USP71 无菌检查法Sterility Tests(1)

USP<71>无菌检查法SterilityTests无菌检查法系用于检查药典要求无菌的药物、制剂产品和其他物品是否无菌的一种 方法 快递客服问题件处理详细方法山木方法pdf计算方法pdf华与华方法下载八字理论方法下载 。若供试品符合无菌检查法的规定，仅 表 关于同志近三年现实表现材料材料类招标技术评分表图表与交易pdf视力表打印pdf用图表说话 pdf 明供试品在该检验条件下未发现微生物污染。Thetestisappliedtosubstances,preparations,orarticleswhich,accordingtothePharmacopeia,arerequiredtobesterile.However,asatisfactoryresultonlyindicatesthatnocontaminatingmicroorganismhasbeenfoundinthesampleexaminedundertheconditionsofthetest.1预防微生物污染PrecaotionsAgainstMicrobialContamination无菌检查应在无菌条件下进行，为了达到条件，检测环境应符合无菌检查的规定。防止污染的措施不得影响供试品中微生物的检出。检测环境应定期抽样监测并进行适当控制。Thetestforsterilityiscarriedoutunderasepticconditions.Inordertoachievesuchconditions,thetestenvironmenthastobeadaptedtothewayinwhichthesterilitytestisperformed.Theprecautionstakentoavoidcontaminationaresuchthattheydonotaffectanymicroorganismsthataretoberevealedinthetest.Theworkingconditionsinwhichthetestsareperformedaremonitoredregularlybyappropriatesamplingoftheworkingareaandbycarryingoutappropriatecontrols.2培养基和培养温度CultureMediaandIncubation无菌检查需制备下表所述培养基，或者是能够符合需氧菌、厌氧菌、真菌促生长试验要求的同等的商用培养基。MediaforthetestmaybepreparedasdescribedbeloworequivalentcommercialmediamaybeusedprovidedthattheycomplywiththerequirementsoftheGrowthPromotionTestofAerobes,Anaerobes,andFungi.以下培养基已经被证实适合用于无菌检查。硫乙醇酸盐流体培养基主要用于厌氧菌的培养，但其也可用于需氧菌培养。大豆酪蛋白培养基适合于培养真菌和需氧菌。Thefollowingculturemediahavebeenfoundtobesuitableforthetestforsterility.FluidThioglycollateMediumisprimarilyintendedforthecultureofanaerobicbacteria.However,itwillalsodetectaerobicbacteria.Soybean–CaseinDigestMediumissuitableforthecultureofbothfungiandaerobicbacteria.将L-胱氨酸、氯化钠、葡萄糖、酵母提取物、酪蛋白胰酶消化物与纯化水混合，并加热至溶解。将硫乙醇酸钠或硫乙醇酸溶解于该溶液，如果需要可再加入1mol/L氢氧化钠溶液，以便在灭菌后该溶液呈pH值7.1±0.2。如需要过滤，再次加热该溶液但不得煮沸，并趁热以润湿滤纸将该溶液过滤。加入刃天青钠溶液，混匀，并将该培养基置于适当容器中，该容器应为培养基提供特定的面积/深度比，以使在培养期结束后能明确显示氧气摄入的变色部分不超过培养基的上半部分二分之一。使用经过验证的工艺进行灭菌。如果需要贮存该培养基，应将其置于无菌、气密容器中，在2~25℃贮藏。如果超过上部三分之一的培养基已经呈粉红色，可以用以下方法恢复该培养基功能，但每批培养基仅能恢复一次：在水浴锅中或者自由流动蒸汽中加热该容器，直至粉色消失，并迅速放凉，须小心防止非无菌空气进入到容器中。灭菌后培养基存放时间超过验证期限时，不得使用。MixtheL-cystine,agar,sodiumchloride,dextrose,yeastextract,andpancreaticdigestofcaseinwiththepurifiedwater,andheatuntilsolutioniseffected.Dissolvethesodiumthioglycollateorthioglycolicacidinthesolutionand,ifnecessary,add1Nsodiumhydroxidesothat,aftersterilization,thesolutionwillhaveapHof7.1±0.2.Iffiltrationisnecessary,heatthesolutionagainwithoutboiling,andfilterwhilehotthroughmoistenedfilterpaper.Addtheresazurinsodiumsolution,mix,andplacethemediuminsuitablevesselsthatprovidearatioofsurfacetodepthofmediumsuchthatnotmorethantheupperhalfofthemediumhasundergoneacolorchangeindicativeofoxygenuptakeattheendoftheincubationperiod.Sterilizeusingavalidatedprocess.Ifthemediumisstored,storeatatemperaturebetween2℃and25℃inasterile,airtightcontainer.Ifmorethantheupperone-thirdofthemediumhasacquiredapinkcolor,themediummayberestoredoncebyheatingthecontainersinawater-bathorinfree-flowingsteamuntilthepinkcolordisappearsandbycoolingquickly,takingcaretopreventtheintroductionofnonsterileairintothecontainer.Donotusethemediumforalongerstorageperiodthanhasbeenvalidated.硫乙醇酸盐流体培养基FluidThioglycollateMediumL-胱氨酸L-cystine0.5g氯化钠SodiumChloride2.5g水合葡萄糖/无水葡萄糖DextroseMonohydrate/Anhydrous5.5/5.0g琼脂Agar0.75g酵母提取物（水溶的）YeastExtract(water-soluble)5.0g胰酶消化酪蛋白胨PancreaticDigestofCasein15.0g硫乙醇酸钠SodiumThioglycollate0.5g或硫乙醇酸orThioglycolicAcid0.3ml新配制的刃天青水溶液（1:1000）ResazurinSodiumSolution(1in1000),freshlyprepared1.0ml纯化水PurifiedWater1000ml灭菌后pH：7.1±0.2pHaftersterilization:7.1±0.2.硫乙醇酸盐流体培养基应在30~35℃条件下进行培养。含有汞制剂防腐剂的产品不能使用膜过滤方法检测。经需氧菌、厌氧菌、真菌促生长试验验证，在20~25℃培养时，大豆酪蛋白消化培养基可以替代硫乙醇酸盐流体培养基。经合理授权，不含琼脂和刃天青钠溶液配制的硫乙醇酸盐流体培养基可以替代硫乙醇酸盐流体培养基使用。按上述方法灭菌，灭菌后pH值为7.1±0.2。使用前使用水浴加热，置于30~35℃厌氧条件下培养。FluidThioglycollateMediumistobeincubatedat30–35℃.Forproductscontainingamercurialpreservativethatcannotbetestedbythemembranefiltrationmethod,FluidThioglycollateMediumincubatedat20–25℃maybeusedinsteadofSoybean–CaseinDigestMediumprovidedthatithasbeenvalidatedasdescribedinGrowthPromotionTestofAerobes,Anaerobes,andFungi.Whereprescribedorjustifiedandauthorized,thefollowingalternativethioglycollatemediummightbeused.PrepareamixturehavingthesamecompositionasthatoftheFluidThioglycollateMedium,butomittingtheagarandtheresazurinsodiumsolution.Sterilizeasdirectedabove.ThepHaftersterilizationis7.1±0.2.Heatinawaterbathpriortouseandincubateat30–35℃underanaerobicconditions.大豆酪蛋白消化物培养基Soybean–CaseinDigestMedium酪蛋白胰酶消化物PancreaticDigestofCasein17.0g大豆粉木瓜蛋白酶消化物PapaicDigestofSoybeanMeal3.0g氯化钠SodiumChloride5.0g磷酸氢二钾DibasicPotassiumPhosphate2.5g水合葡萄糖/无水DextroseMonohydrate/Anhydrous2.5/2.3g纯化水PurifiedWater1000ml灭菌后pH：7.3±0.2pHaftersterilization:7.3±0.2.将干粉培养基置纯化水中，轻微加热使其溶解。溶液放凉至室温，并用1mol/L氢氧化钠溶液调整pH值，使其灭菌后pH值为7.3±0.2。如需要使之澄清，则过滤，分装入适合的容器，并用经过验证的程序灭菌。如果不立刻使用，则保存在2~25℃无菌且密闭良好的容器中。灭菌后培养基存放时间超过验证期限时，不得使用。DissolvethesolidsinthePurifiedWater,heatingslightlytoeffectasolution.Coolthesolutiontoroomtemperature,andadjustthepHwith1Nsodiumhydroxidesothat,aftersterilization,itwillhaveapHof7.3±0.2.Filter,ifnecessarytoclarify,dispenseintosuitablecontainers,andsterilizeusingavalidatedprocedure.Storeatatemperaturebetween2℃and25℃inasterilewell-closedcontainer,unlessitisintendedforimmediateuse.Donotusethemediumforalongerstorageperiodthanhasbeenvalidated.