Pharmaceutical Development

Training Workshop on Pharmaceutical Development with focus on Paediatric Formulations

Protea Hotel Victoria Junction, Waterfront Cape Town,
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outh !frica

Date" #$ to %& !pril %&&'

Slide 1 of 43 April 2007

Pharmaceutical Development
Analytical Method Development

Presenter:

Jnos Pogny, pharmacist, PhD pogany.janos@chello.hu WHO expert

Slide 2 of 43

April 2007

Analytical

etho! De"elopment

Outline and Objectives of presentation

#ntro!uction, gui!elines Dossier re$uirements
% Assay % &elate! su'stances % Other issues

ain points again

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April 2007

Training Workshop on Pharmaceutical Development with focus on Paediatric ormulations

#ntro!uction, gui!elines

!nterchangeability "!#$
#()*&+HA(,*A-#.#)/ 0#+1 #+ O2 3.)#4O3&+* 2PPs 5 0*44*()#A. 4# #.A&#)/ W#)H #((O6A)O& 2PP1 2PP 5

PHA& A+*3)#+A. *73#6A.*(+* 0P*1 P* 8 -#O*73#6A.*(+* 0-*1 #+ 5 P* 8 -*

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April 2007

Pharmaceutical e%uivalence
2PPs meet same or compara'le stan!ar!s 0e.g., mar9eting authori:ation, analytical metho!s1 metho!s
% % % % 4ame AP# 0chemical an! physical e$ui"alence1 4ame !osage ;orm an! route o; a!ministration 4ame strength +ompara'le la'eling

Pharmaceutical !e"elopment e$ui"alence 4ta'ility e$ui"alence WHO<, P 0manu;acturing e$ui"alence1

Slide 6 of 43 April 2007

Pre%ualification re%uirements
Analytical metho! "ali!ation is re$uire! 'y WHO ;or the pre$uali;ication o; pro!uct !ossiers. (on<compen!ial A&6 AP#s an! 2PPs =ere>are teste! =ith metho!s !e"elope! 'y the manu;acturer. Analytical metho!s shoul! 'e use! =ithin , P an! ,.P en"ironments, an! must 'e !e"elope! using the protocols an! acceptance criteria set out in the #+H gui!elines 7?0&@1

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April 2007

&uidelines used in P'P

AWHO<, P B.@@ A#t is o; critical importance that particular attention is pai! to the "ali!ation o; analytical test metho!s, automate! systems an! cleaning proce!ures.C Appen!ix B. Analytical metho! "ali!ation 0in WHO Expert
Committee on Specifications for Pharmaceutical Preparations. 40th Report. ,ene"a, WHO, ?DDE 0WHO )echnical &eport 4eries, (o. FGH1. http:>>=h$li'!oc.=ho.int>trs>WHOI)&4IFGHIeng.p!;

6ali!ation o; analytical proce!ures: text an! metho!ology 7?0&@1 #+H Harmoni:e! )ripartite ,ui!elines, 0?DDJ1
http:>>===.ich.org>.O->me!ia> *D#AB@H.p!;

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April 2007

&eneral re%uirements
7uali;ie! an! cali'rate! instruments Documente! metho!s &elia'le re;erence stan!ar!s 7uali;ie! analysts 4ample selection an! integrity +hange control
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Slide 9 of 43

April 2007

Measure of variation "spread of data$

68.26%

95.46%

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April 2007

Mean "average$ chart

Abnormal variation of process special causes USL Upper specification limit

avera e ! mean

Normal variation due to common causes

LSL

Lower specification limit

Abnormal variation of process special causes

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April 2007

#apable process
Almost all the measurements o; a sta'le process ;all insi!e the speci;ication limits USL LSL Cp E K @D @?

@.DD @.GG @.EE ?.DD E ppm ? pp'

Oo4 results: .?HL.

