灭菌的方法及注意事项-课件

11Sterilization

灭菌AGeneralDiscussionfromCGMPPerspective从CGMP角度的综合讨论11Sterilization

灭菌122Outline

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22Outline讨论纲要PARTI23PresentationOutline

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3PresentationOutline

概要USReg34CodeFederalRegulation

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。4CodeFederalRegulation

美国联邦法45CodeFederalRegulation

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。5CodeFederalRegulation

美国联邦法56CodeFederalRegulation

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。6CodeFederalRegulation

美国联邦法67SterilizationMethods

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？7SterilizationMethods

灭菌方法Moi78Bioburden

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8Bioburden

生物负荷Definition:定义89Bioburden

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9Bioburden

生物负荷BioburdenSourc910BiologicalIndicator(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10BiologicalIndicator(BI)

生物1011TypicalBiologicalIndicators

典型生物指示剂Moistheatsterilization

湿热灭菌GeobacillusstearothermophilusBacillusstearothermophilusDryheatandEOsterilization干热和环氧乙烷灭菌BacillusatrophaeusBacillussubtilisvar.niger11TypicalBiologicalIndicator1112FormsofBiologicalIndicators

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12FormsofBiologicalIndicato1213ChoiceofanAppropriateBI

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13ChoiceofanAppropriateBI

1314BIsPreparedbyUser

生物指示剂的准备Performance

性能Resistance抵抗性Population数量Purity纯度D-

value

D值Recoverymethods恢复方法Storagerequirements储存要求14BIsPreparedbyUser

生物指示剂的准1415BiologicalIndicatorUse

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15BiologicalIndicatorUse

生物指1516D-value

D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2 min at121oC20 min at110oC0.2-0.3min at130oC

Ofalltheaspectsofsterilizationvalidation,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。16D-value

D值TheDvalueisth1617Z-value

Z值Z-value:numberofdegreesoftemperaturerequiredfora1logarithmchangeintheD-value

Z值：D值1对数改变需要的温度数Z=-1/slopeofthethermalresistancecurve

Z=-1/热阻力曲线斜率

where Slope=logarithmicchangeinD-value/changeintemperature

斜率=

D值上对数的改变/温度的改变17Z-value

Z值Z-value:numbero1718ImpactofZ-value

Z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。18ImpactofZ-value

Z值的影响When1819TypicalTemperatureProfiles

典型温度分布19TypicalTemperatureProfiles1920TypeofSterilization

灭菌类型MoistHeatSterilization湿热灭菌20TypeofSterilization

灭菌类型Mo2021MoistHeatSterilization

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21MoistHeatSterilization

湿热灭2122MoistHeatSterilization

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

22MoistHeatSterilization

湿热灭2223BasicTypesofMoistHeatSterilization

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23BasicTypesofMoistHeatS2324BasicElementsofSterilizationProcessValidation

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24BasicElementsofSterilizat2425SteamSterilizationValidation:Prerequisites

蒸汽灭菌验证：前提OQforanautoclave:高压灭菌柜运行确认Emptychambertemperaturemappingwithin±1.0oCofthemean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25SteamSterilizationValidati2526SteamSterilizationValidation: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BIwith106sporesandknownDandZvalues

BI有106个孢子，已知D值和Z值Temperaturesensors温度传感器Recordingdevicecapableofsupporting≥12tempsensorswithanaccuracyof≥0.5oC,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26SteamSterilizationValidati2627SteamSterilizationValidation: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27SteamSterilizationValidati2728SteamSterilizationValidation: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28SteamSterilizationValidati2829SteamSterilizationValidation: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(maynotneededif<100mL)

容器分布图研究（如果容量<100mL，可能不需要）Microbiologicalchallengestudies微生物挑战研究Evaluationofdrugproductcoolingwater(whereapplicable)

药品冷却水评估（若适用）Evaluationofventfiltermembranesassociatedwiththesterilizer与灭菌器相关的通气过滤膜的评估29SteamSterilizationValidati2930SteamSterilizationValidation:PreparationofPQprotocol

蒸汽灭菌验证：PQ方案的准备4. Descriptionofthetempsensorplacementwithintheload在负荷范围内温度传感器位置的说明Onenexttothetempcontrollingsensorforautoclave.一1个接近高压灭菌柜的温度控制传感器NLT12sensorsforautoclave

不少于12个传感器用于高压灭菌柜NLT3sensorsforSIP不少于3传感器用于在线灭菌Ifthetempcontrollingsensorisnotinthedrain,anadditionalsensorshallbeplacedinthedrain