大豆酪蛋白消化物培养基应在22.5℃±2.5℃条件下培养。Soybean–CaseinDigestMediumistobeincubatedat22.5℃±2.5℃.2.1用于青霉素和头孢菌素的培养基MediaforPenicillinsorCephalosporins当无菌检查培养基用于供试产品无菌检查项下的直接接种法检验时，按如下内容变更硫乙醇酸盐流体培养基和大豆酪蛋白培养基的制备方法。向每一种培养基的容器中，以无菌操作转移足够量能灭活供试样品中抗生素的β-内酰胺酶（之前已经对β-内酰胺酶制品灭活青霉素或头孢菌素的能力进行过测定，据此来确定灭活该抗生素所需的β-内酰胺酶量）（注：添加β-内酰胺酶的培养基也可以用于膜过滤试验）WheresterilitytestmediaaretobeusedintheDirectInoculationoftheCultureMediummethodunderTestforSterilityoftheProducttobeExamined,modifythepreparationofFluidThioglycollateMediumandtheSoybean–CaseinDigestMediumasfollows.Tothecontainersofeachmedium,transferasepticallyaquantityofβ-lactamasesufficienttoinactivatetheamountofantibioticinthespecimenundertest.Determinethequantityofβ-lactamaserequiredtoinactivatetheantibioticbyusingaβ-lactamasepreparationthathasbeenassayedpreviouslyforitspenicillin-orcephalosporin-inactivatingpower.[Note—Supplementedβ-lactamasemediacanalsobeusedinthemembranefiltrationtest.]表1用于促生长试验和方法验证试验的测试菌株Table1.StrainsoftheTestMicroorganismsSuitableforUseintheGrowthPromotionTestandtheMethodSuitabilityTest需氧菌Aerobicbacteria金黄色葡萄球菌StaphylococcusaureusATCC6538,CIP4.83,NCTC10788,NCIMB9518,NBRC13276枯草芽孢杆菌BacillussubtilisATCC6633,CIP52.62,NCIMB8054,NBRC3134铜绿假单胞菌Pseudomonasaeruginosa①ATCC9027,NCIMB8626,CIP82.118,NBRC13275厌氧菌Anaerobicbacterium生孢梭菌Clostridiumporogenes②ATCC19404,CIP79.3,NCTC532orATCC11437,NBRC14293真菌Fungi白色念珠菌CandidaalbicansATCC10231,IP48.72,NCPF3179,NBRC14293巴西曲霉Aspergillusbrasiliensis(Aspergillusniger)（黑曲霉）ATCC16404,IP1431.83,IMI149007,NBRC9455①替代微生物是藤黄微球菌（KocuriarhizopHila,ATCC9341）AnalternativemicroorganismisKocuriarhizophila(Micrococcusluteus)ATCC9341.②需要不形成芽孢微生物时，生孢梭菌的替代替代微生物是普通拟杆菌（Bacetroidesvulgatus,ATCC8482）AnalternativetoClostridiumsporogenes,whenanonspore-formingmicroorganismisdesired,isBacteroidesvulgatus(ATCC8482).所使用的培养基须符合下列试验，这些试验应在供试产品检验前完成或者同时进行。Themediausedcomplywiththefollowingtests,carriedoutbefore,orinparallel,withthetestontheproducttobeexamined.3培养基无菌性SterilityofCultureMedia取部分检验用培养基，连续培养14天，应不得出现微生物生长。Incubateportionsofthemediafor14days.Nogrowthofmicroorganismsoccurs.3.1需气菌、厌氧菌和真菌的促生长试验GrowthPromotionTestofAerobes,Anaerobes,andFungi每一批配制好的培养基（采用脱水培养基或按配方制备的培养基）均需进行检查，使用的微生物菌株见表1。Testeachlotofready-preparedmediumandeachbatchofmediumpreparedeitherfromdehydratedmediumorfromingredients.SuitablestrainsofmicroorganismsareindicatedinTable1.