EB ppm

http())www*itl*nist*gov)div+,+)handbook)pmc)section-)pmc-.*htm
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April 2007

/01!2AP!/0 3 2eference 4tandard

Injection 1 2 3 4 5 6 System s ita!i"ity (#e$ i#ement RSD is %MT 0.85%
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tR 6.160 6.167 6.166 6.172 6.165 6.168 6.166 0.004 0.06%

A 12466 12311 12432 12530 12457 12479 12446 74 0.59%

Mean STD RSD

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April 2007

Training Workshop on Pharmaceutical Development with focus on Paediatric ormulations

Dossier re$uirements

5se of analytical methods 6 generics

&"inica" Met(o)s At initia" *(ase o+ *(a#mace tica" )e,e"o*ment
"o develop a stable and reproducible formulation for t$e manufacture of bioe%uivalence& dissolution& stabilit# and pilot' scale validation batc$es ' "o understand t$e profile of related substances and to stud# stabilit# and start measurin t$e impact of (e# product and manufacturin process parameters on consistent )** %ualit#

'(a#mace tica"

"o determine bioavailabilit# in $ealt$# volunteers

At a),ance) *(ase o+ *(a#mace tica" )e,e"o*ment

"o prove bioe%uivalence after critical variations to t$e pre%ualified dossier "o optimi+e& scale'up& and transfer a stable and controlled manufacturin process for t$e pre%ualification product "o be robust& transferable& accurate& and precise for specification settin & stabilit# assessment& and ,- release of pre%ualified batc$es

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April 2007

Analytical procedure characteristics

)ype o; characteristic Accuracy Precision 4peci;icity Detection limit 7uantitation limit .inearity &ange &o'ustness
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#!enti;ication < < 8 < 6 < < 8

April 2007

#mpurities
7uantitati"e .imit

Assay < < 8 8 < < < 8 8 8 8 < < 8 8 8

8 8 8 < 8 8 8 8

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Accuracy 6 !4O 7897 -6.

Accuracy

Trueness

Precision
$2andom errors"

4ystematic errors

!ntra6assay variability

!ntra6laboratory variability

!nter6laboratory variability

2epeatability
4ource: #4O. @FFB. #4O JH?J @<E: Accuracy 0)rueness an! Precision1 o;

!ntermediate precison
easurement

2eproducibility

etho!s an! &esults. #4O, ,ene"a, 4=it:erlan!.

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April 2007

Accuracy and precision

.naccurate and imprecise

!naccurate : imprecise

!naccurate but precise

*recise

Accurate
April 2007

Accurate but imprecise

Accurate and precise

Slide 18 of 43

Percent accuracy "hypothetical figures$

Nevirapine& m Sample 0 3 4 4 5 6 & %LA 51 81 91 011 031 041 Added 1.499 1.814 1.896 0.110 0.300 0.399 6ecovered 1.495 1.810 1.895 0.115 0.319 0.396 6ecover# % 99.3 99.9 99.9 011.4 99.9 99.9 6S7 % 1.64 1.40 1.38 0.99 1.49 0.03

:ean 1.909 1.908 99.9 1.88 "$e data s$ow t$at t$e recover# of anal#te in spi(ed samples met t$e evaluation criterion for accurac# /011 2 3.1% across 51041% of tar et concentrations5.
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April 2007

Percent accuracy "hypothetical figures$

014 014 013 010 011 99 99 98 96 1 0 3 4 4 5 6 8 % m!e# o+ sam*"es

% Reco,e#y

2ed line( ;A
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&reen lines( 54; and ;4;

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April 2007

Precision "of any process$

Meas #e) mean Rea" mean

)he precision 06A&#A-#.#)/1 o; an analytical proce!ure is usually expresse! as the stan!ar! !e"iation 041, "ariance 04?1, or coe;;icient o; "ariation 05 relati"e stan!ar! !e"iation, &4DL.1 o; a series o; measurements. )he con;i!ence inter"al 0+#1 shoul! 'e reporte! ;or each type o; precision in"estigate!.

