FDA微生物化验室的检查指南英文稿电子版本

1、Good is good, but better carries it.精益求精，善益求善。FDA微生物化验室的检查指南英文稿-GUIDETOINSPECTIONSOFMICROBIOLOGICALPHARMACEUTICALQUALITYCONTROLLABORATORIESNote:ThisdocumentisreferencematerialforinvestigatorsandotherFDApersonnel.ThedocumentdoesnotbindFDA,anddoesnotconferanyrights,privileges,benefits,orimmunitiesforo

2、ronanyperson(s).I.INTRODUCTIONTheGuidetotheInspectionofPharmaceuticalQualityControlLaboratoriesprovidedverylimitedguidanceonthematterofinspectionofmicrobiologicallaboratories.Whilethatguideaddressesmanyoftheissuesassociatedwiththechemicalaspectoflaboratoryanalysisofpharmaceuticals,thisdocumentwillse

3、rveasaguidetotheinspectionofthemicrobiologyanalyticalprocess.Aswithanylaboratoryinspection,itisrecommendedthatananalyst(microbiologist)whoisfamiliarwiththetestsbeinginspectedparticipateintheseinspections.II.MICROBIOLOGICALTESTINGOFNON-STERILEPRODUCTSForavarietyofreasons,wehaveseenanumberofproblemsas

4、sociatedwiththemicrobiologicalcontaminationoftopicaldrugproducts,nasalsolutionsandinhalationproducts.TheUSPMicrobiologicalAttributesChapterprovideslittlespecificguidanceotherthanThesignificanceofmicroorganismsinnon-sterilepharmaceuticalproductsshouldbeevaluatedintermsoftheuseoftheproduct,thenatureof

5、theproduct,andthepotentialhazardtotheuser.TheUSPrecommendsthatcertaincategoriesberoutinelytestedfortotalcountsandspecifiedindicatormicrobialcontaminants.Forexamplenaturalplant,animalandsomemineralproductsforSalmonella,oralliquidsforE.Coli,topicalsforP.aeruginosaandS.Aureus,andarticlesintendedforrect

6、al,urethral,orvaginaladministrationforyeastsandmolds.Anumberofspecificmonographsalsoincludedefinitivemicrobiallimits.Asageneralguideforacceptablelevelsandtypesofmicrobiologicalcontaminationinproducts,Dr.DunniganoftheBureauofMedicineoftheFDAcommentedonthehealthhazard.In1970,hesaidthattopicalpreparati

7、onscontaminatedwithgramnegativeorganismsareaprobablemoderatetoserioushealthhazard.Throughtheliteratureandthroughourinvestigations,ithasbeenshownthatavarietyofinfectionshavebeentracedtothegramnegativecontaminationoftopicalproducts.TheclassicalexamplebeingthePseudomonascepaciacontaminationofPovidoneIo

8、dineproductsreportedbyahospitalinMassachusettsseveralyearsago.Therefore,eachcompanyisexpectedtodevelopmicrobialspecificationsfortheirnon-sterileproducts.Likewise,theUSPMicrobialLimitsChapterprovidesmethodologyforselectedindicatororganisms,butnotallobjectionableorganisms.Forexample,itiswidelyrecogniz

9、edthatPseudomonascepaciaisobjectionableiffoundinatopicalproductornasalsolutioninhighnumbers;yet,therearenotestmethodsprovidedintheUSPthatwillenabletheidentificationofthepresenceofthismicroorganism.ArelevantexampleofthisproblemistherecallofMetaproterenolSulfateInhalationSolution.TheUSPXXIImonographre

10、quiresnomicrobialtestingforthisproduct.TheagencyclassifiedthisasaClassIrecallbecausetheproductwascontaminatedwithPseudomonasgladioli/cepacia.Thehealthhazardevaluationcommentedthattheriskofpulmonaryinfectionisespeciallyseriousandpotentiallylife-threateningtopatientswithchronicobstructiveairwaydisease

11、,cysticfibrosis,andimmuno-compromisedpatients.Additionally,theseorganismswouldnothavebeenidentifiedbytestingproceduresdelineatedinthegeneralMicrobialLimitssectionoftheCompendia.TheUSPcurrentlyprovidesforretestsintheMicrobialLimitssectionhoweverthereisacurrentproposaltoremovetheretestprovision.Aswith

