無菌檢查方案

71STERILITYTESTS無菌測試PORTIONSOFTHISGENERALCHAPTERHAVEBEENHARMONIZEDWITHTHECORRESPONDINGTEXTSOFTHEEUROPEANPHARMACOPEIAAND/ORTHEJAPANESEPHARMACOPEIATHOSEPORTIONSTHATARENOTHARMONIZEDAREMARKEDWITHSYMBOLSTOSPECIFYTHISFACT此章節(jié)的某些部份與歐洲藥典和/或日本藥典的相關(guān)內(nèi)容一致。不一致的章節(jié)用（）符號來表示，以標(biāo)明其事實(shí)。THEFOLLOWINGPROCEDURESAREAPPLICABLEFORDETERMININGWHETHERAPHARMACOPEIALARTICLEPURPORTINGTOBESTERILECOMPLIESWITHTHEREQUIREMENTSSETFORTHINTHEINDIVIDUALMONOGRAPHWITHRESPECTTOTHETESTFORSTERILITYPHARMACOPEIALARTICLESARETOBETESTEDBYTHEMEMBRANEFILTRATIONMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDWHERETHENATUREOFTHEPRODUCTPERMITSIFTHEMEMBRANEFILTRATIONTECHNIQUEISUNSUITABLE,USETHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDALLDEVICES,WITHTHEEXCEPTIONOFDEVICESWITHPATHWAYSLABELEDSTERILE,ARETESTEDUSINGTHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODPROVISIONSFORRETESTINGAREINCLUDEDUNDEROBSERVATIONANDINTERPRETATIONOFRESULTS以下程序用來確認(rèn)藥典藥品是否按單獨(dú)記錄所列相關(guān)無菌測試要求執(zhí)行。藥品的特性允許下，按受檢藥品無菌測試中的薄膜過濾法來測試藥典藥品。如果薄膜過濾法不適用，用受檢藥品無菌測試中的培養(yǎng)基的直接接種指示方法來測試。除設(shè)備路徑標(biāo)明無菌外，所有的儀器全用培養(yǎng)基的直接接種指示來測試。測試的物品包含于結(jié)果觀察資料及說明。BECAUSESTERILITYTESTINGISAVERYEXACTINGPROCEDURE,WHEREASEPSISOFTHEPROCEDUREMUSTBEENSUREDFORACORRECTINTERPRETATIONOFRESULTS,ITISIMPORTANTTHATPERSONNELBEPROPERLYTRAINEDANDQUALIFIEDTHETESTFORSTERILITYISCARRIEDOUTUNDERASEPTICCONDITIONSINORDERTOACHIEVESUCHCONDITIONS,THETESTENVIRONMENTHASTOBEADAPTEDTOTHEWAYINWHICHTHESTERILITYTESTISPERFORMEDTHEPRECAUTIONSTAKENTOAVOIDCONTAMINATIONARESUCHTHATTHEYDONOTAFFECTANYMICROORGANISMSTHATARETOBEREVEALEDINTHETESTTHEWORKINGCONDITIONSINWHICHTHETESTSAREPERFORMEDAREMONITOREDREGULARLYBYAPPROPRIATESAMPLINGOFTHEWORKINGAREAANDBYCARRYINGOUTAPPROPRIATECONTROLS因?yàn)闊o菌測試是一個極其嚴(yán)格的測試，必需確保滅菌程序以求結(jié)果的正確詮釋，所以對人員進(jìn)行相應(yīng)培訓(xùn)以至合格就顯得極為重要。無菌測試在無菌條件下進(jìn)行。為了達(dá)到這樣的環(huán)境，應(yīng)改變周圍的測試環(huán)境至無菌測試環(huán)境。應(yīng)采取預(yù)防措施避免污染，并且此措施不影響測試中有機(jī)體的暴露。無菌測試的工作環(huán)境應(yīng)靠在工作區(qū)取樣來接受定期監(jiān)控，同時進(jìn)行適當(dāng)控制。THESEPHARMACOPEIALPROCEDURESARENOTBYTHEMSELVESDESIGNEDTOENSURETHATABATCHOFPRODUCTISSTERILEORHASBEENSTERILIZEDTHISISACCOMPLISHEDPRIMARILYBYVALIDATIONOFTHESTERILIZATIONPROCESSOROFTHEASEPTICPROCESSINGPROCEDURES這些醫(yī)典中的程序并非由自己確定一批藥品的無菌性，或者是已經(jīng)過滅菌的。它主要是由滅菌過程和無菌加工程序認(rèn)證來確定的。