美国药典USP无菌检查

（医疗药品管理）美国药典USP无菌检查美国药典USP31-NF26无菌检查法《71》.doc71STERILITYTESTS无菌检查法PortionsofthisgeneralchapterhavebeenharmonizedwiththecorrespondingtextsoftheEuropeanPharmacopeiaand/ortheJapanesePharmacopeia.Thoseportionsthatarenotharmonizedaremarkedwithsymbols()tospecifythisfact.此通则的各部分已经与欧洲药典和/或日本药典的对应部分做了协调。不一致的部分用符号（）来标明。ThefollowingproceduresareapplicablefordeterminingwhetheraPharmacopeialarticlepurportingtobesterilecomplieswiththerequirementssetforthintheindividualmonographwithrespecttothetestforsterility.PharmacopeialarticlesaretobetestedbytheMembraneFiltrationmethodunderTestforSterilityoftheProducttobeExaminedwherethenatureoftheproductpermits.Ifthemembranefiltrationtechniqueisunsuitable,usetheDirectInoculationoftheCultureMediummethodunderTestforSterilityoftheProducttobeExamined.Alldevices,withtheexceptionofDeviceswithPathwaysLabeledSterile,aretestedusingtheDirectInoculationoftheCultureMediummethod.ProvisionsforretestingareincludedunderObservationandInterpretationofResults.下面这些步骤适用于测定是否某个用于无菌用途的药品是否符合其具体的各论中关于无菌检查的要求。只要其性质许可，这些药品将使用供试产品无菌检查法项下的膜过滤法来检测。如果膜过滤技术是不适合的，则使用在供试产品无菌检查法项下的培养基直接接种法。除了具有标记为无菌通道的设备之外，所有的设备均须使用培养基直接接种法进行检测。在结果的观测与理解项下包含了复验的规定。Becausesterilitytestingisaveryexactingprocedure,whereasepsisoftheproceduremustbeensuredforacorrectinterpretationofresults,itisimportantthatpersonnelbeproperlytrainedandqualified.Thetestforsterilityiscarriedoutunderasepticconditions.Inordertoachievesuchconditions,thetestenvironmenthastobeadaptedtothewayinwhichthesterilitytestisperformed.Theprecautionstakentoavoidcontaminationaresuchthattheydonotaffectanymicroorganismsthataretoberevealedinthetest.Theworkingconditionsinwhichthetestsareperformedaremonitoredregularlybyappropriatesamplingoftheworkingareaandbycarryingoutappropriatecontrols.由于无菌检查法是一个非常精确的程序，在此过程中程序的无菌状态必须得到确保以实现对结果的正确理解，因此人员经过适当的培训并取得资质是非常重要的。无菌检查在无菌条件下进行。为了实现这样的条件，试验环境必须调整到适合进行无菌检查的方式。为避免污染而采取的特定预防措施应不会对任何试图在检查中发现的微生物产生影响。通过在工作区域作适当取样并进行适当控制，来定期监测进行此试验的工作条件。ThesePharmacopeialproceduresarenotbythemselvesdesignedtoensurethatabatchofproductissterileorhasbeensterilized.Thisisaccomplishedprimarilybyvalidationofthesterilizationprocessoroftheasepticprocessingprocedures.这些药典规定程序自身的设计不能确保一批产品无菌或已经灭菌。这主要是通过灭菌工艺或者无菌操作程序的验证来完成。WhenevidenceofmicrobialcontaminationinthearticleisobtainedbytheappropriatePharmacopeialmethod,theresultsoobtainedisconclusiveevidenceoffailureofthearticletomeettherequirementsofthetestforsterility,evenifadifferentresultisobtainedbyanalternativeprocedure.Foradditionalinformationonsterilitytesting,seeSterilizationandSterilityAssuranceofCompendialArticles1211.当通过适当的药典方法获得了某物品中微生物污染的证据，这样获得的结果是该物品未能达到无菌检验要求的结论性证据，即便使用替代程序得到了不同的结果也无法否定此结果。如要获得关于无菌检验的其他信息，见药品的灭菌和无菌保证<1211>MEDIA培养基Preparemediaforthetestsasdescribedbelow,ordehydratedformulationsmaybeusedprovidedthat,whenreconstitutedasdirectedbythemanufacturerordistributor,theymeettherequirementsoftheGrowthPromotionTestofAerobes,Anaerobes,andFungi.Mediaaresterilizedusingavalidatedprocess.按照下面描述的方法配制实验用培养基；或者使用脱水培养基，只要根据其制造商或者分销商说明进行恢复之后，其能够符合好氧菌、厌氧菌、霉菌生长促进试验的要求即可。使用经过验证的工艺对培养基进行灭菌操作。Thefollowingculturemediahavebeenfoundtobesuitableforthetestforsterility.FluidThioglycollateMediumisprimarilyintendedforthecultureofanaerobicbacteria.However,itwillalsodetectaerobicbacteria.Soybean–CaseinDigestMediumissuitableforthecultureofbothfungiandaerobicbacteria.