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Birgit Schmauser | April 20081 |
Pharmaceutical Development with Focus
on Paediatric formulations
Pharmaceutical Development with Focus
on Paediatric formulations
WHO/FIP Training Workshop
Hyatt Regency Hotel
Sahar Airport Road
Andheri East, Mumbai, India
28 April 2008 2 May 2008
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Birgit Schmauser | April 20082 |
Analytical Method DevelopmentAnalytical Method Development
Presented by:Presented by:
Birgit Schmauser, PhDBirgit Schmauser, PhD
Federal Institute for DrugsFederal Institute for Drugs
and Medical Devices (BfArM)and Medical Devices (BfArM)
[email protected]@bfarm.de
http://www.travelphoto.net/photos/pictures/indien/in10.jpg8/2/2019 Analy Meth Dev
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Birgit Schmauser | April 20083 |
Analytical Method DevelopmentAnalytical Method Development
In this presentation:In this presentation:
Standards in developing analytical methods forStandards in developing analytical methods for
Originator and multisource generic FPPsOriginator and multisource generic FPPs
SpecificationsSpecifications
StabilityStability
Parallel development of analytical methods forParallel development of analytical methods for
cleaning validationcleaning validation
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Birgit Schmauser | April 20084 |
Analytical Method DevelopmentAnalytical Method Development
OriginatorOriginator,, First-timeFirst-time GenericGeneric and Multisource Genericand Multisource GenericOriginatorOriginator First-timeFirst-time GenericGeneric Multisource GenericMultisource Generic
API qualityAPI qualitystandardsstandards
OriginatorsOriginatorsspecificationsspecifications
Information fromInformation fromregulatory agenciesregulatory agencies(publicly available) &(publicly available) &
literature dataliterature data
PharmacopoeiasPharmacopoeias
FPP qualityFPP qualitystandardsstandards
OriginatorsOriginatorsspecificationsspecifications
Information fromInformation fromregulatory agenciesregulatory agencies(publicly available) &(publicly available) &literature dataliterature data
PharmacopoeiasPharmacopoeias
AnalyticalAnalytical
methodsmethods
EstablishEstablish identity,identity,
potency, purity of APIpotency, purity of APIand FPP byand FPP byin-house methodsin-house methods
DeriveDerive identity, potency,identity, potency,
purity of API and FPP bypurity of API and FPP byin house methodsin house methods
VerifyVerify identity, potency,identity, potency,
purity of API and FPP bypurity of API and FPP bypharmacopoeial methodspharmacopoeial methodsand in-house methodsand in-house methods
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Birgit Schmauser | April 20085 |
Analytical Method DevelopmentAnalytical Method Development
HPLC-method to assay potency and purity risk assessmentHPLC-method to assay potency and purity risk assessmentOriginatorOriginator First-timeFirst-time GenericGeneric Multisource GenericMultisource Generic
SelectivelySelectively screenscreen/detect any/detect anyimpurity or degradantimpurity or degradantEstablishEstablish potencypotency
IdentifyIdentify impurities/degradantsimpurities/degradants
CharacteriseCharacterise allallimpurities/degradantsimp
urities/degradants CalculateCalculateResponse factorsResponse factors(qualification by clinical use)(qualification by clinical use)
DeriveDerive impurities/degradants fromimpurities/degradants fromOriginatorOriginatorCharacterize in-houseCharacterize in-houseimpurities/degradantsimpurities/degradantsCalculate response factorsCalculate response factors
Verify impurities fromVerify impurities fromPharmacopoeiaPharmacopoeiaCharacterise in-houseCharacterise in-houseimpurities/degradantsimpurities/degradants(Response factors)(Response factors)
EstablishEstablish reference materialsreference materials ExtractExtract (&(& reproducereproduce) reference) referencematerialsmaterials
Use pharmacopoeial referenceUse pharmacopoeial referencematerialsmaterials
AdaptAdapt to routine useto routine use AdaptAdapt/modify to/for routine use/modify to/for routine use Implement for routine useImplement for routine use
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Birgit Schmauser | April 20086 |
Analytical Method DevelopmentAnalytical Method Development
Interchangeability (IC) ofInterchangeability (IC) ofmultisource generic FPPsmultisource generic FPPs((Essential similarityEssential similarity with Innovator FPP)with Innovator FPP)
PharmaceuticalPharmaceutical ++ BioequivalenceBioequivalenceEquivalenceEquivalence
IC =IC = PEPE ++ BEBE
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Birgit Schmauser | April 20087 |
Analytical Method DevelopmentAnalytical Method Development
Pharmaceutical equivalencePharmaceutical equivalence FPPs meet theFPPs meet the samesame ororcomparablecomparable standardsstandards byby
use of equivalent analytical methodsuse of eq
uivalent analytical methods
SameSame APIAPI ((chemicalchemical andand physicalphysical