WHENEVIDENCEOFMICROBIALCONTAMINATIONINTHEARTICLEISOBTAINEDBYTHEAPPROPRIATEPHARMACOPEIALMETHOD,THERESULTSOOBTAINEDISCONCLUSIVEEVIDENCEOFFAILUREOFTHEARTICLETOMEETTHEREQUIREMENTSOFTHETESTFORSTERILITY,EVENIFADIFFERENTRESULTISOBTAINEDBYANALTERNATIVEPROCEDUREFORADDITIONALINFORMATIONONSTERILITYTESTING,SEESTERILIZATIONANDSTERILITYASSURANCEOFCOMPENDIALARTICLES1211當(dāng)通過相應(yīng)藥典中的方法得到的藥品中的微生物污染證據(jù)，即使另一程序得到不同結(jié)果，以此得到的結(jié)果為藥品不符合無菌測試要求的最終結(jié)果。無菌測試的詳細(xì)信息見COMPENDIALARTICLES的滅菌和無菌保證1211MEDIA培養(yǎng)基PREPAREMEDIAFORTHETESTSASDESCRIBEDBELOW,ORDEHYDRATEDFORMULATIONSMAYBEUSEDPROVIDEDTHAT,WHENRECONSTITUTEDASDIRECTEDBYTHEMANUFACTURERORDISTRIBUTOR,THEYMEETTHEREQUIREMENTSOFTHEGROWTHPROMOTIONTESTOFAEROBES,ANAEROBES,ANDFUNGIMEDIAARESTERILIZEDUSINGAVALIDATEDPROCESS按以下所述為測試準(zhǔn)備培養(yǎng)基，或無水配方，當(dāng)按生產(chǎn)商和銷售商指示重新組成，它們符合需氧菌，厭氧性生物，和真菌類助長測試要求。按驗(yàn)證方法對培養(yǎng)基進(jìn)行滅菌。THEFOLLOWINGCULTUREMEDIAHAVEBEENFOUNDTOBESUITABLEFORTHETESTFORSTERILITYFLUIDTHIOGLYCOLLATEMEDIUMISPRIMARILYINTENDEDFORTHECULTUREOFANAEROBICBACTERIAHOWEVER,ITWILLALSODETECTAEROBICBACTERIASOYBEANCASEINDIGESTMEDIUMISSUITABLEFORTHECULTUREOFBOTHFUNGIANDAEROBICBACTERIA以下培養(yǎng)基適用于無菌測試。液態(tài)硫乙醇酸鹽培養(yǎng)基主要是用來培養(yǎng)厭氧菌。大豆豆蛋白消化物培養(yǎng)基適用于培養(yǎng)真菌和需氧菌。FLUIDTHIOGLYCOLLATEMEDIUM液態(tài)硫乙醇酸鹽培養(yǎng)基LCYSTINEL胱氨酸05GSODIUMCHLORIDE氯化鈉25GDEXTROSEC6H12O6H2O葡萄糖55/50GAGAR,GRANULATEDMOISTURECONTENTNOTEXCEEDING15瓊脂，顆粒狀（濕度不超過15）075GYEASTEXTRACTWATERSOLUBLE酵母浸出粉（水溶性）50GPANCREATICDIGESTOFCASEIN酪蛋白胰酶消化物150GSODIUMTHIOGLYCOLLATE硫乙醇酸鈉05GORTHIOGLYCOLICACID或者疏乙醋酸03MLRESAZURINSODIUMSOLUTION1IN1000,FRESHLYPREPARED新配制的（1/1000）刃天青鈉溶液10MLPURIFIEDWATER水1000MLMIXTHELCYSTINE,SODIUMCHLORIDE,DEXTROSE,YEASTEXTRACT,ANDPANCREATICDIGESTOFCASEINWITHTHEPURIFIEDWATER,ANDHEATUNTILSOLUTIONISEFFECTEDDISSOLVETHESODIUMTHIOGLYCOLLATEORTHIOGLYCOLICACIDINTHESOLUTIONAND,IFNECESSARY,ADD1NSODIUMHYDROXIDESOTHAT,AFTERSTERILIZATION,THESOLUTIONWILLHAVEAPHOF7102IFFILTRATIONISNECESSARY,HEATTHESOLUTIONAGAINWITHOUTBOILING,ANDFILTERWHILEHOTTHROUGHMOISTENEDFILTERPAPERADDTHERESAZURINSODIUMSOLUTION,MIX,ANDPLACETHEMEDIUMINSUITABLEVESSELSTHATPROVIDEARATIOOFSURFACETODEPTHOFMEDIUMSUCHTHATNOTMORETHANTHEUPPERHALFOFTHEMEDIUMHASUNDERGONEACOLORCHANGEINDICATIVEOFOXYGENUPTAKEATTHEENDOFTHEINCUBATIONPERIODSTERILIZEUSINGAVALIDATEDPROCESSIFTHEMEDIUMISSTORED,STOREATATEMPERATUREBETWEEN2AND25INASTERILE,AIRTIGHTCONTAINERIFMORETHANTHEUPPERONETHIRDOFTHEMEDIUMHASACQUIREDAPINKCOLOR,THEMEDIUMMAYBERESTOREDONCEBYHEATINGTHECONTAINERSINAWATERBATHORINFREEFLOWINGSTEAMUNTILTHEPINKCOLORDISAPPEARSANDBYCOOLINGQUICKLY,TAKINGCARETOPREVENTTHEINTRODUCTIONOFNONSTERILEAIRINTOTHECONTAINERFLUIDTHIOGLYCOLLATEMEDIUMISTOBEINCUBATEDAT32525用純靜水將L胱氨酸，氯化鈉，葡萄糖，酵母浸出粉，酪蛋白胰酶消化物混合。