灭菌的方法及注意事项ppt课件

1,Sterilization灭菌,AGeneralDiscussionfromCGMPPerspective从CGMP角度的综合讨论,1,2,Outline讨论纲要,PARTITerminology专业用词的定义GMPRequirementsinTheManufactureofSterilePharmaceuticalProducts无菌药品生产过程中GMP的基本要求PARTIIFacilityDesign生产设施的设计HVACSystem空调系统EnvironmentalMonitoring(EM)环境的监视PharmaceuticalWater制药用水Cleaning/Sanitation清洁消毒Personnel无菌区操作人员PARTIIIMethodsofsterilization灭菌方法SterileProductionandvalidation无菌生产和验证TheTrendofSterileProduction无菌产品生产趋势,2,3,PresentationOutline概要,USRegulations美国法规MoistHeatSterilization湿热灭菌DryHeat/Depyrogenation干热/去热原法SterilizationProcessValidation灭菌工艺验证OtherSterilizationMethods其他灭菌方法,3,4,CodeFederalRegulation美国联邦法规,211.84(c)(3)sterileequipment灭菌设备Sterileequipmentandasepticsamplingtechniquesshallbeusedwhennecessary必要时应使用灭菌设备和无菌取样技术。211.94(c)and(d)sterilized(c)Drugproductcontainersandclosuresshallbecleanand,whereindicatedbythenatureofthedrug,sterilizedandprocessedtoremovepyrogenicpropertiestoassurethattheyaresuitablefortheirintendeduse.药品容器和密封系统应清洁并根据药品的性质和要求，进行灭菌，除热原过程以确保预期的用途。(d)Standardsorspecifications,methodsoftesting,and,whereindicated,methodsofcleaning,sterilizing,andprocessingtoremovepyrogenicpropertiesshallbewrittenandfollowedfordrugproductcontainersandclosures.应建立并执行对药品容器和密封系统的规格或质量标准，测试方法，清洁方法，灭菌和除热原过程的相关书面程序。,4,5,CodeFederalRegulation美国联邦法规,211.113(a)and(b)sterilization灭菌(a)Appropriatewrittenprocedures,designedtopreventobjectionablemicroorganismsindrugproductsnotrequiredtobesterile,shallbeestablishedandfollowed.应该建立并执行用于防止非无菌药品被致病菌污染的相关书面程序。(b)Appropriatewrittenprocedures,designedtopreventmicrobiologicalcontaminationofdrugproductspurportingtobesterile,shallbeestablishedandfollowed.Suchproceduresshallincludevalidationofanysterilizationprocess.应该建立并执行用于阻止无菌药品被致病菌污染的相关书面程序。这些程序应包括任何无菌工艺的验证。,5,6,CodeFederalRegulation美国联邦法规,211.167(a)testing（a）测试Foreachbatchofdrugproductpurportingtobesterileand/orpyrogen-free,thereshallbeappropriatelaboratorytestingtodetermineconformancetosuchrequirements.Thetestproceduresshallbeinwritingandshallbefollowed.对无菌和/或无热原的每批药品应进行相应的实验室测试以确定其与符合要求。测试程序应有书面文件并遵照执行。,6,7,SterilizationMethods灭菌方法,MoistHeat:湿热Fordrugsanddevices.Themodeofactionisproteindenaturation.用于药品和设备。作用方式是使蛋白质变性DryHeat:干热Fordepyrogenationandequipmentsterilization.Themodeofactionisproteindenaturation用于去热原和设备灭菌。作用方式是使蛋白质变性EthyleneOxide:氧化乙烯Primarilyfordevices.Themodeofactionisalkylationofnucleicacids.主要用于设备。作用方式是使核酸烷基化Radiation:辐射Primarilyfordevices.ThemodeofactionisDNAstrandbreakage主要用于设备。作用方式是使DNA链破坏Othermethods?其他方法？