mupirocin.doc

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CONFIDENTIAL SmithKline Beecham Pharmaceuticals

WorthingWest SussexBN14 8QHT +44 190 8!! 000"""#gs$#com

ANALYTICAL PROCEDURES_SUMMARY

The anal%tical metho&s use& to test 'u(irocin )ree *ci& tos(eciication are the metho&s o the ,uro(ean Pharmaco(oeia-metho&s e.ui/alent to the ,uro(ean Pharmaco(oiea metho&s or inhouse metho&s#

Table 1 Methods Used to Test Mupio!i" Fee A!id to Spe!i#i!atio"

A"al$ti!al Method Method Details

escri(tion escri(tion

&entiication nrare& a2sor(tion s(ectro(hotometr%

'u(irocin content 2% HP3 'u(irocin )ree *ci& ontent 2%HP3

(H Potentiometric &etermination o (H

5elate& Su2stances content 2% HP3 rug 5elate& m(urities 2% HP3

S(eciic 6PT*3 rotation 6(tical rotation

Water content Water semi7micro &etermination

5esi&ual Sol/ents 2% 5esi&ual Sol/ent 2%

SmithKline Beecham Limited

Registered in England and Wales

No. 2337959Registered Ofce9! "reat West Road# Brent$ord

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DESCRIPTION

O##i!ial e#ee"!e i" Peu% Ph& Eu&

*s (er current Ph# ,ur monogra(h or 'u(irocin#




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EUROPEAN P'ARMACOPOEIAL MONO(RAP' FOR INFRARED

A)SORPTION SPECTROP'OTOMETRY

O##i!ial e#ee"!e i" Peu% urrent Ph# ,ur

*&*&*+& A)SORPTION SPECTROP'OTOMETRY, INFRARED

nrare& s(ectro(hotometers are use& or recor&ing s(ectra in theregion o 40007:0 cm;1 or in some cases &o"n to !00cm;1#

APPARATUSS(ectro(hotometers or recor&ing s(ectra consist o a suita2le lightsource- monochromator or intererometer an& &etector#

)ourier transorm s(ectro(hotometers use (ol%chromatic ra&iation an&calculate the s(ectrum in the re.uenc% &omain rom the original &ata2% )ourier transormation#

S(ectro(hotometers itte& "ith an o(tical s%stem ca(a2le o (ro&ucing

monochromatic ra&iation in the measurement region ma% also 2e useNormall% the s(ectrum is gi/en as a unction o transmittance- the.uotient o the intensit% o the transmitte& ra&iation an& the inci&entra&iation# t ma% also 2e gi/en in a2sor2ance#

The a2sor2ance is &eine& as the logarithm to 2ase 10 o thereci(rocal o the transmittance ?




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)& Dis!Triturate 17! mg o the su2stance to 2e examine& "ith 007400 mg-

unless other"ise s(eciie&- o inel% (o"&ere& an& &rie& (otassium2romi&e 5 or (otassium chlori&e 5# These .uantities are usuall%suicient to gi/e a &isc o 1071: mm &iameter an& a s(ectrum osuita2le intensit%# the su2stance is a h%&rochlori&e- it isrecommen&e& to use (otassium chlori&e 5# areull% grin& the mixture-s(rea& it uniorml% in a suita2le &ie- an& su2mit it to a (ressure oa2out 800 'Pa # )or su2stances that are unsta2le un&ernormal atmos(heric con&itions or are h%grosco(ic- the &isc is (resse&in vacuo# Se/eral actors ma% cause the ormation o ault% &iscs- suchas insuicient or excessi/e grin&ing- humi&it% or other im(urities in the

&is(ersion me&ium or an insuicient re&uction o (article siDe# * &isc isreEecte& i /isual examination sho"s lac$ o uniorm trans(arenc% or"hen transmittance at a2out !000 cm;1 in the a2sence o as(eciic a2sor(tion 2an& is less than 0 (ercent "ithout com(ensation-unless other"ise (rescri2e