下面的培养基已经被证实适合进行无菌检查。巯基醋酸盐液体培养基主要用于厌氧菌的培养。但其也用于检测好氧菌。大豆酪蛋白消化物培养基适合于培养霉菌和好氧菌。FluidThioglycollateMedium巯基醋酸盐液体培养基L-CystineL-胱氨酸0.5gSodiumChloride氯化钠2.5g5.5/5.0Dextrose(C6H12O6·H2O)葡萄糖gAgar,granulated(moisturecontentnotexceeding15%)琼脂，呈颗粒状(水分含量不超过15%)0.75gYeastExtract(water-soluble)酵母提取物(水溶性)5.0gPancreaticDigestofCasein酪蛋白胰酶消化物15.0gSodiumThioglycollate巯基乙酸钠0.5gorThioglycolicAcid或者巯基乙酸0.3mLResazurinSodiumSolution(1in1000),freshlyprepared刃天青钠溶液(1比1000)，新配制1.0mLPurifiedWater纯净水1000mLMixtheL-cystine,sodiumchloride,dextrose,yeastextract,andpancreaticdigestofcaseinwiththepurifiedwater,andheatuntilsolutioniseffected.Dissolvethesodiumthioglycollateorthioglycolicacidinthesolutionand,ifnecessary,add1Nsodiumhydroxidesothat,aftersterilization,thesolutionwillhaveapHof7.1±0.2.Iffiltrationisnecessary,heatthesolutionagainwithoutboiling,andfilterwhilehotthroughmoistenedfilterpaper.Addtheresazurinsodiumsolution,mix,andplacethemediuminsuitablevesselsthatprovidearatioofsurfacetodepthofmediumsuchthatnotmorethantheupperhalfofthemediumhasundergoneacolorchangeindicativeofoxygenuptakeattheendoftheincubationperiod.Sterilizeusingavalidatedprocess.Ifthemediumisstored,storeatatemperaturebetween2and25inasterile,airtightcontainer.Ifmorethantheupperone-thirdofthemediumhasacquiredapinkcolor,themediummayberestoredoncebyheatingthecontainersinawater-bathorinfree-flowingsteamuntilthepinkcolordisappearsandbycoolingquickly,takingcaretopreventtheintroductionofnonsterileairintothecontainer.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净水混合，并加热至实现溶解。将巯基乙酸钠或者巯基乙酸溶解于该溶液，如果需要可再加入1N氢氧化钠，以便在灭菌后该溶液呈pH值7.1±0.2。如需要则过滤，再次加热该溶液但不得煮沸，并趁热以湿润滤纸将该溶液过滤。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，以使在培养期末表明氧气摄入的变色部分不超过培养基的上半部分。使用经过验证的工艺进行灭菌。如果需要储存该培养基，将其置于无菌、气密容器中，在2至25之间储藏。如果超过上部三分之一的培养基已经呈粉色，可以用以下方法恢复该培养基一次：在水浴锅中或者自由流动蒸气中加热该容器，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。FluidThioglycollateMediumistobeincubatedat32.5±2.5.巯基醋酸盐液体培养基将在32.5±2.5条件下进行培养。AlternativeThioglycollateMedium替代巯基醋酸盐培养基PrepareamixturehavingthesamecompositionasthatoftheFluidThioglycollateMedium,butomittingtheagarandtheresazurinsodiumsolution,sterilizeasdirectedabove,andallowtocoolpriortouse.ThepHaftersterilizationis7.1±0.2.Incubateunderanaerobicconditionsforthedurationoftheincubationperiod.配制与巯基醋酸盐液体培养基成分相同，但省略了琼脂和刃天青钠溶液的混合物，按上述方法灭菌，并在使用前静置至凉。灭菌后pH值为7.1±0.2。在厌氧条件下培养，培养时间同培养期。AlternativeFluidThioglycollateMediumistobeincubatedat32.5±2.5.替代性巯基醋酸盐培养基将在32.5±2.5条件下进行培养。Soybean–CaseinDigestMedium大豆-酪蛋白消化物培养基PancreaticDigestofCasein酪蛋白胰酶消化物17.0gPapaicDigestofSoybeanMeal大豆粉木瓜蛋白酶消化物3.0gSodiumChloride氯化钠5.0gDibasicPotassiumPhosphate磷酸氢二钾2.5gPancreaticDigestofCasein酪蛋白胰酶消化物17.0g2.5/2.3Dextrose(C6H12O6·H2O)葡萄糖gPurifiedWater纯净水1000mLDissolvethesolidsinthePurifiedWater,heatingslightlytoeffectasolution.Coolthesolutiontoroomtemperature,andadjustthepHwith1Nsodiumhydroxidesothat,aftersterilization,itwillhaveapHof7.3±0.2.Filter,ifnecessarytoclarify,dispenseintosuitablecontainers,andsterilizeusingavalidatedprocedure.Storeatatemperaturebetween2and25inasterilewell-closedcontainer,unlessitisintendedforimmediateuse.将固体物质溶解于纯净水，轻微加热以实现溶解。放凉溶液至室温，并用1N氢氧化钠调整pH值，以便在灭菌后其pH值呈7.3±0.2。过滤，如需要则使之澄清，分装入适合的容器，并用经过验证的程序消毒。如果不立刻使用，则在2到25之间以无菌且密闭良好的容器保存。Soybean–CaseinDigestMediumistobeincubatedat22.5±2.5.大豆-酪蛋白消化物培养基将在22.5±2.5条件下培养。