equivalence)equivalence) SameSame dosage formdosage form andand route ofroute of
administrationadministration
SameSame strengthstrength
ComparableComparable labelinglabeling Equivalence inEquivalence in pharmaceutical developmentpharmaceutical development
Equivalence inEquivalence in stabilitystability
Equivalence inEquivalence in manufacture (WHO-GMP)manufacture (WHO-GMP)
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Analytical Method DevelopmentAnalytical Method Development
Prequalification requirementsPrequalification requirements Validation of analytical methods is aValidation of analytical methods is a prerequisiteprerequisite forfor
prequalification of product dossiersprequalification of product dossiers
Non-compendial APIs and FPPsNon-compendial APIs and FPPs are tested withare tested with methodsmethodsdeveloped by the manufacturerdeveloped by the manufacturer
ForFor compendial APIs and FPPs the applicability ofcompendial APIs and FPPs the applicability ofpharmacopoeial methodspharmacopoeial methods to particular productsto particular products mustmust
be demonstrated (verification)be demonstrated (verification)
Analytical methods must be developed and validatedAnalytical methods must be developed and validated
according to TRS 823,according to TRS 823, Annex 5,Annex 5, Validation of analyticalValidation of analytical
procedures used in the examination of pharmaceuticalprocedures used in the examination of pharmaceuticalmaterialsmaterials; ICH Q2 (R1); ICH Q2 (R1)
To be used within GLP and GMP environmentsTo be used within GLP and GMP environments
8/2/2019 Analy Meth Dev
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Analytical Method DevelopmentAnalytical Method Development
CLINICALCLINICAL PHARMACEUTICALPHARMACEUTICAL METHODSMETHODS
AtAt initialinitial phase of pharmaceutical developmentphase of pharmaceutical development
ToTo determinedeterminebioavailabilitybioavailability in healthyin healthyvolunteersvolunteers
To develop a stable andTo develop a stable andreproducible formulation for thereproducible formulation for themanufacture of bioequivalence,manufacture of bioequivalence,
dissolution, stability and pilot-dissolution, stability and pilot-scale validation batchesscale validation batches
To understand theTo understand the profileprofile of relatedof relatedsubstances and to studysubstances and to study stabilitystability
To startTo start measuringmeasuring the impact ofthe impact ofkeykey
productproduct andand manufacturing processmanufacturing processparametersparameters onon consistentconsistent FPP qualityFPP quality
AtAt advancedadvanced phase of pharmaceutical developmentphase of pharmaceutical development
To proveTo prove bioequivalencebioequivalenceafteraftercritical variationscritical variations totothe prequalified dossierthe prequalified dossier
ToTo optimiseoptimise,, scale-upscale-up andand transfertransferaa stablestable andand controlledcontrolledmanufacturing process for themanufacturing process for theprequalification productprequalification product
To beTo be robust, transferable, accuraterobust, transferable, accurateandand preciseprecise for specification setting,for specification setting,stability assessment and QC release ofstability assessment and QC release ofprequalified product batchesprequalified product batches
Use of analytical methods -Use of analytical methods - genericsgenerics
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Birgit Schmauser | April 200810 |
Analytical Method DevelopmentAnalytical Method Development
PrerequisitesPrerequisites for analytical method validationfor analytical method validation Six Ms
Quality of theQuality of the
analytical methodanalytical method
MManan MMachineachine
qualifiedqualified
calibratedcalibrated
robustrobust
qualifiedqualified
MMethodsethods
suitablesuitable
characterisedcharacterised
documenteddocumented
MMilieuilieuMMaterialaterial MManagementanagement
QualityQuality
ReferenceReference
standardsstandards
Tempe-Tempe-raturerature
AnalystsAnalysts
supportsupport
skilledskilled
HumidityHumidity
VibrationsVibrations TimeTime
SuppliesSupplies
Irradi-Irradi-
ationsations
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Birgit Schmauser | April 200811 |
Analytical Method DevelopmentAnalytical Method Development
Method development life cycleMethod development life cycle
Planning
Development and Validation Policy
Objectives/Requirements of Method
Information GatheringResource Gathering
Method developmentInitital Method Development
Pre-Validation Evaluation
Method Optimization
Robustness
System Suitability
Development
Plan
Project
Customer Evaluation
TestingValidation Experiments
Method Transfer
ExperimentsFiled Method in Use
Periodically
Monitoring/Reviewof Methods
in Control LabsFrom: Analytical Chemistry in a GMP Environment. Edited by J.M. Miller and J.B. Crowther, ISBN 0-471-31431-5, Wiley & Sons IncFrom: Analytical Chemistry in a GMP Environment. Edited by J.M. Miller and J.B. Crowther, ISBN 0-471-31431-5, Wiley & Sons Inc..