(ases# ,xamine gases in a cell trans(arent to inrare& ra&iation an&ha/ing an o(tical (ath length o a2out 100 mm#,/acuate the cell an& ill to the &esire& (ressure through a sto(coc$ or

nee&le /al/e using a suita2le gas transer line 2et"een the cell an& thecontainer o the gas to 2e examine

necessar% a&Eust the (ressure in the cell to atmos(heric (ressureusing a gas trans(arent to inrare& ra&iation # To a/oi& a2sor(tion intererences &ue to "ater- car2on&ioxi&e or other atmos(heric gases- (lace in the reerence 2eam- i(ossi2le- an i&entical cell that is either e/acuate& or ille& "ith the gastrans(arent to inrare& ra&iation#

FOR RECORDIN( )Y DIFFUSE REFLECTANCE




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Solids# Triturate a mixture o the su2stance to 2e examine& "ith inel%(o"&ere& an& &rie& (otassium 2romi&e 5 or (otassium chlori&e 5#

Fse a mixture containing a((roximatel% : (er cent o the su2stance-unless other"ise s(eciie rin& the mixture- (lace it in a sam(le cu(an& examine the relectance s(ectrum#The s(ectrum o the sam(le in a2sor2ance mo&e ma% 2e o2taine&ater mathematical treatment o the s(ectra 2% the Ku2el$a7'un$unction#

FOR RECORDIN( )Y ATTENUATED TOTAL REFLECTION*ttenuate& total relection in/ol/es light2eing relecte& internall% 2% a transmitting me&ium- t%(icall% or a

num2er o relections# Ho"e/er- se/eral accessories exist "here onl%one relection occurs#

Pre(are the su2stance as ollo"s# Place the su2stance to 2e examine&in close contact "ith an internal relection element such as&iamon&- germanium- Dinc seleni&e- thallium 2romi&e7thallium io&i&e or another suita2le material o high reracti/e in&ex# ,nsureclose an& uniorm contact 2et"een the su2stance an& the "holecr%stal surace o the internal relection element- either 2% a((l%ing

(ressure or 2% &issol/ing the su2stance in an a((ro(riate sol/ent- thenco/ering the 5, "ith the o2taine& solution an& e/a(orating to&r%ness# ,xamine the attenuate& total relectance s(ectrum#

IDENTIFICACION USIN( REFERENCE SU)STANCES

Pre(are the su2stance to 2e examine& an& the reerence su2stance 2%the same (roce&ure an& recor& the s(ectra 2et"een 40007:0 cm ;1

un&er the same o(erational con&itions# The transmissionminima in the s(ectrum o2taine& "ith the

su2stance to 2e examine& corres(on& in (osition an& relati/e siDe tothose in the s(ectrum o2taine& "ith the reerence su2stance


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When the s(ectra recor&e& in the soli& state sho" &ierences in the

(ositions o the transmission minima - treat thesu2stance to 2e examine& an& the reerence su2stance in the samemanner so that the% cr%stallise or are (ro&uce& in the same orm- or(rocee& as (rescri2e& in the monogra(h- then recor& the s(ectra#




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IDENTIFICATION USIN( REFERENCE SPECTRA

Co"tol o# esolutio" pe#o.a"!e# )or instruments ha/ing amonochromator- recor& the s(ectrum o a (ol%st%rene ilma((roximatel% : =m in thic$ness# The &ierence x 2et"een the (ercentage transmittance at the transmission maximum

* at !8G0 cm;1 an& that at the transmission minimum B at!849#: cm;1 must 2e greater than 18# The &ierence %2et"een the (ercentage transmittance at the transmission maximum at 1:89 cm;1 an& that at the transmission minimum at1:8 cm;1 must 2e greater than 10#

)or )ourier7transorm instruments- use suita2le instrument resolution"ith the a((ro(riate a(o&isation (rescri2e& 2% the manuacturer# Theresolution is chec$e& 2% suita2le means- or exam(le 2% recor&ing thes(ectrum o a (ol%st%rene ilm a((roximatel% : =m in thic$ness# The&ierence 2et"een the a2sor2ances at the a2sor(tion minimum at!8G0 cm;1an& the a2sor(tion maximum at !849#: cm;1is greater than0## The &ierence 2et"een the a2sor2ances at the a2sor(tionminimum at 1:89 cm;1an& the a2sor(tion maximum at 1:8 cm;1 isgreater than 0#08#