如果温度控制传感器不在排水口中，应置另一个传感器于排水口中Aminimum#ofsensorsmeetingthecalibrationandverificationshallbeestablished.Anysensorthatfailattheendofthestudyshallbeinvestigated.Anysensorlocatednexttothetempcontrollingsensororinthedrainshallmeetthepreandpostcalibrationaspartoftheacceptancecriteriaforasuccessfulrun应建立符合校验和验证的最少#传感器。最终任何传感器的失败应调查研究。任何临近温度控制传感器或在排水口的传感器应符合预校验和后校验，这是成功运行的验收标准的一部分。30SteamSterilizationValidati3031SteamSterilizationValidation:PreparationofPQprotocol

蒸汽灭菌验证：PQ方案的准备5. Acceptancecriteriafortemp.pressure,Fowhereappropriate,vacuumanddestructionsofBIs

温度压力，相关Fo，真空及生物指示剂灭活的验收标准6. Arequirementfor3consecutive,successfulruns

3次连续成功运行的要求

Thedrainisthecoldestpointandisoutsideofthesterilizationzone,notincludedinthecalculationofmeanchambertemperatureandnotsubjecttochamberdistributionrequirement.

排水口是最冷点，在灭菌区外，且不包括在平均腔体温度计算内，不受限于腔体分布要求

Thermocouplesshouldbeplacedatthesamelocationforemptychamberandloadedchamberheatdistributionstudies.

热电偶应置于相同位置，进行空腔体和满载腔体热分布研究

Uniformityisexpectedonlyatthesteadystate.

仅在稳定状态下要求均一性31SteamSterilizationValidati3132SteamSterilizationValidation:ExecutionofPQ

蒸汽灭菌验证：PQ执行1. Temperaturesensorshall:温度传感器应：Forheatpenetrationdata,belocatedsuchthattheypenetratetheequipment,ortheproductcontainerandareimmersedintheproduct对于热穿透数据，应置于能穿透设备或产品容器处，并包含在产品中Forheatdistributiondata,beevenlydistributedwithintheloadorsystemandshallnotcontactmetalsurface

对于热分布数据，平均分布在负荷范围内或系统中，且不能接触金属面2. TheBIshall:生物指示剂应：Beplacednearthetipofthetemperaturesensorusedforpenetration置于靠近用于穿透的温度传感器的顶端IncludetheidentificationthatlinkstothenumberoftheappropriatetemperaturesensoroftheBI

包括与相关BI温度传感器数量有关的识别TheDvalueoftheBIshallbedeterminedintheproduct

BI的D值应在生产中确定32SteamSterilizationValidati3233SteamSterilizationValidation:

ExecutionofPQ

蒸汽灭菌验证：PQ执行TheautoclaveshallbeloadedandoperatedinaccordancewithSOP高压灭菌器应按照SOP进行装载并运行Using121.1oCasthebasetemperatureand10oCastheZ-value,Foshallbecalculated121.1oC作为基底温度，10oC作为Z值，Fo应为计算值AcceptancecriteriashallbebasedonnogrowthonexposedBIsandgrowthonthepositivecontrol

BI验收标准应依据暴露的BI上未生长和阳性控制BI上的生长Failuretomeetacceptancecriteriashallrequirecycleparametermodifications,loadpatternmodificationand/orequipmentrepairs/corrections

未达到验收标准应要求周期参数变更，负荷曲线图变更和/或设备修理/修正33SteamSterilizationValidati3334CaseStudy实例分析

AutoclaveSterilizationValidation

高压蒸汽灭菌验证Table1 AcceptanceCriteriaofEmptyChamberHeatDistribution表1空腔体热分布验收标准No.AcceptanceCriteria验收标准1The12TCsdistributedthroughoutthechambermustbewithin±1ºCofthemeanthermocoupletemperatureeachminuteafterstabilizationthroughoutthedurationofthesterilizationphase.整个灭菌阶段，稳定后，每分钟分布在腔体内的12个TC必须在平均热电偶温度的±1ºC

2Dwelltimenotlessthanthepre-setsterilizationtimeof40minutes.停留时间不少于预设定的灭菌时间40分钟3TheTCsmonitoringthedrainandchamberRTDsmaynotfailpostcheckcalibration.TenofthetwelveTCsmustbefunctionalattheendofthecycle.监测排水和腔体RTD的TC不应在实验后校验失败。循环结束时，12个TC中的10个必须在实验后校验时功能完好34CaseStudy实例分析

