscissae.
2. Look up the pH at which phenolphthalein and methyl orange change
eolor, mark the eorresponding positions on the curve obtained, and explain
which indicator is suitable for use in the titration of acetic aeid with sodium
hydroxide.
S. Why does the voltage increase more rapidly at the beginning 'Of the
titration in tIllS case than it docs in the titration of hydrochloric acid with
sodium hydroxide?
Exercise 89
Object.-To Determine the pH of Saturated Boric Acid Solu-
tion, and Physiological Salt Solution.
Materials Required.-25 or 50 cc. of each of the above solutions.
The titration apparatus described in Exercise 87.
Procedure.-Determine the pH of eaeh solution separately. Place the
respective solutions in the titration beaker, allow the hydrogen gas t.o flow
until the electrode has come to equilibrium with the voltage. Take three
voltage readings at 1 min. intervals. Calculate the pH of the solutioIl from
the data obtained.

Since the concentration of the hydrogen gas in the electrode is

constant, the potential developed varies with the concentration
of hydrogen ions in the solution and therefore may be used as a
measure of the concentration of the latter.
Questions and Problems
1. Name several official preparations to which the above method of
determining pH could be applied.
'288 QUANTI'l'A'1'IVE PIDiRMACEUTICAL CHEMISTRY

A 'considera.ble number of special electrodes have been

developed for use in potentiometric titrations in the place of the
hydrogen electrode. Thus a plain platinum wire used in place
of the hydrogen electrode is employed in oxida,tion-reduction
reactions. In pharmaceutical analysis, the quinhydrone elec-
trode has been used quite extensively in place of the hydrogen
electrode 'when difficultics are encountered in the use of the
latter.
The quinhydrone electrode consists of a piece of platinum wire
or foil, exactly the same as that used for the hydrogen electrode,
but the surface is not platinized or supplied with gaseous hydro-
gen, and it clips into a solution of quinhydrone contained in the
titration beaker.
Quinhydrone is formed from equimoleculai' proportions
of quinone and hydroquinone. When quillhydrone is dissolved
in aqueous solution, it dissociates, forming quinone and
hydroquinone:
Qllinhydrone quinone + hydl'oquinone
The equilibrium established between the dissociation products
may be indicated by the following equation:

Quinone + 2H + 2( ~ ) )hydroquinone

The ratio of quinone to hyclroquinone in acid solution is main-

tained constant by the presence of u~ldissociated quinhydrone
in solution. When this condition obtains, the. potential at the
electrode is a function of the hydrogen ion concentration of
the solution, In carrying out determinations, the potential
is measured against that of a calomel electrode, 'and the pH
value is calculated from the measured voltage in the same
manner as with the hydrogen electrode.
The potential of the quinhydrone electrode is higher than
that of the hydrogen electrode in the same solution. If a
saturated calomel electrode is used and the measurement is
carried out at 25C., the pH may be calculated from the equation:
0.453 - V 1 .
0.0591 = log H+ = pH where V is the measured voltage.
DETERMINATION OF HYDROGEN ION CONCENTRATION 289

Differences of 1 or 2C. in the working temperature du not

affect the results greatly. The pH may also be found from the
curve showing the relationship of pH to voltage for the qUill-
hydrone electrode (Fig. 45).
The quinhydrone functions well in acid solution but not in
alkaline solutions. It can be used in alkaline solution where the
pH doeR not exceed 8. Alkaline solutions can be titrated, how-
ever, by adding a known excess of standard acid and titrating
the excess acid with an alkali of known normality.
o
\ ._

2
1\\
4
\
pH
S
1\
'\
7
1\
'\
8

9
\
,
0
-0.5 -0.4 -0:.\ -m -0.1 0
1\
+0.1 +O.~
Vo I+~ ge
FlU. 45.-Graph showing the relation of pH to voltage measured with the
quinhydrone and calomel ele~trodes at 25 Q C.

Exercise 90
Object.-To Determine the pH of Elixir of Iron, Quinine, and
Strychnine, Elixir of Pepsin, and Tincture of Aconite by Means
of the Quinhydrone Electrode.
Materials Required.-A potentiometric t.itration apparatus, without Lhe
cylinder of hydrogen or the gas-washing bottles.
A plain platinum wire, foil, or gauze electrode.
50 to 100 mg. of quinhydrone.
50 cc. of each preparation.
Potassium biphthalate solution (see page 296).
290 QUANTITATIVE PHARMACEUTICAL CHEMISTRY ,

Procedure.-l. Assemble the apparatus as in Exercise 87 (page 281),

replacing the hydrogen electrode with a platinum cloctrode. Test the
accuracy of the appamtus against the standard biphthalate solution as
follo,";-s: Place sufficient of the hiphthabte solution in the titl'fltion beaker
to cover the platinum electrode completely all well as the arm of the calomel
electrode. Stir about 5 mg. of crystalline quinhydrone into the biphthalate
solution. After 3 or 4 min. measure the voltage with the potentiometer.

The surface of the platinum electrode must be clean and bright.

It can be cleaned by immersing it in boiling nitric acid, washing
it in ammonium hydroxide and distilled water successively.
The pH of the biphthalate solution corresponding to the voltage
measured can be found from the graph (page 289). When
the apparatus is working properly, the pH of the biphthalate
mixture should be between 3.97 and 3,98.
2, Remove the titration beaker containing the biphthalate mixture, wash
the pla,tinum electrode with distilled water, and flush a little KCI solution
through the side arm of the calomel electrode. Plaee about 50 ce. of the
preparation in the titration beaker and stir about 50 mg. of quinhydrone
into the elixir. Measure the voltage with the potentiometer.

Usually 2 or 3 min. are required to elapse after stirring in

the quinhydrone before the voltage becomes constant. The pH
value corresponding to the voltage measured can be found from
the graph (page 289).

Questions and Problems

1. Name several pharmacentical preparations of different types in which
the quinhydrone electrode can be used for measnring the
pH.
2. Explain how the end point in the titration of alkaloids
with acids might be determined potentiometrically.
The Glass Electrode.-It has been found that a
very thin membrane of special glass separating two
I. solutions gives potentials which depend directly on
FIG. 46,- tl h d' .
GlaSB elec- lC Y rogen lOn concentratIOns. The glass elec-
trode with an trode (Fig. 46) consists of a glass electrode bulb and
internal ele-
ment. an internal reference electrode. When the glass
membrane is mounted in a cell of the type, calomel
electrode, solution of known pH, glass membrane, solution of
unknown pH, calomel electrode, the voltage is a function of the
difference in hydrogen ion activity except for a small (I asymmetry
DETERMINATION OF HYDROGEN ION CONCENTRATION 291

potential" due to the glass. A sensitive electrometer or an

amplifier of the vacuum tube type is required to measure the
voltages because the resistance of the glass diaphragm is so great
that the ordinary galvanometer is inaccurate.
A glass electrode pH meter of the type illustrated in Fig. 47
is suitable for most pharmaceutical work where the hydrogen
electrode or the quinhydrone electrode cannot be used. Instru-
ments of this type are furnished with complete directions for

FIG. 47.-Glass electrode pH meter. (Courte8Y Central Scientific Company.)

their preparation, operation, and maintenance. Under ordinary

conditions, pH values between 1 and 9.6 can be determined with
an accuracy of 0.1 pH unit. Above pH 9.6, buffer solutions
having approximately the same positive ion concentration as
the unknowns being tested should be used to calibrate the instru~
ment. The glass electrode cannot be used to determine the pH
of alcoholic solutions. It can be used to determine the pH of
viscous and semisolid substances as well as colloidal and opaque
or colored solutions and solutions containing strong oxidizing or
reducing agents. Thus the 'pH of fats and oils, ointments, soaps,
emulsions, pastes, jellies, syrups, serums, infusions, cosmetics,
292 QUAN'l'ITA'l'IYE PHARMACEUTICAL CHEMISTRY

and many other pharmaceutical products can be determined by

means of the glass electrode.
Colorimetric Methods.-A number of colorimetric methods
for the approximl1te determiul1tion of hydrogen ioneonccntration
have been developed. The most g~nerally used of these involve
the matching of the color obtained when a definite quantity of
indicator is added to a definite volume of the liquid being tested
with that of one of a series of standard buffer solutions differing
by about 0.2 pH unit. The volume of the unknown and of the
standard buffer solution must be the same, both of them must
contain the same amount of indicator solution, and the colors
must be matched by viewing through the same depth or thickness
of solution. Except in a few cases, the limit of accuracy attain-
able by the colorimetric method does not exceed 0.1 pH unit.
For convenience, the colorimetric method of determining
hydrogen ion concentration has been adopted for official purposes
and is to be employed where pigments or proteins present in the
solution do not vitiate its use. In these instances, the poten-
tiometric method must be employed.
The principle of the colorimetric method depends upon the
assumption that an indiCf).tor in the same<concentration will
exhibit the same transition shade in different aqueous solutions
if the hydrogen ion concentrations of the solutions are the same.
Having then a series of. buffer solutions of de fin ito hydrogen ion
concentrations, comparisons can be made between the shades
of color produced in the buffer solutions and that produced by
the same indicator in the solution to be tested.
As previously stated, the KG of an acid is affected by the
ionic strength of the solution, likewise the KG of indicators is
also affected. This inflllences the color transition range of the
indicator. The electrOluetric method measures the activity
of the bydrogen ions (IF) and to obtain the same value by the
colorimetric method in solutions of various ionic strengths, a
correction, known as the 8alt~error' correction, must be employed.
In addition, the kinds of ions present in solution and their
precipitating influence on the indicator affect the accuracy of
the colorimetric method. For official purposes, where the
complexity of the medicaments makes a simple application of
these corrections impossible, the corrections are ignored.
DETERMINATION OF HYDROGEN ION CONCENTRATION 293

pH Indicators and Preparation of Their Solutions.-The

indicators llsed for colorimetric pH determinations are either
weakly acid or weakly basic. Most of the indicators used for
this purpose, however, such as the phthaleins and sulfonated
phthaleins, behave like weak acids.
The usual concentration of the indicator solution is 0.05 per
cent. From 0.1 to 0.2 ce. of the indicator solution is generally
used for 10 cc. of the liquid being examined.
Solutions of indicators of the basic type and of the phthaleins
are prepared by dissolving them in alcohol. In preparing solu-
tions of indicators containing an acid group, this group must
first be neutralized with sodium hydroxide. The procedure
is as follows:
One-tenth (0.100) Gm. of the indicator is triturated in an agate
mortar with tl~e volume of 0.05 N sodium hydroxide specified
in the following table, or with its equivalent of 0.02 N sodium
hydroxide. When the indicator has dissolved, the solution is
diluted with carbon-dioxide-free distilled water to make 200 ee.
(0.05 per cent).
The solutions should be kept in stoppered bottles, and pro-
tected from light.

T AlILE XL.-pH INDICATORS

Molec-
Indicator pH range ular . Color change Solvent
weight

--
Methyl yellow ..... ""., ... 2.9 to 4.0 225 Red-yellow Alcohol
Bromphenol blue ............ 3.0 to 4.6 (lO9 Yellow-blue 3.0 ce. 0.05 N NuOa
Methyl red ................. 4.2 to 6.3 269 Red-yellow 7.4 ceo 0.05 N NnOa
Bromcresol purple ........... 5,2 to 6.8 540 Yellow-purple 3.7 ce. 0.05 N NnOH
Bromthymol blue ........... 6.0 to 7.6 624 Yellow-blue 3.2 cC. 0.05 N NaOH
Phenol rcd ................. 6.8 to 8.4 354 Yellow-red 5.7 ceo 0.05 N NuOH
Thymol blu................. 8.0 to 9.6 466 Yellow-blue 4.3 ce. 0.05 N NaOH
Thymolphthalein ......... , .. 9.3 to 10.5 430 Colorless-blue Alcohol

Solutions Used in the Preparation of Buffer Solutions.-

Fifth molar hydrochloric acid and 0.2 M sodium hydroxide
(carbonate-free) are prepared and standardized according to
the directions given under Standard Solutions.
294 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

TABLE XLI.-BuFFEH MIX'rUHES OE' CLARK AND LUES

Hel-KCI Mixtures

pH 0.2 1Vf HOI, ce. 0.2 .iI1 KCI, ca. Dilute to, ce.

1.1 04.50 5.44 200

1.2 75.10 24.90 200
1.3 59.68 40.32 20n
1.4 47.40 52.60 200
1.5 37.64 62.36 200
1.6 29.90 70.06 200
1.7 23.76 76.24 200
1.8 18.86 81.14 200
1.9 14.0S 85.02 200
2.0 11.90 88.10 200
2.1 9.46 90.54 200
2.2 7.52 92.48 200

Phthalo.te-HCl Mixtures

pH 0.2 11f K11 Phthalate, cc. 0.2,1 HeI Dilute to, cc.
~-----~---------------I-----------

2.2 50 40.()O 200

2.4 50 a9.(l0 200
2.6 50 33.0n 200
2.8 50 2fl.50 200
:l.0 50 20.40 200
3.2 50 14.80 200
8.4 50 9.95 200

~:~_j
50 0.00 200
50 2.135 200
--------------
Phthalate-N"OU Mixtures

pH 1J.2 11f KH Phth"late, oe. 0.2 jl{ Nn.OH, 00. Dilute to, ce .
.
---.--~.

4.0 50 0.40 200

4.2 50 3.05 200
4.4 50 7.35 200
4.0 50 12.00 200
4.8 50 17.5U 200
n.o 50 23.65 200
5.2 50 29.75 200
5.4 50 35,2.1 200
5.6 .10 39.70 200
5.8 50 43.10 200
6.0 50 46.40 200
6',2 50 47.0D 200
DETERMINATION OF HYDROGEN ION CONCENTRATION 295

TAllLE XLI.-Bm'FER MDCTURE8 OF CLARK AND LUBH.-(Continlled)

KH,PO<-NIl.OH Mixtures
- ----
pH 0.2 J1f KH2PO., ce. 0.2 M NaOH, cc. Dilute to, ce.
_ . _._, _
5.8 50 3.66 200
6.0 50 5.64 200
6.2 50 8.55 200
6.4 50 12.60 200
6.6 50 17.74 200
6.8 50 23.60 200
7.0 50 29.54 200
7.2 50 34.90 200
7.4 50 39.34 200
7.6 no 42.74 200
7.8 50 45.17 200
8.0 50 4B.8li 200

Boric Acid. l{CI-N"OH Mixture"

pH
0.2 M HallO" 0.2 M KCI, I 0.2 J1f NaOH. ce. Dil ute to\ ec.
cc.

7.8 50 2.G5 200

8.0 50 4.00 200
8.2 50 5.90 200
8.4 50 8.55 200
8.6 SO 12.00 200
8.8 50 16.40 200
. 9.0 50 21.40 200
9.2 50 26.70 200
9.4 50 32.00 200
9.6 50 30.85 200
9.8 50 40.80 200
10.0 .50 43.90 200

1. Potassium Biphthalate Solution.-Dissolve 40.836 Gm. of

reagent potassium biphthalate in distilled water and dilute to
1,000 cc. to prepare a 0.2 M solution for use in preparing the
buffer solutions.
2. MonopotassiumPhosphate Solution.--Dissolve 27.240 Gm. of
reagent monopotassium phosphate in distilled water and dilute
to 1,000 cc. to make a 0.2 M solutio:t?:: for use in preparing the
buffer solutions.
3. Boric Acid and Potassium Chloride Sollttion.-c-Dissolve
12.404 Gm. of reagent boric acid and 14.912 Gm. of reagent
296 QUANTIT.ATIVE PHARMACEUTICAL CHEMIBTRY

potassium chloride in distilled water and dilute to 1,000 cc.

to prepare a 0.2 M solution of these mixed salts.
4. Potassium Chloride Solution.-Dissolve 14.912 Om. reagent
potassium chloride in distilled water and dilute to 1,000 cc. to
make a 0.2 M solution for use in preparing the buffer solutions.
5. Potassium Biphthalate Solution for Hydrogen Electrode.-
Solution (1) diluted with three volumes of distilled water at
20C. prepares 0.05 M potassium biphthalate solution for check-
ing the hydrogen electrode.
Directions for the pH Measurements.-In the determination
of an unknown pH value, a suitable indicator must first be found.
Only indicators which show an intermediate color between the
extreme acid and alkaline colors can be used.
The first step in the choice of a suitable indicator is the determi-
nation of the approximate pH value of the solution under investi-
gation. A few simple tests will usually supply the necessary
information. Add a drop or two of phenolphthalein T.S. to a
small portion of the solution. If the indicator remains colorless,
the pH of the solution is less than 8.4. A second test is con-
ducted in the same manner, using methyl orange T.S. as the
indicator. If the solution assumes the alkaline color (yellow),
the pH of the solution is greater than 4.4 and lies somewhere
between 4.4 and 8.0. A few more tests with methyl red (pH
interval 4.2 to 6.3), bromthymol blue (6.0 to 7.6), and phenol
red (6.8 to 8.4) will give a close enough approximation of the
pH value to show which indicator may be successfully used in
the determination.
Instead of testing small amounts of the liquid with indicator
solutions, the spot method, using indicator papers, may be
substituted. Also, by using a universal indicator in place of
the several indicator solutions suggested above, some time may
be saved. A so-called unillersal or mixed indicator of this type
can be prepared t;LS follows: Dissolve 0.1 Gm. of phenolphthalein,
0.2 Om. of methyl red, 0.3 Om. of dimethyl-aminoazobenzene,
0.4 Om. of bromthymol blue, and 0.5 Gm. of thymol blue in
500 ee. of absolute alcohol and then add sufficient sodium
hydroxide to produce a yellow color. When 0.1 to 0.5 cc. of this
mixed indicator is added to an unknown solution, the color pro-
duced indicates the approximate pH of the solution as follows:
DETERMINATION OF HYDROGEN ION CONGEN'l'RATION 297
Red color = pH 2 j orange color = pH 4 j yellow color = pH 6;
green color = pH 8; and blue color = pH 10.
When the approximate pH value has been determined and a
suitable indicator agreed upon, a 3, 5, or 10 cc. portion of the
unknown solution (depending upon the amount of the liquid
available), is transferred to a hard, resistant-glass test tube
approximately 15 cm. long and 1.5 cm. 0.5 mm. bore, and a
measured amount of the indicator solution added. As a rule,
0.10 to 0.20 ceo of a 0.05 pel' cent indicator solution, added from
a 1 cc. pipette, graduated to 0.01 ceo per 10 cC. of the solution
being tested, eonstitutes a proper indicator concentration.
Transfer from 4 to 6 portions of the buffer solutions, thepH
values of which overlap that of the unknown solution, to test
tubes and treat in exactly the same way as the solution being
analyzed. The same amounts of indicator must be added to
the unknown and to the buffer solutions. It is also essential
that the test tubes used be of the quality and type already
indicated. The color of the unknown solution is then compared
with the colors of the buffer solutions and the pH value of the
solution thus determined.
In judging the colors, observe them against a white background
with the light transmitted through the whole length of the tube.
A suitable colorimeter may also be used, although it is not neces-
sary in routine work. A sufficient number of reference solutions
must be' taken so that the color of the unknown falls between
two of the series, differing by not more than 0.20 pH. The pH
of the unknown can thus be easily approximated to within 0.1
and with practice to 0.05. With buffer solutions differing by
0.1 pH unit or less, the experimental error can be reduced to
about 0.02 pH.
Other colorimetric methods capable of the same or a greater
degree of accuracy than that officially required may be employed
at the discretion of the operator.
The dissociation constants of indicators hold for aqueous solu-
tions only. Alcohol changes this value and therefore for alcoholic
solutions, a correction factor must be used or the potentiometric
method employed.
Color standards are prepared by placing 10 cc. of a buffer
solution of the proper pH in each of nine Pyrex tubes, adding
298 QUAN1'ITATIVE PHARMA.CEUTICAL CHEMISTRY

0.5 cC. of the indicator solution to each of them, and then sealing
the tubes. Thus in preparing color standards for phenol red
10 cC. of nine buffer mixtures having pH values of 6.8, 7.0, 7.2,
7.4, 7~6, 7.8,8.0,8.2, and 8.4, respectively, are placed in separate
tubes, 0.5 cC. of 0.02 pel' cent phenol reel indicator solution is
added to each tube, and the tnbes are sealed. Each tube is
then labeled with the respective pH value corresponding to
that of the buffer solution which it contains. The tube marked
pH 6.8 will exhibit the yellow color and that marked pH 8.4 will
exhibit the red color of phenol
red, and the tubes with inter-
mediate pH values will show
gradations of color between
'yellow and red.
Reliable color standards
covering the pH range 1.2 to
10.4 for all of the required
indicators, guaranteed for one
year, can be purchased. It is
unnecessary to secure color
standardH for all of the indi-
FIG. 48.-Block comparator.
cators covering the range
pH = 1.2 to. 10.4 unless deter-
minations of pH on a variety of products of widely divergent
hydrogen ion concentration n,re to be made. Thus if a solution
has a pH falling between 6 and 8.4, color standards for brom-
thymol blue and cresol red are all that is necessary.
Color Comparators.-Some type of color comparator must be
used to match the color of the unknown with the color standards.
This is especially true if the sample under examination is at all
turbid or colored. Of the numerOnS devices available, the
LaMotte block comparator (Fig. 48) is best suited for use with
the equipment herein described. This comparator consists of
anyone set of ,color standards such as bromthymol blue; four
test tubes graduated at 10 cc. and of the same bore and wall thick-
ness as the color standard tubes; and a 50 cc. bottle of the corre~
sponcling indicator solution; a tube of distilled water; and a
pipette contained in a wooden case. The top of this case is so
designed that it may be removed and used as a comparator
block.
DETERMINATION OF HYDROGEN ION CONCENTRATION 299

The block comparator is used as follows: Assume that a pre-

liminary test made with various indicators or a mixed incheator
shows that the solution being tested has a pH falling between
pH 6.0 and 7.5. A set of bromthymol blue color standards and
indicator would then serve for the determination. To make the
determination, remove the top of the comparator case (Fig. 48),
and use it as a comparator block. The six holes hack of the slots
in the side of this block are desig-
nated B, A, C and E, D, F, respec- B A c
tively, in Fig. 49.
Fill three of the test tubes to the
10 cc. graduation mark with the
liquid to be tested and place them in
o 0'0
the holes marked B, A, and C. Add
0.5 cc. of bromthymol blue to the
000
tube in A and shake the tube well to E D F
mix the contents. Place the tube con- FIG. 49.-PositioUil of tube~
in the block comparator.
taining distilled 'Ycater in the hole
marked D, and t,vo of the tubes containing color standards
differing only by, 0.2 pH, e.g., 6.8 and 7.0, in the holes E
and F, respectively. Place the comparator block before a
window or other source of light, observe the three pairs
of tubes through the slots, and change the color standards
in E and F, if necessary, until the color of the central pair exactly
matches the color in one of the other pairs or until the color of
the central pair lies between that of the pairs on either side.
If the color of the solution in the central pair matches that of one
of the other pairs, the pH is read off all the color standard with
which the match is obtained. If, however, the color of the solu- ~
tion in the central pair of tubes lies between the colors of the
pairs on either side, the average pH of the latter is taken as the
pH of the sample; e.g., if it lies between 7.0 and 7.2, the value
is taken as 7.1.
The LaMotte roulette comparator (Fig. 50) is designed for
rapid pH determinations with artificial illumination. This com-
parator consists of a stationary base and metal band, and a
wooden drum which revolves inside the band on ball bearings.
Illumination is provided by a 40-watt Mazda lamp attached in
the center of the base. A piece of "Dalite" glass is placed in the
300 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

back of the block between the three test tubes and the color
standards, and a piece of etched glass is placed on the outside
of the block directly over the three slots. This contrivance pro-
vides standard conditions of illumination at all times. Ampoules
of distilled water, bottles of indicator solution, graduated test
tubes, and pipettes are a part of the comparator equipment.
Any three sets of color standards such as chlorphenol red,
pH 5.2 to 6.8; bromthymol blue, pH 6.0 to 7.6; and phenol
red, pH 6.8 to 8.4 are placed in alternate holes in the revolving
drum in the order of their pH. Tubes of the same bore filled
with distilled water are then placed in the vacant holes. There

FrQ. 50.-The rouletto comparator with accessory equipment.

will then be a tube of distilled water beside each tube of color
standard. If the pH of the solution to be tested falls somewhere
within the range of bromthymol blue, pH = 6.0 to 7.6 as deter-
mined by preliminary tests (see page 296), fill three of the test
tubes to the 10 cc. graduation mark with the solution and place
them in the three holes in the block. Add 0.5 cc. of bromthymol
blue to the middle tube and mix it in well by shaking. Turn on
the light and revolve the drum until the bromthymol blue color
standards are directly behind the test sample. While looking
through the slots in the block, gradually shift" the standards by
rotating the drum until the color through the test sample exactly
matches that of one of the tubes on either side of it or lies between
them. The pH value may then be read off directly from the color
'$ta,ndards in the Sfuue manner as with the block comparator.
DETERMINATION Oli' HYDROGEN ION CONCEN'l'RL1TION 301

Exercise 91

Object.-To Determine the pH of Solution of Epinephrine

Hydrochloride, and Syrup of Hydriodic Acid.
Materials Required.-A set of color standards.
Indicator solutions corresponding to t,he color standards.
A color comparator.
40 cc. of solution of epinephrine hyclroehlol'iclc, and syrup of hydriodic
acid.
Procedure.-Determine the pH of each of the respective prepamtions
by the method described for either the block or roulette comparator. Cal-
culate the hydrogen ion concentration of each preparation.

Questiolls and Problems

1. Enumerate several instances in the description of official product8
where such terms as slight[y acid, alkaline to litmus, etc., could be l'eplar.ed
by definite limits of pH to advantage.
2. What advantages does the colorimetric method have that are not
possessed by the hydrogen electrode method of hydrogen ion determination?
3. Name several classes of pharmaceutical preparations of which the pH
cannot be determinc(l colorimetric ally .
4. Ascertain the official pH requirements of water, di8tilled water,
redistilled water, sterilized distilled water, and reagent potassium biphos-
phate. How may the pH limits be determined?
6. What use is made of pH in the determination of the purity of oxygen
and nitrogen monoxide? (See U.S.P.)
6. It is required ill the official standards that fluidextract of aconite be
adjusted to a pH of 2.75 to 3.25 and that tincture of Reonite be adjusted
to a pH of 2.8 to 3.2. If Olle of these preparations had a pH of 4.0, how
would you proceed to adjust it to the official requirements?

Hydrogen Ion Concentration (pH) of Some Official Substances.

The hydrogen ion concentrations, expressed as pH, of the sub-
stances in the following table are not intended to be construed as a
means for the determination of the purity of these substances.
In practice, variations from these figures may frequently be
found, as a slight excess of acid 01' base is, in :ll1any instances,
desirable and even necessary to insure stability and other qualities
in connection with the use of these substances.
When only one figure is given in the table it represents the
theoretical pH, or the one generally agreed upon in the literature.
For the majority of the alkali and alkaline earth salts, an approxi-
mate range is given within which the pH of these substances, as
302 QUANTITATIVE PHARMACEU7'ICAL G'fIEMIS7'RY

TABLE XLII.-pH OF SOlliE OF~'ICIAL SURS'l'ANCES

Subotance ConcentraUon pH

Acid, acetic ......................... . 0.1 M 2.9

Acid, benzoic ....................... . Saturated solution 2.8
Acid, boric ......................... . 0.1 M 5.1
Acid, citric .......................... . 0.1 M 2.1
Acid, hydriodic ...................... . 0.1 M 1.0
Acid, hydrochlmic ................... . 0.1 M 1.0
Acid, hypophosphol'ous ............... . 0.1 M 1.5
Acid, lactic ......................... . 0.1 M 2.4
Acid, nitric ......................... . 0.1 M 1.1
Acid, phosphoric .................... . 0.1 M 1.5
Acid, salicylic ....................... . Sr1tuI'lLted solution 2.4
Acid, sulfuric ....................... . 0.05 M 1.2
Acid, tart!lric ....................... . 0.1.M 1.9
Acid, trichloroacetic .................. . 0.1 M 1.2
Alum (ammonium) ................... . 0.05 M 4.6
Alum (potassium) .................... . 0.1 M 4.2
Ammonium bromicle .................. . 0.1 M 4.6
Ammonium chloride .................. . 0.1 M 4.6
Ammonia water ...................... . 0.1 M 11.3
Apomorphine hydr(.)JJhloridc ........... . 1 in 300 4.8
Arsphenamine ....................... . I in 20 3.0
Atropine ................. ' .......... . Saturated solution 9.5
Atropine sulfate ..................... . 1 in 100 5.4
Caffeine citrated ..................... . 1 in 25 2.3
Caffeine with sodium benzoate ......... . 1 in 25 7.4
Calcium bromide ..................... . 0.2 M 7.0 to 8.0
Calcium chloride ..................... . 0.2.M 6.5 to 7.5
Calcium hydroxide ................... . Sfltuntted solution, 13.5
Calciulll lactate ...................... . 1 in 25 6.0 to 7.0
Cinchonidine sulfate .................. . Sllturated solution 6.4
Cocnine hydrochloride ................ . 0.1 M 4.5
Codeine ............................. . Saturated solution 9.8
Codeine phosphate ........ , .......... . 0.1 M 4.5
Codeine sulfate ...................... . O.lJl.f 5.0
Emetine hydrochloride ................ . 1 in 50 5.6
Ephedrine ........................... . 1 in 200 10.8
Ephedrine hydrochloridr, .............. . 1 in 200 5.1
Homatropine hydrobromide .... " . ; ... . 1 in 100 4.4
Magma magnesia .......... , ......... . ............... . 10.6
Magnesium sulfate ................... . 0.2 M 6.0 to 7.0
Morphine sulfate ..................... . 0.1 M 4.8
Physostigmine salicylate ........... ' ... . 1 in 200 5.8
Piloc!1rpine nitrate ................... . 1 in 100 4.8
DETERMINA.TION OF HYDROGEN ION CONCENTRATION 30J
TABLE XLIL-pH OF SOME OFFICIAL SUBS1'ANCES.-(Continued)

Substance Concentration pH
--------~---I---------
Potassium acetate. . . . . . . . . . . . . . . . . . . .. O. 1 Af 9.7
Potassium bicarbonate....... . . . . . . . . .. 0.1 M 8.2
Potassium bromidc .................... 0.2 M 6.5 to 8.0
Potassium carbonate. . . . . . . . . . . . . . . . .. O. 1 M 11 .6
Potassium hydroxide. . . . . . . . . . . . . . . . .. O. 1 M 13.5
Potassium iodide ...................... 0.2 M 7.0 to 9.0
Potassium nitrate ..................... 0.2 M 6.5 to 7.5
Potassium and sodium tartrate. . . . . . . .. 0.2 M 7.0 to 8.0
Procaine hydrochloride. . . . . . . . . . . . . . .. O. 1 M 6.0
Quinidine sulfatc ........... , . . . . . . . ... 1 in 200 6.4
Quinine... .... .. . . . . . . . . . . . . . . . . . . . .. Saturated solution 8.8.
Quinine bisulfate. . . . . . . . . . . . . . . . . . . . .. 1 in 25 3.5
Quinine dihydrochloride. . . . . . . . . . . . . .. 1 in 25 2.6
Quininehydrobromide..... . . . . . . . . . ... 1 in 25 6.4
Quinine hydrochloride. .. . . . . . . . . . . . . .. 1 in 25 6.4
Quinine sulfate. . . . . . . . . . . . . . . . . . . . . .. Saturated solution 6.2
Quinine and urea hydrochloride. . . . . . . .. 1 in 20 3.1
Sodium acetate... . . . . . . . . . . . . . . . . . . .. 0.1 M 9.7
Sodium benzoate. . . . . . . . . . . . . . . . . . . . .. 0.1 M 8.0
Sodium bicarbonate. . . . . . . . . . . . . . . . . .. O. 1 M 8.2
Sodium biphosphate. . . . . . . . . . . . . . . . . .. O. 1 M 4.5
Sodium bomte ........................ 0.1 M 9.2
Sodium bromidc. . . . . . . . . . . . . . . . . . . . .. O. 2 M 6.5 to 8.0
Sodium cacodylate. . . . . . . . . . . . . . . . . . .. O. 1 M 7.8
Sodium carbonate. . . . . . . . . . . . . . . . . . . .. O. 1 lY[ 11.6
Sodium chloride.. . . . . . . . . . . . . . . . . . . .. 0 . 2 M 6.7to7.3
Sodium hydroxide. . . . . . . . . . . . . . . . . . . .. O. 1 M 13.5
Sodium iodide. . . . . . . . . . . . . . . . . . . . . . .. 0 . 2 M 8.0 to 9.5
Sodium phosphate (dibasic). .. . . . . . . . .. 0.1 M 9.5
Sodium salicylate. . . . . . . . . . . . . . . . . . . .. 0.2 M 5.0 to 6.0
Sodium sulfate ...................... :. 0.2 M 6.0 to 7.5
Sodium thiosulfate. . . . . . . . . . . . . . . . . . .. 0 . 2 M 6.5 to 8.0
Soluble barbital. . . . . . . . . . . . . . . . . . . . . .. O. 1 M 9.4
Strychnine nitrate ...................... 1 in 250 5.7
Strychnine sulfate.. ..... . . . . . . . . . . . . .. 1 in 100 5.5
Theobromine with sodium salicylate..... 1 in 100 10.3

they usually oceur on the market, will fall. Some ueviations from
these values may, however, be expected, as the presence of even
a very slight excess of base or acid in these salts, 01' of carbon
dioxide in their solutions, exercises a pronounced influence upon
the hydrogen ion concentration.
304 QUAN'l'I1'APIVE PHARMACEU'l'ICAL CHEMIS'l'RY

References
1. BRITTON, "Hydrogen Ions," Chapm!ln lind Hall, Ltd., London, 1932.
2. CLARK, "The Determination of Hydrogen Ions," Williams & Wilkins
Press, Baltimore.
8. KaLTHoFF and li'URMAN, "Potentiomctric Titrations," .John Wiley &
Sons, Inc., New York, 1931.
4. Leeds Northrup and Company Bulletins, Leeds Northrup and Company
Philadelphia. .,
 CHAPTER XIX
ELECTROLYTIC METHODS
Quantitative analysis by means of electrolysis, or elcctro-
analysis, as it is more generally called, is usually restricted to the
determination of metals. This method of annJYi:iis is based upon
the fact that an electrical current passed through a solution of
the salt of a metal causes the deposition of the metal, usually
in the elemental state, upon one of the electrodes. The electro-
lytic method may be applied to a number of official substances,
namely, mercury and its salts and the salts of silver, copper, and
zinc.
Electrical Units and Fundamental Laws.-The unit of current
is the ampere, that of resistance is the ohm, and that of difference
of potential or electromotive force is the volt. The ampere is
the strength of current which when passed through a solution of
silver nitrate under certain standard conditions will deposit 0.001118
Gm. of silver per second. The ohm is defined as the 1'esisiance
offered to an unvarying electric current by a column of mercury
106.3 cm. long and 1 sq. mm. in cross-section at OC. The volt
is the electromotive force necessary to fOl'ce a current of one ampel'e
through a resistance of one ohm. The relationship between the
ampere, volt, and ohm is expressed in Ohm's law, viz.: The
strength of an electric current flowing through a cond1tctor is directly
proportional to the difference of potential between the ends of the
conductor and inversely proportional to its resistance. If C repre-
sents the strength of the current in amperes, E the difference of
potential in volts, and R the resistance in ohms, Ohm's law may
be formulated as follows:
C --_
R or, t ranspose d, E1 = CR

The calculation of the amount of metal which will be deposited

du:ring electrolysis in a given period of time is founded on Fara-
day's laws, which may be stated as follows:
305
306 QUAN'l'ITATIVE PHARMACEU'l'ICAL CHEMISTRY

1. The mass of any substance delJOsited at an electrode from a

solution of electrolyte is proportional to the quantity of electricity
which passes through the solution.
2. The amounts of different substances liberated at the electTodes,
when the same quantity of electricity passes through solutions of
different electrolytes, are proportional to the chemically equivalent
weights of the sttbsiances.
From the first law of Faraday it follows that the weight of
.substance deposited from solution during a given period of time
'will be directly proportional to the current strength or nmperage,
Md under a given amperage, will be directly proportionnl to the
time.
The unit quantity of electricity is the coulomb. It may be
defined as the amount of electricity which passes thmugh a conductor
when one ampere flows for one second. It has been found by
experimcnt that 96,500 coulombs of electricity are required to
liberate a gram-equivn,lent weight of any substance at an elec-
trode. The electrochemical equivalent of an element 01' group of
elements is the weight in grams deposited at an electrode by the
passage of one coulomb of electricity. The Blectrochemical
equivalents are proportional to the chemieal equivHlents. Thus
96,500 coulombs of electricity are capable of depositing H/l
= 1.008 Gm. hydrogen: Therefore, 1 coulomb can liberate
1.008 ..;-. 96,500 = 0.0000104463 Gm. hydrogen. This same
quantity of electricity will liberate 35.45 X 0.0000104463 ==
0.00036749 Gm. chlorine, and 107.88 X 0.0000104463 = 0.001118
Gm. silver, and 96,500 coulombs of electricity will deposit
Ag/1 == 107.88 Gm. silver from a solution of a silver salt; Cu/2
== 31.785 Gm. copper from a solution of a cupric salt; and Fe/3
= 18.61 Gm. iron from a solution of a ferric salt. The quantity,
96,500 coulombs, is thus observed to be a unit of quantity in
electrochemical measurements. This unit of quantity is known
as the Faraday, and it is commonly represented by the letter F.
Theory.-Those substances which conduct an electric current
when dissolved in aqueous solution are termed electrolytes.
Electrolytic determinations are based upon the fact that when
a direct current of electricity is passed through a solution of an
electrolyte, some of the negatively charged ions are oxidized to
the elemental state, and some of the positively charged ions are
 ELECTROLYTIC METHODS 307
reduced to the elemental state. This decomposition of an
electrolyte in solution is known as electrolysis, and when the
decomposition of the electrolyte is carried out for purposes of
analysis, the process is called electro-anal:ysis. The conductor
by which the current cnters the solution of electrolyte is called
the anode, and that by which it leaves the solution is known as
the cathode. The anode and cathode are generally spoken of
as positive and negative electrodes, respectively, or simply as
electrodes. The source of current charges the positive elentrodc
(anode) with positive electricity, and this anode attracts the
negatively charged ions (anions), but it repels the positively
charged ions (cations); the latter are attracted by the negatively
charged cathode which repels the negatively charged anion:;
forcing them toward the anode. Since the positively charged
ions gain an electron for each positive charge which they carry
when they come in contact ,vith the cathode, they are reduced;
and since the negatively charged ions lose an electron for each
negative charge which they carry when they come in contact
with the anode, they are oxidized. The respective reductions
and oxidations never take place in the mass of the solution of
electrolyte but only at the electrodes, reduction occurring at the
cathode and oxidation occurring at the anode. Simple metallic
ions (cations) such as copper, mercury, silver, and zinc travel to
the cathode where they are reduced to the metallic state. The
simple anions, such as chlorine, bromine, or iodine travel tr)
the anode and are oxidized to the molecular condition, while
solutions containing hydroxide, sulfate, or nitrate ions liberate
oxygen at the anode. The halogen or oxygen liberated may
escape from the solution or it may act directly or indirectly as an
oxidizing agent.
The various anions are not all oxidized with the same ease,
their relative order being sulfide, iodide, bromide, chloride, and
sulfate, where the first Hamed is the most readily oxidized.
Likewise, the different cations are not all reduced with the same
ease, their relative order being auric, platinic, silver, mercurous,
cupric, arsenous, bismuth, hydrogen, and lead, where the first
named is most readily reduced. The electrolytic method of
analysis is, therefore, a type of oxidation-reduction a.nalysis,
since the oxidation of one species of ion always takes place at the
308 (JUAN'l'ITATIl'E PHARMACEU'l'[CAL CHEMISTRY

anode simultaneously with the reduction of another species of

ion at the cathode.
The minimum voltage necessary to separate ions frorn the charges
which they carry is known as the decomposition voltage. The
decomposition voltages of the ions of qificial substances which
can be assayed by this method are given in the following table:
TABLE XLIII.-DECOMPOSI'lION VOLTAGES

Decomposition
Ion I Metal voltage

Zn++ ............... Zn -0.7581 toO.80

FeH . . . . . . . . . . . . . . Fe -0.441 Lo 0.44
Sn++ ............... Sn -{).lS6 -toCl.14
Pb H .... ; . . . . . . . . Ph -0.122 to 0.12
fI+ ................. H2(Pt) 0.0 to 0.0
Ou H ............. eu +0.3448 to 0.34
Hg+' ............... Hg +0.7986 to 0.80
Ag+ ... : ........... Ag +{) .7995 to 0 . 80
OH-............... 02(Pt) +0.3957 to 0.40
r-.................. 12 (Pt) +0.5357 to O.54
Br-................ Br2(Pt) +1. 0659 to 1. 066
01-................. Cl,(Pt) +1. 3504 to 1. 36
(Pt) = platinum electrode.

Since during electrolysis deposition of the ions occurs at both

electrodes, the voltage required to liberate the ions at both the
anode and cathode must be considQred. For example, when
mercury ions from a solution of mercuric iodide are reduced to
metallic mercury, the decomposition voltage for the mercury ion
is +0.80 while that for the iodide ion is +0.53. Since these
voltages have the same sign, their sum, when added algebraically,
is 0.80 - 0.53 = 0.27. The minimum voltage at which mercury
and iodine can be liberated from mercuric iodide solution is
therefore, 0.27 volt. If zinc chloride is subjected to electrol-
ysis, the respective decomposition voltages for the ions being
-0.76 and +1.36, the decomposition voltage necessary is equal
to their algebraic sum, 2.12 volts.
The difference in the decomposition voltages of mercuric iodide
and zinc chloride solutions make it possible to separat~ mercury
from zinc in a solution containing these salts by maintaining the
 ELECTROLY.TIC ME1'HODS 309

voltage below 2.12 and above 0.27. The greater the difference is
between the decomposition voltages of two metallic ions the easier
it is to separate them by electrolysis. In general, it may be said,
that the decomposition voltages of two substances must differ
by at least 0.2 to 0.3 volt in order to permit their separation by
electrolysis.
Electrolytic methods of assay are not so simple as is implied
in the foregoing examples, since numerous factors complicate
conditions, viz., resistance of the solution, polarization, over-
voltage, reactions at the electrodes, the nature of the solutions
used, the current density, the rate of stirring, the shape and size

FIG. 51.-GaU1I6 anode. FIG. 52.-Spiral anode. FIG. 53.-Spiral anode.

of the electrodes, and the temperature. A discussion of all of

these factors is beyond the scope of the present work; those who
desire a 'more comprehensive knowledge of them should consult
a treatise on physical chemistry.
Apparatus.-The most satisfactory source of current for
electrolytic assays is that furnished by. a lead storage cell or a
number of such cells combined into a battery. Each cell should
supply an electromotive force of about 2 volts so that the con-
nection in series of two or more cells will make it possible to
secure a voltage range of from 2 to 12 volts. The use of the
lighting current is possible provided it is a direct current, but
since the ordinary service voltage is usual1y 110 to 125 volts,
the voltage must be greatly. reduced. An electrical switch is
necessary to make and break the current. The potential differ-
ence between the electrodes is regulated by means of a resistance
310 QUANTITA'l'IVE PHARMACEUTICAL CHEMISTRY

device. A rheostat may be employed, but the ordinary sliding-

contact resistance coil is convenient and satisfactory for most
purposes. The difference of potential between the electrodes
is measured by means of a voltmeter. The amount of current
flowirig . through the electrolytic circuit is measured by an
ammeter: The voltmeter and ammeter employed should be
designed for a limited range of measurement so that they may
be read easily and accurately.
The anode employed in electrolytic determinations usually
consists of a spiral, gauze, or disk of platinum of such construction
that it ean be attached to a. stirrer. Platinum is employed

, ,., ~

. '
:'';:1'. .

;- , ~ '(if' '

f.:)J

FIG. 54.-Pl atin urn dish FIG. 55.-Plutinum FIG. 56.-Platinum

cathode. foil cathode. gauze cathode.

because it, is very resistant to the chemical action of the anions.

Some of the commonly used forms of anodes are illustrated in
Figs. 51; 52, and 53. Although any rotating device may be
employed, a friction drive motor stirrer is very convenient, since
the number of revolutions of the anode per minute can be easily
adjusted.
The material of which the cathode is made depends upon the
metal which'is to be deposited upon it. A platinum dish of
about 200 cc. capacity serves well as cathode, (Fig. 54) in all
of the electrolytic assays of official substances except that of
zinc. In the assay of mercury and its salts and preparations,
however, a mercury cathode cup is recommended. The mercury
cathode cup consists of a glass vessel about 7.5 em. in height
and 4 cm. in diameter with a piece of platinum wire fused into
and through the bottom of the cup and containing about 50 Gm.
 ELECTROLYTIC METHODS 311

_B

FIG. 57.-Diagram of connections fur electrolytic apparatus. a, anode; c.

cathode. A, ammeter; B, battery; V, voltmeter; R, rheostat.

FIG. 58.-Rotating anode and stirring device.

312 QUANTITATIVE PHARMACEU'l'ICAL CHEMISTRY

of mercury. The platinum wire must extend through the glass

wall to make contact with the mercury. In the determination
of zinc electrolytically, either a nickel dish of about 175 cc.
capacity or a platinum dish upon which a thin layer of copper or
silver has been deposited may be employed as the cathode .. The
platinum dish type of cathode can advantageously be replaced
by a hollow cylinder of platinum gauze or foil (Figs. 55 and 56).
These types of cathode are more efficient than the platinum dish
cathode because they present a large surface all parts of which
receive the deposit equally, and because they allow free circula-
tion of the electrolyte. They are also more economical, since
they are light in weight.
The method of connecting up the apparatus is shown in the
diagram (Fig .57).
A convenient rotating anode and stirring device is illustrated
in Fig. 58.
Immediately before starting an assay, both the anode and
cathode should be cleaned by immersing them in a hot solution
consisting of equal parts of concentrated nitric acid and distilled
water. They should then be thoroughly rinsed with distilled
water and ignited to a dull red heat over the blue flame of a gas
burner. This treatment removes any grease or other impurity
present and leaves a clean electrode surface which is essential,
particularly with reference to the cathode, to the production of a
firm, even deposition of metal.

Questions and Problems

1. Define the following terms: ampere, ohm, volt, coulomb, Faraday,

anode, and cathode. .
2. State Ohm's law and Faraday's laws.
3. Show by means bf a diagrnm how the electrolytic apparatus is con-
nected in the electrolytic assay of mercury when a mercury cathode cup
is used.
4. Calculate the decomposition voltage of mercuric chloride.
6. An electric current is passed simultaneously through a series of three
electrolytic cells .containing water acidulated with sulfuric acid, silver
nitrate in sodium cyanide solution, and zinc sulfate in sodium hydroxide
solution, respectively~ How much silver and how much zinc will be depos-
ited if 0.504 Gm. of hydrogen gas measuredl1t OC. and 760 mm. is liberated
from the water?
 ELECTROLY'l'IC .METHODS 313

Exercise 92

Object.-To Assay Copper- Sulfate.

Materials Required.-An electrolytic apparatus.
1 Gm. of copper sulfate.
Hydrogen sulfide T.S.
25 ceo of alcohol.
25 cc. of ether.
15 ee. of concentrated nitric acid.
Procedure.-l, Dissolve from 0.75 to l.0 GIll. of the salt, accurately
weighed, in about 100 co. of distilled water in a weighed platinum dish, add
1 ce. of diluted sulfuric acid, rotate thc anode about 500 revolutions per
minute and pass the current, gradually increasing it to 2.5 amperes and from
5 to 7 volts. Occasionally add a little distilled water to the electrolyte to
keep the deposit of copper covered.

When the cupric sulfate is dissolved in water it ionizes:

CuSOc-+Cu++ + 80 4- -
The copper ions migrate to the cathode where each copper ion
gains two electrons and is reduced to metallic copper. The
sulfate ions migrate to the anode where they react with hydroxyl
ions from the water, forming molecular oxygen and sulfuric
acid:

Each sulfate ion causes two atoms of oxygen combined as

hydroxyl ion to lose an electron and become oxidized to molecular
oxygen. The oxygen is given off at the anode and the hydrogen
ions remaining from the hydroxyl ions unite with the sulfate
ions to form H 2S0 4. If the current employed in the electrolysis
is too strong, however, some sulfate ions may also be discharged:
S04-- + S04-----72803~ + O 2

The 80 a formed then reacts with water to form H 280",:

80 3 + H20~H2804
The diluted sulfuric acid is added because it ionizes strongly
in solution and thereby increases the conductivity of the elec-
trolyte and decreases the resistance of the solution to the passage
of current. It also supplies a high concentration of hydrogen
314 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

ions which tend to prevent the depositiQn of those metals which

are above hydrogen in the electromotive series and to prevent the
oxidation of the deposited copper to the oxide. If the electrolysis
of copper suJfate solution is carried out without the addition of
sulfuric acid, the CQPper deposit is spongy and non-adherent,
but when sulfuric acid is added, the deposit is compact and
adherent.
It has been found by experiment that vigorous stirring of the
electrolyte greatly reduces the time required to carry out an
electrolytic determination, since it brings the ions in contact with
the electrodes. The rotating anode serves as a stirrer.
Small amounts of distilled water are added from time to time to
cover and thereby prevent the oxidation of the freshly deposited
copper by the atmospheric oxygen.