MediaforPenicillinsorCephalosporins用于青霉素和头孢菌素的培养基WheresterilitytestmediaaretobeusedintheDirectInoculationoftheCultureMediummethodunderTestforSterilityoftheProducttobeExamined,modifythepreparationofFluidThioglycollateMediumandtheSoybean–CaseinDigestMediumasfollows.Tothecontainersofeachmedium,transferasepticallyaquantityof-lactamasesufficienttoinactivatetheamountofantibioticinthespecimenundertest.Determinethequantityof-lactamaserequiredtoinactivatetheantibioticbyusinga-lactamasepreparationthathasbeenassayedpreviouslyforitspenicillin-orcephalosporin-inactivatingpower.[NOTE—Supplemented-lactamasemediacanalsobeusedinthemembranefiltrationtest.]当无菌检查培养基用于供试产品无菌检查项下的培养基直接接种法时，按如下内容变更巯基醋酸盐液体培养基和大豆-酪蛋白消化物培养基的制备方法。向每一种培养基的容器中，以无菌操作转移足够灭活供试样品中所存在抗生素的-内酰胺酶。使用此前已经对其青霉素或头孢菌素灭活能力进行了测定的-内酰胺酶配制品，来测定灭活该抗生素所必需的-内酰胺酶数量。[注意：补充的-内酰胺酶培养基也可以用于膜过滤试验]Alternatively(inanareacompletelyseparatefromthatusedforsterilitytesting),confirmthatanappropriateamountof-lactamaseisincorporatedintothemedium,followingeithermethodunderValidationTest,usinglessthan100colony-formingunits(cfu)ofStaphylococcusaureus(seeTable1)asthechallenge.Typicalmicrobialgrowthoftheinoculatedculturemustbeobservedasaconfirmationthatthe-lactamaseconcentrationisappropriate.或者（在与无菌试验所用场所彻底隔离的区域中），按照验证试验项下的任意一种方法，使用少于100个菌落（cfu）的金黄色葡萄球菌（见表1）作为验证菌，来确认适当数量的-内酰胺酶已经被整合到该培养基中。必须观测到接种后培养物中出现典型微生物生长，才能确认-内酰胺酶浓度是适当的。Table1.StrainsoftheTestMicroorganismsSuitableforUseintheGrowthPromotionTestandtheValidationTest表1适合用于生长促进试验和验证试验中的试验微生物的菌株Aerobicbacteria好氧菌Staphylococcusaureus1ATCC6538,CIP4.83,NCTC10788,NCIMB9518金黄色葡萄球菌Bacillussubtilis枯草芽孢杆菌ATCC6633,CIP52.62,NCIMB8054Pseudomonasaeruginosa2ATCC9027,NCIMB8626,CIP82.118绿脓杆菌Anaerobicbacterium厌氧菌Clostridiumsporogenes3ATCC19404,CIP79.3,NCTC532orATCC11437产芽胞梭状芽胞杆菌Fungi霉菌Candidaalbicans白色念珠菌ATCC10231,IP48.72,NCPF3179Aspergillusniger黑曲霉ATCC16404,IP1431.83,IMI1490071AnalternativetoStaphylococcusaureusisBacillussubtilis(ATCC6633).可替代金黄色葡萄球菌的是枯草杆菌(ATCC6633)2AnalternativemicroorganismisMicrococcusluteus(Kocuriarhizophila),ATCC9341.替代微生物是藤黄微球菌（Kocuriarhizophila），ATCC9341。3AnalternativetoClostridiumsporogenes,whenanonspore-formingmicroorganismisdesired,isBacetroidesvulgatus(ATCC8482).当需要不形成芽孢微生物时，产芽胞梭状芽胞杆菌的替代微生物是Bacetroidesvulgatus[NOTE—Seed-lotculturemaintenancetechniques(seed-lotsystems)areusedsothattheviablemicroorganismsusedforinoculationarenotmorethanfivepassagesremovedfromtheoriginalmasterseedlot.][注意：采用适宜的菌种保藏技术，以确保用于接种的微生物的传代次数不超过5代]SuitabilityTests适合性试验Themediausedcomplywiththefollowingtests,carriedoutbefore,orinparallel,withthetestontheproducttobeexamined.所使用的培养基须符合下列试验，这些试验应在检验供试产品之前或者同时进行。STERILITY无菌状态Confirmthesterilityofeachsterilizedbatchofmediumbyincubatingaportionofthemediaatthespecifiedincubationtemperaturefor14days.Nogrowthofmicroorganismsoccurs.通过在指定培养温度下将一部分培养基培养14天，来确认每一批已灭菌培养基的无菌状态。不得出现微生物生长。GROWTHPROMOTIONTESTOFAEROBES,ANAEROBES,andFUNGI好氧菌、厌氧菌、霉菌的生长促进试验Testeachlotofready-preparedmediumandeachbatchofmediumpreparedeitherfromdehydratedmediumorfromingredients1.SuitablestrainsofmicroorganismsareindicatedinTable1.检查每一批已经配制好的培养基和每一批用脱水培养基或配料制备的培养基1。适当微生物菌株见表1。