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Birgit Schmauser | April 200812 |
Analytical Method DevelopmentAnalytical Method Development
Validation should verify the suitability of anValidation should verify the suitability of ananalytical methodanalytical method for its intended purposefor its intended purpose
Validation should be founded onValidation should be founded on methodmethod
development performed beforehanddevelopment performed beforehand thatthatsuggest thesuggest the suitability and robustnesssuitability and robustness of theof themethodmethod
Validation may be performed in different waysValidation may be performed in different ways(individual purpose) according to(individual purpose) according to commoncommonstandardsstandards
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Birgit Schmauser | April 200813 |
Validation protocolValidation protocol Method principle / objectiveMethod principle / objective
Listing of responsibilitiesListing of responsibilities
Laboratories involved and their role in the validationLaboratories involved and their role in the validation
Method categorizationMethod categorization
List of reagents (including test lots) and standardsList of reagents (including test lots) and standards
Test procedures to evaluate each validation parameter and proposedTest procedures to evaluate each validation parameter and proposedacceptance criteriaacceptance criteria
Plan or procedure when acceptance criteria are not metPlan or procedure when acceptance criteria are not met
Requirements for the final reportRequirements for the final report
The validation process cannot proceed until the protocol and allThe validation process cannot proceed until the protocol and allparties involved approve the acceptance criteriaparties involved approve the acceptance criteria
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Birgit Schmauser | April 200814 |
Analytical Method DevelopmentAnalytical Method Development
Innovator versus GenericsInnovator versus Generics
InnovatorInnovator GenericsGenerics
R & D on APIR & D on API ++ --
Preclinical trialsPreclinical trials ++ --
Clinical trials phase I and IIClinical trials phase I and II Method validationMethod validation
summarysummary--
Clinical trials phase IIIClinical trials phase III Method validationMethod validationcompletedcompleted
--
Post marketing phase IVPost marketing phase IV Validated methodsValidated methods --
Entering of Generics; PharmaceuticalEntering of Generics; Pharmaceutical
development, Comparability withdevelopment, Comparability withInnovatorInnovator
Validated methodsValidated methods Validated methods:Validated methods:
GMP and GLPGMP and GLP
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Birgit Schmauser | April 200815 |
Analytical Method DevelopmentAnalytical Method Development
Validation CharacteristicsValidation CharacteristicsIdentificationIdentification ImpuritiesImpurities AssayAssay
quantitativequantitative limitlimit
AccuracyAccuracy -- ++ -- ++
PrecisionPrecision -- ++ -- ++SpecificitySpecificity ++ ++ ++ ++
Detection LimitDetection Limit -- -- ++ --
Quantitation LimitQuantitation Limit -- ++ -- --
LinearityLinearity -- ++ -- ++
RangeRange -- ++ -- ++
RobustnessRobustness ++ ++ ++ ++
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Birgit Schmauser | April 200816 |
Analytical Method DevelopmentAnalytical Method Development
Accuracy and precisionAccuracy and precision
Accurate &Accurate &
preciseprecise Accurate &Accurate &impreciseimprecise
InaccurateInaccurate&&
preciseprecise Inaccurate & impreciseInaccurate & imprecise
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Birgit Schmauser | April 200817 |
Analytical Method DevelopmentAnalytical Method Development
PrecisionPrecision Expresses theExpresses the closeness of agreementcloseness of agreement between a series ofbetween a series of
measurements obtained from multiple sampling of the samemeasurements obtained from multiple sampling of the same
homogenous samplehomogenous sample
Is usually expressed as theIs usually expressed as the standard deviation (S),standard deviation (S), variancevariance (S(S22))
ororcoefficient of variationcoefficient of variation (RSD)(RSD) of a series of measurementsof a series of measurements
Precision may be considered at three levelsPrecision may be considered at three levels
RepeatabilityRepeatability(intra-assay precision)(intra-assay precision)
Intermediate PrecisionIntermediate Precision(variability within a(variability within a
laboratory)laboratory) ReproducibilityReproducibility(precision between laboratories)(precision between laboratories)
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Birgit Schmauser | April 200818 |
Analytical Method DevelopmentAnalytical Method Development
Normal distribution,Normal distribution, probability function [P(x)]probability function [P(x)]
and confidence interval [CI]and confidence interval [CI] Probability (P)Probability (P), that measurements from a normal distribution fall within [-x, that measurements from a normal distribution fall within [-xnn, +x, +xnn]]
for xfor xnn = n= n is described by theis described by the erferf-function-function ((= mean)mean):
An interval of 3An interval of 3 coverscovers 99.73% of values99.73% of values
Numberoftimeseachvalue
occursxxnn PP
0.68268950.6826895
22 0.95449970.9544997
33 0.99730020.9973002
44 0.99993660.9999366
55 0.99999940.9999994
Values23 2 3
