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/或日本藥典的相關內容一致。不一致的章節(jié)用（）符號來表示，以標明其事實。THEFOLLOWINGPROCEDURESAREAPPLICABLEFORDETERMININGWHETHERAPHARMACOPEIALARTICLEPURPORTINGTOBESTERILECOMPLIESWITHTHEREQUIREMENTSSETFORTHINTHEINDIVIDUALMONOGRAPHWITHRESPECTTOTHETESTFORSTERILITYPHARMACOPEIALARTICLESARETOBETESTEDBYTHEMEMBRANEFILTRATIONMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDWHERETHENATUREOFTHEPRODUCTPERMITSIFTHEMEMBRANEFILTRATIONTECHNIQUEISUNSUITABLE,USETHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDALLDEVICES,WITHTHEEXCEPTIONOFDEVICESWITHPATHWAYSLABELEDSTERILE,ARETESTEDUSINGTHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODPROVISIONSFORRETESTINGAREINCLUDEDUNDEROBSERVATIONANDINTERPRETATIONOFRESULTS以下程序用來確認藥典藥品是否按單獨記錄所列相關無菌測試要求執(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。除設備路徑標明無菌外，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果觀察資料及說明。BECAUSESTERILITYTESTINGISAVERYEXACTINGPROCEDURE,WHEREASEPSISOFTHEPROCEDUREMUSTBEENSUREDFORACORRECTINTERPRETATIONOFRESULTS,ITISIMPORTANTTHATPERSONNELBEPROPERLYTRAINEDANDQUALIFIEDTHETESTFORSTERILITYISCARRIEDOUTUNDERASEPTICCONDITIONSINORDERTOACHIEVESUCHCONDITIONS,THETESTENVIRONMENTHASTOBEADAPTEDTOTHEWAYINWHICHTHESTERILITYTESTISPERFORMEDTHEPRECAUTIONSTAKENTOAVOIDCONTAMINATIONARESUCHTHATTHEYDONOTAFFECTANYMICROORGANISMSTHATARETOBEREVEALEDINTHETESTTHEWORKINGCONDITIONSINWHICHTHETESTSAREPERFORMEDAREMONITOREDREGULARLYBYAPPROPRIATESAMPLINGOFTHEWORKINGAREAANDBYCARRYINGOUTAPPROPRIATECONTROLS因為無菌測試是一個極其嚴格的測試，必需確保滅菌程序以求結果的正確詮釋，所以對人員進行相應培訓以至合格就顯得極為重要。無菌測試在無菌條件下進行。為了達到這樣的環(huán)境，應改變周圍的測試環(huán)境至無菌測試環(huán)境。應采取預防措施避免污染，并且此措施不影響測試中有機體的暴露。無菌測試的工作環(huán)境應靠在工作區(qū)取樣來接受定期監(jiān)控，同時進行適當控制。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，或者是已經過滅菌的。它主要是由滅菌過程和無菌加工程序認證來確定的。WHENEVIDENCEOFMICROBIALCONTAMINATIONINTHEARTICLEISOBTAINEDBYTHEAPPROPRIATEPHARMACOPEIALMETHOD,THERESULTSOOBTAINEDISCONCLUSIVEEVIDENCEOFFAILUREOFTHEARTICLETOMEETTHEREQUIREMENTSOFTHETESTFORSTERILITY,EVENIFADIFFERENTRESULTISOBTAINEDBYANALTERNATIVEPROCEDUREFORADDITIONALINFORMATIONONSTERILITYTESTING,SEESTERILIZATIONANDSTERILITYASSURANCEOFCOMPENDIALARTICLES1211當通過相應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果，以此得到的結果為藥品不符合無菌測試要求的最終結果。無菌測試的詳細信息見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備培養(yǎng)基，或無水配方，當按生產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。按驗證方法對培養(yǎng)基進行滅菌。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值會達到7102。如果有必要過濾的話，加熱溶液但不用達到沸點，熱氣通過潮濕的過濾紙時進行過濾。加入刃天青鈉溶液，并攪拌混合，將培養(yǎng)基放入適當?shù)娜萜髦校巳萜骺梢燥@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。用驗證方法進行滅菌。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，并防止有菌氣體產生并進入容器，液態(tài)硫乙醇酸鹽培養(yǎng)基就在32525下進行培育。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，使用前允許進行冷卻。滅菌后PH值為7102。在厭氧條件下進行培育，以求培育期的持續(xù)。ALTERNATIVEFLUIDTHIOGLYCOLLATEMEDIUMISTOBEINCUBATEDAT32525可選擇疏基醋酸液培養(yǎng)基應在32525下進行培育。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的氫氧化鈉來調節(jié)PH值，以至在滅菌后其PH值達到7302。