12、anyothertest,theresultsofinitialtestshouldbereviewedandinvestigated.Microbiologicalcontaminationisnotevenlydispersedthroughoutalotorsampleofproductandfindingacontaminantinonesampleandnotinanotherdoesnotdiscountthefindingsoftheinitialsampleresults.Retestresultsshouldbereviewedandevaluated,andparticul

13、aremphasisshouldbeplacedonthelogicandrationaleforconductingtheretest.Inordertoisolatespecificmicrobialcontaminants,FDAlaboratories,aswellasmanyintheindustry,employsometypeofenrichmentmediacontaininginactivators,suchasTweenorlecithin.Thisisessentialtoinactivatepreservativesusuallypresentinthesetypeso

14、fproductandprovidesabettermediumfordamagedorslowgrowingcells.Othergrowthparametersincludealowertemperatureandlongerincubationtime(atleast5days)thatprovideabettersurvivalconditionfordamagedorslow-growingcells.Forexample,FDAlaboratoriesusethetestproceduresforcosmeticsintheBacteriologicalAnalyticalManu

15、al(BAM),6thEdition,toidentifycontaminationinnon-steriledrugproducts.Thistestingincludesanenrichmentofasampleinmodifiedletheenbroth.Afterincubation,furtheridentificationiscarriedoutonBloodAgarPlatesandMacConkeyAgarPlates.Isolatedcoloniesarethenidentified.ThisprocedureallowsFDAmicrobiologiststooptimiz

16、etherecoveryofallpotentialpathogensandtoquantitateandspeciateallrecoveredorganisms.AnotherimportantaspectofproceduresusedbyFDAanalystsistodeterminegrowthpromotioncharacteristicsforallofthemediaused.Theselectionoftheappropriateneutralizingagentsarelargelydependentuponthepreservativeandformulationofth

17、eproductunderevaluation.Ifthereisgrowthintheenrichmentbroth,transfertomoreselectiveagarmediaorsuitableenrichmentagarmaybenecessaryforsubsequentidentification.MicrobiologicaltestingmayincludeanidentificationofcoloniesfoundduringtheTotalAerobicPlateCounttest.Again,theidentificationshouldnotmerelybelim

18、itedtotheUSPindicatororganisms.TheimportanceofidentifyingallisolatesfromeitherorbothTotalPlateCounttestingandenrichmenttestingwilldependupontheproductanditsintendeduse.Obviously,ifanoralsoliddosageformsuchasatabletistested,itmaybeacceptabletoidentifyisolateswhentestingshowshighlevels.However,forothe

19、rproductssuchastopicals,inhalantsornasalsolutionswherethereisamajorconcernformicrobiologicalcontamination,isolatesfromplatecounts,aswellasenrichmenttesting,shouldbeidentified.III.FACILITIES,EQUIPMENT,ANDMEDIABegintheinspectionwithareviewofanalysesbeingconductedandinspecttheplatesandtubesofmediabeing

20、incubated(cautionshouldbeexercisednottoinadvertentlycontaminateplatesortubesofmediaontest).Beparticularlyalertforreteststhathavenotbeendocumentedand“specialprojects”inwhichinvestigationsofcontaminationproblemshavebeenidentified.Thiscanbeevaluatedbyreviewingtheongoinganalyses(productorenvironmental)f

21、orpositivetestresults.Requesttoreviewthepreviousdaysplatesandmedia,ifavailableandcompareyourobservationstotherecordedentriesinthelogs.Inspecttheautoclavesusedforthesterilizationofmedia.Autoclavesmaylacktheabilitytodisplacesteamwithsterilefilteredair.Forsealedbottlesofmedia,thiswouldnotpresentaproble

22、m.However,fornon-sealedbottlesorflasksofmedia,non-sterileairhasledtothecontaminationofmedia.Inaddition,autoclavinglessthantherequiredtimewillalsoallowmediaassociatedcontaminantstogrowandcauseafalsepositiveresult.Theseproblemsmaybemoreprevalentinlaboratorieswithaheavyworkload.Checkthetemperatureofthe

23、autoclavesinceoverheatingcandenatureandevencharnecessarynutrients.Thisallowsforalessthanoptimalrecoveryofalreadystressedmicroorganisms.Theobviousproblemwithpotentialfalsepositivesistheinabilitytodifferentiatebetweeninadvertentmediumcontaminationandtruecontaminationdirectlyassociatedwiththesampletest

24、ed.IV.STERILITYTESTINGOn10/11/91,theAgencypublishedaproposedruleregardingthemanufactureofdrugproductsbyasepticprocessingandterminalsterilization.Alistofcontaminatedorpotentiallycontaminateddrugproductsmadebyasepticprocessingandlaterrecalledwasalsomadeavailable.Manyoftheinvestigations/inspectionsofth