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Birgit Schmauser | April 200819 |
Analytical Method DevelopmentAnalytical Method Development
Normal distribution, probability function [P(x)] andNormal distribution, probability function [P(x)] and
confidenceconfidence
interval [CI]interval [CI]
Probability-PProbability-P Confidence interval [CI]Confidence interval [CI]
centered around the mean []centered around the mean []
in units of sigma [in units of sigma [] described by] described byinverseinverse erferf-function:-function:
A CI of 95% includes valuesA CI of 95% includes values
1.95 1.95 around the meanaround the mean
PP xxpp
0.8000.800 1.281551.28155
0.9000.900 1.644851.64485
0.9500.950 1.959961.95996
0.9950.995 2.575832.57583
0.9990.999 3.290533.29053
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Birgit Schmauser | April 200820 |
Analytical Method DevelopmentAnalytical Method Development
Relationship of variability, probability and reliability of dataRelationship of variability, probability and reliability of data High variability of data (largeHigh variability of data (large ) generate large confidence intervals and) generate large confidence intervals and
thus lower the reliability of the meanthus lower the reliability of the mean
Low variability of data (smallLow variability of data (small )) generate small confidence intervals andgenerate small confidence intervals andthus increase the reliability of the meanthus increase the reliability of the mean
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Birgit Schmauser | April 200821 |
Analytical Method DevelopmentAnalytical Method Development
RepeatabilityRepeatability Six replicate sample preparation steps from a homogenously prepared tabletSix replicate sample preparation steps from a homogenously prepared tablet
mixture (nominal value of API 150 mg)mixture (nominal value of API 150 mg)
InjectionInjection Peak areaPeak area AssayAssay
11 173865173865 147.10 mg/98.06%147.10 mg/98.06%
22 174926174926 148.00 mg/98.66%148.00 mg/98.66%
33 172933172933 146.32 mg/97.54%146.32 mg/97.54%
44 175011175011 148.08 mg/98.72%148.08 mg/98.72%
55 179557179557 151.95 mg/101.30%151.95 mg/101.30%
66 176425176425 149.28 mg/99.52%149.28 mg/99.52%
MeanMean 175453175453 148.45 mg/98.96%148.45 mg/98.96%
SD (SD ()) 23292329 1.98 mg/1.32%1.98 mg/1.32%
RSDRSD 1.32%1.32% 1.32%1.32%
MeanMean 3 SD = 3 SD =Confidence interval of 99.73%Confidence interval of 99.73%
98.9698.963x1.32% =3x1.32% = 95%95% - 102.92%- 102.92%
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Birgit Schmauser | April 200822 |
Analytical Method DevelopmentAnalytical Method Development
Intermediate precisionIntermediate precision Expresses within-laboratories variations (different days, different analysts,Expresses within-laboratories variations (different days, different analysts,
different equipment etc.)different equipment etc.)
InjectionInjection Peak areaPeak areaanalyst 1analyst 1
Peak areaPeak areaanalyst 2analyst 2
Peak areaPeak areaanalyst 3analyst 3
11 173865173865 175656175656 17796517796522 174926174926 175878175878 178556178556
33 172933172933 176004176004 177342177342
44 175011175011 176344176344 178011178011
55 179557179557 175332175332 179466179466
66 176425176425 174959174959 179688179688
MeanMean 175453175453 175695175695 178504178504SD (SD ()) 23292329 495495 918918
RSDRSD 1.32%1.32% 0.28%0.28% 0.51%0.51%
Analyst 1: 98.96%Analyst 1: 98.96% 3 x 1.32% 3 x 1.32%
Analyst 2: 99.12%Analyst 2: 99.12% 3 x 0.28 3 x 0.28
Analyst 3: 100.70%Analyst 3: 100.70% 3 x 0.51 3 x 0.51
Average of 3 analystsAverage of 3 analysts 3SD 3SD::
95%95% - 102.23%- 102.23%
MeanMean 3 SD: (177252 3 SD: (177252
100%100%))
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Birgit Schmauser | April 200823 |
Analytical Method DevelopmentAnalytical Method Development
ReproducibilityReproducibility
Expresses the precision between laboratoriesExpresses the precision between laboratories
Collaborative studies, usually applied toCollaborative studies, usually applied to
standardisation of methodologystandardisation of methodology
Transfer of technologyTransfer of technology
Compendial methodsCompendial methods
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Birgit Schmauser | April 200824 |
Analytical Method DevelopmentAnalytical Method Development
AccuracyAccuracy Expresses theExpresses the closeness of agreementcloseness of agreement betweenbetween the value which isthe value which is
accepted either as a conventionalaccepted either as a conventional true valuetrue value ororan acceptedan accepted
reference valuereference value and theand the value foundvalue found
Sometimes referred to as Sometimes referred to as TRUENESSTRUENESS
truetruemeanmean
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Birgit Schmauser | April 200825 |
Analytical Method DevelopmentAnalytical Method Development
To find out whether a method is accurate:To find out whether a method is accurate: Drug substance (assay)Drug substance (assay)
Application of the method to an analyte of known purity (e.g. referenceApplication of the method to an analyte of known purity (e.g. referencesubstance)substance)
Comparison of the results of one method with those of a second well-Comparison of the results of one method with those of a second well-
characterised method (accuracy known)characterised method (accuracy known)