過濾，如果有必要進行澄清時，將漏斗放入相應的容器中，并用驗證方法來進行滅菌。在一個無菌密封容器中2和25下進行存儲，直至使用。SOYBEANCASEINDIGESTMEDIUMISTOBEINCUBATEDAT22525大豆酪蛋白消化物培養(yǎng)基在22525下進行培育。MEDIAFORPENICILLINSORCEPHALOSPORINS青霉素或頭孢菌素培養(yǎng)基WHERESTERILITYTESTMEDIAARETOBEUSEDINTHEDIRECTINOCULATIONOFTHECULTUREMEDIUMMETHODUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINED,MODIFYTHEPREPARATIONOFFLUIDTHIOGLYCOLLATEMEDIUMANDTHESOYBEANCASEINDIGESTMEDIUMASFOLLOWSTOTHECONTAINERSOFEACHMEDIUM,TRANSFERASEPTICALLYAQUANTITYOFLACTAMASESUFFICIENTTOINACTIVATETHEAMOUNTOFANTIBIOTICINTHESPECIMENUNDERTESTDETERMINETHEQUANTITYOFLACTAMASEREQUIREDTOINACTIVATETHEANTIBIOTICBYUSINGALACTAMASEPREPARATIONTHATHASBEENASSAYEDPREVIOUSLYFORITSPENICILLINORCEPHALOSPORININACTIVATINGPOWERNOTESUPPLEMENTEDLACTAMASEMEDIACANALSOBEUSEDINTHEMEMBRANEFILTRATIONTEST當用受檢藥品無菌試驗測試的培養(yǎng)基直接接種方法來進行無菌試驗時，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，將足夠的內酰胺酶轉移到每種培養(yǎng)基的容器中，來鈍化試驗樣品中的抗生素。用以前化驗青霉素或頭孢菌素鈍化力的內酰胺酶制劑來確定需要鈍化抗生素的內酰胺酶的量。ALTERNATIVELYINANAREACOMPLETELYSEPARATEFROMTHATUSEDFORSTERILITYTESTING,CONFIRMTHATANAPPROPRIATEAMOUNTOFLACTAMASEISINCORPORATEDINTOTHEMEDIUM,FOLLOWINGEITHERMETHODUNDERVALIDATIONTEST,USINGLESSTHAN100COLONYFORMINGUNITSCFUOFSTAPHYLOCOCCUSAUREUSSEETABLE1ASTHECHALLENGETYPICALMICROBIALGROWTHOFTHEINOCULATEDCULTUREMUSTBEOBSERVEDASACONFIRMATIONTHATTHELACTAMASECONCENTRATIONISAPPROPRIATE另外（在另一個完全與無菌試驗無關的領域），確定適當量內酰胺酶混合于培養(yǎng)基中，接著用驗證測試中的方法，用不少于100CFU葡萄狀球菌（見表1）做為挑戰(zhàn)性實驗接種的培養(yǎng)菌的典型微生物生長應接受觀察，以確定內酰胺酶的濃渡適當。TABLE1STRAINSOFTHETESTMICROORGANISMSSUITABLEFORUSEINTHEGROWTHPROMOTIONTESTANDTHEVALIDATIONTEST表1。促生長試驗和驗證試驗中的可用到微生物菌株AEROBICBACTERIA需氧細菌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當需要不產生孢子的微生物時，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)保護技術（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離原始主種子批SUITABILITYTESTS適用性試驗THEMEDIAUSEDCOMPLYWITHTHEFOLLOWINGTESTS,CARRIEDOUTBEFORE,ORINPARALLEL,WITHTHETESTONTHEPRODUCTTOBEEXAMINED所用培養(yǎng)基和以下試驗相符合，它在受檢產品測試之或與之同時進行。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TESTEACHLOTOFOFREADYPREPAREDMEDIUMANDEACHBATCHOFMEDIUMPREPAREDEITHERFROMDEHYDRATEDMEDIUMORFROMINGREDIENTS1SUITABLESTRAINSOFMICROORGANISMSAREINDICATEDINTABLE1對準備好的每批培養(yǎng)基，和從干粉培養(yǎng)基或混合粉中準備的每批培養(yǎng)基進行試驗表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的培養(yǎng)基黑曲霉菌，枯草芽孢桿菌和白色念珠菌。在細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，使用的兩周時間內進行助長試驗，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進行助長試驗，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值調至7102，分配到溶器中，通過驗證程序滅菌。PREPARATIONFORPENICILLINSORCEPHALOSPORINSASEPTICALLYADDTOTHEABOVEPREPARATION,IFNECESSARY,AQUANTITYOFSTERILELACTAMASESUFFICIENTTOINACTIVATEANYRESIDUALANTIBIOTICACTIVITYONTHEMEMBRANESAFTERTHESOLUTIONOFTHETESTSPECIMENHASBEENFILTEREDSEEMEDIAFORPENICILLINSORCEPHALOSPORINS青霉素和頭孢子菌的無菌配制將一定量的內酰胺酶加入上述所備制劑中，以鈍化在試驗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值調至7102，并入配到容器中，通過驗證程序滅菌。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。