滅菌的方法及注意事項課件

滅菌的方法及注意事項22Outline

討論綱要PARTITerminology專業(yè)用詞的定義GMPRequirementsinTheManufactureofSterilePharmaceuticalProducts

無菌藥品生產(chǎn)過程中GMP的基本要求PARTIIFacilityDesign生產(chǎn)設(shè)施的設(shè)計HVACSystem空調(diào)系統(tǒng)EnvironmentalMonitoring(EM)環(huán)境的監(jiān)視PharmaceuticalWater制藥用水Cleaning/Sanitation清潔消毒Personnel無菌區(qū)操作人員PARTIIIMethodsofsterilization滅菌方法

SterileProductionandvalidation無菌生產(chǎn)和驗證TheTrendofSterileProduction無菌產(chǎn)品生產(chǎn)趨勢3PresentationOutline

概要USRegulations美國法規(guī)MoistHeatSterilization濕熱滅菌DryHeat/Depyrogenation干熱/去熱原法SterilizationProcessValidation滅菌工藝驗證OtherSterilizationMethods其他滅菌方法4CodeFederalRegulation

美國聯(lián)邦法規(guī)211.84(c)(3)sterileequipment滅菌設(shè)備Sterileequipmentandasepticsamplingtechniquesshallbeusedwhennecessary

必要時應(yīng)使用滅菌設(shè)備和無菌取樣技術(shù)。211.94(c)and(d)sterilized(c)Drugproductcontainersandclosuresshallbecleanand,whereindicatedbythenatureofthedrug,sterilizedandprocessedtoremovepyrogenicpropertiestoassurethattheyaresuitablefortheirintendeduse.

藥品容器和密封系統(tǒng)應(yīng)清潔并根據(jù)藥品的性質(zhì)和要求，進行滅菌，除熱原過程以確保預(yù)期的用途。(d)Standardsorspecifications,methodsoftesting,and,whereindicated,methodsofcleaning,sterilizing,andprocessingtoremovepyrogenicpropertiesshallbewrittenandfollowedfordrugproductcontainersandclosures.

應(yīng)建立并執(zhí)行對藥品容器和密封系統(tǒng)的規(guī)格或質(zhì)量標準，測試方法，清潔方法，滅菌和除熱原過程的相關(guān)書面程序。5CodeFederalRegulation

美國聯(lián)邦法規(guī)211.113(a)and(b)sterilization滅菌(a)Appropriatewrittenprocedures,designedtopreventobjectionablemicroorganismsindrugproductsnotrequiredtobesterile,shallbeestablishedandfollowed.應(yīng)該建立并執(zhí)行用于防止非無菌藥品被致病菌污染的相關(guān)書面程序。(b)Appropriatewrittenprocedures,designedtopreventmicrobiologicalcontaminationofdrugproductspurportingtobesterile,shallbeestablishedandfollowed.Suchproceduresshallincludevalidationofanysterilizationprocess.

應(yīng)該建立并執(zhí)行用于阻止無菌藥品被致病菌污染的相關(guān)書面程序。這些程序應(yīng)包括任何無菌工藝的驗證。6CodeFederalRegulation

美國聯(lián)邦法規(guī)211.167(a)testing（a）測試Foreachbatchofdrugproductpurportingtobesterileand/orpyrogen-free,thereshallbeappropriatelaboratorytestingtodetermineconformancetosuchrequirements.Thetestproceduresshallbeinwritingandshallbefollowed.

對無菌和/或無熱原的每批藥品應(yīng)進行相應(yīng)的實驗室測試以確定其與符合要求。測試程序應(yīng)有書面文件并遵照執(zhí)行。7SterilizationMethods

滅菌方法MoistHeat:

濕熱 Fordrugsanddevices.Themodeofactionisproteindenaturation.

用于藥品和設(shè)備。作用方式是使蛋白質(zhì)變性DryHeat:

干熱 Fordepyrogenationandequipmentsterilization.Themodeofactionisproteindenaturation

用于去熱原和設(shè)備滅菌。作用方式是使蛋白質(zhì)變性EthyleneOxide:

氧化乙烯 Primarilyfordevices.Themodeofactionisalkylationofnucleicacids.主要用于設(shè)備。作用方式是使核酸烷基化Radiation:

輻射 Primarilyfordevices.ThemodeofactionisDNAstrandbreakage主要用于設(shè)備。作用方式是使DNA鏈破壞Othermethods?其他方法？Acceptabletestresultsfornon-condensablegases,super-heatedsteamanddrynessshouldbeobtainedforthecleansteamusedfortheautoclave/SIPCertificationofHEPAfiltrationontheairusedtobreakvacuumorintegritytestingoftheventfilterEmptyvesselheatdistribution空容器熱分布Z-value

Z值作用方式是使核酸烷基化5oC，每分鐘或間隔更短時間記錄數(shù)據(jù)Aminimum3-logreductionofendotoxinmustbedemonstratedforeachendotoxinspikedvialexposedtothedepyrogenationcycle.Gamma(photons)?-射線（光子）0°Cofeachotherduringthestabilizedsterilizationperiod.RunengineeringtrialstofindcoldspotsSummary