加熱直至形成溶液，在溶液中溶解硫乙醇酸鈉或疏乙醋酸，必要時，加入1N的氫氧化鈉，在滅菌后，PH值會達(dá)到7102。如果有必要過濾的話，加熱溶液但不用達(dá)到沸點(diǎn)，熱氣通過潮濕的過濾紙時進(jìn)行過濾。加入刃天青鈉溶液，并攪拌混合，將培養(yǎng)基放入適當(dāng)?shù)娜萜髦?，此容器可以顯示一個培養(yǎng)基表面至深度的比率，培育階段末時，不超過培養(yǎng)基上半部份的指示劑氧化層發(fā)生顏色變化。用驗(yàn)證方法進(jìn)行滅菌。如果要存儲培養(yǎng)基，將它存儲于一個2和25間的無菌密封容器中。如果超過三分之一的培養(yǎng)基顯示粉紅色，在水浴或自由流通蒸汽中加熱容器直至粉紅色消失，并快速冷卻，并防止有菌氣體產(chǎn)生并進(jìn)入容器，液態(tài)硫乙醇酸鹽培養(yǎng)基就在32525下進(jìn)行培育。ALTERNATIVETHIOGLYCOLLATEMEDIUM可選擇疏基醋酸液培養(yǎng)基PREPAREAMIXTUREHAVINGTHESAMECOMPOSITIONASTHATOFTHEFLUIDTHIOGLYCOLLATEMEDIUM,BUTOMITTINGTHEAGARANDTHERESAZURINSODIUMSOLUTION,STERILIZEASDIRECTEDABOVE,ANDALLOWTOCOOLPRIORTOUSETHEPHAFTERSTERILIZATIONIS7102INCUBATEUNDERANAEROBICCONDITIONSFORTHEDURATIONOFTHEINCUBATIONPERIOD按液態(tài)硫乙醇酸鹽培養(yǎng)基所含成份同樣準(zhǔn)備一份混合物，除去瓊脂和刃天青鈉溶液。按以上所示滅菌，使用前允許進(jìn)行冷卻。滅菌后PH值為7102。在厭氧條件下進(jìn)行培育，以求培育期的持續(xù)。ALTERNATIVEFLUIDTHIOGLYCOLLATEMEDIUMISTOBEINCUBATEDAT32525可選擇疏基醋酸液培養(yǎng)基應(yīng)在32525下進(jìn)行培育。SOYBEANCASEINDIGESTMEDIUM大豆酪蛋白消化物培養(yǎng)基PANCREATICDIGESTOFCASEIN酪蛋白胰酶消化物170GPAPAICDIGESTOFSOYBEANMEAL大豆粉木瓜蛋白酶水化物30GSODIUMCHLORIDE氯化鈉50GDIBASICPOTASSIUMPHOSPHATE磷酸氫二鉀25GDEXTROSEC6H12O6H2O葡萄糖25/23GPURIFIEDWATER水1000MLDISSOLVETHESOLIDSINTHEPURIFIEDWATER,HEATINGSLIGHTLYTOEFFECTASOLUTIONCOOLTHESOLUTIONTOROOMTEMPERATURE,ANDADJUSTTHEPHWITH1NSODIUMHYDROXIDESOTHAT,AFTERSTERILIZATION,ITWILLHAVEAPHOF7302FILTER,IFNECESSARYTOCLARIFY,DISPENSEINTOSUITABLECONTAINERS,ANDSTERILIZEUSINGAVALIDATEDPROCEDURESTOREATATEMPERATUREBETWEEN2AND25INASTERILEWELLCLOSEDCONTAINER,UNLESSITISINTENDEDFORIMMEDIATEUSE在水中溶解固體，輕輕加熱以形成溶液。將溶液冷卻至室溫，用1N的氫氧化鈉來調(diào)節(jié)PH值，以至在滅菌后其PH值達(dá)到7302。過濾，如果有必要進(jìn)行澄清時，將漏斗放入相應(yīng)的容器中，并用驗(yàn)證方法來進(jìn)行滅菌。在一個無菌密封容器中2和25下進(jìn)行存儲，直至使用。SOYBEANCASEINDIGESTMEDIUMISTOBEINCUBATEDAT22525大豆酪蛋白消化物培養(yǎng)基在22525下進(jìn)行培育。MEDIAFORPENICILLINSORCEPHALOSPORINS青霉素或頭孢菌素培養(yǎng)基WHERESTERILITYTESTMEDIAARETOBEUSEDINTHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINED,MODIFYTHEPREPARATIONOFFLUIDTHIOGLYCOLLATEMEDIUMANDTHESOYBEANCASEINDIGESTMEDIUMASFOLLOWSTOTHECONTAINERSOFEACHMEDIUM,TRANSFERASEPTICALLYAQUANTITYOFLACTAMASESUFFICIENTTOINACTIVATETHEAMOUNTOFANTIBIOTICINTHESPECIMENUNDERTESTDETERMINETHEQUANTITYOFLACTAMASEREQUIREDTOINACTIVATETHEANTIBIOTICBYUSINGALACTAMASEPREPARATIONTHATHASBEENASSAYEDPREVIOUSLYFORITSPENICILLINORCEPHALOSPORININACTIVATINGPOWERNOTESUPPLEMENTEDLACTAMASEMEDIACANALSOBEUSEDINTHEMEMBRANEFILTRATIONTEST當(dāng)用受檢藥品無菌試驗(yàn)測試的培養(yǎng)基直接接種方法來進(jìn)行無菌試驗(yàn)時，按以下更正液態(tài)硫乙醇酸鹽培養(yǎng)基和大豆酪蛋白消化物培養(yǎng)基的制劑。