HANDBOOKOFGENERICDRUGDEVELOPMENTinfo@iagim.orgANDADEVELOPMENTEdNumber:01EffectiveDateAPPROVEDEd.Status:NewDD/MM/YY_______________________________________________/________DepartmentR&DRAQC/QA24VOLUMEDRUGDEVELOPMENTSERIES:Sect:15.5ANDAANDADevelopmentSECTIONXVSECTION15AnalyticalMethods(TYPICALANALYTICALMETHODVALIDATION)1.PURPOSEThepurposeofthisStandardAnalyticalProcedureistodemonstratetheprocedurerequiredtovalidatein-houseHPLCanalyticalmethodsandtoshowthatthemethodsarestability-indicating.MethodsbasedontheUSPbutmodifiedforstabilityindicatingtestpurposesrequirefullin-housevalidation.ThisprocedureensuresthattheProductDevelopmentProcessandProcessQualificationBatchanalysisisbasedonafoundationofGoodLaboratoryPracticeusingvalidatedtestprocedures.2.RESPONSIBILITYTheHeadofAnalyticalDevelopmentincoordinationwiththemanagersofQCandRegulatoryAffairsattheproposedmanufacturingsite.3.FREQUENCYForeachnon-compendialanalyticalmethodintendedforANDA(orOTCANDA)manufacturedproducts.ForStability-IndicatingAssaysandlimittestingofimpuritiesthatmaybebasedoncompendialmethods.EachProductstrengthwillfollowthefullmethodvalidationprocedure.4.PROCEDURE[a].MethodValidationNon-compendialmethodsvalidationwillfollowtheUSPdirectionforparametersneededforthevalidationoftestmethods.Typicalparametersforvalidatingassaysandothernon-compendialanalyticalmethodsdesignedforprovidingquantitativeresultsshallinclude:•Accuracy•Recovery•Precision(Systemreproducibility,Methodreproducibility)•Specificity•Linearity•Range•Ruggedness(differentanalysts/days/differentequipmentmodels/columns)[b].PlaceboAnalysis.Amixtureofnon-actives(placebo)shallbepreparedandsubjectedtoanalysis.Nointerferingpeaksshallbeobservedinthegraphoftheplacebochromatogram.HANDBOOKOFGENERICDRUGDEVELOPMENTinfo@iagim.orgANDADEVELOPMENTEdNumber:01EffectiveDateAPPROVEDEd.Status:NewDD/MM/YY_______________________________________________/________DepartmentR&DRAQC/QA24VOLUMEDRUGDEVELOPMENTSERIES:Sect:15.6ANDAANDADevelopment[c].ThestabilityoftheStandardsolutionisassessedbyre-injectionofthestandardsolutionafter24xnhours(wheren=numberofdaystheStandardwillbeused).StandardPreparationforAssayComparisonofstandardsolutionsforAssayofActivematerial,injectedafteronemonthandfreshlyprepareddemonstratethatthestandardsolutionsarestableanddoesnotloseitsqualityafteronemonthifrefrigerated.StandardPreparationforImpurityComparisonofstandardsolutionsofGuanine,injectedafteronemonthandfreshlyprepareddemonstratethatthestandardsolutionsarestableanddoesnotloseitsqualityafter1monthifrefrigerated.NameofstandardsStorageconditionsDifference.relativetofreshlypreparedstandard[Active]100%4°C2%[Impurity]100%4°C2%StandardSolutionsarestoredatcontrolledtemperaturesandlightconditionsasperlabeling.[d].StabilityIndicatingProcedures.FortheStabilityIndicatingMethod,theproductsampleshallincludeforceddegradationbystressedanalysis.Conditionsofconcentrationandreactiontimemayvarydependingontheactivedrugsubstanceanddrugproducte.g.:•Oxidation-(H2O2plusstandingtime).•BaseHydrolysis-(NaOHxNplusstandingtime).•AcidHydrolysis-(HClconc.plusstandingtime).•Sunlight-(24hoursstandingtime).•Heat-(xdegreesC).SummaryofStabilityIndicatingResultsStressedConditionsTemp.TimeRawMaterial;Tablets(°°C)(hr)RemainingSubstance.(%)PeakPurity,(Figure)RemainingSubstance(%)PeakPurity,(Figure)Solutionheating9012100.2pure98pureSolidheating1602101.3pure92pureSunlight765w/m24014101.1pure84.8pure3,3NSodiumHydroxide701099.8pure100.2pure10%HydrogenPeroxide37377.5pure90.5pure5%HydrochloricAcidRoom2079.7pure78.6pureHANDBOOKOFGENERICDRUGDEVELOPMENTinfo@iagim.orgANDADEVELOPMENTEdNumber:01EffectiveDateAPPROVEDEd.Status:NewDD/MM/YY_______________________________________________/________DepartmentR&DRAQC/QA24VOLUMEDRUGDEVELOPMENTSERIES:Sect:15.7ANDAANDADevelopment[e]SpecificityandSuitability(ResolutionandTailingFactors).Whenasatisfactoryseparationsofallthedegradationpeakshavebeenachievedthroughtheforceddegradationreactions,aResolutionFactor(accordingtotheUSPrequirements)betweenthemainactivepeakandthenearestdegradantpeakiscalculatedusingtheUSPformula.ATailingFactor(accordingtotheUSPformula)iscalculatedforthemainactivepeak.[f]SystemSuitabilityTestAmixtureof[Active]AS.standardattheconcentrationabout[0.1]mg/mLandof[Impurity]AS.standardattheconcentrationabout[0.01]mg/mLaccordingtoMethodSI-1000waspreparedandinjectedintotheHPLCsystem.Forchromatogramobtainedthefollowingvalueswerecalculated(accordingtoUSP):1.RelativeRetentionTimefor[Impurity]peakRRT=RT[Impurity]=2.65=0.31RT[Active]8.452.Tailingfactorfor[Active]peakT=W2=94.2=1.1f0.05fThevaluesdepictthespecificityofthemethodforresolutionbetweenthemainpeakandimpuritypeak.(valuesshownfordemonstrationspurposes).PeakPurityThephotodiode-arrayisusedfortheevaluationofthestabilityindicatingnatureoftheassaymethodnumberSI-1000for[000]mgand[000]mgtabletsusingaWaters996™Unit,controlledbythechromatographymanagerMillennium2010™.Peakpurityandmatchresultsarereportedas:PurityAngleisameasureofspectralnon-homogeneityacrossapeak-i.e.theweighedaverageofallSpectralContrastA