取部分硫乙醇酸盐流体培养基，接种少量（不超过100cfu）下列微生物，每一种微生物均分别接种于单独培养基中：生孢梭菌、铜绿假单胞菌、金黄色葡萄球菌（在替代硫乙醇酸盐的流体培养基中接种少量（不超过100cfu）生孢梭菌）。在大豆酪蛋白消化物培养基中接种少量（不超过100cfu）下列微生物，每一种微生物均接种于单独的培养基：黑曲霉、枯草芽孢杆菌、白色念珠菌。细菌培养时间不超过3天，真菌培养时间不超过5天。采用适宜的菌种保藏技术，以确保用于接种的微生物的传达次数不超过5代。InoculateportionsofFluidThioglycollateMediumwithasmallnumber(notmorethan100cfu)ofthefollowingmicroorganisms,usingaseparateportionofmediumforeachofthefollowingspeciesofmicroorganism:Clostridiumsporogenes,Pseudomonasaeruginosa,andStaphylococcusaureus.Inoculateportionsofalternativethioglycollatemediumwithasmallnumber(notmorethan100cfu)ofClostridiumsporogenes.InoculateportionsofSoybean–CaseinDigestMediumwithasmallnumber(notmorethan100cfu)ofthefollowingmicroorganisms,usingaseparateportionofmediumforeachofthefollowingspeciesofmicroorganism:Aspergillusbrasiliensis,Bacillussubtilis,andCandidaalbicans.Incubatefornotmorethan3daysinthecaseofbacteriaandnotmorethan5daysinthecaseoffungi.Seedlotculturemaintenancetechniques(seed-lotsystems)areusedsothattheviablemicroorganismsusedforinoculationarenotmorethanfivepassagesremovedfromtheoriginalmasterseed-lot.如果可见清晰的微生物生长，则该培养基是符合要求的。Themediaaresuitableifaclearlyvisiblegrowthofthemicroorganismsoccurs.3.2用于膜过滤的稀释剂和冲洗液DilutionandRinsingFluidsforMembraneFiltration3.2.1稀释剂AFluidA3.2.1.1制备Preparation将1g动物组织胃蛋白酶消化物溶于1L水中，如果需要则通过过滤或离心使其澄清，再调节pH值至7.1±0.2。分装入容器中，并经过验证的工艺灭菌。Dissolve1gofpepticdigestofanimaltissueinwatertomake1L,filterorcentrifugetoclarify,ifnecessary,andadjusttoapHof7.1±0.2.Dispenseintocontainers,andsterilizeusingavalidatedprocess.3.2.1.2用于青霉素或头孢菌素的稀释剂制备PreparationforPenicillinsorCephalosporins在供试样品溶液已经过滤之后，如果需要，以无菌操作向上述制备的稀释剂中加入数量足够灭活滤膜上残余抗生素的β-内酰胺（见用于青霉素或头孢菌素的培养基）。AsepticallyaddtotheabovePreparation,ifnecessary,aquantityofsterileβ-lactamasesufficienttoinactivateanyresidualantibioticactivityonthemembranesafterthesolutionofthetestspecimenhasbeenfiltered(seeMediaforPenicillinsorCephalosporins).3.2.2稀释剂DFluidD向每升稀释剂A中，加入1ml聚山梨酯80，调节pH值至7.1±0.2，分装入容器中，并使用经过验证的工艺灭菌。此液体用于含有卵磷脂或油脂的样品，或用于标为“无菌通道（SterilePathway）”的器械。ToeachLofFluidAadd1mLofpolysorbate80,adjusttoapHof7.1±0.2,dispenseintocontainers,andsterilizeusingavalidatedprocess.Usethisfluidforarticlescontaininglecithinoroil,orfordeviceslabeledas“sterilepathway.”3.2.3稀释剂KFluidK将5.0g动物组织胃蛋白酶消化物、3.0g牛肉提取物、10.0g聚山梨酯80溶解于1L水中。调节pH值为6.9±0.2。分装入容器中，并使用经过验证的工艺灭菌。Dissolve5.0gofpepticdigestofanimaltissue,3.0gofbeefextract,and10.0gofpolysorbate80inwatertomake1L.AdjustthepHtoobtain,aftersterilization,apHof6.9±0.2.Dispenseintocontainers,andsterilizeusingavalidatedprocess.4方法适用性实验MethodSuitabilityTest严格按照供试产品无菌检查项下的方法进行无菌检查。