25、erecalledproductsstartedwithalistofinitialsterilitytestfailures.FDAreviewofthemanufacturersproduction,controls,investigationsandtheirinadequacies,coupledwiththeevidenceofproductfailure(initialsterilitytestfailure)ultimatelyledtotheaction.TheUSPpointsoutthatthefacilitiesusedtoconductsterilitytestssho

26、uldbesimilartothoseusedformanufacturingproduct.TheUSPstates,Thefacilityforsterilitytestingshouldbesuchastooffernogreateramicrobialchallengetothearticlesbeingtestedthanthatofanasepticprocessingproductionfacility.Properdesignwould,therefore,includeagowningareaandpass-throughairlock.Environmentalmonito

27、ringandgowningshouldbeequivalenttothatusedformanufacturingproduct.Sinceanumberofproductandmediamanipulationsareinvolvedinconductingasterilitytest,itisrecommendedthattheinspectionincludeactualobservationofthesterilitytesteventhoughsomecompanieshavetriedtodiscourageinspectiononthegroundsthatitmaymaket

28、hefirmsanalystnervous.Theinspectionteamisexpectedtobesensitivetothisconcernandmaketheobservationsinamannerthatwillcreatetheleastamountofdisruptioninthenormaloperatingenvironment.Nevertheless,suchconcernsarenotsufficientcauseforyoutosuspendthisportionoftheinspection.Oneofthemostimportantaspectsofthei

29、nspectionofasterilityanalyticalprogramistoreviewrecordsofinitialpositivesterilitytestresults.Requestlistsoftestfailurestofacilitatereviewofproductionandcontrolrecordsandinvestigationreports.Particularly,forthehighriskasepticallyfilledproduct,initialpositivesterilitytestresultsandinvestigationsshould

30、bereviewed.Itisdifficultforthemanufacturertojustifythereleaseofaproductfilledasepticallythatfailsaninitialsterilitytestwithoutidentifyingspecificproblemsassociatedwiththecontrolsusedforthesterilitytest.Examinetheuseofnegativecontrols.Theyareparticularlyimportanttoahighqualitysterilitytest.Goodpracti

31、ceforsuchtestingincludestheuseofknownterminallysterilizedorirradiatedsamplesasasystemcontrol.Alternatively,vialsorampulesfilledduringmediafillshavealsobeenused.Beespeciallyconcernedaboutthecasewhereamanufacturerofasepticallyfilledproductshasneverfoundaninitialpositivesterilitytest.Whilesuchsituation

32、smayoccur,theyarerare.Inonecase,amanufacturersrecordsshowedthattheyhadneverfoundapositiveresult;theirrecordshadbeenfalsified.Also,theabsenceofinitialpositivesmayindicatethatthetesthasnotbeenvalidatedtodemonstratethatthereisnocarryoverofinhibitionfromtheproductorpreservative.Inspectroboticsystemsoris

33、olationtechnology,suchasLaCalheneunitsusedforsterilitytesting.Theseunitsallowproductwithdrawalintheabsenceofpeople.Ifaninitialtestfailureisnotedinasampletestedinsuchasystem,itcouldbeverydifficulttojustifyreleasebasedonaretest,particularlyiftestcontrolsarenegative.Evaluatethetimeperiodusedforsterilit

34、ytestsampleincubation.Thisissuehasbeenrecentlyclarified.TheUSPstatesthatsamplesaretobeincubatedforatleast7days,andaproposalhasbeenmadetochangetheUSPtorequireaperiodof14daysincubation.Youareexpectedtoevaluatethespecificanalyticalprocedureandtheproductfortheproperincubationperiod.Sevendaysmaybeinsuffi

35、cient,particularlywhenslowgrowingorganismshavebeenidentified.Mediafill,environmental,sterilitytestresultsandotherdatashouldbereviewedtoassuretheabsenceofslowgrowingorganisms.Also,youshouldcomparethemethodsbeingusedforincubationtodetermineiftheyconformtothoselistedinapprovedorpendingapplications.V.ME

36、THODOLOGYANDVALIDATIONOFTESTPROCEDURESDeterminethesourceoftestprocedures.Manufacturersderivetestproceduresfromseveralsources,includingtheUSP,BAMandothermicrobiologicalreferences.Itwouldbevirtuallyimpossibletocompletelyvalidatetestproceduresforeveryorganismthatmaybeobjectionable.However,itisagoodprac