在無菌情況下，將足夠的內(nèi)酰胺酶轉(zhuǎn)移到每種培養(yǎng)基的容器中，來鈍化試驗(yàn)樣品中的抗生素。用以前化驗(yàn)青霉素或頭孢菌素鈍化力的內(nèi)酰胺酶制劑來確定需要鈍化抗生素的內(nèi)酰胺酶的量。ALTERNATIVELYINANAREACOMPLETELYSEPARATEFROMTHATUSEDFORSTERILITYTESTING,CONFIRMTHATANAPPROPRIATEAMOUNTOFLACTAMASEISINCORPORATEDINTOTHEMEDIUM,FOLLOWINGEITHERMETHODUNDERVALIDATIONTEST,USINGLESSTHAN100COLONYFORMINGUNITSCFUOFSTAPHYLOCOCCUSAUREUSSEETABLE1ASTHECHALLENGETYPICALMICROBIALGROWTHOFTHEINOCULATEDCULTUREMUSTBEOBSERVEDASACONFIRMATIONTHATTHELACTAMASECONCENTRATIONISAPPROPRIATE另外（在另一個完全與無菌試驗(yàn)無關(guān)的領(lǐng)域），確定適當(dāng)量內(nèi)酰胺酶混合于培養(yǎng)基中，接著用驗(yàn)證測試中的方法，用不少于100CFU葡萄狀球菌（見表1）做為挑戰(zhàn)性實(shí)驗(yàn)接種的培養(yǎng)菌的典型微生物生長應(yīng)接受觀察，以確定內(nèi)酰胺酶的濃渡適當(dāng)。TABLE1STRAINSOFTHETESTMICROORGANISMSSUITABLEFORUSEINTHEGROWTHPROMOTIONTESTANDTHEVALIDATIONTEST表1。促生長試驗(yàn)和驗(yàn)證試驗(yàn)中的可用到微生物菌株AEROBICBACTERIA需氧細(xì)菌STAPHYLOCOCCUSAUREUS1金黃色葡萄球菌ATCC6538,CIP483,NCTC10788,NCIMB9518BACILLUSSUBTILIS枯草芽孢桿菌ATCC6633,CIP5262,NCIMB8054PSEUDOMONASAERUGINOSA2銅綠假單胞菌ATCC9027,NCIMB8626,CIP82118ANAEROBICBACTERIUM厭氧細(xì)菌CLOSTRIDIUMSPOROGENES3生孢梭菌ATCC19404,CIP793,NCTC532ORATCC11437FUNGI真菌類CANDIDAALBICANS白色念珠菌ATCC10231,IP4872,NCPF3179ASPERGILLUSNIGER黑曲霉ATCC16404,IP143183,IMI1490071ANALTERNATIVETOSTAPHYLOCOCCUSAUREUSISBACILLUSSUBTILISATCC6633可選擇桿狀菌ATCC6633代替葡萄球狀菌2ANALTERNATIVEMICROORGANISMISMICROCOCCUSLUTEUSKOCURIARHIZOPHILA,ATCC9341可選擇微球菌ATCC9341代替微生物3ANALTERNATIVETOCLOSTRIDIUMSPOROGENES,WHENANONSPOREFORMINGMICROORGANISMISDESIRED,ISBACETROIDESVULGATUSATCC8482當(dāng)需要不產(chǎn)生孢子的微生物時，可選擇普通擬桿菌ATCC8482來代替梭菌。NOTESEEDLOTCULTUREMAINTENANCETECHNIQUESSEEDLOTSYSTEMSAREUSEDSOTHATTHEVIABLEMICROORGANISMSUSEDFORINOCULATIONARENOTMORETHANFIVEPASSAGESREMOVEDFROMTHEORIGINALMASTERSEEDLOT注釋使用種子批培養(yǎng)保護(hù)技術(shù)（種子批體系），以至接種所用的可繁殖微生物不超過五代而遠(yuǎn)離原始主種子批SUITABILITYTESTS適用性試驗(yàn)THEMEDIAUSEDCOMPLYWITHTHEFOLLOWINGTESTS,CARRIEDOUTBEFORE,ORINPARALLEL,WITHTHETESTONTHEPRODUCTTOBEEXAMINED所用培養(yǎng)基和以下試驗(yàn)相符合，它在受檢產(chǎn)品測試之或與之同時進(jìn)行。STERILITY無菌性CONFIRMTHESTERILITYOFEACHSTERILIZEDBATCHOFMEDIUMBYINCUBATINGAPORTIONOFTHEMEDIAATTHESPECIFIEDINCUBATIONTEMPERATUREFOR14DAYSNOGROWTHOFMICROORGANISMSOCCURS確定每批消過毒的培養(yǎng)基的無菌性，在特殊的培養(yǎng)溫度中培育一部份培養(yǎng)基14天，微生物不生長。