当使用到以下方法并需要进行方法调整时，需重新进行方法适用性实验。CarryoutatestasdescribedbelowunderTestforSterilityoftheProducttobeExaminedusingexactlythesamemethods,exceptforthefollowingmodifications.4.1膜过滤法MembraneFiltration在将一个或多个供试容器中的内容物转移到滤膜之后，在最后一次的冲洗液中加入少量（不超过100cfu）测试菌株。Aftertransferringthecontentofthecontainerorcontainerstobetestedtothemembrane,addaninoculumofasmallnumberofviablemicroorganisms(notmorethan100cfu)tothefinalportionofsterilediluentusedtorinsethefilter.4.2直接接种法DirectInoculation在将一个或多个供试容器（兽医用的肠线和其他外科缝合用线：若干股线）中的内容物转移至培养基之后，将少量试验菌（不超过100cfu）加入至培养基中。Aftertransferringthecontentsofthecontainerorcontainerstobetested(forcatgutandothersurgicalsuturesforveterinaryuse:strands)totheculturemedium,addaninoculumofasmallnumberofviablemicroorganisms(notmorethan100cfu)tothemedium.以上两种情况，均使用上述需氧菌、厌氧菌、真菌促生长试验项下规定的菌株。同时设置促生长试验作为阳性对照。微生物在相应培养基中的培养时间不超过5天。InbothcasesusethesamemicroorganismsasthosedescribedaboveunderGrowthPromotionTestofAerobes,Anaerobes,andFungi.Performagrowthpromotiontestasapositivecontrol.Incubateallthecontainerscontainingmediumfornotmorethan5days.若培养后可见清晰的微生物生长，外观与未接种样品的对照菌生长类似，则认为该产品在此试验条件下没有抑菌作用，或者认为可能存在的抑菌作用已经被较完全地消除。此时可以认为该方法无需进一步的变更，无菌试验依法检查即可。Ifclearlyvisiblegrowthofmicroorganismsisobtainedaftertheincubation,visuallycomparabletothatinthecontrolvesselwithoutproduct,eithertheproductpossessesnoantimicrobialactivityundertheconditionsofthetestorsuchactivityhasbeensatisfactorilyeliminated.Thetestforsterilitymaythenbecarriedoutwithoutfurthermodification.与未接种样品的对照相比，若在供试品存在条件下不能观察到肉眼可见的混浊，则认为该试验条件不能消除供试品的抑菌作用，需要对实验条件进行调整以消除抗菌活性，并重新进行方法适用性试验。Ifclearlyvisiblegrowthisnotobtainedinthepresenceoftheproducttobetested,visuallycomparabletothatinthecontrolvesselswithoutproduct,theproductpossessesantimicrobialactivitythathasnotbeensatisfactorilyeliminatedundertheconditionsofthetest.Modifytheconditionsinordertoeliminatetheantimicrobialactivity,andrepeattheMethodSuitabilityTest.当一个新产品进行无菌试验时和无菌试验的试验条件发生改变时，则需进行验证试验，该验证可以与供试产品无菌检查同时进行。Thismethodsuitabilityisperformedwhenthetestforsterilityhastobecarriedoutonanewproduct;andwheneverthereisachangeintheexperimentalconditionsofthetest.ThemethodsuitabilitymaybeperformedsimultaneouslywiththeTestforSterilityoftheProducttobeexamined.5供试产品无菌检查TestforSterilityoftheProducttobeExamined5.1供试品数量NumberofArticlestoBeTested除另有规定外，供试品的数量遵照表3中的规定。如果每个被检验物品的内容物有足够数量（见表2），可以将其分成若干等分，将适当的等份加入到每个指定的培养基。（注：使用两个或更多指定培养基，来进行无菌试验。）如果每个被检验物品的内容物 规格 视频线规格配置磁共振要求常用水泵型号参数扭矩规格钢结构技术规格书 不能满足单个培养基的接种量要求时，检验量扩大为表3所规定的检验数量的2倍。