InoculateportionsofFluidThioglycollateMediumwithasmallnumber(notmorethan100cfu)ofthefollowingmicroorganisms,usingaseparateportionofmediumforeachofthefollowingspeciesofmicroorganism:Clostridiumsporogenes,Pseudomonasaeruginosa,andStaphylococcusaureus.InoculateportionsofAlternativeFluidThioglycollateMediumwithasmallnumber(notmorethan100cfu)ofClostridiumsporogenes.InoculateportionsofSoybean–CaseinDigestMediumwithasmallnumber(notmorethan100cfu)ofthefollowingmicroorganisms,usingaseparateportionofmediumforeachofthefollowingspeciesofmicroorganism:Aspergillusniger,Bacillussubtilis,andCandidaalbicans.Incubatefornotmorethan3daysinthecaseofbacteriaandnotmorethan5daysinthecaseoffungi.在部分巯基醋酸盐液体培养基上接种少量（不超过100cfu）下列微生物，每一种微生物均使用单独一部分培养基：产芽胞梭状芽胞杆菌、绿脓杆菌、金黄色葡萄球菌。在部分替代巯基醋酸盐液体培养基上接种少量（不超过100cfu）产芽胞梭状芽胞杆菌。在部分大豆-酪蛋白消化物培养基上接种少量（不超过100cfu）下列微生物，每一种微生物均使用单独一部分的培养基：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，霉菌培养时间不超过5天。Themediaaresuitableifaclearlyvisiblegrowthofthemicroorganismsoccurs.如果出现清晰可见的微生物生长，则该培养基是适合的。STORAGE保存Ifpreparedmediaarestoredinunsealedcontainers,theycanbeusedfor1month,providedthattheyaretestedforgrowthpromotionwithin2weeksofthetimeofuseandthatcolorindicatorrequirementsaremet.Ifstoredintightcontainers,themediacanbeusedfor1year,providedthattheyaretestedforgrowthpromotionwithin3monthsofthetimeofuseandthatthecolorindicatorrequirementsaremet.如果配制好的培养基保存于未密闭的容器中，只要在使用时间的2周内对其进行了生长促进试验并且符合颜色指示剂的要求，它们就可以使用1个月。如果保存在密闭的容器中，只要在使用时间的3个月内对其进行了生长促进试验并且符合颜色指示剂的要求，则该培养基可以使用1年。DILUTINGANDRINSINGFLUIDSFORMEMBRANEFILTRATION用于膜过滤的稀释和冲洗液FluidA液体APREPARATION配制品Dissolve1gofpepticdigestofanimaltissueinwatertomake1L,filterorcentrifugetoclarify,ifnecessary,andadjusttoapHof7.1±0.2.Dispenseintocontainers,andsterilizeusingavalidatedprocess.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，并用经过验证的工艺灭菌。PREPARATIONFORPENICILLINSORCEPHALOSPORINS用于青霉素或头孢菌素的配制品AsepticallyaddtotheabovePreparation,ifnecessary,aquantityofsterile-lactamasesufficienttoinactivateanyresidualantibioticactivityonthemembranesafterthesolutionofthetestspecimenhasbeenfiltered(seeMediaforPenicillinsorCephalosporins).在供试样品溶液已经过滤（见用于青霉素或头孢菌素的培养基）之后，如果需要，向上述配制品中，以无菌操作加入数量足够灭活滤膜上残余抗生素活性的-内酰胺酶。FluidD液体DToeachLofFluidAadd1mLofpolysorbate80,adjusttoapHof7.1±0.2,dispenseintocontainers,andsterilizeusingavalidatedprocess.Usethisfluidforarticlescontaininglecithinoroil,orfordeviceslabeledas“sterilepathway.”向每升液体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物品，或用于标为“无菌通道”的设备。FluidK液体KDissolve5.0gofpepticdigestofanimaltissue,3.0gofbeefextract,and10.0gofpolysorbate80inwatertomake1L.AdjustthepHtoobtain,aftersterilization,apHof6.9±0.2.Dispenseintocontainers,andsterilizeusingavalidatedprocess.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，以便使pH值在灭菌后呈6.9±0.2。分装入容器中，并使用经过验证的工艺灭菌。VALIDATIONTEST验证试验CarryoutatestasdescribedbelowunderTestforSterilityoftheProducttobeExaminedusingexactlythesamemethods,exceptforthefollowingmodifications.按照下面供试产品无菌检查项下的描述，使用除了下面变更之外完全相同的方法，进行试验。MembraneFiltration膜过滤Aftertransferringthecontentofthecontainerorcontainerstobetestedtothemembrane,addaninoculumofasmallnumberofviablemicroorganisms(notmorethan100cfu)tothefinalportionofsterilediluentusedtorinsethefilter.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）试验菌.DirectInoculation直接接种Aftertransferringthecontentsofthecontainerorcontainerstobetested(forcatgutandothersurgicalsuturesforveterinaryuse:strands)totheculturemedium,addaninoculumofasmallnumberofviablemicroorganisms(notmorethan100cfu)tothemedium.在将一个或多个供试容器（对于兽医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。