Title:AlternativeAnalyticalMethodValidationinPharmaceuticals:Replacinga RegulatoryAnalyticalMethodinCleaningValidation Authors:StephenLawson,WillMcHaleandBrianWallace Thisretitledposterisreplacing: AlternativeAnalyticalMethodValidationinPharmaceuticals:RealWorldExamplesof WhySomeRegulatoryAnalyticalMethodsShouldbeReplaced

Introduction: Resultsobtainedfromanalyticalinstrumentationinthepharmaceuticalindustryundergo intensescrutinybecausetheyarequiteoftenusedtomakedecisionsthatultimately ensurethehealthandsafetyofconsumers.Whetheritbedatafromresearchand development,manufacturingorqualitycontrol,theaccuracyandprecisionoflaboratory andprocessanalyticalinstrumentationisofparamountimportanceinvalidatingdatafor useinalllevelsofdecisionmakingwithinapharmaceuticalfacility.Thescientificdetails ofinstrumentvalidationforthepharmaceuticalindustryareavailableinguidance documentsonthetopicofanalyticalproceduresandmethodsvalidation.Thisdiscussion focusesonacriticalsubsetofthetopicreferredtoasregulatoryandalternativeanalytical procedurerelationships,specificallyasitappliestotheanalyticalinstrumentationusedin 1 cleaningvalidation.2 RegulatoryandAlternativeAnalyticalProceduresaredefinedasfollows :

RegulatoryAnalyticalProcedureTheanalyticalprocedurebeingusedtoevaluatea definedcharacteristicofadrugsubstanceordrugproduct. AlternativeAnalyticalProcedureAnanalyticalprocedureproposedbyanapplicantfor useinplaceoftheregulatoryanalyticalprocedure.

Background: Itwastheinfamous1988recallofCholestyramineResinUSPthatsetintomotionthe rigorouslyscientificapproachtocleaningvalidationandverificationthepharmaceutical industryemploystoday.ReflectingbacktotheintroductionoftheFoodandDrug Administrations(FDAs)GuidetoInspectionsValidationofCleaningProcesses,a referencedocumentforFDApersonnel,itisclearthataparadigmshiftwastakingplace withrespecttothescienceofcleaningvalidation.Standardpracticeintheindustrytoday istheestablishmentofacceptableresiduelimits(orcarryoverlimits)foralldrugs manufacturedinafacility.Todaytheseresiduelimitsareverythoughtfullyderivedfrom thetoxicologyandefficacyknowledgegainedduringdrugdevelopmentsandclinical trials,butitwasnotsolongagothatcleaningverificationsinvolvedlittlemorethan organolepticinspectionsofmanufacturingequipment,withnomonitoringofcleaning programstomeetstrictresiduelimitcriteria.Alongwiththeemergenceofthese scientificallybasedlimitscameaneedtobotheffectivelysampleprocessequipmentand alsoaccuratelyobtainresultsfromanalyticalinstrumentationsothataccurate

determinationsofcleanlinesscanbemade.LiquidChromatography(LC)wasoneofthe firsttechniquesemployedincleaningvalidationsampleanalysisnotsimplybecauseofits qualitativeandquantitativecapabilities,butevenmorelikelybecauseofitswidespread 3,4 usethroughouttheindustry.

Discussion: TheuseofLCfortheanalysisofcleaningvalidation/verificationsamplesremainsthe mostcommoneventoday,butthattruthstemsmainlyfromitspracticalimplementation intotheapplicationsometwentyyearsago.LCsversatility,andthesound understandingthatindustryandagencyscientistssharedofitstheoryandoperationmade italogicalchoice. Ascleaningvalidationknowledgeandexperiencehasincreasedinrecentyears,new technologiesforsampleanalysishaveemergedthatofferimprovedperformance characteristicsoverHPLC.TotalOrganicCarbon(TOC)analysisisoneanalytical methodthathassignificantadvantagesascomparedtoHPLC,anditisbeingemployedin cleaningvalidationapplicationsinincreasingnumbersoflate.Thedetectionlimitsof TOC,itsaccuracyandprecisioncapabilitiesandmostimportantlyitsnonspecificnature makeitamuchmorepowerfultoolthanHPLCforcleaningvalidation.Nonspecificity, althoughquestionedbysomeasappropriate,allowsTOCuserstoscreenfor contaminationoutsideactiveingredientsanddetergentswitheveryanalysissomething chromatographictechniquesoftenfailtooffer.Evenwiththisknowledge,changehas beenslowtotakeplaceforanumberofreasons,nottheleastofwhichhasbeenthe absenceofasimplemigrationpathforthosefacilitiesalreadyhavingvalidatedHPLCfor 5,6,7 useintheircleaningvalidation/verification. TheFDAstatesthatthereareessentiallyfourcriteriayoumustconsidercarefullybefore 8 implementingananalyticalprocedureforacleaningvalidationapplication.Theyare: Sensitivity Itisnecessarytodemonstratethatthemethodisappropriateforthe establishedcontaminationlimitswithrespecttosensitivityandlimitsofdetection. Practicality Themethodshouldbepracticalandrapid,andwheneverpossible,itis desirabletoutilizepreviouslyexistingequipment(thatiswellknown/understoodbothby theuserandtheFDA). ValidationSchemeThemethodshallbesuchthatitcanbereadilyvalidatedin accordancewithregulatoryrequirements(FDA,ICH,etc.)forinstrumentation. SuccessfulRecoveryStudy Theanalyticaldevelopmentshallincludecompound recoverystudiesthatchallengethesamplingandtestingmethods. Itistypicallyacceptedthatthefollowingdetailsbeincludedinthedescriptionof 3 analyticalproceduresusedinpharmaceuticalmanufacturing : PrincipleAdescriptionofthebasicprinciplesoftheanalyticalprocedureshouldbe givenhere(separation,detection,etc.). SamplingJustificationforselectionofsampletypes,numbersandanalysiscriteria.