'R-&ISI.%
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April 2007

2epeatability "of any process$

Meas #e) mean

&epeata'ility expresses the precision 0sprea! o; the !ata, "aria'ility1 un!er the same operating con!itions o"er a short inter"al o; time. &epeata'ility is also terme! intra<assay precision.

R-'-ATA/I0IT1
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April 2007

2epeatability "hypothetical figures$

Injection 1 2 3 4 5 6 Mean STD RSD &I 95%
Slide 23 of 43

'ea2 a#ea 57935 57833 57497 57617 57778 57231 57649 257 0.4% 270
April 2007

Im*1 0.301 0.301 0.299 0.300 0.301 0.298 0.300 0.0013 0.4% 0.0014

)he repeata'ility precision o'taine! 'y one analyst in one la'oratory =as @.?JL &4D ;or the analyte an!, there;ore, meets the e"aluation criterion o; &4D M?L.

!ntermediate Precision and 2eproducibility "of any process$

Meas #e) means

#nterme!iate precision expresses =ithin<la'oratories "ariations. N@, N? an! NG: !i;;erent !ays, !i;;erent analysts, !i;;erent 0manu;acturing1 e$uipment, etc. &epro!uci'ility expresses the precision 'et=een la'oratories N@, N? an! NG 0colla'orati"e stu!ies, usually applie! to stan!ar!i:ation o; metho!ology1. 0)rans;er o; technology1

Inte#me)iate *#ecision o# Re*#o) ci!i"ity

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April 2007

!ntermediate precision "ruggedness$

Sample 0 3 4 4 5 6 :ean S"7 6S7 -. 95%
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5Assa# /m ;5ml 50.8 50.9 54.1 53.5 53.4 53.8 53.4 1.49 1.9% 1.50
April 2007

-ombined values :ean S"7 6S7 -. 95% 53.5 1.49 1.9% 1.40

53.6 53.0 53.4 53.9 54.3 54.0 53.8 1.44 1.9% 1.46

Slide 25 of 43

4pecificity "selectivity$
4peci;icity is the a'ility to assess une$ui"ocally the analyte in the presence o; components, =hich may 'e expecte! to 'e present. )ypically these might inclu!e impurities, !egra!ants an! excipients. excipients An example o; speci;icity criterion ;or an assay metho! is that the analyte pea9 =ill ha"e 'aseline chromatographic resolution o; at least ?.D minutes ;rom all other sample components 4ta'ility in!icating analytical metho!s shoul! al=ays 'e speci;ic.
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April 2007

!dentification 3 a special case

Diethylene glycol 0D*,1 in pae!iatric !osage ;orms has 'een implicate! as the causati"e agent in numerous !eaths since @FGH. )he "ictims =ere mainly chil!ren. #llustrati"e analytical issues o; in"estigation
% #& i!entity test =as a'le to !etect D*, at a'out ?D L=>= % )esting o; D*, in ,lycerol 0an! in Propylene ,lycol1 =as recommen!e! =ith a .OD 0sensiti"ity1 o; (.) D.@ L. 2or !etecting D*, at lo= le"els, ,+ seeme! pre;era'le. % )he assay =as the most rele"ant test 0accurate =ithin O D.?L1

#llustrati"e regulatory issues

% .egislation % , P

4peci;icity is an essential 'ut not su;;icient characteristic o; i!enti;ication

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Slide 27 of 43

April 2007

4pecificity "hypothetical figures and data$

3'0& c(#omato4#ams o+ 5a6 A'I #e+e#ence stan)a#)7 5!6 8'' an) 5c6 *"ace!o

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April 2007

4P0#! !#!T< 3 degradants

St#ess Initia" Aci) 'e#o:i)e A"" ot(e#s 9 5%6 A 100.0 99.3 99.8 100.0 ' #ity an4"e 0.040 0.105 0.725 0.045 0.120 1.040 0.060 0.110 0.690 %A ' #ity t(#es(o") 0.280 0.380 1.630 0.280 0.410 1.610 0.270 0.360 1.250 %A