2. When the copper is all depositcd. (usually in about 20 min., to be

ascertained by wHhdrawing a little of the electrolyte [tnd testing it with
hydrogen sulfide T.S.), stop the rotation, and wash the deposit with dis-
tilled water by llleans of a siphon without interrupting the current until the
current drops to nearly zero.

The complete deposition of the copper is indicated by the

disappearance of the characteristic color from the solution.
When the solution is colorless, transfer a drop of it to a depression
in a porcelain spot plate by means of .a glass stirring rod and
add a drop of hydrogen sulfide T .S. If the copper has not all
been deposited, a brownish-black precipitate of cupric sulfide
will form:

If the electrolysis is found to be complete, stop the rotation of

the anode but do not stop the flow of current. Siphon the acid
electrolyte liquor from the cathode cup, but keep the dish filled
by the simultaneous addition of distilled water from a wash
bottle. About 500 cc. of water must usually be employed before
the ammeter will indica;te that practically nO current is "flowing.
The current should not be stopped while washing the acid elec-
trolyte liquor from the cathode, since the deposited copper is
slightly solu.ble in i;)tl'ong acids.
 ELECTROLYTIC ME'l'HODS 31,15

3. Remove the dish, wash again with distilled water hy decantation,

followed by :t little alcohol and ether, successively, dry. for a few minutes
only, at from 80 to 100C., and weigh.

After stopping the current, detach the cathode cup and wash it
thoroughly with water, alcohol, and ether successively, dry it in
an electric oven or air bath for 2 or 3 min. at 80 to 100C.; allow
the cathode to cool in a desiccator, and weigh it. The operations
of drying and weighing should be performed as quiekly as possi-
ble, since the copper is subject to oxidation upon exposure to air.
After weighing the cathode with the copper deposit, dissolve the
deposit completely by treating it with two successive portions
of about 10 cc. of concentrated nitric acid mixed with about
10 cc. of water using gentle heat to effect solution. Wash the
electrode successively with distilled water, alcohol, and ether
and dry again at 80 to 100C., allow it to cool in a desiccator,
and weigh it again. The difference between the weight of the
cathode and the weight of the cathode with the copper. deposit
is equal to the weight of copper deposited. The cathode may
be weighed before the electrolysis is started and again after the
deposition of the copper is complete, but this procedure leads to
a slight inaccuraey since the anode dissolves slightly during the
electrolysis and the dissolved platinum is plated out on the
cathode.
Calculate the percentage Cu, CUS04, and CuS04.5H~O to
which the sample assayed corresponds.
Questions and Problems
1. Explain how oxidat.ion and reduction are involved in the assay of copper
sulfate.
2. Why is the electrolyte acidulated with sulfuric acid?
3. Why is the solution of electrolyte stirred?
4. Why should the current not be stopped until the acid electrolyte has
been washed out of the eathoele dish?
5. How many coulombs of electricity would bc required to completely
deposit the copper from 0.6 Gm. of pure CuS01.5H 20?

Exercise 93
Object.-Assay of Mercuric Chloride.
Materials Required.-An electrolytic apparatus with a mercury cathode
cup.
316 QUAN1'1'1'ATIVE PHARMACEUTICAL CHEMISTRY

0.4 Gm. of mercuric chloride.

1 cc. of concentrated nitric acid.
1 ce. of hydrogen sulfide T.S.
10 ce. of toluene.
25 ce. of alcohol.
25 ce. of ether.
Procedure.-l. Dissolve from 0.3 to 0.4 Gm. of the salt, accurately
weighed, in 10 ce. of distilled water and transfer the solution to the cathode
cup, which has been previously weighed with its metallic mercury.

It is essential that the mercury cathode cup be thoroughly

cleaned. Before using, the cup, emptied of its mercury, should
be washed with concentrated nitric acid to dissolve the film of
amalgam which forms upon the platinum wire. The nitric acid
is removed by rinsing the cup successively with water, alcohol,
and ether and the cup is dried. The mel'Cluy cathode cup should
contain about 50 to 60 cc, of pure mercury. If the mercury
cathode is properly prepared, it may be used for a number of
determinations.
2. Dilute the liquid to about 20 ce. with distilled water, add 1 cc. of
nitric add, which has been diluted with an equal volume of distilled water,
and 10 cc. of toluene, and pass through the solution a current of from 1 to
3 amperes and from 8 to 12 volts, stirring the solution by rotating the !tnode
from 500 to 600 revolutions pel' minute.
After weighing the mercury cathode cup accurately, attach it to
the electrolytic apparatus, introduce the solution of mercuric
chloride into it, and wash the vessel in which t,he solution was
prepared with two 5 cc. portions of distilled water, adding each
of the wash solutions to that contained in the cathode cup. One
cubic centimeter of nitric acid is added to increase the conduc-
tivity of the electrolyte. The toluene is added to absorb the
liberated chlorine gas and prevent the latter from exerting a
solvent action on the platinum anode. After starting the rota-
tion of the anode, turn on the current.
3. After 20 min" when the mercury is all removed from solution (to be
ascertained by removing a few drops of the solution and testing with hydro-
gen sulfide T.S.), wash with distilled water, with the aid of a siphon and
without. interrupting the current, until the current drops to' nearly zero.
Remove the cathode cup, wash the mercury with alcohol, then with ether,
remove most of the remaining ether with filter paper, dry over sulfulic acid,
and' weigh. The inorease in weight of the cathode cup represents the
amount of mercury present in the quantity of the salt taken.
 ELECTROLYTIC METHODS 317

The method of testing the electrolyte for completion of the

electrolysis and the procedure employed in washing out. the
cathode cup have been deseribed under the assay of copper
sulfate (page 313). The acid electrolyte liquor must be removed
before the current is stopped, sinee otherwise the acid will act
upon the mercury causing re-solution of some of the metal.
Calculate the per cent of mercury in and the purity of the
mercuric chloride assayed.

Questions and Problems

1. Explain the oxidation-reduction reactions which oeellr during the
electrolytic assay of mr;reuric chloride in terms of the i~nic theory.
2. What substances other than toluene might be used to absorb the
chlorine liberated at the anode?
3. What assays of other official mereury salts may be performed electro-
lytically?
4. An unknown preparation of mercur~r weighing 0.75 Gm. when assayed
electrolytically yielded 0.2640 Om. of mercury. To what per (:ent of each
of the following in the sample would the amount of rnercury found curre-
spond: HgCl, HgCb, and HgI 2?

Other Electrolytic Assays.-The general prineiples involved in

all assays by electrolytie methods are similar to those applied in
the'two preceding exercises. Some of the following procedures
require explanation, however.
The electrolytic assay of the silver compounds offers 110 special
advantage over the gravimetric chloride precipitation method.
The silver is deposited from an electrolyte containing sodium
cyanide. In the case of silver nitrate, for example, the alkali
cyanide reacts 'with the silver salt to form a double cyanide:
AgNO a + NaCN--..?AgCN + NaNOs
AgCN + NaCN--..?AgNa(CNh
The double cyanide may be represented to ionize as follows:
AgNa(CN)2--..?Ag+ + Na+ + 2CN-
The presence of the alkali cyanide increases the conductivity of
the electrolyte and, as has been found by experiment, causes the
formation of a firlll adherent deposit of silver at the cathode
whereas in the absence of the cyanide a spongy, non-adherent
deposit is frequently obtained.
318 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The mercury compounds insoluble in water for whieh elec-

trolytic assays may be used, such as mercurous chloride, mercu-
rous iodide, mercuric salicylate, and ammoniated mercury, are
electrolyzed from a solution of sodium sulfide, because mercury
sulfide is soluble in the presence of an excess of strong alkali
Rulfide:

The complete deposition of mercury is determined by adding a

few drops of ammonium chloride to a' drop of the electrolyte
contained in a depression of a spot plate. The ammonium
chloride reacts with the sodium sulfide forming ammonium
sulfide in which mercury sulfide is insoluble:

Mercury, if present in the electrolyte, is deposited as a black

precipitate. When the deposition of mercury from the cyanide
solution is complete, the mercury in the cathode cup is washed
with 2 to 3 per cent acetic acid solution to remove the sodium
sulfide which is absorbed or dissolved in small amounts at the
contact surface between the mercury and eleetrolyte:

The deposition of zinc from an electrolyte has been found to

take place best when a nickel cathode is employed and when the
deposition is effected from an electrolyte made alkaline with
sodium hydroxide. The reactions involved in the assay of zinc
chloride are typical of those which occur in all of the electrolytic
determinations of zinc. These reactions may be represented as
follows:

ZnCb +2NaOH~Zn(OHh + 2NaCI

Zn(OHh + NaOH~Zn(OH)ONa + H 2 0
Zn(OH)2~Zn++ + 20H-

The rate at which the zinc is deposited is accelerated by heating

the solution of electrolyte, since zinc deposits rather slowly and
heat increases the speed of the ions.
 ELEC'1'IWLYTIC METHODS 319
TABLE XLIV.-SOME OFFICIAL SUBSTANCES WHICH MAY BE ASSAYED BY
ELlllCTllOLYTW METHODS

({nUllS
equivalent
Amount AIlIpcl'-
Substance Voltage to 10m.
used, age
used of
grams used
Dl'PDBj t l'l)
metal
-~

U.S.P.
Copper sulfate ............ 0.75 to 1.0 2.5 5 to 7 3.9281
Copper sulfate, anhydroUl:; 0.50 to 0.75 2.5 5 to 7 2.5112
Mercury ................. 0.2 to 0.3 1. 5 to 2 7 to 10 l.0000
Mercury, ammoniated ..... 0.5 to 0.6 2 to 3 7lo 10 1.2566
Mercllry bichloride ....... 0.3 to 0.4 1 to 3 8 to 12 1.3535
Mercuric iodide, yellow .... 1.0 to 1.2 2 to 3 7 to 10 2.2654
Mercuric oxide, yellow .... 0.2 to 0.3 1.5 to 2 7 to 10 1.0798
Mercuric salicylate ........ 0.7 to 0.8 2 to 3 7 to 10 l.1798
Mercurous chloride, mild. . 0.5 to 0.6 2 to 3 7 to 10 1.1768
Mercurous iodide, yellow .. 0.7 to 0.8 2 to 3 7 to 10 1.6327
Mercury with ohalk ... " .. 0.6 to 0.8 1. 5 to 2 7 to 10 1.0000
Silver nitrate ............. 0.3 to 0.5 2.5 5 to 7 1.5748
Silver nitrate, toughened... 0.3 to 0.5 2.5 5 t9 7 1.5748
Zinc acetate .............. 0.5 4 to 5 5 to 6 3.3570
Zinc chloride ............. 0.3 4 to 5 5 to 6 2.0849
Zinc oxide ................ 0.2 4 to 5 5 to 6 1.2448
Zinc sulfate .............. 0.6 4 to 5 5 to 6 4.3988
Zinc slllfate, anhydrollS .... 0.4 4 to 5 5 to 6 2.4695
N.F.
Mercuric iodide, red ....... 1.0 to 1.2 2 to 3 7 to 10 2.2654
J'.r.d ____-... 0.2 tDO.3
M'J'!wyjg !w.WI',- l.OtD2 7 tD 10 l.0798
Zinc phenolsulfOllate ...... 1.2 4 to 5 5 to 6 8.5011
 PART III
SPECIAL METHODS USED IN OFFICIAL
PHARMACEUTICAL ANALYSES
Quantitative analyses of crude drugs and of the products
derived from them are made to establish purity or to determine
the amount of therapeutically active constituents prcsent for
the purpose of standardization. The special methods employed
in the analyses of this type may be classified as follows:
Chemical methods such as those employed in the determination
of ash, moisture, crude fiber, extractive obtained with different
solvents, the estimation of alkaloidal content, etc.
Physiological methods or those in which the effects upon
animals or animal tissues are measured and which are employed
in the absence of satisfactory chemical methods for standllrdiza-
tion. The official drugs assayed by physiological methods are:
U.S.P. aconite, tincture of aconite, cod liver oil, 110n-destearillated
cod liver oil, digitalis, digitalis powder, tincture of digitalis,
solution of epinephrine, solution of irradiated ergosterol, ergot,
fluidextract of ergot, extract of liver, solution of liver, purified
solution of liver, solution of parathyroid, solution of posterior
pituitary, stomach, and strophanthin and tests for the toxicity
and purity of arsphenamine, neoarsphenamine, and tryparsamide;
N.F. fluidextract of aconite, convallaria root, fluidextract of
convallaria root, ampuls of epinephrine hydrochloride, extract of
ergot, ampuls of posterior pituitary, fluidextract of squill, stro-
phanthus, tincture of strophanthus, and suprarenal. Physio-
logical methods require special apparatus for their performance
and a technique which is not chemical in nature; hence, a dis-
cussion of them does not fall within the scope of this text.
 CHAPTER XX
ASH AND MOISTURE DETERMINATIONS
Ash Content.-The ash content of a crude drug is generally
taken to be the residue remaining after incineration. It usually
represents the inorganic salts naturally occurring in the drug and
adhering to it, but it may also include inorganic matter added
for the purposes of adulteration. There is a considerable differ-
ence in the ash content of different drugs, but the difference varies
within narrow limits in the case of the same individual, hence an
ash determination furnishes a basis of judging the identity and
cleanliness of a drug and gives information relative to its adultera-
tion with inorganic matter. Ash standards have been established
for a number of the official drugs; usually these standards set a
maximum limit on the total ash or on the acid-insoluble ash
permitted. The total ash is the residue remaining after incinera-
tion, while the acid-insoluble ash is that part of the total ash
which is insoluble in diluted hydrochloric acid. The Pharma-
copoeia specifies that all vegetable drugs shall be separated by
mechanical means in so far as possible from lumps of dirt or other
foreign inorganic matter before they are ground or powdered.
When no ash standard is fixed for a given drug by the Pharma-
copoeia or N !1tional Formulary, the amount of foreign inorganic
matter estimated as acid-insoluble ash must not exceed 2 per
cent of the weight of the drug.
The ash 01' residue yielded by an organic chemical compound
is as a rule a measure 6f the amount of inorganic matter present as
impurity. In most cases, the inorganic matter is present in
small amounts which are difficult to remove in the purification
process and which are not objectionable if only traces are present.
The careful control of temperature is the most important
analytical factor to regulate in making ash determinations. All
determinations should be made in such a manner as to duplicate
in so far as possible the conditions under which standards are
established. When an electric furnace is used for ignitions, the
323
324 QUAN'l'ITATIVE PHA.RMAC'EU'l'ICAL ClJEMIS'l'RY

following temperature equivalents which are only approximate

will be found of value: Very dull red heat = .500 to 5500.;
dull red heat = 550 to 7000. ; bright red heat = 800 to 10000. j
yellow red heat = 1000 to 12000.; and white heat = 1200 to
16000. Practically all of these temperatures are within the
limits of the ordinary BUllsen burner depending on the construc-
tion of the-burner and the nature of the gas burned. When it
is considered that calcium carbonate is converted to calcium
oxide at about 8250., that calcium phosphate is converted to
calcium pyrophosphate at about 15500., that lithium carbonate
forms lithium oxide at about 6000., and that potassium chloride
sublimes at about 1,5000., it becomes apparent that ignitions
should be carried out at low temperatures if possible.
Exercise 94
. Object.-To Determine the Total and Acid-insoluble Ash
Content of Digitalis Leaf.
Materials Required.-2 to 4 Gm. of the ground drug.
2 ashless filter papers.
Procedure.-l. "Accurately weigh a quantity of the ground drug, repre-
senting from 2 to 4 Gm. of the air-dried material, in a tl1l'ed crucible and
ineinemtc at a low temperature, not to exceed very dull redness, until free
from carbon, and determine the weight of the ash."

The crucible in which the drug is to be ignited should be heated

to dull redness until of constant weight, cooled in a desiccator,
and weighed. This procedure drives off the moisture and the
absorbed gases which are present on the surface of the crucible.
The drug is incinerated at a temperature not to exceed a dull
redness. Very often, the ash consists largely of ca-lcium carbon-
ate. formed from the calcium oxalate contained in the drug.
When the incinemtion is carried out at a dull red heat, the calcium
oxalate is converted into carbonate, but when the incineration
is carried out at a bright red hea-t, varying amounts of the
carbonate are converted into the oxide with consequent variable
results. If the ash contains alkali chlorides, which it frequently
does, there may be a- loss of some chloride by volatilization when
a high ignition temperature is used.
t .2. "If a carbon-free ash cannot be obtained ill this way, exhaust the
charred mass with hot distilled wat.er, collect .t.he inlloluble residue on an
 ASH AND MOISTURE DEl'BRMINA.'l'IONS 825

ashless filter paper, incinerate the residue and filter paper until the ll~h is
white or nearly so, then add the filtrate, evaporate it to dryness and heat the
whole to a low redness. If a ca.rbon-frec aHh cannot be obtained in this way
cool the crucible, add 15 ee. of alcohol, break up the a~h with fI glasi'; ro~l:
burn off the alcohol, and again heat the whole to a low redncHs. Finally
determine the weight of the ash. Calculate the percentage of total ash from
the weight of the drug taken."

When the ignition residue is not heated above dull redness, it

is frequently very difficult to burn off all of the carbon, some of
which may become surrounded by fused inorganic matter thereby
keeping it from coming in contact with the oxygen of the air.
In such cases, the residue is treated with hot distilled water to
dissolve most of the inorganic matter, and the insoluble portion
is collected on an ashless filter and incinerated. The filtrate is
then added to this residue, evaporated, and the whole is ignited
to burn off any co1l9idal carbon which may have passed into the
filtrate. Occasionally, it may be necessary to treat the residual
mas's with alcohol to disintegrate the fused lump, especially in the
ignition of resinous drugs.
The weight of the crucible and ash minus the weight of the
crucible gives the weight of total ash obtained from the sample
of drug taken. This total ash, which usually contains carbonates,
phosphates, sulfates, chlorides, oxides, etc., of calcium, magne-
shun, potassium, sodium, aluminum, iron, and other metallic
elements, does not necessarily represent all of the inorganic con-
stituents of the drug, since ammonium salts, some alkali iodides
and nitrates, etc., are volatilized or converted into carbonates, etc.
3. "Boil the ash obtained with 25 cc. of diluted hydrochlorie acid for
five minutes, collect the insoluble matter in a Gooch erucible or on an ashless
filter, wash with hot distilled water, ignite and weigh. Determine the per
cent of ucid-insoluble ash calculated from the weight of drug taken."

The diluted hydrochloric acid dissolves the calcium carbonate,

alkali chlorides, etc., leaving an acid-insoluble residue which
consists almost entirely of silica derived from the soil adhering
to the drug.
Questions and Problems
1. From what different sources may the ash of a drug be derived 'I
21 When no specific ash limit is set by the official standards, what per
cent of acid-insoluble ash is the drug permitted to contain?
326 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

3. Why should the incineration residue not be heated above a dull redness?
4. Name the important inorganic phnt constituents which will not be
found in the ash.
5. What does the quantity of acid-insoluble ash in a drug indicate?

The ash content of chemicals is determined by ignition to

dull redness in the same manner as in the determination of the
ash content of a crude drug.
It is economically impracticable to make ash determinations
on largc samples of expensive chemicals. However, it is neces-
sary to control the amount of inorganic matter which may be
contained in such substances as alkaloidal salts, especially when
the substance is intended for hypodermic administration. For
this reason the official standards usually require that small
amounts of expensive substances be used and that the ash yielded
be negligible. The term negligible is defined as "a quantity
not exceeding 0.0005 Gm." A number of official substances,
with the quantity in Grams in parcntheses, required to yield a
negligible amount of residue are: U.S.P. (0.1) atropine, (0.1)
atropine sulfate, . (0.5) barbital, (0.5) carbromal, (0.5) cocaine,
(0.5) cocaine hydrochloride, (0.5) codeine, (0.5) codeine sul-
fate, (0.1) colchicine, (0.2) emetine hydrochloride, (0.1)
epinephrine, (0.2) ethylmorphine hydrochloride, (0.5) eucaine
hydrochloride, (0.1) homatropine hydrobromide, (0.5) morphine
sulfate, (0.2) pelleticrine tannate, (0.5) phenobarbital, (0.1)
physostigmine salicylate, (0.1) pilocarpine nitrate, (0.5) procaine
hydrochloride, (0.1) scopolamine hydrobromide, (0.1) strophan-
thin, and (0.1) theophylline; N.F. (0.1) cotarnine chloride, (0.1)
hydrastine hydrochloride, (0.5) morphine hydrochloride, and
(0.1) pilocarpine hydrochloride.
The ash limits of most of the official vegetable drugs and
chemicals are indicated in Table XLV. The per cent of
ash given is the maximum permitted for each substance. Those
vegetable drugs for which a maximum limit of 2 per cent of acid-
insoluble ash is permitted are not included.
Residue limits are given in the official standards for a number
of solvents and preparations. The residue is determined by
evaporation and drying to constant weight at a specified tempera-
ture or by evaporation and ignition of the residue as indicated
in Table XLVI, page 329.
 ASH AND MOISTURE DETERMINATIONS 327

TAllLE XLV.-OFFICIAL SUBSTANCES WI'l''H ASH LIMITS

Acid- Acid-
Total inslJlu~ Total insolu-
ll8h, ble ash, ble
Substance Substance
pcr ash, per ash.
cent per cent per
cent cent

--- ---
U.S.P. U.S.P.
Acacia ........ " ....... 4.0 0.5 Digitalis ...... " ....... 5.0
Acetanilid .............. 0.05 Elaterin ....... " ....... 0.1
Acetophenetidin ......... 0.05 Ephedrine .............. 0.1
Acid, acetylsalicylic ..... 0.05 Ephedrine hydrochloride. 0.1
Acid, Ilcetyltannic ....... 0.3 Ephedline sulfate ....... 0.1
Acid, benzoic ........... 0.0.5 Ethyillminobcnzoate ... 0.1
Acid, citric ...... " ..... 0.05 Glucose ....... " ....... 0.5
Acid Illctic .............. 0.12 Glycerin ............... 0.007
Acid, oleic .............. 0.10 Glycyrrhiza ....... " .... 2.5
Acid, salicylic .... '" .... 0.0.5 Glycyrrhiza, extract of ... 8.0
Acid tannic ............
t 0.5 Guaiacol. .............. 0.1
Acid. tartaric ........... 0.05 Hyoscyamus ............ 12.0
Acid. trichloroacetic ..... 0.05 Iodoform ............. 0.2
Agar .... "." .......... 1.0 Iodine ................ 0.05
Albumin tannate ........ 0.3 Lactose ................ 0.1
Aloe ................... 4.0 Menthol. .............. 0.05
Aloin .................. 0.5 Mercuric chloride 300(: . 0.1
Aminopyrine ............ 0.1 Mercuric oxid~} yellow ... 0.2
Ammonium benzonte .... 0.05 Mercuric BtlUcyllite .... , . 0.2
Ammonium bromide ..... 0.05 MercurOILq chloride ...... 0.1
Ammonium carbonate ... 0.05 Mercurous iodide, yellow 0.2
Amlnonium ch1oriue ..... 0.1 Mercury ............... 0.02
Ammonium salicylate .... 0.05 Mercury, n.mmonintcd ... 0.2
Antipyrine ............. 0.1 Mercury succinimide .... 0.1
Asafoetida .............. 15.0 Methenamine ........ , .. 0.05
Aspidium ............... 3.0 Methylthionine chloride. 1.0
Belladonna leaf ......... 3.0 Myristica .............. 0.5
Belladonna root ......... 4.0 Myrrh ................. 4.0
Benzoin, Sumatra, ... , .. 1.0 Paraffin, chloriu!Lted ..... 0.1
Benzoin, Siam .......... 0.5 Petrolatum ............. 0.05
Betanaphthol. .......... 0.05 Petrolatum, white ....... 0.05
Calieine ................ 0.05 Phenacaine hydroohloride 0.1
Calieine, oitr!Lted ........ 0.1 Pheno]. ................ 0.05
Campbor .............. 0.05 Phenol. liquefied ........ 0.05
CannllbiB ............... 5.0 Phenolphthalein ........ 0.013
Capsicum .............. 1.25 Phenolsulfonphthalein ... 0.2
Cardamom seed ....... ' . 5.0 Phenyl salicylate ........ 0.05
Caraway .............. 1.5 Pine tar ................ 0.25
Charcoal, aotivated ..... 4.0 Prepared chalk ......... 2.D
Chloral hydrate ......... 0.05 Pyrogallol. ............. 0.1
Chlorobutanol. ......... 0.1 Quinidine sulfate ..... 0.1
Chrysarobin ............ 0.25 Quinine ............... 0.1
Clove ............ , ..... 0.75 Quinine ethyl carbonate. 0.2
Cotton, purified ......... 0.2 Quinine and urea hydro-
Creosote carbonate ...... 0.1 chloride .............. 0.05
Dextrose ............. ,. 0.1 Quinine bisl,lliate ...... : . 0.05
328 QUANTITATIVE PHARMACEUTICAL CHEMIHTRY

TABLE XLV.-OFFICIAL SUBSTANCES Wl'I'H ASH LIM1Ts.-(Contin1ted)

Acid- Acid-
Total insolu- Total inSDlu-
aRh, ble nsh, ble
Substance Substance
per ash, per ash,
cent per cent per
""'It cent
- - 1 1 - - - - - - - - ---- - - -
U.S.P. N.F.
Quinine dihydrochloridc. 0.05 Colocynth ............. . 6.0
Quinine sulfate ......... . 0.05 Convallaria root. , ..... . 6.0
Resin of pOllophyllum .. . 1.5 Coriander ............. . 1.5
Resorcinol. ............ . 0.05 Corpus luteum. . . . . . . . . . 6. a
Resin ... ' ............. . 0.05 Crocus................. 7.5 1.0
Saccharin, 80luble ...... . 0.5 Cudbear .... '. . . . .. .... 12. a
SantolliJl ............. " 0.1 Dnlniana . .... , ... , .... . 4.0
SllrBapal'illl,., l\1:exicull .. . . 4.0 Euonymus. , .......... . 4.0
Senna ................. . 3.0 Euphorbia ............. . 3.0
Serpentaria .... " ...... 10.0 Fennel ....... , ...... .. . 1.5
Stllrch ................ . 0.5 Gambir ............... . 0:5
Stramonium ........... . 4.0 Galllboge .............. . 1.0
Strychnine sulfate ...... . 0.1 Guaiacol carbonate. . . . . . 0.1
Sucrose .. ...... , ...... . 0.05 Guurnna ...... ........ . 0.5
Sulfonethylmethane .... . 0.05 HuIIlulus .............. . 5.0
SulluI', precipibderl ..... . 0.3 Hydrastis ........... " . 3.0
Sulfur, sublimed ........ . 0.5 Ichthl\mmol. ........ , . . 0 . 5
Sulfur, washed ......... . 0.3 Ipomoea ....... '" . " ., 3.0
Terpin hydrate ......... . 0.05 Iris ................... . 1.0
Theophylline with eth- Jal<Lp ................. . 0.5
ylene diamine ........ . 0.1 Kalll!lla ............... . 0.0
Thymol. .............. . 0.05 Kola .................. . 0.5
Thymol iodide ......... . 1.5 Leptandm. , ...... , .... . fl. 0
Val.rian .............. 10.0 Lobelia ............... . 5.0
Vanillin .............. .. 0.05 Lupulin ............... . 10.0
Veratrum viride ........ 4.0 Mastic .. .... , .. , ...... . 0.25
Wool lnt .............. . 0.1 Mo.tricaria , ........... . 4.0
N.F. Mullein leaves ......... . 4.0
Acid, gallic ............ . 0.1 Ovary and Qva.rian resi-
Aletris ................ . 10.0 due ................. . 7.0
Animal charcoal, purified 4.0 Pimento. .............. . 0.4
Anise ................. . 1.5 Pituitn.ry, ante rio)' ..... . 7.0
Areca ................ 2.5 Pituitary, whole ....... . 7.0
Brucine sulfate ......... . 0.1 Plantago Beed .......... . 4.0 1.0
Buchu ................ . 1.0 Poplar bud ............ . 1.0
C"'lamus .............. . G.O 0.5 Qul1l!Sil1 ............... . 0.5
Calumba .............. . 2.5 Resin, iponlOClt ... , .... . 0.5
Camphor ,monobromated 0.05 ROBe ................ 1.0
C..ramel. ............. .. 8.0 Salvia ................ 10.0 1.0
Carmine .............. . 12.0 Sassafras .............. . 5.0
Cauolphyllum ......... 4.0 Sassafras pi th. . . . . . . . . . 0.5
Celery fruit ............ . 3.0 Strychnine ............ . 0.1
Chlorthymol .... , ...... 0.05 SulfoliemetbaJ1e ........ . 0.05
Cimicifuga. ........... .. 4.0 Suprarenaf............ 7.0
Cinohonine sulfate ..... . 0.1 Taraxacum .......... '" 4.0
Cinchonidine Bullate .... . 0.1 Thyme ................ . 4.0
Cinchophen ........... . 0.25 Triticum .............. . 3.0
Coal tar ............. .. 2.0 UlmuB ................ . 1.0
Colchicum corm ....... . 0.5 Viburnum prunif.olium .. . a.o
 ASH AND MOISTURE DETERMINATIONS 320
TABLE XLVI.-'-SOME OFFICIAL SUBSTANCES WITH RESIDUE REQUIREMENTS

Amount ReHiduc Oftlcial

Substance used, Gm. dried tt1; l'equirc-
or cc. c. mont, Gill.
----
U.S.P.
Acetone ................ " " ...... 50 100 0.002
Acid, acetic ....................... 20 100 0.002
Acid, diluted hydriodic ............. 5 Ignite 0.100
Acid, hydrochloric ................. 10 Ignit.e 0.002
Acid, nitric ....................... 30 Ignite 0.002
Acid, sulfuric ..................... 2 Ignite 0.001
Alcohol .................... , ...... 40 100 0.001
Benzin, petroleum ................. 50 40 0.001
Ether .................. " ........ 50 100 0.001
Ethyl oxide ....................... 50 100 0.001
Water,
ammonitl ....................... 10 100 0.002
distilled ...................... , , 100 100 0,001
distilled, lltel'ilizcd ... , .. , . , .... , . 100 100 0.001
orange flo\', er .... , ..... , ........ 100 100 0.001
rose, stronger ... , .... , , ......... 100 100 0.001
N.F.
Antis!lptic solution ........... ' ..... ;to 100 0.184
Water ............................ 100 100 0.030
Water, hamamelis .... , .. , ..... ,. ,. 100 100 0.025
Water, redistilled ..... , ......... , , . 100 100 0.0005

Moisture Content.-The moisture content of crude drugs is

somewhat variable, since most vegetable drugs are more or less
hygroscopic. Some drugs take up as much as 5 per cent of their
air-dried weight when stOl'ed in a humid atmosphere. Generally,
assays are made to determine the proportion o.f active constitu-
ents contained in a drug in the condition in which it is purchased
or used,. In ,accurate scientific work, however, and in those
cases where the drug is to be sold with a guaranteed assay, the
per cent ,of active constituent must be calculated on the basis
of the moisture-free drug.
The method most, commonly followed in determining the mois-
ture 'content of a drug is to heat it at 100"'0. in an oven until
the weight becomes constant. Since, however, most vegetable
drugs contain variable amounts of volatile substances other than
330 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

water, such as essential oils, ethers, and esters, amines and

alkaloids in some instances, this method is not generally appli-
cable except for rough estimations. When the drug contains
volatile matter other than water, the U.S.P. and N.F. directions
require that the volatile ether-soluble extractive shall be deter-
lllined and the weight of the latter subtracted from the weight
lost by the drug upon drying, the difference to be taken as the
moisture content of the drug. Even this procedure is not free
from error. Since water is slightly miscible with ether, some of
the moisture contained in thc drug is extracted by the anhydrous
ether and determined as volatile ether-soluble extractive. A
better method, which was developed by the U. S. Forestry
Service, is commonly known as the xylene method, although
kerosene, toluene, and other organic solvents have been employed
in the place of xylene. A modification of this method is official
as the moisture method by toluene distillation. It is the method
specified for the determination of the moisture content of thyroid
and is generally applicable to moisture determinations in glandu-
lar products and vegetable drugs containing 2 or more per cent
of moisture. This method has the disadvantage that a com-
paratively large amount of drug, in some eases 50 to. 100 Gm.,
must be used for the determination in order to secure a volume
of water that can be measured conveniently without considerable
error.
Most of the official chemical compounds are relatively stable
in respect to their moisture content at room temperatures, i.e.,
they neither gain nor lose more than slight traces of water. Some
of them are hygroscopic and absorb water readily, e.g., aluminum
sulfate. Others lose a part or all of their water of crystallization
unless properly preserved, e.g., copper sulfate. Control of the
amount of water contained in chemicals is important in making
solutions of definite concentration and in the determination of
the dosage of those substances used as medicaments.
The moisture content of chemicals is determined by drying
them to constant weight, at a specified temperature. In some
cases, the U.S.P. and N.F. specify the allowable pel' cent of
moisture in a chemical; when no limit is specified, 5 per cent of
moisture is permitted. For certain chemicals, a limit of toleranoe
for loss of water of crystallization has been adopted,
 A8H AND MOISTURE DETERMINATIONS 331

Exercise 95

Object.-To Determine the Moisture Content of Acacia.

Materials Required.-l0 Gm. of Acacia.
Procedure.-l. Powder thc acacia in a mortar.

In the case of other un ground or unpowdered drugs, prepare

about 10 Gm. of the official sample by cutting, or shredding, so
that the parts are about 3 mm. in thickness. Seeds or fruits
smaller than 3 mm. should be craeked. High-speed mills, since
they generate considerable heat, should not be used for prepuring
the sample, and care should be taken that no appreciable amount
. of moisture is lost during the preparation and that the portion
taken is representative of the official sample.
The sample should be ground in order to expose as large a
surface of the drug as possible so that the moisture may be driven
off readily.
2. Accurately weigh about 10 Gm. of the drug as prepared in t1 tared
evaporating dish. Dry at a temperature of 100C. for 5 hr., and weigh.
Continue the drying and weighing at 1 hr. interv::LIs until the loss is not
more than 0.25 per cent in 1 hr.'s drying.

The moisture and any other volatile constituents of acaeia are

driven off.
Calculate the per cent moisture contained in the sample of
acacia used.
Questions and Problems
1. Why is 1,he loss in weight of a drug dried at 100C. not alwaYH a true
measure of its moisture content?
2. If an air-dried drug containing 10 pel' cent moisture yielded 2.5 pCI'
cent ash calculated on the basis of the air-dried drug, what per cent ash
would the moisture-free dl11g contain?

Exercise 96
Object.-Determination of the Moisture Content of Digitalis .
by the Toluene Distillation Method.
Materials :Required.-50 Gm, of digitalis leaf.
Toluene Moisture Apparatus,-Use a 500 ce. flask preferably of Pyrex
glass, a straight tube Liebig condenser of about 500 mm. length, and a
332 . QUANTiTATIVE PHARMACEUTICAL CHEMISTRY

moisture-tube receiver calibrated to 0.1 cc. of the type illustrated (Fig. 59).
Olean the condenser and moisture tube with cleaning mixture (see page 5),
rinse with distilled water and then with alcohol, and dry them in an oven at
1000.
Procedure.-l. "Place in the flask an accurately weighed amount of the
drug to be tested, which it is estimated will yield from 2 to 4 ce. of water.
If the drug is likely to cause bumping, add enough dry sanrl to cover the
bottom of the flask. Add sufficient toluene to cover the drug
completely, usually about 75 cc., and connect the apparatus
. as illustmted. Fill the receiving tube with toluene by pouring
it through the top of the condenser. Heat the toltlene in the
flask until it boils, and distil lilowly, about 2 drops per second,
until most of the water. has passed over; then increase the
ra~e of distillation to about 4 drops per second."

Sufficient drug must be taken to yield from 2 to 4

cc. of water because smaller amounts would be
difficult to determiI}e without a relatively large per
cent error: Thus an error of 0.05 cc. in the cali-
bration or reading of the moisture tube would cause
an errol' of 5 per eent if only 1 cc. of ,vater is
obtained and of 1.25 per cent if 4 ce. of water is
obtained. At the boiling point of toluene, 110 to
FIG. 59.- 111 e., the moisture in the drug is volatilized.
Assombled The moisture distils with the toluene, and as the
apparatus for
moisture de- vapors condense and drop into the moisture tube,
termination the water which is immiscible with and heavier than
by toluene
distillution. toluene separates and collects at the bottom of the
moisture tube.
2. "When the water is apparently an over, wash down the condenser
by pouring toluene in at.the top, continuing the distillation a short time to
ascertain whether any more water will distil, and if it does, repeat the wash-
ing of the condenser with toluene. If any water remains in the condenser,
remove it by bmshing it down into the tube receiver with a tube brush
attached to a eopper wire and saturated with toluene, at the same time
washing the condenser with toluene. Allow the receiving tube to stand
until cooled to room temperature and if any drops of water still adhere to
the sides of t.he tube they can be forced down by a rubber band wrapped
around a copper wire. Finally rmld the volume of water and calculate to
determine the percentage which was present in the drug."

Practically all the moisture distils over during the first 4 or

5 min. of distillation. The condenser tube is washed down with
 ABU AND MOISTURE DETERMINA.7'IONS 333
TABLE XLVII.-OFFICIAI, SUBSTANCES WITH MOISTURE LU!ITS

Tem- Tem-
Moisture 1toisture
p.'r- per ..
Substance limit, Substance limit.
attlre, ntllf(1,
pel' cent ppr ~cnt
c. c.

U.S.P. U.S.P.
Acacia .............. . 15 Phenolsulfonphthalein. 110
Acid acetyltannic. . . . . 100 Potassium carbonate .. 180 15
Acid, tannic.. . . . . . . . . 100 12 Potassium citrate .... . 150 3 to 6
Acriflavine. . . . . . . . . . . 100 7 PotasHium iodide ..... . 100 1.5
ACT i II a vi ne, hydro- Potassium and sodiutn
chloride ............ R,SO, 7 tartmte ........... . 150 21 to 26
Agar ............... In Quinidine sulfute: .... . 120 5
Albumin tannate...... 100 6 Quinine., ........ , .. , 100 15
Aloe ................ . 10 Quinine bi"ulfate ..... . 100 24
Alum, exsiaca ted. . . . . . 200 10 Quinine dihy<irouhlo-
Barbital, suluble.. .... 100 ride .............. . 100 3
Caffeine. . .... . . . . ... . . 80 Quinine ethyl carbon-
Caffeine, citra.ted.. .. . . 80 5 ate . ..... , , . , ..... . H,SO. 2
Caffeine with sodium Quinine sulfate ....... . 100 16.2
benzoate....... .. . . 80 5 Scopolamine hydrobro-
Calcium iodobehenate. 100 2 nlide .. , ... , ...... ,. 100 13
Calcium lactate. . . . . . . 120 25 to 30 Soap. " " " " " " ' " 110 36
C"ntharides. . . . . . . . . . 100 10 Soap, powdered . . , , .. . 110 10
Cnsein, R..... . . . . . . . 100 10 Soap, soft ........... . 110 52
Codeine...... .. .. . . .. 80 6 Sodium acetate . ..... . 120 3ll to 41
Codeine sulfate. . . . . . . 100 12 Sodium biphosphate .. . 100 10 to 15
Dextrose.. .. . . . . . . . . . 105 10 Sodium carbonate.
Digi talis ............ . 8 monohydrat.ed ..... . no 10 to 15
Digitalis, powdered ... . 5 Sodium citrate ....... . 1ilD 10 to 13
Emetine hydrochloride 100 8 to 10 Sodium iodide ....... . 12fl 7
Ephedrine hydrochlo- Sodium phosphate ... . 110 43 to 50
ride. . . . . . . . . . . . . .. R,SO. 2 Sodium phosph",te, ex-
Ephedrine sulfate. . . .. R,SO. 2 siccated ........... . 110 5
Ergot ............... . 8 Sotlium Btearate ...... . 110 5
Ethylhydrocuprei ne Sodium sullate ....... . 120 51 to 57
hydrochloride. . . . .. R,SO. Sodium thiosulfate ... . 100 32 to 37
Ethylmorphine hydro- Starch .............. . 14
chloride. . . . . . . . . . . . 100 10 Strychnine sulfate .... 100 11.5
Fluorescin, soluble. . . . 105 5 Theobromine with so-
Gentian ...... ; ...... . 10 dium sulicyllltc .... . 110 10
Glucose.......... ... . 00 21 Theophylline ........ . 100 9.5
Histamine phosphate. . 100 1 Theuphylline with
Imlo[orm. . . . . . . . . . . .. 1I,SO. 1 ethylene din.mine. ; .. IhSO. 4.5
Magnesium oxide .... . Ignite 10 Thyroid ............. . 100 6
Magnesium sulfate ... . Ignite 45 to 52 Tryparsamide ....... . 110 2.5 to 3.5
Merbnphen .......... . 100 2 Wool fut ............ . 100 0.5
Methylthionine chlo- Wool fat, hydrous .... . 100 25 to 30
ride .............. . 110 16 N.F.
Morphine Bulfate ..... . 130 12 Acid, gallic .......... . 100 12
PhenacD.ine hydrochlo- Aluminum sulfate .... . 200 45 to 49
ride .............. 105 7 Ammonium hypophos-
PhenobarbitaL. :: .... 140 7 phite ............. . H,SO, 3
334 QUANTI'l'ATIVE PllARMACEU'l'ICAL CHEMISTRY

TABLE XLVII.-OFFICIAL SUBSTANCES WITH MOISTURE LIMITS

(Continued)

Tom- TelTl~
Moisture Moisture
per- per-
Substance limit, Substance limit,
n.ture, aturc,
per cent
C.
per cent nco

N.F. N.F.
Ammonium iodide .... 110 6 l\1ercuric oxide, red . .. H2S0, 2
Ammonium valerate, Methyl-rosaniline ..... 110 7.5
Hcicl .... ......... .. H2SO4 2 Morphine hydrochlo
Arecoline hydrobro- ride ........... : ... 100 15
nIi(ic .. , .......... . TI2S0. 1 Ovarian residue . ... , .. 6
Brudne sulfa.te .... , .. 100 13 Ovary ............... 6
Calcium glycerophos- Papaverine hydrochlo-
phate .............. 130 10 ride ............... H,SO,
Calcium hypophos- Pilocarpine hydrochlo-
phite .............. H 2SO, 3 ride ... ........... , TI2S0, 3
Calcium phosphate .... 200 4 Pituitnry, anterior ..1 6
Carmine ............. 100 25 Pituitary, whole ...... 6
Charcoal, purified ani- Potassium chloride .... 100
mal. ............... 100 )2 Potassium guaiacolsul-
Cholesterol, R ........ 100 2 fonate ............. H,SO. 2
Cinchonidine sulfate ... 100 12 Potassium hypophos-
Cinchonine fJulfnte .... 100 5 phitp. .. _ ........... H2S0. 5
Cinchophen, ...... ,. 100 2 Potassium thior.yanate 110 5
Corpus luteum ....... . 0 Quinine hydrobromidc. 100 5
Cotarnine chloride .... H2S0. I Quinine hydrochloride, 110 10
Ethyl carbamate ... , .. H,SO., 3 Quinine salicylate ..... 100 5
Ferrio glycerophos- Resin of ipomoea ..... 100 4
phatc .... , ......... 110 5 Resin of jalap ........ 100 4
Ferric hypophoaphite .. H2S0, 3 Saba!. ............... 10 to 15
Hydrastine hydrochlo- Salicin._ ......... .... H2S0. 2
ride ............... n,so. 2 Sodium arsenate .. , , , . 150 3
J,jthium henzoate ..... 100 3 Sodium hypophoBPhite. H,SO, 3
Lithium cltrhonate .... 100 2 Radium sulfate ........ 100 3
Lithium oi tra tc ....... 150 26 Sodium thi{lCYllllute ... 110 5
J.ithium salicylate ..... 100 5 Sparteine sulfate ...... 100 22
Manganese glyocro- Strontium bromide .... 200 32
phosphlLte.......... 110 10 Strontium sulicyll,te ... H,SO. a
Manganese hypophos- Sulfonmethane ........ H2S0.
phite ......... , .... H,SO. 2 Suprarenal. .......... 6
Mercuric iodide, red ... 120 Zino iodide ........... H,SO, 5

R. = reagent. H2S0, = dried Over sulfuric acid.

toluene to remove water which may adhere to its inner wall.

If the condenser is cleaned as directed, little difficulty will be
experienced with water adhering to the wall of the tube: Some
drugs hold their moisture quite tenaciously. Consequently,
it is always well to continue distillation for 2 or 3 min. after
, '
 ASH AND MOISTURE DETERMINATIONS 335

it is apparent from the test that practically all of the moisture

has distilled. Droplets adhering to the wall of a properly
cleaned moisture tube can be dislodged by rotating the tube
between the fingers or by tapping Lhe tube gently on wooel or some
other base. Volatile organic substances, such as volatile oil~
and amines, remain dissolved in the toluene and so are not
calculated as moisture. Since the result is read .directly in
cubic centimeters, it is apparent that the per cent moisture would
be 2 X number of cubic centimeters of water when a 50 Gm.
sample is used.
Questions and Problems
1. Consult Table XL VII and list five vegetable drugs, three glandu-
lar products aud five chemical compouuds the moiHture contents of which
might be determined hy the toluene distillation method.
2. Less than 1 part of wlLter is soluble in 10,000 parts of toluene at 25C.
Would the water dissolved in the toluene when moisture is determined in
a drug by the toluene distillation method be appreciable if 100 cc. of toluene
is employed?

Certain other substances have definite limitations on the

amounts of volatile matter which they will lose when heated,
e.g., when ignited, the loss in weight of prepared calamine =
not more than 1 pel' cent, of kaolin = not more than 15 percent,
of purified siliceous earth = not more than 10 per cent, of
purified talc = not more than 5 per cent, of lead monoxide =
not more than 4 per cent, and ichthammol dried to constant
weight at 100 0 e. loses not more than 50 per cent of its weight.
The loss in weight in these cases cannot be considered as moisture
only, since adsorbed gases as well as other volatile matter are
driven off.
 CHAPTER XXI
EXTRACTIVE AND CRUDE FIBER CONTENT
The amount of extractive which a drug yields to a given solvent
is often an approximate measure of the amount of a certain
constituent or group of related constituents which the drug
contains. In cases where this is the object sought, it is important
that a solvent be used which will not dissolve appreciable quanti-
ties of other substances than those sought in the extraction.
In some cases, the amount of a drug Roluble in a given solvent is
an index of its purity; e.g., acacia must yield not more than 1 per
cent of water-insoluble residue to meet the official requirement.
For efficient and complete extraction, the solvent should be
divided into a number of relatively small portions and used
successively. Each portion should be brought into intimate
contact with all parts of the sample and then be reluoved before
the addition of the next portion. The use of fresh portions of
the solvent which are not saturated with extractive favors the
rapid and thorough removal of the solute with the minimum
quantity of solvent.
The complete removal of extractive matter from a drug by
this method, howevel', requires a large quantity of solvent in most
cases, and most volatile solvents are expensive. Consequently,
it is convenient and economical to be able to use the same portion
of solvent repeatedly. To do so necessitates the separation of
the solvent and solute after the extraction, the separated solvent
being employed in a second extraction. This is readily accom-
plished by means of a special form of extraction apparatus of
which the Soxhlet is a good example.
The Soxhlet (Fig. 60) is the apparatus generally used for
extraction with volatile solvents where small quantities of drug
are extracted as in quantitative work. The solvent is placed in
a weighed flask A, which is heated over an electric hot plate or
over' a water bath in the absence of a gas flame when the solvent
is highly inflammable. The weighed sample of drug is placed
836
 EX1'RAC1'IVE AND CRUDE FIBER CONTENT 337

in a porous paper extraction thimble, which is then fitted into

the extraction tube B. A reflux condenser C, is attached to the
top of the ~xtraction tuhe. The solvent vapors pass upward
from the flask A through the side tube D of the extractor to the
condenser. The condensate drips into the thimble
until a sufficient amount has accumulated to raise
the level of liquid in the extraction tube to the top
of the siphon tube E. The solution which has
been standing in contact with the drug is then
discharged into the flask where the solvent is
vaporized again. The solvent passes through this
cycle repeatedly, and the solute is collected in the
flask. The heat should be so adjust.ed t.hat the
solvent passes around the cycle about once every D
10 min. The top of the t.himble should be above E

the level of the top of the siphon side tube. The

solvent must not be volatilized more rapidly than
it will filter through the porous t.himble, or the
latter will overflow and carry insoluble matter over
mechanically. After the extraction is complete, FIG. 60.-
Soxhlet cJCtrac
the flask is removed, the solvent is driven off until tion applll:atus.
the flask and its contents are of constant weight,
and the percentage of ext.ractive in the drug is calculated from
the weight of the residue.
In some cases, it is advantageous to determine the insoluble
residue after extraction. The extraction thimble should be
dried and weighed accurately before placing the sample in it,
After the extraction is complete, it is dried with the residue and
weighed accurately again. Caution: Do not place thimbles
and residues saturated with ether or other inflammable or
explosive solvents in an electric oven until they have been.
air-dried.
Volatile and Non-volatile Ether-soluble Extractive.-The
determination of the total ether-soluble volatile constituents
is applied to those drugs which contain volatile oils, while the
determination of the non-volatile ether-soluble constituents is
applied to those drugs the active constituents of which are asso-
ciated with :volatile matter. In either case, the determination is
only approximate.
338 QUANTITATIVE PHARMACEU'l'ICAL CHEMISTRY

Exercise 97

Object.-To Determine the Volatile and Non-volatile Ether-

soluble Content of Clove.
Materials Required.-2 Gm. of clove.
50 to 100 ce. of !Lbsolutc ether.
Soxhlct apparatus.
Procedure.-l. "Extract completely 2 GIll. of thc prepared drug, dried
over sulfuric acid for not less than twelve hours, by subjecting it, during
twenty hoUl's, to the action of absolute ether in a continuous extraction
apparatus."