InbothcasesusethesamemicroorganismsasthosedescribedaboveunderGrowthPromotionTestofAerobes,Anaerobes,andFungi.Performagrowthpromotiontestasapositivecontrol.Incubateallthecontainerscontainingmediumfornotmorethan5days.在这两种情况中，均按照上述好氧菌、厌氧菌、霉菌生长促进试验项下的描述，使用同样的微生物。进行一个生长促进试验作为阳性对照。培养所有含有培养基的容器，培养时间不超过5天。Ifclearlyvisiblegrowthofmicroorganismsisobtainedaftertheincubation,visuallycomparabletothatinthecontrolvesselwithoutproduct,eithertheproductpossessesnoantimicrobialactivityundertheconditionsofthetestorsuchactivityhasbeensatisfactorilyeliminated.Thetestforsterilitymaythenbecarriedoutwithoutfurthermodification.如果在培养后得到清晰可见的微生物生长，看起来与没有产品的对照容器中的生长类似，则该产品在此试验条件下没有任何抗微生物活性，或者此活性已经被令人满意地消除了。然后，无菌试验可以进行，而无需进一步的变更。Ifclearlyvisiblegrowthisnotobtainedinthepresenceoftheproducttobetested,visuallycomparabletothatinthecontrolvesselswithoutproduct,theproductpossessesantimicrobialactivitythathasnotbeensatisfactorilyeliminatedundertheconditionsofthetest.Modifytheconditionsinordertoeliminatetheantimicrobialactivity,andrepeatthevalidationtest.如果用肉眼与没有产品的对照容器比较，无法在存在供试产品的情况下得到清晰可见的生长，则该产品在试验条件下所具有的抗微生物活性尚未令人满意地消除。变更条件以便消除抗微生物活性，并重复验证试验。Thisvalidationisperformed(a)whenthetestforsterilityhastobecarriedoutonanewproduct;and(b)wheneverthereisachangeintheexperimentalconditionsofthetest.ThevalidationmaybeperformedsimultaneouslywiththeTestforSterilityoftheProducttobeExamined.当(a)一个新产品必须进行无菌试验时，和(b)无论何时该试验的试验条件发生改变时，则需进行此验证试验。该验证可以与供试产品的无菌试验同时进行。TESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINED供试产品的无菌检查NumberofArticlestoBeTested供试物品数量Unlessotherwisespecifiedelsewhereinthischapterorintheindividualmonograph,testthenumberofarticlesspecifiedinTable3.Ifthecontentsofeacharticleareofsufficientquantity(seeTable2),theymaybedividedsothatequalappropriateportionsareaddedtoeachofthespecifiedmedia.[NOTE—Performsterilitytestingemployingtwoormoreofthespecifiedmedia.]Ifeacharticledoesnotcontainsufficientquantitiesforeachmedium,usetwicethenumberofarticlesindicatedinTable3.除非在此章节的其他位置或在具体的各论中另有规定，供试物品的数量遵照表3中的规定。如果每个物品的内容物有足够数量（见表2），可以将其分成若干等份，将适当的等份加入到每个指定的培养基。[注意：使用两个或更多指定培养基，来进行无菌试验。]如果每个物品内容物的数量不够每个培养基的用量，使用表3中所规定物品数量的2倍。Table2.MinimumQuantitytobeUsedforEachMedium表2：用于每个培养基的最小数量MinimumQuantitytobeUsed(unlessotherwisejustifiedandauthorized)最小使用数量(除非另有依据和授权)QuantityperContainer每个容器中的数量Liquids(otherthananitbiotics)液体(除了抗生素)Lessthan1mLThewholecontentsofeachcontainer少于1mL每个容器的总内容物1–40mLHalfthecontentsofeachcontainer,butnotlessthan1mL每个容器中内容物的一半，但不得少于1mLGreaterthan40mL,andnotgreaterthan100m20mLL大于40mL，但不大于100mLGreaterthan100mL10%ofthecontentsofthecontainer,大于100mLbutnotlessthan20mL该容器内容物的10%，但不得少于20mLAntibioticliquids1mL抗生素液体MinimumQuantitytobeUsed(unlessotherwisejustifiedandauthorized)最小使用数量(除非另有依据和授权)QuantityperContainer每个容器中的数量OtherpreparationssolubleinwaterorinisThewholecontentsofeachcontaineropropylmyristatetoprovidenotlessthan200mg溶于水或豆蔻酸异丙酯的其他配制品每个容器的全部内容物，以提供不少于200mgInsolublepreparations,creams,andointmUsethecontentsofeachcontainertoentstobesuspendedoremulsifiedprovidenotlessthan200mg待悬浮或乳化的不溶性配制品、乳膏、油使用每个容器的内容物，以提供不膏少于200mgSolids固体Lessthan50mgThewholecontentsofeachcontainer少于50mg每个容器的全部内容物50mgormore,butlessthan300mgHalfthecontentsofeachcontainer,50mg或者更多，但少于300mgbutnotlessthan50mg每个容器内容物的一半，但不少于50mgMinimumQuantitytobeUsed(unlessotherwisejustifiedandauthorized)最小使用数量(除非另有依据和授权)QuantityperContainer每个容器中的数量300mg–5g150mgGreaterthan5g多于5g500mgDevices