調節(jié)PH值，在滅菌后，達到6902。分配到容器中，通過驗證程序滅菌。VALIDATIONTEST驗證測試CARRYOUTATESTASDESCRIBEDBELOWUNDERTESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDUSINGEXACTLYTHESAMEMETHODS,EXCEPTFORTHEFOLLOWINGMODIFICATIONS按受檢產品無菌試驗下同樣的方法，按以下所述進行試驗操作，除以下更改。MEMBRANEFILTRATION薄膜過濾法AFTERTRANSFERRINGTHECONTENTOFTHECONTAINERORCONTAINERSTOBETESTEDTOTHEMEMBRANE,ADDANINOCULUMOFASMALLNUMBEROFVIABLEMICROORGANISMSNOTMORETHAN100CFUTOTHEFINALPORTIONOFSTERILEDILUENTUSEDTORINSETHEFILTER在將容器內溶液或者待測容器中的溶液轉移至薄膜后，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上面所描述，使用同種微生物。執(zhí)行助長試驗作為積極控制。對含有培養(yǎng)基的所有容器進行不超過五天培育。IFCLEARLYVISIBLEGROWTHOFMICROORGANISMSISOBTAINEDAFTERTHEINCUBATION,VISUALLYCOMPARABLETOTHATINTHECONTROLVESSELWITHOUTPRODUCT,EITHERTHEPRODUCTPOSSESSESNOANTIMICROBIALACTIVITYUNDERTHECONDITIONSOFTHETESTORSUCHACTIVITYHASBEENSATISFACTORILYELIMINATEDTHETESTFORSTERILITYMAYTHENBECARRIEDOUTWITHOUTFURTHERMODIFICATION如果培養(yǎng)后，與沒有產品的控制容器內相比，或者與在試驗條件下不含抗微生物活性的產品相比，或者與活性已消除產品相比，微生物生長明顯示，那么無菌試驗可在不修改下執(zhí)行。IFCLEARLYVISIBLEGROWTHISNOTOBTAINEDINTHEPRESENCEOFTHEPRODUCTTOBETESTED,VISUALLYCOMPARABLETOTHATINTHECONTROLVESSELSWITHOUTPRODUCT,THEPRODUCTPOSSESSESANTIMICROBIALACTIVITYTHATHASNOTBEENSATISFACTORILYELIMINATEDUNDERTHECONDITIONSOFTHETESTMODIFYTHECONDITIONSINORDERTOELIMINATETHEANTIMICROBIALACTIVITY,ANDREPEATTHEVALIDATIONTEST與沒有產品的控制容器內相比，或者與在試驗條件下含抗微生物活性的產品活性未被消化相比，如果受檢產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，并重復驗證測試。THISVALIDATIONISPERFORMEDAWHENTHETESTFORSTERILITYHASTOBECARRIEDOUTONANEWPRODUCTANDBWHENEVERTHEREISACHANGEINTHEEXPERIMENTALCONDITIONSOFTHETESTTHEVALIDATIONMAYBEPERFORMEDSIMULTANEOUSLYWITHTHETESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINEDA當新產品進行無菌試驗時，B當測試的試驗條件出現(xiàn)變化時,驗證執(zhí)行；驗證與受檢產品無菌測試可以同時進行。TESTFORSTERILITYOFTHEPRODUCTTOBEEXAMINED受檢產品無菌測試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詳細列出的藥品數(shù)進行測試。如果每種藥品的成份足量（見表2），它們將會被分成適當?shù)男》莶⒈患拥矫糠葜付ǖ呐囵B(yǎng)基中。注釋進行無菌試驗時，運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所用的最小量（除非另有證明和授權）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所用的最小量（除非另有證明和授權）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)手術縫合線3SECTIONSOFASTRANDEACH30CMLONG繩的3段（每斷30CM）SURGICALDRESSING/COTTON/GAUZEINPACKAGES外科手術敷料/棉花/薄紗（包）100MGPERPACKAGE每包100MGSUTURESANDOTHERINDIVIDUALLYPACKAGEDSINGLEUSEMATERIAL縫合線和其它單獨包裝使用的材料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。關于一批藥品數(shù)中受檢藥品的最小量NUMBEROFITEMSINTHEBATCH一批藥的條款項MINIMUMNUMBEROFITEMSTOBETESTEDFOREACHMEDIUM每種培養(yǎng)基的受檢的項的最小數(shù)除非另有證明及授權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MORETHAN100BUTNOTMORETHAN500CONTAINERS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MORETHAN200CONTAINERS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MINIMUMNUMBEROFITEMSTOBETESTEDFOREACHMEDIUM每種培養(yǎng)基的受檢的項的最小數(shù)除非另有證明及授權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MORETHAN50CONTAINERS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