#




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Fi/ue *&*&*+&01& T%(ical s(ectrum o (ol%st%rene use& to /eri% theresolution (erormance

Ieriication o the "a/e7num2er scale# The "a/e7num2er scale ma% 2e/eriie& using a (ol%st%rene ilm- "hich has transmission minima at the "a/e num2ers sho"n in Ta2le*&*&*+&01&

Ta2le !#!#!4#71# Transmission minima an& acce(ta2le tolerances o

a (ol%st%rene ilm




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Transmission minima *cce(ta2le tolerance

'onochromatorinstruments

)ourier7transorminstruments

00#0 1#: 1#0

!849#: !#0 1#0

101#! 1#0 1#0

1:8#0 1#0 1#0

11:4#: 1#0 1#0

10!8# 1#0 1#0

Method# Pre(are the su2stance to 2e examine& accor&ing to theinstructions accom(an%ing the reerence s(ectrumAreerencesu2stance# Fsing the o(erating con&itions that "ere use& to o2tain thereerence s(ectrum- "hich "ill usuall% 2e the same as those or

/eri%ing the resolution (erormance- recor& the s(ectrum o thesu2stance to 2e examineThe (ositions an& the relati/e siDes o the 2an&s in the s(ectrum o thesu2stance to 2e examine& an& the reerence s(ectrum are concor&antin the ! s(ectra#

Co.pe"satio" #o ate 2apou a"d at.osphei! !abo" dio3ide#)or )ourier7transorm instruments- s(ectral intererence rom "ater/a(our an& car2on &ioxi&e is com(ensate& using suita2le algorithmsaccor&ing to the manuacturerLs instructions# *lternati/el%- s(ectra can2e ac.uire& using suita2le (urge& instruments or ensuring that sam(le




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an& 2ac$groun& single 2eam s(ectra are ac.uire& un&er exactl% thesame con&itions#

IMPURITIES IN (ASES)or the anal%sis o im(urities- use a cell trans(arent to inrare&ra&iation an& o suita2le o(tical (ath length # )illthe cell as (rescri2e& un&er ases# )or &etection an& .uantiication othe im(urities- (rocee& as (rescri2e& in the monogra(h#

MUPIROCIN FREE ACID CONTENT )Y 'PLC

O##i!ial e#ee"!e i" Peu? n7house metho&

Pepaatio" o# Solutio"sMobille Phase

Mobile PhasePotassium &ih%&rogen (hos(hate 0#0!: ' + &iso&iumh%&rogen (hos(hate 0#0!: '# issol/e #9 g o KH!P64an& #: g o Na!HP64- 2oth (re/iousl% &rie& or ! h at1!0 ! M- in car2on &ioxi&e7ree "ater 5 an& &ilute to1000#0 ml "ith the same sol/ent#Potassium &ih%&rogen (hos(hate 0#008G ' + &iso&ium

h%&rogen (hos(hate 0#00 '# issol/e 1#18 g o KH!P64an& 4#0 g o Na!HP64- 2oth (re/iousl% &rie& or ! h at1!0 ! M- in car2on &ioxi&e7ree "ater 5 an& &ilute to1000#0 ml "ith the same sol/ent#iso&ium tetra2orate 0#01 '# issol/e #80 g oNa!B46G-10H!6 in car2on &ioxi&e7ree "ater 5 an& &iluteto 1000#0 ml "ith the same sol/ent# Store (rotecte& romatmos(heric car2on &ioxi&e#So&ium car2onate 0#0!: ' + so&ium h%&rogen car2onate0#0!: '# issol/e !#4 g o Na!6 an& !#09 g o NaH6in car2on &ioxi&e7ree "ater 5 an& &ilute to 1000#0 ml "ith

the same sol/ent# Store (rotecte& rom atmos(heric car2on&ioxi&e#alcium h%&roxi&e- saturate& at !: M# Sha$e an excess o