AutoclaveSt3435CaseStudy

实例分析

AutoclaveSterilizationValidation

高压蒸汽灭菌验证Table2

AcceptanceCriteriaforHeatDistribution表2热分布验收标准No.AcceptanceCriteria验收标准1Dwelltimemustbenotlessthanthepre-setsterilizationtimeof15minutes停留时间不得少于预设定的灭菌时间15分钟2ThedrainPT-100andTCinthedrainmustbewithin1.0°Cofeachotherduringthestabilizedsterilizationperiod.在稳定灭菌期间，排水管PT-100和排水管中TC之间温差必须在1.0oC之内3Thetemperaturedeviationduringholdingtimesdoesnotexceed-1°Cand+2°Cfromthesetpoint(duringthestabilizationperiod)停留时间的温度偏差不超过-1oC及设定点的+2oC（在稳定阶段）4Atleast10TCsof12thermocouplesmustbefunctionalattheendoftherun循环结束时，12个TC中的10个必须功能完好35CaseStudy实例分析

AutoclaveSt3536CaseStudy实例分析

AutoclaveSterilizationValidation

高压蒸汽灭菌验证Table3

AcceptanceCriteriaforHeatPenetration表3热分布验收标准No.AcceptanceCriteria验收标准1F0mustbenotlessthanpresettimeof15minutesF0不得低于预设定的15分钟2ThedrainPT-100andTCinthedrainmustbewithin1.0°Cofeachotherduringthestabilizedsterilizationperiod.在稳定灭菌期间，排水管PT-100和排水管中TC之间温差必须在1.0°C之内3Thetemperaturesensorsremainedinpositionattheendofcycle循环结束时温度传感器仍然在位4Theitemscontainingsensorsareintact;bottleshavenotleaked,burstorbroken装传感器的器具完整；瓶子不漏，爆裂或破碎5Nomorethan1%ofthebottleshasburstorbroken爆裂或破碎的瓶不超过1%6Thetemperaturemeasuredinanyfluidcontainersisnotgreaterthan80ºCattheendofthecycle循环结束时，任何液体容器中测量的温度不超过80ºC7Thetemperaturedeviationduringholdingtimedoesnotexceed-1°Cand+2°Cfromthesetpointduringthestabilizedsterilizationperiod停留时间的温度偏差不超过-1°C及稳定灭菌阶段设定点的+2°C8Nosporesurvivalofthebiologicalindicators生物指示器上无孢子生存9Atleast10TCsof12thermocouplesarefunctionalattheendoftherunforthemaximum&minimumloadconfigurations,respectively每次最大&最小装载配置结束时，12个热电偶中至少10个TC起作用10Autoclaveandvalidationtimersmustbesynchronized高压灭菌柜和确认计时器必须同步36CaseStudy实例分析

AutoclaveSt3637DryHeatSterilization干热灭菌

Depyrogenation去热原37DryHeatSterilization干热灭菌3738TerminologyandDefinitions

术语和定义Pyrogen:Feverproducingsubstance:endotoxin,viruses,fungi,toxinfromgram(+)andgram(-)bacterials,peptiglycan

热原：热产生的物质：内毒素，病毒，真菌，革兰氏阳性和阴性细菌产生的毒素，肽糖Endotoxin:Componentfromoutermembraneofgram(-)bacterial,mostpotentpyrogens,LPSinstructure

内毒素：革兰氏阴性细菌外膜的成分，大多数有效热原，结构中的LPSEnvironmental:complexofprotein,carbohydrate&lipid.

环境：复合蛋白质，碳水化合物&脂类

Purified:lipopolysacchride,usedinendotoxinstandards

提纯后：脂多糖，用于内毒素标准38TerminologyandDefinitions3839SourcesofPyrogeninParenterals

注射用药热原的来源Watersystemsandresincolumns

水系统和树脂交换柱APIsproducedwithoutendotoxinremoval

生产API而未除内毒素API&excipientsfromnaturalsources

来自自然源的API&赋形剂Manufacturingequipment

生产设备Anywheregram(-)microbesaregrowingorhavebeengrowing

革兰氏阴性细菌生长或曾生长的任何地方39SourcesofPyrogeninParent3940Depyrogenation

去热原AprocesstoremoveordestroyEndotoxin除去或破坏内毒素的一种工艺3logreductionperUSP<1211>

按照USP<1211>，3对数减少值Depyrogenationrequires:

去热原要求：Distillationorultrafiltrationofwater

对水进行蒸馏或超滤Dryheatincineration干热焚烧Destructionbystrongalkalioroxidationwithperoxide用强碱或过氧化物进行氧化来破坏40Depyrogenation

去热原Aprocess4041TypesofDryHeatSterilizers

干热灭菌器的种类Common

常用Forced-convectionbatchsterilizers:utensils,glassware,stainlesssteelequipment,product

强制对流批灭菌器：器具，玻璃器皿，不锈钢设备，产品Tunnel:glassware烘箱：玻璃器皿Flame:ampoulesealing火焰：安瓿封口Lesscommon不常用Microwave微波Laserplasma激光等离子体41TypesofDryHeatSterilizer4142ComponentsforAsepticProcessing

