Analytical method validation for FDA complianceFDA的依从性分析方法的验证

PHARM46212009EuropeanandInternationalregulatorybodiesandtheirguidelinesondifferentaspectsofQABodyFullnameGuidanceonEurachemFocusforAnalyticalChemistryinEuropeMethodvalidationCITACCooperationofInternationalTraceabilityinAnalyticalChemistryProficiencytestingQualityAssuranceEAEuropeanCooperationforAccreditationAccreditationCENEuropeanCommitteeforNormalizationStandardizationIUPACInternationalUnionofPure&AppliedChem.MethodvalidationISOInternationalStandardizationOrganisationStandardisationAOACILACAssociationofOfficialAnalyticalChemistsInternationalLaboratoryAccreditationCooperat.Internalqual.ControlProficiencytestingAccreditationFDAUSFoodandDrugAdministrationMethodvalidationUSPUnitedStatesPharmacopoeiaMethodvalidationICHInternationalConferenceonHarmonizationMethodvalidation22009MethodValidationValidationofanalyticalproceduresistheprocessofdeterminingthesuitabilityofagivenmethodologyforprovidingusefulanalyticaldata.J.Guerra,Pharm.Tech.March1986Validationistheformalandsystematicproofthatamethodcompileswiththerequirementsfortestingaproductwhenobservingadefinedprocedures.G.Maldener,Chromatographia,July198932009Methodvalidationistheprocessofdemonstratingthatanalyticalproceduresaresuitablefortheirintendeduseandthattheysupporttheidentity,strength,quality,purityandpotencyofthedrugsubstancesanddrugproductsMethodvalidationisprimarilyconcernedwith:identificationofthesourcesofpotentialerrorsquantificationofthepotentialerrorsinthemethodAnmethodvalidationdescribesinmathematicalandquantifiabletermstheperformancecharacteristicsofanassay42009ExamplesofMethodsThatRequire

ValidationDocumentationChromatographicMethods-HPLC,GC,TLC,GC/MS,etc.PharmaceuticalAnalysis-InsupportofCMC.BioanalyticalAnalysis-InsupportofPK/PD/ClinicalStudies.SpectrophotometricMethods–UV/VIS,IR,NIR,AA,NMR,XRD,MSCapillaryElectrophoresisMethods-Zone,IsoelectricFocusingParticleSizeAnalysisMethods-Laser,Microscopic,Sieving,SEC,etc.AutomatedAnalyticalMethods-Robots,AutomatedAnalysis.52009ConsiderationsPriorto

MethodValidationSuitabilityofInstrumentStatusofQualificationandCalibrationSuitabilityofMaterialsStatusofReferenceStandards,Reagents,PlaceboLotsSuitabilityofAnalystStatusofTrainingandQualificationRecordsSuitabilityofDocumentationWrittenanalyticalprocedureandproperapprovedprotocolwithpre-establishedacceptancecriteria62009ValidationStep

Definetheapplication,purposeandscopeofthemethod.Analytes?Concentration?Samplematrices?Developaanalyticalmethod.Developavalidationprotocol.Qualificationofinstrument.Qualify/trainoperatorQualificationofmaterial.Performpre-validationexperiments.Adjustmethodparametersand/oracceptancecriteriaifnecessary.Performfullvalidationexperiments.DevelopSOPforexecutingthemethodinroutineanalysis.Documentvalidationexperimentsandresultsinthevalidationreport.72009PurposeofMethodValidationIdentificationofSourcesandQuantitationofPotentialerrorsDeterminationifMethodisAcceptableforIntendedUseEstablishProofthataMethodCanbeUsedforDecisionMakingSatisfyFDARequirements82009Whatisnot

AnalyticalMethodValidation?Calibration

TheProcessofPerformingTestsonIndividualSystemComponentstoEnsureProperfunctionForexample)HPLCDetectorcalibrationWavelengthAccuracy/LinearRange/NoiseLevel/Drift92009SystemSuitability