The drug is dried over sulfuric acid in a dessicator to remove

the absorbed moisture. The extraction is continued for 20 hr.
to insure complete removal of the volatile oil and other ether-
soluble constituents from the ground clove. Absolute ether
is used, since ordinary ether, which contains small,' va:r:iable
quantities of water, dissolves some tannin, sugar, etc.
2. "Transfer the ethereal solution to a tnred pOl'cehtin dish ILnd allow it
to evaporate spontaneously. Thon dry it over sulfurio acid dluing eighteen

TAllLE XLVIII.-OFFICIAL SUBSTANCES WITH ETHER-SOLUBLE EXTRAC'l'rvE

Volatile, Non-volatile, Total ether-soluble

Substance
per cent per ccnt extractive, per cent

U.s.P.
Capsicum ............... . 12
Cinnamon ......... ' .... . 2
Clove ............. ' .... . 15
Ginger ......... : .. , .... . 4.5
Ginger, fluidextract of ... . 20 4.5W/V
Linseed ............ ' .... . 30
Myristica ............... . 25
N.F.
Cacao, prepared ......... . 22
Coriander .............. . 0.5
Cubebs ................. . 10
I{amaJa ................ . 70
Lupulin ................ . 60
Mastic ................. . 97
Salvia .................. . 1.0
Sandalwood, white ....... . 3.5
 EXTRACTIVE AND CRUDE FIBER CONTENT 339
hours and weigh the total ether extract. Now heat the extract gradually
up to noc. until the weight becomes constant; the loss in. weight during
the heating represents the volatile portion of the extract."

Since ether boils at 35C., it evaporates rapidly at ordinary

temperatures without carrying off more than negligible amounts
of the volatile extractive. The residue, dried for 18 hr. over
sulfuric acid to get rid of the small amounts of water extracted
by the ether, represents the ether-soluble extractive of the clove.
This residue consists chiefly of volatile oil, eugenol, and resin.
When the residue is heated at 110C. until of constant weight,
the volatile oil and eugenol are volatilized, leaving resin, coloring
matter, etc., as the non-volatile ether-soluble extractive. The
difference in the weight of the .. total extractive' and the non-
volatile residue is the volatile ether-soluble extract of the clove.
Alcohol-soluble Extractive.-Alcohol is a good solvent for
resinous matter. Consequently, the determination of the
alcohol-soluble extractive is most frequently employed to
determine approximately the amount of resin in those drugs in
which the resinous matter is the important constituent. In'
some cases, the determination of the alcohol-soluble extractive
is considered officially as an assay process, e.g., asafoetida and
kino, and in other cases, the extractive upon further purification
is employed in the assay, e.g., jalap.
Exercise 98
Object.-Assay of Benzoin.
Materials Required.-2 Gm. of benzoin.
100 cc. of alcohol.
Procedure.~1. HPlace about 2 Gm. of Benzoin, accurately weighed, in a
tared extraction thimble and about 0.1 Gm. of sodium hydroxide in the
receiving flask of the extraction apparatus and extract with alcohol in It
Soxhlet apparatus or other suitable extraction apparatus for five hours, or
until completely extracted."
The alcohol dissolves the resins, benzoic acid, cinnamic acid,
styrene, benzaldehyde, vanillin, esters, etc., contained in the
balsamic resin. The NaOH in the receiving flask forms non-
volatile salts of the volatile acids.
2. "Dry the insoluble residue at 100C. for thirty minutes and weigh.
Dete!'lIline the amount of moisture in the drug by the toluenp. distillation,
,340 QUAN1'ITA'l'IVE PHARMACEUTICAL CHEMIS'1'RY

method, page 331, calculate the weight. of moisture in the Benzoin and sub-
tract this weight of rnoistnre from the original weight of the Benzoin taken
for 'the assay, The difference between this result and the weight of th~
residue determined above represents the alcohol-soluble extractive."

The residue should be dried and weighed in the extraction

thimble. When the extraction is complete, the thimble should
be removed from the apparatus and allowed to drain and dry in
air for about 5 min. to remove most of the alcohol. The residue
and thimble can be dried conveniently at 100oe. in an electric
oven. Since moisture remaining in the marc as w('U as the alcohol
is volatilized at 100C" it is necessary to correct for the loss in
moisture by a moisture determination.
Calculate the per cent of alcohol-soluble extractive assuming
thn,t thc benzoin contained 3.20 per cent moisture.
The diluted alcohol-soluble extractive is determined in the
same way, using diluted alcohol as the solvent,
TABLE XLIX,-OFFICIAL SUBS'rANeES WITH ALCOHOIrSOI"URLE EX'l'RACTIVE
LIMITS

Alcohol used, Extractive,

Substance
per cent pet cent

U.S.P.
Asafoetida ..... , , . , .... . 94 50
Benzoin, Sumatra, . , .... . 94 75
Benzoin, Siam. , , ..... , .. 94 00
Bismuth and pot.assium
tartrate ..... , .. , .. , .. , 94 (n.m.t.) 0.5
Bismuth subgallate. , , , , , , 94 (n.m.t.) 0.5
Kino, , .. , . , ....... , . , . , 94 60
Myrrh, .. , , ... , ...... , .. ' 94 30
Rhubarb ....... , ... ,.,., 40 30
N.F.
Chionanthus ......... , .. 73 25
Gambogc, ............. , 94 65
Gambit .. , .... , ...... , . , 94 60
Guaiac .. , . , ... , ..... , , , 94 85
Manna, , , , .......... , .. 94 75
Mastic .. , . , , . , , .. , , .. , .. 95 80
Poplar bud, ........ , ... . 05 40
Vanilla .. , .... , ........ . 49 12
(n.m.t.) = not more than.
 EXTRACTIVE AND CRUDE FIBER CON'l'ENT :341

Water-soluble Extractive and Water-insoluble Residue.-

The determination of the water-soluble extractive content is
applied to those drugs one or more of the most important con-
stituents of which are soluble in water, e.g., aloe, gambir, and
kino. The determination is performed in the same wayaH the
determination of the alcohol-soluble extractive. The deter-
mination .of the amount of. water-insoluble residue is used as a
test for the purity of some substances. ThuH, it is required
that acacia yield not more than 1 per cent, aloin yield not more
than 1.5 per cent, acetyitannic acid yield not more than 6 per
cent, and acriflavine yield not more than 0.5 per cent of water-
inHoluble residue.
Exercise 99
Object.-Assay of Aloe.
Materials Required.-2 Gm. of aloe.
Procedure.-Proceed as directed. in the assay of benzoin, Exercise 97,
llsing distilled water instead of alcohol as the solvent.
Barbaloin, isobarbaloin, and other active constituents of a
glycosidalnature dissolve in the water, leaving resinous matter,
etc., undissolved.
The U.S.P. requires that aloe contain not less than 50 per cent
of water-soluble extractive. Calculate the percentage of water-
soluble extractive contained in the sample examined and compare
the result with the official requirement.
TABLE L.-OFl!'ICIAL SUBS'l'ANCES WITH WATER-SOLUBLE EXTR.A.C'l'lVE LIMITS
Water-tioillble
Extractive,
Substance Per Cent
U.S.P.
Aloe ............................................. . 50
Gentian .............................. - .. , ....... , . 30
Ginger ................................... , ...... . 12
Kino., ........................ -., ............... . 80.
N.F.
Galnbir ........................................... 70

Purified Petroleum Benzin Extractive.-Purified petroleum

benzin is a good solvent for fats and fatty oils. The determina-
tion of the amount of extractive with this solvent is performed
. as follows:
342 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

II Extract completely about 2 Gm, of the prepared drug,

accurately weighed, by subjecting it during twenty hours to the

action of purified petroleum benzin in a continuous extraction
apparatus, Transfer the benzin solution to a tared porcelain
dish and allow it to evaporate spontaneously. Then dry it
over sulfuric acid for eighteen hours and weigh. Calculate the
percentage of anhydrous extractive from the weight of drug
taken,"
Colocynth is the only official drug for which a petroleum
benzin extractive limitis given. In this case the determination
serves as a check on the amount of seeds contained in the
colocynth pulp, since the seeds contain a large amount of fatty
matter.
Crude Fiber.-The crude fiber content of a drug is the residue,
consisting chiefly of cellulose, which remains undissolved after
successive treatment with boiling acid and alkali. The deter-
mination of crude fiber is of considerable importance in the
examination of certain drugs and particularly of spices, since the
commonly used adulterants consist of waste or refuse material
derived from the drugs or spices themselves or from other food
products. Frequently, this material is the outer cellular layer
or protective coating which contains a larger proportion of
lignified tissues and, consequently, more crude fiber than the
part official.
Exercise 100

Object.-To Determine the Crude Fiber Content of Clove.

Materials Required.-2 Gm. of clove in fine powder.
200 ce. of 1.25 per cent H 2SO .
200 cc. of 1.25 per cent NaOH.
Procedure.-l. "Exhaust a weighed quantity of the prepared drug
representing about 2 Gm. of the drug, with ethel' or use the residue from the
determination of the volatile ether-soluble extractive."

The drug should be powdered as finely as possible so that the

acid and alkali will penetrate the innermost tissues and thus
insure complete removal of soluble material. It is exhausted
first with ether to remove fats and waxes, which, being immiscible
with aqueous solutions, tend to prevent the penetration of the
acid and alkali solution into the drug particles.
 EXTRACTIVE AND CRUDE FIBER CONTENT 343

2. "Add 200 cc. of boiling sulfuric acid solution, adjusted to exactly

1.25 per cent by titration, to the ether-exhausted drug, in a 500 ce. flask,
and connect the flask with a reflux condenser, the tube of which passes only
a short distance below the rubber stopper, into the flask. Heat the mixturc
to boiling and continue the boiling exactly thirty minutes. Then filter
the mixture through a linen or hardened-paper filter and wash the residue
on the filter with boiling distilled water until the washings are no longer
acid."

By diluting exactly 51 cc. of normal sulfuric acid or an equiva~

lent amount of a standard acid of known normality to 200 cc.
at 25C., 200 cc. of 1.25 per cent sulfuric acid may conveniently
be prepared. The prepared acid solution should be measured
and heated to boiling before it is added to the drug. When
the acid solution is added to the powdered, ether-exhausted
drug contained in a cold flask, the solution is cooled below its
boiling point. The mixture should be heated rapidly to bring
it to the boiling point again, and thereafter the flame should be
cut down to the minimum required to maintain slow, steady
boiling. Since the amount of material dissolved by the acid
solution is partly dependent upon the length of time that the
latter is heated in contact with the sample, it is important that
the mixture should be boiled exactly 30 min., the time period
being taken from the time the mixture starts to boil and not
from the time the hot acid solution is added. The flask should
be rotated gently from time to time while the mixture is being
boiled to bring particles of the drug which adhere to the walls
of the flask into contact with the acid. A current of air, passed
into and beneath the surface of the mixture, through a capillary
tube, will frequently prevent excessive frothing. The acid-
insoluble residue is collected on a filter and washed until free of
acid, since the latter, if present, would neutralize some of and,
consequ:ntly, reduce the concentration of the alkali used in the
next step of the procedure.
3. "Rinse the residue back into the flask with 200 cc. of boiling sodium
hydroxide solution, adjusted to exactly 1.25 per cent by titration and free
from sodium carbonate. Again hcat the mixture to boiling and continue
the boiling exactly 30 min. under the reflux condenser as described under
the treatment with acid, then rapidly filter through a tared filter, wash the
residue with boiling distilled water until the last washing is neutral, dry it at
nOc. l,lntil of constant weight, and note the weight."
344 (jUllNTl'l'A.TIVE PHARMACEUTICAL CIIEMIS'1'RY

The alkali solution may be prepared conveniently by diluting

70 cc. of recently standardized, carbonate-free, normal sodium
hydroxide or an equivalent quantity of standard alkali solution
of known normality to exactly 200 cc. The residue from the
acid treatment in step 2 should be boiled with alkali in a manner
duplicating as nearly as possible that used in the boiling with acid.
The residue is then collected on a filter, preferably in a Gooch
crucible, washed free of alkali, dried to drive off moisture, and
weighed accurately .. The dried residue consists chiefly of cellu-
lose and small amounts of hemicelluloses, pentoses, inorganic
matter, etc.
4. Incinerate the dried residue, and weigh the ash.

The fibrous matter should be incinerated cautiously at first and

then at a low red heat until the carbon is completely burned off
as indicated by the formation of a white ash. The cellulose, etc.,
is burned when the fiber is incinerated, leaving the inorganic
matter. The weight of the ash obtained subtracted from the
weight of the residue found in step 3 gives the weight of the crude
fiber.
The Pharmacopoeia requires that clove contain not more than
10 pel' cent of crude fiber. Calculate the percentage of crude
fiber contained in the sample examined and compare the results
with the official requirements.
Salvia is required to yield not more than 25 per cent of crude
fiber. The ether-insoluble residue of prepared cacao of the
National Formulary is required to contain not more than 7 per
cent of .crude fiber.
Questions and Problems
1. What is meant by the term crude fiber?
.2. Why, should the sample used be finely ground and exhm~:~ted with
ether before it is boiled with acid and alkali?
3. Why should the boiling with 1.25 per cent acid and alkuli be continued
for exuctly 30 IHin. ?
4. What substances remain undissolvlld ,by the [lcid and alkali?
 CHAPTER XXII

CONSTANTS OF FATS, FATTY OILS, WAXES, BALSAMS,

RESINS, ETC.
The methods of analysis of fatty substances, waxes, resins, etc.,
usually consist of the determination of a number of physical
and chemical properties or values commonly known as constants,
although they are constant only within certain limits, These
constants, when taken in conjunction with the color, odor, taste,
and special identity tests for the given substance and for common
adulterants, are the basis upon which the purity and quality of
these substances are judged. Although the following determina-
tions are not required of all of the official substances which fall
under the above caption, they are generally used in the examina-
tion of such substances,. .'
The specific gravity, solubility, melting point, congealing point,
refractive index, and optical activity determinations are per-
formed according to the methods which haye been discussed
previously (pages 203 to 256).
Preparation of the Sample of Fats and Oils.----''' If a sample
of oil shows turbidity due to separated stearin, Warm the con-
tainer in a bath of water at 50C. until the tUl'bidity has dis-
appeared and the oil is clear. Thoroughly mix the clarified
oil before weighing the samples. If the turbidity is due to other
than separated stearin, i.e., if the oil does not become clear on
warming, filter it through dry filter paper in a funncl contained
in a hot water jacket. Weigh out at one time as many portions
as are needed for the various determinations, using preferably a
bottle having a pipette dropper, or a weighing burette. Keep
the sample molten, if solid at room temperature."
Acid Value.-The acid value, also known as the acid number
and as the acidity index, is defined as the number of milligrams
of potassium hydroxide necessary to neutralize the free acids in
1 Gm. of oil, fat, wax, resin, balsam, or similar organic substance.
345
346 QUANTITATIY.E PHARMACEUTICAL CHEMISTRY

of complex composi#on. In other words, it gives the amount of

potassium hydroxide, expressed in tenths of 1 per cent, required
to neutralize the free acids in a substance. The acidity may also
be expressed as the number of cubic centimeters of 0.1 N NaOH
required to neutralize the free aeid in 10 Gm. of substance. This
value is determined by titl'tl-ting a weighed sample of the sub-
stance, contained in an alcoholic or in an alcohol-ether solution,
with standard alkali solution, using phenolphthalein as the
indicator. For the titration, 0.5 N, 0.1 N, or 0.02 N alkali may
be employed, but a solution of 0.1 N concentration is most
suitable in the majority of the official determinations. In the
case of substances like the balsams and resins, a mixed solvent
such as alcohol and ether may be used advantageously, since the
coloring matter which interferes with the observation of the end
point is dissolved in the ethereal layer when the alkali is added.
Solid fats and waxes are usually melted on a water bath and
titrated while hot.
The presence of free acids in the oils, fats, and waxes is due
chiefly to the hydrolysis of the esters composing them and
is caused by chemical treatment, by bacterial action, or by
the catalytic action of light and heat. As a rule, fresh or recently
prepared fatty substances contain little or no free acids. Upon
aging, the acid value increases slowly at first and more rapidly
later, especially if the substance is not well protected from
the simultaneous action of light and air. The acid values
of the official fatty substances are quite variable; consequently,
the official standards fix maximum limits, in most cases, which
if exceeded indicate that the substances have undergone hydro-
lytic decomposition in their. preparation, purification, or during
the period of storage. High acid values are not necessarily an
indication of rancidity, since the latter is a result of the action of
the air, or possibly bacteria, on the liberated fatty acids.
, A minimum acid value or a minimum and a maximum acid
value is generally given in the official standards for substances
which are balsamic or resinous in character. The value of these
substances is usually based, in part at least, upon their content
of free acids; e.g., the acid value of tolu is required to be not
less than 112 and not more than 168. Since a good grade of
tolu usually contains from 12 to 15 pel' cent of cinnamic and
 CONSTANTS OF FATS, FA'l'TY OILS, WAXES, ETC. 347

benzoic acid in the free state, an acid value of less than 112 would
indicate that the acid content of the balsam was low and that it
was of inferior quality or adulterated. On the other hand, an
acid value greater than 168 would indicate adulteration with
some substance having high acid value, such as certain resins.
Exercise 101
Object.-To Determine the Acid Value of Rosin.
Materials Required.-About 1 Gm. of rosin.
0.5 N potassium hydroxide.
About 50 {\c. of alcohol.
Procedure.-l, Pulverize the rosin in a mortar and dissolve about 1 Gm.
of the powder, accurately weighed in from 40 to 50 cc. of neutral alcohol.

The rosin is powdered to facilitate its solution. Oommercial

alcohol is frequently acid in reaction and it should always be
tested before the sample is dissolved. This test may conven-
iently be carried out in this case by adding 1 cc. of phenolphthalein
indicator solution to the alcohol before the rosin is dissolved.
If the alcohol is found to be acidic, it should be rendered neutral
to phenolphthalein by the addition of the standard alkali
solution.
In the titration of substances which form colored solutions,
the end point may frequently be made observable by dilution
with alcohol to it volume of from 100 to 200 cc.
2. Add 1 ce. of phenolphthalein indicator solution (if it has not already
been added ill testing the neutrality of the alcohol) and titrate the solution
with 0.1 N NaOH, agitating the mixture continuously.

In the titration of fatty substances, it is necessary to shake the

mixture thoroughly after the addition of each portion of alkali to
secure complete extraction of the fatty acids from the immiscible
oily layer. This precaution is not of great importance in the
titration of rosin, however) since rosil1 is soluble in alcohol and
also in alkalies.
The alkali reacts with the dibasic abietic acid formed from the
abietic acid anhydride and with the other acids contained
in the rosin with the formation of so-called rosin soaps and water.
It has been suggested that the acid value of rosin can be more
accurately determined in alcoholic solution, using alcoholic
348 (.!U.IN1'I'J'.U'1V]i} PJURMJl(!E[JTICA.L CHEMISTRY

KOH in the titration, since the presence of water in the above

determination eauses dissociation of the rosin soap with conse-
quent low results.
The calculation of the acid value is simple; e.g., if a 1 Gm.
sample of rosin required 30 ce. of 0.1 N NaOH for titration, the
acid value would be 30 X 5.612 = 168.36, where 5.6]2 is the
1
number of milligrams of KOH equivalent to 1 cc. of 0.1 N NaOH
solution.
Questions and Problems
1. Define the term acid value.
2. Why must the alcohol or ether used as a solvent for fats, resins, etc.,
in acid v!1lue determinations be neutral?
3. Look up the U.S.P. te~t for free fatty acids under lard and explain
why this test may serve as a criterion of the purity of this fat.
4. Why do the official standards set up maximuIll acid values for most of
the official fats and oils a~d minimum valnes for most of the resinous
substances?
TABLE LI.-AcID VALUE LIMITS OF OFFICLU, SUBSTANCES

Amount Official
Alkali
Substance used, requirement,
used
Gm. acid value

U.S.P.
Acid, oleic ................ 10 0.1 N NaOH 188 to 200
Copaiba .................. 2 0.5 N KOH 28 to 95
Peruvian babarn .......... 1 0.5 N NaOH 56 to 84
Rosin .... " .............. 10 0.1 NN!\OH Not less than 150
Soap, hard (acids) ......... 10 0.1 N NaOH 185 to .205
Soap, soft (acids) .......... 10 0.1 N NaOH 190 to 205
Storax, American .......... 1 0.5 N N!,OH 38 to 85
Storax, Levant ............ 1 0.5 N NaOH ,56 to 85
Tolu balsam ..... " ........ 1 0.5 N KOR "~I!"~ 112 to 168
(alcoholic)
White wax ................ 3 0.5 N KOH .1'7.to 23
1".;. "
(alcoholic)
Yellow wax....... '...... : . ,. 13 0.5 N KOH 18 to 24
,. (alcoholic)
N.F.
Mastic ..... '. : .. ".... '...... 10 .1 N NaOH Not less than 50
Resin of ipomoea'. : ... ' .. : .. 2 .5 N;KOH 8.5 to 18'
.. . .
- . 1 ~ , .. ! '
 CONSTANTS OF' FATS, F'AT1'Y OILS, WAXE'S, ETC. 349
6. If a 2 Gm. sample of cod liver oil requil'ed 4.5 ceo of 0.02 N NaOH in
the titration of the free fatty acids, w(mld the oon eonform to the oflieial
purity -requirement? Whitt would be the acid number of the oil?
6. Calcul!1te the minimuIll and maximum acid values permitted under
the test~ for purity of chaulmoogra oil.
In addition to the listed substances for which definite acid
0

value limits are given in the official Rtandards, there are a number
of substances for which the maximum content of free fatty acids
is fixed by the volume of the standard alkali solution required
in their titration; e.g., 2 Gm. of cod liver oil must require not
more than 1 ce. of 0.1 N NaOH, and 1 Gm. of prepared suet 0

shouid not require more than 0.6 ce. of 0.1 N NaOH to neutralize
the free fatty acids.
Saponification Value.-The saponification value, saponification
number, or Koettsdorfer number, as it is sometimes called from
the originator of the process, is defined as the number of milligmms
of potassillm hydroxide required to neutralize the free acids and
saponify the esters contained in 1 Gm. of oil, fat, wax, or other
substance of similar composition. This value represents the
amount of potassium hydroxide, expressed in tenths of 1 pel' cent,
required to neutralize the total free and combined acids in 1 Gm.
of the substance, or, in other words, it is ten times the percentage
o of potassium hydroxide required to neutralize all of the acids
contained in the sample after saponification. Since the natural
fats and oils consist of mixtures of glyceryl esters of the higher
acids, their saponification values do not differ greatly. The
determination of the saponification value, however, serves to aid
in the detection of the presence of the glycerides of acids con-
taining less than 16 or more than 18 carbon atoms, since the value
of this constant is inversely proportional to the mean molecular
weights of the aeids present. In some cases, it may also indicate
adulteration of the sample with unsaponifiable matter, such as
mineral oil.
Exercise 102
Object.-To Determine the Saponificati~n Value of Cottonseed
Oil. 0

Materials Required.-2 Gm. of cottonseed oil.

50 ee. of 0.5 N alcoholic potassium hydroxide (see page 365).
About 50 cc. 0.5 N hydrochloric,: acid.
350 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

"If the oil has been saturated with carbon dioxide for the purpose of pres-
ervation, it should be exposed in a shallow dish in a vacuum desiccator for
twenty-four hoUl's before the portions are weighed for this determination."
Procedure.-l. Place from 1.5 to 2 Gm. of the SfLmple, accurately weighed,
in a fi!lsk of from 200 to 250 ec. capacity, and add to it exactly 25 cc. of
alcoholic 0.5 N potassiulll hydroxide. Insert into the neck of the flask, by
means of a perforated stoppel', a glass tube from 70 to 80 em. in length and
from 5 to 8 mm. in diltmetel', and heat the fbsk on It water bath for 72 hr.,
frequently rotating the contents.

If the sample is clear, it need not be filtered as a rule. When

the presence of extraneous matter, however, makes it necessary
to filter the oil, the operation may be carried out easily by means
of a suction filter. The sample of oil should be introduced into
the weighed flask by means of a dropper to avoid the possibility
of a portion adhering to the neck of the flask, and the whole
reweighed. The long glass tube serves as an air condenser to
prevent the escape of alcohol vapor. An ordinary water-
jacketed condenser may be used with equally satisfactory results.
The mixture of oil and 0.5 N aleoholic KOH is heated on a water
bath and mixed by imparting a rotatory motion to the flask
from time to time in order to accelerate the saponification.
The alcoholic KOH solutioIl used should not be deeply colored
(see page 366), since the color would interfere with the observa-
tioIl of the end point in the titration of the saponification mixture.
Alcoholic KOH, rather than an aqueous solution, is employed,
because the oils are more soluble in alcohol than in water, and
because the products of saponification are completely s"oluble in
alcohol, whereas when aqueous solutions are used, the unsaponi-
fiable niatter remains insoluble.
When saponification is complete, the mixture should be clear
and free from oil globules. Although the glyoeryl esters present
in fatty oils vary greatly with respect to their acid components,
the character of the saponification reaction may be illustrated
as follows:

CaHs(C17HssC02)a + 3KOH-~CsH5(OH)3 + 3C 17H 33 C0 2 K

glyceryl oleate . glycerin potassium oleate

2. Then add 1 ce. of phenolphthalein T.S. !lnd titrate the excess of

potassium hydroxide with 0.5 N hydrochloric acid.
 OONSTANTS OF FATS, FATTY OILS, WAXES, ETO. 351
The approximate amount of alkali consumed in the saponifica-
tion of the oil is found by subtracting the volume of 0.5 N HCI
used in the residual titration from the volume of 0.5 N alcoholic
ROH originally added. Sulfuric acid should not be substituted
for hydrochloric acid, since potassium sulfate would be precipi-
tated from the alcoholic solution and this would interfere with
the observation of the end point.

3. Make a blank test at the same time, using exactly the same amount
of alcoholic 0.5 N potassium hydroxide. The differellce in the number of
cubic centimeters of 0.5 N hydrochloric acid consumed in the actual test
and the blank, multiplied by 28.06 and divided by the weight of the sampl('
taken, gives the saponification value.

The blank test should be carried out at the same time as the
sample is run, using similar flasks, boiling for the same length of
time and under similar conditions, except that the oil should be
omitted. This is done to p.liminate as far as possible errors from
every source such as those which would be introduced by the
absorption of CO 2 by the alkali or by the alkalinity of the glass.
The saponification value may be calculated as in the following
example: 1.532 Gm. of cottonseed oil saponified with 25 cc. of
0.5 N alcoholic KOH required 11.0 ec. of 0.5 N HCI to back
titrate the excess alkali. In the blank test 21.5 cc. of 0.5 N HCI
were required to titrate the alkali. Therefore a quantity of
. h d d d (21.5 - 11.0) 0.0561
potassmm y rOXl e correspon mg to 2 X

1,000 = 294.5 mg. of KOH was consumed in the saponification,

and the number of milligrams of KOH required to saponify 1
Gm. of the oil is 294.5/1.532 = 192.2 mg. of ROn. The sapon-
ification value of the sample of cottonscfld oil is therefore 192.2.
Since 1 cc. of 0.5 N HCI is equivalent to 1 cc. of 0.5 N KOH
or to 2 ~6;,~00 = 0.02806 Gm. KOH, the saponification value
may be calculated more simply from thc formula:
cc. 0.5 N HCI X 28.06 .. 10.5 X 28.00
f
wt,o samp e
1 = sapomficatlOn value, or 1 532
,
== 192.2 mg. l{OH required to saponify the esters and neutral-
ize the free fatty acids in 1 Gm. of the sample,
352 (JUANTI'l'A'I'IVE PHARMACEUTICAL CHEMISTRY

The saponification values of the official fats, 'waxes, resins, etc.,

are determined in the same way;
Questions and Problems
1. Define the term saponification value.
2. Why should a blank determination be run parallel with t,he determina-
tion of the saponifiea1,ion value of the sample?
3. Consult the following table and enumIJmtIJ several oils which, if used to
nduHerate croton oil, might lower its saponification value.
4. A sample of chaulmoogra oil weighing 1.600 Gm. saponified with 25 cc.
of 0.4 N KOH required 9.0 cc. of 0.5 N HCI to titrate the excess KOH.
In the blank determination 20 CC. of 0.5 N HCl was required to titrate the
alkali. Calculate the saponification value of the sample. Does the value
found correspond with the U.S.P. requirement?
5. Need the alcoholic KOH used in a saponification value determination
l)e of an exact known normality?
6. How is the saponification value of balsam of Peru determined in the
U.S.P.? Does the method correspond with that given in the preceding
exercise? '
TABLE LII.-SAPONIFICNl'ION VALUE LIMITS oro' OFFICIAL SUBSTANCES

Amount Saponifi-
Substance used, cation
Gm. value

U.S.P.
Bals(lm of Peril ................ 3 235 to 238
Castor oil. ., " ................ 1.5 to 2 179 to 185
Ch(lulmoogra oil. .............. 1. 5 to 2 190 to 213
Cod JiveI' oil. .................. 1.5 to 2 180 to 192
Corn oiL ..................... 1.5 to 2 188 to 193
Cottonseed oil ...... : .......... 1.5 to 2 190 to 198
Ethyl ch(lulmoograte ........... 1. 5 to 2 190 to 196
ExpI'essed oil of almond ......... 1.5 to 2 191 to 200
Lard ......................... 1.5 to 2 195 to 203
Linseed oil. ......... ........ . 1.5 to 2 187 to 195 .
Oil of tlleobroma ....... , .... , .. 1. 5 to 2 188 to 195
Olive oil .................. , ... 1.5 to 2 190 to 195
Prepared suet ................. 1.5 to 2 193 to 200
Storax ...................... , . 1.5 to 2 160 to 200
Tolu balsam ................... 1 154 to 220
N.F.
Croton oil .................. : . ~ 1.5 to 2 200to 215
Resin of ipomoea .............. 1.5 to 2 170 to 190
Sesame oil ................ , ... 1 188 to 193
 CONSTAN'l'S OF FATS, FATTY OILS, WAXES, ETC. 353

Ester Number.-The ester number, or ester valne, is defined

as the number of milligmms of potassiwrn hydroxide required to
saponify the esters in 1 Gm. of a fat, oil, wax, bctlsam, re.sin, or
similar' organic s1{,bstance. In those substances which do not
eontain free acids, the ester number is equal to the saponification
value. When free acids are present, however, the ester value
is given by the difference between the acid and saponification
values.
The ester value is of particular importance in the analysis
of beeswax, since it serves, in many cases, to indicate the presence
of adulterants such as paraffin. Considered in conjunction with
the acid value, the ester value may aid in the detection of such
adulterants as rosin and stcaric aeid in the wax.
The United States Pharmacopoeia gives the following direc-
tions for the determination of the ester value:
"Shake from 1.5 to 2 Gm. of the substance, accurately weighed
in a 200 to 250 cc. tared flask, with from 20 to 30 cc. of alcohol,
add 1 cc. of phenolphthalein T.S. and titrate with half-normal
alcoholic potassium hydroxide until the free acid is neutralized.
Add exactly 25 ceo of half-normal alcoholic potaRsiul1l hydroxide
and proeeed as directed under "Saponification Value" beginning
with "Insert in the neck of the flaRk" and omitting the further
addition of phenolphthalein T.S. The difference in the number
of ee. of half-normal hydrochloric: acid consumed in the actual
test and in the blank, llluitiplied by 28.06 and divided by the
weight of the substance taken, gives the ester value."
Questions and Problems
1. Define the term ester number.
2. If a sample of beeswax is found to have an acid number of 20.4 and a
I-laponification value of 89.8, what. would be the ester number of the sample?
8. If a sample of white beeswax has an aeid value of 18.45 and an ester
'''':llue of 74, what would be the saponifination value of the sample?
TABLE LIII.-OFFICIAL SUBSTANCES WITH THEIR REQUIRED ESTER
NUMBERS
Substanne Official Ester Value
U.S.P.
White wax. ' .... . . . . . . . . . . . . . . . . . . . . . . . . ... 72 to 79
Yellow wax .......................... ,..... 72 to 77
N.F.
Resin of ipomoea ............. , , , ........... 160 to 180
354 QUANTITATIVE PHARMAOEUTICAL CHEMISTRY

Unsaponifiable Matter.-The term un8aponifiable matter is

defined in the Pharmacopoeia as the substances present in oils
or fats that are not saponified by alkali hydroxides and are insoluble
in water.
When oils and fats are saponified, there remains a small amount
of residue which may consist of phytosterol in the vegetable oils
and fats, of cholesterol in the animal oils and fats, and in some
cases, of unsaponifiable substances added for the purpose of
adulteration. The normal unsaponifiable matter present in
most oils and fats dissolves in the alcoholic soap solution obtained
upon saponification. When a large amount of unsaponifiable
matter is present, however, it may separate out and float on the
surface of the mixture.
Thc determination of the amount of un saponifiable matter
present in certain oils such as linseed oil and cod liver oil may be
indicative of the quality and pmity of the oil. Thus, the addition
of petroleum oils, a common adulterant, to linseed oil would
greatly increase the unsaponifiable matter content of the latter.
The Pharmacopoeia gives the following procedure for the
determination of the amount of unsaponifiable matter in a fat
or wax:
Weigh 5 Gm. of the oil or fat into a 250 cc. Erlenmeyer flask,
add a solution of 2 Gm. of potassium hydroxide in 40 cc. of
alcohol, and heat under a reflux condenser for 2 hr. keeping the
alcohol gently boiling. Evaporate the alcohol on a water bath,
dissolve the residue in 50 cc. of hot distilled water, and transfer
the solution to a separatory funnel, rinsing the flask with two
25 cc. portions of hot distilled water which are added to the
separator. Cool to room temperature, and extract with two
successive portions of 50 cc. each of ether, adding a few drops of
alcohol to facilitate the separation of the two liquids. Combine
the ether extracts in another separatory funnel, and wash the
ether solution first with 20 cc. of an aqueous solution of sodium
hydroxide (4 in 1,000), then with 20 cc. of an aqueous solution
of sodium hydroxide (8 in 1,000), and finally with 15 cc. portions
of distilled water until the last washing is not reddened by the
addition of 2 drops of phenolphthalein T.S. Transfer the
ethereal solution to a tared beaker, and rinse the separator with
10 cc. of ether, adding the rinsings to the beaker. Evaporate
 CONSTANTS OF FATS, FAT'l'Y OILS, WAXES, E'1'C. 355

the ether just to dryness on a water bath, and dry the residue for
30 min. at 100C. Cool the beaker in a desiccator for 30 min.:
and weigh the residue of unsaponifiable matter.
The Pharmacopoeia requires that both cod liver oil and linseed
oil should not contain more than 1.5 per cent and that corn oil
should contain not more than 2 per cent of unsaponifiable matter.
Iodine Value.-The iodine value, or number, is the number of
grams of iodine absorbed by 100 Gm. of oil, fat, wax, or other 81tb-
stance 'under specified conditions. This value is a quantitative
measure of the proportion of unsaturated fatty acids present,
both free and combined as esters, which have the property of
absorbing iodine.
The determination of the iodine number of fats and oils is
important, since it serves to characterize them and to indicate
whether they are pure or admixtures. The so-called drying oils,
such as linseed oil, and the fish oils, such as cod liver oil, have
very high iodine numbers, usually above 120, since they contain
a large proportion of unsaturated fatty acids; the non-drying oils,
such as olive oil and almond oil, have relatively low iodine
numbers, below 100; and the semidrying oils, such as cottonseed
oil and sesame oil, have intermediate iodine values, that is,
between 100 and 120. In the case of the animal fats, the iodine
number is not very high, usually being less than 90. The deter-
mination of the iodine number, therefore, not only serves as an
aid to the identification of known oils, but it also serves to indi-
cate in a definite manner the class to which an unknown fat or
oil belongs. Furthermore, when the iodine number is considered
in conjunction with the saponification value of a fat or oil, it
serves as a means of detecting adulteration, and frequently it
indicates the nature of the adulterant; e.g., olive oil might be
adulterated with cottonseed oil without changing the saponifica-
tion value appreciably, but the iodine number of the olive oil
would be increased. Again, castor oil might be adulterated
with olive oil without changing the iodine number greatly, but
the saponification value of the castor oil would be increased.
Several methods have been developed for the determination
of the iodine number of fats and oils. ,These methods are
generally designated by the name of their originators, as, for
example, the Hubl, Hanus, and Wijs methods. The method
356 QUANTI'l'ATIVE PIlARlIL4CEU'l'ICAL CIIEMI8'l'RY

given in the Pharmacopoeia is essentially that of Hanus. In thiR

method, the following solutions are required:
1. Iodobrornide Test Solution.-This solution is prepared as
follows:
"Dissolve 13.2 Gm. of reagent iodine in 1,000 co. of glacial acetic acid
with the aid of gentle heat if necessary. Cool the solution to 25C .. and
determine the iodine content in 20 cc. by tit.ration with tenth-normal sodium
thiosulfate, then acid to the remainder of the solution a quantity of bromine
chemically equivalent to that of the iodine present. Prel:lerve in glass-
stoppered bottles, protected from light."

No difficulty will be experienced in dissolving the iodine, as a

rule, if the glacial acetic acid contains less than 0.5 per cent of
water. The concentration of this solution is determined by
titration with 0.1 N sodium thiosulfate (see page 164). Thus,
if 20.5 cc. of the iodine solution requires 20 cc. of 0.1 N sodium
thiosulfate, each cubic centimeter of the iodine solution must
contain :O~5 X 0.012692 = 0.01238 Gm. iodine, and the remain-
ing 980 cc. of iodine test solution would contain 980 X 0.01238 =
12.13 Gm. of iodine. Since 126.92 Gm. of iodine is equivalent to
79.92 Gm. of bromine, the quantity of bromine equivalenf tc
the iodine contained in the solution may be calculated.

X :12.13 ::79.92 :126.92

X = 7.64 Gm. bromine

Since bromine has a specific gravity of about 3.1, 7.64 Gm. of

bromine is equivalent to 7.64/3;1 = 2.5 ce. in round numhers.
Inasmuch as a slight excess of iodine does no harm, the addition
of 2.25 ce. of bromine to the iodine solution would be sufficient
in this case.
The bromine may be drawn into a measuring pipette by means
of a rubber bulb and then added to the cool iodine solution.
(Never draw bromine into a pipette by means of the mouth.)
2. Potassium iodide test solution prepared by clissolv:ng 16.5
Gm. of potassium iodide in sufficientwatel' to make 100 ce.
3. 0.1 N Sodium thio8ulfate sol'ution prepared and standardized
as directed on page 158.
4. Starch test solution prepared as follows:
 CONSTAN'i'S Oft' FltTS, [l'A'l'')'}' OILS, lVl1XE'S, ETC. 357

Triturate 1 Gm. of arrowroot atlnch with 10 ceo of cold distilled waier,

add sufficient boiling distilled water, with constant stirring, to make 200 ce.,
then boil the mixture until a thin translucent fluid is obtained. The solution
must be freshly prepared and filtered, if necessary.

Exercise 103
Object.-To Determine the Iodine Value of Olive Oil.
Materials Required.-About 10m. of olive oil.
20 ceo of chIOloform.
50 cc. of iodobromide test solution.
60 cc. of potassium iodide test solution.
About 100 ce. of 0.1 N sodium thiosulflLte solution.
Starch test solution.
Procedure.-l. "Introduce about 0.8 Gm. of a solid fat or about 0.3 Gm. *
of an oil, llccurately weighed, into a glass-stoppered flask or bottle of 250-ec.
capacity, dissolve it in 10 ce. of chloroform, add 25 ce. of iodohromide T.S.,
accurately measured from a burette or pipette, stopper the vessel securely,
and allow it to stand for thirty minutes t in a cool plnce protected from light."

Only 0.3 Gm. of olive oil is used in this determination, as its

iodine value is comparatively high (79 to 90).
The Pharmacopoeia directs that 0~8 Gm. be taken for solid
fats, as their iodine values are low as compared with those of the
fatty oils, the iodine value of oil of theobroma, for instance, being
33 to 38.
The oil is soluble in chloroform, and since iodine and iodine
bromide are also soluble in this solvent, it serves as a good
reaction medium.
The oil upon standing in contact with the iodobl'omide test
solution absorbs iodine. Iodine, itself, is absorbed very slowly
by the oil, but iodine is absorbed readily from bromine containing
solutions, probably through a reaction of iodine bromide, IBr,
wherein the bromine functions as a catalytic agent. The
unsaturated acids of the oleic and linoleic series present in the
olive oil, as well as their glyc:eryl esters, absorb iodine to form
addition products. Thus oleic acid C17HssCOOH takes up 2
atoms of iodine and forms the addition product cli-iodo-stearic
acid C17HssIzCOOH. The mixture, which is directed to be placed
* 0.15 to 0.18 Gm. of linseed oil, 0.18 to 0.2 Gm. of ('od liver oil, 0.8 to 1.0
Gm. of oil of theobroma.
t Allow to stand for one hour in the case of castor oil, cod liver oil or
linseed oil.
358 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

in a glass-stoppered flask (Fig. 61) to prevent the escapc of iodine

and bromine vapors, should also be protected from light to
prevent other sidc reactions from taking l)lace in which iodine is
consumed. If after standing for }~ hr. the solution is nO longer
colored brown, the amount of iodine present was insufficient to
saturate the oil.
2. "Then add in the order named 30 ce. of potassium iodide T.S. and
100 cc. of distilled water, and titrate the liberated iodine with tenth-llormal
sodillm thiosulfate, shaking thoroughly after each addition of thiosulfate.
When the iodine coior becomes quite pale, add 1 cc. of starch
T.S. and continue the titration with thiosulfate until the blue
color is discharged."

Upon the addition of the potassium iodide solu-

tion, the iodine bromide reacts with the K1,
liberating iodine until all of the bromine is used up:

K1 + IBr--+KBr + 12
An excess of K1 is added to insure the complete
FIG. 61.-
Iodine titra-
removal of the free bromine and to prevent the
tion flask.
precipitation of the iodine in the aqueous solution.
The sodium thiosulfate reacts with the excess iodine as follows:

3. "Carry out a blank test at the same time with the same quantities of
chloroform and iodobromicle solution, allowing it to stand for the same length
of time and titrating as directed. The difference in the number of cubic
cent.imeters of thiosulfate consumed by the blank test and the actual test,
multiplied by 1,269 and divided by tlle weight of sample taken gives the
Iodine Value.
"NOTE.-If the number of cubic centimeters of tenth-normal sodium
thiosulfate consumed in the actual test is less than 60 per cent of the quantity
consumed in the blank test, the determination must be repeated, using a
smaller amount of the material being assayed."

The blank test, when carried out under the same conditions
as the actual test, corrects for the presence of impmities in the
reagents, changes in volume at different temperatures, etc.,
and makes it unnecessary to know the exact nOl'l.aality of the
iodobromide test solution. The blank test should be carried out
 OONSTANTS OF FATS, FATTY OILS, WAXES, ETC. 359

in a manner ()xactly duplicating that userl in the actual test in so

far as possible, especially in the meaSllTement of the iodohromide
test solution, which has a high coefficient of expansion. It is
required that the determination be repeated if the number of
cubic centimeters of 0.1 N Na2S203 consumed in the actual test
is less than 60 per cent of the quantity consllmed in the blank
test to insure the presenee of an excess of iodine.
If in the blank test 2,1) ec. of 0.1 N Na 2S 203 are cOll:sumecl, and
in the actual test 5 cc. are required, the calculations may be
made as follows: Since 1 ee. of 0.1 N Na~S203 is equivllient to
10 ~6i~000 = 0.01269 Gm. iodine, then 25 - 5 = 20 ee. 0.1 N
Na 2S2()a equivalent to the iodine absorbed by thf) oil, and the
weight of iodine which would be absorbed by 100 Gm. of the oil
. 20 X 0.01269 20 X 1.269 . .
IS equal to 0.3 X 100 or 0.3 = 84.6, the lOdme

number of tho sample of olive oil.