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calcium h%&roxi&e 5 "ith car2on &ioxi&e7ree "ater 5 an&&ecant at !: M# Store (rotecte& rom atmos(heric car2on&ioxi&e#

ST65*,Store 2uer solutions in suita2le chemicall% resistant- tightcontainers- such as t%(e glass 2ottles or (lastic containerssuita2le or a.ueous solutionsPepaatio" o# Sa.ple Solutio"

*ccuratel% "eigh 4:7:: mg 'u(irocin )ree *ci& into a !00 m3 /olumetric as$#issol/e the material in : m3 o methanol# ,nsure com(lete &issolution- usingsonication i necessar%# ilute to /olume using 0#1 ' ammonium acetate solution(H :#G an& mix thoroughl%#

Pepaatio" o# Sta"dad Solutio"

*ccuratel% "eigh 4:7:: mg 'u(irocin )ree *ci& into a!00 m3 /olumetric as$# issol/e the material in : m3 o methanol# ,nsurecom(lete &issolution- using sonication i necessar%# ilute to /olume using 0#1 'ammonium acetate solution (H :#G an& mix thoroughl%#

Cho.ato/aphi! Paa.etes

olumn? *((rox !: cm containing asuita2le 2on&e& 8 silica (ac$ing

'o2ile (hase? 0#1 ' ammonium acetate ? TH)

)lo" rate? 1#0 m3Amin

etection? FI at !0 nm

*ttenuation? *2out 0#: *F)S

nEection Iolume? 10 =3

Po!edueSet u( the HP3 s%stem using the instrument (arameters &escri2e&

an& ensure the s%stem suita2ilit% criteria are met# nEect 10 =3 ali.uotso sam(le an& stan&ar& solutions an& measure the (ea$ areas# The




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retention time o 'u(irocin an& &rug7relate& im(urities are gi/en inTa2le !

Table *& T$pi!al Rete"tio" Ti.es

Co.po"e"t Mi"utes

ea& /olume 1#0

B53 :G98 #8

B53 !!89 #G

'u(irocin 8#

Cal!ulatio"

'u(irocin content @ Sam(le area x Stan&ar& Wt x St& Purit%Stan&ar& *rea x Sam(le Wt

S$ste. Suitabilit$

)rom six re(licate inEections o the stan&ar&- calculate the relati/e

stan&ar& &e/iation o the (ea$ area#

RSD@ Stan&ar& &e/iation o the (ea$ area x 100

*ritmetic mean o the (ea$ area

The 5S shoul& not 2e greater than !#

alculate the resolution actor 2et"een (seu&omonic aci&

an& 'u(irocin- the tailing actor o 'u(irocin an&the num2er o theoretical (lates using a solution o the stan&ar&


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5esolution actor @ !

W!+ W1Where

T1- T! @ 5etention time o the ! (ea$s

W1- W! @ Basal (ea$ "i&ths o the ! (ea$s

The resolution actor shoul& not 2e less than #:#




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Tailing actor @ W 0#0:

!)

Where?

W4&45 @ Pea$ "i&th o 'u(irocin at : (ea$ height#) @ Wi&th along lea&ing e&ge o (ea$ to (oint

2eneath (ea$ a(ex along : (ea$ height time#

The tailing actor shoul& not 2e greater than 1#:#

Num2er o theoretical (lates @ :#:4: 1 !W0#:

Where?

T @ 5etention time o 'u(irocinW0#: @ Pea$ "i&th o 'u(irocin at hal (ea$ height#

Num2er o theoretical (lates shoul& not 2e less than 4000#




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EUROPEAN P'ARMACOPOEIAL MONO(RAP' FOR

POTENTIOMETRIC DETERMINATION OF P'

O##i!ial e#ee"!e i" Peu? urrent Ph# ,ur

*&*&6 POTENTIOMETRIC DETERMINATION OF P'

The (H is a num2er "hich re(resents con/entionall% the h%&rogenion concentration o an a.ueous solution# )or (ractical (ur(oses- its&einition is an ex(erimental one# The (H o a solution to 2eexamine& is relate& to that o a reerence solution 2% the

ollo"ing e.uation?