Drug product (assay)Drug product (assay) Application of the method to synthetic mixtures of the drug product componentApplication of the method to synthetic mixtures of the drug product component
to whichto which known quantitiesknown quantities of the analyte have been addedof the analyte have been added
Drug product may exceptionally be used as matrixDrug product may exceptionally be used as matrix
Drug substance/Drug product (Impurities)Drug substance/Drug product (Impurities) Application of the method to samples spiked withApplication of the method to samples spiked with known amountsknown amounts of impuritiesof impurities
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Birgit Schmauser | April 200826 |
Analytical Method DevelopmentAnalytical Method Development
AccuracyAccuracy:: Application of the method to synthetic mixtures of theApplication of the method to synthetic mixtures of the
drug product componentsdrug product components
to which known quantitiesto which known quantities
of the analyteof the analyte
have been addedhave been added
RecoveryRecovery reducedreduced
byby ~10 15%~10 15%
From: Analytical Method Validation and Instrument Performance Verification, Edited byFrom: Analytical Method Validation and Instrument Performance Verification, Edited byChung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &Chung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &SonsSons
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Birgit Schmauser | April 200828 |
Analytical Method DevelopmentAnalytical Method Development
SpecificitySpecificity Is the ability to assess unequivocally the analyte in the presence of componentsIs the ability to assess unequivocally the analyte in the presence of components
which may be expected to be present (impurities, degradants, matrix)which may be expected to be present (impurities, degradants, matrix)
Identity testingIdentity testing
To ensure the identity of an analyteTo ensure the identity of an analyte
Purity testingPurity testing
To ensure accurate statement on the content of impurities of an analyteTo ensure accurate statement on the content of impurities of an analyte
AssayAssay
To allow an accurate statement on the content of an analyte in a sampleTo allow an accurate statement on the content of an analyte in a sample
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Birgit Schmauser | April 200829 |
Analytical Method DevelopmentAnalytical Method Development
Specificity:Specificity:Overlay chromatogram of an impurity solution with aOverlay chromatogram of an impurity solution with asample solutionsample solution
From: Analytical Method Validation and Instrument Performance Verification, Edited byFrom: Analytical Method Validation and Instrument Performance Verification, Edited byChung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &Chung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &SonsSons
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Birgit Schmauser | April 200830 |
Analytical Method DevelopmentAnalytical Method Development
Specificity andSpecificity and stabilitystability Stress stability testing to ensure theStress stability testing to ensure the stability indicating potentialstability indicating potential of anof an
analytical methodanalytical method Apply diverse stress factors to the APIApply diverse stress factors to the API
Apply diverse stress factors to the FPPApply diverse stress factors to the FPP
Stress conditions: e.g. Supplement 2 of Generic Guideline;Stress conditions: e.g. Supplement 2 of Generic Guideline;TRS 929, Annex 5TRS 929, Annex 5
Assure that the API can be assessed specifically in the presence ofAssure that the API can be assessed specifically in the presence ofknown and unknown (generated by stress) impuritiesknown and unknown (generated by stress) impurities
Assure that known impurities/degradants can be specifically assessedAssure that known impurities/degradants can be specifically assessedin the presence of further degradantsin the presence of further degradants
ByBy peak purity assessmentpeak purity assessment and (overlay of)and (overlay of) chromatogramschromatograms
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Birgit Schmauser | April 200831 |
Analytical Method DevelopmentAnalytical Method Development
StressStress stability studies versusstability studies versus forced degradationforced degradation studiesstudiesStressStress
parameterparameterForced degradationForced degradation Stress stabilityStress stability
(5 15% decomposition)(5 15% decomposition)
AcidAcid 0.2 ml0.2 ml 1N HCl1N HCl / 5 ml API-solution / 3h,/ 5 ml API-solution / 3h,6h, 12h, 24h7d (RT & 60C)6h, 12h, 24h7d (RT & 60C)
pHpH 2 (2 weeks) 2 (2 weeks)
BaseBase 0.2 ml0.2 ml 1N NaOH1N NaOH / 5 ml API-solution // 5 ml API-solution /3h, 6h, 12h, 24h7d (RT & 60C)3h, 6h, 12h, 24h7d (RT & 60C)
pHpH 10 (2 weeks) 10 (2 weeks)
HH22OO22 / Oxygen/ Oxygen 0.2 ml 5% or0.2 ml 5% or35%35% HH22OO22/ 5 ml API-/ 5 ml API-
solution (RT, to 7d & 60C, 3h)solution (RT, to 7d & 60C, 3h)
1 g/ml oxygen bubbled through (8 hours)1 g/ml oxygen bubbled through (8 hours)0.1 2% H0.1 2% H22OO22 (24 hours)(24 hours)
HeatHeat 60C60C / 5 ml solution (3h, 6h7d)/ 5 ml solution (3h, 6h7d) --
HeatHeat 105 C105 C / solid API (1d and 7d)/ solid API (1d and 7d) 60C (4 weeks)60C (4 weeks)
UV or LightUV or Light 365 nm or white fluorescent light / solid365 nm or white fluorescent light / solidAPI (1d and 7d)API (1d and 7d)
HumidityHumidity -- 50C / 80% RH (4 weeks)50C / 80% RH (4 weeks)