ReagentsDetailedlistofreagents,theirpreparations,associatedhazardsanddirections foruse. SystemSuitabilityTestingEnsuringaproperlyfunctioninganalyticalsystem, suitabilitystandardtestingparametersandassociatedacceptancecriteriashouldbe discussed.Confirmsthesystemwillproducereliableresultsindependentof environmentalconditions. PreparationofStandardsProceduresforthepreparationanduseofallstandard solutions(calibration,internal,stock,verification,working,etc.). SamplePreparation Samplepreparationsforindividualsampletestsshouldbedetailed. ProceduresFulldescriptionoftheanalyticalprocedureshouldbeprovided. CalculationsRepresentativecalculations,includingmathematicaltransformationsor formulasincludedindataanalysisandreportingmustbeincluded. ReportingofResultsCompleteformattingdetailsforreport,including,butnotlimited toretentiontimes,detectionlimitsandquantitationlimits.Mayincludeinformationfor bothdrugproductsandimpurities.

TOCMethodValidation TheTOCmethodvalidationsectionofyourdocumentationshouldincludethefollowing 2 validationcharacteristicsforthemethodyouhavedeveloped : Accuracy Themeasureofexactnessofananalyticalmethod,ortheclosenessof agreementbetweenthevaluewhichisacceptedeitherasaconventional,true valueoranacceptedreferencevalueandthevaluefound. Precision Precisionisthemeasureofthedegreeofrepeatabilityofananalytical methodundernormaloperationandisnormallyexpressedasthepercentrelative standarddeviationforastatisticallysignificantnumberofsamples. DetectionLimitThepointatwhichinstrumentresponseforananalyteor compound(carbonconcentration)canbedistinguishedfrominstrumentnoise,but notbeaccuratelyquantitated. QuantitationLimitThelowestconcentrationofcarbonthatcanbedetermined withacceptableprecisionandaccuracyunderthestatedoperationalconditionsof themethod. Linearity Linearityshouldbedemonstratedbyanrsquaredvalueindicatingthat theregressionlinewillbeanexcellentpredictorwhentransformingsampledata. RangeDetailsoftheinstrumentsanalyticalrange,includingrelevantmethod modificationsnecessarytoachieve. RobustnessAbilityoftheanalyticalproceduretoremainunaffectedbysmall butdeliberatevariationsinmethodparameters.

ThesensitivityofsomeTOCinstrumentsmakesthemaperfectfitforcleaningvalidation analysis.DatafromTable1indicatesthatthesensitivityofthetechniqueismorethan adequatetoqualifyitforuseincleaningvalidationapplications.Thesedetectionlimits

arearguablymoreimpressivethanchromatographicmethodswhenyouconsiderthe breadthofanalysisasfaraspotentialcontaminationtypes.Sample PurifiedWater Result (ppmCn=9) 0.0021 Std.Dev. (+/ppmC) 0.0006 MDL (ppmC) 0.0017 LOQ (ppmC) 0.0060

Table1 DataobtainedusingTekmarFusionTOCAnalyzer.

LinearityoverarangeofTOCvaluesappropriateforestablishedlimitsshouldbe demonstrated.Withcoefficientsofcorrelations>0.995generallydeemedacceptable, 2 TOCanalyzerscanachieveacceptablelinearityoverratherwideranges.Ther valueof 0.9996forTOCconcentrationsfrom020ppmshowninGraph1supportexcellent linearity.

Graph1 Linearitydatafrom020ppmTOCobtainedusingTekmarFusionTOCAnalyzer.