總結(jié)耐熱材料，玻璃器皿，金屬部件，剪刀，油，無機鹽Emptyvesselheatdistribution空容器熱分布USRegulations美國法規(guī)Effectiveagainstmostmicroorganismsincludingviruses對大多數(shù)微生物(包括病毒)都有效Sterileproductposseshighrisk.PlaceBIwithin把BI放進無菌灌裝線上設(shè)備和配件的滅菌Ampoules安瓶Z=-1/slopeofthethermalresistancecurve8Bioburden

生物負荷Definition:定義

Populationofviablemicroorganismsonorinaproductand/orapackage

產(chǎn)品和/包裝上的活性微生物的數(shù)量和類型Mixtureoforganisms有機物混合

Indigenousmicroflora地方微植物群

Needstobeinactivatedbysterilization

需滅菌失活9Bioburden

生物負荷BioburdenSources

生物負荷來源Environment環(huán)境 Productcontactsurface,personnel,air

產(chǎn)品接觸表面，人員，空氣Materials材料

Water,rawmaterials.plastic,paper

水，原材料，塑料，紙張CharacteristicsofBioburden生物負荷特點Typesofmicroorganisms微生物類型Resistancetosterilizationprocess對無菌工藝的耐受Numberoforganisms有機物數(shù)量10BiologicalIndicator(BI)

生物指示劑Microbiologicaltestsystemprovidingadefinedresistancetoaspecificsterilizationprocess

微生物測試系統(tǒng)對指定滅菌工藝有明確抵抗性。Acharacterizedpreparationofspecificmicroorganismsresistanttoaparticularsterilizationprocess

某一確定的微生物(指示劑)應(yīng)具有對某一特定滅菌工藝的抵抗性11TypicalBiologicalIndicators

典型生物指示劑Moistheatsterilization

濕熱滅菌GeobacillusstearothermophilusBacillusstearothermophilusDryheatandEOsterilization干熱和環(huán)氧乙烷滅菌BacillusatrophaeusBacillussubtilisvar.niger12FormsofBiologicalIndicators

生物指示劑形式Strips/discsinglassineenvelopes

在透明紙信封里的條形板/光盤Strips/discs

條/光盤Self-contained

獨立包裝的Ampoules安瓶Stripswithmedium中號條形板Liquidsuspension

液體懸浮液Metal

金屬Fiberglass

玻璃纖維13ChoiceofanAppropriateBI

相關(guān)生物指示劑的選擇Sterilizationprocess

滅菌工藝Cycledesignmethod

循環(huán)設(shè)計方法Productbioburden

產(chǎn)品生物負荷Population數(shù)量Resistance抵抗性14BIsPreparedbyUser

生物指示劑的準備Performance

性能Resistance抵抗性Population數(shù)量Purity純度D-

value

D值Recoverymethods恢復(fù)方法Storagerequirements儲存要求15BiologicalIndicatorUse

生物指示劑使用PlaceBIwithin把BI放進Product產(chǎn)品Package包裝Sterilizerloadtomonitorprocess

滅菌器負荷以監(jiān)視滅菌工藝Exposetosterilizingconditions暴露在滅菌狀態(tài)RemoveBIandtest移除BI和測試Countsurvivors生存數(shù)量的計算Growth/nogrowthresponse生長/無生長反應(yīng)16D-value

D值TheDvalueisthetime,usuallyinminutes,requiredtoachieveinactivationof90%(oronelogarithm)ofapopulationofthetestmicroorganismatspecifiedconditions.

D值是在特定條件下微生物數(shù)量降低的90%（或一對數(shù)）所用的時間，通常是以分鐘為單位的。

BacillusstearothermophilushasaDvalue:

嗜熱脂肪芽胞桿菌的D值2 min at121oC20 min at110oC0.2-0.3min at130oC

Ofalltheaspectsofsterilizationvalidation,theDvalueisperhapsthemostimportant.ValidatingaprocesswithoutconsiderationoftheDvalueislargelyineffectiveandisnotacceptablefromCGMPperspective

滅菌驗證中，D值可能是最重要的。不考慮D值的驗證過程多半是無效的且不被CGMP接受。17Z-value

Z值Z-value:numberofdegreesoftemperaturerequiredfora1logarithmchangeintheD-value

Z值：D值1對數(shù)改變需要的溫度數(shù)Z=-1/slopeofthethermalresistancecurve

Z=-1/熱阻力曲線斜率

where Slope=logarithmicchangeinD-value/changeintemperature

斜率=

D值上對數(shù)的改變/溫度的改變18ImpactofZ-value

Z值的影響Whenz-valueissmall,considerablylessinactivationwillresultbelowreferencetemperatureandconsiderablymoreabovethereferencetemperature