(h @ (H J , 7 ,: $

in "hich , is the (otential- ex(resse& in /olts- o the cell containingthe solution to 2e examine& an& Es is the (otential- ex(resse& in/olts- o the cell containing the solution o $no"n (H - $ is thechange in (otential (er unit change in (H ex(resse& in /olts- an&

calculate& rom the Nernst e.uation#

Table *&*&6&01& 7 8alues o# 9 at di##ee"t te.peatues

Te.peatue :;C< 9 :8C# Sha$e anexcess o +H:K6/igorousl% "ith !abo" dio3ide0#ee ate Rat!: M# )ilter or &ecant# Pre(are imme&iatel% 2eore use#




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Potassiu. dih$do/e" phosphate 4&4*5 M ? disodiu.

h$do/e" phosphate 4&4*5 M# issol/e #9 g o KH!P64 an: g o Na!HP64- 2oth (re/iousl% &rie& or ! h at 1!0 ! M- incar2on &ioxi&e7ree "ater 5 an& &ilute to 1000#0 ml "ith the samesol/ent#

Potassiu. dih$do/e" phosphate 4&44=@ M ? disodiu.h$do/e" phosphate 4&4646 M# issol/e 1#18 g o KH!P64 an&4#0 g o Na!HP64- 2oth (re/iousl% &rie& or ! h at 1!0 ! M- incar2on &ioxi&e7ree "ater 5 an& &ilute to 1000#0 ml "ith the samesol/ent#

Disodiu. tetaboate 4&41 M# issol/e #80 g o Na!B46G-10H!6 in car2on &ioxi&e7ree "ater 5 an& &ilute to 1000#0 ml "iththe same sol/ent# Store (rotecte& rom atmos(heric car2on &ioxi&e#

Sodiu. !abo"ate 4&4*5 M ? sodiu. h$do/e" !abo"ate 4&4*5M# issol/e !#4 g o Na!6 an& !#09 g o NaH6 in car2on&ioxi&e7ree "ater 5 an& &ilute to 1000#0 ml "ith the same sol/ent#Store (rotecte& rom atmos(heric car2on &ioxi&e#

Cal!iu. h$do3ide, satuated at *5 >C# Sha$e an excess ocalcium h%&roxi&e 5 "ith car2on &ioxi&e7ree "ater 5 an& &ecant at!: M# Store (rotecte& rom atmos(heric car2on &ioxi&e#

STORA(EStore 2uer solutions in suita2le chemicall% resistant- tightcontainers- such as t%(e glass 2ottles or (lastic containers suita2leor a.ueous solutions




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DRU( RELATED IMPURITIES )Y 'PLC

Pepaatio" o# Solutio"s

Pepaatio" o# Mobile PhasePre(are a 0#1 ' ammonium acetate solution an& a&Eust to (H :#G"ith small amounts o acetic aci 'ix the 2uer "ithtetrah%&rouran in the ratio G0 2uer? 0 tetrah%&rouran# )ilteran& &egas 2eore use#

Pepaatio" o# Dilue"t

Pre(are a 0#1 ' so&ium acetate solution an& a&Eust to (H 4#0 "ithsmall amounts o acetic aci 'ix the 2uer "ith methanol in theratio #

Pepaatio" o# Sa.ple Solutio"*ccuratel% "eigh a2out 0#1 g o sam(le into a 10m3 las$# issol/ein an& &ilute to /olume "ith &iluent#

Pepaatio" o# Sta"dad Solutio"

*ccuratel% "eight a2out 0#0: g o the la2orator% reerence stan&ar&into a !00 m3 /olumetric las$# issol/e in an& &ilute to /olume "ith&iluent#