TABLE LIV.-OFFICIAL SUBS'rANGES WITH THEIR IODINE NUMBERS
Iodine Number,
Substance Official Requirement
U.S.P.
Acid, oleic., ............... ,., .. ..... 85 to 95
Acid, stearic ........................ , Not morc than 4
Castor oil. ........ , .... , , .......... , . 83 to 88
Chaulmoogra oil. ................ , , , . . 93 to 104
Cod liver oiL .. , .......... , . , . . . . . . . 145 to 180
Cod liver oil, non-d('stollrinatcd ....... , 128 to 180
Corn oil. , ............ , ............. , 112 to 128
Cottonseed oil. ....... , ......... , . , , , , 105 to 114
Et.hyl chaulmoogratc .......... , . , . . . .. 90 to 100
Expressecl oil of almond .... , .. , , . . . . . . 93,to 100
Lard .................... , . . ... . . . . . . . 46 to 70
Linseed oil. ....... " ... , " ...... "'" .Not less than 170
Oil of theobroma ............... , . , , , , 35 to 40
Olive oil ........ , .......... , . , .. , . , . . 79 to 88
Prepared suet.,., , , ........ , . . . . . . . . . 33 to 48
Soap, hard (acids from) , ............ , . 83 to 92
Soap, soft (acids from) , , . , .. , ....... " Not. lesH than 170
Wool fat, . , ......... , , , ........ , . . . . 18 to 36
N.F.
Croton oil. . , .... , , .. , ... , ....... , .. . 104 t.o 110
Oil of sesame .. , .... , , ... , .. , . , ...... , 103 to 115
360 QUANTITATIVE PHAR.MACEU,!'ICAL CHEMISTRY

Questions and Problems

1. Define the term iodine vnlue.
2. Indicate by struetural formulae how iodine may add on to an unsatu-
rated fatty acid.
S. How maya determination of the iodine number be used to supplement
the information relative to the nature and purity of an oil gained through
a determination of the saponification vaille?
4.. What functions do the bromine, acetic acid, chloroform, and potassium
iodide serve iil the determination of the iodine number of an oil?
Ii. Given an unknown s[I)nple of a fatty oil. What observations, tests,
and determinations would you perform in order to establish the identity
and purity of the oil?
 CHAPTER XXIII

ASSAY OF VOLATILE OILS

Volatile oils, also known as ethereal oils or essential oils and, in

some cases, as essences, al'e generally complex products composed
of mixtures of compounds of widely variant chemical eharac~
teristics. The most important chemical components of the
official volatile oils are:
1. Hydrocarbons, occasionally of the aliphatic series, such as
heptane, myrcene, and various paraffins; but more often of the
aromatic series, e.g., pinene, camphene, limonene, bornylene,
fenchene, dipentene, sylvestrene, and phellandrene.
2. Alcohols, present both in the free state and in combination
with acids as esters. The alcohols most generally found in the
official volatile oils are li~lalool, geraniol, citronellol, terpineol,.
borneol, menthol, and santalol.
3. Aldehydes, such as benzaldehyde, cinnamic aldehyde, salicyl
aldehyde, citral, and citronellal.
4. Ketones, the most important being camphor, carvone,
fenchone, thujonc, and menthone.
5. Phenols, e.g., anethol, eugenol, carvacrol, safrol, chavicol,
and thymol.
6. Acids, sometimes present in the free state in small quanti-
ties, those occurring most commonly being acetic, propionic,
butyric, valerie, benzoic, cinllamic, and hydrocyanic acids; but
more often occurring in combination with the alcohols men-
tioned under 2 as esters, e.g., linalyl acetate, bornyl acetate, and
menthyl acetate.
7. Sulfur compounds, such as the allyl thiocyanate, found in
mustard oil.
The analysis of volatile oils for the purposes of determining
their purity and vaJue is based on the measurement of certain
physical characteristics, on the quantitative estimation of certain
components as alcohols, phenols, esters, and aldehydes, and on
361 .
362 QUANTITATIVE PHARMACEUTICAL CHEMIS,]'RY

the qualitative tests for the various substances which are com-
monly employed as adulterants. Only the official quantitative
procedures will be discussed here.
Methods of General Application.-The purity and quality of
volatile oils may be judged to Rome extent by their appearance,
odor, color, etc.; but the information gained from the determina-
tion of the specific gravity, rotatory power, refract.ive index,
solidifying point, solubility, and behavior on distillation is of
much greater importance.
Specific Gravity.-The specific gravity of a volatile oil may be
determined with the Westphal balance or pycnometer, t.he
latter being the more accurate method of the two, and expressed
as the ratio of the weight of the volume of oil t.o that of an equal
volume of pure water when both are determined at 250. (see
page 217).
The specific gravities of the official volatile oils vary approxi-
mately between 0.84 and 1.2. Those oils which are lighter
than water are usually rich in hydrocarbons, alcohols, esters,
aldehydes, and ketones, such as oil of orange, caraway, coriander,
lemon, turpentine, and rosemary. Oils the specific gravities of
which approach or exceed 1.0 usually contain chiefly phenols,
phenolic derivatives, or certain esters, e.g., oil of anise, cinnamon,
clove, sassafras, and mustard.
The specific gravity of any volatile oil is not absolutely con-
stant, sincc it is influenced by such actors as the maturity of the
plant from which the oil is obtained, as well as the method of
preparation, purification, and age of the oil.
Rotatory Power.-The rotatory power of a volatile oil is
generally measured with a Laurent half-shadow polarimeter,
according to the procedure described on page 247, using sodium
light and a tube 10 cm. long, but for highly colored oils, tubes 5
or even 2.5 cm. long may be used. The observation of the
optical activities of the official essential oils should be made at
25C. Slight deviations from this temperature do not greatly
affect the rotatory power of a volatile oil, except in the case of oil
of lemon and oil of orange.
The rotatory power of some of the volatile oils varies within
relatively wide limits. This determination should never be
omitted in their examination, however, since it frequently serves
 ASSAY OF VOLATILE OILS 363

as a valuable means of detecting adulteration with inactive sub-

stances, such as alcohol, or with substances pf different rotatory
power from that of the oil being examined, e.g., oil of lemon
(+57 to +64) adulterated with oil of turpentine (about +25 to
-40).
Refractive Index.-The refractive index of a volatile oil is
most commonly determined by means of an Abbe refractometer,
as described on page 240. The measurement of the refractive
indices of all of the official volatile oils should be performed at
20 o e. The index of refraction does not vary greatly with
different official volatile oils, the values being between about
1.46 and 1.61 at 20 0 e. In some cases, this determination may,
however, serve for the detection of extraneous matter.
Congealing Point.-The congealing point of a volatile oil is
determined by the method described on page 231. Most essen-
tial oils solidify only at low temperatures, eonsequently in
practice, this detcrmination is earried out with only a few oils,
such as oil of anise and oil of fennel, which contain large amounts
of the readily crystallizable constituent anethol. The higher
the congealing point of these oils the more they are valued.
An abnormally low congealing point of an essential oil indicates
the partial removal of the characteristic constituent for which
the oil is valued or the addition of extraneous matter, such as
alcohol.
Distilling Point.-The distilling point of a volatile oil is
determined by the general method described on page 234.
Volatile oils which are composed of mixtures of hydrocarbons,
alcohols, esters, etc., do not have a fixed boiling point but boil
between certain limits of temperature, frequently separated
widely. Consequently, the official standards usually designate
the tempcl'l1ture or range of temperature at which a definite
percentage of the oil distils; e.g., 90 per cent of oil of turpentine
should distil between 154 and 170 o e., and less than 10 per cent of
oil of dwarf pine needles should distil below 165e.
Fractional Distillation.-Fractional distillation is resorted to
occasionally in the official tests for purity of volatile oils. It
serves either to separate the various components of volatile oils
or to detect adulteration. Thus, alcohol and petroleum ether,
both of which boil below lOOoe., may be separated and identified
364 qUANTITATIVE PHARMACEUTICAL CIIEllfISTRY

in this way. The addition of oil of turpentine and other adulter-

ants may be detected by collecting various fractions of the oil
and determining the rotary power and refractive index of
each fraction; e.g., see Oil of Orange, U.S.P., pH,ge 250.
Solubility.-The volatile oils are generally soluble in organic
solvents, such as absolute alcohol, ether, chloroform, benzene,
carbon disulfide, etc. They dissolve more or less readily in
dilute alcohol according to the nature of their components. Oils
containing a large percentage of oxygenated substances often
produce turbid solutions with petroleum ether 01' carbon disul-
fide due to the separation of water, sIllall quantities of which
are dissolved in such oils.
Practically all of the volatile oils exhibit almost constant
solubility in 90, 80, or 70 pel' cent alcohol. Consequently, the
test of the solubility of a volatile oil in dilute alcohol frequently
gives valuable data relative to its purity, since the commonly
used adulterants, oil of turpentine, petroleum oils, and fatty
oils, are but slightly soluble in 80 or 70 pel' cent alcohol.
Assay for Ester Content.-A number of the official volatile
oils are e~aluated on the basis of their ester content. The esters
found are 1l10stlythe acctates of alcohols of the formula C 1oH 170H,
e.g., bornool, geraniol, linalool, and terpineol; C1oH100H, such
as menthol 01' citronelloli or C lIiH 23 0H, e.g., santalol. When
these esters are refluxed with alcoholic potassium hydroxide,
they are saponified and yield the free alcohol and the potassium
salt corresponding to the acid component of the ester.
The number of milligrams of KOR required to saponify 1 Gm.
of the oil represents the ester value 01' ester number of the oil.
The ester number for any given oil may be subject to considerable
variation, but the minimum requirement set by the official
standards must be complied with when the oil is represented as a
product conforming to the U.S.P. 01' N.F. standards. The
determination of the total esters when taken in conjunction with
the official tests for purity serves to detect adulteration
and to establish the quality of those oils valued for their ester
content.
The ester values of oils which contain appreciable amounts of
aldehydes or phenols cannot be estimated accurately by saponifi-
cation with an alkali, since variable amounts of the latter are
 ASSAY OF VOLA1'ILE OILS 365

consumed by reaction with the aldehydes and phenols, or their

decomposition products.

Exercise 104

Object.-To Prepare 0.5 N Alcoholic Potassium Hydroxide.

Materials Required.-2.5 Gm. of lelld acetate.
1,500 ce. of alcohol.
40 Gm. of potassium hydroxide.
Procedure.-l. "Dissolve 2.5 GIll. of lead acetate in 5 ce. of distilled
water, add the solution to 1000 cc. of alcohol contained in a glass-stoppered
bottle and mix thoroughly. Dissolve 5 Gm. of potassium hydroxide in
25 cc. of warm alcohol, cool the solution, and add it slowly, without stirring,
to the alcoholic solution of lead acetate. After one hour shake the mixture
vigorously, allow it to stand over night, decant the clear liquid ::md recover
the alcohol by distillation."

The potassium hydroxide converts any acids present into non-

volatile salts and causes the polymerization of aldehydes. The
lead acetate is decomposed by the ROn with the formation of
lead oxide and potassium acetate. The lead forms insoluble and
non-volatile compounds with any sulfur compounds present.
The lead oxide carries down the polymerized aldehydes when it
is precipitated so that it is easy to decant the clear supernatant
liquid.
2. "Dissolve about 35 Gm. of pota:~sium hydroxide in 20 cc. of distilled
water and add sufficient. aldehyde-free alcohol to make 1000 cc. Allow the
solution to stand in a tightly-stoppered bottle (using either glass or rubber
stopper) for twenty-four houl's. Then quickly docant the clear super-
natant liquid into a bottle provided with a well-fitting rubber-stopper and
standardize as follows:"

Traces of aldehydes remaining in the alcoholic potassium

hydroxide solution are polymerized and settle out when the
mixture is allowed to stand. The rubber stopper used should
be washed with a small quantity of the alkaline alcoholic solution
before it is inserted into the mouth of the bottle to dissolve
sulfur and any other material which might affect the concen-
tration of the solution.
3. "Measure accurately, from a burette, about 25 cc. of half-normal
hydrochloric acid. Dilute with 50 ceo of distilled water, add 2 drops of
phenolphthalein T.S., and titrate with the alcoholic potassium hydroxide
366 QUANTITA'l.'IVE PHARMACEUTICAL CHEMISTRY

~olut,jon until a permanent, pale pink ~olor is produced. Calculate the

normality."
The titratiop. is carried out as described on page 81. The
prepared solution should be preserved in tightly stoppered bottles
to prevent changes in the concentration of alkali through absorp-
tion of CO 2 Well-fitting rubber stoppers lllay be used. It
should also be protected from light to prevent the development
of reddish colored compounds which interfere with the correct
determination of the end point during titration. The solution
should be restandardized whenever it is used, and it should
alwa,ys be measured at the same temperature at which it was
standardized, since alcohol has a high coefficient of expansion.
Exercise 105

Object.-Assay of Oil of Peppermint for Total Esters.

Materials Required.-lO ce. of oil of peppermint.
25 ce. of 0.5 N alcoholic KOH.
About 25 ce. of 0.5 N H 2S0 4
Procedure.-l. "Place 10 ce. of oil of peppermint in a tared, 125 cc.
Erlenmeyer flask, and weigh accurately. Add 25 ce. of half-normal alcoholic
potassium hydroxide, connect the flask with a rcflux condenser, and boil
the mixture on a water bath for OIle hour."

Instead of weighing the oil directly in a previously tared

Erlenmeyer flask, it may be weighed in a weighing bottle, and the
weighing bottle with its contents may then be transferred into the
Erlenmeyer flask. The esters of oil of peppermint, which con-
sist chiefly of menthyl acetate, are sa,ponified by the alkali when
the mixture is refiuxed, forming free menthol and potassium
acetate. Alcoholic potassium hydroxide is used because the
oil is soluble in strong alcoholic solution.

2. "Allow the mixture to cool, disconnect the flask from the condenser,
and titrate the excess of alkali with half-normal sulfuric acid, using 10
 ASSAY OF VOLATILE OILS 367
drops of phenolphthalein T.S. as the indicator. Subtract the number of
cc. of half-normal sulfuric acid required for neutralization from the 25 ce.
of half-normal alcoholic potassium hydroxide taken, multiply the difference
by 9.912, and divide this product by the weight of oil of peppermint taken,
the result shows the per cent of esters calculated as menthyl acetate."

The 0.5 N acid neutralizes the excess 0.5 N alkali:

The difference between the number of cubic centimeters of 0.5 N

KOH added and the number of cubic centimeters of 0.5 N H 2SO J
required in the titration represents the alkali consumed in the
saponification of the esters present in the oil; e.g., if a 9 Gm.
sample of the oil refluxed with 25 cc. of 0.5 N KOH required 20.5
cc. of 0.5 N H 2S0 4 to neutralize the excess alkali, the amount of
standard alkali Rolution consumed in the saponification of the
esters present in the oil would be 25 - 20.5 = 4.5 cc. The per-
centage of total esters present in the oil calculated as menthyl
acetate is found as follows: 4.5 X 9.912 = 4.96 per cent menthyl
9
acetate. The factor 9.912 is derived from the molecular weight of
menthyl acetate, 198.24. Thus, 1,000 cc. of N KOH is equivalent
to 198.24 Gm. of menthyl acetate, and 1 cc. of 0.5 N KOH is
equivalent to 2 ~8i~0400 = 0.09912 Gm. menthyl acetate. This
equivalent is multiplied by 100 so that. the factor 9.912 when
multiplied by the number of cubic centimeters of 0.5 N alkali con-
sumed in the assay and divided by the weight of the sample of
oil used will give the percentage of esters directly.

Questions and Problems

1. What effect would the presence of free acids in the oil have upon the
result. of the determination of total esters?
2. Why is an alcoholic rather than an aqueous solution of KOH used to
saponify the esters?
3. What factor would be necessary for the ester in order to obtain the
per cent of total esters directly as described above if the chief ester present
in an oil was bornyl acetate; linalyl acetate? Show how these factors are
derived.
4. Note the equivalents given in the following table for linalyl acetate in
oil of lavender, U.S.P., and in oil of bergamot, N.F. Which is correct?
368 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

TABLB LV.-OFFICIAL VOLATILE OILS ASSAYED FOR THEIR ESTER CONTENT

Amount Equivalent of
Official requirement,
Oil used, 1 CC. of 0.5 N
per cent
Gm . KOH, Gm.
.-
U.S.P.
Methyl salicylate ... _.. 2 0.07603 Methyl salicylate
C OH 4 (OH)C0 2CH a = 98
Oil of dwarf pine
needles ............. 10 0.09811 Bornyl acetate
CloHl7C2Ha02 = 5
Oil of la vender ........ 5 0.09811 Linalyl acetate
ClOH17C2Ha02 = 30
Oil of peppermint ..... 10 0.09912 Menthyl acetate
C IOH 10C.H aO. = 5
Oil of rosemary ........ 10 0.09811 Bornyl acetate
CloH17C2Ha02 = 2.5
N.F.
Oil of bergamot ....... 2 0.09808 Linalyl acetate
C 1o H 17C,H aO. = 36

Assay for Alcohol Content.-The alcohols present in volatile

oils, such as mcnthol and borneol, occur both free and
combined as esters. Usually, the determination of

\
I the total alcohol, occurring free and combined, is
sufficicnt to establish the purity and value of an oil
with respect to its content of alcoholic constituents.
The total alcohols present in any given oil are
determined by transforming the free alcohols into the
corresponding acetates by boiling the oil with acetic
anhydride in an acetylization flask (Fig. 62) and then
determining the saponification value of the acetylized
product. When the alcohol occ(Jrs partly free and
partly combined in the form of an ester, a correction
factor must be employed. The method of determin-
ing the total alcohol content of volatile oils is
FIG. 62.-
illustrated in the following exercise:
Acetyli-
zation flask Exercise 106
wi th air
condenser. Object.-Assay of Oil of Peppermint for Total
Menthol.
Materials Requited.-l0 ce. of oil of peppermint.
10 cc. of acetic anhydride.
 ASS,AY OF VOLA'l'ILE OILS 369
10m. of anhYdrous sodium acetate.
3 Om. of monohydrated sodium carbonate.
About 0.2 Om. of fused calcium chloride.
50 cc. of 0.5 N alcoholic KOH.
50 ce. of 0.5 N H 2S04.
An acetylization flask of about 100 cc. capacity with till air condenser
about 100 em. long.
Procedure.-l. "Place 10 ce. of Oil of Peppermint in an aeetylization
flask of 100 ce. capacity, add 10 ce. of acctic anhydride amI 1 Gm. of
powdered anhydrous sodium acetate. Boil the mixture gently for olle hour,
cool, disconnect the flask from the condenser, transfer the mixture to [1,
small separator, rinsing the acetylizatiol1 flask with three successive 5 ce.
portions of wann distilled water, and add the rinsings to the scparator."

The acetylization flask should be supported on an asbestos

board while heating to prevent decomposition of the oil. The
acetic anhydride reacts with the menthol, forming menthyl
acetate and acetic acid:

-OR + 0",
'\. O-OHa
/
.-}
9' 0
'\.
-O-C-CHa + CHaCOOH
C-CHa
I'
o
Anhydrous sodium acetate is added to absorb any traces of
water liberated in the side reaction:

6~
-OR
0
'\.
+ RQ-C-CHa-).
~
o
-O-C-eHa + HGH

2. "When the liquids have completely sepltmted, reject the aqueous layer,
and wash the reIllaining oil with successive portions of sodium carbonate
T.S., diluted with an equal volume of distilled water, until the washing is
alkaline to 2 drops of phenolphthalein T.S. Dry the r~s\llting Oil with
anhydrous sodium sulfate (prepared by drying sodium sulfate to COD stant
weight at noc. and powdering), and filter it."
370 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The oil is washed with sodium carbonate test solution (12.5 Gm.
of Na 2CO a.H 20 in sufficient distilled water to make 100 cc.) to
neutralize the excess acetic acid. The washed oil is then dried
over anhydrous sodium sulfate to remove the small amounts of
alkaline sodium carbonate solution which it contains.
3. "Transfer 5 ec. of the dry acetylized oil to a tared, 100-ce. Erlenmeyer
flask, llote its exact weight, add 50 ce. of half-normal alcoholic potassium
hydroxide, connect the flask with a reflux condenser, and boil the mixture
on a water bath for one hour."

The 0.5 N ROH saponifies the menthyl acetate, forming men-

thol and potassium acetate in the same manner as in the assay for
total esters. The liberated menthol dissolves in the alcoholic
solution.
4. "Allow the mixture to cool, disconnect the flask from the condenser,
and titrate the excess of alkali with half-normal Bulimic acid, using 10 drops
of phenolphthalein T.S. as the indiw1tor. Calculate the per cent of menthol
by the following form1lla:

Per cent of total menthol in the Oil tested = B _::\~ ~8~~21)

X [1 - (E X 0.0021)J.

A is the result obtained by subtracting the number of ee. of half-normal

sulfuric acid required in the above titration from the number of cc. of half~
normal alcoholic potassium hydroxide originally taken, B is the weight of
acetylized oil taken, and E is the percentage of esters caleulo,ted as menthyl
acetate (CIUH19.C2Ha02)."

The number of cubic centimeters of 0.5 N KOH consumed in

the 'saponification of a definite weight of the acetylized oil is
equal to the difference between the number of cubic centimeters
of 0.5 N l{OH added and the number of cubic centimeters of 0.5 N
H 2S0 4 required in the residual titration. The formula used in
the calculation is derived as follows: The molecular weight of
menthol is 156.16. One cubic centimeter of 0.5 N ROH is
equivalent to 2 ~6i~0600 = 0.07808 Gm. menthol. In order that
the result may be read directly in terms of per cellt, the factor
0.07808 is multiplied by 100, giving 7.808. B - (A X 0.021)
corrects for the weight of acetyl radical in the sample of acetyl-
ized oil, the factor 0.021 being the amount of acetyl radical
equivalent to 1 cc. of 0.5 N KOH. The latter is obtained from
 ASSAY OF VOLATILE OILS~ 371

198.17' (molecular weight of menthyl acetate) - 156.16 (molec-

ular weight of menthol) = 42.01 (molecular weight of the acetyl
radical). Therefore, 1,000 cc. of N KOH is equivalent to 42.03
Gm. of. acetyl radical, and 1 cc. of 0.5 N KOH is equivalent to
42 '
2 X 1,000 = 0.021 Gm. acetyl radical. (A X 0.021), therefore,
represents the weight of acetyl radical to be deducted from B, the
weight.of acetylized oil taken.
Since the original oil contained some menthol combined as ester
and calculated as menthyl acetate, it would be incorrect to sub-
tract all of the weight of the acetyl radical from B. A correction
for the weight of acetyl radical in the original oil is consequently
applied by multiplying the per cent found from the fonnula
B ! (~ ~8g.~21) by the correction formula [1 - (E X 0.0021)].
The factor 0.0021 is the ratio of the weight of acetyl radical to
that of menthyl acetate 42/198.17 = 0.21. If E, the per cent of
total esters determined by the preceding assay, is found to be 5,
then the formula would be [1 - (0.05 X 0.21)1 or [1 - (5 X
0.0021)1: Applying this correction, the formula becomes
A X 7.808
B _ (A X 0.021) X [1 - (5 X 0.0021)] = per cent total menthol.
The importance of the correction [1 - (0.05 X 0.21)] may be
seen from the following example: A sample of oil of peppermint
found to contain 5 per cent of total esters calculated as menthyl
acetate and 50 per cent of total menthol as calculated from the
A X 7.808
formula, per cent total menthol = B _ (A. X 0.021)' gave the
following percentage of total menthol when the correction was
applied: 50 X [1 - (5 X 0.0021)] = 50 X (1 - 0.0105) = 50 X
0.9895 = 49.475 per cent menthol free and as esters. The
correction involves about one-half of 1 per cent menthol or 50 -
49.475 = 0.525 per cent.
A similar correction is applied in the assay of oil of rosemary,
but none is given in the assay of oil of santa!, since the santalol
does not occur combined in the form of esters.
Questions and Problems
1. Having determined the ester value and the percentage of total borneol
present in oil of rosemary, how could the percentage of free borneol be
found? Give an example.
372 QUAN'l'ITtl.TIVE PHARM1"1CEU'l'ICAL CHEMISTRY

2. Write the re~ctions which take place in the assay of oil of rosemary for
total borneol.
3. If oil of peppermint containing 7.2 per cent of total esters is found to
eontain 51.5 per cent of total menthol (uncorrected), what change in the
result would the correction 1 - (E X 0.0021) cause? Is this correction
import~nt? .
TABLE LVI.-OFFICIAL VOLATILE OILS ASSAYED ~'Oll THEIR ALCOHOL
CONTEl<1T

Amoun. of Equivnlent
IIcc.yJized of 1 CC. of Official roquire-
Oil Correction factor
oil used, 0.5 N IWH, lnent, per cent
Gm, Gm.
- -~~--~<--

U.S.P.
Oil of pepp"rmint. .. 5 0.07808 1 - (E X 0,0021) Menthol
OlOlh,OH = 50
Oil of ro,emury ..... 5 0.0771 1 - (E X 0.0021) . Borneol
ClOHIlOH = 10
Oil of sun!nl. ....... 5 O.W)1 None BuntaInl
C"n"OH = 90

Assay for Aldehyde Content.-Aldehydes, like ketones, form

addition products with certain reagents, and this property is
utilized in their assay. When no other constituents are pres-
ent which react with the reagent, the assay for aldehydes may be
performed by the sulfite method described under the assay for
ketones (page 375). Thus the cinnamic aldehyde content of the
offieial oil of cinnamon is determined by this method, the basic
reaction being
o
./ NaO 0
~s~ ~
O
CR CH-C "'H +2
\,. Na 0

+ 2NaOH
 ASSAY OF YOLAl'ILE OILS 373

The sulfite addition product dissolves in water leaNing the 11on-

aldehyde constituents as a water-insoluble layer. The volume
of this wll-ter-insoluble layer is then measured in a cassia fhtsk,
(Fig. 63).
The cassia flask is a glass flask of about 100 cc. captlcity with a
long, narrow neck graduated to 10 CC. in 0.1 cc. divisions. In the
assay for aldehydes and ketones by the sulfite method or bisul-
fite method, the portion of the oil which does not react to form
a water-soluble addition product rIses to the surface, and when
the flask is filled, this oily layer rises into the graduated neck
of the cassia flask where its volume can be meas-
ured. The volume of the residual oily layer
subtracted from the volume of the sample of oil used
represents the volume of the aldehyde constituents
which formed a water-soluble addition product with
the sulfite.
Those volatile oils which contain aldehydes in
very small arnounts, such as oil of lemon, or which
contain other constituents that form water-soluble
addition products with sodium sulfite and bisulfite
cannot be assayed accurately by this method. In FIG.53.-
Cassia
these cases, the aldehydes are generally determined flask.
by the phenylhydrazine method (sec assay of
benzaldehyde, N.F.) or by the hydroxylamine method described
in the following exercise.
Exercise 107

Object.-Assayof Oil of Bitter Almond for Benzaldehyde.

Materials Required.-l cc. of oil of hitter almond.
3.5 Gm. of hydl'oxylmnille hydrochloride.
100 cc. of 60 per cent alcohol.
Bromphenol blue '1:'.S.
0.5 N alcoholic KOH.
Procedure.-l. "Ad(l 0.1 cc. of bromphenol hlue T.S. to 25 cc. of a solu-
tion made by dissolving 3.5 Gm. of hydroxylamine hydrochloride in sufficient
60 pel' cent alcohol to make 100 ce., and titrate with half-normal alcoholie
potassium hydroxide to the production of a greenish-blue color."

Hydroxylamine hydrochloride reacts as a neutral salt when

bromphenol blue is used as the indicator. Oonsequently, upon
374 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

titration of the solution of hydroxylamine hydrochloride with

0.5 N KOH, the free Hel present in the solution is neutralized.
If an indicator which changes color on the alkaline side of the
pH scale, such as phenolphthalein, is used, the end point will be
obtained when the free and combined acid has been neutralized
by the alkali.
2. "Pour this mixture into a flask, fitted with a glass stopper, and con-
taining about 1 Gm. of Oil of Bitter Almond, accurately weighed.. Shake
well and then titrate with half-normal alcoholic potassium hydroxide until
t.he yellow color changes to greenish-blne. Continue shaking and titrating
until the greenish-blue color is permanent. Each cubic centimcter of half-
normal alcoholic potassium hydroxide is equivalent to 0.05303 Gill. of
C 6H 6 .CHO."

Aldoximes are formed when a cold aqueous or hydro alcoholic

solution of hydroxylamine hydrochloride is added to an aldehyde
in the presence of an equivalent amount of KOH. The equations
for the reactions that take place may be represented as follows:

The basic hydroxylamine reacts with the benzaldehyde to form an

unstable addition product which loses water and readily forms the
oxime.

It is apparent from the equations given that the assay is based

upon the determination of the amount of HCl obtained from the
quantity of hydroxylamine hydrochloride equivalent to the
benzaldehyde contained in the sample.
The HeN present in the oil of bitter almond does not influence
the accuracy of the assay. Hydrocyanic acid is a very weak acid.
In the presence of a strong acid, such as HOI, the common
hydrogen ion represses the dissociation of the weak acid, HCN,
so that at the end point, which in this case is at a pH of about
4.2, none of the HCN enters into the reaction.
 ASSA Y OF VOLA. TILE OILS 375
TABLE L VII.-OFE'ICIAL VOLATILE OILS ASSAYED FOR ALDEHYDE
CONTENT

Amount Official requirement,

Oil Method
used per cent

U.S.P.
Oil of bitter al-
mond ......... 1 Gm. Hydroxylmnine Benzaldehyde = 95
Oil of cillnllmol~ .. 10 cc. Sulfite Cillnmnic 1l14ehydo = 80
N.F.
Benzllldehyde .. , . 1 Gm. Phenylhydrazine Benzaldehyde = 85

Questions and Problems

1. Show how the equivalent factor for benzaldehyde is derived.
2. Why is the titration carried out in a hydroaleoholie solution?
3. Look up the method of assay for benzaldehyde in the National
Formulary and write equations for the reactions that take place,
4. Does the presenee of HCN eause an appreciable error in the assay of
oil of bitter almond for benzaldehyde content?

Assay for Ketone Content.-Only two of the official volatile

oils are assayed for their ketone content. In each case, the
determination is made by the so-called sulfite method. This
method is based on the property of ketones to combine with
sodium sulfite to form water-soluble addition compounds.
Exercise 108
Object.-Assay of Oil of Caraway.
Materials Required.-A 100 ce. cassia flask.
10 cc. of oil of caraway.
100 cc. of saturated sodium sulfite solution (40 Gm. in 100 ce.).
100 cc. of sodium bisulfite solution (5 per cent).
Procedure.-l. "Introduce exaetly 10 ce. of Oil of Caraway, accurately
me!Lsul'ed, into a cassia flask, and add 50 cc. of a saturated solution of sodium
sulfite which has been carefully rendered neutral to 2 drops of phenol-
phthalein T.S. by means of a saturated sodium bisulfite solution. Heat the
flask in boiling water and shake it repeatedly, neutralizing the mixture from
time to time by the addition of a few drops of the saturated sodium bisulfite
solution. "
Carvone, the principal ketone present in the oil, reacts with the
sodium sulfite, forming a water-soluble addition product as
follows:
370 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

This addition product is hydrolyzed in part by the water with the

formation of sodium hydroxide, viz.:

The presence of free alkali in the aqueous layer renders other

components of the oil, such as phenols, water-soluble and also
tends to reverse the addition reaction. To prevent this the
sodium hydroxide is neutralized by the addition of sodium acid
sulfite:

2. "When no coloration appear;; upon adding a few more drops of phenol-

phthalein T.S. and heating for 15 minutes, cool the mixture to room tem-
perature, and, when the liquids have separated eompletely, add sufficient
sodium sulfite solution to raise the lower limit of the oily layer within the
graduated portion of the neck of the flask. Note the volume of the residual
oily liquid. This volume does not exceed 5 cc., indicating the presence in
the Oil of not less than 50 per cent, by volume, of carvone."

The difference between the volume of oil used as the sample and
the volume of the oily layer which remains insoluble represents
the carvone which dissolved in the aqueous layer. If drops of
oil adhere to the walls of the flask, they may be made to rise into
the neck by gently tapping and rotating it. If the residual
liquid measures 4.5 cc., the percentage of carvone by volume in
the oil would be 10 ~o 4.5 X 100 = 55 per cent.
The only other official oil evaluated for its ketone content is oil
of spearmint. It is assayed in exactly the same way as is oil of
 ASSAY OF VOl.J.11'ILE OILS 377

caraway, and it is required to contain not less than 50 per cent by

volume of carvone.
Questions and Problems
1. Ketones may also be estimated in volatile oils by the use of sodium
bisulfite alone. Write the reaction between carvone and sodium bisulfite.
2. What substances, if present, in the oil would cause an error in the
results of the sulfite method of assay for ketones?
3. How could the carvone be isolated from the aqueous solution?
Assay for Phenol Content.-Volatile oils whieh contain phenols
when shaken with solutions of sodium or potassium hydroxide
diminish in volume due to the ready solubility of the phenol
constituents in alkali; the non-phenolic portion of the oil remains
undissolved.
Use is made of this property of phenols in the official assay
methods. In these determinations a flask with a graduated neck
like that employed in the estimation of aldehydes (see page 373)
is used. In this flask, an accurately measured volume of the oil is
shaken vigorously with a portion of the caustic alkali, and a
further quantity of alkali is then added to bring the undissolved
oil into the graduated Heck of the flask. When the liquids have
separated and become clear, the volume of the oil insoluble in
alkali is read off, the volume of the phenol being determined by
difference.
Exercise 109
Object.-Assay of Oil of Clove.
Materials Required.:........l0 ce. of oil of clove.
100 ee. of I{OH solution (6.5 Gm. in 100 ce.).
A eltssia flask.
Procedure.-l. "Place 10 ce.' of Oil of Clove, measured from It pipette,
in a 100-ce. cassia flask, add 50 ce. of normal potassium hydroxide solution,
shake the mixture for five minutes, and heat. It for ten minutes in a bath
containing boiling water. Remove it from the bath, and eool to room
temperature."
TheKOH reacts with the phenol, eugenol, as follows:
-OH -OK
OR
O-00R, a-O ,
'~OR'-OR OR, + KOR~ -OR,-OR CR, + ROR
378 QUANTITATIVE PHARMACEU7'ICAL CHEMISTRY

The mixture is heated on a water bath for 10 min. to saponify

any aceteugenol present.
2. "When the liquids have separated completely, lldd sufficient. normal
potassium hydroxide to raise the lower limit of the oily layer within the
graduated portion of the neck: the volume of the oily layer does not exceed
1.8 ce., indicating the presence in the Oil of not less than 82 per cent by
volume of engenol (C,oH120~).JJ

The potassium eugenol compound dissolves in the water leaving

undissolved hydrocarbons, ketones, etc., which rise into the
graduated neck of the flask and are measured. If stronger solu-
tions are employed, the results obtained are too high, since the
strong alkali exercises some solvent action on the non-phenolic
constituents, particularly on oxygenated compounds. The vol-
ume of the undissolved liquid subtracted from the volume of oil
used as the sample gives the volume of eugenol which dissolved in
the alkali; e.g., if a 10 cc. sample of oil yields 1.6 cc. of residual
liquid, then 10 - 1.6 = 8.4 cc. eugenol dissolved, and the per-
centage of eugenol III th I e'IS 8.4
e sarnp TO x 100 = 84 per cent.
Questions and Problems
1. Is the assay of phenols, as given above, an exact method?
2. How could the eugenol be liberated from the alkaline solution?
3. If a part of the eugenol is present combined as esters, is that part
estimated in the determination?

TABLE LVIII.-OFFICIAL VOLA'rJLE OILS ASSAYED FOR PHENOL CONTENT

Amount Alkali Official requirement,

Oil
used, cc. used per cent

U.S.P.
Oil of clove ............... 10 KOH Eugenol C IDH 120. = 82
N.F.
Oil of myrcia ............ , . 10 KOH Phenols = 50 to 60
Oil of pimenta ............ 10 KOH Eugenol CloR,.O. = 65
Oil of thyme .............. 10 KOH Phenols = 20

Assay for Hydrocyanic Acid Content.-Only one official vola-

tile oil is assayed for its hydrocyanic acid content, namely, oil of
bitter almond.
 ASSAY OF VOLATILE OILS 379

Exercise 110
Object.-Assay of Oil of Bitter Almond for Hydrocyanic Acid.
Materials Required.-0.75 Gm. of magnesium sulfate.
0.5 N sodium hydroxide.
PotassiuIU chromate indicator (10 Gm. in 100 ce. of watcr).
0.1 N silver nitrate.
Procedure.-l. "Dissolve 0.75 Gm. of magnesium sulfate in 45 cc. of
distilled water, add 5 ce. of half-normal socijum hydroxide and 2 drops of
potassium chromate T.S., and titrate the solution with tenth-nornud silver
nitrate to the production of l1. permanent reddish tint."
The magnesium sulfate reacts with the sodium hydrm,,-i.de,
forming magnesium hydroxide and sodium sulfate:
MgS0 4 + 2NaOH~Mg(OH)2 + Na S04
2

The resulting mixture is titrated with 0.1 N silver nitrate to the

appearance of a permanent end point to correct for the presence
of impurities in the reagents used which would react with the
silver nitrate in the next step of the assay.
2. "Pour this mixture into a 100 ee. flask containing about 1 Gm. of
Oil of Bitter Almond, accurately weighed, mix well, and titrate again with
tenth-normal eilver nitrate until a fell tint, which does not disappear 011
shaking, is reproduced. Conduct this titration as rapidly as possible.
Each cubic centimeter of tenth-normal silver nitrate corres~onds to 0.002702
GIll. of HCN."
The freshly precipitated Mg(OH)2 splits up the cyanhydrin
present in the oil and at the same time furnishes a white back-
ground against which the end point of titration may be observed.
The reaction may be represented as follows:
H
/

20 -C-OH

"C_N + Mg(OH),~
H
/

2
o C'\-
0 + Mg(CNh + 2HOH
380 QUAN'l'I'l'ATIVE PHARM.ACEUTICAL CJlEJ}!lSTRY

Upon titration with 0.1 N silver nitrate, silver cyanide is formed:

Mg(CN)2 + 2AgNOr---72AgCN + Mg(NOah
When all of the alkali cyanide present. in the mixture is consumed
in the reaction with the silver nitrate, the end point is shown by
the formation of the red-colored precipitate of silver chromate:

The flask should be shaken continuously during the titration to

bring the oil into intimate contact with the mixture. The titra-
tion is directed to be conducted as rapidly as possible, because
HeN decomposes in the presence of water and alkalies with the
formation of ammonia, formic acid, etc.
Each cubic centimeter of 0.1 N AgNO a consumed in the final
titration is equivalent to 10 ~~~ooo = 0.002702 Gm. HCN where
27.02 is the molecular weight of hyclroeyrmic acid.

Questions and Problems

1. Why is the magnesium sulfate added?
2. From the quantities of MgSO, and NaOH used, calculate whether
any NaOH as such will be present in the reaction mixture.
3. Indicate by reactions how HeN may be hydrolyzed in the presence
of alkalies with the formation of formatcs and ammonia.

Assay of Oil of Chenopodium for Ascaridol Content.-The

principal constituent of oil of chenopodium from a pharmaceutical
standpoint is ascaridol, an organie peroxide, which is valued for its
anthelminthic properties. Various chemical and biological
methods of evaluating the oil have been proposed, but the official
method, which is based on the solubility of ascaridol in acetic
acid, is probably as good as any, although it should be regarded as
approximate only.

Exercise 111
Object.-Assayof Oil of Chenopodium.
Materials Required.-lO cc. of oil of chenopodium.
60 cc. of glacial acetic acid.
A cassia flask.
 ASSAY OP VOLA'I'lLE OILS 381
Procedure.-l. "Pl!Lce 10 ce. of Oil of Chenoput\iulll, measured from a
pipette, in a 100 ce. cassia flask, add 50 ce. of a solutiull of rwctic l1eid, rU!lcic
by diluting 60 cc. of glacial acetic acid with distilled water, to measure
100 cc."

The ascaridol in the oil diHRolves in the 60 per cent acetie acid.
The exact nature of the reaction that occurs is not known, but it
may be represented as fo11o,,,s:

Any variation in the strength of the acetic acid employed causes

variable amounts of the constituents of the oil (decomposition and
reduction products of ascaridol) to dissolve in the acetic acid;
consequently, the acetic acid solution used must be of exact.ly
60 pel' cent strength.
2. "Shake the mixture continnously and vigorously for a period of five
minutes and add sufficient of the acetic acid solution to mi;;e the lowcr limit
of the oily layer within the graduated portion of t,he neck. When the
liquids have completely sep!lratcd so that a sharp meniscus is discernihle,
note the volume of the oily byer, at 25C. It measures not more than 4 ce.
and not less than 2 cc., indicating not less than 60 per cent and not more than
'I> 80 per cent, by volume, of the acetic-acid soluble fraction of the Oil."

The constituents soluble in the acetic acid solution dissolve,

leaving an oily layer composed chiefly of p-cymene and other
terpene hydrocarbons. Acetic aci~l solution is used to raise
the lower limit of the oily layer within the graduated portion
of the neck of the flask, because dilution with water would change
the concentration of the acetic acid and cause some of the
ascaridol to separate out with the oily liquid.
The difference between the volume of the sample useel and the
volume of the residual oily layer is considered to be asearidoL
Thus, a 10 ce. portion of the oil, 3 cc. of which remained undis-
solved, would contain 'J1 0 X 100 = 70 per cent ascarielol.
 382 QUANTITATIVE PHA_RMACEUTICAL CHEMISTRY

Questions and Pr()blems

1. I.ook up the reaction of organic peroxides with ucids in a textbook on

organic chemistry and note whet.her the probable reaction indicated con-
fOI'Ills to the information obtained.
2. Why should this lIssay be considered as only a close approximation?

Assay for Allyl Isothiocyanate.-Volatile oil of mustard is the

only official oil assayed for its content of allyl isothiocyanate.
Black mustard, which is also official, is assayed in a similar
manner, the volatile oil bcing separn,ted by distillation and its
allyl isothiocyanate content determined.

Exercise 112

Object.-Assay of Oil of Mustard.

Materials Required.-4 cc. of volatile oil of mustard.
100 cc. of alcohol.
50 ce. of 0.1 N AgNO s solution.
5 cc. of 10 per cent ammonia solution.
5 cc. of nitric acid.
2 cc. of ferric ammonium indicator solution.
0.1 N NB 4SCN solution.
Procedure.-l, "Dilute about 4 cc. of Volatile Oil of Mustard, accurately
weighed, with sufficient alcohol to make exactly 100 CC. Transfer 5 CC.
of this solution by means of a pipette to a 100 cc. measuring flask, and add
50 cc. of tenth-normal silver nitrate and 5 cc. of ammonia T.S. Connect
the flask to a reflux condenser and heat it on a water bath for one hour."

The reactions which occur may be represented as follows: First,

the ammonia reacts with the allyl isothiocyanatc with the forma-'"
. tion of thiosinamine:

When the thiosinamine is heated with ammoniacal silver nitrate

solution, it breaks up quantitatively into allyl cyanamide and
hydrogen sulfide:
 ASSAY OF "VOLATILE OILS 383

and the liberated hydrogen sulfide combines with the silver

oxide to form silver sulfide:
2AgNO a + 2NH40H~Ag20 + 2NH 4NO a + H 0 2

Ag 20 + H2S~Ag2S + H 20
The complete reaction may, therefore, be rcpresented as follows:
CaH6NSC + 3NH a + 2AgNOa~Ag2S +
CNNHCaHr; + 2NH 4NO a
2. "Allow the liquid to cool to room temperature, disconnect the fla;;k
from the condenser, add sufficient distilled water to make the mixture meas-
ure 100 cc., mix well, and filter through a filter which has not been previously
moistened. Reject the first 10 cc. of filtrate. To 50 cc. of the subsequent
filtrate, accurately measured, add about 5 cc. of nitric aeid and 2 cc. of ferric
ammonium sulfate T.8., and titrate the exeess of silver nitrate with tenth-
normal ammoniuln thiocyanate. Each cubic centimet~r of tenth-normal sil-
ver nitrate is equivalent to 0.004956 Gm. of allyl isothiocyanate."

Upon filtration, the insoluble silver sulfide is removed, while

the excess of AgNO a passes into the filtrate. This excess of 0.1
N AgNO a is then determined by titration with 0.1 N NH 4SCN,
using ferric ammonium sulfate as the indicator. The reactions
which take place are:
AgNO a + NH4SCN~AgSCN + NH 4NO a
2FeNH 4(S04)2 + 6NH4SCN~2Fe(SCN)a + 4(NH 4)2S04
Each cubic centimeter of 0.1 N AgNO a consumed in the reac-
tion with the hydrogen sulfide is equivalent to 10 X 9;';;0 X 2
0.004947 Gm. allyl isothiocyanate, where 99.13 is the molecular
weight of the latter and 2 is its hydrogen equivalent. In calculat-
ing the percentage of allyl isothiocyanate in the sample, it should
be noted that an aliquot portion of the filtrate corresponding to
one-half of the sample taken is represented in the titration.
384 QUAN'l'I'l'A1'IVE PHAllMACEU'l'ICAL CHEMISTRY

Questions and Problems

1. Why is nitric acid used in the tit.ration of the excess AgNO a with
KONS?
2. Explain each step of the procpdurc giyen in the U.S.P. for the assay
of black mustard.
3. Write equations for all of the reactions that. t.ake place in problem 2.

Assay for Volatile Oil in Spirits.-The estimation of thc volatile

oil content of the certain official spirits is based upon the ~epara
tion of the volatile oil by means of an iml11isc:ible solvent and
measurement of the volume of the oil as illustrated in the follow-
ing exercise.
Exercise 113
Object.-Assay of Spirit of Peppermint.
Materials Required.-5 ee. of spirit of peppermint.
A Babcock bottle and centrifuge.
A 1 cc. pipette.
1 cc. of kerosene.
50 ce. of a saturated solution of calcium chloride (about 30 Gm. in suffi-
cient water to make 50 cc.) acidulated with HCl.
Procedure.-l. "Transfer exactly 5 cc. of the Spirit to a Babcock bottle,
graduated to 8 per ccnt, add exactly 1 cc. of kerosene from [I pipette c[lli-
brated to deliver that amount, and mix well. Then add sufficient saturated
calcium chloride solution, acidified with hydrochloric acid, to almost fill the
bulb of the flask. Rotate the flask vigorously to insure
thorough mixing, then add sufficient of the calcium chloride
solution to bring the separated oil into the neck of the
flask."

The acidified calcium chloride solution, when

added to the alcoholic spirit, throws the volatile oil
out of solution and the oil dissolves in the kerosene
FIG. 64.- when the mixture is shaken. The calcium chloride
Babcock bot-
tIc. (Cour- serves to "salt out" any volatile oil from the diluted
te8y Fisher alcoholic mixture. The acid insures the absence of
Scientific
Company.) alkali which would render acids, traces of aldehydes,
etc., water-soluble. The determination is carried
out in a Babcock bottle (Fig. 64), the neck of which is. calibrated
into eight main divisions of 0.2 cc. each.
2. "Centrifuge for five minutes at about 1500 revolutions per mInute
and then read the volume of oil in the stem. Subtract 5 divisions for the
 ASSAY OF VOLATILE OILS 385
kerosene added and lllultiply the remaining number of divisions by 4.2 to
obtain the volume of oil of peppermint in 100 ce. of the Spirit."