设备Catgutandothersurgicalsuturesforveter3sectionsofastrand(each30-inaryusecmlong)兽医用肠线和其他外科缝合线一股线的3部分(每个30cm长)Surgicaldressing/cotton/gauze(inpacka100mgperpackage100mg每包装ges)外科敷料/棉花/纱布(在包装中)SuturesandotherindividuallypackagedsiThewholedevice整个设备ngle-usematerial缝合线合其他单个包装的一次性使用物料OthermedicaldevicesThewholedevice,cutintopiecesord其他医用设备isassembled整个设备，切成片或拆开Table3.MinimumNumberofArticlestobeTestedinRelationtotheNumberofArticlesintheBatch表3：与物品批量相关的最小供试物品数量MinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)*每个培养基中的最小供试物品数量NumberofItemsintheBatch该批物品的数量(除非另有依据或授权)Parenteralpreparations注射用药的配制品Notmorethan100containers10%or4containers,whicheveristh不多于100个容器egreater10%或4个容器，选较多者Morethan100butnotmorethan500containers10containers多于100个但不多于500个容器10个容器Morethan500containers2%or20containers,whicheverisle多于500个容器ss2%或者20容器，选较少者Forlarge-volumeparenterals2%or10containers,whicheverisle对于大体积注射用药制剂ss2%或者10容器，选较少者MinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)*每个培养基中的最小供试物品数量NumberofItemsintheBatch该批物品的数量(除非另有依据或授权)Antibioticsolids固体抗生物Pharmacybulkpackages(<5g)20containers药房散装(<5g)20个容器Pharmacybulkpackages(5g)6containers药房散装(5g)6个容器BulksandblendsSeeBulksolidproducts散装并混合见散装固体产品Ophthalmicandothernoninjectablepreparations眼科和其他非注射配制品Notmorethan200containers5%or2containers,whicheveristhe不多于200个容器greater5%或者2个容器，选较多者Morethan200containers10containersMinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)*每个培养基中的最小供试物品数量NumberofItemsintheBatch该批物品的数量(除非另有依据或授权)多于200个容器10个容器Iftheproductispresentedintheformofsingle-dosecontainers,applytheschemeshownaboveforpreparationsforparenteraluse.如果该产品存在于单一剂量容器中，应用上述用于注射用药配制品的方案Devices设备Catgutandothersurgicalsuturesforveteri2%or5packages,whicheveristhegrnaryuseeater,兽医用肠线和其他外科缝合线uptoamaximumtotalof20packages2%或5个包装，选较多者，最多可达20个包装Notmorethan100articles10%or4articles,whicheverisgrea不多于100个物品terMinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)*每个培养基中的最小供试物品数量NumberofItemsintheBatch该批物品的数量(除非另有依据或授权)10%或4个物品，选较多者Morethan100,butnotmorethan500articles10articles多于100但不多于500个物品10个物品Morethan500articles2%or20articles,whicheverisless多于500个物品2%或20个物品，选较少者Bulksolidproducts散装固体产品Upto4containersEachcontainer最多4个容器每个容器Morethan4containers,butnotmorethan50co20%or4containers,whicheverisgrntainerseater多于4个容器，但不多于50个容器20%或4个容器，选较多者Morethan50containers2%or10containers,whicheverisgr超过50个容器eater2%或者10个容器，选较多者MinimumNumberofItemstobeTestedforEachMedium(unlessotherwisejustifiedandauthorized)*每个培养基中的最小供试物品数量NumberofItemsintheBatch该批物品的数量(除非另有依据或授权)*Ifthecontentsofonecontainerareenoughtoinoculatethetwomedia,thiscolumngivesthenumberofcontainersneededforboththemediatogether.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。ThetestmaybecarriedoutusingthetechniqueofMembraneFiltrationorbyDirectInoculationoftheCultureMediumwiththeproducttobeexamined.Appropriatenegativecontrolsareincluded.Thetechniqueofmembranefiltrationisusedwheneverthenatureoftheproductpermits;thatis,forfilterableaqueouspreparations,foralcoholicoroilypreparations,andforpreparationsmisciblewith,orsolublein,aqueousoroilysolvents,providedthesesolventsdonothaveanantimicrobialeffectintheconditionsofthetest.此试验可以使用膜过滤法或培养基直接接种法进行。应包括多个适当的阴性对照。只要该产品的性质许可，就应使用膜过滤法；这些性质是，可过滤的水溶性配制品、酒精或油性配制品、易混合或溶解于水或油性溶剂的配制品，只要这些溶剂在试验条件下没有抗生素效果。MembraneFiltration膜过滤Usemembranefiltershavinganominalporesizenotgreaterthan0.45µmwhoseeffectivenesstoretainmicroorganismshasbeenestablished.Cellulosenitratefilters,forexample,areusedforaqueous,oily,andweaklyalcoholicsolutions;andcelluloseacetatefilters,forexample,areusedforstronglyalcoholicsolutions.Speciallyadaptedfiltersmaybeneededforcertainproducts(e.g.,forantibiotics).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过滤器。Thetechniquedescribedbelowassumesthatmembranesabout50mmindiameterwillbeused.Iffiltersofadifferentdiameterareused,thevolumesofthedilutionsandthewashingsshouldbeadjustedaccordingly.Thefiltrationapparatusandmembranearesterilizedbyappropriatemeans.Theapparatusisdesignedsothatthesolutiontobeexaminedcanbeintroducedandfilteredunderasepticconditions:itpermitstheasepticremovalofthemembranefortransfertothemedium,oritissuitableforcarryingouttheincubationafteraddingthemediumtotheapparatusitself.下面描述的方法所使用直径约50mm的滤膜。如果使用不同直径的过滤器，稀释液和洗液的体积应当作相应调节。以适当方法将过滤设备和滤膜灭菌。该设备设计用于在无菌条件下加入和过滤供试溶液：其使得在无菌状态下将滤膜摘掉转移至培养基成为可能，或者其适合于将培养基加入该设备自身之中，并进行培养。AQUEOUSSOLUTIONS水性溶液Ifappropriate,transferasmallquantityofasuitable,sterilediluentsuchasFluidA(seeDilutingandRinsingFluidsforMembraneFiltration)ontothemembraneintheapparatusandfilter.Thediluentmaycontainsuitableneutralizingsubstancesand/orappropriateinactivatingsubstances,forexample,inthecaseofantibiotics.如果适当，将少量适当的无菌稀释剂，例如液体A（见用于膜过滤的稀释和冲洗液），转移至设备中的滤膜上并过滤。该稀释剂可以含有适当的中和物质和/或适当的灭活物质，例如针对抗生素。Transferthecontentsofthecontainerorcontainerstobetestedtothemembraneormembranes,ifnecessary,afterdilutingtothevolumeusedintheValidationTestwiththechosensterilediluent,butusingnotlessthanthequantitiesoftheproducttobeexaminedprescribedinTables2and3.Filterimmediately.Iftheproducthasantimicrobialproperties,washthemembranenotlessthanthreetimesbyfilteringthroughiteachtimethevolumeofthechosensterilediluentusedintheValidationTest.Donotexceedawashingcycleof5times200mL,evenifduringvalidationithasbeendemonstratedthatsuchacycledoesnotfullyeliminatetheantimicrobialactivity.Transferthewholemembranetotheculturemediumorcutitasepticallyintotwoequalparts,andtransferonehalftoeachoftwosuitablemedia.UsethesamevolumeofeachmediumasintheValidationTest.Alternatively,transferthemediumontothemembraneintheapparatus.Incubatethemediafornotlessthan14days.将一个或多个供试容器的内容物转移到滤膜，如需要可先使用选定的无菌稀释剂稀释至验证试验中所用体积，但须使用不少于表2和3中规定的供试产品数量。立即过滤。如果该产品具有抗微生物特性，冲洗滤膜不少于3次，每次均将验证试验中所使用的无菌稀释剂体积滤过该滤膜。即便验证中显示5次200mL的冲洗循环没有完全消除抗微生物活性，也不要超越这样一个循环。转移整个滤膜至培养基，或以无菌操作将其切开至相等的2部分，并将每一部分转移至适当的培养基中。每个培养基的体积与验证试验所用的一样。或者，将培养基转移至设备中的滤膜上。培养该培养基，不少于14天。SOLUBLESOLIDS(otherthanantibiotics)可溶固体（非抗生素）UseforeachmediumnotlessthanthequantityprescribedinTables2and3oftheproductdissolvedinasuitablesolvent,suchasFluidA(DilutingandRinsingFluidsforMembraneFiltration),andproceedwiththetestasdescribedaboveforAqueousSolutionsusingamembraneappropriatetothechosensolvent.在每个培养基中，使用不少于表2和3规定的产品数量溶于适当溶剂，例如溶液A（用于膜过滤的稀释和冲洗液），并按照上述关于水性溶液的描述，使用适合所选溶剂的滤膜，继续进行该试验。