Cho.ato/aphi! Paa.etes

olumn? *((rox !: cm containing asuita2le 2on&e& 8 silica (ac$ing

'o2ile (hase? 0#1 ' ammonium acetate ? TH)

)lo" rate? 1#0 m3Amin




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etection? FI at !40 nm

*ttenuation? 0#0! *F)S

nEection Iolume? 10 =3

Po!edue

Set u( the HP3 s%stem using the instrument (arameters &escri2e&an& ensure the s%stem suita2ilit% criteria are met# nEect six ali.uotso the stan&ar& solution ollo"e& 2% an ali.uot o eachsam(le#

'easure the (ea$ areas o all com(onents- exclu&ing those &ue to&iluent eects- in the sam(le solutions# 'easure the (ea$ areas o'u(irocin in the stan&ar& solutions# T%(ical retention times aregi/en in Ta2le #

Table 6& T$pi!al Rete"tio" Ti.es

om(onent 'inutes 5elati/e to 'u(irocin

B53 :G98 B53 4:!94 B53 4:!9: B53 !!89 B53 !0!G 'u(irocin B53 4:!9B53 4:!9GB53 4:!98B53 !0!8G

:G#:89101!141:!:!G

0#4!0#0#G0#G:0#871#1G1#!:!#08!#!:

Cal!ulatio"




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n&i/i&ual im(urit% @ * x St& Wt x S(l Iol x St& Purit%

*1 x St& Iol x S(l Wt

Total im(urities @ * x St& Wt x S(l Iol x St& Purit%*1 x St& Iol x S(l Wt

Where?

* @ *rea o an in&i/i&ual im(urit% in the sam(le

*1@ is the area o mu(irocin in the stan&ar&

* is the sum o the areas o the in&i/i&ual im(urities in thesam(le#

S$ste. Suitabilit$

)rom at least six inEections o the stan&ar& calculate the relati/e

stan&ar& &e/iations o the (ea$ area o 'u(irocin#

5S @ Stan&ar& &e/iation o the (ea$ area x 100 *rithmetic mean o the (ea$ area

The 5S shoul& not 2e greater than :#

alculate a resolution actor 2et"een (seu&omonic aci& an& 'u(irocin- the tailing o 'u(irocin an& the num2er otheoretical (lates using a solution o the stan&ar


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5esolution actor @ ! W!+ W1

Where?

T1- T! @ 5etention time o the ! (ea$s

W1- W! @ Basal (ea$ "i&ths o the ! (ea$sThe resolution actor shoul& not 2e less than #:#

Tailing actor @ W0#0: !)

Where?

W0#0: @ Pea$ "i&th o 'u(irocin at : (ea$ height

) @ Wi&th along lea&ing e&ge o (ea$ to (oint 2eneath(ea$ a(ex along : (ea$ height line#

Num2er o theoretical (lates @ :#:4: 1 !W0#:

Where?

T @ retention time o 'u(irocin

W0#: @ (ea$ "i&th o 'u(irocin at hal (ea$ height#

The tailing actor shoul& not 2e greater than !#0#




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EUROPEAN P'ARMACOPEIAL MONO(RAP' FOR OPTICAL

ROTATION

O##i!ial e#ee"!e i" Peu? urrent Ph# ,ur

6(tical rotation is the (ro(ert% &is(la%e& 2% chiral su2stances orotating the (lane o (olarisation o (olarise& light#6(tical rotation is consi&ere& to 2e (ositi/e or &extrorotator%su2stances


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The s(eciic o(tical rotation Rm!0o a su2stance in solution is theangle o rotation - ex(resse& in &egrees - o the (lane o

(olarisation at the "a/elength o the 7line o so&ium measure& at !0 M in a solution o the su2stance to 2e examine& an&calculate& "ith reerence to a la%er o 1 &m containing 1 gAml o thesu2stance# The s(eciic o(tical rotation o a su2stance in solution isal"a%s ex(resse& "ith reerence to a gi/en sol/ent an& concentration#

n the con/entional s%stem a&o(te& 2% the Pharmaco(oeia the s(eciico(tical rotation is ex(resse& 2% its /alue "ithout units the actual units-&egree millilitres (er &ecimetre gram RCmlC&m;1Cg;1 are un&erstoo

The con/ersion actor rom the nternational S%stem to thePharmaco(oeia s%stem is the ollo"ing?