,7,8,Bioburden生物负荷,Definition:定义Populationofviablemicroorganismsonorinaproductand/orapackage产品和/包装上的活性微生物的数量和类型Mixtureoforganisms有机物混合Indigenousmicroflora地方微植物群Needstobeinactivatedbysterilization需灭菌失活,8,9,Bioburden生物负荷,BioburdenSources生物负荷来源Environment环境Productcontactsurface,personnel,air产品接触表面，人员，空气Materials材料Water,rawmaterials.plastic,paper水，原材料，塑料，纸张CharacteristicsofBioburden生物负荷特点Typesofmicroorganisms微生物类型Resistancetosterilizationprocess对无菌工艺的耐受Numberoforganisms有机物数量,9,10,BiologicalIndicator(BI)生物指示剂,Microbiologicaltestsystemprovidingadefinedresistancetoaspecificsterilizationprocess微生物测试系统对指定灭菌工艺有明确抵抗性。Acharacterizedpreparationofspecificmicroorganismsresistanttoaparticularsterilizationprocess某一确定的微生物(指示剂)应具有对某一特定灭菌工艺的抵抗性,10,11,TypicalBiologicalIndicators典型生物指示剂,Moistheatsterilization湿热灭菌GeobacillusstearothermophilusBacillusstearothermophilusDryheatandEOsterilization干热和环氧乙烷灭菌BacillusatrophaeusBacillussubtilisvar.niger,11,12,FormsofBiologicalIndicators生物指示剂形式,Strips/discsinglassineenvelopes在透明纸信封里的条形板/光盘Strips/discs条/光盘Self-contained独立包装的Ampoules安瓶Stripswithmedium中号条形板Liquidsuspension液体悬浮液Metal金属Fiberglass玻璃纤维,12,13,ChoiceofanAppropriateBI相关生物指示剂的选择,Sterilizationprocess灭菌工艺Cycledesignmethod循环设计方法Productbioburden产品生物负荷Population数量Resistance抵抗性,13,14,BIsPreparedbyUser生物指示剂的准备,Performance性能Resistance抵抗性Population数量Purity纯度D-valueD值Recoverymethods恢复方法Storagerequirements储存要求,14,15,BiologicalIndicatorUse生物指示剂使用,PlaceBIwithin把BI放进Product产品Package包装Sterilizerloadtomonitorprocess灭菌器负荷以监视灭菌工艺Exposetosterilizingconditions暴露在灭菌状态RemoveBIandtest移除BI和测试Countsurvivors生存数量的计算Growth/nogrowthresponse生长/无生长反应,15,16,D-valueD值,TheDvalueisthetime,usuallyinminutes,requiredtoachieveinactivationof90%(oronelogarithm)ofapopulationofthetestmicroorganismatspecifiedconditions.D值是在特定条件下微生物数量降低的90%（或一对数）所用的时间，通常是以分钟为单位的。BacillusstearothermophilushasaDvalue:嗜热脂肪芽胞杆菌的D值2minat121oC20minat110oC0.2-0.3minat130oCOfalltheaspectsofsterilizationvalidation,theDvalueisperhapsthemostimportant.ValidatingaprocesswithoutconsiderationoftheDvalueislargelyineffectiveandisnotacceptablefromCGMPperspective灭菌验证中，D值可能是最重要的。不考虑D值的验证过程多半是无效的且不被CGMP接受。,16,17,Z-valueZ值,Z-value:numberofdegreesoftemperaturerequiredfora1logarithmchangeintheD-valueZ值：D值1对数改变需要的温度数Z=-1/slopeofthethermalresistancecurveZ=-1/热阻力曲线斜率whereSlope=logarithmicchangeinD-value/changeintemperature斜率=D值上对数的改变/温度的改变,17,18,ImpactofZ-valueZ值的影响,Whenz-valueissmall,considerablylessinactivationwillresultbelowreferencetemperatureandconsiderablymoreabovethereferencetemperature当Z值较小时，较低程度的失活将导致温度低于参考温度，较大程度的失活将导致温度高于参考温度。,18,19,TypicalTemperatureProfiles典型温度分布,19,20,TypeofSterilization灭菌类型,MoistHeatSterilization湿热灭菌,20,21,MoistHeatSterilization湿热灭菌,Characteristics:特征Wellunderstoodandwellcharacterizedprocess,firstvalidatedprocessinpharmindustry湿热灭菌是一已被很好地理解并描述灭菌工艺，也是在制药工业中第一个被验证的灭菌工艺Suitableforawidevarietyofapplications适合于较大应用范围Equipmentisreadilyavailable设备很容易从市场购买到Costonaperusebasisislow每次使用基准花费低,21,22,MoistHeatSterilization湿热灭菌,Applications:应用Terminalsterilizationofparentalproduct注射剂的终端灭菌Sterilizationofequipmentandcomponentsforuseinasepticfilling无菌灌装线上设备和配件的灭菌Sterilizationoflaboratorymaterials实验室用材料的灭菌In-situsterilizationofprocesspipingandequipment(SIP)工艺管道和设备在线灭菌,22,23,BasicTypesofMoistHeatSterilization湿热灭菌基本类型,Saturatedsteam饱和蒸汽Autoclaves(self-closing)高压灭菌柜（半封闭）SIP在线灭菌Superheatedwater过热水Spray喷雾Submerged浸没的SIP在线灭菌Steam-air-mixture(SAM)水蒸气空气混合物,23,24,BasicElementsofSterilizationProcessValidation灭菌工艺验证的基本元素,Emptyvesselheatdistribution空容器热分布Heatdistributionandpenetration热分布和渗透3.Microbiologicalchallenges微生物挑战,24,25,SteamSterilizationValidation:Prerequisites蒸汽灭菌验证：前提,OQforanautoclave:高压灭菌柜运行确认Emptychambertemperaturemappingwithin1.0oCofthemean空腔体温度分布图在平均值的1.0oC内Chamberintegritytest(noleaking)腔体完整性测试CertificationofHEPAfiltrationontheairusedtobreakvacuumorintegritytestingoftheventfilter用于隔断真空或通气过滤器完整性测试的空气HEPA过滤认证RequirementsforSIP在线灭菌要求Temperaturemapping温度分布图Anintegritytest,whereappropriate相关完整性测试UseofBI生物指示剂的使用Allcriticalinstrumentsmustbecalibrated所有关键仪器需校验,25,26,SteamSterilizationValidation:Prerequisites蒸汽灭菌验证：前提,Acceptabletestresultsfornon-condensablegases,super-heatedsteamanddrynessshouldbeobtainedforthecleansteamusedfortheautoclave/SIP应获得用于高压灭菌柜/SIP的洁净蒸汽中的不凝气体，过热蒸汽及干燥度的可接受测试结果5.ToolsfortheconductofthePQstudy:进行PQ研究的工具BIwith106sporesandknownDandZvaluesBI有106个孢子，已知D值和Z值Temperaturesensors温度传感器Recordingdevicecapableofsupporting12tempsensorswithanaccuracyof0.5oC,recordingdataeveryminuteorless记录设备能支撑12温度传感器，精度0.5oC，每分钟或间隔更短时间记录数据Meansofintroducingtempsensorsintotheautoclave/SIP将温度传感器导入高压灭菌柜/SIP的方法,26,27,SteamSterilizationValidation:PreparationofPQprotocol蒸汽灭菌验证：PQ方案的准备,Aprotocolshallbepreparedfor:对于下列各项应建立方案：Newautoclave/SIP新高压灭菌柜/SIPNewloadingpatternsorproductconfigurations新装料方式或产品配置Changestoexistingpatterns对现有装料方式的变更Changestooperationcycleparameters对运行周期参数的变更Majorchangetoequipmentasdirectedbychangecontrol变更控制要求的设备主要变更,27,28,SteamSterilizationValidation:PreparationofPQprotocol蒸汽灭菌验证：PQ方案的准备,Theprotocolmayinclude:方案可能包括：Objectivesofthevalidationstudy验证研究的目的Identificationanddescriptionofthesterilizeranditsprocesscontrols灭菌器的识别和说明及工艺控制IdentificationofSOPsfortheprocessequipment工艺设备SOP的识别DescriptionoforSOPreferenceforinstrumentcalibrationprocedures仪器校验程序的说明或SOP参考Identificationofcalibrationproceduresfortemp-monitoringequipment,whichincludeatwopointpre-runcalibrationandapost-runverificationforeachrun温度监测设备校验程序的识别，包括一个两点预运行校验和每次运行后的确认Processparameteracceptancecriteria工艺参数的验收标准,28,29,SteamSterilizationValidation:PreparationofPQprotocol蒸汽灭菌验证：PQ方案的准备,Adescriptionofthefollowing:以下说明Biodurdendeterminationstudies生物负荷确认研究Emptychamberheatdistributionstudies(1oC)空腔体热分布研究（1oC）Loadedchamber(includingLoadconfiguration,maxladingandminloading)heatpenetrationstudies满载腔体（包括装载配置,最大和最少）热穿透研究Containermappingstudies(maynotneededif100mL)容器分布图研究（如果容量3logreduction大于3个对数减少值,43