GROWTHPROMOTIONTESTOFAEROBES,ANAEROBES,ANDFUNGI需氧菌，壓氧菌和真菌類助長試驗(yàn)TESTEACHLOTOFOFREADYPREPAREDMEDIUMANDEACHBATCHOFMEDIUMPREPAREDEITHERFROMDEHYDRATEDMEDIUMORFROMINGREDIENTS1SUITABLESTRAINSOFMICROORGANISMSAREINDICATEDINTABLE1對準(zhǔn)備好的每批培養(yǎng)基，和從干粉培養(yǎng)基或混合粉中準(zhǔn)備的每批培養(yǎng)基進(jìn)行試驗(yàn)表1中顯示出適宜的微生物類。INOCULATEPORTIONSOFFLUIDTHIOGLYCOLLATEMEDIUMWITHASMALLNUMBERNOTMORETHAN100CFUOFTHEFOLLOWINGMICROORGANISMS,USINGASEPARATEPORTIONOFMEDIUMFOREACHOFTHEFOLLOWINGSPECIESOFMICROORGANISMCLOSTRIDIUMSPOROGENES,PSEUDOMONASAERUGINOSA,ANDSTAPHYLOCOCCUSAUREUSINOCULATEPORTIONSOFALTERNATIVEFLUIDTHIOGLYCOLLATEMEDIUMWITHASMALLNUMBERNOTMORETHAN100CFUOFCLOSTRIDIUMSPOROGENESINOCULATEPORTIONSOFSOYBEANCASEINDIGESTMEDIUMWITHASMALLNUMBERNOTMORETHAN100CFUOFTHEFOLLOWINGMICROORGANISMS,USINGASEPARATEPORTIONOFMEDIUMFOREACHOFTHEFOLLOWINGSPECIESOFMICROORGANISMASPERGILLUSNIGER,BACILLUSSUBTILIS,ANDCANDIDAALBICANSINCUBATEFORNOTMORETHAN3DAYSINTHECASEOFBACTERIAANDNOTMORETHAN5DAYSINTHECASEOFFUNGI用一小部份（不超過100CFU）以下微生物，來接種一部份液態(tài)硫乙醇酸鹽培養(yǎng)基，以下每種微生物都分別有一部份培養(yǎng)基梭菌，假單胞菌和葡萄狀球菌。用小數(shù)量的（不超過100CFU）梭菌來嫁接可選擇疏基醋酸液培養(yǎng)基。用小數(shù)量（不超過100CFU）的以下微生物來接種大豆酪蛋白消化物培養(yǎng)基，以下每種微生物使用單獨(dú)的培養(yǎng)基黑曲霉菌，枯草芽孢桿菌和白色念珠菌。在細(xì)菌的情況下培育不超過3天，在真菌類情況下，培育不超過5天。THEMEDIAARESUITABLEIFACLEARLYVISIBLEGROWTHOFTHEMICROORGANISMSOCCURS如果微生物生長明顯出現(xiàn)，培養(yǎng)基適用。STORAGE存儲IFPREPAREDMEDIAARESTOREDINUNSEALEDCONTAINERS,THEYCANBEUSEDFOR1MONTH,PROVIDEDTHATTHEYARETESTEDFORGROWTHPROMOTIONWITHIN2WEEKSOFTHETIMEOFUSEANDTHATCOLORINDICATORREQUIREMENTSAREMETIFSTOREDINTIGHTCONTAINERS,THEMEDIACANBEUSEDFOR1YEAR,PROVIDEDTHATTHEYARETESTEDFORGROWTHPROMOTIONWITHIN3MONTHSOFTHETIMEOFUSEANDTHATTHECOLORINDICATORREQUIREMENTSAREMET如果已準(zhǔn)備培養(yǎng)基存儲于未密封容器中，可使用1個月，使用的兩周時間內(nèi)進(jìn)行助長試驗(yàn)，指示劑的顏色符合要求，可使用1個月。如果存存儲于一個密封的容器中，使用時間的三個月內(nèi)進(jìn)行助長試驗(yàn)，指示劑顏色符合要求，培養(yǎng)基可用1年。DILUTINGANDRINSINGFLUIDSFORMEMBRANEFILTRATION薄膜過濾法的稀釋液和沖洗液FLUIDA液體APREPARATIONDISSOLVE1GOFPEPTICDIGESTOFANIMALTISSUEINWATERTOMAKE1L,FILTERORCENTRIFUGETOCLARIFY,IFNECESSARY,ANDADJUSTTOAPHOF7102DISPENSEINTOCONTAINERS,ANDSTERILIZEUSINGAVALIDATEDPROCESS溶解制劑將1G動物組織胃蛋白酶消化物加入水中制成1L，如果需要，過濾或者離心直到溶液澄清，并將其PH值調(diào)至7102，分配到溶器中，通過驗(yàn)證程序滅菌。PREPARATIONFORPENICILLINSORCEPHALOSPORINSASEPTICALLYADDTOTHEABOVEPREPARATION,IFNECESSARY,AQUANTITYOFSTERILELACTAMASESUFFICIENTTOINACTIVATEANYRESIDUALANTIBIOTICACTIVITYONTHEMEMBRANESAFTERTHESOLUTIONOFTHETESTSPECIMENHASBEENFILTEREDSEEMEDIAFORPENICILLINSORCEPHALOSPORINS青霉素和頭孢子菌的無菌配制將一定量的內(nèi)酰胺酶加入上述所備制劑中，以鈍化在試驗(yàn)樣品溶液過濾后薄膜上任何殘余抗生素活性（見青霉素或頭孢菌素培養(yǎng)基）。