The oil-kerosene layer being lighter than the aqueous layet

rises into the neck of the flask when the mixture is centrifuged.
The length of the oil-kerosene layer supplies the information
needed for the calculation of the per cent of oil in the spirit,
e.g., if 7.4 divisions represents the length of the column of oil-
kerosene in the graduated neek of the flask, the oil measures
7.4 - 5.0 = 2.4 divisions because each 5 divisions are equal to
1 cc. (5 X 0.2 ce.) and 1 ec. of kerosene was added. The 2.4
divisions each of which represent 0.2 ec. are equal to 0.48 CC.
of oil. Since a 5 cc. sample is employed, 100 ec. of the spirit
yielded 0.48 X 20 = 9.60 ec. of oil. The procedure is subject
to slight errol' which has been experimentally determined.
Consequently, the volume of oil in scale divisions is multiplied
by 4.2 (20 X 0.21 cc.) to correct for the error due to solubility
of the oil-kerosene layer in the aqueous layer and also due
to contraction of volume of the immiscible solvents when mixed.
Thus in the example given, 2.4 X 4.2 = 10.08 ec. of oil in 100 cc.
of the spirit.
Similar assay procedures are applied to a number of the official
spirits in the U. S. Pharmacopoeia as indicated in the following
table:
TABLE LIX.-OFFICIAL SPIRITS ASSAYED FOR VOLATILE Ou, CONTEN'r

Amount Official
Spirit used, Fadol' requirement,
ce. per cent
---- ,---
Anise ............................ 5 4.2 9 to 11 V /V
Cinnamon ........................ 5 4.2 9 to 11 V/V
Lavender ......................... 10 2.2 4to 6V/V
Orange, compound .. '., ............. 2 10.5 25 to 30V/V
Peppermint .......... , ............ 5 4.2 9 to 11 V/V
Spearmint ........................ 5 4.2 9 to llV/V
 CHAPTER XXIV

ALKALOIDAL ASSAYING
Proximate Assays.-Alkaloidal assays are commonly referred
to as proximate assays. When such substances as the alkaloids
and glycosides were first isolated from vegetable matter, they
were regarded as plant principles which still retained their
vegetable character, which entered immediately into the composi-
tion of the plant, and which had not been altered in composition.
Consequently, they were called proximate principles as opposed
to the ttltimate principles, such as water, carbon, acetic acid, and
methyl alcohol which had previously been obtained more or less
from all plant products by the methods of pyroanalysis (destruc-
tive distillation).
General Principles.-The alkaloidal drugs and preparations
derived from them constitute a relatively large proportion of the
official substances which are employed frequently in modern
therapy. As a class of medicinal agents, they are characterized
by their high potency. A slight deficiency of alkaloid in a
preparation may cause a lllarked decrease in physiological
effect; on the other hand, a slight excess lllay cause toxic effects
when the preparation is administered. It therefore follows that
the accurate estimation of the quantity of alkaloids present in a
medicinal substance is an important subdivision of pharma-
ceutical analysis.
The assay of alkaloidal drugs and preparations is generally
performed for purposes of standardization, proof of purity,
commercial evaluation, 01' pharmaco-legal purposes. Methods of
various types have been developed for the quantitative estimation
of these principles, e.g., gravimetric, volumetric, colorimetrie,
potentiometric, and physiological. The official assays are limited
to the gravimetric, volumetric, and physiological methods, only
the first two of which come within the scope of this work.
The amounts of alkaloids which occur in cl'ude drugs are subject
to considerable variation in different samples 6f the same drug.
386
 ALKALOIDAL ASSAYING 387

Thc variations may be caused by several factors, i.e.: (1) the age
of the plant when it is collected; (2) the season of the year when
the drug is harvested; (3) the soil and climate in which the drug is
grown; (4) the conditions under which the drug is collected,
dried, and stored. The quantity of alkaloid present in galenical
preparations is also subject to variation due to a number of
factors, some of which are: (a) the quality of drlJ.g employed;
(b) the menstruum used in the extraction of the alkaloid; (c)
the amount of decomposition of the alkaloid during the process
of extraction and during the period of storage.
Many alkaloidal preparations deteriorate comparatively
rapidly, the rate of deterioration being markedly affected by the
nature of the alkaloid, the pH value of the preparation, heat, and
light. Frequent restandardization of the alkaloidal drugs and
their preparations is therefore essential.
In view of the fact that the alkaloids may comprise only a frac-
tion of 1 per cent of the substance assayed and that this small
amount must be separated from numerous other constituents
present in the crude drug or preparation, such as resins, volatile
and fatty oils, coloring matter, glycosides, fatty acids, gums, and
proteins, it is evident that the exact technique involved in any
given method must be carefully adhered to in order to estimate
the variations in alkaloidal content. It is this special technique
that characterizes the chemical assay of alkaloidal drugs rather
than the gravimetric or volumetric nature of the procedure
employed.
The principles employed in the quantitative determination of
the alkaloids by chemical methods are based upon certain
characteristic properties of these substances. The following
general properties are possessed by most of the.members of this
class of compounds: (1) Alkaloids are usually sparingly soluble
in water but readily soluble in certain organic solvents which are
immiscible with water, such as chloroform, ether, amyl alcohol,
benzene, petroleum benzin, or mixtures of these solvents. (2)
Alkaloids combine directly with acids to form salts which are
usually soluble in water but insoluble in certain organic solvents
such as chloroform and ether. (3) Alkaloids are, liberated and
usually precipitated from aqueous solutions of their salts by
alkalies. (4) Alkaloids form highly insoluble precipitates with a
388 QUAN'l'I'l'Al'IVE Pl-IARMACEU'l'ICAL CHE1IH8Tny