Unlessotherwisespecifiedelsewhereinthischapterorintheindividualmonograph,testthenumberofarticlesspecifiedinTable3.Ifthecontentsofeacharticleareofsufficientquantity(seeTable2),theymaybedividedsothatequalappropriateportionsareaddedtoeachofthespecifiedmedia.[Note—Performsterilitytestingemployingtwoormoreofthespecifiedmedia.]Ifeacharticledoesnotcontainsufficientquantitiesforeachmedium,usetwicethenumberofarticlesindicatedinTable3.表2每种培养基的最少接种量Table2.MinimumQuantitytobeUsedforEachMedium供试品装量QuantityperContainer最少接种量（除另有规定和授权）MinimumQuantitytobeUsed(unlessotherwisejustifiedandauthorized)液体Liquids少于1mlLessthan1mL每个包装全量Thewholecontentsofeachcontainer1~40ml每个包装半量，且不少于1mlHalfthecontentsofeachcontainer,butnotlessthan1mL大于40ml，但不大于100mlGreaterthan40mL,andnotgreaterthan100mL20ml大于100mlGreaterthan100mL每个包装内容物的10%，但不得少于20ml10%ofthecontentsofthecontainer,butnotlessthan20mL液体抗生素Antibioticliquids1ml需悬浮或乳化的不溶性制剂，乳膏、油膏Insolublepreparations,creams,andointmentstobesuspendedoremulsified每个包装不少于200mg内容物Usethecontentsofeachcontainertoprovidenotlessthan200mg固体Solids少于50mgLessthan50mg每个包装全量Thewholecontentsofeachcontainer50mg或以上，但不大于300mg50mgormore,butlessthan300mg每个包装半量，但不少于50mgHalfthecontentsofeachcontainer,butnotlessthan50mg300mg~5g150mg多于5gGreaterthan5g500mg兽医用肠线和其他外科缝合线Catgutandothersurgicalsuturesforveterinaryuse一股线的3部分（每个30cm长）3sectionsofastrand(each30cmlong)外科敷料/棉花/纱布（在包装中）Surgicaldressing/cotton/gauze(inpackages)每包装100mg100mgperpackage缝合线和其他独立包装的一次性材料Suturesandotherindividuallypackagedsingle-usematerial材料整体Thewholedevice其他医用设备Othermedicaldevices整个设备，切成片或拆开Thewholedevice,cutintopiecesordisassembled表3批出厂产品的最小检验数量Table3.MinimumNumberofArticlestobeTestedinRelationtotheNumberofArticlesintheBatch批产品数量NumberofItemsintheBatch最小样品数量（除另有规定和授权）MinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)注射剂Parenteralpreparations不多于100个容器Notmorethan100containers10%或4个容器，选较多者10%or4containers,whicheveristhegreater多于100个，但不多于500个容器Morethan100butnotmorethan500containers10个容器10containers多于500个容器Morethan500containers2%或者20个容器，选较少者2%or20containers,whicheverisless大体积注射剂Forlarge-volumeparenterals2%或者10个容器，选较少者2%or10containers,whicheverisless固体抗生素Antibioticsolids固体制剂（＜5g）Pharmacybulkpackages(<5g)20个容器20containers固体制剂（≥5g）Pharmacybulkpackages(≥5g)6个容器6containers散装固体Bulksandblends见固体原料SeeBulksolidproducts眼科和其他非注射剂Ophthalmicandothernoninjectablepreparations不多于200个容器Notmorethan200containers5%或2个容器，选较多者5%or2containers,whicheveristhegreater多于200个容器Morethan200containers10个容器10containers如果该产品存在于单剂量容器中，应用上述用于注射剂 方案 气瓶 现场处置方案 .pdf气瓶 现场处置方案 .doc见习基地管理方案.doc关于群访事件的化解方案建筑工地扬尘治理专项方案下载 Iftheproductispresentedintheformofsingle-dosecontainers,applytheschemeshownaboveforpreparationsforparenteraluse.