FLUIDD液體DTOEACHLOFFLUIDAADD1MLOFPOLYSORBATE80,ADJUSTTOAPHOF7102,DISPENSEINTOCONTAINERS,ANDSTERILIZEUSINGAVALIDATEDPROCESSUSETHISFLUIDFORARTICLESCONTAININGLECITHINOROIL,ORFORDEVICESLABELEDAS“STERILEPATHWAY”每1L液體A就加入1ML聚山梨醇酯80，將PH值調(diào)至7102，并入配到容器中，通過驗(yàn)證程序滅菌。此液體用于含蛋黃素或油脂的藥品，或用于貼著“無菌路徑”的儀器。FLUIDK液體KDISSOLVE50GOFPEPTICDIGESTOFANIMALTISSUE,30GOFBEEFEXTRACT,AND100GOFPOLYSORBATE80INWATERTOMAKE1LADJUSTTHEPHTOOBTAIN,AFTERSTERILIZATION,APHOF6902DISPENSEINTOCONTAINERS,ANDSTERILIZEUSINGAVALIDATEDPROCESS將50G動物組織胃蛋白酶消化液，30G濃縮牛肉汁，和100G聚山梨醇酯80溶解制成1L。調(diào)節(jié)PH值，在滅菌后，達(dá)到6902。分配到容器中，通過驗(yàn)證程序滅菌。VALIDATIONTEST驗(yàn)證測試CARRYOUTATESTASDESCRIBEDBELOWUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDUSINGEXACTLYTHESAMEMETHODS,EXCEPTFORTHEFOLLOWINGMODIFICATIONS按受檢產(chǎn)品無菌試驗(yàn)下同樣的方法，按以下所述進(jìn)行試驗(yàn)操作，除以下更改。MEMBRANEFILTRATION薄膜過濾法AFTERTRANSFERRINGTHECONTENTOFTHECONTAINERORCONTAINERSTOBETESTEDTOTHEMEMBRANE,ADDANINOCULUMOFASMALLNUMBEROFVIABLEMICROORGANISMSNOTMORETHAN100CFUTOTHEFINALPORTIONOFSTERILEDILUENTUSEDTORINSETHEFILTER在將容器內(nèi)溶液或者待測容器中的溶液轉(zhuǎn)移至薄膜后，在用來清洗過濾器最后部份的無菌稀釋液里加入（不超過100CFU）小部份可繁殖微生物的培養(yǎng)基。DIRECTINOCULATION直接接種AFTERTRANSFERRINGTHECONTENTSOFTHECONTAINERORCONTAINERSTOBETESTEDFORCATGUTANDOTHERSURGICALSUTURESFORVETERINARYUSESTRANDSTOTHECULTUREMEDIUM,ADDANINOCULUMOFASMALLNUMBEROFVIABLEMICROORGANISMSNOTMORETHAN100CFUTOTHEMEDIUM在將容器內(nèi)溶液或者待測容器（腸線或其它獸醫(yī)用的外科縫合用線）中的溶液轉(zhuǎn)移至培養(yǎng)基后，在培養(yǎng)基中加入（不超過100CFU）少量的可繁殖微生物。INBOTHCASESUSETHESAMEMICROORGANISMSASTHOSEDESCRIBEDABOVEUNDERGROWTHPROMOTIONTESTOFAEROBES,ANAEROBES,ANDFUNGIPERFORMAGROWTHPROMOTIONTESTASAPOSITIVECONTROLINCUBATEALLTHECONTAINERSCONTAININGMEDIUMFORNOTMORETHAN5DAYS在兩種情況下，按需氧菌，厭氧性生物，和真菌類的助長試驗(yàn)上面所描述，使用同種微生物。執(zhí)行助長試驗(yàn)作為積極控制。對含有培養(yǎng)基的所有容器進(jìn)行不超過五天培育。IFCLEARLYVISIBLEGROWTHOFMICROORGANISMSISOBTAINEDAFTERTHEINCUBATION,VISUALLYCOMPARABLETOTHATINTHECONTROLVESSELWITHOUTPRODUCT,EITHERTHEPRODUCTPOSSESSESNOANTIMICROBIALACTIVITYUNDERTHECONDITIONSOFTHETESTORSUCHACTIVITYHASBEENSATISFACTORILYELIMINATEDTHETESTFORSTERILITYMAYTHENBECARRIEDOUTWITHOUTFURTHERMODIFICATION如果培養(yǎng)后，與沒有產(chǎn)品的控制容器內(nèi)相比，或者與在試驗(yàn)條件下不含抗微生物活性的產(chǎn)品相比，或者與活性已消除產(chǎn)品相比，微生物生長明顯示，那么無菌試驗(yàn)可在不修改下執(zhí)行。IFCLEARLYVISIBLEGROWTHISNOTOBTAINEDINTHEPRESENCEOFTHEPRODUCTTOBETESTED,VISUALLYCOMPARABLETOTHATINTHECONTROLVESSELSWITHOUTPRODUCT,THEPRODUCTPOSSESSESANTIMICROBIALACTIVITYTHATHASNOTBEENSATISFACTORILYELIMINATEDUNDERTHECONDITIONSOFTHETESTMODIFYTHECONDITIONSINORDERTOELIMINATETHEANTIMICROBIALACTIVITY,ANDREPEATTHEVALIDATIONTEST與沒有產(chǎn)品的控制容器內(nèi)相比，或者與在試驗(yàn)條件下含抗微生物活性的產(chǎn)品活性未被消化相比，如果受檢產(chǎn)品的微生物沒有明顯的生長，那么更改條件，以消除抗微生物活性，并重復(fù)驗(yàn)證測試。