considerable number of reagents, especially with the salts of some

of the heavy metals such as mercury, gold, and platinum.
In general, the methods employed to separate the alkaloids
from the other constituents present in a drug involve the use of
the so-called immiscible solvents such as water with ether, water
with chloroform, or water with ether-chluroform mixture. The
process is generally carried out as follows, the separation of the
immiscible solvents being accomplished by means ofa separatory
funnel (Fig. 65): An accurately weighed portion of the drug is
treated with a suitable alkali, whereupon the alkaloids, which
usually occur in the drug in combination with acids as salts,
are set free. An organic solvent such as ether is
~ then added, which dissolves the free alkaloids. An
aliquot portion or all of the ethereal solution
:?:Y"':';:i: corresponding to a definite weight of the sample
::~ ~ taken is separated and shaken successively with
'1 J'
t~ l several portions of acidulated watcr. The complete
extraction of the alkaloids from the ethereal solution
is determined by testing a portion of the latter with
a suitable alkaloidal reagent. Tho alkaloidal salts
formed with the acid dissolve in the water, leaving
most of the resins, coloring matter, fatty substances,
etc., in the ethereal portion. The acid extractions
FIG. 65.- obtained are combined, made alkaline, and shaken
~~~~re~~ory with successive portions of ether. The alkaloids
liberated from their salts by th~ alkali pass into
solution in the ether, leaving water-soluble impurities behind.
The separated ethereal extractives may then be combined,
:waporated to dryness, and weighedj or the residual alkaloid
may be dissolved in an excess of acid of known normality,
and the excess of aeid may be determined by titration with a
standard solution of alkali, using a suitable indicator to show
when the end point is reached.
When the above general procedures are applied in assaying,
numerous difficulties may be encountered which will lead to
serious error unless proper precautions ai'e observed, e.g.:
1. The orga;nic solvents employed are sometimes appreciably
soluble in waterj thus ether is soluble in twelve times its volume
of water at 25C. Consequently, when an organic solvent
 ALKALOIDAL ASSAYING 389

partially soluble in water, such as ether, is employed to extract

the alkaloid from an aqueous solution, some of the alkaloid will
remain in the aqueous solution. This difficulty may be overcome
by extraction with small snecessivc portions of the immiscible
solvent.
2. Resins, fats, and lllany other organic substances remain in
solution in the organic solvent if the aqueous liquid employed in
the shaking-out process is acid, but they dissolve in the aqueous
liquid if it is alkaline, forming soaps, resinates, ctc. Care should
be exercised, therefore, to have sufficient acid in the aqueous
solution used to react with the alkaloid from the organic solvent
and also to neutralize all of the alkali and leave a slight excess of
acid.
3. 'When the liquids are shaken in the separatory funnel,
emulsificatiOil may occm between the organic solvent alid the
aqueous solution, making it exceedingly difficult or impossible to
separate the liquids quantitatively. The formation of emulsions
may frequently be avoided by rotating the separatory funnel
containing the liquids ::;lowly and occasionally inverting it in
place of shaking vigorously. Since the rapidity with which the
alkaloids are extracted from a liquid in which they are insoluble
by an immiscible liquid in whieh they are soluble is largely
dependent on the rate of change of the intcrsurfaee between the
liquids, it is necessary that gentle agitation be continued longer
than is required when strong agitation is employed. If emulsifi-
cation occurs, it is often possible to break the emulsion by some
one or more of the following procedures: (a) addition of more of
the aqueous or organic solvent; (b) gentle stirring with a glass rod;
(c) drawing off the separated solvent as rapidly as it separates j
(d) addition of a few drops of alcohol; (e) addition of a few cubic
centimeters of salt solution i (f) if alkaline, mak.e aeid; (g) if acid,
make alkaline; (h) filtration through a pledget of cotton; (i)
drawing off the emulsion and treating it with anhydrous sodium
sulfate. If none of these methods suffices to break the emulsion,
it may be necessary to start the assay over. Time is always
saved by taking every precaution to prevent the formation of
emulsions. Emulsions formed in the assay of drugs containing
a large proportion of fatty matter should be treated as
follows:
390 QUANTITA'l'IVE PHARMACEUTICAL CHEMISTRY

"Add sufficient N sulfuric add to assure acidity, and evaporate

,the volatile solvent, while stirring with a rubber-tipped glass rod.
When the resinous and fatty matter has been agglutinated, cool
the acid. solution and filter it through a small, wetted filter into a.
separator. Redissolve the residue in 15 cc. of ether, add 5 cc. of
0.1 N acid, evaporate the ether as before, with continued stirring,
and pour the acid solution through the filter into the separator.
Repeat the extraction of the fatty residue with dilute acid two or
three times and finally wash the filter free from alkaloids."
4. When an acid solution contained in a separatory funnel to
which ether has been added is made alkaline, sufficient heat is
frequently generated to volatilize some of the ether and cause
considerable pressure in the funnel. Often the pressure generated
is suffieient to force the stopper out of the funnel with loss
of some of its contents, particularly when the funnel is shaken.
For this reason, it is advisable to permit the mixture to stand and
cool for fI, short time after adding the alkali and to agitate it
gently at first.
Sources of Error.-Numerous possibilities of error are encoun~
tered in alkaloidal assaying other than those common to all
analytical work, such as those caused by inaccurate weighing,
faulty calibration and reading of measuring apparatus, incorrect
strength of volumetric solutions, dirty apparatus, etc. Some of
the more common errors are caused by: (1) failure to secure
complete extraction of the alkaloids fmID the drug or from solu~
tion; (2) loss of volatile solvent during maceration of the drug
before the aliquot portion is taken; (3) imperfect separation of the
immiscible liquidsi (4) decomposition of the alkaloids, especially
those which have an ester structure, upon prolonged contact with
strong acids and alkalies; (5) the use of the wrong indicator.
Other sources of.error, such as variation in the moisture content
of the sample, frequently cause different analysts to obtain non~
concordant results in the assay of the same drug. The most
common cause of variations in results obtained by different work~
ers, however, is referable directly to a lack of familiarity with
the procedures involved, since considerable experience in making
assays of this type is essential to acquire proficiency.
Theory of Distribution Coefficient.-According to Nernst's
law, if two practically immiscible solvents are in contact and 9,
 ALKALOIDAL ASSAYING 391
substance which is soluble in both liquids is added to them, the
substance will distribute itself between the liquids in such a
way that the ratio of the concentrations of the two solutions is a
constant irrespective of the quantity of solute. This constant is
variously termed the distribution ratio, distribuMon constant,
distribution coefficient, or partition coefficient.
If 0 1 is used to designate the concentration of an alkaloid in
ether, and O2 to designate the concentration of the same alkaloid
in an equal volume of an aqueous acid solution, Nernst's law may
be formulated O2 /01 = K, where K is the distribution constant
or coefficient. If there are several solutes snch as alkaloids,
resins, etc., present in the ether when it is shaken with the t),cid
solution, the distribution of each solute is the same as if it were
present alone.
Measurable deviations from Nernst's law take place in alkaloi-
dal assaying due to molecular association and dissociation,
ionization, chemical reaction, etc., but the fundamental principle
which it expresses may be used to explain why it is better to
extract the alkaloid from a solvent with several small portions
instead of one large portion of an immiscible solvent; e,g., assume
that 1 Gm. of atropine in 100 cc. of ether solution when shaken
with an equal volume of dilute sulfuric acid distributes itself so
that 0.1 Gm. remains in the ether layer and 0.9 Gm. in the acid
layer of the solvents, O.Cld/O.tI..r = 9 (the distribution coeffi-
cient). Obviously, only 90 per cent of the atropine present would
be extracted from the ethereal solut.ion by extracting it once with
100 cc. of dilute sulfuric acid solution. If the extraction is
performed with 200 ce. of the acid solution, the a;mount of atro-
pine extracted should be X /200 = 9 and x = 0,947 Gm. atropine.
1 -x
100
Where x = the amount of atropine extracted from the ether
by the acid solution, GOOld = :00' and G.ther = 11~O x. If the
, original solution is extracted with several small portions of acid
solution, i.e., 50 cc., the amount of alkaloid extracted will be
x 1 - x . x/50
G. old = 50 and G.ther = 100 and smce 1 _ x' = 9, x = 0.818
100
Gm. atropine removed by the first extraction with acid, leaving
392 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

0.182 Gm. in the ethereal layer. A second extraetion with a

WI'11 remove 0.182
50 ee. portion of acid soIutlOn _ Xl = 9 ane1
.xI!.50

100
Xl = 0.1489 Gm. atropine where Xl equals the amount of atropine
which passes into the acid solution. Thus, in two extractions,
0.818 + 0.1489 := 0.9669 Gm. atropine are removed from the
ethereal solution by 100 ce. of acid solution, whereas a single
extraction with 200 ee. of acid solution removed only 0.947 Gm.
of thc alkaloid.
Since economy in the use of solvents is an important considera-
tion and large volumes are difficult to handle in shaking out the
alkaloids, it is morc practical to use 'several sUlall portions of
immiscible solvent than one large portion. In the official assay
proceSSOR, 10 to 20 ce. portions of the immiseible solvent are used,
because it has been found that thcse quantitieR serve best to
extract the alkaloids quickly and economically. .'
Choice of Indicators for Alkaloidal Titrations.-In the official'
alkaloidal assays by volumetric methods, the usc of methyl red or
cochineal indicator solution is recommended in all of the titra-
tions. The Rame lot of indicator used in the .standarrnzation of
the volumetric solutions should be employed iil titrating the
alkaloids. Numerous investigators have found that in some cases
other indicators than those employed in the official assays give
more accurate results. The an companying table, page 393,
showing the indicators best suited to the titration of some of
the more hnportant alkaloids, is taken with slight modification
from a paper by H. Wales. 1
Materials Required.-In the statement of the materials
required under the following respective exercises, it is assumed
that the student's locker is supplied with foul' separatory funnels
necessary to conduet alkaloidal assays in duplicate, as well as
measuring cylinders, burettes, flasks, etc. It is also assumed
that the following substances are available in the laboratory:
sulfuric acid, hydrochloric acid, ammonia water, distilled water,
alcohol, mercuric potassium iodide test solution, iodine test
solution, and methyl red and cochineal indicator solutions. The
quantities of immiscible organic solvent required are approximate.
1 WALES, n., Jour. Ind. Eno. Chem., 1926, 18, 390.
 ALKALOIiHL ASSAYING 39S
TABLE LX.-INDlCA'l'OnS ~'on ALKALOIDAL TITRATIONS

pH range
Salt of Average for Indicator
pH
indicator

Aconitine ................. . 5.04 4.2 to 5.8 Methyl red

Arecoline ................. . 4.81 3.8 to 5.8 Methyl red
Atropine ................. . 5.56 3.8 to 7.2 Methyll'ed
Brucine .................. , 4.85 3.9 to 6.0 Methyl red
Cephaeline ......... , ..... , . 4.81 4.2 to 5.4 Methyl red
Cinchonine .. , ..... , , , .... . (L02 5.5 to 6.5 Bromcresol purple
;:Cinchonidine ...... , .. , , ... . 5.90 5. 4 to 6.4 Bromcresol purple
Cocaine .. , ............... . 5.20 4.0 to 6.5 Methyll'ecl
Codeine., ................ . 4.86 3.6 to 6.3 Methyll'ecl
Cotarnine ..... , ......... . 5.97 4.9 to 7.0 Bromcresol purple
Di:n:cetyl morphinc ......... . 4.89 4.2 to 5.7 Methyl red
Enletine .................. . 4.90 4.2 to 5.6 Methyl red
Ethyl hydrocupreine ....... . 6.33 5 . 6 to 7.0 Bromcresol purple
Ethylmorphine ........... . 4.99 4.2 to 5.8 Methyl red
Homatropine .............. . 5.74 3.9 to 7.6 Methyl red, propyl reel,
bromcresol purple
Hydrastine ............... . 4.45 3.8to 5.0 No eud point with
methyl red or brolll~
phenol blue
Hyoscine ................. . 4.83 3.6 to 5.6 Bromphenol blue
Ityoscyamine ............. . 5.83 3.8 to 7.6 Methyl red, propyl red,
bromcresol purple
Morphine ......... , ....... . 4.68 4.0 to 5.2 Methyl red
Narcotine .............. , .. 4.43 3.9 to 4.9 Bromphenol blue
Nicotine ............. , .... . 5.26 4.4 to 6.1 Methyll'ed
Papaverine. , ............. . 4.23 3.8 to 4.6 Indistinct, cannot be
titratecl
Phsostigmine ............ . 4.85 3.8 to 6.0 Methyl red
Pilocarpine ............... . 4.31 3.6 to 5.0 Bromphenol blue
Qiiinine .................. . 6.12 5 . 5 to 6.5 Bromcresol purple
Quinidine ................. . 6.10 5 . 5 to 6.5 Bromcresol purple
Strychnine. , .............. . 4.81 3.8 to 6.0 Methyl red
Thebaine ................. . 5.08 4.0 to 6.2 Methyll'ed
Yohimbine ................ '1 4.72 4.0 to 5.3 Methyl red

Alkaloidal Test Solutions.-A number of reagents form insol-

uble compounds with many of the alkaloids precipitating the
latter from aqueous solution. In many cases, the presence of a
mere trace of alkaloid can be detected by adding a suitable
394 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

reagent to its solution. Some of the frequently used reagents are

auric chloride, platinic chloride, mercuric chloride, gallotannic
acid, picric acid, phosphomolybdic acid, potassium mercuric
iodide, and iodine in potassium iodide. While most of these
reagents are employed in qualitative tests for the alkaloids,
only the last two are of importance in the official assays, since they
are employed to test for the complete extraction of the alkaloids
by immiscible solvPnts.
Mercuric potassium iodide test solution, commonly known as
Mayer's reagent, is prepared by dissolving 1.358 Gm. of mercuric
chloride in 60 cc. of distilled water and 5 Gm. of potassium iodide
in 10 cc. of distilled water, mixing the two solutions and diluting
the product to 100 ce. with distilled water. Mayer's reagent
forms curdy yellowish or white precipitates with minute traces
of many of the alkaloids when it is' added to their acidulated
aqueous solutions. The composition of the precipitates usually
varies somewhat depending upon the conditions under which the
precipitation is conducted. Mercuric potassium iodide is a very
sensitive reagent for strychnine and quinine; it gives a much less
delicate reaction with atropine, coniine, and morphine; and col-
chicine, caffeine, and theobromine are not precipitated by it at
all.
Iodine test solution, commonly known as Wagner's reagent, is a
solution containing approximately 1.2692 Gm. of iodine and 1.8
Gm. of potassium iodide dissolved in sufficient distilled water to
make 100 cc.; 0.1 N iodine solution may be employed. Wagner's
reagent yields reddish or red-brown precipitates of complex
composition with nearly all of the alkaloids, even in dilute solu-
tions, when it is added to an aqueous solution slightly acidified
with sulfuric acid. The precipitates are usually amorphous.
The qi.l!1ntity of the reagent used should not be sufficient to color.
the solution more than a faint yellow.
General Procedures. 1.. Selection of the Sample.-The sample
of a crude drug taken for assay must represent the entire drug
and be powdered to a specified fineness. Usually the drug must
be ground to a fine powder (No. 60), but finer powders may be
employed. When the whole orude drug such as leaves or
seeds is powdered for assay, care must be exercised not to
exclude leaf stems, seed coats, etc., since the alkaloid may be
 ALKALOIDAL ASSAYING 395
localized in amounts varying in the different mo~phological parts
of the drug. Care should be bken to avoid the loss of watp.l'
during the powdering of the drug. If it is impossible to avoid this
loss, the drug should be dried at a low temperature before
powdering, the loss of water noted, and a correction made in the
final calculations.
2. Weighing of Portions for Assay.-In the preparation of
drugs for assay, all portions directed to be weighed should be
weighed accurately, but with crude drugs, accuracy to the second
decimal point is sufficient. Portions of pilular extracts may be
weighed on a piece of waxed or parchmentizcd paper, the surplus
paper cut away, and the extract and adhering paper dropped into
a separator, beaker, or dish containing the solvent, and the
extract dissolved. In transferring weighed portions to a sepa-
rator, the dish containing the material to be assayed should be
thoroughly rinsed and the rinsings added to the separntor.
3. Methods of Extraction.-The alkaloids are extracted from
drugs by the process of maceration, by the process of percolation,
and by the process of continuous extraction. The maceration
process is adapted to those drugs which contain a relatively
large amount of alkaloids and which, because of their nature or
the nature of the drug, are difficult to extract completely by
the process of percolation. In the maceration process, it is
assumed that all of the alkaloids are diRsolved in the organic
solvent and that an aliquot part of the solvent contains an
amount of the extracted alkaloids in definite proportion to
the weight of drug taken as sample. Many analysts believe that
the aliquot portion of solvent obtained by the maceration
process does not contain exactly the assumed proportion of the
total alkaloids. Consequently, methods for the total extraction
of the alkaloids are regarded as more satisfactory.
A. By Maeeration.-An accurately weighed portion of the
ground drug is treated with the specified solvent or mixture of
solvents made alkaline with ammonia T.S. and thoroughly
mixed and allowed to macerate for from 12 to 24 hr. with occa-
sional agitation, or for a shorter period with continuous agitation.
At the end of this period, the drug is allowed to settle and then
an aliquot portion of the solvent is decanted and used for the
extraction of alkaloids.
396 QUANTI'l'Jl'l'IYE PHARMA_CEUTICAL CIIEMIHTRY

The organic solvent, accurately 111.casured from a calibrated

cylindrical graduate, is added and allowed to stand in contact
with the powdered drug so that the small partides of the drug
will be impregnated with the organic solvent before the ammonia
is added. If the ammonia test solution is added first, the drug
particles become wetted by the water with which the organic
solvent is immiscible, and the latter will not permeate the drug
cells to extmct the alkaloids completely. The alkaloids generally
occur in combination ",-ith various acids as salts in the crude drug.
The ammonia reacts .vith these alkaloidal Rnlts, forming the free
alkaloids and the ammoniulll salts of the acids, and the liberated
alkaloids dissolve in the organic solvent. The process of extrac-
tion of the alkaloid from the cellular structure of the drug is
probably of an osmotic nature which proceeds slowly. Con-
sequently, a period of maceration is required to permit thc com-
plete transference of the alkaloids from the drug cells to the
organic solvent. The rate of extraction is accelerated by grinding
the drug to a fine powder, thereby breaking down its cellular
structure, and by frequent agitation of the drug-solvent mixture
so that the solvent in eontnct with the particles is ehanged
frequently, The small particles of drug which sometimes remain
in suspension in the organic solvent become hydrated when the
latter is shaken with a little water and settle out. The flask in
which the maceration and agitation of the drug with the sol-
vents are carried out should be tightly stoppered throughout the
entire period of maceration. to prevent the escape of volatile
organic solvent, since the volatilization of some of the solvent
might cause aeonsiderable error; e.g., if 100 co. of solvent is taken
for the extraction of the alkaloids from a drug and 10 cc. is,
lost by volatilization, and an aliquot portion of 50 cc. representing _
one-half of the weight of the sample is measured for assay, the
aliquot portion will contain five-ninths instead of one-half of the
alkaloids contained in the drug, and an error of approximately"
5 per cent will be caused in the assay result.
An aliquot portion of the supernatant organic solvent is
poured off from the mixture and measured accurately in a cali-
brated cylinder; e.g., if 4 Gm. of a drug is macerated with 100 cc.
of solvent and a 50 cc, portion of the solvent, is decanted ,_the
50 cc. portion of solvent will contain an amount of extractive
 ALl(ALOIDAL ASSAYING 397
equivalent to 5~i 00 X 4 = 2 Om. of the drug, provided that none
of the solveI)t originally added is lost by volatilizatioIl. The
aliquot portion of solvent should be mcaflurecl at the same tem-
perature at which the original solvent was measured, since the
volume of liquids varies somewhat with changes in temperaturt'.
B. By Percolation.-An accurately weighed quantity of the
ground drug is placed ill a suitable container and completely
saturated with the flpecied solvent or mixture of solvents and
allowed to stand for 5 min. A sufficient quantity of ammonirL
T.S. to make the drug alkaline is adclpd and thoroughly ll1ix(~d
with the drug. The moistened drug is transferred to a cylindrical
percolator, previously prepared by packing the outlet with
purified ootton. A small amount of the solvent may be used to
rinse the container and the rinsing added to the percolator.
The drug is allowed to macerate for a suitable period of time (from
1 to 12 hr. or over night, depending upon the drug to be assayed),
then the drug is firmly packed, a pledget of purified cotton plaeed
above it, and percolated slowly until the drug is eompletely
extracted. Complete extraction of the alkaloid is determined by
evaporating about 4 cc. of the last pereolate to dryness, dissolving
the residue in dilute acid, and adding a drop of nwrcnric potns-
sium iodide T.S. or, whcn testing for caffeine or eolehicille, a drop
of iodille T.S. There should be no turbidity produced by these
reagents. The percolate is then treated for the extraction of the
alkaloids.
The extraction of the alkaloids from a drug by cold percolation
has the disadvantage that it requires it relatively brge amount
of solvent to effect con1plete extraction. The cold percolation
process of extraction is bcst suited to the extraction of drugs
which contain unstable alkaloids or which yield a large amount
of l1011-alkaloicltll extractive by hot percolittion. The cold
percolation proceSR is well suited to the extraction of those drugs
which contain a comparatively small percentage of alkaloids
and which can be readily extracted by percolation. The impor-
tance of thetotal extraction of the alkaloid from a drug such as
hyoscyamus becomes apparent when it is recalled that the official
drug is required to contain only 0.040 pel' oent of alkaloids.
The percolator employed in the extraction of the drug shOUld
be of It special type similar to that illustrated in Fig. 66. The
398 QUANTiTATIVE PHARMACEUTICAL CHEMIS'l'RY

barrel of the percolator should be about 15 or 16 cm. long and

about 3 or 4 cm. in diameter., A percolator of these dimensions
will accommodate from 10 to 25 Gm. of drug, and, because of
the small diameter, a long column of the drug will be provided
through which the menstruum must pass. The glass stopcock
is necessary to eliminate the use of rubber connections, since
rubber is dissolved to some extent by chloroform and ether.
The alkaloids are liberated by the ammonia and they dissolve
in the organic solvent as described under A.
C. By Continuous Extraction.-An accurately
weighed portion of the ground drug is placed in
an extraction thimble and the thimble trans-
ferred to a ~uitable extractor (a Soxhlet extractor
of appropriate size is satisfactory). The drug is
i! moistened with the specified solvent and mixed

1 y~
.
; 32 :
i

L18~
"
'i 2
..
- , - . \ . /~'6

10
0

"
by means of a stirring rod and allowed to stand
about 5 min. It is then made alkaline with
the specified quantity of ammonia T.8. and
thoroughly mixed. The stirring rod is rinsed
. with a small portion of the solvent and the drug

-" 'ii' macerated for from 6 to 12 hr. or over night.

L.o. . The drug is covered with a pledget of purified
FIG. 66.-Per-' cotton and packed in the thimble, a sufficient
(lola tor recom- quantity of solvent is added and the drug
mended for type
process B (me,,~- extracted for a specified period of time. The
~re~e,:,ts are given solvent remaining in the extraction chamber is
10 mIllimeters) .
transferred to the receiving flask and the liquid
extract treated for the extraction of alkaloids.
An extraction apparatus of the type illustrated in Fig. 60
and described on page 337 is very satisfactory. The continuous
extraction process requires a small amount of the organic solvent
and, in most cases, complete extraction of the alkaloids is obtained
in from 3 to 5 hr.
4. Extraction of the Alkaloids from Organic Solvents with Acid.-
The alkaloids are extracted from ethereal or chloroformic
solutions by shaking with dilute acids. The concentration and
quantity of the acid employed are left to the discretion of the
analyst, but the use of 10 cc~ of a normal or 5 per cent acid or
sufficient to render the mixture distinctly acid for the first extrac-
 ALKALOIDAL ASSA YING 399
tion and of 10 cc. portions of 0.5 N or 2 per cent acid for succeed-
ing extractions 'will be found to be practical and convenient. The
shaking of the immiscible solvents should not be violent, but it
should be sufficient to mix them thoroughly without emulsifi.ca~
tion. Repeated successive extractions with small portions of thE
acid must be continued until the alkaloid is completely removed
from the ethereal or chloroformic liquid as shown by tests with
the designated reagents. Each portion of the acid solution
should be carefully separated from the organic solvent and run
directly into another separatoryfunnel. In all assays,the extrac-
tion should be continued until 0.5 cc. of the last acid washing
shows no turbidity on the addition of a drop of mercuric
potassium iodide T.S., or, in the case of caffeine and eolchieine,
on the addition of a drop of iodine T.S. If traces of organic
solvent are drawn off with the acid solution, they may be
removed by washing the combined acid extractives with 2 to 3
cc. of ether or chloroform which is separated and washed in tUrn
with about 5 cc. of distilled water, the aqueous washings being
separated and added to the acid solution.
If sulfuric acid is used, the acid converts the free alkaloids dis-
solved in the organic solvent into their sulfates which pass into
solution in the aqueous acid layer of the immiscible solvents when
the mixture is shaken, leaving chlorophyll, resins, fixed and
volatile oils, and other impurities dissolved in the organic solvent.
The rate at which the alkaloids pass from solution in the organic
solvent into the acid solution is increased greatly by shaking the
mixture, since a greater area of contact surfnce between the
liquids is produced. Sulfuric acid solution is employed to extract
the alkaloids, because the ammonium sulfate formed in the next
step in the procedure is not appreciably soluble in the organic
solvent used for thc final extraction of the alkaloid.
5. Extraction of the Alkaloids from Acid Solution with Organic
Solvents.-The acid solution of the alkaloids contained in a
.' separatory funnel is made alkaline, in most cases with ammonia
T.S., and then shaken with several successive portions of organic
solvent, such as ether, chloroform or ether-chlorofol'lll mixture.
The volume of organic solvent used in each extraction should be
at least one-half that of the aqueous solution. The extraetion
with portions of organic solvent must be repeated as long as any
400 QUilNTITATIYE PHL1RMACEUTICL1L CHEMISTRY

alkaloid is extracted by the solvent. To determine the comple-

tion of extraction, evaporate 1 cc. of the last washing and dissolve
the residue in a few drops of diluted hydrochloric acid; the result-
ing solution should show no turbidity on the addition of a drop
of mercuric potassium iodide T.S., or, in the case of caffeine and
eolehicinc"on the !1cldition of a drop of iodine T.S. The number
of extractions required depends largely on the character of the
alkaloid. With most alkaloids it is advisable to extract three or
foUl' times before testing. Physostigmine and pilocarpine
require about twice as many extractions as other alkaloids.
The ammonia reacts with the alkaloidal sulfates contained in
the aqueous solution to form the free alkaloids and ammonium
sulfate. If considerahle quantities of alkaloid are contai,ned in
the acid solutioll, the alkaloids may be precipitated when the
alkali is added. Consequently, the first portion of organic
solvent is added prior to the addition of the ammonia water to
dissolvo any alkaloidal precipitate formed. If this procedure is
not followed, the precipitate frequently agglomerates into large
particles which dissolve slowly when the organic solvent is
added.
6. Evaporation of Organic Solvents.-The evaporation of the
final alkaloidal solution in organic solvents may be performed by
distillation with recovery of the solvent, by evaporation on a
steam bath or sand bath, or in a current of warm air. When the
evaporation is conduded on a steam bath, precautions must be
observed to prevent the ignition of the vaporized solvent. The
operation may be performed by heating a prepared water bath
until the water boils, turning out the flame, and placing the
alkaloidal solution eontained in a suitable container on the bath.
If a sand hath is employed, it should preferably be of the steam-
heated type and the temperature of the bath should be about
60C.
Chloroform. tends to form unstable compounds with many
alkaloids; consequently, when chloroformic solutions of the
alkaloids are evaporated, the residue may retain traces of
chloroform rather tenaciously. These traces of chloroform are
best removed by treating the alkaloidal residue with 2 to 3 cc.
of ether or alcohol and evaporating the solution to. dryness again.
This is a .wise precaution to ,emploY in the evaporation of all
 ALKALOIDAL ASSAYING 401
solutions of alkaloids in organic solvents when the final deter-
mination is made by volumetric methods ill order to remove small
traces of ammonia which lllay be oceluded in thr alkaloidal resi-
due, particularly if the latter is gummy in n:1tnre.
Those alkaloids which have the structure of ('.<;ter8 such us
atropine, hyoscyamine, and cocainp and Henne others are decom-
posed more or less by heat anel by prolonged ('.ontact "ith water
and strong acids and alkalies. Consequently, their solutions
should be evaporated at a low temperature, usually at about
60C.; and, if it is necessary to allow the alkaloid in solution to
stand from one laboratory period to the l1ext before the assay can
be completed/ it should be shaken out of acid or alkali solutions
into an organic solvent and kept in a tightly stoppered flask.
7. Gmvimetl'ic Determinations of Alkaloicis.-If the alkaloid is to
be estimated gravimetrically, the solvent is evaporated in the
usual manner from a previom;iy weighed evaporating dish,
wide-mouth flask, or beaker, and the residue is dried to constant
weight in an electric or hot-air oven and weighed, The same
procedure should be followed in drying the flask or bcakm used
as is employed in drying the container with its residue. When
the residue is to be weighed, if the final solvent has been chloro-
form, the last traceR of that solvent should be removed by
the addition of a little neutral ether or alcohol, followed by
evaporation. Care must be taken to avoid loss by decrepitation,
especially when evaporating chloroformic solutions of nux
vomica or cinchona alkaloids. Decrepitation may usually be
prevented by the addition of a little alcohol after the solution
has been reduced to a volume of 1 or 2 cc., evaporating at a low
temperature, and rotating the container during the process.
8. Volumetric Det61:minations of Alkaloids.-If the alkaloid is
directed to be assayed volumetrically, the residue should be
softened by the addition of about 1 cc. of neutral alcohol 01' ether,
the required amount of standard acid added, and the mixture
gently warmed to insure the cOlllplete solution of the alkaloid.
If preferred, the alkaloidal residue may be dissolved in chloro-
form, the standard acid added, and the chloroform removed by
evaporation. Before titrating add a sufficient quantity of dis,
tilled water to make the volume of the mixture measure about
25 cc. The amount of acid in excess of that which cOlnbines
402 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

with the alkaloid is then determined at once by titration with

standard alkali.
As a rule the volumetric method of determining the quantity
of alkaloid obtained in a given assay is more accurate than the
gravimetric method, since many impurities which cannot be
removed are present in the alkaloidal residue and are computed as
alkaloid when the residue is weighed. If the residue is titrated,
however, only those impurities which are acidic or basic affect
the result.
Adsorbents.-In assaying iluidextracts, tinctures, and other
preparations of alkaloid-bearing drugs, it is often necessary to
evaporate these to dryness arid, to avoid loss and to aid in the
evaporation, they are usually added to somc adsorbent material.
Paper pulp or asbestos fiber should be used for this purpose.
Such adsorbent material must be acid and alkali washed and then
rendered neutral by washing with distilled water and dried before
being used.
Apparatus for Proximate Assays.-When a container of definite
size and shape is recommended in a proximate assay process, it
is understood that this is advisory and not obligatory, except
when volumetric flasks, measuring burettes, or other exact
measuring apparatus are specified.
 CHAPTER XXV

OFFICIAL TYPE METHODS

The official alkaloidal drugs and the galenical preparations
containing alkaloids are assayed by two type methods anc1
by special methods. The type methods are (1) the aliquot part
method for drugs and galenical preparations and (2) the total
extraction method for drugs and galenical preparations. The
procedures in tho following assays are taken almost verbatim from
the U.S.P. and N.F., slight changes being introduced where
necessary to include more specific details.
Gravimetric Assays by the Aliquot-part Method
Exercise 114

Object.-Assayof Hydrastis for Ether-soluble Alkaloids.

Materials Required.-10 Gm. of hydrastis in No. 60 powder.
200 cc. of ether.
Procedure.-l. II Place 10 Gm. of Hydrastis, in fine powder and accurately
weighed, into a suitable container, add 100 cc. of ether, allow the mixture to
stand about five minutes, and add 10 ce. of ammonia water. Shake the
mixture for one hour in a mechanical shaker or during two hours intermit-
tently, and then set it aside overnight. Again shake it during one half hour,
allow the drug to settle, and decant 50 cc. of the clear ethereal solution."
(See General Procedure 3A, page 395.)

The alkaloids found in the rhizomes and roots of hydrastis con-

sist chiefly of hydrastine and berberine in varying proportion.
The latter is almost inactive, while the hydrastine is very active
physiologically. Consequently, a determination of the total
alkaloids of hydrastis would not be a correct Illeasure of the
therapeutic activi.ty of the drug. Ether is used, therefore, as the
organic solvent in the extraction of the alkaloids, because hydras-
tine is readily soluble and berberine is practically insoluble in this
solvent. The aliquot portion of 50 ce. of the ethereal solution is
equivalent to 5 y! 0 0 X weight in grams of the sample of hydrastis.
403
404 (JUANTl1'A'l'IVE PHARMACEU'l'ICAL CHEMISTRY

2. "Extract tJHl allmloids completely from the ethereal solution by shak-

ing with successive small portions of dilute sulfuric acid (about 1 per cent).
Combine the auid solutions in a separator, add about an equal volume of
ether, then add a slight excess of ammonia water, and immediately shake to
extraet the alkaloids." (See General Proeedure 4, page 398.)

The sulfuric acid converts the hydrastine into hydrastine

sulfate which is soluble in the acid solution.
3. "Sepf1l"ate the ethereal layer, and eomplete the extraction of the
Hlklloid~ from the alkaline llqueous layer by shaking with successive portions
of ether." (See General Procedure 5, page 399.)

The hydrastine liberated from its sulfate is practically immluble

in water, but it dissolves in ether to the extent of 10m:. in
175 cc. at 25C. Traces of berberine carried over into the acid
solution are eliminated by using ether as the immiscible solvent,
since berberine is soluble in water (10m. in 4.5 cc. at 22C.)
and insoluble in ether.
4. "Filter the ethereal solutions and evaporate the ether. Dry the resi-
due to constant weight at 100 0 e.
"The weight obtained represents the yield of anhydrous ether-soluble
alkaloids from 5 Gill. of Hydrastis." (See General Procedures 6 and 7,
page 400.)

The percentage of ether-soluble alkaloids contained in the root

is found by siIllple calculation as follows:
wt. of alkaloidal residue X 100 t
0.5 X wt. of hydrastis = per cen .

The Pharmacopoeia requires that hydrastis contain not less

than 2.5 per cent of ether-soluble alkaloids. Compare the per-
centage of alkaloids found in the sample assayed with the official
requirement.
The extract, fiuidextmct, and tincture of hydrastis are assayed
by procedures similar to that employed in the assay of hydrastis.
Questions and Problems
1. Why is hydrastis assayed for its content of ether-soluble alkaloids?
2. Explain in detail how to test for hydrastine in the ethereal solution
which has been shaken out with acid.
S. Can hydrastine be estimated vplumetrically (sec table of indicators,
page 286)1
4. What is an aliquot portion of a solution 1
 OFP'ICIAL TYPE METHODS 405
6. vYhy isit more practical to usc tlUlun!Jcr of sIlIall portions of imllli~cibic
solvent in shaking out the alkaloidH. tlU\!! to use olle or t,\yO lluge portion.,?

Exercise 115
Object.--Assay of Cinchona for Total Alkaloills.
Materials Required.-5 Gm. of cinchona in No. 00 powder.
200 ceo of etlwr-ehlnroform mixture.
100 ce. of chloroform.
Procedure.-l. "Placc 5 GlU. of Cinchoua, in fine powdcr, and 15 (:11. of
3 per cent hydrochloric acid in a 500-ee. flitsk and heat the mixture on !L
water bath for one hour. Cool and add 200 ce. of ether-chloroforrnie solu-
tion (ether, 3 volumes, chloroform, 1 volume) and 10 ee. of stronger tlIlllllollia
T.S. Stopper the flask tight1y lWeI Rhake it, for one hour in a medll1.uil'al
shaker. Allow t.he Inixtt)rc to stand ovcr night, again shake it for one-half
hour, and then allow the drug to settle. (If the supnrnatan't liquid il? not
clear, add a few ce. of dist.illed wl1ter, again shake the contents of the flask
vigorously, and allow the drug to settle.)" (See General Procedure 3_,t,
p!1ge 395.)
Cinchona bark contains about 20 alkaloids the most important
of which are quinine, quinidine, cinchonine, anel eirwhonieline.
The alkaloids occur as salts in combination with quinic and cin-
chotannic acids. These Ralts arf decomposed ",hp11 boiled with
hydroehloric acid ">1th the formation of the corresponding
alkaloidal hydrochlorides which are soluble in the water. 'When
the aqueousRolution is made alkaline with ammollIa, the liberated
alkaloids pass into solution in the ether-chloroform mixture
almost completely, due to the large volume the latter of
employed.
2. "Quickly decant 160 ce. of the clear, ether-chloroformie solution, meas-
ured at approximately the same temperature as the original ether-chloro~
fonnic solution and representing 4 Gm. of drug." (Sec General Procedure 3,
page 395.)
The aliquot portion is equivalent to 16%00 X the weight of
the sample taken.
3. "Transfer the solution to a separator, rinse the measuring vessel with
a small quantity of the original menstruum and add the rinsings to the
Aepamtor. Completoly extract the alkl1loids with llpproximately5 per cent
sulfuric add and collect the acid solution of the alkaloids in a second sepal'U.-
tor." (See Geneml Procedure 4, page 398.) ,

The alkaloidal sulfates are formed and dissolve in the acid

solution.
406 QUANTITATIVE PHARMACEUTICAL CHEMISTRY
,
4. "Make the acid solution strongly alkaline with ammonia T.S. and
completely extract the alkaloids with chloroform. Evaporate or distil the
chloroform in a tared beaker or flask and dry the alkaloidal residue to con-
stant weight at 1000. The weight obtained, multiplied by 25, represents
the per cent of the alkaloids of cinchona in the drug." (See General Pro-
cedures 5, 6, and 7, page 399.)
The ammonia water reacts with the alkaloidal sulfates,
setting the alkaloids free and forming ammonium sulfate.
Only a slight excess of ammonia should be used to make the
solution alkaline, since the cinchona alkaloids are appreciably
soluble in strong ammonia solution. The alkaloids set free by
the alkali dissolve in the chloroform.
It has been reported that the cinchona alkaloids form addition
products with chloroform which are not entirely broken down
when the residue from the ehloroformic solution of the alkaloids
is dried at 100C. The error in the assay results caused by the
retention of chloroform by the alkaloidal residue when the latter
is dried as directed, however, is negligible.
The gravimetric method is employed in the determination
of the alkaloids in the residue, because the indicator end point
is not sharp when the mixture of alkaloids is titrated.
The U.S.P. requires that cinchona contain not less than 5 per
cent total alkaloids when assayed by the above method. Calcu-
late the percentage of alkaloids found in the sample assayed
and compare the results with the official requirement.
Questions and Problems
1. Why is hydrochloric acid used in the initial menstruum to extract the
alkaloids from cinchona?
2. Does the ether-chloroform mixture extract practically all of the alkaloid
present in the sample in the first step?
3. Why should the addition of a large excess of ammonia to the acid
solution of the alkaloids previous to the shaking out with chloroform be
avoided?

Exercise 116
Object.-Assay of Compound Tincture of Cinchona.
Materials Required.-50 ce. of compound tincture of cinchona.
200 cc. of chloroform.
200 cc. of ether.
Procedure.-L "Accurately measure 50 cc. of Compound Tincture of
Cinchona. and evaporate it, at a temperature not exceeding 100C., to a
 OFFICIAL TYPE METHODS 407
volume of about 10 cc. Add sufficient asbestos fiber or paper pulp to
adsorb the liquid, and continue the evaporation to dryness. Transfer the
residue to a flask or bottle, add 200 cc., accurately measured, at room tem-
perature, of ether-chloroform mixture (ether 4 volumes, chloroform 1 vol-
ume) and sufficient ammonia T.S. (which may be used to rinse out the
adhering portions of the Tincture from the evaporating dish) to render the
mixture strongly alkaline. Securely stopper the container and shake it
mechanically during one hour, or intermittently during two hours, and then
allow the mixture to stand over night."
The tincture is evaporated to drive off the alcohol which is
miscible with ether and chloroform. An adsorbent such as
asbestos fiber or paper pulp (filter paper torn into small pieces)
is added so that the non-volatile residual matter consisting
largely of alkaloidal hydrochlorides, glycerin, and cinchona
red will be deposited as a thin film when the tincture is evapo-
rated. If some adsorbent is not used, a resinous, varnish-like
residue usually results from which the alkaloids cannot be
extracted readily. The addition of ammonia T .S. liberates the
alkaloids which dissolve in the ether-chloroform mixture along
with the glycerin, cinchona red, and considerable other extractive
matter.
2. H Again shake the mixture intermittently for half an hour, allow it to
settle, quickly decant 160 cc. (representing 40 cc. of the Tincture) of the
approximately clear liquid. Filter this into a separator and wash the
measuring vessel with sufficient of the ether-chloroform mixture, adding
the rinsings to the filter. Extract the alkaloids froUl the clear Jiquid with
acidulated water, using sufficient dilute sulfuric acid to render the contents
of the separator and each extract distinctly acid to litmus paper. Pass the
acid extracts in succession through a wetted, double filter into It second
separator. Render the combined liquids distinctly alkaline with stronger
ammonia T.S., and extract with chloroform. Pass the chloroformic
extracts through a double filter, which is kept saturated with chloroform,
into a suitable, tared receptacle. Evaporate the chloroform on a water
bath, dry the residue to constant weight at 100C., and weigh. The weight
multiplied by 2.5 indicates the weight of alkaloid~ in 100 cc. of the Com-
pound Tincture of Cinchona." (See Assay of Cinchona, Procedures 2, 3,
and 4, page 405 for explanation.)
Volumetric Assays by the Aliquot-part Method
Exercise 11'1
Object.-Assay of Ipecac for Ether-soluble Alkaloids.
Materials Required.-10 Gm. of ipecac in No. 60 powder.
200 ee. of ether.
408 QUA_N'l'I'lWl'[ VI!] P1!ilRMACEU'l'IC.1L CHEMISTRY

Procedure.-l. "Plnee 10 Gm. of Ipecac in fine powder in a dry, 250-ce.

flask. Add 100 eo. of ethel', which is free from peroxide, stopper the flask,
shake the mixture thoroughly, and allow it to stand for five minutes; then
add 10 ce. of ammollia T.S. Agnin stoppel' the flailk tightly and shake it for
one honr in a Illeehanical shaker, 01' illtermittently during two hours.
Allow the mixture to st!md over night, again shake it intermittently Juring
. olle-h:nlf hOllr and then fdlow the clmg to settle. Decant into a Heparator
50 cc., representing 5 Gill. of Ipecae, aeclll'l1tely measured, of the clear,
supernatant liquid, rinsing the measuring vessel with a RmaH quantity of
ethel' and adding the rinsings to the solntion in the separator." (See Gell-
eml Procedure 3A, piLge 395.)

Ipecac contains a number of alkaloids, the most important of

which [He ellietine,. cephaeline, and psychotrine. These three
alkaloids occur in varying' proportions in different varieties of
the root. Psychotrine constitutes a relatively small proportion
of the total alkaloids and is not known to be of importance
therapeutically. Consequently, this alkaloid is eliminated in
the assay process, since it, is practically insoluble in ether, while
the liberated emetinp, and cephaeline, being soluble in ether,
are determined. The ipecac must be finely powdered, since it
is not penetrated readily by the solvent duc to its high starch
content.
The aliquot portion of the ethereal solution contains an
amount of emetine and cephaeline equivalent to one-half the
weight of sample taken.
2. "Completely extract the alkaloids from this ethereal solution with
approximately normal sulfuric acid, preferably using 15 cc. the first time
and 10 ce. on each succeeding extraction, and filtering IOlaeh portion into a
second sePllrator. Continlle the extractiOll until no lellctioll can be detected
in the sulfuric' acid 'solution 'when tested." (See General Procedure 4,
p1\:ge 398.)
,.
" The sulfates of emetine and cephadjllf~ are formed and dissolve
in' the aqueous solution.
3. "To the combined acid solutions add about an equal volume of per-
oxide-free ether, render the mixtnre alkaline by the addition of arnmonin
T.S., nnd extmct with'sliccessive portions of the ether until no -Visible renc-
tion takes place when tested as directed apove. Filter each portion of the
ethereal extract into a 200-cc. finsk or' beaker, and cnrefully evaporate the
combined ethereal solut.ions on a steam bath until nearly but not quite dry.
Add 5 cc. of the peroxide-free ether and again evaporate nearly to drynel:!s.
Add 10 ce. of tenth-normal sulfuric acid and heat on a stenm bath to effect
 OFFICl!1L TYPE METHODS 409
complcte solution tllld to rcmoye all of the ether. Cool, and titrate t11['
excess of acid with tenth-normal sodium hydroxide, using methyl rpd T.S.
as the indicator. Each cubic centil(loter of tenth-normal sulfllric acid is
I.lquivalent to 0.0240 Gm. of the et)lil~-soluble alkaloids of 11)Ccao." (See
General Proecdutes 5, 6, and 7, page 399.)

The ammonia sets the emetine, ,and cephaeline free;' and the
liberated alkaloids dissolve in the ether.
The residue cOllsi!?ts almost entirely ofcnlPtine and cephaeline
which dissolve in the standard aeid solution,. forming emetine
and cephaeline sulfates, as illustrated by the following reaction
for emetine:
C2uH4004N 2 + H2S0C--.?C20H4004N 2.H 2SO{
Each cubic centimeter of 0) N H 2 SO,1 consumed corresponds to
0.024 Gm., of the ether-:soiub~9 alkaloids,of, ipecac.
The Ph8;rm?-copo~~a xequ~J;gs that ,ipecac, contain not leSH than
1.75 per cent of ether-soluble l1Ikaloids" Calculate,the percentage
of ether-soluble alkaloids contained in the sample assayed and
compare the results with ~he,official requirement. .
Fluide::dract of Ipecac, U.S.P., and Tincture of Ipecac, N.F., are
assayed for total ether-soluble alkaloids by procedures which
involve the sllme principles as the as flay of ipee1w.
Questions and Problems
1. Why is ipecac assayed for its eOlltcllt of ether-soluble' alkaloids?
2. Ascertain the IIlolecl!la1' weights of emetine arid ~cphae1iJle from '"he '
tflcble, U.S.P. page 587, and Hhow how the alkaloidal cqu~valen~ of c!Lch
cubic centimeter of 0.1 N H 2SQ,1 is derived. " ' "
3. Would the error in the lLssay result be very considerable "if the 'etl~er~
soluble ~lkaloids of the ipecac cOll1:listed almost ~ntirely of emet{~e? ".- ',.' ,
4. Enumerate several possible sources of e~ror which might be e~j(joullterad
in the assay of ipecac. .t """,' ;,

Exercise 118
Object.-Assay of Areca. _. ,
Materials Required.-15 Gm. of !Ll'eea.
250 cc. of ether.
15 of a saturated solution of sodium biclLl'bonate (!\bout 1 in 10).
About 3 Gm. of sodium bicarbonate. >
Procedure.-l. "Pla,ce 15 Gm. of Areca, in moderately fine powder and
accurately weighed, into a suitable flask, add 150 ce. of ether, allow to stand
about five minutes, then add 15 co. of a saturated solution of sodium bicar-
410 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

bonate. Agitate the mixture in a mechanical shaker for one hour, or inter-
mittently during two hours, set it aside overnight, and again agitate in a
mechanical shaker for one hour. Separate 100 cc. of the clear ethercal
liquid." (See General Procedure 3A, page 395.)
The most important alkaloid occurring in areca is arecoline.
A number of other alkaloids, namely, arecaidine, ethyl arecaidine,
guvacine, iso-guvacine, guvacoline, and arecolidine, have also
been found to b~ present in the drug in small amounts. The
alkaloids which are largely combined as salts in the drug are
liberated by the sodium bicarbonate. The arecoline and
guvacoline dissolve in the organic solvent while a large proportion
of the other alkaloids, e.g., guvacine and arecaidine, remain in
the aqueous alkaline liquid because they contain carboxy groups
and form water-soluble salts with alkalies. Since arecoline is
the methyl ester of arecaidine and guvacoline is the methyl ester
of guvacine, the carboxy groups are blocked, and these alkaloids
do not form salts with alkalies.
2. "Extract the alkaloid completely from the ethereal solution with suc-
cessive portions of dilute ~ulfuric acid (about 1 per cent). Render the
combined acid solutions IIlkaline with sodium bicarbonate, cautiously
added, to avoid loss by effervescence." (See General Procedure 4, page
398.)

The arecoline and guvacoline which are soluble in all propor-

tions in water and in ether form salts of the acid which are soluble
in water but very slightly soluble in ether.
3. II Extract the alkaloid completely from the alkaline liquid with succes-
sive portions of ether. Filter the ethereal solutions successively into 10 cc.
of tenth-normal sulfuric acid, rotating after each addition. Evaporate the
ether from this mixture, and determine the alkaloid by titration of the
excess acid with fiftieth-normal sodium hydroxide, using cochineal T.S. or
methyl red T.S. as t,he indicator.
"Each cubic centimeter of tenth-normal sulfuric acid represents 0.0155
Gm. of arecoline, CsH 1a02N." (See General Procedures 5, 6, and 7
page 399.)

The free alkaloids dissolve in the ether. When the ethereal

solution is added to the acid and evaporated, the alkaloids form
salts with a part .of the acid and the amount of acid in excess is
determined by titration.
2C sH 130 2N + H2S04-(C8H1302N)2~H2S04
 OFFICIAL TYPE METHODS 411

The National Formulary requires that areca contain not less

than 0.2 per cent of alkaloids calculated as arecoline. Calculate
the pel' cent of alkaloids contained in the sample assayed.
TABLE LXI.-DnuGs AND PUEPARATIONS ASSAYED BY THE ALIQUOT-PART
METHOD .

Amount
Official
used, Assayed
Substance requirement,
Gm. for
per cent
or cc.

U.S.P.
Cinchona ............... 5 Total alkaloids (n.l.t.) 5
Cinchona, compound tinc-
ture of ....0 , " 50 Total alkaloids 0.4toO.5W/V
Ipecac ................. 10 Ether-soluble (n.1.t.) 2
alkaloids
Ipecac, fluidextract of.. 0 0 10 Ether-soluble 1.8to2.2W/V
alkaloids
N.F.
Areca .......... 0 0 15 Arecoline (n.Lt.) O. 2
Hydrastis ............... 10 Ether-soluble (n.Lt.) 2.5
alkaloids
Hydrastis, extract of..... 2* Ether-soluble 9 to 11
alkaloids
Hydrastis, fluidextract of 5* Ether-soluble 2.25to 2.75W/V
alkaloids
Hydrastis, tincture of .... 25* Ether-soluble 0.45 to 0.55W/V
alkaloids
Ipecac, tincture of ....... 50* Ether-soluble 0.I8to 0.22W/V
alkaloids

(noLt.) = not less than.

* No aliquot pa.rt taken.
Alkaloidal Assays by the Total-extraction Method
Exercise 119
Object.-Assay of Hyoscyamus for Total Alkaloids.
Materials Required.-25 Gm. of hyoscyamus in No. 60 powder.
10 cc. of alcohol.
250 cc. of ether.
150 cc. of chloroform.
Procedure.-l. "Place 25 Gm. of Hyoscyamus, in fine powder, in an
extraction thimble, insert the thimble in a Soxhlet extractor, moisten the
412 QUANTI'l'Jt'j'IVE PHARMA.CEUTICAL CHEMISTRY

drug with a mixture of 8 ce. of stronger ammollill T.S., lOce. of a1t:ohol and
20 ce. of ether, and mix thoroughly. Macerate the mixture over night, then
extract it for not less than three hours on !t water bath, using ether as the
solvent. The following alternative pro(:ess lllay be used: Moisten 25 Gm.
of HyoRcyalllu~, in fine powder, with fL mixture of 8 ec. of stronger ammonia
T.S., 20 e~. of ethel' and 10 ce. of ehloroform, in a small pereolator, previously
prepared by packing the outlet with a pledget of purified cotton. Macerate
the mixture over night, pllek it iu the percolator and extmct by slowly per-
colating with a mixture of.-3 parts of ethel' und 1 part of chloroform, by
volume. Continue the percolation until the 3 or 4 ce. of percolate last
passed, when eV!1pomted to dryness and the residue dissolved in dilute
sulfurie !tcid, fails to beeome turbid when treated with mercuric potassium
iodide 'f.S." (See General Procedure 3B and C, page 397.)

Hyoscyamine, the chief allG1loid of hyoscyamus, and small

quantities or" other alkaloids are set free from the acids, with
which they are combined ih the crude drug by the ammonia, and
dissolve in the organic solven,t. The laevorotatory hyoscyamine
is largely converted into its racemic isomer, atropil1:e, in the
ammoniacal solution.
2. "Evltpol'[l,te the extractive, obtained by oithcr mothod, to about 15 co.,
then add 10 ec. of approxirnntcly tenth-normal sulfuric acid and 10 ec. of
distilled wn.ter, und continue the evaporation until tho volatile solvents are
removed. li'iU(lr this mixture, collecting the filtrate in [1 separator, disRolve
the chlorophyll residue in chloroform, add acidulated water, evaJJorate on 11
water bath until the chloroform is removed, ::tnd filter into the same separator
through the filter previouRly uRcd."

The large volume of liquid obtained, especially by cold percola-

tion, is concentrated to a small volume to se(Jure a convenient
amount. It should not be evaporated to dryness because the
chlorophyll and other non-alkaloidal residue make it difficult
to completely diss~lve the alkaloids in the acid. The alkaloids,
hyoscam-ine and atropine, being esters undergo hydrolysis readily
in aqueous acid and alkaline solution especially when heated.
Consequently, the evaporation of the organic solvent shmlld be
. "
conducted at a low temperature, preferably not exceeding 40C.,
after the addition of the acid. The evaporation may be Carried
out advantageously in a vacuum desiccator under reduced pres-
sure. Upon filtration, the non-alkaloidal residue consisting
mostly of chlprophyll is retained on .the filter. This residue is
diss~ly'ed in chloroform, treated with acid, and evaporated to
 OF[flCIAL TYPE METHODS 413

completely extract the alkaloids. This method of obtaining

the alkaloids in acid solution from the orgn,nic soh'cnt avoids
the formation of the stable emulsions frequently obtained by the
shaking out process. Quite stable emulsions are particularly
liable to be encountered in shaking out alkaloids in the assay of
hyoscyamus, belladonna, and stramonium owing to the presence
of small amounts of saponins which act as emulsifying agents.
The sulfates of the alkaloids are formed and dissolved in the
acid solution leaving most of the coloring matter, etc., in the acid
insoluble residue
3. "Render the mixed filtrates alkaline with ammonia T.S. and remove
the alkaloids by extracting with chloroform, tc~ting for the complete extrac-
tion of the alkaloids." (See General Procedure 5, page 399.)

The alkaloids are set free by the ammonia water and dissolve
in the chloroform.
4. "Evaporate or distil the chloroform from the combined extractions
until reduced to a small volume, then evapomte to dryness on a water bath,
and keep at this temperature for fifteen minutes. Dissolve the residue in
chloroform, evaporate to dryness on It water bath, and eontinue the heating
for fifteen minutes. Repeat this treatment for the third time."
The solution of the alkaloids in organic solvent is evaporated
to drive off the solvent and ammonia. The residue is repeatedly
dissolved, evaporated, and dried to drive off small amounts of
volatile amincs which are basic and, unless removed, would
neutralize some of the acid in the next step of the procedure with
subsequent high results.
5. "Dissolve the resulting residue in chloroform, add 15 ce. of fiftieth-
normal sulfuric acid, remove the chloroform by evaporation, and titrate the
excess acid with fiftieth-normal sodium hydroxide, using methyl red as the
indicator. Each cubic centimeter of fiftieth-normal acid is equivalent to
Q.00578 Gm. of the alkaloids of Hyoscyamus."
The residual alkaloids, which consist chiefly of the isomers
atropine and hyoscyamine, react with some of the sulfuric
acid as follows:
2C17H2S0sN + H2S04---?(C17H~sOaNh.H2S04
289.19 98
Since 98 Gm. of H 2S0 4 is equivalent to 2 X 289.19 Gm. of
atropine or hyoscyamine, each cubic centimeter of 0.02 N H 2S04
414 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

is equivalent to 2 ~ ~02~9;~00 = 0.00578 Gm. of the alkaloids

of hyoscyamus. If exactly 10 Gm. of the drug is used as the
sample for assay, the percentage of alkaloids in the hyoscyamus
cc. 0.02 NH 2S0 4 X 0.00578
. may be calculated as follows: 10 X 100.
or cc. 0.02 N H 2S0 4 X 0.0578 = percentage of alkaloids in the
sample.
The U.S.P. requires that hyoscyamus leaves contain not less
than 0.3 per cent of aikaloids. Calculate the percentage of
alkaloids contained in the sample assayed and compare the
results with the official requirements.
Belladonna leaf, belladonna root, and stramonium are assayed
by the same method as hyoscyamus with the exception that
smaller samples are used because these drugs contain larger
amounts of alkaloids.
Assay of Preparations of Hyoscyamus, Belladonna, and
Stramonium
Tinctures.-The tincturcs of hyoscyamus, belladonna (leaf),
and stramonium of the U.S. Pharmacopoeia are all assayed by
the same method except that the amounts of the tinctures
employed as samples are designated so that approximately