Testtoverifytheproperfunctioningoftheoperatingsystem,i.e.,theelectronics,theequipment,thespecimensandtheanalyticaloperations.MinimumResolutionof3.0betweentheanalytepeakandinternalstandardpeaksRelativeStandardDeviationofreplicatestandardinjectionsofnotmorethan2.0%10200911SystemSuitabilitySampleValidationMethodAnalystCalibrationPumpDetectorInjectorDataSystem2009MethodLifeCycle12ValidationDevelopmentOptimization2009Verificationvs.ValidationCompendialvs.Non-compendialMethodsCompendialmethods-VerificationNon-compendialmethods-Validationrequirement132009CompendialAnalyticalProceduresTheAnalyticalproceduresintheUSP25/NF20arelegallyrecognizedundersection501(b)oftheFederalFood,DrugandCosmeticActastheregulatoryanalyticalproceduresforthecompendialitems.Thesuitabilityoftheseproceduresmustbeverifiedunderactualconditionsofuse.WhenusingUSP25/NF20analyticalprocedures,theguidancerecommendsthatinformationbeprovidedforthefollowingcharacteristics:

SpecificityoftheprocedureStabilityofthesamplesolutionIntermediateprecision

142009PublishedValidationGuidelines1978CurrentGoodManufacturingPractices(cGMPs)1987FDAValidationGuideline1989Supplement9toUSPXXI1994CDERReviewerGuidance:ValidationofChromatographicMethod1995ICHValidationDefinitions:Q2A,TextonValidationofAnalyticalprocedures1997ICHValidationMethodology:Q2B,ValidationofAnalyticalProcedures:Methodology1999Supplement10toUSP23<1225>:ValidationofCompendialMethods1999CDER“BioanalyticalMethodValidationforHumanStudies”

2000CDERDraft“AnalyticalProceduresandMethodValidation”152009RegulatoryandCompliance

RequirementsReviewValidationofananalyticalmethodistheprocessbywhichitisestablished,bylaboratorystudies,thattheperformancecharacteristicsofthemethodmeettherequirementsfortheintendedanalyticalapplications16USP23GeneralInformation<1225>2009Theaccuracy,sensitivity,specificity,andreproducibilityoftestmethodsemployedbythefirmshallbeestablishedanddocumented.Suchvalidationanddocumentationmaybeaccomplishedinaccordancewith211.194(a)(2).1721CFRPART211-CURRENTGOODMANUFACTURINGPRACTICEFORFINISHEDPHARMACEUTICALSSubpartI-LaboratoryControls211.165Testingandreleasefordistribution(e)2009Theobjectiveofvalidationofananalyticalprocedureistodemonstratethatitissuitableforitsintendedpurpose18ICHGuidelineforIndustryQ2A,TextonValidationofAnalyticalProceduresMarch19952009Inpractice,itisusuallypossibletodesigntheexperimentalworksuchthattheappropriatevalidationcharacteristicscanbeconsideredsimultaneouslytoprovideasound,overallknowledgeofthecapabilitiesoftheanalyticalprocedure,forinstance:Specificity,Linearity,Range,Accuracy,and Precision.19ICHGuidelineforIndustryQ2B,ValidationofAnalyticalProcedures:Methodology2009Today’sValidationRequirements20ICH/USPGMPs(legal)FDA2009ICH/USPValidationRequirements&ParametersSpecificityLinearityRangeAccuracyPrecisionRepeatabilityIntermediatePrecisionReproducibilityLimitofDetectionLimitofQuantitation21ICHSpecificityLinearityandRangeAccuracyPrecisionLimitofDetectionLimitofQuantitationRuggednessRobustnessUSP2009USPDataElementsRequired

ForAssayValidation22AnalyticalPerformanceParameterAssayCategory1AssayCategory2AssayCategory3QuantitativeLimitTestsAccuracyYesYes**PrecisionYesYesNoYesSpecificityYesYesYes*LODNoNoYes*LOQNoYesNo*LinearityYesYesNo*RangeYesYes**RuggednessYesYesYesYes*Mayberequired,dependingonthenatureofthespecifictest.2009USPCategoriesCategory1:Quantitationofmajorcomponentsor activeingredientsCategory2:Determinationofimpuritiesor degradationproductsCategory3:Determinationofperformance characteristics232009ICHValidationCharacteristicsvs.