T(e#e ;e#e no *ea2s in t(e *"ace!o c(#omato4#am at t(e #etention times o+ ne,i#a*ine 5%67 * met(y"*a#a!en 5M'6 an) *#o*y"*a#a!en 5''6 *ea2s S m o+ %7 M' an) '' *ea2 a#eas. T(e t(#ee in4#e)ients can !e assesse) in t(e *#esence o+ 5non< = e:*ecte)6 )e4#a)ants. T(e *ea2s a#e (omo4eneo s an) * #e. T(e met(o) is se"ecti,e7 s*eci+ic *an) sta!i"ity<in)icatin4
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Slide 29 of 43

April 2007

;OD> ;O' and 4/2

.imit o; 7uantitation 0.O71 .imit o; Detection 0.OD1 4ignal to (oise &atio 04(&1

*ea( = L<,

*ea( A L<7
(aselin e
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noise

Slide 30 of 43

April 2007

;OD and ;O' "hypothetical figures$

Injection 1 2 3 4 5 6 Mean STD RSD 6&onc. 5>4?m" 6&onc. 5%;?;
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Im* #ity 1 0.D 4176 3608 4196 4303 3932 5238 4242 548 12.9% 0.086 0.017 0.= 7235 8099 7950 8166 7847 8415 7952 402 5.1% 0.171 0.033

Im* #ity 2 0.D 3497 4258 3275 3464 4008 4702 3867 551 14.3% 0.107 0.019 0.= 7892 7791 8292 8050 8368 8284 8113 238 2.9% 0.214 0.039

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April 2007

;OD and ;O'

)he limit o; !etection 0.OD1 is !e;ine! as the lo=est concentration o; an analyte in a sample that can 'e !etecte!, not $uanti;ie!. #t is expresse! as a concentration at a speci;ie! signal : noise ratio 04(&1, usually 'et=een G an! ? : @. #n this stu!y, the .OD =as !etermine! to 'e D.DKE Pg>ml 0#mpurity @1 =ith a signal : noise ratio o; G.E : @ )he limit o; $uantitation 0.O71 is !e;ine! as the lo=est concentration o; an analyte in a sample that can 'e !etermine! =ith accepta'le precision an! accuracy un!er the state! operational con!itions o; the metho!. )he #+H has recommen!e! a signal : noise ratio 04(&1 o; @D:@. )he .O7 =as D.@H@ Pg>ml 0#mpurity @1 =ith a signal:noise ratio o; @@.G. )he &4D ;or six injections o; the .O7 solution =as M?L.
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Slide 32 of 43

April 2007

;inearity
Meas #e) mean Rea" mean

'#ecision

.inearity expresses !i;;erences in precision at !i;;erent points o; a gi"en range. A)he linearity o; an analytical proce!ure is its a'ility 0=ithin a gi"en range1 to o'tain test results, =hich are !irectly proportional to the concentration 0amount1 o; analyte in the sample.C

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April 2007

;inearity and range

5111 4111 Assay mean 4111 3111 0111 1 1 31 41 61 91 011 031 041 &oncent#ation7 %
Acceptance criterion( correlation coefficient should not be less than ?*,,,?
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# ! 46.034> ' 8.3994 6 ! 1.9999

3

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April 2007

;inearity and range

+oncentration range @.D%@.G mg>ml 0@D%@GDL o; the theoretical concentration in the test preparation, n5G1 &egression e$uation =as ;oun! 'y plotting the means o; pea9 area 0y1 against the analyte concentration 0x1 expresse! in L: y 5 GE.@?Bx < H.?FKB 0&? 5 D.FFFK1. )he regression coe;;icient !emonstrates an excellent relationship 'et=een pea9 area an! concentration o; analyte. )he analyte response is linear across @D<@GDL o; the target ne"irapine concentration.

