美国药典USP31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1211MEDIA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2.5gDextrose(C6H12O6·H2O)葡萄糖5.5/5.0gAgar,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)5.0gPancreaticDigestofCasein15.0gL-CystineL-胱氨酸0.5g酪蛋白胰酶消化物SodiumThioglycollate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