无菌工艺元件Component部件Endotoxin内毒素Method方法LRV*Water水Mostlikely很可能Distillation,RO蒸馏，RO>4Lg.Containers容器Possible可能Chemicals,CIP化学品，CIP>2Excipients赋形剂Possible可能Purification提纯

NDGlassvials玻璃瓶Neverseen未见过Dryheat干热>4Closures塞子Unlikely不可能Wash/sterilize清洗/灭菌>3Lubricant润滑油Neverseen未见过Dryheat干热ND42ComponentsforAsepticProce4243DryHeatDepyrogenation:Glass

干热除热原：玻璃Temp.range:200-350oC

温度范围：200-350oCHeattolerantmaterials,glassware,metalparts,scissors,oils,inorganicsalts

耐热材料，玻璃器皿，金属部件，剪刀，油，无机盐Process:heated,HEPAfilteredair,uniformlydistributedbyconvection&fan

工艺：加热，HEPA过滤空气，通过对流&风扇进行均匀分布Lethaleffect:oxidationofproteinsanddenaturation

致命效应：蛋白质氧化和变性USP<1211>:>3logreduction

大于3个对数减少值43DryHeatDepyrogenation:Gla4344AdvantagesofDry-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

44AdvantagesofDry-heatDepyr4445DisadvantagesofDry-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45DisadvantagesofDry-heatDe4546DepyrogenationbyTunnels

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

46DepyrogenationbyTunnels

通4647ValidationofDepyrogenationCycles

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47ValidationofDepyrogenation4748CaseStudy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Distributionthermocoupletemperaturesmustbewithin±15oCofthesetpointtemperature,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48CaseStudy实例分析

DryHeatDep4849CaseStudy实例分析

DryHeatDepyrogenationTunnel

干热去热原隧道烘箱

ValidationAcceptanceCriteria验证验收标准ThecontrollingRTD(hotzoneEntranceRTD)andthethermocoupleadjacenttothecontrollingRTDmustbewithin±10oCofeachotherduringtheexposure/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±10°C

Aminimumof20of26thermocouplesmustbefunctionalandmeetpostcyclecalibrationcheckcriteriaperXXXXXSOP.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。49CaseStudy实例分析

DryHeatDep4950OtherSterilizationMethods

其他灭菌方法EthyleneOxideSterilization氧化乙烯灭菌Agaseouschemicalagent一种气态化学剂Modeofactionisalkylationofnucleicacid

作用模式为烷化核酸Usedprimarilyfordevices主要用于医疗器材(或设备)Effectiveagainstmostmicroorganismsincludingviruses对大多数微生物(包括病毒)都有效Inexpensive便宜Compatiblewithmostmaterials与大多数物料兼容Disadvantages:toxic,carcinogenic;explosivewithoxygen缺点：有毒，可致癌；与氧接触可能爆炸50OtherSterilizationMethods

5051OtherSterilizationMethods

其他灭菌方法Radiation辐射Usedprimarilyfordevices主要用于医疗器材(或设备)ModeofactionisbreakageofDNAstrands

行动模式为DNA链断裂Gamma(photons)

ɣ-射线（光子）Electronbeam(electrons)电子束（电子）X-ray

X-射线Filtration过滤Tosterilizeheatsensitiveproductsandremovesubvisibleparticlesbysizeexclusionand/oradsorption

对热敏感产品进行灭菌并通过体积排斥和/或吸附法除去可见颗粒Others:EObagsystems,hydrogenperoxide,plasma,ozone,chlorinedioxide,UV,infrared,microwave

其他:EO包装系统，过氧化氢，等离子，臭氧，二氧化氯，UV，红外线，微波51OtherSterilizationMethods

5152Summary

总结Sterileproductposseshighrisk.CGMPshavetobeinplaceallthetime

无菌产品有高风险，CGMP应一直在线。Sterilizationequipmentneedstobequalified.Sterilizationprocesshastobevalidatedwithmicrobiologicalchallenge

灭菌设备应合格。灭菌工艺应通过微生物挑战来验证SALshouldbe1in106orless

SAL应1/106

或更少D,Z,andF0arerequiredtodetermineSAL

要求用D,Z和F0值来确定SALBioburdenhastobecharacterized.应说明生物负荷52Summary

总结Sterileproductp525353ImportantComponentsinSterileProductManufacturing

无菌生产中的重要组成部分PartIIIMethodsofsterilization灭菌方法

SterileProductionandvalidation

无菌生产和验证TheTrendofSterileProduction

无菌产品生产趋势5353ImportantComponentsinSt535454Sterilization

灭菌AGeneralDiscussionfromCGMPPerspective从CGMP角度的综合讨论11Sterilization

灭菌545555Outline

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22Outline讨论纲要PARTI5556PresentationOutline

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3PresentationOutline

概要USReg5657CodeFederalRegulation

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