OILSandOILYSOLUTIONS油和油性溶液UseforeachmediumnotlessthanthequantityoftheproductprescribedinTables2and3.Oilsandoilysolutionsofsufficientlylowviscositymaybefilteredwithoutdilutionthroughadrymembrane.Viscousoilsmaybedilutedasnecessarywithasuitablesterilediluentsuchasisopropylmyristateshownnottohaveantimicrobialactivityintheconditionsofthetest.Allowtheoiltopenetratethemembranebyitsownweight,andthenfilter,applyingthepressureorsuctiongradually.Washthemembraneatleastthreetimesbyfilteringthroughiteachtimeabout100mLofasuitablesterilesolutionsuchasFluidA(seeDilutingandRinsingFluidsforMembraneFiltration)containingasuitableemulsifyingagentataconcentrationshowntobeappropriateinthevalidationofthetest,forexamplepolysorbate80ataconcentrationof10gperL(FluidK).Transferthemembraneormembranestotheculturemediumormedia,orviceversa,asdescribedaboveforAqueousSolutions,andincubateatthesametemperaturesandforthesametimes.在每个培养基中，使用不少于表2和3中描述的产品用量。粘性足够低的油和油性溶液可能在不经稀释的情况下滤过干燥滤膜。如需要，粘稠油质可以用适合的无菌稀释剂进行稀释，例如已证实在该试验条件下不具有抗微生物活性的豆蔻酸异丙酯。使该油质依靠其自身的重量穿过滤膜，然后逐渐应用压力或抽吸过滤。每次过滤约100mL适当的无菌溶液，例如液体A（见用于膜过滤的稀释和冲洗液），并含有适当乳化剂且其浓度已证实适用于该试验的验证，例如浓度为10克每升的聚山梨酯80（液体K）。将一个或多个滤膜转移到一个或多个培养基，或反之，如上面关于水性溶液的描述，并在相同温度下培养同样的时间。OINTMENTSandCREAMS油膏和乳膏UseforeachmediumnotlessthanthequantitiesoftheproductprescribedinTables2and3.Ointmentsinafattybaseandemulsionsofthewater-in-oiltypemaybedilutedto1%inisopropylmyristateasdescribedabove,byheating,ifnecessary,tonotmorethan40.Inexceptionalcasesitmaybenecessarytoheattonotmorethan44.Filterasrapidlyaspossible,andproceedasdescribedaboveforOilsandOilySolutions.在每个培养基中，使用不少于表2和3中描述的产品用量。脂肪状的油膏和水在油中形态乳化剂可以按照上面所述，在豆蔻酸异丙酯中稀释至1%，如需要可加热至不高于40。在特别情况下，其可能必须加热到不超过44。尽可能迅速地过滤，并按照上面针对油和油性溶液所述内容继续操作。PREFILLEDSYRINGES预装填的注射器Forprefilledsyringeswithoutattachedsterileneedles,expelthecontentsofeachsyringeintooneortwoseparatemembranefilterfunnelsorintoseparatepoolingvesselspriortotransfer.Ifaseparatesterileneedleisattached,directlyexpelthesyringecontentsasindicatedabove,andproceedasdirectedforAqueousSolutions.Testthesterilityoftheneedle,usingDirectInoculationunderValidationTest.对于没有附无菌针头的预装填注射器，在转移之前，将每个注射器的内容物排出至一个或两个单独的膜过滤器漏斗，或至若干单独的合并容器。如果附了单独的灭菌针头，直接按照上面的规定将注射器内容物直接排出，并按照关于水性溶液的规定继续进行。使用验证试验项下的直接接种，检查针头的无菌情况。SOLIDSFORINJECTIONOTHERTHANANTIBIOTICS除了抗生素之外的注射用固体Constitutethetestarticlesasdirectedonthelabel,andproceedasdirectedforAqueousSolutionsorOilsandOilySolutions,whicheverapplies.[NOTE—Ifnecessary,excessdiluentcanbeaddedtoaidintheconstitutionandfiltrationoftheconstitutedtestarticle.]按照其标签上的规定配制供试物品，并按照适用的关于水性溶液或油和油性溶液的规定继续进行。[注意：如需要，可以加入额外的稀释剂以帮助对已配制的供试物品进行再配制和过滤]ANTIBIOTICSOLIDSFORINJECTION用于注射的抗生素PharmacyBulkPackages,<5g—Fromeachof20containers,asepticallytransferabout300mgofsolids,intoasterile500-mLconicalflask,dissolveinabout200mLofFluidA(seeDilutingandRinsingFluidsforMembraneFiltration),andmix;orconstitute,asdirectedinthelabeling,eachof20containersandtransferaquantityofliquidorsuspension,equivalenttoabout300mgofsolids,intoasterile500-mLconicalflask,dissolveinabout200mLofFluidA,andmix.ProceedasdirectedforAqueousSolutionsorOilsandOilySolutions,whicheverapplies.药房散装<5g：取20个容器，每个容器均以无菌操作将约300mg固体转移至一个无菌的500mL锥形烧瓶，溶解于约200mL液体A中（见用于膜过滤的稀释和冲洗液），并混匀；或取20个容器，每个容器均按照标签上的规定配制，并将相当于大约300mg固体的液体或悬浮液转移至一个无菌的500mL锥形烧瓶，溶解于约200mL液体A中，并混匀。按照适合的关于水性溶液或油和油性溶液的规定，继续操作。PharmacyBulkPackages,5g—Fromeachof6containers,asepticallytransferabout1gofsolidsintoasterile500-mLconicalflask,dissolveinabout200mLofFluidA,andmix;orconstitute,asdirectedinthelabeling,eachof6containersandtransferaquantityofliquid,equivalenttoabout1gofsolids,intoasterile500-mLconicalflask,dissolveinabout200mLofFluidA,andmix.ProceedasdirectedforAqueousSolutions.药方散装5g：取6个容器，每个均以无菌操作将大约1克的固体转移至一个无菌的500mL锥形烧瓶，溶解于约200mL液体A中，并混匀；或取6个容器，每个均按照标签的规定配制，将相当于1g固体的液体转移至一个无菌的500mL锥形烧瓶，溶解于200mL液体A中，并混匀。