0.030 Gm. of the alkaloids will be obtained in the assay residue.
The assay method is the same as that for hyoscyamus except
that the extraction of the alkaloids is omitted since they are
already in solution. The assay process is illustrated in the
following exercise.
Exercise 120
Object.-Assay of Tincture of Belladonna.
Materials Required.-100 cc. of tincture of belladonna.
250 cc. of ether.
150 cc. of chloroform.
Procedure.--:-Aceurately measure 100 cc. of tincture of belladonna and
evaporate it, at a temperature not exceeding SOC., to a volume of about
10 cc. Add 10 ce. of approximately 0.1 N sulfuric acid and 10 cc. of dis-
tilled water. Filter this mixture, collect.ing the filtrate in a separator, dis-
solve the chlorophyll residue in chloroform, add acidulated water, evaporate
on a water bath until the chloroform is removed, and filter into the same
separator through the filter previously used. Proceed as in Step 3 of the
Assay of Hyoscyamus (page 413).
 OFFICIAL TYPE METHODS 415
Pilular and Powdered Extracts.-Thc pilular and powdered
extracts of hyoscyamus, belladonna, and stramonium are assayed
by the same method with the exception that different amounts
of the extracts are used. The following procedures illustrate
the methods: assay of pilular extract of hyoscyamus.
"Dissolve approximately 5 Gill. of Pilular Extract of Hyoscyamus, accu-
rately weighed (see Proximate Assays, page 39.5, for method of weighing),
in 10 cc. of chloroform, add 10 ee. of approximately tenth-norm!ll sulfuric
acid and 40 ec. of distilled water, and evaporate the mixture on a water hath
until the chloroform is removed. Complete the assay as directed under
Hyoscyamus, page 412, beginning with the words 'Filter this mixture,
collecting the filtrate in a separator.' Each cubic centimeter of fiftieth-
normal acid is equivalent to 0.00578 Gm. of the alkaloids of hyoscyamus."

Assay of powdered extract of hyoscyamus.

"Digest I1pproximately 5 GIll. of Powdered Extract of Hyoscyamus,
accurately weighed, in 10 ce. of chloroform, add 10 ce. of approximately
tenth-normal sulfuric acid and 40 ce. of distilled water. Mix well, filter
through purified cotton wetted with weak sulfuric acid, wash the cotton and
the residue with acidulated distilled water until free from alkaloids, discl1rd
the cotton and the residuc, and evaporatc the filtrate on a water bath
until the chloroform is removed. Complete the assay as directed under
Hyoscyamus, page 412, beginning with the words 'Filter this mixture,
oollecting the filtrate in a separator.' Each cubic centimeter of fiftieth-
normal sulfuric acid is equivalent to 0.00578 Gm. of the alkaloids of
hyoscyamus."

Fluidextracts.-Fluidextract of belladonna root of the U.S.

Pharmacopoeia is assayed by a method similar to that for
hyoscyamus as indicated by the following procedure:
"To lOcc. of Fluidextract of BeJladollna Root add 10 cc. of approximately
tenth-normal sulfuric acid and 10 cc. of distilled water, and evaporate on a
water bath to about 10 ee. Then add 10 ce. of distilled water and proceed
as directed under the Assay of Hyoscyamus, page 412, beginning with the
words 'Filter this mixture, collecting the filtrate in a separator.' Each cubic
centimeter of fiftieth-normal acid is equivalent to 0.00578 Gm. of the alka-
loids of belladonna."
Fluidextracts of belladonna leaf, hyoscyamus, and stramonium
of the National Formulary are all assayed by the same method.
The method described in the National Formulary differs some-
what from the method of the Pharmacopoeia as is shown in the
following exercise.
416 QUAN1'ITA'l'lVN J>J1AllMAC'EU1'lCAL CHEMIS'l'RY

Exercise 121

Object.-Assay of Fluidextract of Belladonna Leaf.

Materials Required.-l0 ceo of fluidextract of belhLdonno.leaf.
150 ce. of chloroforlll.
Procedure.-"To 20 ce. of chloroform in [\ sepamt.ory funnel, add 10 ce.
of Fluidextract of Belladonna Leaf, accurately measured, 10 ce. of water, !Illd
:1bout 1 ce. of ammonia water, and shake the mixture vigorously for one
minute. Separate the chloroformic layer, and complete the extraction of
the alkaloids with successive portions of chloroform. Extract the alkaloids
completely from the combined chloroformic solutions by shaking with suc-
cessive small portions of dilute sulfuric acid (about 1 per cent). Filter the
acid solutions successively through a small filter or pledget of cotton into a.
sepamtory fUIlllel. Add a slight excess of ammonia water, and completely
extract the alkaloids from the aqueous bye I' by shaking with successive

TABLE LXII.-AJ,I(ALOIDAL AHi:lAYS BY 'l'lIE TOTAL EXTRACTION PROCESS

Amount
Official
used,
Su)mtance requirement,
Gm.or
pcr eent
ce.

U.S.P.
Belladonna leaf ................... . 10 (n.l.t.)0.3
Belladolllla ointment .............. . 25 0.118 to O. 132
Belladonlla, pilular extract of ...... . 3 1.18 to 1.32
Belladonna plaster ................ . 10 0.25 to 0.30
Belladonna, powdered extract, of ... . 2 1.18 to 1.32
Belladonna root ................... . 10 (n.Lt.) 0.45
Belladonna root, fluidextrac~ of .... . 10 0.405 to 0.495W/V
Belladonna, tincture of ............ . 100 0.027 to 0.033W/V
Hyoscyamus ..................... . 25 (n.l.t.) 0.040
Hyoscyamus, pilular extract of ..... . 5 0.135 to 0.175
Hyoscyamus, powdered extract of .. . 5 0.135 to O. 175
HY9scyamus, tincture of ........... . 250 0.0034 to 0 .0046W /V
Stramonium ...................... . 10 (n.Lt..) 0.30
Stramonium, pilular extract of ..... . 3 1.10 to 1.30
Stramonium, powdered extract of .. . 2 1.10 to 1.30
Stramonium, tincture of ........... . 100 0.027 to 0.033W/V
N.F.
Belladonna leaf, fluidextract of ..... . 10 0.27 to 0.33 W/V
Hyoscyamus, ftuidextract of ....... . 25 0.035 to 0.045W/V
Stramonium, fluidextract of ........ . 10 0.25 to 0.35W/V

(n.l.t.) = not less than.

 OFFICIAL TYPE METHODS 417
portionR of chloroform. Evaporate the combined chiorofoflll\<;> ,;olutinl\s to
drynE'~H and hellt the dry residue on [1 wn.trr bath for 15111inut~s. TI1PU add
about 1 ce. of neutral aleohol, again evaporate to uryne~s, and lICat thp resi-
due on t1 water hath for Iii minutes. Mix the residue with 2 cr. of neutral
alcohol, adrl 15 ce. of fiftieth-normal sulfuric acid, and warm the mixture to
dissolve the alkaloids. Cool, ancj. titrate the ('xcess acid with fiftieth-normal
sodium hydroXIde, using methyl rpa T.S. or cochineal T.S. as the indicn.tor.
"Each cubic cC'utimeter of fIftieth-normal sulfuric acid is equivalent to
0.005784 Gm. of the alkaloid~ of BelladoIln!1 Leaf."

Ointment of Belladonna and Belladonna Plaster.-Ointment

of belladonna and belladollnfi, plai:lter of the U.S. Pharmaropoein.
are assayed by special methods designed to tleparatc the alkaloids
from the ointment 01' plaster base. The principles involved are
similar to those already consido~ecl.
Questions and Problems
1. What isomcrit 'change does the chief alkaloid 1;I bE'lladonna undergo
in the assay process?
2. ThE' alkaloids of belladonna have the structure of esters. Wllat type
of decomposition might thry undE'rgo if allowed to stand in eontact for !1
long time with a strong alkali?
3. How much 0.1015 N II,SO. will be required to form lhe sulfates of
the alkaloids from 25 Gm. of hyoscyamus leaves which contain 0.070 per
cent alkaloids?
 CHAPTER XXVI

ALKALOIDAL ASSAYS BY SPECIAL METHODS

A number of the official alkaloidal drugs and their preparations
cannot be assayed by any of the type methods previously dis-
cussed because of the nature of the drug or of the properties of
the alkaloids to be determined. Drugs of this type are assayed
by special methods.
The assays of opium and its preparations differ from those of
the other alkaloidal drugs, due to the fact that morphine exhibits
exceptional solubility properties; i.e., it is practically insoluble
in most organic solvents.
Exercise 122
Object.-Assay of Opium.
Materials Required.-6 Gm. of opium.
3 Gm. of freshly slaked lime.
1 Gm. of ammonium chloride.
100 cc. of ether.
25 cc. of water saturated with morphine.
Procedure.-l. "Triturate in a mortar 6 Gm. of the Opium to be assayed
which, if fresh, should be in very small pieces, and if dry, in fine powder, with
about 40 cc. of distilled water for fifteen minutes. Transfer the mixture
completely to a :Bask with the aid of 30 cc. of warm distilled water, stopper
the flask, and shake it every ten minutes, or continuously in a mechanical
agitator, dUTing one hour. Pour the contents as f'venly as possible upon a
wetted filter of from 10 to 11 cm. in diameter or, preferably, upon a sintered
glass funnel attached to a suction flask. When the liquid has drained off,
wash the residue with about 20 cc. of distilled water, carefully dropped upon
the edges of the filter and its contents. Transfer the moist residue to a
mortar, rub it to a smooth paste, then rinse it into the original flask with
40 cc. of distilled water, agitate it thoroughly during ten minutes, and return
the entire mixture to the filter. When the liquid has drained off, wash the
residue with small portions of distilled water until the washings are nearly
colorless."

The alkaloids of opium, about 25 of which are known, occur in

the drug as meconates and sulfates. When the opium is agitated
41R
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 419

and macerated with distilled water, most of the meconates and

sulfates of the alkaloids are dissolved. Complete extraction
of the alkaloids is obtained when the residual opium is treated
with a second portion of distilled water. By rubbing to a smooth
paste, any lumps are broken up so that the water can come in
contact with all of the particles of the drug. The residue is
washed with distilled water after each extraction to displace all
of the solvent absorbed by the opium.
2. "Evaporate the fIltrates and w1tshings, in a t1tred dish, to 1tbout 30 Gm.
and allow to cool. Now add 3 Gm. of freshly slaked lime, triturate the
mixture for fifteen minutes, and transfer it completely to a tared flask with
the aid of small portions of distilled water. Then add sufficient distilled
water to make the mixture weigh 54 Gm. and mix welL Filter the mixture
using a filter from 10 to 11 em. in diameter, which has not been previously
moistened, collecting the filtrate in a dry cylinder or small flask. During
the filtration keep the funnel covered with a glass plate and its stem well
within the neck of the receiving vessel."
The combined aqueous filtrates are evaporated on a water
bath to a volume of about 30 cc., since the alkaloids which are
quite stable at 100 e. might be decomposed if the evaporation
0

was carried out over a direct flame. The purpose of concentrat-

ing the aqueous alkaloidal extractive is to reduce the volume to
a convenient working quantit.y. The concentrated extract is
made up to a weight of exactly 54 Gm., so that an aliquot portion
representing a definite weight of opium may be taken. When
the aqueous opium extract is t.riturated with freshly slaked
lime (prepared by adding dropwise 1 cc. of water to 3 Gm. of
quicklime, allowing the product to become pulverulent, and cool),
the alkaloids are liberat.ed from their salts and the morphine,
codeine, and narcotine are dissolved in the alkaline calcium
hydroxide solution, most of the other alkaloids of opium remaip-
ing as insoluble matter along with t.he calcium meconate, calcium
sulfate, resins, etc. The mixture may. be transferred to a flask
and shaken for 15 min. to allow the reactions in which the alka-
loids are liberated to go to completion, the flask being stoppered
to prevent evaporation so that the volume of liquid will not
change appreciably, since an aliquot portion of it is employed in
the assay.
3. "Place 34 Gm. of this filtrate (corresponding to 4 Gm. Of Opium) in an
Erlenmeyer flask of suitable capacity, add 2 ce. of alcohol and 15 cc. of ether,
420 QUAN'l'ITATIVE PHARMACEUTICAL .CllEMIS'1'RY

and, after shaking the mixture, add 1 Gill. of ammonium chloride. Stopper
the flask, shake it frequently during.ten minutes and set it aside in a cool
place over night. Remove the stopper and brush any adhering crystlLls back
into the flask."

Since the total aqueous extract containing the alkaloids

weighed 54 Gm. after adding the lime, 54 - 3 = the weight of
liquid and an aliquot portion of tWo-thirds of this liquid or 34 Gm.
eontains an amount of the alkaloids corresponding to % of 6 ~
4: Gm. of opium.
Ammonia is liberated from ammonium chloride:

2NHDl + Ca(OHh_"2NH~OH + CaCh

and morphine is precipitated, since it is insoluble in ammoniacal
solution and in ether. The alcohol causes the morphine to
separate out in the form of relatively large crystals; whereas, .
when alcohol is not used the alkaloid comes down in the form of a
fine powder. The former are more easy to colleet and less rapidly
soluble in ether than the powder. The mixture is allowed to
stand in a cool place for 12 hr. to allow the larger crystals to grow
at the expense of the smaller ones. The codeine and nareotine
remain in solution in the ether.
4. "Dccl1nt the ethereal layer through a small filter paper. Rinse the
flask !1nd contents with 15 cc. of ether and pass these washings through
the filter, and again wash the filter with an additional small quantity of
ether. When all of the ether has p!1ssed through the filter, pour the aqueous
layer upon the filter. without trying to remove Lhe crystllis from the fla~k.
Wash the crystals in the flask !1nd the contents of the filter with distilled
water, previously satunl,ted with morphine, unW the w!1shings are colorless.
Then add a few drops of distilled water to displace the morphinated water."

The crystals are washed with ether to remove any traces of

eodeine or narcotine, since these alkaloids are insoluble in ammo-
nia water but soluble in ether. Water saturated with morphine
is used to wash th~ crystals to prevent solution of the crystalline
morphine which is very slightly soluble in water. This aqueous
solution removes any ammonia present as well as any other
water-soluble impurity adhering to the surface of the orystals,
such as gums, 9010ring matter, etc. The morphinated water
must be displaced from the paper filter with distilled water
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 421

to avoid including in the final result some of the morphine

from the morphinated water.
5. "Add to the flask containing the crystals about. 10 cc. of hot neutral
methanol, agitate to dissolve 118 much of the morphine as possible, :md pour
the solution over the morphine en the filter, receiving the filtrate in a suit-
able flask. Repeat this treatment with hot, neutral methanol three or four
times, using from 5 to 7 cc. each time, until all of the morphine has been dis-
solved. Cool this solution and add exactly 25 cc. of tenth-llormal sulfuric
acid. Dilute this solution with 50 cc. of distilled water and titrate the
excess of acid with tenth-normal sodium hydroxide, using methyl red T.t!.
as the indicator, Each cubic centimeter of tenth-nomllLl acid is equivulent
to 0.02852 Gm. of anhydrous morphine."
..
The morphine crystals dissolve in the 0.1 N sulfuric acid with
the formation of morphine sulfate:
2C 17H 190 3N + H2S04----7(C17H190aN)dlzSO'I.
2 X 285.16 98
One thousand cubic centimeters of N sulfuric acid is equiva-
lent therefore to 285.16 Gm. of anhydrous morphine, and each
cubic centimeter of 0.1 N sulfuric acid is equivalent to
10 2!5i~~00 = 0.02852 Om. anhydrous morphine.
The Pharmacopoeia requires that opiulll contain not less thall
9.5 per cent of anhydrous morphine. Calculate the percentage
of anhydrous morphine found in the sample of opiulll assayed
and compare the resliits with the official requirements.
Granulated opium and powdered opium are assayed in exactly
the same manner as described above. The general procedure
used in the assay of tincture of opium is also the same, except
that 60 ee. of the tincture is evaporated to remove the alcohol,
diluted to exactly 54 Gm. with distilled water, and treated with
lime, etc., as dirceted in steps 2 to 5 of the above assay procedure.
Camphomt!3d tincture of opium is assayed by a special method.
Extract of opium is also assayed by this method, but in the first
step of the procedure, a 3 Gm. sample of the extract is dissolved
in water and the solution is diluted to 54 Gm. after the addition
of tlw lime, etc.
Questions and Problems
1. What functions do the calciuIll. oxide and I1Xlllllonium chlQride serve
in the above assay?
422 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

2. One gmm of morphine is soluble in about 7,632 cc. of ether at. 250.
I,ook up the solubilit.y of codeine in the U.S.P. and explain how ether may
be used to separfLte the nlkaloids.
3. Why is water saturated with morphine used to wash the crystallized
morphine?
4. Morphine contains a phenolic hydroxyl group. Explain why it is
appreciably soluble in alkaline solutions.

TABLE LXII I.-OFFICIAl, SUBSTANCES ASSAYED BY THE OrIUM ASSAY

METHOD

Amount. Anhydrous mor-

Equivalent
used, phine, ollicial
Substance of 1 cc. of
GIll. or requirement,
0.1 N H 2SO 4
cc. per cent

U.S.P.
Opium .................. 6 0.02852 9.5
Opium, granulated ........ 6 0.02852 10 to 10.5
OpiuIIl, llowdcred ......... 6 0.02852 10 to 10.5
Opium, tincture of ........ 60 0.02852 0.95 to 1.05WjV
Opium, tincture of, C~lIn-
phorated .............. 100 0.00570* 0.035 to 0.0045
N.F.
Opium, extmct ........... 4 0.02852 19.5 to 20.5
- * Equivalent of 1 CC. of 0.02 N H,SO .

Exercise 123

Object.-Assay of Colchicum Seed.

Materials Requii:ed.-15 Gm. of colchicum seed in No. 60 powder.
10 cc. of solution of lead subacetate.
2 Gm. of sodium phosphate.
150 cc. of chloroform.
20 cc. of ether.
Procedure.-l. "Place 15 Gm. of Colchicum Sceel, in fine powder, in It
suitable flask and add 290 cc. of distilled water and 10 cc. of lead subacetate
1'.8. Weigh the flask and contents, and digest the mixture at from 60 to
70C. for three hours with occasional agitation. Cool, add distilled water
to restore the original weight, and filter, collecting 200 cc. of filtrate."

Colchicine is the only medicinally important constituent of

colchicum. Both the seed and corm, which are assayed by
the same method, contain a considerable amount of starch,
sugar, gum, tannin, resin, fatty oil, and coloring matter from
 ALKALOID.flL ASSAYS BY SPECIAL METHODS 423

which the alkaloid must be s('parated. The free alkaloid, col-

chicine, is miscible in all proportions with water, and it is extrnctrd
from the drug \vhen the latter is digested with water. The
digestion temperature should not exceed 70C., heeam,e the
starch content of the drug, espee:ially of the corm, is gehtinized
at higher temperatures, and because colchieine if; IPss soluble in
hot than in cold water. The lead subneetate is added to prevent
solution of the tannin, gums, resin, and eolOl:illg matter with
which it forms water-insoluble compounds and to liberate the
colchicine from acids with which it may be combined in the crude
drug. The mixture is cooled, and distilled water is added to
restore the originttl weight uf the Rample, waterJ and lead :-lub-
acetate mixture, so that all aliquot portion, eorrespouding tu a
definite weight of the sample taken, may be used .. Two hundred
cubic centimeters of the liquid representing the alkaloidfl con-
tained in two-thirds of the weight of the sample are filtpred off.
2. "Add 2 Gm. of sodium pho8phrtte to the clear filtrat.e, or sufficient to
precipitate the lead completely, shake the mixture frequently during hnlf an
hour, filter, collect 100 ce. of filtrate, representing 5 Gm. of Colehicum Seed."

The sodium phosphate precipitates the excess lead Sllhaeetate

from the solution as insoluble lead phosphate, leaving sodium
acetate in solution. A second aliquot portion of 100 ceo corre-
sponding to one-third of the total sample used or 5 Gm. of col-
chicum is taken.
3. "Complet.ely ext.ract. the alkaloid from the filtrate with chloroform, as
shown by testing with iodine T.S., and evapor!lte the solution in It tared
flask. Add 1 cc. of alcohol, and again evaporate. Repeat ihis operation
once more, and dry the residue to constant weight. at 1000."

Colchicine is miscible ill all proportions with chloroform, and

this solvent extracts the alkaloid, which is very feebly basit',
from neutral or acid solutions. The alkaluid forms an addition
product of unknown cOIlstitution of the formula C 22 H250oN.-
2CHCl a with chloroform. The chloroform is driven off froIn this
addition product when it is treated with successive portions of
alcohol and evaporated to dryness at 100C. The residue, dried
to constant weight at 100C. and weighed accurately, consists
chiefly of crude colchicine and colchiceine ..
424 QUANTITATIYE PHARMACEUTICAL CHEMISTRY

4. "To this weighed residue in the flask, I.1dd 5 cc. of tenth-normal sulfuric
!Icid, 5 cc. of distilled water, and a few drops of chloroform, nud hent the
mixture at 70C. for ten minutes. Filter the liquid through a pledget of
purified cotton, wash the flask and cotton with distilled water, reject the
filtrate and washings, and remove as much of the water from the cotton lIS
possible. Dissolve the residue, if any, which may remain on the cotton by
washing it first with II little alcohol and then with ethel'; collect the alcohol-
ether washings ill the flnsk, evaporate, and dry the residue to constant.
weight. at lOO~C. De(luct this weight from the weight of residue previously
obtained. The difference is the weight of colchicine obtained from 5 Gm.
of Colchicum Seed.'"

A dilute solution of sulfuric acid heated to 70C. is used to

dissolve the colchicine completely from the residue, since this
alkaloid decomposes rapidly when heated to 100e, with strong
mineral acids. The addition of a few drops of chloroform
facilitates the solution of the colchicine from the residue. The
colchicine in acid solution is filtered off through cotton and
removed from the filtration medium by washing with distilled
water, leaving acid-insoluble impmities on the cottOll and in the
flask. The impurities are dissolved by successive treatment
with alcohol which is miscible with water and with ether. The
total residual impurities, when dried to constant weight at the
same tempemture as the crude colchicine and deducted from
the 'weight of the latt.er previously obtained, gives the weight of
colchicine which dissolved in the acid solution; e.g., if an aliquot
portion corresponding to 5 Gm. of colchicum yields 0.025 Gm. of
crude colchicine and the residual impurity is found to weigh
0.025 - 0.005 ..
0.005 GIll., then 5 X 100 = 0.4 per cent eolchlCme
contained in the sample.
Colchicum seed must contain 0.45 per cent of colchicine to
meet the official requirement. C!1lculate the percentage of
colchicine contained in the sample assayed and compare the
result with the official requirement.
Colchicum corm is assayed in the same way as colchicum seed.
The extract, fluidextract, and tincture are also assayed by the
same method, but in the case of the fluidextract, 15 cc. is taken
as the Rample and 275 cc. of water is used to bring the volume of
the liquid to 290 ce., so that when 10 cc. of lead subacetate is
added, the total volume of the mixture will be 300 cc. In the
 ALKALOIDAL ASSAYS BY SPECIAL METHODS' 425

assay of the tincture, a volume of 150 ec. is taken, c\Taporated to

15 cc., and made up to 300 cc. by the t1ddition of 275 cc. of dis-
tilled water and 10 cc. of lead subacetatc solution.
Questions and Problems
1. 'What exceptional solubility properties are exhibited by colchidne'?
2. Why is lead subacetate used in the extraction of the 1111mloid from
the crude drug?
3. Why should not the aqueous mixture be heated ahove 70C. in the
extraction of colchicine from the powdered dr11g?
4. What property of colchicine makes it impossible to determine this
alkaloid accurately by volumetric methods'?
5. Why should the crudc colchicine not, be heated above 70C. with the
0.1 N sulfuric acid?

TABLE LXIV.-OFFICIAL SUBSTANCES ASSAYED BY THE SAM:E METHOD AS

COLCHICUM:

Aliquot por- ColnhicillP., official

Amount
Substance tion corre- requirement"
useel
spol1ds to per cent
-----
U.S.P.
Colehicum seed ............ 15 Grn. .') Gm. 0.45
Colchiwm seecI, tinct.ure of 150 ec. 50 ee. 1.036 to 0. 044lV /V
N,F.
Colchicum corm. , ......... 15 Gm. S Gill. O.3S
Colchicum corm, fluidex-
tract of ................. 15 ce. 5 ce. 0.31 to O.39lV/V
Colchicum corm, strong
tincture of .............. 45 ceo
Colchicum seed, fluidextmct
15 eo.
.0.125 to 0.155WjV
of .............. , .. " .. 15 cC. 5 ce. 0.4 to O. .5W/V

Exercise 124

Object.-Assay of Nux Vomica.

Materials Required.~l0 Gm. of nux vomica in No. 60 powder.
100 cc. of ether-chloroform mixture.
100 cC. of chloroform.
Procedure.-l. "Place 15 Gm. of Nux Vomica, in coarse powder, in a
flask 01' bottle, add 150 cc. of a mixture of 3 volumes of ether and 1 volume
of chloroform, agitate the mixture, and allow it to stand for about two
minut.es. Then add 10 cc. of stronger ammonia T.S., agitate thoroughly,
stopper the container securely and shake frequently, but gently, during one
426 QUANTITATIVE PH.r1RMACEUTICAL CHEMISTRY

hour. Now allow the mixture to stand for twelve hours or over night in a
cool place .. At the expiration of this period, shake the container gently for
fifteen minutes, and then allow the liquids' to separate."

The alkaloids of nux vomica which consist principally of

strychnine and brucine are set free by the ammonia water and
dissulve in the ether-chloroform mixture.
2. "Decant 100 ceo of the liquid (representing 10 Gm. of Nux Vomica),
and trnnsfer it, to a, ~eparator, rinsing the container with a little chloroform
and adding the rinsings to the separator. Now [Idd about 40 ceo of approxi-
mately normal sulfuric acid to the separator and shake the mixture gently
for five minutes, then allow the liquids to separate, and draw off the acid
layer into another separator and repeat with successive port.ions of the acid,
until the ethereal liquid is completely extracted." (Test for the complete
extraction of the alkDJoids as directed on pllge 399.)

The alkaloidal sulfates are formed and dissolve in the acid

solution; fats, resins, coloring matter, etc., remain in the ether-
chloroform layer.
3. "To the combined acid solutions in the separator, add a small piece of
red litmus paper and 50 cc. of chloroform, and follow with sufficient ammonia
1'.8. to render the aqueous layer alkaline and, aft.er gently shaking, add 2 or
3 cc. more of the ammonia 1'.8. Now slmke the mixture thoroughly, but
gently, for about; ten minutes, a.nd allow the liquids to separat.e. Draw off
the chloroform int.o a contniner and repellt, with additional portions of
chloroform, until all of Lhe alkaloid is extracted."

The alkaloids are liberated and dissolve in the chloroform.

4. "Carefully evaporate the combined uhloroforrnic extracts to dryness on
a water bath, dissolve the residue by warming with 15 ce. of approximately
3 per cent sulfuric acid, cool, and then add 3 ce. of a mixt.ure of equal parts
of nitric acid and a 5 per cellt solution of sodium nitrite in distilled water,
stir well, and allow to stand for exactly ten minutes at room temperature.
At the expiration of this period, pour the red solution into a separator con-
taining 50 co. of chloroform, rinse the flask with distilled water, and add the
rinsings to the separator. Now immediately add sufficient 10 per cent
sodium hydroxide solut,ion to t.he cont.ents of the separator to render it
distinctly alkaline to litmus paper, and then add a few cubic centimeters
more of the hydroxide solution."

The strychnine and brucine are converted to their sulfates by

the sulfuric acid. When the nitric acid and sodium nitrite mix-
ture is added, the brucine is oxidized to a non-alkaloidal, red
colored, quinonoid compound having phenolic properties. The
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 427

strychnine is very stable to oxidizing agents and is not attacked

in significant amounts during thc short period of oxidation. The
decomposition of the bruclne is completed in 10 min. If the
mixture were allowed to stand for a longer period, a small amount
of the strychnine would be oxidized. Consequently, the mixture
is neutra,lized with sodium hydroxide to stop the oxidation and
an excess of the alkali is addcd to liberate the strychnine. Some
workers prefer to liberate the strychnine by the addition of alkali
prior to the addition of the organic solvent because the complete
decomposition of the strychnine salts is difficult to accomplish
in the presence of organic solvents.
5. "Shake the mixture gently for ten minutes and allow the liquids to
separate. Draw off the chloroformic layer into another sepl1rator tutd
repeat the slJaking out with additional portions of chloroform until the
alkaloid is completely extracted. Add 10 ce. of distilled water to the com-
bined chloroformic extract, shake the mixture gently, and add a slllall piece
of red litmus paper. The litmus paper should indicr1te not more than a slight
alkalinity. If the water, after shaking with the chloroform, is strongly alka-
line, draw off the chloroform into another separator, and shako it with another
10 cc. of distilled water. Draw off the chloroform, passing it through a filter
paper moistened with chloroform, into a container. Wash the filter paper
with warm chloroform aud add it also to the container. Now shake out the
combined water extract with 5 cc. of chloroform and draw off all of the
chloroform through a chloroform-moistened filter paper as hafore."

The mixture is shaken for 10 min. to completely decompose the

strychnine salts. The liberated strychnine dissolves in the"
chloroform while the phenolic quinonoid oxidation product from
the brucine remains in the aqueous alkaline layer. Sodium
hydroxide rather than ammonium hydroxide is used as the alkali
because ammonium hydroxide is not a sufficiently strong base to
combine completely with the phenolic oxidation products and
prevent their passing into the chloroform. The combined
chloroform extractive is washed with water to remove any alkali
or alkali derivative of the phenolic oxidation prorlucts. This
aqueous wash liquid is then washed with chloroform to recover
any trace of strychnine.
6. "Evaporate the combined chloroform very carefully 011 a water bath
nearly but not quite, to dryness. Add to the moist residue 6 cc. of tenth-
normal sulfuric acid and follow by 30 cc. of distilled water. Heat the mixture
on a water bath until the alkaloid is dissolved and the odor of chloroform is
428 QUANTITATIYE PHAnMACEUTICAL CHEMISTRY

dissipated. Cool to room temperature and titrate the excess of acid with
fiftioth-normal sodium hydroxide, usiug 1 drop of methyl red T.S. as the
indicator. Each cubic centimeter of tCllth-normltl sulfuric acid is equivalent
to 0.03342 Om. of strychnine."

The chloroform should not be evaporated to dryness before

adding the acid since there is likely to be some loss of strychnine
by decrepitation if this is done. The strychnine is converted into
its sulfate quantitatively by the 0.1 N H2S04; the excess acid is
then determined by titration with 0.020 N NaOH. The equation
for the reaction is:
2CZ1H2202Nz + HzSOC--(CZ1H2202N2)2.H2S04
334.19

The number of cubic centimeters of 0.1 N H 2S0 4 found upon

residual titration with 0.020 N NaOH subtr:1cted from the
number of cubic centimeters of standard acid solution originally
added gives the quantity of standard acid solution which com-
bined with the alkaloids.
The U.S.P. requires that nux vomica con Lain not less than
2.5 pel' cent of alkaloids. Calculate the percentage of alkaloids
contained in the sample assayed and compare the result with the
official requirement.
TABL]i] LXV.-ASSAYS OF Nux VOMICA AND Nux VOMICA PREPARATIONS

Amount
used, Official reqlliremen t,
Substance
Gm. or per cent
cc.

U.S.P.
Nux vomica ... ,., ..... , ... ,.,. 15 St,rychnine = (!l.U.) 1.15
Nux vomica, extract.of ........ . 1.5 Strychnine = 7 to 7.75
Nux vomica, tincture of ....... . 100 Strychnine = 0.1.08 to 0.120
N.F.
Nux vomica, iluidextract of ..... 10 Strychnine = 1.05 to 1.25

(n.l.t.) = not less tl1an.

Questions and Problems

1. Why must care be. used to prevent emulsificaLion when shaking out
the ether-chloroform mixture. with i1cid solution?
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 429
2. Explain the chemical changes whieh the strychnine undergoes during
the various steps of the assay process.
3. Why is the determination of the total alkaloids of nux yomiea not a
correct measure of the therapeutic value of the drug'?
4. If the total alkaloids of nux vomiea were knowl~ to consist or strychnine
and brucine in the ratio 3: 1, what would be thc proper equivalent to use in
their volumetric estimation?

Assay of Caffeine-containing Drugs.-Caffeine is the ehief

alkaloidal constituent of a number of drugs used in different parts
of the world for beverage purposes such as tea, mate, coffel',
kola, and guarana, the last two of which are official. In the
determination of the caffeine content of the official drugs, it
should be borne in mind that caffeine differs in its properties
from most other alkaloids in the following ways: (1) It is very
feebly basie and cannot be determined by the volumetric method.
(2) The free alkaloid is appreciably soluble in warm water
(1 Gm. is soluble in 5.5 cc. at 800.). (3) It can be shaken out of
acid solution readily with chloroform. (4) It is not. precipitated
from solution by mercuric potl'tssium iodide. (5) It imblimes
without decomposition at about 180C.
The assays of guarana and kola are performed by the same
method.
Exercise 126

Object.-Assay of Guarana.
Materials Required.-6 Gm. of guarana in No. GO powder.
250 cc. of chloroform.
Procedure.~l. "Place G Gm. of Gu!tranll., in fine powder and accurately
weighed, into a suitable container, add 120 ce. of chloroform, allow the
mixture to stand about five minutes, and then add G cc. of ammonia Wl1ter
and 6 cc. of distilled Wl1tcr. Shake the mixture continuously for one llOur
or intermittently during two hours, and allow it to stRnd overnight.
Agl1in shake intermittently during one half hour and sepl1rate 100 ec. of
the clear chloroformic solution." .

That portion of the caffeine which occurs as a salt of tannic

acid is liberated by the ammonia and is dissolved in the chloro~
form along with the caffeine which occurs in the free state.
2. "Evaporate the chloroform, and treat thercsiduc with 10 cc. of approxi-
mately 1 pel' cent sulfuric acid with the aid of ~, gcntle heat. Filter the
cooled solution into a serll,rator.- and w'l.sb. the dish and filter with successive
4:10 QUANTITA'l'IVE PHARMACEUTICAL CHEMISTRY
~~

small portions of distilled water until no t.est for alk!1loid is obt.ained with
iodine T.S. in a portion (1 cc.) of the filtrate after acidulating strongly with
diluted sulfuric acid."

The aliquot portion of the chloroformic solution is evaporated

to dryness so that the alkaloid may be dissolved in acid. This
end is very difficult to accomplish by shaking out the chloroform
solution with acid since chloroform removes caffeine from acid
solution. The residue should not be dried at a tempera-
ture exceeding lOooe. because caffeine sublimes at higher
temperatures.
When the residue consisting of fat, coloring matter, etc., and
free alkaloid is treated with dilute sulfuric acid, the alkaloidnJ
sulfate dissolves in the aqueous solution, leaving the water-
insoluble impurities. Caffeine is precipitated from an acidulated,
but not from a neutral or alkaline aqueous solution by iodine
test solution.
3. "Make the combined filtrat,es alkt1linc with ammonia water and again
extract the alkaloid completely with successive portions of chloroform.
Evaporate .the combincd chloroformic solutions nearly to dryness, add about
1 cc. of neutral alcohol, and continue evaporation to dryness. Dry the
residue to constant weight at lOOC.
"The weight obtained represents the yield of anhydrous caffeine, Cal-Iw .
O.N., from 5 Gm. of Guarana."
The alkaloid liberated from its sulfate by the ammonia is
dissolved in the chloroform. Caffeine is determined gravimetri-
cally, since it is a very feeble base which docs not give a distinct
end point with any indicator upon titration.
The fiuidextracts of kola and guarana are made alkaline with
ammonia and extracted directly with chloroform. The dried
residue from the chloroformic solution is then treated with warm
dilute acid to dissolve the free alkaloid and the alkaloid is
shaken out from the aqueous solution with chloroform and
determined gravimetrically in the same way as in the assay of
the crude drug.
Questions and Problems
1. Why is not the caffeine extracted from the chloroformic solution with
dilute Rcid in the usual manner?
2. Could mercuric potassium iodide solution be used to test for the eom-
plete extractiOIl of the caffeine from the acid solution?
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 431
3. Why should eaffeine he dried at a low t.emperature?
4. Explain why warm water can be used to dissolve the frco alkaloid from
the ehloroformie residue in the U88fLY of the fluidextrfLct of kola.

TABLE LXVI.-Ol'FICIAI, DnUGs ANn GALENICAl, PREI'ARA'rlONS ASHAYEil

FOR CAFFEINE

Amount
use!l, Official requirement,
SubstmJCc
GIll. or per cent l'lLffeine
ce.

N.F.
Guarana .................. . o 4.0
GWLrana, fluidextract of ... . ii 3.6 to ,lAW/17
Kola .................... . 12 1.0
Kola, fluidextract of ....... . 10 0.85 to l.l5W /1'

Assay of Alkaloidal Salts.-The determination of the amount

of alkaloid present in an alkaloidal salt or in a simple preparation
containing an alkaloid or an alkaloidal salt is generally a relatively
simple process since the alkaloid does not need to be freed from
a number of organic impurities as is the case in the assay of crude
drugs. The solvents used and the gravinwtric or volumetric
methods employed uiffer according to the characteristic proper-
ties of the alkaloid determined.

Exercise 126

Object.-Assay of Citrated Caffeine.

Materials Required.-l Gm. of citrn.ted caffeine.
75 cc. of chloroform.
Procedure.-" Accurately weigh about 1 Gm. of Citrated Caffeine, pre-
viously dried to constant weight at SOC., and dissolve it in 10 cc. of hot,
distilled water; add 8 cc. of sodium hydroxide T.S., eool the solution, and
shake it in a separaLor with three or more successive portions of 20 cc. each
of chloroform, to effect complete extraction. Filter the combined chloro-
formic solutions through a sUlall filter, previously moistened with chloroform,
into a tared porcelain dish. Wash the filter and funnel with 10 ce. of hot
chloroform, adding the rinsings to the dish, and evaporate the combined
chloroformic solutions on a water hath, adding 2 cc. of alcohol just before the
last trace of chloroform is expelled. Complete the evaporation of the
solvent and dry the residue, consisting of anhydrous caffeine, to constant
weight at 800."
432 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The citrated caffeine is dried before it is weighed to drive off

moisture. Since caffeine is a very weak base and citric acid is
feebly acidic, it is probable that the aqueous solution of citrated
caffeine is dissociated almost completely into caffeine and citric
acid. Citric acid is quite soluble in chloroform, but its sodium
salt is insoluble in this solvent. The chloroform, thorefore, dis-
solves the caffeine, leaving the sodium citrate in the aqueous
solution.
Exercise 127
Object.-Assayof Eucaine Hydrochloride.
Materials Required.--0.5 Gm. of eucaine hydrochloride.
100 ce. of neutrall1lcohol.
Procedure.-Dissolve about 0.5 Gm. of eucaine hydrochloride, pre-
viously dried to constant weight -at 1000. and accurately weighed, in
100 ce. of neutral alcohol. Titrate this alcoholic Rolution with 0.1 N sodium
hydroxide, using phenolphthalein T.S. as indiC[ttor. Each cubic centimeter
of 0.1 N sodium hydroxide corresponds to 0.02836 Gm. of O,H 7 N(OH a),-
O(OoH,OO).HCl.

Neutral alcohol is specified because the ordinary eommereial

alcohol frequently contains considerable amounts of acetic acid
which, if present, would react with the standard alkali. The
standard alkali solution reacts quantitatively with the alkaloidal
salt liberating eucaine:
CsH7N(CH3)aO(CaH5CO).HCI + NaOH~N[1CI + H 0 + 2
283.64
C~H7N(CH3)aO(C6H6CO)

The liberated alkaloid is dissolved in the alcohol. Each cubic

centimeter of 0.1 N NaOH corresponds to 10 2~31~~00 = 0.02836
Gm. eucaine hydrochloride.
E:xercise 128

Object.-Assay of Theobromine with Sodium Salicylate for

Theobromine.
Materials Required.-2 Gm. of theobromine with sodillm salicylate dried
to constant weight at 1100.
Procedure.-l. "Dissolve about 2 GIll. of Theobromine with Sodium
Salicylate, previously dried to constant weight at nooO. and accurately
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 433

weighed, in 10 cc. of warm distilled water, and titrate the solution with
normal hydrochloric acid, using phenolphthalein T.S. as the indieator: not
more than 5.5 cc. of normal hydrochloric acid is required to neutralize 2 Gm.
of the (lried Theobromine with Sodium Salicylate."

Sodium theobromine is an unstable compound which is formed

when the alkaloid is dissolved in a solution of f;odium hydroxide.
A solution of sodium theobromine is strongly alkaline, and
the alkali may be' titrated directly with standard acid, the theo-
bromine being precipitated at the same time. In the procpss
of manufacture of theobromine with sodium salicylate, a slight
excess of alkali is added. If only the theoretical (J_uantity of
alkali were used, sufficient carbon dioxide would be absorbed
during the powdering and drying to make the product incom-
pletely soluble in water. A limit of titratable alkali is necessary
to prevent the use of products containing too large an excess.
In the titration, only the sodium combined with the theobromine
and that in exceSR is determined sinc:e the titration is stopped
at the phenolphthalein end point whieh is on the alkaline side
of the pH scale. The sodium salicylate doefl not enter into the
reaction.
2. "Thc solution should now he slightly alkaline to litmus paper or be
made so by the addition of 1 or 2 drops of very dilute fllmnonis. T.S. Allow
to stand at from 20 to 25C. for three hours, stirring occasionally, transfer
the precipitate of theobromine to a dried and weighed filter of 9 em. dhtmeter,
wash the precipitate !Illd filter with fnur successive portions of 5 cc. each
of ice-cold distilled water, find elr)' to constHut weight at 100C. To the
weight of precipitate thus obt.ained add 0.13 Gm. (the approximate quantity
of theobromine remaining in the liquid and washings). The total weight
corresponds to not less than 46.5 per cent of the weight of dried Theobromine
with Sodium Salicylate taken for the assay."

The theobromi.ne is precipitated from the alkaline solution,

collected, washed, dried, and weighed. Sinee theobromine is
slightly soluble in the liquid from which it is precipitated, a C01'-
r'3ction is applied to the result.
It is difficult to separate the theobromine from the mixture
because acidification liberates both the theobromine and the
salicylic acid and both of these compounds form soluble products
with excess of alkali. Theobromine is practically insoluble in all
of the common organic solvents and therefore cannot be extracted
434 QUANTITATIl'E PHARMACEUTICAL CHEMISTRY

in the usual manner. A better method of assay of the theo-

bromine with "odium salicylate than the official method is one
which is based upon the conversion of the theobromine, 3: 7
dimethyl-xanthine, into caffeine, 1: 3: 7 trimethyl-xanthine, by
a procedure similar to that given for the assay of theophylline
with sodium ncetate.

Exercise 12.9

Object.-Assay of Theophylline with Sodium Acetate for

Theophylline.
Materials Required.-l Gm. of theophylline with sodium acet.ate.
0.8 ce. of dimethyl sulfatc.
60 ce. of chloroform.
Procedure.-l. "Disperse about 1 GIll. of Theophylline with Sodium
Acetate, accurately weighed, in 5 ce. of distilled water amI '1.5 ee. of normal
sodium hydroxide, in a small stoppered fillSk. Add 0.8 ce. of dimethyl
sulfate, shake the. mixture until solution is complete, [tile! set it aside for one
hour, shaking it occasioIHtlly. Then add 5 cc. of Ilurmal sodium hydroxide
and 10 ceo of distilled waLer and shake the lllixture for one minute."

The theophylline, 1: 3 dimethyl-xanthine is converted quantita-

tively by methylation into eaffeinc, 1: 3: 7 trimcthyl-xanthille,
when the dissolved sample is shaken with dimethyl sulfate in
the presence of sodium hydroxide;

2. "Transfer it to a separator with the llid of chloroform and a little

distilled wa~er, a.nd extract the caffeine thus formed immediately by shaking
the liquid successively with 10-, 10-, 10- !Lnd 5-ce. portions of (~hloroform,
passing each chloroformio solution through It SIll!]Jl filter, previously moist-
ened with chloroform, into a tared poreclain dish, finally washing the stem
of the separator !Lllcl the filter with 10 cC. of hot chloroform. Evapomte the
combined chloroformie solutions on f\ wn,ter bath, adding 2 ce. of alcohol
before the last traces of chloroform arc expelled. Complete the cva.poration
of the solvent, and dry the residue, consisting of anhydrous caffeine, to
constant weight at 80C,: the weight of anhydrous caffeine so oMained,
multiplied by 0.9287, indicates the corresponding weight of anhydrous
theophylline, "

The caffeine formed, unlike theophylline, does not form a

soluble product with alkali and can be extracted with chloroform.
 ALKALOIDAL ASSAYS BY SPECIAL METHODS 435

Exercise 130

Object.-Assay of Ephedrine Sulfate.

Materials Required.--O.5 Gm. of ephedrine sulfate.
100 ee. of ether.
10 CC. of alcohol (neutralized to bromthymol hllle).
Procedure.-l. "Transfer about 0.5 Gm. of Epho(irinc Sulfat.e, dried oyer
sulfuric acid for twenty-four hours and accurately weighed, to a seprll'ator;
add 10 ce. of distilled water and 5 ce. of ammonia T.8., and extract with six
portions of ether, using 25 cc., 20 ce., 15 ce., 15 ee., 15 ce., and 15 ce ..
respectively. "

The ephedrine liberated from the sulfate hy the ammonia is

extracted by the ether. Chloroform cannot be mlcd to shake out
the ephedrine because this alkaloid is a sufficiently strong base to
decornpose the chlorofor111 and form ephedrine hydrochloride
especially in warm solution. (See test for identity of ephedrine)
U.S.P., page 144.)
2. "Collect the extracts in a beaker, evaporate the ether on a water bath
to about 5 cc., add 10 ce. of alcohol, previollsly neutralized to bromthyn101
blue T.S. with tenth-normal sodium hydroxide, and add from a burette an
excess of tenth-normal hydrochloric acid. Titrate the excess of acili with
tenth-normal sodium hydroxide, using bromthymol blue T.S. as the indi-
cator. The ephedrine thus found is not less than 75.5 and not more than
77.3 per cent of the weight of Ephedrine Sulfate taken. One cubic centi-
meter of tenth-normal hydrochloric acid is equivalent to 0.016.51 Gm. of
ephedrine."

The extracted ephedrine is dissolved by the alcohol and con-

verted quantitatively to ephedrine hydrochloride by the acid:
C1oH1SON + HCl-?C 1oH 16 0N.HCl
165.13 201.60
436 qUANTITATIVE PHARMACEiJ1'ICAL CHEMISTRY
TAIlLE LXVII.-ASSAYS OF ALKALOIDS, Ar,KALOIDAL SAW'S, AND SIMPLE
PREPARATIONS
Amonnt
used,
SubHtnllC'.C Gm. or
ce.
'T.S.P.
C~\:ffuine, dtrllted ........... .... . 1 Anhydrous caffeine = 48 to G2
Caffeine with sodium benzol1te ... . 1 Anhydrous caffdlle = 47 to 50
Codeine phosphate ............. . 0.5
Ephedrine ..................... . 0.5
Ephedrine hydl'tlchloride ........ . 0.5
Ephedrine sulfatc ............... . 0.5
l%hylhydrocupreine hydrochloride 0.5
Eucaine bydroehloride ......... , . 0.5
Phenacaine hydrochloride ....... . 0.5
Quinine l1ud nrea hydrochloride .. . 0.5
Theobromine with sodiurfi salicy-
late ......................... . 2 Theobromine = (n.1.t.) 46.1i
Theophylline with ethylene dia-
mine ........................ .
Theophylline with sodium acctate.
N.F. .
Apomorphine hydrochloride, tab-
lcts 01. ................... 0.00" Apomorphine hydroohloride = 0] to 10()b
Atropine suUate, tablets of
containing 0 . 02 or more Gm .... 0.08a
containing 0.0012 to 0.02 Gm . 0.08"
containing less than (}. 0012 Gm. 0.08"
Cnffdnc citratcd, tablets of .. ' ... 0.5"
Caffeine with sodium benzoate,
nmpuls of ................... . 0.5"
Caffeine with sodium benzoate,
tablet" of .................... . 0.5a
Cocaine hydrochloric!", to.blets of .. 0.06 1
Codeinc phosplmte. tablets of ..... 0.06"
Codeine Bulfnte. tablets of. ...... 0.06"
Emetine hyrlrochloride, ampuls of 0.3 a
Ephedrine hydrochloride. tablcts of 0.06"
Ephedrine sulfate, ampul" of ..... . 0.5 a
Ephedrine sulfate, solution of .... . 10
Morphine sulfate, tablets of ..... 0.13"
Morphine and atropine Bullates,
tablets of. ................... 0.13"
Nux vOlnic[1 nlkaloids, solution of.. 20
Prooaine hydrochlorirle, ai:npuls of 0.2"
Procaine hydrochloride, solution of 10
Procaine hydrochloride, tablets of. 0.211
Quinine dihydrochloride, ampuls of 0,5 a
Quinine hyurochloride and ethyl
carbamate, am puIs of. I Quinine hydrochloride = 02.5 to 107.5 1,
Quinine and urea hydrochlorides,
ampuls of .................. O. fia
Quinine phosphate .............. . 0.5
Quinine sulfate, tablets of. ...... . 0.13a
Scopolamine hydrobromide, tablets
of ........................ 0.06 a
containing 0.0012 Gm. or more.
containing less than 0.0012 Gm.
Strychnine nitmte, tablets of .... 0.l3"
containing 0.02 Gm. or more ...
containing 0.0012 to 0.02 Gm ..
containing less than 0.0012 Gm.
Strychnine phosphate ...........
Strychnine sulfate. Lablets of ....
containing 0.02 Gm. or more ...
containing 0.0012 to 0.02 Gm ..
containing Jess than 0.0012 Gm.
(n.1,t.) = not leBS than.
~Amount of eqnivnlent Bougbt.
~ Per cent of the labeled amount.
Official I'cquireJnent, eent
PCI'
Anhydrous codeine = (n.l.t.) 70
Eph .. drine = 08 to 100
Anhydrous ephedrine = 80 to 82.5
Anhydrous ephedrine = 75.5 to 77.3
Ethylhyclrocupreine = (n.Lt.) 90
Eucaine hydrochloride = (n.1.t.) 9il
PheUl1cuine = 87.5 to DO. 5
Anhydrous quinine = 58 to Hf)
Anhydrous j.h~ophyllille = 70 to 80
Anhydrous theophylline = 65 to 65
Atropine sulfate = 02.5 to 107.!jb
AtrGpine sulfate = \11 to 10U/'
Atropine Aulf'Lte = 88 to 112"
Co.ffcine, citratcd = 43 to 53 b
Anhydrous caffeine ;=; 4[j to 52 b
Anhydl'ouB cn:ffeine 4a. [) to 53.511
=I
Cocaine hydroc:hlorid(, = 91 to 109b
Anhydrous codeine = 03 to 77 b
Codeine. sulfate .= 01 to 1m)l!
Emctinu.hydrochlodde = 84 to 92'
l,phcdrine hydrochloride = ()1 to 109'
Ephedrine = 72. Q to 80. 2b
Ephedrine Aulfat.e = 2.8 to 3 .2TV IV
Morphine sulfute = 92.5 to 107.5'
Morphine sulfate = 9 1 to 100 b
Str:l'ehnine sulfate = 1. fi to 1. 7TV IV
Brucine sulfate = 1.5 to 1. 7W IV
Prooaine hydrOl!hlorifi! = Uri to l05 b
Procaine hydrochloride = 1.0 to 2.1 TV IV
Procaine hydrochlori(ip- = 92.5 to 107.5 11
Quinine dihydrocblol'ide = 05 to 105 b
Quinine and urea llydl'oclllOl'idoB = 54.8 to
63.7 b
AnhydrouB quinine = 74 to 78
Quinine sulfa.e = 01 to 10gb
Scopolamine hydrobromide = 91 to 10gb
Scopolamine hyurobromi(l" = 88 to 112b
St.rycbnine nitmte = 92.5 t.o l07.5b
Strychnine nitratE! = 91 to 109b
Strychninc nit.rate = 88 to 112b
Strychnine = 70 to 73
Strychnine sulfate = 92.5 to 107.5b
Strychnine sulfnte = 91 to 10g b
Strychnine sulfate = 88 to 112b
 CHAPTER XXVII

OTHER OFFICIAL ASSAYS INVOLVING THE USE OF

IMMISCIBLE SOLVENTS
Use is made of the immiscible solvent principle of extraction
and purification in the proximate determination of the amount
of active constituent contained in a number of non-alkaloidal
drugs such as jalap, cantharides, and oleoresin of aspidium and
in the assay of certain synthetic medicinal products, e.g., soluble
barbital.
Exercise 181
Object.-Assay of Cantharides.
Materials Required.-15 Gm. of cantharides in No. 40 powder.
75 ce. of purified petroleum benzin.
100 ce. of benzene.
50 cc. of chloroform.
25 cc. of absolute alcohol.
Procedure.-l. "Place 15 Gm. of Cantharides, in moderately coarse
powder, in a pressure bottle of not less than 250 cc. capacity, add 150 cc. of a
mixture of benzene, two volumes, and purified petroleum benzin one volume,
and then add 2 co. of hydroehloric acid. Stopper the bottle tightly, shake
it well, and allow it to stand about ten hours. Now gradually warm the
bottle and its contents to about 40C. and maintain it at approximately
that temperature with frequent shaking during three hours, avoiding
evaporation. If necessary, add more solvent to replace any lost by
evapomtion."

Cantharidin, C SH 12 0:(COh:O, is the inner anhydride, lactone,

of the dibasic cantharidie acid, C SH 12 0(COOH)2_ Both can-
tharidin and the alkali salts of calitharidic acid occur in the
beetles. When the above mixture is acidulated, the cantharidin
and the liberated eantharidic acid dissolve in the warm benzene-
petroleum benzin mixture together with fats, coloring matter,
and other inactive constituents. Although a Soxhlet apparatus
can be used to advantage in this assay, the official directions are
to extract the drug by maceration, the purpose -being to adapt
437
438 QUANTI'l'A'l'IVE PHARMACEUTICAL CHEMISTRY

the method to the equipment avn,ilable in most pharmacputi(Jal

laboratories. The maceration should be carried out in a heavy
walled bottle (a magnesium citrate bottle serves very well),
since the volatile solvents genemte considerable pressure when
warmed to 40C.
2. "Cool the mixture, decant or filter off 100 ce. of the clear soll1t.ion, awl
evapomte this rapidly in a tared beaker or wide-necked flask to II volume
of about 5 cc. Add 5 cc. of chloroform to the residue and set it aside in a
moderately warm place."

The evaporation of the volatile solvents should be performed

as rapidly as possible) because cantharidin begins to sublime at
86C. and appreciable amounts are driven off with the Holvellt
during slow evaporation. When chloroform is added and the
mixture is allowed to stand in a warm place, the cantharidin
crystallizes out upon evapoJ,'ation of the chloroformic solvent.
3. "When the solvent has all evaporated, add to the crystals 10 ec. of
a mixture of equal volumes of dehydrated alcohol and pm'ified pctJ'Oleul1l
benzin, which has previously been sttturated wit.h pure cantharidin, allow
the mixture to stand fifteen minutes, and thon dccnllt tho liquid through
a pledget of purified cot,ton. Wnsh the crystals with successive portions
of !j, saturated solution of cantharidin, similnr to that dirm~t(Jd above, until
free from fat and coloring matter, and pass the washings through the same
pledget of purified cotton."

Cantharidin is only slightly soluble in the purified petroleum

benzin and dehydrated alcohol mixture. The use of a mixture
of these solvents previously saturated with cantharidin makes it
possible to remove the fat, coloring matter, etc., with which the
crystals are contaminated without dissolving any of the crystals.
Successive 2 to 3 cc. portions of the above wash mixture serve
most efficiently to remove the impurities adhering to the crystals.
4. "Then wash the cotton with II very SIllltll quantity of warm chloroform
to dissolve any adhering crystals, collect the chloroform in the tared flask
or beaker containing the washed crystals, evaporatc the solvent with the
aid of a current of ail', dry the Cl'yst:l1s at 600. for one-half hour, and weigh.
The resulting weight will be the amount of cantharidin obtained from
10 Gm. of Cantharides."

Negligible quantities of the cantharidin are volatilized when

it is dried for one-half hr. at 60C. The residue, which consists of
cantharidin and cantharidic acid in variable amounts, is con-
ASSAYS INVOLVING 'J'HE USE OF IMMISCIBLE SOLVENTS 439

sidered to be cantharidin. The weight of the residue corresponds

to the cantharidin contained in 10 Gm. of the sample assayed.
The U.S.P. requires that cantharides yield not less than O.B
per cent of cantharidin when assayed as directed above. Cal-
culate the percentage of cantharidin contained in the sample
assayed and eompare the results with the official requirement.
Questions and Problems
1. Why is hydrochloric acid added in t.hc initial ext.ruction of the
cantharides?
2. Why should the residue be dried at, II low temperature?
3. Why is the maceration conclUded with a warm solvent in 11 llw;.;sure
bottle?

Exercise 132

Object.-Assay of Oleoresin of Aspidium.

Materials Required.-About 3 Gm. of oleoresin of aspidium.
150 ee. of ether.
3 Gm. of barium hydroxide.
Procedure.-l. "Warm the Oleoresin on 11 wat.er hath and stir unm it is
thoroughly mixed. Transfer about 3 Gm. of it, accurately weighed, to a
250-cc. flask, dissolve it in 40 ee. of et,her, add 75 ce. of a 3 PPl' cent aqueous
solution of barium hydroxide, and sh!lke the mixture vigorously for five
minutes. Transfer the mixture to a separator, anow thc liquids to separate
completely, and dmw off and filter the barium hydroxide layer. Rinse the
250-cc. flask with two 25-ec. portions of a 3 per cent aqueous hariuIll hydrox-
ide solution. After each rinsing, tmnsfer the barium hydroxide solutiCln
to the sepltrator, slutke the mixture for one minute, allow the liquids to
separ9.te completely, and draw off and filter the bariuJIl hydroxide layer."

Oleoresin of aspidium usually deposits granular, crystalline

filicic aeid or filiein upon standing. It is warmed to render it
morn fluid so that the precipitate may be uniformly distributed
throughout the sample. The filmarone originally present, in
the crude drug, aspidium, is convertecllargely into filicic aeid and
its amorphous modification, filicill, during the preparation of the
oleoresin. The barium hydroxide neutralizes the filmarone,
filicic acid, and filicin, forming the water-soluble barium com-
pounds which dissolve in the aqueous layer of the immiscible
solvents. The resin, fat, coloring matter, etc., remain dissolved
in the ethereal layer.
440 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

2. "Transfer the combined filtered barium hydl'ox~dc solutions to a

separator, render distinctly acid to litmus paper by the !Lddition of hydro-
chloric [wid, and extract with three successive portions of 30 ee., 20 ee., and
15 ce. of ether."
The filicic acid and filicin are liberated from their barium salts
and dissolved in the ether, leaving barium chloride in the aqueous
solution.
3. "Filter the combined ethereal solutions, wash the filter with ether,
evaporate, and dry the residue to constant weight at 1000. This residue
is calculated as crude filicin and its weight should be not less than 24 per
cent of the weight of Oleoresin tilken for the assay."

The residue consists mostly of filicin and small amounts of

filicic acid.
Calculate the percentage of crude filicin contained in the
oleoresin of aspidium., assayed, and eompare the result with the
official requirements.
Questions and Problems
1. Why should the oleoresin be warmed and mixed well before the sample
is taken?
2. Why is the sfLlnple of oleoresiu dissolvod in ether? WIULt function
does the ether serve?
3. Look up the assay method for aspidium in the U.S.P. and expla.in
briefly how this crude drug is assnyed.

Exercise 133

Object.-Assayof Jalap.
Materials Required.-lO Gm. of jalap in No. 60 powder.
20 ce. of 1 per cent hydrochloric acid. .
90 ce. of chloroform.
120 ce. of alcohol.
Procedure.-l. Place 10 Gm. of jalap, in fine powder and accurat,ely
weighed, with about 60 cc. of a mixture of 9 volumes of alcohol and 1 volume
of water, in a flask provided with a reflux tube or condenser, and digest the
mixture on ll. steam bath during Y:l hr. 'rransfer the warm mixture to a
small pcrcolntor, allow i~ to drain, press the marc down gently, and percolate
with tho warm alcohol-water mixture until 100 cc. of percolate, whon cooled,
is obtained, and mix thoroughly. .

The jalap is digested and percolated with alcohol to extract

the resin completely,
ASSAYS INVOLVING THE USE OF IMMISCIBLE SOLVENTS 441
2. Pipet,te 20 cc. of this percolatc into a separator, arid 40 ce. of ~hlorofDrm
and 10 cc. of approximately 1 per cent hydrochloric acid, and shake for
2 min. Allow the mixture to separate, draw off the chloroformic l:tyer, and
extract the aqueous layer twice more, using 15 ce. of a mixture of 2 volumes
of chloroform and 1 volume of alcohol each time. Shake thc combined
chloroformic solutions with 10 cc. of 1 per cent hydrochloric acid, again
separat.e, and wash this acid liquid twice with 15 ce. portions of chloro-
form-alcohoL

The resin of jalap is insoluble in aqueous acid solution. It

dissolves in the alcohol-chloroform layer leaving nOll-resinous
constituents in the aqueous layer.
3. Pass the combined chloroformic extractions through filter paper moist-
ened with the chloroform-alcohol mixture, wash the filter with the chloro-
form-alcohol mixture, and cvapomte the filtmtc to dryness; lLdd 2 cc. of