TypeofAnalyticalProcedure24TypeofAnalyticalProcedureIdentificationImpuritytestingAssayQuantitativeLimitTestsAccuracyNoYesNoYesPrecisionRepeatabilityNoYesNoYesInterm.Prec.NoYesNoYesSpecificityYesYesYesYesLODNoNoYesNoLOQNoYesNoNoLinearityNoYesNoYesRangeNoYesNoYes2009Specificity/SelectivityAbilityofananalyticalmethodtomeasuretheanalytefreefrominterferenceduetoothercomponents.SelectivitydescribestheabilityofananalyticalmethodtodifferentiatevarioussubstancesinasampleOriginaltermusedinUSPAlsoPreferredbyIUPACandAOACAlsousedtocharacterizechromatographiccolumnsDegreeofBias(UsedinUSP)Thedifferenceinassayresultsbetweenthetwogroupsthesamplecontainingaddedimpurities,degradationproducts,relatedchemicalcompounds,placeboingredientsthesamplewithoutaddedsubstances

252009Specificity:ImpuritiesAssayChromatographicMethodsDemonstrateResolutionImpurities/DegradantsAvailableSpikewithimpurities/degradantsShowresolutionandalackofinterferenceImpurities/DegradantsNotAvailableStressSamplesForassay,StressedandUnstressedSamplesshouldbecompared.Forimpuritytest,impurityprofilesshouldbecompared.262009ForcedDegradationStudiesTemperature(50-60℃)Humidity(70-80%)AcidHydrolysis(0.1NHCl)BaseHydrolysis(0.1NNaOH)Oxidation(3-30%)Light(UV/Vis/Fl)Intentistocreate10to30%Degradation272009LinearityAbilityofanassaytoelicitadirectandproportionalresponsetochangesinanalyteconcentration.282009LinearityShouldbeEvaluatedByVisualInspectionofplotofsignalsvs.analyteconcentrationByAppropriatestatisticalmethodsLinearRegression(y=mx+b)CorrelationCoefficient,y-intercept(b),slope(m)Acceptancecriteria:Linearregressionr2>0.95

Requiresaminimumof5concentrationlevels292009RangeAcceptablerangehavinglinearity,accuracy,precision.ForDrugSubstance&DrugproductAssay80to120%oftestConcentrationForContentUniformityAssay70to130%oftestConcentrationForDissolutionTestMethod+/-20%overentireSpecificationRangeForImpurityAssaysFromReportingLevelto120%ofImpuritySpecificationforImpurityAssaysFromReportingLevelto120%ofAssaySpecificationforImpurity/AssayMethods302009AccuracyClosenessofthetestresultsobtainedbythemethodtothetruevalue.31AccuracyShouldbeestablishedacrossspecifiedrangeofanalyticalprocedure.Shouldbeassessedusingaminimumof3concentrationlevels,eachintriplicate(totalof9determinations)Shouldbereportedas:PercentrecoveryofknownamountaddedorThedifferencebetweenthemeanassayresultandtheacceptedvalue322009AccuracyDataSet(1of3)33AmountAdded(mg)AmountFound(mg)PercentRecovery0.00.0---50.250.4100.579.680.1100.699.9100.7100.8120.2119.899.7150.4149.799.52009PrecisionTheclosenessofagreement(degreeofscatter)betweenaseriesofmeasurementsobtainedfrommultiplesamplingsofthesamehomogeneoussample.Shouldbeinvestigatedusinghomogeneous,authenticsamples.342009Precision…Consideredat3LevelsRepeatabilityIntermediatePrecisionReproducibility352009RepeatabilityExpresstheprecisionunderthesameoperatingconditionsoverashortintervaloftime.AlsoreferredtoasIntra-assayprecision36Shouldbeassessedusingminimumof9determinations(3concentrations/3replicates)orMinimumof6determinationsatthe100%level.2009IntermediatePrecision37Expresswithin-laboratoryvariations.Expressedintermsofstandarddeviation,relativestandarddeviation(coefficientofvariation)andconfidenceinterval.Dependsonthecircumstancesunderwhichtheprocedureisintendedtobeused.Studiesshouldincludevaryingdays,analysts,equipment,etc.2009Repeatability&IntermediatePrecisionDay1Day2100.699.5100.899.9100.198.9100.399.2100.599.7100.499.638GrandMean=100.0RSD=0.59%Mean=100.5RSD=0.24%Mean=99.5RSD=0.36%2009ReproducibilityDefinition:AbilityreproducedatawithinthepredefinedprecisionDetermination:SD,RSDandconfidenceintervalRepeatabilitytestattwodifferentlabs.Note:DatanotrequiredforBLA/NDALab1Lab2Lab3Day1Day2Day1Day2Day1Day2Man1Man2Man1Man2Man1Man23Prep3Prep3Prep3Prep3Prep3Prep39DetectionLimit(LOD)/QuantitationLimit(LOQ)LODLowestamountofanalyteinasamplethatcanbedetectedbutnotnecessarilyquantitated.EstimatedbySignaltoNoiseRatioof3:1.40LOQLowestamountofanalyteinasamplethatcanbequantifiedwithsuitableaccuracyandprecision.EstimatedbySignaltoNoiseRatioof10:1.2009BasedinVisualEvaluations-Usedfornon-instrumentalmethodsBasedonSignal-toNoise-Ratio-3:1forDetectionLimit-10:1forQuantitationLimitBasedonStandardDeviationoftheResponseandtheSlope41LODandLOQEstimatedby2009S=slopeofcalibrationcurves=standarddeviationofblankreadingsor