兽医用肠线和其他外科缝合线Catgutandothersurgicalsuturesforveterinaryuse2%或5个包装，选较多者，最多可达20个包装2%or5packages,whicheveristhegreater,uptoamaximumtotalof20packages不多于100件Notmorethan100articles10%或4个物品，选较多者10%or4articles,whicheverisgreater多于100但不超过500件Morethan100,butnotmorethan500articles10个物品10articles多于500件Morethan500articles2%或20个物品，选较少者2%or20articles,whicheverisless固体原料Bulksolidproducts最多4个容器Upto4containers每个容器Eachcontainer多于4个容器，但不多于50个容器Morethan4containers,butnotmorethan50containers20%或4个容器，选较多者20%or4containers,whicheverisgreater超过50个容器Morethan50containers2%或10个容器，选较多者2%or10containers,whicheverisgreater如果批量大小未知，请使用相关规定的最大数Ifthebatchsizeisunknown,usethemaximumnumberofitemsprescribed.如果一个容器的内容物足够接种2个培养基，则此表格给出的容器数量为用于全部2个培养基的数量Ifthecontentsofonecontainerareenoughtoinoculatethetwomedia,thiscolumngivesthenumberofcontainersneededforboththemediatogether.无菌检验可以使用膜过滤法或培养基直接接种法，应设置适当的阴性对照。但只要该产品的性质许可，就应使用膜过滤法；也就是说，对水溶性制剂、酒精或油性制剂、易混合或溶解于水或油性溶剂的制剂，即使没有抑菌作用，也均应尽量采用膜过滤法进行无菌检查。ThetestmaybecarriedoutusingthetechniqueofMembraneFiltrationorbyDirectInoculationoftheCultureMediumwiththeproducttobeexamined.Appropriatenegativecontrolsareincluded.Thetechniqueofmembranefiltrationisusedwheneverthenatureoftheproductpermits;thatis,forfilterableaqueouspreparations,foralcoholicoroilypreparations,andforpreparationsmisciblewith,orsolublein,aqueousoroilysolvents,providedthesesolventsdonothaveanantimicrobialeffectintheconditionsofthetest.5.2膜过滤法MembraneFiltration使用标称孔径不大于0.45μm的膜过滤器，此孔径已知能够有效截留微生物。例如，硝酸纤维素过滤器可用于水、油、稀醇溶液；而醋酸纤维素可用于浓醇溶液。特定产品（例如抗生素）可能需要特别改造过的特殊过滤器。Usemembranefiltershavinganominalporesizenotgreaterthan0.45μm,inwhichtheeffectivenesstoretainmicroorganismshasbeenestablished.Cellulosenitratefilters,forexample,areusedforaqueous,oily,andweaklyalcoholicsolutions;andcelluloseacetatefilters,forexample,areusedforstronglyalcoholicsolutions.Speciallyadaptedfiltersmaybeneededforcertainproducts(e.g.,forantibiotics).以下涉及的方法中所使用的均为直径约50mm的滤膜。如果使用不同直径的过滤器，稀释液和冲洗液的体积应当作相应调节。过滤器和滤膜都应该经过灭菌处理。过滤器应能在无菌状态下摘掉滤膜并将其转移至培养基中，或应能将培养基灌注至该设备中进行培养。