Rmt @ Rt x 0#1G4:

n certain cases s(eciie& in the monogra(h the angle o rotation ma%2e measure& at tem(eratures other than !0 M an& at other"a/elengths#

The (olarimeter must 2e ca(a2le o gi/ing rea&ings to the nearest0#01M# The scale is usuall% chec$e& 2% means o certiie& .uartD (lates#The linearit% o the scale ma% 2e chec$e& 2% means o sucrosesolutions#

Method# etermine the Dero o the (olarimeter an& the angle orotation o (olarise& light at the "a/elength o the 7line o so&ium at !0 0#: M# 'easurements ma% 2e carrie& out at othertem(eratures onl% "here the monogra(h in&icates the tem(erature

correction to 2e ma&e to the measure& o(tical rotation# etermine theDero o the a((aratus "ith the tu2e close& or li.ui&s the Dero is




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&etermine& "ith the tu2e em(t% an& or soli&s ille& "ith the (rescri2e&sol/ent#

alculate the s(eciic o(tical rotation using the ollo"ing ormulae#)or neat li.ui&s?

Rm!0 @ VVl!0

)or su2stances in solution?

Rm!0 @ 1000 l c

"here c is the concentration o the solution in grams (er litrealculate the content c in grams (er litre or the content cin (er centmAm o a &issol/e& su2stance using the ollo"ing ormulae?

@ angle o rotation in &egrees rea& at !0 0#:M-l @ length in &ecimetres o the (olarimeter tu2e-X!0 @ &ensit% at !0 M in grams (er cu2ic centimetre#

)or the (ur(oses o the Pharmaco(oeia- &ensit% is re(lace& 2%relati/e &ensit% -




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*&5&1* ATER% SEMI0MICRO DETERMINATION

The semi7micro &etermination o Water is 2ase& u(on the .uantitati/ereaction o "ater "ith sulur &ioxi&e an& io&ine in a suita2le anh%&rousme&ium in the (resence o a 2ase "ith suYcient 2uering ca(acit%#

AppaatusThe a((aratus consists o a titration /essel "ith? ! i&entical (latinum electro&es tight inlets or intro&uction o sol/ent an& titrant an inlet or intro&uction o air /ia a &esiccant

a sam(le inlet Ytte& "ith a sto((er or- or li.ui&s- a se(tum#nlet s%stems or intro&uction o &r% nitrogen or or as(iration osol/ents ma% also 2e Ytte

The titration is carrie& out accor&ing to the instrument su((lierLsinstructions# are is ta$en throughout the &etermination to a/oi&ex(osure o reagents an& sol/ents to atmos(heric moisture# The en&7(oint is &etermine& using ! i&entical in&icator electro&es comrecte& toan electrical source that maintains 2et"een the electro&es either a

constant current or a constant /oltage# Where &irect titration is use& - a&&ition o titrant causes either a &ecrease in /oltage "hereconstant current is maintaine& or an increase in current "here constant/oltage is maintaine&- until the en&7(oint is reache nstrmnents "ithautomatic en&7(oint &etection are commonl% use

Stan&ar&isation# To the titration /essel- a&& methanol 5- &rie& inecessar%- or the sol/ent recommen&e& 2% the su((lier o the titrant#Where a((lica2le or the a((aratus use&- eliminate resi&ual "ater

Yrom the measurement cell or carr% out a (re7titration# ntro&uce asuita2le amount o "ater in an a((ro(riate orm


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reerence material> an& carr% out the titration- stirring or the necessar%time# The "ater e.ui/alent is not less than 80 (er cent o that in&icate&

2% the su((lier# Stan&ar&ise the titrant 2eore the Yrst use an& atsuita2le inter/als thereater#Fnless other"ise (rescri2e&- use 'etho& *#