standarddeviationofregressionlineValidatedbyassayingsamplesatDLorQL42DL=3.3sQL=10sSSLODandLOQEstimatedby200943YblLODLOQStatisticalestimateofLOD&LOQLOD=3.3Sbl/bLOQ=10Sbl/bY=bX+a2009Definition:CapacitytoremainunaffectedbysmallbutdeliberatevariationsinmethodparametersDetermination:ComparisonresultsunderdifferingconditionswithprecisionundernormalconditionsExamplesoftypicalvariationsinLCInfluenceofvariationsofpHinamobilephaseInfluenceofvariationsinmobilephasecompositionDifferentcolumns(differentlotsand/orsuppliers)TemperatureFlowrate44Robustness2009RuggednessDegreeofreproducibilityoftestresultsunderavarietyofconditionsDifferentLaboratoriesDifferentAnalystsDifferentInstrumentsDifferentReagentsDifferentDaysEtc.Expressedas%RSD452009ICH/USPSystemSuitabilityICHDefinition:evaluationofequipment,electronic,analyticaloperationsandsamplesasawholeDetermination:repeatability,tailingfactor(T),capacityfactor(k’),resolution(R),andtheoreticalPlates(N)462009USP23<621>SystemSuitabilityRequirements47Parameters

RecommendationsK’Ingeneralk’≥2.0RR>2,betweenthepeakofinterestandtheclosestpotentialinterferent(degradant,internalSTD,impurity,excipient,etc…..)TT≤2NIngeneralN>2000RepeatabilityRSD≤2.0%(n≥5)2009Re-validationWhenMethodparametershavebeenchangedThescopeofthemethodhasbeenchangedSyntheticmethodshavebeenchangedImpurityprofilehasbeenchangedWhatPreferablyeverything.Exceptionsshouldbescientificallyjustified482009HowdoweKnowtheexpectationsoftheFDA?FDAForm483FDAWarningLettersPersonalExperiences492009483ObservationsTherewasinadequatemethodvalidationspecificitydatatodemonstratethateachmethodwascapableofdistinguishingtheactiveingredientfromitsimpuritiesanddegradationproducts.Specificitystudiesdidnotincludetheminimumstressconditionsofacidandbasehydrolysis,oxidation,thermaldegradationandphotolysis,degradationschematicfortheactive