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Birgit Schmauser | April 200832 |
Analytical Method DevelopmentAnalytical Method Development
Limit of Detection (LOD, DL)Limit of Detection (LOD, DL) The LOD of an analytical procedure is the lowest amount of analyte in sampleThe LOD of an analytical procedure is the lowest amount of analyte in sample
which can bewhich can be detected but not necessarily quantitated as an exact valuedetected but not necessarily quantitated as an exact value
Determination is usually based onDetermination is usually based on
Signal to noise ratio (~3:1) (Signal to noise ratio (~3:1) (baseline noisebaseline noise))oror
Standard deviation of response (Standard deviation of response () and Slope (S)) and Slope (S)
3.33.3 /S/S
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Birgit Schmauser | April 200833 |
Analytical Method DevelopmentAnalytical Method Development
Limit of Quantitation (LOQ, QL)Limit of Quantitation (LOQ, QL) The LOQ is the lowest amount of analyte in a sample which can beThe LOQ is the lowest amount of analyte in a sample which can be
quantitativelyquantitatively determined with suitable precision and accuracydetermined with suitable precision and accuracy
The quantitation limit is used particularly for theThe quantitation limit is used particularly for thedetermination of impurities and/or degradationdetermination of impurities and/or degradation
productsproducts
Determination is usually based onDetermination is usually based on
Signal to noise ratio (~10:1) (Signal to noise ratio (~10:1) (baseline noisebaseline noise))
oror Standard deviation of response (Standard deviation of response () and Slope (S)) and Slope (S)
1010 //SS
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Birgit Schmauser | April 200834 |
Analytical Method DevelopmentAnalytical Method Development
NoiseNoise
LODLODSignal toSignal to NoiseNoise = 3:= 3:11
LOQLOQ
Signal toSignal to NoiseNoise = 10:= 10:11
LOD, LOQ and Signal to Noise Ratio (SNRLOD, LOQ and Signal to Noise Ratio (SNR))
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Birgit Schmauser | April 200835 |
Analytical Method DevelopmentAnalytical Method Development
LOQLOQ Quantitation by SNR is acceptedQuantitation by SNR is accepted
Quantitation by Standard deviation of response (Quantitation by Standard deviation of response () and Slope (S)) and Slope (S)
(10(10 //S) is more adequate as it involves theS) is more adequate as it involves the responseresponse of the actualof the actual
analyteanalyte
Best to calculate in the region close to y-interceptBest to calculate in the region close to y-intercept
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Birgit Schmauser | April 200836 |
Analytical Method DevelopmentAnalytical Method Development
LOQ and impuritiesLOQ and impurities
In determination of impurities in APIs and FPPs the LOQ should beIn determination of impurities in APIs and FPPs the LOQ should be
determineddetermined in the presence of APIin the presence of API
LOQ should beLOQ should be NMT reporting levelNMT reporting level
LOQ should be givenLOQ should be given relativerelative to the test concentrationto the test concentrationof APIof API
Specificity of impurity determination should always be demonstratedSpecificity of impurity determination should always be demonstrated
in the presence of APIin the presence of API at API specification levelsat API specification levels
Spiking of test concentration (API/FPP) with impurities atSpiking of test concentration (API/FPP) with impurities atlevels of their specification rangelevels of their specification range
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Birgit Schmauser | April 200837 |
Analytical Method DevelopmentAnalytical Method Development
SpikingSpiking API test concentration (normalised)API test concentration (normalised)
0.1 mg/ml (100%)0.1 mg/ml (100%)
Impurity spiking concentrationsImpurity spiking concentrations
0.001 mg/ml (1%) specification limit0.001 mg/ml (1%) specification limit 0.0001 mg/ml (0.1%)0.0001 mg/ml (0.1%) limit of quantitation limit of quantitation(minimum requirement)(minimum requirement)
API at test concentrationsAPI at test concentrations
API below test concentrationsAPI below test concentrations
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Birgit Schmauser | April 200838 |
Analytical Method DevelopmentAnalytical Method Development
LinearityLinearityof an analytical procedure is its ability (of an analytical procedure is its ability (within a givenwithin a given rangerange))to obtain testto obtain test resultsresultswhich are directlywhich are directly proportional to theproportional to the
concentrationconcentration (amount) of analyte in the sample(amount) of analyte in the sample
If there is aIf there is a linearlinearrelationshiprelationship test results should be evaluatedtest results should be evaluatedby appropriateby appropriate statistical methodsstatistical methods
Correlation coefficient (r)Correlation coefficient (r)
Y-interceptY-intercept
Slope of regression lineSlope of regression line
Residual sum of squaresResidual sum of squares
PLOT OF THE DATAPLOT OF THE DATA
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Birgit Schmauser | April 200839 |
Analytical Method DevelopmentAnalytical Method Development
Usual acceptance criteria for a linear calibrationUsual acceptance criteria for a linear calibrationcurvecurve r > 0.999r > 0.999;; y-intercept a < 0 to 5% of target concentrationy-intercept a < 0 to 5% of target concentration