Concentration (ppmC)0.005 0.01 0.013 0.02 0.025 0.04 0.05 0.067 0.1 0.2 0.25 0.333 0.5

Result (ppmCn=3)0.0046(PASS) 0.0095(PASS) 0.0126(PASS) 0.0172(PASS) 0.0224(PASS) 0.0349(PASS) 0.0468(PASS) 0.0608(PASS) 0.0939(PASS) 0.1957(PASS) 0.2439(PASS) 0.3294(PASS) 0.5074(PASS)

Std.Dev. (+/ppmC)0.0017 0.0021 0.0004 0.0008 0.002 0.0013 0.003 0.0026 0.0013 0.0024 0.0021 0.0038 0.0024

RSDN/A N/A 2.88% 4.90% 9.00% 3.64% 6.44% 4.22% 1.39% 1.25% 0.87% 1.16% 0.47%

Dilution1:200 1:100 1:75 1:50 1:40 1:25 1:20 1:15 1:10 1:5 1:4 1:3 1:2

Min/MaxCriteria(ppmC) (85115%dev).0043/.0058 .0085/.0115 .0113/.0153 .0170/.0230 .0213/.0288 .0340/.0460 .0425/.0575 .0567/.0767 .0850/.1150 .1700/.2300 .2125/.2875 .2833/.3833 .4250/.5750

Table2 AutoCheckStandardDatadilutedfrom1ppmCKHPTOCStockStandardfor13points(n=3replicatesper standard)automaticallyusingtheTekmarFusionTOCAnalyzer.

Selectionofrecoverydataforcompoundsusedinmanufacturingbyseveral manufacturersshowsthattheobligationtodemonstratesuccessfulproductrecoveriesis met,withvaluesrangingfrom89%108%(Table2).FDA,WHOandgeneralindustry 4,9 criteriarangefrom50%80%forwhatareconsideredacceptablelevels.Molecular Weight14.50kDa

CompoundBovineIgG

Carbon% (eestimate)51e

Carbonin Standard(ppm)1.0

Measured Carbonby TOC(ppm)1.11

Percent Recovery (%)108

TrypsinInhibitor

21.50kDa

49e

1.0

0.87

89

LGlutamicAcid

147.13g/mol

40.81

1.0

1.04

104

LTryptophan CitricAcid

204.23 192.13

64.69 37.50

1.0 1.0

1.09 1.03

109 103

Table3 DataobtainedusingPhoenix8000TOCAnalyzer.

Conclusion: TheadvantagesofTOCforcleaningvalidationusearemany,yetmostlabsfind themselvesinthepositionofalreadyhavingavalidatedregulatoryanalyticalprocedure inplace.DefendingachangetoanalternativeanalyticalprocedurelikeTOCdoesnt needtobeadauntingtaskandshouldrequirenomorethantwoweeksasfarasdata

collection.Followingtherequirementsdetailedinregulatoryguidelinesforanalytical instrumentqualificationandmethodvalidationisallthatisnecessarytobeableto confidentlydefendyourdecisiontochange.Rememberthatoneofthemostimportant attributesofTOCthatshouldbedetailedinadiscussionofanalysisprinciplesisitsnon specificnatureandinherentabilitytodetectandquantifycontaminationthat chromatographicmethodsmaynot. References: 1. USFDA.CurrentGoodManufacturingPracticeRegulations,21CFR211.67. 2. USDHHS,CDER(RadhikaRajagopalan)andCBER(AlfredDelGrosso). AnalyticalProceduresandMethodsValidation,DraftGuidanceforChemistry, ManufacturingandControls,August2000. 3. G.Bismuth,CleaningValidation:APracticalApproach,CRCPress,2000,p24. 4. USFDA.GuidetoInspectionsofValidationofCleaningProcedures,July1993. 5. J.G.JinandCherylWoodward,DevelopmentofTotalOrganicCarbonAnalysis forDetergentResidueVerification,J.ValidationTechnology,Vol.7No.4, 2001,pp.278 284. 6. K.M.Jenkinsetal., ApplicationofTotalOrganicCarbonAnalysistoCleaning Validation,PDAJ.Pham.Sci.&Tech.,Vol.50No.1,1996,pp.615. 7. H.J.Kaiser,MeasurementofOrganicandInorganicResiduesRecoveredfrom Surfaces,J.ValidationTechnology,Vol.6No.1,1999,pp.424 436. RCPress, Jan2000. 8. Bansal,SurendraK.,etal,QualificationofAnalyticalInstrumentsforUseinthe PharmaceuticalIndustry:AScientificApproach,AAPSPharmSciTech20045 (1)Article22. 9. WHOGuidetoGoodManufacturingPractices,Part2 Validation, WHO/VSQ/97.02.