37、ticetoassurethatinhibitorysubstancesinsamplesareneutralized.Duringinspections,includingpre-approvalinspections,evaluatethemethodologyformicrobiologicaltesting.Forexample,weexpecttestmethodstoidentifythepresenceoforganismssuchasPseudomonascepaciaorotherPseudomonasspeciesthatmaybeobjectionalorpresenta

38、hazardtotheuser.Wherepre-approvalinspectionsarebeingconducted,comparethemethodbeingusedagainsttheonesubmittedintheapplication.Alsoverifythatthelaboratoryhastheequipmentnecessarytoperformthetestsandthattheequipmentwasavailableandingoodoperatingconditiononthedatesofcriticaltesting.TheUSPstatesthatanal

39、ternatemethodmaybesubstitutedforcompendialtests,providedithasbeenproperlyvalidatedasgivingequivalentorbetterresults.Youmayfindthatdehydratedmediaarebeingusedforthepreparationofmedia.Goodpracticeincludestheperiodicchallengeofpreparedmediawithlowlevelsoforganisms.ThisincludesUSPindicatororganismsaswel

40、lasnormalflora.Thecapabilityofthemediatopromotethegrowthoforganismsmaybeaffectedbythemediapreparationprocess,sterilization(overheating)andstorage.Theserepresentimportantconsiderationsinanyinspectionandinthegoodmanagementofamicrobiologylaboratory.VI.DATASTORAGEEvaluatethetestresultsthathavebeenentere

41、dineitherlogbooksoronlooseanalyticalsheets.Whilesomemanufacturersmaybereluctanttoprovidetabulations,summaries,orprintoutsofmicrobiologicaltestresults,thisdatashouldbereviewedfortheidentificationofpotentialmicrobialproblemsinprocessing.Whensummariesofthisdataarenotavailabletheinspectionteamisexpected

42、toreviewenoughdatatoconstructtheirownsummaryofthelaboratorytestresultsandqualitycontrolprogram.Somelaboratoriesutilizepreprintedformsonlyforrecordingtestdata.Somelaboratorieshavealsopointedoutthattheonlywaymicrobiologicaltestdatacouldbereviewedduringinspectionswouldbetoreviewindividualbatchrecords.H

43、owever,inmostcases,preprintedformsareinmultiplecopieswithasecondorthirdcopyinacentralfile.Somecompaniesuselog-booksforrecordingdata.Theselogbooksshouldalsobereviewed.Additionally,manymanufacturersareequippedwithanautomatedmicrobialsystemfortheidentificationofmicroorganisms.Logsofsuchtesting,alongwit

44、htheidentificationofthesourceofthesample,arealsoofvalueintheidentificationofpotentialmicrobialproblemsinprocessing.Theutilizationofautomatedsystemsfortheidentificationofmicroorganismsisrelativelycommonintheparenteralmanufacturerwhereisolatesfromtheenvironment,watersystems,validationandpeoplearerouti

45、nelyidentified.MicrobiologistsinourBaltimoreDistrictareexpertontheuseofautomatedmicrobicanalyticalsystems.TheywerethefirstFDAlaboratorytousesuchequipmentandhaveconsiderableexperienceinvalidatingmethodsforthesepiecesofequipment.ContacttheBaltimoreDistrictlaboratoryforinformationorquestionsabouttheses

46、ystems.PlantswithheavyutilizationofthesepiecesofequipmentshouldbeinspectedbyindividualsfromtheBaltimoreDistrictlaboratory.VII.MANAGEMENTREVIEWMicrobiologicaltestresultsrepresentoneofthemoredifficultareasfortheevaluationandinterpretationofdata.Theseevaluationsrequireextensivetrainingandexperienceinmi

47、crobiology.Understandingthemethodology,andmoreimportantly,understandingthelimitationsofthetestpresentthemoredifficultissues.Forexample,amanufacturerfoundhighcountsofEnterobactercloacaeintheiroraldosageformproductderivedfromanaturalsubstance.SincetheydidnotisolateE.coli,theyreleasedtheproduct.FDAanalysisfoundE.cloacaeinmostsamplesfromthebatchandevenE.coliinonesample.Inthiscasemanagementfailedtorecognizethatmicrobiologicalcontaminationmightnotbeuniform,thatotherorganismsmaymaskthepresenceofcertainorganismswhenidentificationproceduresareperformed,andthatmicrobiologicaltestingisfarfroma