RSD (wrt calibration curve) < 1.5-2%RSD (wrt calibration curve) < 1.5-2%
r > 0.997 r < 0.997
From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung ChowFrom: Analytical Method Validation and Instrument Performance Verification, Edited by Chung ChowChan,Herman Lam, Y.C. Lee andChan,Herman Lam, Y.C. Lee andXue-Ming Zhang, ISBN 0-471-25953-5, Wiley & SonsXue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons
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Birgit Schmauser | April 200840 |
Analytical Method DevelopmentAnalytical Method Development
RangeRange
TheThe rangerange of an analytical procedure is the intervalof an analytical procedure is the interval
between the upper and lower concentration (amounts) ofbetween the upper and lower concentration (amounts) of
analyte in the sample for which it has been demonstratedanalyte in the sample for which it has been demonstrated
that the analytical procedure has athat the analytical procedure has a suitable level ofsuitable level of
precision, accuracy and linearityprecision, accuracy and linearity
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Birgit Schmauser | April 200841 |
Analytical Method DevelopmentAnalytical Method Development
RangeRange AssayAssay 80 to 120%80 to 120% of test concentrationof test concentration
Content uniformityContent uniformity
70 to 130%70 to 130% of test concentrationof test concentration
DissolutionDissolution Q-20% to 120%Q-20% to 120%
ImpuritiesImpurities
Reporting level 120% of specification limit (with respect toReporting level 120% of specification limit (with respect totest concentration of API)test concentration of API)
Assay & ImpuritiesAssay & Impurities
Reporting level to 120% of assay specificationReporting level to 120% of assay specification
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Birgit Schmauser | April 200842 |
Analytical Method DevelopmentAnalytical Method Development
Linearity is limited to 150%of shelf life specification of impuritiesLinearity is limited to 150%of shelf life specification of impurities
Test concentration can beTest concentration can be
used to determine impuritiesused to determine impurities
To determine drug substanceTo determine drug substance(assay) the test concentration(assay) the test concentration
must be dilutedmust be diluted
The range is 0 ~ 150% ofThe range is 0 ~ 150% of
impurity specificationimpurity specification
From: Analytical Method Validation and Instrument Performance Verification, Edited byFrom: Analytical Method Validation and Instrument Performance Verification, Edited byChung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &Chung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley &SonsSons
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Birgit Schmauser | April 200843 |
Analytical Method DevelopmentAnalytical Method Development
RobustnessRobustness Robustness of an analytical procedure should showRobustness of an analytical procedure should show
thethe reliabilityreliability of an analysisof an analysis with respect towith respect to deliberatedeliberatevariations in method parametersvariations in method parameters
The evaluation of robustness should be consideredThe evaluation of robustness should be consideredduring theduring the development phasedevelopment phase
If measurements areIf measurements are susceptiblesusceptible to variations into variations inanalytical conditions theanalytical conditions the analytical conditions shouldanalytical conditions shouldbe suitably controlledbe suitably controlled or aor a precautionary statementprecautionary statementshould be included in the procedureshould be included in the procedure
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Birgit Schmauser | April 200844 |
Analytical Method DevelopmentAnalytical Method Development
Influence of buffer pH and buffer concentration in mobile phase onInfluence of buffer pH and buffer concentration in mobile phase onretention times of API and impuritiesretention times of API and impurities
Conclusion: The buffer composition should be maintained in a range ofConclusion: The buffer composition should be maintained in a range of
8585 0.5% 0.5% Missing:Missing: Acceptance criterion for maximal deviation of retention time should beAcceptance criterion for maximal deviation of retention time should be
defined unless justifieddefined unless justified
APIAPI Impurity AImpurity A Impurity BImpurity B Impurity CImpurity C
As isAs is 10.4610.46 3.863.86 7.437.43 8.268.26
buffer pH 5.9buffer pH 5.9 10.4510.45 3.943.94 7.517.51 8.388.38
buffer pH 6.9buffer pH 6.9 10.4610.46 3.943.94 7.497.49 8.348.34
Buffer conc. 83%Buffer conc. 83% 7.847.84 3.433.43 6.166.16 6.666.66
Buffer conc. 87%Buffer conc. 87% 15.2615.26 4.774.77 9.619.61 11.1811.18
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Birgit Schmauser | April 200845 |
Analytical Method DevelopmentAnalytical Method Development
System suitability testingSystem suitability testing
Based on the concept that equipment, electronics,Based on the concept that equipment, electronics,
analytical operations and samples to be analysedanalytical operations and samples to be analysed
constitute an integral systemconstitute an integral system that can be evaluated asthat can be evaluated as
suchsuch