Method A# ntro&uce into the titration /essel methanol 5- or the sol/entin&icate& in the monogra(h or recommen&e& 2% the su((lier o thetitrant# Where a((lica2le or the a((aratus use&- eliminate resi&ual"ater rom the measurement cell or carr% out a (re7titration# ntro&ucethe su2stance to 2e examine& ra(i&l% an& carr% out the titration- stirringor the necessar% extraction time#

Method )# ntro&uce into the titration /essel methanol 5- or the sol/entin&icate& in the monogra(h or recommen&e& 2% the su((lier o thetitrant# Where a((lica2le or the a((aratus use&- eliminate resi&ual"ater rom the measurement cell or carr% out a (re7titration# ntro&ucethe su2stance to 2e examine& ra(i&l% an& in a suita2le state o&i/ision# *&& an accuratel% measure& /olume o the titrant- suYcient togi/e an excess o a2out 1 m3 or the (rescri2e& /olume# *llo" to stan&(rotecte& rom light or 1 min or the (rescri2e& time- "ith stirring#

Titrate the excess o reagent using methanol Ror the (rescri2e&sol/ent- containing an accuratel% $no"n .uantit% o "ater#Suitabilit$The accurac% o the &etermination "ith the chosen titrantmust 2e /eriie& or each su2stance to 2e examine The ollo"ing(roce&ure- gi/en as an exam(le- is suita2le or sam(les containing !#:7!: mg o "ater#

The "ater content o the su2stance to 2e examine& is &etermine&using the reagentAsol/ent s%stem chosen# Thereater- se.uential $no"namounts o water Rare a&&e& in an a((ro(riate orm an& the cumulati/e "ater content &etermine& ater each




%iddlese 'W 9"3
http://www.uspbpep.com/bp2008/data/2542.asphttp://www.uspbpep.com/bp2008/data/2542.asphttp://www.uspbpep.com/bp2008/data/1380.asphttp://www.uspbpep.com/bp2008/data/1380.asphttp://www.uspbpep.com/bp2008/data/2542.asp


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a&&ition# alculate the (ercentage reco/er% at each (oint using theollo"ing ex(ression?

W1@ amount o "ater a&&e&- in milligrams

W!@ amount o "ater oun&- in milligrams#

alculate the regression line o the cumulati/e "ater &etermine&against the "ater a&&e alculate the slo(e - the interce(t "iththe y7axis an& the interce(t o the extra(olate& cali2ration line "ith

the x7axis #

alculate the (ercentage mean reco/er% # alculate the (ercentageerrors using the ollo"ing ex(ressions?

a@

the y7axis interce(t- in milligrams o "ater

d@

thex7axis interce(t- in milligrams o "ater

M@ "ater content o the su2stance- in milligrams o "ater#

The reagentAsol/ent s%stem is consi&ere& to 2e acce(ta2le i? Ze1Z an& Ze!Z are not greater than !#: (er cent bis 2et"een 0#9G: an& 1#0!:




%iddlese 'W 9"3
http://www.uspbpep.com/bp2008/bp2008/bp_images/bp2008_v4_01_10_09_appendix_ix/large/2512water_2_2008_57_eq.pnghttp://www.uspbpep.com/bp2008/bp2008/bp_images/bp2008_v4_01_10_09_appendix_ix/large/2512water_1_2008_57_eq.png


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RESIDUAL SOL8ENTS )Y (CO##i!ial e#ee"!e i" Peu% I"0house .ethod

Sa.ple a"d Sta"dad Pepaatio"

Sta"dad Pepaatio"

To a :0 m3 /olumetric las$ a&& a2out 40 m3 o !7methox%ethanol# *&&!: =3 o each o the ollo"ing sol/ents?

He(tane'eth%l iso2ut%l $etone

,th%l acetate*cetone'a$e to /olume "ith ! methox%ethanol#

Sa.ple Pepaatio"

Weigh a2out 1g o sam(le into a 10m3 /olumetric las$- &issol/e an&&ilute to /olume "ith ! methox%ethanol#

Po!edue



No. 2337959Registered Ofce

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