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April 2007

2ange "minimum re%uirements$

Assay o; an AP# or a 2PP: O ?DL o; the test concentration. +ontent uni;ormity: O GDL o; the test concentration 0unless a =i!er
more appropriate range, 'ase! on the nature o; the !osage ;orm 0e.g., metere! !ose inhalers1, is justi;ie!1.

Dissolution testing: O ?D L o"er the speci;ie! range. #mpurity: ;rom the reporting le"el o; an impurity to @?DL o; the speci;ication. 03nusually potent or toxic impurities, .OD an! .O7 shoul! 'e
commensurate =ith #+H re$uirement.1

#; assay an! purity are per;orme! together as one test an! only a @DDL stan!ar! is use!, linearity shoul! co"er the range ;rom the reporting le"el o; the impurities to @?DL o; the assay speci;ication

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April 2007

4tability of analytical solution

4ta'ility 0o; the analytical solution1 expresses variation o; the measure! mean as a ;unction o; time.

Sta!i"ity

:easured means

N@ Q 2irst measurements N?, NG, NB, Qn 4eries o; measurements o; the same sample wit$in a relativel# s$ort period of time.

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April 2007

4tability of test analytical solution

Im* #ity<1 Time in (o #s 0 1 2 3 4 5 10 15 20 25
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A#ea 72079 71574 71740 71960 72352 71573 72322 72310 72312 72670
April 2007

Di++e#ence 0.7% 0.5% 0.2% 0.4%< 0.7% 0.3%<

An analytical solution prepare! ;rom (e"irapine JDmg>Jml Oral 4uspension =as spi9e! =ith #mpurity<@ at speci;ication le"el an! store! in a cappe! "olumetric ;las9 on a la'oratory 'ench at uncontrolle! room temperature un!er normal lighting con!itions ;or ?J hours.

+onclusion: the sta'ility o; the 0.3%< analytical solution o; #mpurity<@ is 0.3%< not a source o; "ariation.
0.8%<

Slide 38 of 43

4ensitivity and robustness

Slide 39 of 43

April 2007

2obustness
t6 :et$od parameter S"* )low @avelen t$ ?ariation of mobile p$ase -olumn temperature pB
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?ariation ' 01%' 01% 5nm' 5nmA 3%' 3%A 5o-' 5o-A 1.4' 1.4A

.mpurit# 0 1.94 1.94 1.94 1.93 1.94 1.91 1.94 1.93 1.94 1.94 1.94

.mpurit# 3 0.91 0.90 0.93 0.90 0.90 0.99 0.86 0.91 0.90 0.90 0.91

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April 2007

Methods for cleaning validation

etho! ;or assay an! relate! su'stances use! in sta'ility stu!ies o; AP# an! 2PP
% 4peci;icity 0in samples ta9en ;rom a cleaning assessment1 % .inearity o; response 0;rom JDL o; the cleaning limit to @Dx this concentrationR &? S D.FFDDR 1 % Precision
Repeatability (RSD 5%) , intermediate precision [ruggedness (USP) , and reproducibility

% .imits o; !etection an! $uantitation % Accuracy or reco"ery ;rom rinsate 0S KDL1, s=a's 0S FDL1, an! process sur;ace 0S HDL1 % &ange 0lo=est le"el is at least ?x higher than .O71
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April 2007

Main Points Again

Analytical proce!ures play a critical role in pharmaceutical e$ui"alence an! ris9 assessment > management:
% esta'lishment o; pro!uct<speci;ic acceptance criteria, an! % sta'ility o; AP#s an! 2PPs.

6ali!ation shoul! !emonstrate that the analytical proce!ure is suita'le ;or its intente! purpose. HP.+ systems an! metho! "ali!ation !eser"es special attention !uring the assessment o; !ossiers ;or pre$uali;ication.
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April 2007

T@A/A <O5

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April 2007