THISVALIDATIONISPERFORMEDAWHENTHETESTFORSTERILITYHASTOBECARRIEDOUTONANEWPRODUCTANDBWHENEVERTHEREISACHANGEINTHEEXPERIMENTALCONDITIONSOFTHETESTTHEVALIDATIONMAYBEPERFORMEDSIMULTANEOUSLYWITHTHETESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDA當(dāng)新產(chǎn)品進(jìn)行無菌試驗(yàn)時，B當(dāng)測試的試驗(yàn)條件出現(xiàn)變化時,驗(yàn)證執(zhí)行；驗(yàn)證與受檢產(chǎn)品無菌測試可以同時進(jìn)行。TESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINED受檢產(chǎn)品無菌測試NUMBEROFARTICLESTOBETESTED受檢藥品數(shù)UNLESSOTHERWISESPECIFIEDELSEWHEREINTHISCHAPTERORINTHEINDIVIDUALMONOGRAPH,TESTTHENUMBEROFARTICLESSPECIFIEDINTABLE3IFTHECONTENTSOFEACHARTICLEAREOFSUFFICIENTQUANTITYSEETABLE2,THEYMAYBEDIVIDEDSOTHATEQUALAPPROPRIATEPORTIONSAREADDEDTOEACHOFTHESPECIFIEDMEDIANOTEPERFORMSTERILITYTESTINGEMPLOYINGTWOORMOREOFTHESPECIFIEDMEDIAIFEACHARTICLEDOESNOTCONTAINSUFFICIENTQUANTITIESFOREACHMEDIUM,USETWICETHENUMBEROFARTICLESINDICATEDINTABLE3除非此章節(jié)或者專論中的其它地方有特別說明，否則按表3詳細(xì)列出的藥品數(shù)進(jìn)行測試。如果每種藥品的成份足量（見表2），它們將會被分成適當(dāng)?shù)男》莶⒈患拥矫糠葜付ǖ呐囵B(yǎng)基中。注釋進(jìn)行無菌試驗(yàn)時，運(yùn)用兩到三種指定培養(yǎng)基。如果每種培養(yǎng)基中的每種藥品量不足，按表3所示藥品用兩倍。TABLE2MINIMUMQUANTITYTOBEUSEDFOREACHMEDIUM表2每種培養(yǎng)基所使用的最小量QUANTITYPERCONTAINER每個容器的量MINIMUMQUANTITYTOBEUSED除非另有證明及授權(quán)所用的最小量（除非另有證明和授權(quán)）LIQUIDSOTHERTHANANITBIOTICS液體（除抗生素外）LESSTHAN1ML少于1MLTHEWHOLECONTENTSOFEACHCONTAINER每個容器的全部量140MLHALFTHECONTENTSOFEACHCONTAINER,BUTNOTLESSTHAN1ML每個容器容量的一半，但不少于1MLGREATERTHAN40ML,ANDNOTGREATERTHAN100ML大于40ML，但不超過100ML20MLGREATERTHAN100ML超過100ML10OFTHECONTENTSOFTHECONTAINER,BUTQUANTITYPERCONTAINER每個容器的量MINIMUMQUANTITYTOBEUSED除非另有證明及授權(quán)所用的最小量（除非另有證明和授權(quán)）NOTLESSTHAN20ML容器量的10，但不超過20MLANTIBIOTICLIQUIDS液體抗生素1MLOTHERPREPARATIONSSOLUBLEINWATERORINISOPROPYLMYRISTATE其它可溶于水或透汽性油脂的制劑THEWHOLECONTENTSOFEACHCONTAINERTOPROVIDENOTLESSTHAN200MG每個容器的全部容量，不少于200MGINSOLUBLEPREPARATIONS,CREAMS,ANDOINTMENTSTOBESUSPENDEDOREMULSIFIED可溶解制劑，懸浮的或浮化的乳脂和藥膏USETHECONTENTSOFEACHCONTAINERTOPROVIDENOTLESSTHAN200MG每個容器的容量，少于200MGSOLIDS固體LESSTHAN50MG少于50MGTHEWHOLECONTENTSOFEACHCONTAINER每個容器的全部量50MGORMORE,BUTLESSTHAN300MG50MG或更多，但少于300MGHALFTHECONTENTSOFEACHCONTAINER,BUTNOTLESSTHAN50MG每個容器體積的一半，但不少于50MG300MG5G150MGGREATERTHAN5G超過5G500MGDEVICES儀器CATGUTANDOTHERSURGICALSUTURESFORVETERINARYUSE腸線和其它用于獸醫(yī)手術(shù)