Suitability parameters are established for each analyticalSuitability parameters are established for each analytical
procedureprocedureindividuallyindividually
Depend on theDepend on the type of analytical proceduretype of analytical procedure
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Birgit Schmauser | April 200846 |
Analytical Method DevelopmentAnalytical Method Development
MethodMethod stabilitystability System suitabilitySystem suitability over timeover time
Sample solution stabilitySample solution stability A solution of stavudine is stable for ~ 2 h, then it starts to degradeA solution of stavudine is stable for ~ 2 h, then it starts to degrade
to thymineto thymine
Impurity-spiked sample solution stabilityImpurity-spiked sample solution stabilityA solution containing stavudine spiked with its impurityA solution containing stavudine spiked with its impurity
thyminethyminedoes not allow to clearly distinguish betweendoes not allow to clearly distinguish betweendegradation and spikedegradation and spikeA solution containing stavudine of a FPP-stability sampleA solution containing stavudine of a FPP-stability sample
solution does not allow to clearly distinguish between FPP-solution does not allow to clearly distinguish between FPP-stability degradation and sample solution degradationstability degradation and sample solution degradation
Should be analysed immediatelyShould be analysed immediately
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Birgit Schmauser | April 200847 |
Analytical Method DevelopmentAnalytical Method Development
When to be surprised about validation data:When to be surprised about validation data:
Precision ofPrecision of
impurity determinationimpurity determination
Precision ofPrecision ofAPI determinationAPI determination
Method precision ofMethod precision of
released API (dissolution)released API (dissolution)
System precisionSystem precision % RSD 0.33 2.25% RSD 0.33 2.25
Method precisionMethod precision % RSD 0.0% RSD 0.0
Average peak areaAverage peak area % RSD 0.08% RSD 0.08
Acceptance criterionAcceptance criterion % RSD 2.0% RSD 2.0
Average peak areaAverage peak area % RSD 0.4% RSD 0.4
Acceptance criterionAcceptance criterion % RSD 10.0% RSD 10.0
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Birgit Schmauser | April 200848 |
Analytical Method DevelopmentAnalytical Method Development
Specification range (USL-LSL)Specification range (USL-LSL) Process variability (usuallyProcess variability (usually 2 SD) 2 SD)
Analytical variability (Analytical variability ( 3 3)) ~ NMT 30% of total specification range~ NMT 30% of total specification range
Analytical variabilityAnalytical variability Process variabilityProcess variability
Reliability of evaluation of major process variablesReliability of evaluation of major process variables by analyticalby analyticalproceduresprocedures depends ondepends on analytical variabilityanalytical variability
ImpuritiesImpurities
LOQLOQ and specification limit (e.g. qualification limits NMTand specification limit (e.g. qualification limits NMT0.15%)0.15%)
Response factorsResponse factors (LOQ modified by response factor)(LOQ modified by response factor)
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Birgit Schmauser | April 200849 |
Analytical Method DevelopmentAnalytical Method Development
Methods for cleaning validationMethods for cleaning validation Method for assay and related substances used in stability studies of APIMethod for assay and related substances used in stability studies of API
and FPPand FPP
SpecificitySpecificity (in(in samples taken from a cleaning assessmentsamples taken from a cleaning assessment))
LLinearity of responseinearity of response (from(from 50% of the cleaning limit to50% of the cleaning limit to 110x this0x this
concentrationconcentration; R; R
22
0.9900)0.9900) PrecisionPrecision
RepeatabilityRepeatability (RSD(RSD 5%)5%)
iinterntermediate precision [ruggedness (USP)]mediate precision [ruggedness (USP)]
ReproducibilityReproducibility
LLimits of detection and quantitationimits of detection and quantitation
AAccuracy or recovery from rinsateccuracy or recovery from rinsate (( 80%)80%), swabs, swabs (( 90%)90%), and, andprocess surfaceprocess surface (( 70%)70%)
RRangeange ((lowest level is at least 2x higher thanlowest level is at least 2x higher than LOQLOQ))
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Birgit Schmauser | April 200850 |
Analytical Method DevelopmentAnalytical Method Development
SummarySummary Analytical procedures play a critical role in pharmaceuticalAnalytical procedures play a critical role in pharmaceutical
equivalence and risk assessment/managementequivalence and risk assessment/management
Establishment ofEstablishment ofproduct-specificproduct-specific acceptance criteriaacceptance criteria
Assessment ofAssessment ofstabilitystability of APIs and FPPsof APIs and FPPs
Validation of analytical procedures should demonstrate thatValidation of analytical procedures should demonstrate that
they arethey are suitable for their intended usesuitable for their intended use
Validation of analytical procedures deservesValidation of analytical procedures deserves special attentionspecial attentionduring assessmentduring assessment of dossiersof dossiers for prequalificationfor prequalification
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Analytical Method DevelopmentAnalytical Method Development
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