Thetechniquedescribedbelowassumesthatmembranesabout50mmindiameterwillbeused.Iffiltersofadifferentdiameterareused,thevolumesofthedilutionsandthewashingsshouldbeadjustedaccordingly.Thefiltrationapparatusandmembranearesterilizedbyappropriatemeans.Theapparatusisdesignedsothatthesolutiontobeexaminedcanbeintroducedandfilteredunderasepticconditions:itpermitstheasepticremovalofthemembranefortransfertothemedium,oritissuitableforcarryingouttheincubationafteraddingthemediumtotheapparatusitself.5.2.1水溶性溶液AqueousSolutions应当尽量减少转移至过滤器滤膜上的无菌稀释剂，例如稀释剂A（见用于膜过滤的稀释剂和冲洗液）。稀释剂中可能会含有一定量的中和物质或灭活物质，例如对抗生素的检测。Ifappropriate,transferasmallquantityofasuitable,sterilediluentsuchasFluidA(seeDilutingandRinsingFluidsforMembraneFiltration)ontothemembraneintheapparatusandfilter.Thediluentmaycontainsuitableneutralizingsubstancesand/orappropriateinactivatingsubstances,forexample,inthecaseofantibiotics.将一个或多个供试容器的内容物转移到滤膜，并立即过滤。如需要可先用选定的无菌稀释剂稀释至方法适用性试验中所用的体积，但须使用不少于表2和表3中规定的供试品数量。如果该产品具有抗菌活性，滤膜至少冲洗3次，每次冲洗量均按照方法适用性试验中确定的无菌稀释剂体积冲洗滤膜。即便该冲洗循环不能完全消除抗菌活性，冲洗过程通常不应该超过“5次，100ml/次”。将整个滤膜转移至培养基，或以无菌操作将滤膜切至相等的2部分，并将每一部分转移至适当的培养基中。每个培养基的体积按照方法适用性试验所确定的用量，或者，将培养基转移至带有滤膜的滤器中。培养该培养基，不少于14天。Transferthecontentsofthecontainerorcontainerstobetestedtothemembraneormembranes,ifnecessary,afterdilutingtothevolumeusedintheMethodSuitabilityTestwiththechosensterilediluent,butusingnotlessthanthequantitiesoftheproducttobeexaminedprescribedinTables2and3.Filterimmediately.Iftheproducthasantimicrobialproperties,washthemembranenotlessthanthreetimesbyfilteringthroughiteachtimethevolumeofthechosensterilediluentusedintheMethodSuitabilityTest.Donotexceedawashingcycleoffivetimes100mLperfilter,evenifduringmethodsuitabilityithasbeendemonstratedthatsuchacycledoesnotfullyeliminatetheantimicrobialactivity.Transferthewholemembranetotheculturemediumorcutitasepticallyintotwoequalparts,andtransferonehalftoeachoftwosuitablemedia.UsethesamevolumeofeachmediumasintheMethodSuitabilityTest.Alternatively,transferthemediumontothemembraneintheapparatus.Incubatethemediafornotlessthan14days.5.2.2可溶性固体SolubleSolids在每个培养基中，使用不少于表2和表3规定的产品数量溶于适当溶剂，例如稀释剂A（用于膜过滤的稀释剂和冲洗液）并按照上述关于水溶性溶液样品实验操作的要求，使用适合所选溶剂的滤膜，进行试验。UseforeachmediumnotlessthanthequantityprescribedinTables2and3oftheproductdissolvedinasuitablesolvent,suchasthesolventprovidedwiththepreparation,SterileWaterforInjection,sterilesaline,orasuitablesterilesolutionsuchasFluidA(DilutingandRinsingFluidsforMembraneFiltration),andproceedwiththetestasdescribedaboveforAqueousSolutionsusingamembraneappropriatetothechosensolvent.5.2.3油和油性溶液OilsandOilySolutions在每个培养基中，使用不少于表2和表3中规定的检验量。黏性较低的油和油性溶液可在不经稀释的情况下滤过干燥滤膜。黏稠油质可以用适合的无菌稀释剂进行稀释，如已证实在该实验条件下不具有抗菌活性的豆蔻酸异丙酯，油质依靠其自身的重量穿过滤膜，然后逐渐加压或抽吸过滤。使用适宜的无菌冲洗液冲洗滤膜，至少冲洗3次，约100ml/次，例如使用一定浓度乳化剂的稀释剂A（参见用于膜过滤的稀释剂和冲洗液）。通过方法适用性试验确定乳化剂的适宜浓度，例如浓度为每升10克的聚山梨