,44,AdvantagesofDry-heatDepyrogenation干热去热原的优点,Inactivatespyrogenswhilesterilization灭菌时使热原失活Materialsdryattheendofcycle循环结束时物料干燥Corrosiveeffectsareminimal腐蚀性最小Conveyorsystemsallowforhighertempsandshorterdwelltime传送系统允许更高温度和更短的停留时间,44,45,DisadvantagesofDry-heatDepyrogenation干热去热原的缺点,Slowprocess(airisapoorconductor)降低工艺速度（空气是不良导体）Heatpenetrationslowerthansteam热穿透比蒸汽慢Ratevaries(slowforglass,rapidforstainlesssteel)速度各异（玻璃慢，不锈钢快）Heatmustpenetratetoinnersurfaceviaconduction热必须通过传导穿透到内表面Layeringcanoccurduetodifferencesinairdensitywithtemperature;mechanicalcirculationneeded空气密度，温度不同可能导致分层，需要机械循环Heatdegradationlimitsmaterials热降解限制材料Contractionduringcoolingmaydrawcontaminants冷却过程中接触可能引起污染,45,46,DepyrogenationbyTunnels通过烘箱去热原,HEPA-filtered,verticallaminarairflowinheatingandcoolingzones,orradiantheatersinheatingzoneandverticallaminarairflowincoolingzone在加热和冷却区经HEPA过滤的垂直层流气流，或加热区的辐射加热器和冷却区的垂直层流气流Conveyorbelttoprovidein-linecontinuousflowofsterileglasswaretoasepticarea传送带提供连续在线的无菌玻璃器皿到无菌区的流动Limitedtoonetypeofloadatatime一次限用一种装载方式Problemtocontrolspeedmatchotherlineequipment(filler)控制速度以匹配其他线设备（灌装机）的问题Difficulttoachieveuniformheating,andheatsourcemaygenerateparticles很难达到均匀加热，热原可能产生颗粒Largeproductvolumeneededtojustifytunnel调整烘箱要求大的产品容量,46,47,ValidationofDepyrogenationCycles去热原循环验证,Runengineeringtrialstofindcoldspots进行试运行以找到冷点Runtrialstodetermineworsecasevial,e.g.,thickestglass,packingeffects试车以确定最差状况的玻璃瓶，如最厚的玻璃，紧束效应Procureorprepareendotoxinindicators获得或制备内毒素指示剂Run3fullloadswithendotoxinindicatorsinplaceatcoldspotsandrandomsites进行3次满载运行，内毒素指示剂置于冷点和随机位置Acceptancecriteria验收标准min.3logreductioninEUforspikedvials带孔玻璃瓶3个对数减少值，单位为EU3successfulrunswithcontrolsanddocumentation受控并记录的3次成功运行,47,CaseStudy实例分析DryHeatDepyrogenationTunnel干热去热原隧道烘箱,ValidationAcceptanceCriteria验证验收标准Aminimumofthreesuccessfulvalidationrunsmustbeperformed.必须至少进行三次成功的验证Distributionthermocoupletemperaturesmustbewithin15oCofthesetpointtemperature,afterstabilization.稳定后，分配热电偶温度必须在设定温度的15oC内AminimumcumulativeFHvalueoftwelveminutesmustbedemonstratedforeachpenetrationthermocoupleattheendofeachcycle.每次循环结束时，必须证明每个渗透热电偶12分钟内的最少累计FH值Aminimum3-logreductionofendotoxinmustbedemonstratedforeachendotoxinspikedvialexposedtothedepyrogenationcycle.必须证明暴露于去热原循环的每个内毒素长颈瓶至少有3个内毒素对数减少值,48,CaseStudy实例分析DryHeatDepyrogenationTunnel干热去热原隧道烘箱,ValidationAcceptanceCriteria验证验收标准ThecontrollingRTD(hotzoneEntranceRTD)andthethermocoupleadjacenttothecontrollingRTDmustbewithin10oCofeachotherduringtheexposure/dwellphaseafterstabilization.稳定后，暴露/停留阶段控制RTD（高温区入口RTD）和靠近控制RTD的热电偶之间温差必须在10CAminimumof20of26thermocouplesmustbefunctionalandmeetpostcyclecalibrationcheckcriteriaperXXXXXSOP.26个热电偶中至少有20个能起作用，满足XXXXXSOP后循环校验检查标准。,49,50,OtherSterilizationMethods其他灭菌方法,EthyleneOxideSterilization氧化乙烯灭菌Agaseouschemicalagent一种气态化学剂Mo