縫合線3SECTIONSOFASTRANDEACH30CMLONG繩的3段（每斷30CM）SURGICALDRESSING/COTTON/GAUZEINPACKAGES外科手術(shù)敷料/棉花/薄紗（包）100MGPERPACKAGE每包100MGSUTURESANDOTHERINDIVIDUALLYPACKAGEDSINGLEUSEMATERIAL縫合線和其它單獨(dú)包裝使用的材料THEWHOLEDEVICE整個儀器OTHERMEDICALDEVICES其它醫(yī)療儀器THEWHOLEDEVICE,CUTINTOPIECESORDISASSEMBLED整個儀器，切成片或拆分TABLE3MINIMUMNUMBEROFARTICLESTOBETESTEDINRELATIONTOTHENUMBEROFARTICLESINTHEBATCH表3。關(guān)于一批藥品數(shù)中受檢藥品的最小量NUMBEROFITEMSINTHEBATCH一批藥的條款項(xiàng)MINIMUMNUMBEROFITEMSTOBETESTEDFOREACHMEDIUM每種培養(yǎng)基的受檢的項(xiàng)的最小數(shù)除非另有證明及授權(quán)PARENTERALPREPARATIONS非腸道注射用藥的制劑NOTMORETHAN100CONTAINERS不超過100個容器10OR4CONTAINERS,WHICHEVERISTHEGREATER10或4個容器，選用較大者M(jìn)ORETHAN100BUTNOTMORETHAN500CONTAINERS超過100個容器，但不超過500個容器10CONTAINERS10個容器MORETHAN500CONTAINERS超過500個容器2OR20CONTAINERS,WHICHEVERISLESS2或20個容器，選用較小者FORLARGEVOLUMEPARENTERALS大量非腸道注射用藥2OR10CONTAINERS,WHICHEVERISLESS2或20個容器，選用較小者ANTIBIOTICSOLIDS固體抗生素PHARMACYBULKPACKAGES5G散裝藥20CONTAINERS20個容器PHARMACYBULKPACKAGES5G散裝藥6CONTAINERS6個容器BULKSANDBLENDS散裝藥和混合物SEEBULKSOLIDPRODUCTS見固體散裝藥OPHTHALMICANDOTHERNONINJECTABLEPREPARATIONS眼藥和其它非注射制劑NOTMORETHAN200CONTAINERS不超過200個容器5OR2CONTAINERS,WHICHEVERISTHEGREATER5或2個容器，選用較大者M(jìn)ORETHAN200CONTAINERS超過200個容器10CONTAINERS10個容器IFTHEPRODUCTISPRESENTEDINTHEFORMOFSINGLEDOSECONTAINERS,APPLYTHESCHEMESHOWNABOVEFORPREPARATIONSFORPARENTERALUSE如果藥品裝入單劑型容器中，應(yīng)用于以上非腸道注射藥用方案DEVICES儀器CATGUTANDOTHERSURGICALSUTURESFORVETERINARYUSE非腸道注射用藥的制劑2OR5PACKAGES,WHICHEVERISTHEGREATER,UPTOAMAXIMUMTOTALOF20PACKAGES2或5包，選用較大者，至多為20包NOTMORETHAN100ARTICLES不超過200種藥品10OR4ARTICLES,WHICHEVERISGREATER10或4包，選用較大者，MORETHAN100,BUTNOTMORETHAN500ARTICLES超過100，但不超過500種藥品10ARTICLES10包NUMBEROFITEMSINTHEBATCH一批藥的條款項(xiàng)MINIMUMNUMBEROFITEMSTOBETESTEDFOREACHMEDIUM每種培養(yǎng)基的受檢的項(xiàng)的最小數(shù)除非另有證明及授權(quán)MORETHAN500ARTICLES超過500種藥品2OR20ARTICLES,WHICHEVERISLESS2或20包，選用較小者BULKSOLIDPRODUCTS固體散裝藥UPTO4CONTAINERS至多4個容器EACHCONTAINER每個容器MORETHAN4CONTAINERS,BUTNOTMORETHAN50CONTAINERS超過4個容器，但不超過50個容器20OR4CONTAINERS,WHICHEVERISGREATER2或4包，選用較大者M(jìn)ORETHAN50CONTAINERS超過50個容器2OR10CONTAINERS,WHICHEVERISGREATER2或10包，選用較大者IFTHECONTENTSOFONECONTAINERAREENOUGHTOINOCULATETHETWOMEDIA,THISCOLUMNGIVESTHENUMBEROFCONTAINERSNEEDEDFORBOTHTHEMEDIATOGETHER如果一個容器的所裝物足夠接種兩種培養(yǎng)基，此量滿足兩種培養(yǎng)基所需容器數(shù)的量。THETESTMAYBECARRIEDOUTUSINGTHETECHNIQUEOFMEMBRANEFILTRATIONORBYDIRECTINOCULATIONOFTHECULTUREMEDIUMWITHTHEPRODUCTTOBEEXAMINEDAPPROPRIATENEGATIVECONTROLSAREINCLUDEDTHETECHNIQUEOF