alcohol and again evaporate. Dry the residue to constant weight at 1000.
The weight obtained represents the yield of total resins from 2 Gm. of
jalap.

The lower aqueous layer containing small amounts of sugars,

gums, coloring matt.er, etc., is drawn off and discarded. The
alcohol-chloroform solut.ion when evaporated to const.ant weight
leaves a residue which consist.s of practically pure resin of jalap.
Ipomoea and podophyllum are also assayed by analogous
methods, since t.he solubility properties of the resins which they
contain are similar to those of resin of jalap.
Exercise 134
Object.-Assay of Tablets of Phenobarbital.
Materials Required.-20 tablets of phenobarbital.
10 ce. of a 2 per cent solution of NaOH saturatee- with NaOl.
About 20 cc. of alcohol.
About 50 cc. of ether.
About 75 ce. of chloroform.
Procedure.-l. "'Veigh not less t,han 20 of the Tablets, and reduce them
to a fine powder without an appreciable loss. Transfer an aliqnot portion,
equivalent to about 0.3 Gm. of phenoblubital, to a separatoI'}' funnel, and
dissolve it, as far as possible, in 10 CC. of ~Ln alkaline salt solution made by
saturating an aqueous 2 per ,cent SOdiUIll hydroxide solution with sodium
chloride. Wash the solution with two 15 ce. portions of ether and discard
the washings."

The phenobarbital reacts with the NaOH to form soluble

phenobarbital. When the solution is shaken with portions of
442 QUl1NTI1'A'l'IVE PHARMACEU'l'ICA.L CHEMISTRY

ether, organie matter, sueh as petrolatum used as a lubrieant in

the preparation of the tablets, dissolves in the ether but the solu-
ble phenobarbital being insoluble in ether remains in the aqueous
layer.
2. "Add 2 cc. of hydrochloric acid and 5 cc. of distilled water, and com-
pletely extract tho phenobarbital with a solvcnt composed of two volumes
of tllcohol, one volume of ether, and seven volumes of chloroform. Wash
the extract, with 5 ec. of distilled water, acidified with a drop of hydrochloric
llcid. Filter the extract through a pledgct of cotton into a tared bellker,

TABLE LXVIII.-OFFWIAL NON-ALKALOIDAL SUBSTANCES ASSAYED BY

METHODS EMPLOYING IMMISCIBLE SOLVENTS

Amount
tlsecL
Substunee Official requirement, pcr cent
Gm. or
oe.

U.S.P.
A8pidium., ...... , ........... ,., ..... . 125 Crude filicln =r 1.5
Aspidium. oleoresin of .. ............... . 3 Crude filicin = 24
Cantharides .................... , .... . 15 Cantharidin = 0.6
Podophyllum ........................ . 10 Resin = 4
Soluble barbital. .. , ..... , ... , ...... , .. Ba,'bital = 88 to liD
Soluble phenobarbital. .......... , . , . , .. 0.5 Phenobarbital = 90.4 to 91. 4
N.F.
Acetophenetidin, tablets of. .. , . , , .... , . 0.3" Acetophenetidin = 92,5 to 107. 5 b
Acetophenetidin and phenyl sltlicylate,
tablets of.,. , ...................... .
Aminopyrine elixir of . ................ .
t 5 Aminopyrine = 3.7 to 4.3W/V
Aminopyrine, tablet. of .............. , , la Aminopyrine = 92.5 to 107.5'
Barbital, elixir of ..................... . 10 Barbita] = 3.2 to 3.8W/V
Barbital. tablets of, more tlmll 0.07 Gm .. 0.3 a Barbit"J = 92.5 to 107.5'
0.07 Gm. or less ................... . 0.3" Barbital = 91 to 109'
Ipomoea, ........................... . 10 Total ro.in" = 15
Jalap., ... ' ....... , .... ", .. " ....... . 10 Totul resins = 9
.Talap, fluidextract of. , .. , . , .. , ........ . 2 nesin = 8.5 to 9.5W/V
.Talap, tincture of ....... , ........... , .. 10 nesin = 1.7 to 1.9W/V
Phenobarbital, elixir of. ...... , . , ...... . 25 Phenobarbital = 0.38 to 0.42TV/V
Phenobarbital, tablets of
more than 0.07 Gm ....... , ........ . 0.3" Phenobarbital = 92.5 to 107. 5b
0.07 Gm. or leB . , ........... , ..... . 0.3" Phenobarbital = 91 to laO'
Soluble barbital, tablet" of
more than 0.07 Gm ...... , ..... , ... . 0.3" Barbitnl = 02.5 to 107.5'
0.07 Gm. or lesB., .......... " ..... . 0.3" Barbital = 91 to 109 b
Soluble phenobarbital, t.ablets of
more than 0.07 Gm ..... , ...... ' ... . 0.3" Pitenobnrbitttl = 92.5 to 107. 5 b
0.07 Gm. or less ....... , ........... , 0.3 a Phenobarbital = 91 to 101lb

a Amount of ingredient sought.

, Per cent o( the labeled amount.
ASSAYS INVOLVING THE USE OF' IMMISCIBLE SOLVENTS 443

and wash tho cotton with small portions of the solvent. Evaporate the
oombined filtrate and washings 011 a water bath with the aid of a current of
air, and dry the residue of phenobarbital to constant weight at a temperature
not exceeding 100C."

The HClliberates the phenobarbital from the soluble pheno-

barbital and the liberated phenobarbital is dissolved and extracted
upon shaking with successive portions of the mixed orgtlnic
solvents. .
Questions and Problems
1. Calculate the amount of phenobarbital in each of the tn.blets.
2. Does the amount of phenobarbital found fall within the tolerance
limits established for these tablets in the National Formulary?
3. Consult the U.S. Phannacopoait1 for the formullla of phenobarbital
fmd soluble phenobarbital and write equations for the reactions tlmt occur
in the assay using graphic formulae.
 OHAPTER XXVIII

ASSAY OF ENZYME-CONTAINING SUBSTANCES

No methods are known whereby the quantity of enzymes in a
substance can be accurately determined. Consequently,' the
assay of enzyme-containing substances is based on obserVlttions
of their power to produce changes in other substances. In
other words, the activity of the enzymes is measured. Thus,
pepsin is evaluated on the basis of the quantity of egg albumen
which it will digest, and extract of malt is assayed for its power to
convert starch into soluble carbohydrates under definite condi-
tions. Any condition which tends to increase or decrease the
activity of the enzymes manifests itself in the assay results as
an increase or decrease in the quantity of onzymes present.
Numerous factors tend to influence the activity of enzymes.
Some of the more important of these factors are as follows:
1. Practically all enzymes are destroyed when they are heated
to 1000. Many of them are gradually inactivated, at com-
paratively low temperatures (35 to 40C.), and many of them
undergo a slow loss of activity at room temperature and even at
00.
2. Enzymes have an optimum temperature of action; that is,
for each enzyme there is a definite range of temperature at
which it exhibits its maximum activity under definite conditions.
The optimum temperature for most enzymes lies between 30 and
450.
3. The activity of the enzymes -is affected by the media in
which they react; e.g., pepsin is most active in acid media, and
pancreatin is most active in alkaline media.
4. The enzymes are inactivated by many chemical agents,
such as alcohol, salts of the heavy metals, tannin, and phospho-
tungstic acid, whic;h precipitate the enzymes from solution.
Because the activity of enzymes is influenced by so many
factors, it is necessary to adhere strictly to the procedures out~
444
 ASSAY OF ENZYME-CONTAINING SUBSTANCES 445

lined in the various assay processes in order to secure concordant

results.

Exercise 135

Object.-Assay of Pepsin.
Materials Required.--O.1 Gm. of pepsin.
0.1 Om. of U.S.P. reference pepsin. The U.S.P. reference pepsin consists
of a powdered pepsin, carofully selected, dried, and packaged and having It
dip;l1stive power of 3,000 times its weight of egg albumen.
Normal hydrochloric acid.
A hen's egg.
A No. 40 sieve.
A conical measure.
Procedure.-l. "Mix 35 cc. of normal hydrocllloric aeid with 385 ce. of
distilled water. Dissolve 0.1 Gm. of Pepsin in 150 cc. of this dilute acid.
Likewise dissolve 0.1 Gm. of Refcrence Pepsin in another portion of 150 ce.
of this dilute acid."

When 35 cc. of normal HCI is diluted to 420 cc., the concentra-

tion of acid in the solution is reduced to approximately 0.3 per
eent. This is about the normal acidity of the stomach. Investi-
gation has shown tJlat pepsin is most active as an albumen diges-
tive in media containing this concentration of acid. Other
strongly dissociated acids which supply the same hydrogen ion
eoncentration as 0.3 pel' cent RCI might be used, since the diges-
tive activity of the pepsin is directly related to the hydrogen ion
concentration of the reaction mixture.
2. "Immerse one or more hen's eggs in boiling water during fifteen
minutes. Cool them rapidly to room temperature by immersion in cold
water, removc the shell and pellicle and all of the yolk imd at once rub the
albumen through a clean, dry, No. 40 sieve, rejecting the first portion that
passes through the sieve. Place 10 Gm. of the succeeding well-mixed por-
tioIl in ench of three wide-mouth bot,tles of about 100-00. capacity."

The egg is immersed in boiling water for 15 min. to thoroughly

coagulate the albumen. The albumen is coagulated so that its
digestion by the pepsin may be better observed. The coagulated
albumen is passed through a sieve to break it into small particles
80 that a greater surface area will be exposed to the digestive
action of the pepsin enzymes. The first portion of the albumen
that passes through the sieve is rejected, since it may contain
446 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

dust particles and traces of metallic compounds from the mate-

rials of the sieve.
3. "Immediately add 35 cc. of the dilute !Leic] at one time or in portions
lmd, by suitlLhle means, disintegrate thoroughly the particles of albumen.
Plane the bottles in a water bath at 52C. After the contents of the bottles
have reached that temperature, add exactly 5 cc. of the acidulated solution
of Pepsin to one bottle, 4.30 cc. of the saIlle solution aIltl 0.70 ce. of the dilute
acid to another bottle, and exactly 5 ec. of the acicluI:lted solution of Refer-
ence Pepsin to the third bottle. At once stopper the bottles securely, invert,
them tbree times, ami maintain them at a tempemt.ure of 52C. for two and
one-half hours, agitating the contents equally every ten minutes by inverting
the bottles once."

The aQ.id should be addcd at once, hecause the

albumen loses moisture on exposure to the air and
becomes tough and harel. The mixture is main-
tained at 520., since pepsin exerts its optimum
activity at this temperature. Exactly 5 ce. of the
pepsin solution being assayed and of the solution
of reference pepsin is added, because this quantity
FIG. 67.- introduces 1 part of pepsin into the mixture to each
Sedimenta-
tion COIlO. 3,000 parts of albumen. Thus 0.1 Gm. of pepsin
in 150 cc. gives 0.1 :150: :X:5. X = 0.003333 Gm.
pepsin in 5 cc. of the solution. 0.0033 X 3,000 = 9.999 (10
Gm.), the weight of albumen used.

4. "Remove the bottles from the bath, pour t,he contents into 100-cc.,
conically-sh!,ped measuring vessels, Fig. 67, having cli!lll1eters not. exceeding
1 em. at the bottom, and gradu!1ted from the tip to the 1.0-ee. mark in
0.05-ce. divisions and from the 1.0-ce. to the 5.0-ce. mark in O.l-ce. divisions,
and having the .internal taper as nearly identical as possible. Transfer the
undigested egg albumen which adheres to the sides of the bottles to the
respective measuring vessels with the aid of sll1!Lll portions of distilled water
until 50 ce. has been used for each. Mix the contents of each measuring
vessel and allow them to stand for thirty minutes. The volulIle of the
.undissolved albumen in the mO!LSllring vessel corresponding to 5.0 cc. of the
solution of Pepsin being assayed shall not be more than the volume of
the undissolved albumen in the measuring vessol corresponding to 5.0 cc.
of the Reference Pepsin solution, and the volume of the undissolved albumen
in the measnring vessel corresponding to 4.30 ce. of the solution of Pepsin
being assayed shall not be less than the volume of the undissolved albumen
in the measuring vessel corresponding to 5.0 cc. of the Reference Pepsin
solution." .
 ASSAY OF ENZYME-CONTAINING SUBSTANCES 447

If the volume of egg albumen remaining undisHolyecl from the

sample containing 5.0 cc. of the solution of pepsin being assayed
is not greater than that remaining from the sample containing
the solution of reference pepsin, it is evident that the p~)p::;in
being assayed lllust be at least as active as thp, reference pepsin
and that it must conform to the minilllum official reqnirement.
If the volume of egg albumen remaining undissolved from the
sample containing 4.3 cc. of the solution of pepsin being assltyed
is not less than that remaining from the sample containing the
reference pepsin! it is evident that the pepsin being assayed is
a pepsin which will cause the solution of 3,500 times its weight
of egg albumen. 4.3: 5: : 3,000: X. X = 3,500. The relative
proteolytic power of pepsin, stronger or weaker than that just
described, may be determined by ascertaining through repeated
trials the quantity of the pepsin solution made as diredecl in the
assay, required to digest under the prescribed conditions, 10 Gm.
of boiled and disintegrated egg albumen.

Questions and Problems

1. Name several factors that, affect the activity of pepsin.
2. Why is the proteolytic activity of pepsin determined in acid solution?
What effect would thc usc of a neutral or alkaline medium have on the diges-
tive power of pepsin?
3. Could proteins other than egg albumen be used to estimate the proteo-
lytic activity of pepsin? Give examples.
4. What advantages does egg albumen possess as a source of protein for
this assay?

Exercise 136
Object.-Assay of Pancreatin for Starch Digestive Power.
Materials Required.-5 Gm. of powdered pot,ato starch.
0.3 Gm. of pancreatin.
0.2 ec. of 0.1 N iodine solution.
Procedure.-l. "Determine the percentage of moisture in potato starch
by drying about 0.5 Gm., accurately weighed, at 120C., for four hours.
Thoroughly mix a quantity of the starch, equivalent to 3.75 Gm. of dry.
starch, with 10 cc. of cold distilled water. Add the mixture with constant
stirring to 75 ce. of distilled water, previously heated to from 50 to 60C.,
contained in a tared, 250-cc. beaker. Rinse the remaining starch into the
beaker with 10 cc. of distilled water, heat the mixture to boiling, and boil it
gently, with constant stirring, for five minutes, or until a translucent, uni-
form paste is obtained."
448 QUANTI'l'A'NVE PHARMAOEUTICAL CHEMISTRY

The percentage of moisture remaining in the starch is deter-

mined so that a sample equivalent to 3.75 Gm. of dry starch
can be used for digestion. Thus, if the sample dried at 50C.
is found to contain 10 per cent of moisture, each gram of the
sample is equivalent to 0.9 Gm. of dry starch. Then 3.75/0.9 =
4.1666 Gm. starch equivalent to 3.75 Gm. of dry starch.
The stareh is mixed with cold water to obtain a paste so that it
will not form lumpy masses when the hot water is added. When
this paste is added to warm water and the resulting mixture is
boiled, the starch grains burst, liberating their contents whieh
dhlsolve in the hot water to form a more or less mucilaginous
mass. Until the starch grains are broken down in this way,
they remain insoluble and are not acted upon by pancreatin.
2. "Add enough distilled water to make the mixture weigh 100 Gill.,
cool the paste to 40C., and place the beaker on a water ba1,h maintained
at 40C. Suspend 0.15 GIll. of Pancreatin in 5 ce. of distilled water and add
the suspension to the stal'ch paste, mixing it well by pouring the mixture
from beaker to beaker for thirty seconds, noting the time when the Pan-
creatin slIspension was added to the starch. Maintain the mixture at a
temperature of 40C. for exactly five minutes."

The water lost by evaporation is replaced so that a definite

concentration of starch will be present during the digestion with
the pancreatin. The solution of pancreatin must be prepared
immediately before it is added to the starch solution I because
amylopsin l the stareh-splitting enzyme of pancreatin, becomes
inactivated quite rapidly in aqueous solutions. The mixture is
maintained at 40C. for 5 min., since 40C. is the optimum
temperature for amylopsin. The regulation of the temperature
is very important in this assay, since a variation of 1C. through-
out the period of digestion has been found to cause a difference
of about 10 per cent in the activity of the !Lmylopsin of pancreatin.
The amylopsin converts the starch into soluble carbohydrates,
chiefly maltose and isomaltose.
3. "At once add 0.1 cc. of this mixtnre to a previously made mixture of
0.2 cc. of tenth-normal iodine and 60 ce. of distilled wtttel' ttt tt t,emperatul'e
of from 23 to 25C.: no blue, red 01' violet color is produced. JJ
The presence of starch and primary hydrolytic products of
starch is indicated by tho development of a blue, red, or violet
color.
 ASSAY OF ENZYME-CONTAINING SUBSTA.NCES 449

Pancreatin must be capable of cOIlverting at least twenty-five

times its weight of starch into soluble earbohyclratps in order
to meet the pharmacopoeial requirements. Since 0.15 Gm. of
pancreatin is employed to digest 3.75 Gm. of starch, each Gm. of
pancreatin used corresponds to 3.75/0.15 = 25 Om. starch. The
relative amyloptic activity of a Barnple of pal1(~reatin can he
determined by repeated trials on varying quantities of sta,reh.
Extract of malt of the U.S.P. is abo assayed for its capaeity to
convert starch into soluble carbohydrates which do not give a
color with iodine solution.
Questions and Problems
1. Define the terms proteolytic enzyme and mnylolytic el!zyme.
2. Why is the stl,reh solution boiled before the digestion with pancreatin?
8. Look up the hydrolytic products of starch and explain wllllt produets
might give 11 red or violet color with iodine?

EXercise 137
Object.-Assayof Pancreatin for Casein Digestive Power.
Materials Required.-0.1 GIll. of casein.
1 CC. of 0.1 N sodium hydroxide.
01. GIll. of pancreatiu.
1 ee. of glacial acetic acid.
10 cc. of alcohol.
Procedure.-1. "Place 0.1 GIll. of finely powdered casein ill!l 50 ce. volu-
metric flask, add 30 cc. of distilled water, and shake well to bring the casein
into suspension. Add exactly 1 cc. of tenth-normnl sodium hydroxide, Hud
heat the mixture at 40C. until the casein is completely dissolved, which
should not require more thnn thirf,y minutes. Cool, add sufficient distilled
water to make 50 ce., and mix well."

Casein is a complex compound, or mixture of eompounds,

which yields amino acids upon hydrolysis. Consequently, it is
soluble in alkalies. The casein is dissolved in alkali, in this ease,
because trypsin, the proteolytic enzyme contained in pancreatin,
functions best in alkaline media. Casein 'which requires more
than 30 min. to dissolve is usually of poor quality, containing an
excess of fat. '
2. "Dissolve 0.1 GIll. of Pancreatin in 500 cc. of distilled water. Mix 1 cc.
of glacial acetie neid with 9 ce. of distil1ed water and 10 ee. of alcohol. Place
5 ce. of the casein solution in a test tube, add to it 2 ce. of the well-shaken
Pancrel\tin solution and 3 cc. of distilled waier, and mix by gentle agitation.
450 QUAN'l'I7'ATIVE PHARMACEUTICAL CHEMISTRY

Immediately immerse the test tube in a water bath at 40C., and keep it
at this temperature for one hour. Then remove from the bath, and add 3
drops of the acetic acift mixture. No precipitate is produced."

The concentrations of the solutions of casein and pancreatin

are such thnt when thcy arc mixed there will be 25 partH of casein
present for each part of pancreatin. Thus, in 2 cc. of the pan-
creatin solution there is 2 X 500~ = 0.0004 Gm. pancreatin, and

in 5 cc. of casein solution there is 5 X ~.~ = 0.01 Gm. casein;

0.0004 X 25 = 0.0100.
The mixture is maintained at 40C., the optimum temperature
for trypsin, for 1 hr. to allow the proteolytic action to proceed.
The casein is digested by the pancreatic enzymes with the
formation of soluble proteoses and possibly small amounts of
amino acids. When the alcoholic acetic acid solution is added,
the alkali is neutralized and any undigested casein is precipi-
tated, since casein is insoluble in alcohol and in acetic acid.
The proteoscs and other products of casein digestion, being
soluble in alcohol and acetic acid, are not precipitated. Con-
sequently, if a precipitate is formed, it indicates that the pro-
teolytic activity of the pancreatin is below standard; that is,
that 1 part of the pancreatin is not capable of digesting 25 parts
of casein.
Questions and Problems
1. Why is the protein digestive capacity of panerelltin determined in an
alkilline medium?
2. Which of the enzymes of pancreatin take part in protein digestion?
8. Define the term optimum temlJ81'ature as applied to enzymes.
4. How might the relative proteolytie power of a sample of pancreatin
stronger or weaker than the official standard be determined?

Exercise 188
Object.-Assay of Rennin.
Materials Required.-O.lGm. of rennin.
0.1 Gm. N.F. reference rennin (a carefully preserved, stable, powdered
rennin that has been repeatedly tested for r number of years so that its
stability and its standard are definitely established. It Coagulates approxi.
mately but not less than 25,000 times its weight of fresh cow's milk).
100 cc. of cow's milk.
 ASSAY OF ENZYME-CONTAINING SUBSTANCES 451

Procedure.~l. "Mix 0.1 Gm. of Rennin with 50 ce. of distilled water by

gentle stirring, and allow the mixtu re to stand exactly 15 minutes. Lik~
wise, mix 0.1 Gm. of Reference Rennin (see page 450) with 50 ee. of distilled
water by gentle stirring, and allow the mixture to stand for exactly 15
minutes."

When rennin is added to water, it is taken up slowly, forming tL

somewhat opalescent mixture. If shaken vigorously, the rennin
tends to clump into large particles which separate out. When
the mixture is stirred gently and allowed to stand, however, the
rennin gradually absorbs water and dissolves.
2. "Place 50 ee. of well-mixed (lOW'S milk in eaeh of two glass vcssels
about 12 em. high and 4.5 em. in diameter. Warm the milk rapidly to !L
temperature of 43C., and maintain it On a water bath [It this t.cmpemtnre.
Add 1 ce. of the rennin solution to the milk in one vcssel, and 1 ee. of the
Refcrence Rennin solution to the milk in the other vesscl, noting the time
when (~ach solution is added. Stir each milk mixture slowly for 10 seconds
immediately after the solut.ion is added. Note the time which elapses until
the milk thickens, as shown by a distinctly convex surface when the vessel
is tipped to an angle of about 45."

The rennin solution is added to the milk, and the mixture is

maintained at 430., because rennin exhibits its optimum
activity at that temperature. The mixture i:-; stirred to obtain
an equal distribution of the rennin throughout the mille The
thickening of the milk is caused by the cOIlversion of the soluble
casein in the milk into soluble paracasein and insoluble calcium
paracasein.
3. "The rennin solution should coagulat,e in not less than 90 per cent and
not more than 110 per cent of the time required by the Reference Rennin
solution.
"NOTE: Mille varies in its coagulability. Different lots of milk, or the
same distributor's milk obtained on successive days, will be found to vary in
(loagulation time with the Reference Rennin."
The time required for the coagulation of milk by rennin has
been found to be inversely proportional to the amount of rennin
present. This relationship may be formulated as follows:
eXT = K where C represents the concentration of rennin
and T represents the time during which it acts on the mille and K
is a constant. It is obvious that when the amount of rennin
used is reduced, the time required for the coagulation of a given
amount of milk will be increased.
452 QUANTITATIVE PHARMACEUTICAL CHEMISTRY

The National Formulary specifies that rennin should be

capable of coagulating twenty-five thousand times its weight of
fresh milk. In the assay 0.002 Gm. of l'ennin contained in 1 ce.
is used to coagulate 50 ce. of mille Therefore, 1 Gm. of rennin
is used for each 50/0.002 = 25,000 cc. mille
The relative milk-coagulating power of rennin stronger or
weaker than the specified Oltl'ellgth may be determined by
repeated trials, using larger or smaller quantities of rennin.
Questions and Problems
1. What substances tend to acceleratc the activity of rennin?
2. If a 50 cc. sample of milk is found to have an acidity of 0.175 per cent,
how lUuch normal NaOH solution would be required to reduce .the acidity
to 0.14,5 per cent?
8. Milk usually has a specific gravity of from 1.029 to 1.034. The
National Formulary directs that rennin should be capahle of coagulating
twenty-five thousand times its weight, of fresh mill;:. In the assay, however,
It volume of 50 ce. of milk is employed for each 0.002 Gm. of reimin. If the
milk used has a specific gravity of 1.030, what should he the weight-to-weight,
ratio of the milk coagulated to the rennin?
TABLE LXIX.-OFFICIAL SUBSTANCES AssAYED FOR ENZYME ACTIVITY

Substance Substrate OfIicial requirement

U.S.P.
Malt, extract of. . . . . . . .. Pot.ato sf,arch Converts 5 X its wt. of stluch
into soluble sugars.
Pnncreatin ............. Potato starch Converts 25 X its wt. of starch
into soluble carhohydrates.
Casein COIlverts 2.5 X itH wt. of casein
in to soluble proteoses.
Pepsin. . . . . . . . . . . . . . . .. Egg nlbumen Digests 3,000 to 3,500 X its
wt. of egg !LlbulIlen.
N.F.
Pepsin, compound elixir Egg albumen 100 cc. = 1.75 Gm. of refer-
of ence pepsin.
Pepsin, elixir of. . . . . . . .. Egg !tlbumen 100 ce. = 1.75 Gm. of rell1"-
ence pepsin.
Pepsin, elixir of and reIl- Egg albumen 100 cc. = 2.25 Gm. of refer-
niIl ence pepsin.
Pepsin, glycerite of . . . . .. Egg albumen 100 cc. = 8.75 Gm. of refer-
ence pepsin.
Rennin. . . . . . . . . . . . . . .. Milk Coaguhttcs 25,000 X its wt.
Rennin, elixir of -pepsin Milk FlO cc. = 1:75 GIp.. of 1'efer-
and <mce rennin.
 APPENDIX 453
LOGARITHMS OF NUMBERS*

Proportional parts
Natural
:num.bera
o 2 3 4 5 o 7 8

10 0000004300860128017002120253029403340374 4 8 12172125293337
11 0414 0453 0492 0531 0569 0607 0645 06S2 0719 0755 4 8 1115 19 23 26 30 34
12 0792 0828 OS64 0899 0934 0069 1004 1038 1072 1106 3 7 10 14 17 21 24 28 31
13 1139 1173 1206 1239 12711303 1335 1367 1399 1430 3 6 10 13 16 19 23 26 29
14 14611492 1523 1053 1584 1614 1644 1573 1703 1732 3 6 9 121518212427
15 17611790 18181847187l: 190319311959 1987 2014 3 6 8111417202225
16 2041 20082090 2122 2141: 2175 2201 222722/33 2279 3 5 8 11 13 16 18 21 24
17 2304 2330 2355 2380 2405 2430 2455 2480 2504 2529 2 Ii 7 10 12 15 17 20 22
18 2553257726012625264826722695271827422765 2 13 7 9 12 14 16 1921
19 2788 2810 2833 2856 2878 2900 2923 2945 2967 2989 2 4 7 9 1113 16 18 20
20 301030323054 3075 3096 3118 3139 3160 318132Ql 2 4 0 81113151719
21 322232433263328433043324 3345 336G 338G 3404 2 4 6 81012 14 16 18
22 3424 3444 3404 3483 3502 3522 3541 3560 3579 3508 2 4 6 8 10 12 14 15 17
23 3617 363(J 3655 3G74 3092 3711 3729 3747 3766 3784 2 4 6 7 0 11 13 15 17
24 3802 3820 3838 3856 3874 3892 3909 3927 3945 3962 2 4 5 7 9 1112 14 16
25 3979390740144031404840654082409041164133 2 3 /j 7 0101214 1B
26 4150 4166 41834200 4216 4232 424!J 4265 42131 4298 2 3 5 7 8 10 1113 111
27 4314 4330 4346 4302 4318 4393 4400 4425 4440 4456 2 3 5 6 8 9 11 1314
28 4472 4487 4502 4518 4533 454!) 4li04 4579 4594 4609 2 3 5 6 8 9 11 12 14
29 4624 4639 4654 4669 4683 4698 4713 4728 4742 4757 1 3 4. 6 7 I) 10 12 13
SO 4771 47864800 4814 4829 4843 4857 4871 4886 4900 1 3 4 6 7 9 10 11 13
81 4914 4928 4942 4055 4969 4983 4997 5011 5024 5038 1 3 4 6 7 8'10 11 12
82 5051 5065 5079 5092 5105 5119 5132 5145 5159 5172 1 3 4 li 7 8 9 1112
83 5185 5198 5211 5224 5237 5250 5263 5276 5289 5302 1 3 4 Ii 6 8 9 10 12
84 5315 5328 5340 5853 5866 5378 5391 5403 5416 5428 1 3 4 5 6 8 9 1011
85 5441 5453 5465 5478 5490 5502 5514 5527 5539 5551 1 2 4 .5 6 7 9 1011
86 5563 5575 5587 5509 5011 5623 5635 5647 5658 5670 1 2 4 5 6 7 8 10 11
87 5682569457055717572957405752570357755786 1 2 3 Ii 6 7 8 9 10
88 5798 5800 5821 5832 5843 5855 5866 5877 5888 5899 1 2 3 5 6 7 8 9 1(}
89 5911 5922 5933 5944 5955 5966 5977 5988 5999 6010 1 2 3 4 5 7 8 9 10
.0 6021 6031 604260536064607560811 0096 6107 6117 1 2 3 4 5 6 8 910
41 6128 6138 6140 (1160 (1170 6180 (1101 (l201 6212 6222 1 2 3 4 5 6 7 8 9
42 0232 6243 6253 6263 6274 6284 6294 (1304 (1314. 0325 1 2 3 4 5 6 7 8 9
43 6335 6345 6355 6365 6375 6385 6395 0405 6410 6425 1 2 3 4 5 6 7 8 0
44 64356444645464646474648464936503 G513 0522 1 2 3 4 5 6 7 8 9
45 6532 6542 6551 6561 6571 6580 6590 6599 6609 6618 1 2 3 4 5 6 7 8 9
46 6028 GG37 G640 0656 6065 6675 6684 6693 G702 6712 1 2 3 4 5 6 7 7 8
<\7 6721 6730 6739 6749 6758 6767 6776 6785 6794 6803 1 2 3 4 5 .5 6 7 8
48 . 6812 6821 6830 6839 6848 6857 6866 6875 0884 6893 1 2 3 4 4 5 6 7 8
49 6902 6911 6020 6928 6937 0946 6955 6964 6972 6981 1 2 3 4 4 Ii 6 7 8
110 6990 699870077016 7024 7033 7042 70507059 7067 1 2 3 3 4 5 6 7 8
111 7076 7084170\)3 7101 7110 7118 712G 7135 7143 7152 1 2 3 3 4 5 (3 7 S
52 7160 71GS '1177 7lS5 7193 7202 7210 7218 7226 7231i 1 2 2 3 4 5 6 7 7
53 7243 7251\725!l 7267 7275 7284 7202 7300 7308 7316 1 2 2 3 4 5 6 6 7
54 7324 7332 7340 7348 7356 73134 7372 7380 7388 7396 I 2 2 3 4 5 6 6 7

* See page 13 for exulIlJlles illustrating the llile of Iogurithwio tables in calculations,
454 QUAN'l'I'l'A1'1VE PH~lRM.4_CEUTJC.flL CHEMISTRY

LOGARl'l'HMS OF NUMBERs.-(ConlinHerl)

l'roportion!11 parts
Natural 0 1 2 3 4 5 0 7 8 0

1121314151017181~
numbers

55 7404 7412 7419 7427 7435 7443 7451 7459 7466 747-1 1 2 2 3 <I Ii 5 6 7
56 7482 74DO 7497 71i05 7513 7520 7528 7536 7543 7551 1 2 2 3 <I 5 Ii 6 7
57 7559 7566 71i74 7582 7589 7597 7604 7612 7619 7627 1 2 2 3 4 Ii 5 6 7
58 7634 7642 7649 7657 70134 7672 7679 713SB 7694 7701 1 1 2 3 4 4 5 6 7
59 7709 7716 7723 7731 7738 7745 7752 7700 7767 7774 1 1 2 3 1 4 5 6 7
7782 7825 7832 7839 7846 1 n
60 7789 7796 7803 7810 7818 1 3 <I 4 5 6 6
61 7853 7860 7868 7875 nm~ 7S8!) 7896 7903 7010 791'ij 1 1 2 3 4 4 5 6 6
62 70U 7931 7938 7945 7952 7959 7066 7973 7980 7987 1 1 2 3 3 <I 5 6 6
63 7993 8000 8007 eG14 8021 8028 8035 8041 8048 805& 1 1 2 3 3 'I 5 5 6
64 8062 8069 8075 8082 8089 8006 8102 8109 8116 8122 1 1 2 3 3 'I 5 15 a
65 8129 8136 8142 8149 8156 8162 8169 8176 8182 8189 1 1 2 3 3 'I 5 5 (\
66 8195 8202 8209 8215 3222 8223 0235 8241. 8248 8254 1 1 2 3 3 4 5 5 6
67 8261 8267 8274 8280 8287 8293 8209 830G 8312 8319 1 1 2 3 3 4 5 5 6
68 8325 8331 8338 8341 8351 8357 83G3 8370 8376 8382 1 1 2 3 3 4 4 5 6
69 8388 8395 8401 8407 8414 8420 8426 8432 8439 8445 1. 1 2 .2 3 4 4 5 6
70 8451 8457 8463 8470 8476 8482 8488 8404 8500 8506 1 1 2 2 3 4 4 5 6
71 8513 S5W 8525 8531 8537 3543 35,10 8555 81lGl 8557 1 ,1
1 .2 .2 3 <I 5 5
72 8573 8579 3585 8591 8597 8003 SGOO 8615 8621 8627 1 1 2 2 3 4 4 5 5
73 8633 8639 8641i 8651 8657 86G3 8660 8675 8681 8686 1 1 2 2 3 4 4 5 1>
~\I: 8692 8698 8704 8710 8716 8722 8727 8733 8739 8745 1 1 2 2 3 <I 4 5 5
75 8751 8750 8762 8768 8774 8770 8785 8791 8707 8802 1 1 2 2 3 3 4 5 5
76 8808 8814 [3.820 8825 !l[l31 8837 8842 0843 8854 8859 1 1 2 2 3 3 <I 5 11
77 8805 8871 887G 88S2 3G137 8803 8S!)!) 8904 8910 8015 1 1 2 2 3 3 4 4 5
78 8921 8927 !l932 (lOOS 8943 8949 8954 8960 8965 8971 1 1 2 2 3 3 4 4 5
79 897G 8982 8987 8993 8998 9004 9000 OOlli 9020 9026 1 1 2 2 3 3 4 4 5
80 9031 9036 0042 9047 0053 9058 9063 0069 0074 9079 1 1 2- 2 3 3 4 4 5
81 9085 9090 g0ge 9101 0106 9112 9117 012:3 912[1 9133 1 1 2 2 3 3 4 4 5
82 9138 9143 (l149 9154 0150 9165 9170 9175 (l180 9186 1 1 2 2 3 3 01 4 5
83 9191 9190 9201 9Z0G 0212 9217 9222 9227 9232 9238 1 1 2 2 3 3 1 4 5
84 9243 9248 9253 9258 9263 9269 9274 9279 9281 9289 1 1 2 2 3 3 4 <1 5
85 9204 9299 9304 9a09 1)315 9320 9325 9330 9335 9340 1 1 2 2 3 3 4 4 5
86 9345 9350 9355 9360 D:3Gl; 0370 9375 9380 9385 9390 1 1 2 2 3 3 4 <I II
81 9305 [)400 9405 9410 IH15 9420 9425 9130 0435 0440 0 1 1 2 2 3 3 4 4.
88 11445 9450 9455 94CO 0405 9409 947'1 9479 9484 9480 0 1 1 2 2 3 3 4 4.
89 9494 9499 9504 9509 0513 9518 9523 9528 0533 9538 0 1 1 2 2 3 3 4 <I
90 9542 9547 9552 9557 9562 0566 tl571 9070 9581 9586 0 1 1 2 2 3 3 4 4.
91 9590 9595 0600 0605 9009 9014 0010 9024 9628 0033 0 1 1 2 2 3 3 4 4.
92 9038 9643 9047 O()52 9657 9661 9666 9671 9675 9680 0 1 1 2 2 3 3 4 4
93 9685 9689 9694 0699 9703 9708 9713 9717 0722 9727 0 1 1 2 2 3 3 4 4
94 9731 9736 9741 9745 971)0 9754 9759 9763 9768 9773 0 1 1 2 2 3 3 4 4
95 9777 9782 9786 9791 9795 9800 9805 9809 9814 9818 0 1 1 2 2 3 3 4" 4.
96 9823 9827 9832 9836 9841 9845 9850 980'1 9859 9863 0 1 1 2 2 3 3 4 4
97 9868 9872 9877 9881 9886 9800 98114 9899 9903 9908 0 1 1 2 2 3 3 4 4-
98 9912 9917 9921 9926 9930 9934 0!l39 fJ943 9948 9952 0 1 1 223 3 4 4
99 9956 9961 90U5 9969 9974 9978 9983 9987 9991 9996 0 1 1 223 3 3 4.
 APPENDIX 455
ANTILOGARI'rHMS

I)raportiollal parts
Logarithms 0 1 2 3 .:1 5 0 7 8 0

1\2\3\'\5\0\7\8\9

.00 1000 1002 1005 1007 1009 1012 1014 101611010 1021 O. 0 1 1 1 1 2 2 2
.01 1023 1026 1028 1030 1033 1035 1038 1041) 1042 1045 o 0 1 1 1- I 2 2 2
.02 1047 1050 1052 1054 1057 1059 1002 1064 1067 1009 o 0 1 .1 1 1 2 2 2
.03 1072 1074 1076 1079 10131 1034 1086 1089 1001 1094 a 0 I 1 1 1 2 2 2
.04 1096 1099 1102 1104 1107 1109 1112 1114 1117 1119 a 1 1 1 1 2 2 2 2
.05 1122 1125 1127 1130 1132 1135 1138 1140 1143 1146 0 1 1 1 1 2 2 2 2
.06 1148 1151 1153 1150 115C 1101 1164 1167 1169 1172 0 1 1 1 1 2 2 2 2
.07 1175 1178 1180 1183 1186 1189 1191 1194 1197 1199 0 1 1 1 1 2 2 2 2
.08 1202 1205 1208 1211 1213 1216 1219 1222 1225 1227 0 I 1 1 1 2 2 2 3
.09 1230 1233 1236 1239 1242 1245 1247 1250 1253 1256 0 1 1 1 1 2 2 2 3
.10
.11
1259 1262 1265 1268 1271 1274 1276 1279 1282 128:;
1288 1291 1294 1297 1300 1303 1300 1309 1312 1315
0 1 1 1 1 2 2 2 a
0 1 1 1 2 2 2 2 3
.12 1318 1321 1324 1327 1330 1334 1337 13,10 1313 1340 0 1 1 1 2 2 2 2 3
.13 1349 1352 1355 1358 1361 1365 13G8 1371 1374 1377 0 1 1 1 2 2 2 3 3
.14 1380 1384 1387 1390 1393 1396 1400 1403 1406 gOD 0 1 1 1 2 2 2 3 3
.15 1413 1416 1419 1422 1426 1429 1432 1435 1439 1442 0 1 1 1 2 2 2 3 3
.16 1445 1449 1452 14uo 1450 1402 146G 1409 1472 1470 0 1 1 1 2 2 2 3 3
.17 1479 1483 1486 1489 1493 1490 1500 1503 1507 1510 0 1 1 1 2 ~ 2 a 3
.18 1514 1517 1521 1524 1528 1531 1535 1538 1542 154" 0 1 1 1 2 2 2 3 3
.19 1549 1552 1556 1560 1563 1567 1570' 1574 1578 1581 0 1 1 1 2 2 3 3 a
.20 1585 1589 1592 1596 1600 1603 1607 1611 1614 1618 0 1 1 1 2 2 3 a 3
.21 1622 1626 1620 1033 1637 1641 1044 1648 1052 165G 0 1 1 2 2 2 3 3 3
.22 1660 1663 1667 1671 1675 1679 1683 1687 1690 1094 0 1 1 2 2 2 3 3 3
.23 1698 1702 1706 1710 1714 17W 1722 1726 1730 1734 0 1 1 2 2 2 3 3 4
.24 1738 1742 1746 1750 1754 1758 1762 1760 1770 1774 0 1 1 2 2 2 3 3 4
.25 1778 1782 1780 1791 1791: 1799 1803 1807 18H 1816 0 1 1 2 2 2 3 3 4
.26 1820 1824 1828 18:3!! 18:17 1841 1845 1849 185~ 1858 0 1 1 2 2 3 3 3 4
.27 1862 1866 1871 1875 187e 1884 1888 1892 1897 1901 0 1 1 2 2 3 3 3 4
.28 1905 1910 1914 1919 192~ 1920 1932 1930 1{)41 1945 0 1 1 2 2 3 3 4 4
.29 1950 1954 1950 1903 11)68 1072 1977 1982 1986 1991 0 1 1 2 2 3 3 4 4
.30 1995 2000 2004 2009 2014 2018 2023 2028 2032 2037 0 1 1 2 2 3 3 4 4
.31 2042 2046 2051 n056 Z061 20M ~070 2075 2080 2084 0 1 1 2 2 3 3 4 4
.32 2089 2094 2099 2104 2109 2113 2118 2123 2128 2133 0 2 2 :1 3 4 4
.33 2138 2143 2140 2153 2158 216:1 2168 2173 2178 2183 0 111
1 1 2 2 3 3 4 4
.34 2188 2193 2198 2203 2208 2213 2218 2223 2228 2234 1 1 2 2 3 3 4 4 5
.35 2239 2244 2249 2254 2259 2265 2270 2275 2280 2280 1 1 2 2 3 3 4 4 5
.36 2291 2296 2301 2807 2312 2317 2323 23213 233:) 2330 1 1 2 2 3 3 4 4- 5
.37 2344 2350 2355 2360 2360 2371 2377 2332 238G 2393 1 1 2 2 3 a 4 4 5
.38 2399 2404 2410 2415 2421 2427 2432 243:3 2443 2440 1 1 2 2 3 3 4 4 5
.39 2455 2460 2466 2472 2477 2483 2489 2495 2500 200G 1 1 2 2 3 3 4 5 .5
.40 2512 2518 2523 2529 2535 2541 2547 2553 2559 2561 1 1 2 2 3 4 4 5 5
.41 2570 2576 2582 2588 2594 2600 2606 2612 2618 2024 1 1 2 2 3 4 4 II 5
.42 2630 2636 2642 2649 2655 2661 2667 2673 2679 2685 1 1 2 2 3 4 4 II 6
.43 2692 2698 2704 2710 2716 2723 2729 2735 2742 2748 1 1 2 3 8 4 4 5 6
.44 2754 2761 2767 2773 2780 2786 2793 2799 2805 2812 1 1 2 3 3 4 4 II 0
.45 2818 2825 2831 2838 2844 2851 2858 2864 2871 2877 1 1 2 3 3 4 II II 0
.46 2884 2891 2897 2904 2911 2917 2924 2931 2938 2944 1 1 2 3 3 4 5 II 6
.47 2951 2958 2965 2972 29'19 2985 2992 2999 3006 3013 1 1 2 3 3 4 II 5 6
.48 3020 3027 3034 3041 3048 3055 3062 3069 3076 3083 1 1 2 3 4 4 5 6 6
.49 3090 3097 3105 3112 3119 3126 3133 3141 3148 3155 1 1 2 3 4 4 5 6 6
456 QUANTI1'A'l'IVE PHA.RMACEU'l'ICAL CHEMISTRY

. AN'l'ILOGARiTHlIfs.-(Continued)

Proportion,,! parts
Logarithms Q 1 2 3 4 '5 (\ 7 8 0

1121314\516171819

.50 3162 3170 3177 3184 3192 3199 3200 3214 3221 3228 1 1 2 3 4. 4 5 6 7
.51 3236 3243 3251 3258 3260 3273 3281 3289 3296 3304 1 2 2 3 4 5 Ii 6 7
.52 3311 3319 3327 3334 3342 3350 3357 3365 3373 3381 1 2 2 3 4 5 5 6 7
.53 3388 33HO 3404 3412 3<120 3428 3430 3443 3451 3451) 1 2 2 3 4 Ii 6 B 7
.54 3467 34/15 3483 3491 3499 31i08 3516 352"{ 3532 3540 1 2 2 3 4 Ii 6 a 7

.55 3548 3556 3565 3573 3581 3589 3597 3606 3614 3622 1 2 2 3 4 5 6 7 7
.1i6 3631 3639 3643 3050 3064 3673 3081 3690 31398 3707 1 2 3 3 4 5 6 7 8
.1i7 3715 3724 3733 3741 3750 3768 8767 3776 3784 3703 1 2 3 3 4 5 6 7 8
.58 3802 3811 3819 3823 3837 3840 3855 ~864 3873 3882 1 2 3 4 4 Ii 6 7 8
,.59 3890 3899 3908 3917 3926 3936 3945 3954 3963 3972 1 2 3 4 5 6 6 7 8
.60 3981 3990 3999 4009 4018 4027 4036 4040 4055 4064 1 2 3 4 5 6 6 7 8
.61 1074 4083 4093 4102 4111 4121 4130 4140 4150 4159 1 2 3 4 Ii 6 7 8 9
.62 4169 4178 4188 4198 4::07 4217 4227 4230 42413 4256 1 2 3 4 5 6 7 S 9
.63 42(>6 4270 4285 4295 430t 4315 43.25 4335 4345 4355 1 2 3 4 Ii 6 7 8 9
.64 4305 4375 4,385 4395 4400 4416 4420 4436 4446 4457 1 2 3 4 Ii 6 7 S 9
.65 4407 4477 4487 4498 4508 4510 4520 4539 4550 4560 1 2 3 4 5 0 7 8 9
.66 4571 458! 4592 4003 4(11~ 4024 4034 4045 4650 4607 1 2 3 4 5 6 7 910
.67 4677 4688 4699 4710 4721 4732 1742 47fi3 4761 4775 1 2 3 4 5 7 8 910
.08 4786 4797 4808 4810 4831 4842 4858 J.!:3G4 4875 4887 1 2 3 4 6 7 8 9 10
.69 4898 4909 4920 4932 4114:: 495q 49GO 4977 4989 500e 1 2 3 5 6 7 B 910
.70 5012 5023 li035 5047 fl058 50'70 5082 0093 5105 5117 1 2 4 5 6 7 8 9 11
.71 5129 5140 5152 5104 517C 51133 5200 5212 5224 523G 1 2 4 5 6 7 810 11
.72 5248 5260 5272 5284 5297 m~oo 5321 [j:J:J:J 5310 5358 1 2 4 5 () 7 910 11
.73 5370 5383 5395 5408 542C 54:J~ 5145 5450 5470 5483 1 3 4 5 6 8 910 11
.74 5495 5508 5521 5534 6546 5550 5572 5585 5598 1i(l1O 1 3 4 5 G 8 910 12
.75 5623 563B 5649 5062 S67li 5689 5702 fi715 5728 5741 1 3 4 5 7 8 910 12
.76 5754 5768 5781 57\.)4 58013 5821 5831 li84G 5801 fi875 1 3 4 5 7 8 9 1112
.77 5888 5002 5916 5920 5()43 5057 5970 5084 5998 C012 1 3 ,1 5 7 810 111 2
.78 6020 6039 6053 60G7 6081 6005 61o:l 0124 513:3 G152 1 8 4 (l 7 810 1113
.79 6166 6180 6194 6209 6223 0237 6252 0200 0281 l295 1 3 4 6 7 910 1113
.80 6310 6324 5339 6353 8368 0383 0397 0412 (1427 0442 1 3 4 (l7 910 1213
.81 6457 0471 6480 OUOl 6516 6531 0540 6501 0577 0592 2 3 5 6 8 911 1214
.82 0007 0022 6637 6653 6668 6683 GoOD 0714 (1730 6745 2 3 5 0 8 9 11 1214
.83 6761 6776 6702 6808 6823 6830 68513 6871 3887 0902 2 3 5 6 8 9 1113 14
.84 6918 6931 0950 6966 6982 6998 7015 7031 7047 7003 2 3 5 6 810 1113 15
.85 7079 7090 7112 7129 114e 7161 71713 7194 7211 7228 2 3 /j 7 810 1213 15
.811 7244 7261 7278 7295 7311 732[l 7345 7302 7[l7D 7300 2 3 5 7 810 1213 15
.81 7413 7430 7447 7464 748~ 7409 7510 7531 7M1 700G 2 3 5 7 010 1214 16
.88 7586 7603 7621 7633 705e 7674 7001 7700 7727 7745 2 4 5 7 011 1214 16
.89 7762 7780 7798 7816 7831 7852 7870 7889 71)07 7025 2 4 Ii 7 011 1314 16
.90 7943 7962 7980 7998 8017 8035 8054 8072 8091 8110 2 4 (l 7 911 1315 17
.91 8128 8147 8166 8185 8204 8222 8241 8260 8279 8290 2 1 0 8 911 1315 17
.92 8318 8837 8356 8375 8395 8414 8433 8453 8472 8402 2 4 6 8 10 1214 1517
.93 8511 8581 8551 8570 8590 8610 8630 8650 8670 SOOO 2 4 6 8 10 1214 1618
.94 8710 8780 8750 8770 8790 8810 8831 8851 8872 8892 2 4 0 810 12 1416 18
.95 8913 8933 8954 8974 8995 9016 9036 9057 9078 9099 2 4 6 8 10 12 1517 19
.96 9120 9141 9162 9183 9204 9226 0247 9268 9290 9311 2 4 6 8 11 13 1517 19
.97 9333 9354 9376 9397 9419 9441 9462 9484 0506 9528 2 4 7 9 11 13 1517 20
.98 9550 9572 9594 9616 9638 9661 9683 0705 9727 9750 2 4 7 9 11 13 16 18 20
.99 9772 9795 9817 9840 9863 9886 9908 0931 0951 9977 2 5 7 011 14 1618 20
 INDEX
A Aldehydc content, nssay of voJatile
Abbe refractometer, 242
diagram of, 243
illustration of, 242
temperature cont,rol for, 244
Absolute viscosity, 257
Absorption pipette, 191
Acacia, moisture conteut, deter-
mination of, 331
Accuracy and honesty, 3
Acetylization flask, 368
Acetylsftlicylic add tablets, as,;ayof,
111
Acid number, 345
Acid value, 345
definition of, 345
determination of, 347
tahlc of o"fficial substances with
limits, 348
Acid-base equilibrium, 272
Acidimetry, 105
and alkalimetry, 65
direct titration methods, 105
table of substances assayell by,
112
residual titration methods, 110
table of substances assayed by,
114
Acid-insoluble :18h, 323
Acl(lity index, 345
Adsorbents in alkaloidal assays, 402
Alcohol, determination of, 214
Alcoholic potassium hydroxide, 365
Alcohol-soluble extractive, 339
table of official substances with
limits of, 340
Alcohols, assay of volatile oim for,
368
457
oilll for, 372
Alkalimetry, S()
. direet tikatio!t methorlll, SG
table of oflicial sub~t,utecs as-
sayed by, 91
reHidual titl'l1tiou ll1ethndrs, 90
table of snhHtances lls~ayed hy,
103
Alkaloidal aSHaying, 38G
by aliquot-part method, 403
decanting the aliquot portion,
39(l
detenniuation of the alkaloidal
content, 401
flxtraction of thc drng, 395
shaking out .with add, 3~)S
wit,h immiscible solvc'llt, 399
table of official 8uhsttwces as-
sayed by, 411
emulsification ill, 389
general procedures, 394
type methods, 403
pl'inciples applied in, 3SG
sources of errol' in, 390
by special methods, 418
by total-extractioJl method, 411
percolntor recommended for,
398
table of official sllhstrmces as-
sayed hy, 41G
AlkaJoidal saIt~, ussrlY of, 431
table of those oflicial salts assayed
for content of alka1oid, 43() .
Alkaloidal test solutions, 393
Alkaloic\ttl tit.rations, choice of incli-
cators for, 392
Alkaloids, graviuletric determina-
tion of, 403
volumetric determinntioll of, 407
458 QUAN'1'ITATIVE PHARMACEU'1'ICAL CHEMIS'1'RY
Allyl isothiocyanate, assay for, 382
Aloe, asslty of, 341
Alum, as.'iay of, 50
Aluminum, determination of, 50
Ammonia in water, 261
Ammonium acetate solution, asslty
of,97
Ammonium bromide, assay of, 124
Ammonium hypophosphite, assay
of, 183
Ammon.ium thiocyamtte 0.1 N solu-
tiOIl of, US
Ampere, 305
Amylopsin, 448
Analytical b!1lance, 21
di!1grmn of, 22
figure of, 22
rules for use and care of, 23
sensitivity of, 24
determinn.tion of, 25
zero point cl(iterminatioll, 24
AIl!1lyzer, 248
Angle of incidcnce, 210
of refraction, 210
of 1'0 tutioIl, 248
Anions, 307
Anode, 307
illustratioIlS of, 309
rotating, 311
Antilogarithms, 455
Arecl1, assay of, 409
Arithmeticl1lmeall, ] 0
average d!-1viation from, 10
Aromatic sulphuric acid, assay of,
110
Arsenic tri.oxide, assay of, 162
nephelometric estimation of, 267
Ascaridol content, assny for, 380
Ash content, 323
Ash limits of officinl substances, 327
Atomic weights, table of, inside back
cover
Avogndro's hypothesis, 189
B
Babcock bottle, 384
Balance, analytical, 21
Barium hydroxide, 83
Rtandard solution of, 83
Barium sulfate, determination of
specific gravity of, 220
Belladonna leur, aSRay of, 41(}
extract, assay of, 415
fiuidextrad, nssay of, 416
tincture, assay of, 414
Benzoin, assay of, 339
Block COmp[Lrator, 299
Boiling points, 22G
apparatus for, 234
method of determining, 235
table of official, 238
Borie acid, aRsay of, 107
detel"mination of pH of solution
of, 287
of solubility in water, 205
Boyle's law, 189
Bromere;;ol purple, 73
Bromine, 0.1 N, 180
Bromphenol blue, 73
BromthYIIlol hlue, 73
Buffer mixtures, 294
table of, 294
Buffer solutinns, 293
solutions used to prepare, 293
Buffers, G7
Bunsen valve, 150
Burettes, 58
c!1libration of, G3
illustrations of, 59
official requirements fOl, 59
reading of, 60
C
Caffeine, assay of drugs containing,
429
table of official substances assayed
for, 431
Calcium carbonate, assay of, 146
Calcium gluconate, assay of, 141
Calcium glycerophosphate, assay of,
48
Calculation of results and errors, 9
Calibrat~on of weights, 26
 INDEX 459
Calomel electrode, 279 Copper sulphate. assay of, 172
illustration of, 279 Cottonseed oil, saponification value,
potential of, 280 determinat.ion of, 349
Camphor, determination of specific solidificatioll temperature of fatty
gravity of, 221 acids of, 231
Cantharides, assay of, 437 Coulomb, 30G
Carbon dioxide, assay of, 192 Critical angle, 241
Cathode, 307 Critical my, 241
illustmtiolls of, 310 Croens, assay for 0.0101', 265
Cations, 307 Crucibles, 16
Centipoise, 257 diagrn,m of positioll above flame
Charles' law, 190 20
Chloride ion, determination of, 37 fritt.ed glass, 17
Chlorides, turbidimetric test for, Gooch filtr:ttioll, 16
270 Crude fiber, 336, 342
Chlorinated lime, assay of, 170 Cubic centimeter, 58
Cinchona,. assay of, 405 Cupric sulphate, aSSlLY of, 172
compound tineture, assay of, '106
Citrated caffeine, 431 D
Cleaning mixture, 5
Cluve, crude fiber content, deter- Dalton's )mv, 198
mination of, 342 Decomposition voltage, 308
ether-soluble extmctive content, DeJl~ity, 207
determination of, 338 Desiccators, 21
Cochineal, 74 Deviation, !werage, 10
Coefficient of distribution, 391 Dextrorotatory, 248
Colchicum, assay of, 422 Dichromfltc methods, 151
table of official substances ~Issayed Digitalis leaf, ash eontent, determi-
by the same method as, 425 nation of, 225
Color, assay for, 255 moisture content, determination
Color eomparators, 298 of, 331
Color standards, 297 Diltlted sulfuric acid, assay of, lOt}
Colorimeters, 263 Diphenyll1l11ine indicator, 152
Colorimetric methods, 260 Dil'r.ct titrl1tion, 86
Colorimetry, 250 Distilling points, 234
Common ion effect, 35 apparatus for, 23'1
Comparators, color, 298 method of determining, 235
Compound solution of iodine, assay of volatile oils, 363
of, 164 Distribution constant, 391
Congealing points, 226 Distribution ratio, 391
method of determining, 231 Duhoscq colorimeter, 262, 263
table of officitll, 233
of volatile oils, 363 E
Congo red, 74
Constant weight, 20 Economy of time, 5
Constants of fats, oils, and waxes, Electrical units, 305
345 Electroanalysis, 307
460 QUANTITATIVE PIIARMACEUTICAL CHEMISTRY
Electrochemical equivalent, 306
Electrode, glass, 290
hydl'Ogen, 277
potential, 280
qnillhydrone, 288
Eleetrolysis, 307
Electr(lIyte, 300
Electr(llytic li.pparatus, 309
diagram of connoctions for, 311
Electrolytie methods, 305
aSStLY of coppor sulphate, 313
. asslLY of mercuric chloride, 315
table of substances assnyed by,
319
Electron, 130
Elixir of iron, qUlIllIle and Rtrych-
Hille, pH determination of, 289
Elixir of three bromides, [lasay of,
125
End point, llefinition of, 57
in cliehrollUlte methodH, 152
ill precipitl1tion methods, 11(1
Engler degrees, 259
Enzymes, assay of substlLTloes con-
taining, '144
factors iufluencing activity, 444
Ephedrine slllfllte, assay of, 435
Epinephrine hydrochloride, estima-
tion of colorirnetricaJly, 264
Equivalence point, 65
Errors, indeterminate, 10
nature of, 9
Ester <!ontent, aSSl1Y of volatile oils
for, 364
Ester number, 353
definition of, 353
determination 0[, 353
official substances with required,
353
Et;ter value, 353
Ether-soluble extractive, 337
table of official substances with.
limits of, 338
Ethyl nitrite, assay of spirit of, 178
Eucaine hydrochloride, assay of,. 432
Evaporation of liquids, 19
Extract of beef, estinUttion of
nitrogen in, 100
Extraction, with immiscible sol-
vents, 399
Extractive, alcohol-soluble, 339
ether soluble; 337
purified petroleUIll benzin, 341
water-soluble, 341
Extractive content, 336
F
Flvrndny's laws, 130, 306
Fats, eonstants of, 845
Ferrie !tIntnoniurn sulfnte, indicator
solution, 116
Ferric chloride, lLssay of, Hi8
Ferrous oarbonate, aSSlty of nutss of,
153
Ferrous sulfate, assay of, 136
Fiber, c:rnde, 237
Figures, sigllifimmt, 11
Flasks, rmlibmtion of, 63
Fluidextract, of hellarionm1 lelLves,
determination of alcohol con-
tent of, 2H
of hyoseyamus, lll-iSllY of, 415
Fraetional rlistillntiull of voh.tile oils,
3G3
Frittecl glass crucibles, 17
G
Gas burette, 191
Gas pipctte, 191
Gases, hws uf, 189
Gasometl'ie llwthods, 188
tltble of substances 11sl:\nyed by,
193
Generalopemtions, 16
Glnss crucibles, fritted, 17
Gllts8 electrode, 290
Gracluated eylinc1el's; 62
Gmvimetric analysis, 29
theory of, 29
Gravimetric determinations of al.k;a-.
. Ioids, 403
 INDEX
Gravimetric methods, 37
as~ay of alum, 50
of calcium glycel'OphospIlI1te, 48
of mercuric chloride, 45
of sodiulU chloride, 37
of sodium sulfate, 43
of solution of magnesium
citrate, 52
sample data for, 40.
Grazing incidence, 241
Guarana, 429
H theory of, 70
Hydrastis, assay oi, 403
Hyelrioclic acid, assay of, 124
Hydrochlorie acid, aSS!1Y of, 10.5
norm!11 solution of, 79
potentiometric titration of, 281
HydrocY!1nic acid eontent, assay of
volatile oils for, 378
Hydrogen electrode, 277
illustration of, 278
platinization of, 278
Hydrogen iOIl concentratioIl, 66, 272
determilllltion of, 277
of some official substances, 30.1
Hydrogen ion measurements, colori-
metric methods of, 292
potentiometric methods of, 277
precautions during, 285
scheme of assembly for, 280.
Hydrogen peroxide, assay of solution
of, 139
Hyoscyamus, assay of, 411
extract of, m~say of, 415
fluidextrad of, !tSSllY of, 415
tiricture of, assay, 414
I
Immiscible solvents, 388
extraction with, 399
use in non-alkaloidal assays, 437
Indeterminate errors, 10.
Index of refraction, 240.
Indicator solutions, 73
461
Indicator solutions, sensitivity of, 76
Indicators, 70, 296
for lLlkaloidal titrations, 392
definition of, 70.
diphenylamine, 152
ferric ammoniulll sulfate, 1 Hi
mixed,29(i
potassium chromate, 117
for precipitation llletho,ls, llG
rules for use of, 73
st!U'ch ,ointion, 157
table of cOllllIlonly used, 71
univers:11, 296
used in colorimetric: hydrog(1ll iOIl
mea~uI'CJnents, 29G
Iodine, assay of compound solution
of, 164
nUIIl bel', definition of, 355
official substancl1s with rOCJuire-
ments i'or, 359
0..1 N solution of, 159, HjD
standardizlltion against 0..1 N
sodium thiosulfate, HiO
test solution, 394
titl'f1tioll flask, 358
value, 355
definition of, 355
determination, 357
Iodohromide test solution, 35G
IodoIIletric methods, 155
Ionizatioll, theory of, 29, Gil
Ipecac, assay of, 407
J
Jalap, ass!ty of, 440.
K
Ketone content, assay of volatile
oils for, 375
Kinematic viscosity, 258
determinatioll of, 259
Kjeld!thl apparatus, 99
Kettsdorfer number, 349
Koppescharr's solution, 180
462 QUANTI'l'A'1'IVE PHARMACEU'l'ICAL CHEMISTRY

L N

Laevorotatory, 248 N atUl'e of errors, 9

Laws, Boyle's, 189 Nephelometry, 260, 266
Charles', 190 N esslcr appftratus, 260
Dalton's, 198 Neutralization methods, 65
FaradI1Y's, 98, 300 buffers in, 67
Ohm's, 305 pH value in, 66
Limit test for chlorides and sulfates, theory of, 66
269, 270 N eutri~lization reactions, 69
Liquid petrolatum, determination of Nitrogen estimation, by GUlllling-
kinematic viscosity' of, 259 l{jeldahl method, 98
Liquids, transfer of, 19 with nitrates and nitrites absent,
Liter, nOrIll!11, 58 98
Litmus, 74 with nitmtes present, 99
LoglLl'ithms of numbers, 453 Nitrometer, illuHtration of, 194
use of, 13 preparation ullll teRting of, 195
Lunge nitrometer, 194 NOIl-voh,tile ether-soluble extrac-
tive, 337
M Normal conditiolls, 189
Magnesia magma, assay of, 94 Normal liter, ,12
Magnesium, determination of, 52 Normal solution, 76, 77
Magnesiulll citrate, assay of, 52 Normality, 77
Mass of ferrous carbollate, assay of, Normality factor, 78
153 Notebooks, 4
Materials required, 7, 392 Nux vomiea, aSHaY of, 425
Mayer's reagent, 394
Measuring flasks, 01 o
Melting points, 226
apparatus for taking, 226 Official sample, 8
table of official, 230 OhJll, 305
Meniscus, 60 Oil of bitter almonds, assay for
Mercuric chloride, assay of, 45 henzaldehyde contcnt, 373
Mercuric potassium iodide, 394 assay fOT hydrocyanic [tcid, 379
Mercurous chloride, aSHuy of, 166 of c[traway, assay of, 375
Methenamine, assay of, 95 of chenopodium, I1SSftyof, 380
Methyl orange, 74 of clove, assay of, 377
Methyl red, 75 of mustard, assay of, 382
Milliliter, 58 of orange, determinatioll of refrac-
Moisture content, 321) tive index of, 245
Moisture limits of official Bubstltnces,
of peppermint, assay for total
333
esters, 366
Moisture tube, 332
assay for total menthol, 368
Molal solution, 76
nephelometric estimation of,
Molar solution, 76
preparation of, 185 269
Monochromatic light, 249 Oleoresin of aspidium, assay of, 439
 INDEX
Olive oil, iodine value determination
of, 357 . oils for, 377
Opiuni, assay of, 418
table of official substances assayed
by the same method liS, 422
Optical activity, official substances
with requirements, 254
Optimum temperature of enzymes,
444
Organic solvents, evaporation of, 400
Oxalic acid, 0.1 N solution of, 143
Oxidation, definition of, 130
Oxidation-reduction, dichromate
methods, 151
tahle of subBitLuces assnyed by,
154
direct titration methods, 136
table of substances tlssayed hy,
140
iodomctric methods, 155
standard solutions for, 132
methods, 155
theory of, 129
Oxide, ofiieittl substullces assayed by
ignition to, 51
P assay of, 92
Pancreatin, assay for cnsein diges-
tive power, 4,19
for starch digestive power, 447
Parallax, effect of, 60
Partition coefficient, 391
Pepsin, assay of, 445
Perm!1nganate methods, 133
pH, colorimetrie methods of deter-
mining, 292
indicators and their use, 293
measurements, directions for, 296
meter, 291
potentiometric methods of deter-
mining, 277
relationship to voltage, 283, 284
value, 66
Phenobarbital, assay of tablets of,
441
Phenol, assay of, 181
463
Phenol content, assay of volatile
Phenol red, 75
Phenolpilthulein, 75
Photometric methods of an!lIysis,
260
Physiological methodR, 321
Physiological salt Hoilltioll, determi-
nation of pH of solution of, 287
Pipettes, 60
calibmtioll of, 63
illustrations of, 62
official requirements for, GO
Plane-polurizcdlight, 247
Poise., 257
Polarimeter, 250
diagmm of opt.ical partR of, 250
illnstration of, 251
of Laurent, 251
of Schmidt and IUnBch, 251
Polr\riscope, 251
Polariscope tube, 248
Polarization, definition of, 247
Polarizer, 248
Policemen, (l
Potassiulll and sodium tartrate,
Potassium chlorate, assay of, 149
Potassium chromate, indicator solu-
tion, 117
Potl1ssiulll dinhrolllate, 0.1 N solu-
tion of, 152
Potassium hydroxide, alcoholic,
preparation of, 365
Potassium iodate, 185
~uolar solution of, 185
oflicial substances assayed with,
187
Pot::Lssium iodlde, assay of, 186
test solution, 356
Potassium mercuric iodide test solu-
tion, 394
Potassium permanganate, 0.1 N
solution of, 134
Potential, electrode, 280
Potentiometric methods, 277
464 QUANTITA7'IVE PHARMACEUTICAL CHEMISTRY
Potentiometric titration, of !1cetic
lIeid with sodium hydroxide, 2B7
of hydrochloric acid sodium with
hydroxide, 2Bl
Precipitates, 17
colloidal and fine-grnined, 18
drying and ignition of, 19
filtl':1tion ltml vm~hing of, 17
Pre(~ipit,f1tion methods, 11 ()
by direct titration, 119
table of substanccs assayed by,
121
end point in, 11 ()
indicators for, 117
by residual titmtion, 122
table of substances assayed by,
127
ProlJllbility curve, 10
Proteolytic power, assay of pepsin
for, 445
Proton, 130
Proximate assays, 388
Purified petroleum benzin extractive,
341
Purity and strength requirements, l\
Pycnometer, 208
Geissler type, 210
method of filling, 209
Sprengel-Ostwald, 408
Pyropho!:lphnte, tnble of official sub-
stances assayed as, 54
Q Sedimentation cone, 447
Quinhydronc, 288
Quinhydrone electrode, relation of
pH to voltage, 288
R of, 173
Reactions, in neutralization, ()9
reversible, 30
Reagents, ()
Reduced iron, aSSfW of, 137
Reduction, definition of, 130
Refractive index, 240
Refractive indices, determination of,
245
Refrantive indiees, table of official
suhstances with, 246
of volatile oils, 31i3
Refractometer::;, 241
Relative viscosity, 257
Rennin, aHsay of, 450
ReRidual titration, 57, S()
Residue, official requirements of, 328
Result, rejcetioll of, 11
Results anti errors, 9
Reversible reactions, 30
Riders, 28
Rosin, acid v(tlue, determination of,
347
Rotating anode, device for, 311
Rotatory power, 247
of volatile oils, 3H2
Roulette comparator, 300
s
Snlicylic ncicl, melting point deter-
mination of, 227
SttillpleH, Helection of, 7
drying and ignition of, 19
Sam piing, 7
Saponifientioll value, definition of,
349
determination of, 349
offioial sllb~tances with limits of,
352
S~\ybol(l 1mivel'sll1 ViSCOHity, 259
Separatory funnel, 3B8
Significflnt figures, 11
Silver nitrate, 0,1 N solution of, 117
Sodium ~Lrsenate, exsiccated, [LSsay
Sodium bicarbonate, assay of, 86
Sodium chloride, assay of, 37, 122
Sodium hydroxide, assay of, 87
normal solution of, B1
Sodium nitrite, IIssay of, 144
Sodium salicylate, assay of, 89
tablets, assay of, lOB
Sodium sulfate, assay of, 43
 INDEX
Sodium thiosulfatc, 0.1 N solution,
157
Solidification temperature, deter-
mination of, 231
table of, 233
Solubility, 203
apparatus used to determinc, 204
descriptive terms used ill, 203
factor~ affecting, 204
product, principle, 31
tahle of, 32
of volatile oils, 364
Solution of arsenous acid, nephel-
ometric estimation of arsenic
trioxide in, 267
of epinephrine hydrochloride, de-
termination of pH of, 264
Solutions, classes of, 203
concentrations of standard, 76
of indicators, 73
molal,76
molar, 76
normal,76
Source a.nd natme of errors, 9
Soxhlet extrnction apparatus, 337
Special methods, alkaloidal llssays
by, 418
SpeCific gravity, 207
of fats and oils, 211
official method for, 211
of liquids, 208
methods of determining, 208
method of determining alcohol,
211
of solids, 219
heavier than and insoluble in
water, 219
heavier than and soluble .in
water, 222
lighter than and insoluhle in
water, 220
lighter than and soluble in
water, 223
table of official, 225
of volatile oils, 362
Specific gravity bottle, 208
465
Speeific gravity determinations, by
means of pycnometer, 214
of liquids by means of the West-
plml halance, 216
by the use of hydrollllltl'r:.;, 218
by weighing a solid of known
specific gravity in, 218
Specific rotation, definition of, 249
Specific rotatory power, mLlculation
of,249
determination of, 252
table of official, 2.55
Spirit of ethyl nitrite, assay uf, 179,
196
of peppermint, aHHUY of, 384
nephelometric e~tinmtion of oil
in,2li9
Stalldmd eontiitionR, 189
Standard solution~, 57, 7!i, 132, 1.57
of hydrochloric acicl, 79
in oxidation-reduction methods,
132
of sodiulll hydroxide, 81
of sulfuric acid, 83
0.1 N amllloniulll thiocyanate, 118
0.1 N harillm hydroxide, 83
0.1 N hromine, 180
0.1 N iodine, 159, HiO
0.1 N oxalic acid, 143
0.1 N potassium rliehroIll!tte, 152
0.1 N potassiuIll pcrmallgarmte,
134
0.1 N silver nitrate, 117
0.1 N sodium thio~ulfate, 157
Standardization, 76
Starch digestive power, as~ay of
pancreatin for, 447
indicator solution, 157
test solution, 356
Stoichiometric point, 58, 65
Strong silver-protein, assay of, 120
Success as all analyst, 3
Sucrose, determination of specific
rotation of, 252
Sulfate, determination of, 42
official substances determined as,
44
466 QUAN'l'ITATIVE PHARMACEUTICAL CHEMIS'I'BY
Sulfide, determination of, 45
official substanees determined as,
47
Sulfurie acid, preparation of a 10 per
cent solution of, 219
standard solution of, 83
T for phenol content, 377
Tartaric acid, assa,yof, 110
Test solutions, for alkaloids, 394
Theobrumine with sodium salicylate,
assay of, '132
Theophylline with ~odium acetate,
assay of, 434
Thymol blue, 75
Thyroid, aSSllY of, 175
Titer, 65
Titration, 78
curves, 285
direct, 86
potentioIIll'b-ically, 281'
residual, 86
Titriruetric methods, 57
Toluene moisture method, 331
'l'otal ash, 323
Total esters, assay or oil of pepper-
mint for, 364
Transfer of liquids, 19
Tmhidimetl'ic tests, 2()9
Type process, aliquot, 403
alkaloidal aseays by, 403
general procedure, 394
Type process, total extraction, for
alkaloidal assays, 403
u WllX, yellow, determination of spe-
Units of capacity, 58
of electricity, 305
Unsaponifiable matter, 354
V use of, 217
Viscosimeter, 258
Viscosity, definition of, 256
determination of, 259
units of, 257
Volatile ether-soluble extractive, 337
Volatile oils, assay of, 361
for aleohol content, 3(18
for aldehyde contellt, 372
for ester contcnt, 364
for hydrocyanic acid cont.ent,
378
for ketone content, 375
in spirits, 384
congealing points of, 363
constituents of, 3<l1
dctermination of specific gravity
of,217
distilling points of, 303
fractional distillation of, 363
refmctive index of, 303
rotatory power of, 3112
solubility of, 30'1
spceific gravity of, 362
Volt, 305
Voltagc, relationship to pH, 284
Volullictric analysis, 57
Volumetric apparatus, 58
calibration of, 62
eJeaning of, 62
sources of ei-rur in use of, 64
w
Wagner's reagent, 394
Wash bottles, 5
Water, determination of ammonia
content of, 261
Water-soluble extraetive, 341
table of ofHcirt! substances with
limits of, 341
eifie grnvity of, 222
Weights, calibration of set of, 26
description of, 25
Westpllll1 balnnce, 216
x
Xylene moisture method, 331
z
Zinc oxidb assay of, 92