當(dāng)Z值較小時，較低程度的失活將導(dǎo)致溫度低于參考溫度，較大程度的失活將導(dǎo)致溫度高于參考溫度。MaterialsdryattheendofcycleAtleast10TCsof12thermocouplesmustbefunctionalattheendoftherunManufacturingequipment(a)Appropriatewrittenprocedures,designedtopreventobjectionablemicroorganismsindrugproductsnotrequiredtobesterile,shallbeestablishedandfollowed.生產(chǎn)API而未除內(nèi)毒素作用方式是使蛋白質(zhì)變性Ampoules安瓶Indigenousmicroflora地方微植物群Recordingdevicecapableofsupporting≥12tempsensorswithanaccuracyof≥0.ValidationAcceptanceCriteria驗證驗收標準最終任何傳感器的失敗應(yīng)調(diào)查研究。Performance性能(c)Drugproductcontainersandclosuresshallbecleanand,whereindicatedbythenatureofthedrug,sterilizedandprocessedtoremovepyrogenicpropertiestoassurethattheyaresuitablefortheirintendeduse.Failuretomeetacceptancecriteriashallrequirecycleparametermodifications,loadpatternmodificationand/orequipmentrepairs/correctionsProcureorprepareendotoxinindicators受控并記錄的3次成功運行Effectiveagainstmostmicroorganismsincludingviruses對大多數(shù)微生物(包括病毒)都有效用于藥品和設(shè)備。作用方式是使DNA鏈破壞SterilizationProcessValidation滅菌工藝驗證ThedrainPT-100andTCinthedrainmustbewithin1.19TypicalTemperatureProfiles

典型溫度分布20TypeofSterilization

滅菌類型MoistHeatSterilization濕熱滅菌21MoistHeatSterilization

濕熱滅菌Characteristics:

特征Wellunderstoodandwellcharacterizedprocess,firstvalidatedprocessinpharmindustry

濕熱滅菌是一已被很好地理解并描述滅菌工藝，也是在制藥工業(yè)中第一個被驗證的滅菌工藝Suitableforawidevarietyofapplications

適合于較大應(yīng)用范圍Equipmentisreadilyavailable

設(shè)備很容易從市場購買到Costonaperusebasisislow

每次使用基準花費低22MoistHeatSterilization

濕熱滅菌Applications:

應(yīng)用Terminalsterilizationofparentalproduct

注射劑的終端滅菌Sterilizationofequipmentandcomponentsforuseinasepticfilling

無菌灌裝線上設(shè)備和配件的滅菌Sterilizationoflaboratorymaterials

實驗室用材料的滅菌In-situsterilizationofprocesspipingandequipment(SIP)工藝管道和設(shè)備在線滅菌

23BasicTypesofMoistHeatSterilization

濕熱滅菌基本類型Saturatedsteam飽和蒸汽Autoclaves(self-closing)

高壓滅菌柜（半封閉）SIP在線滅菌Superheatedwater過熱水Spray

噴霧Submerged

浸沒的SIP

在線滅菌Steam-air-mixture(SAM)

水蒸氣－空氣混合物24BasicElementsofSterilizationProcessValidation

滅菌工藝驗證的基本元素Emptyvesselheatdistribution空容器熱分布Heatdistributionandpenetration熱分布和滲透3. Microbiologicalchallenges

微生物挑戰(zhàn)25SteamSterilizationValidation:Prerequisites

蒸汽滅菌驗證：前提OQforanautoclave:高壓滅菌柜運行確認Emptychambertemperaturemappingwithin±1.0oCofthemean空腔體溫度分布圖在平均值的±1.0oC內(nèi)Chamberintegritytest(noleaking)腔體完整性測試CertificationofHEPAfiltrationontheairusedtobreakvacuumorintegritytestingoftheventfilter

用于隔斷真空或通氣過濾器完整性測試的空氣HEPA過濾認證RequirementsforSIP在線滅菌要求Temperaturemapping溫度分布圖Anintegritytest,whereappropriate相關(guān)完整性測試UseofBI生物指示劑的使用Allcriticalinstrumentsmustbecalibrated

所有關(guān)鍵儀器需校驗26SteamSterilizationValidation:Prerequisites

蒸汽滅菌驗證：前提Acceptabletestresultsfornon-condensablegases,super-heatedsteamanddrynessshouldbeobtainedforthecleansteamusedfortheautoclave/SIP

應(yīng)獲得用于高壓滅菌柜/SIP的潔凈蒸汽中的不凝氣體，過熱蒸汽及干燥度的可接受測試結(jié)果5.ToolsfortheconductofthePQstudy:

進行PQ研究的工具BIwith106sporesandknownDandZvalues

BI有106個孢子，已知D值和Z值Temperaturesensors溫度傳感器Recordingdevicecapableofsupporting≥12tempsensorswithanaccuracyof≥0.5oC,recordingdataeveryminuteorless

記錄設(shè)備能支撐≥12溫度傳感器，精度≥0.5oC，每分鐘或間隔更短時間記錄數(shù)據(jù)Meansofintroducingtempsensorsintotheautoclave/SIP將溫度傳感器導(dǎo)入高壓滅菌柜/SIP的方法27SteamSterilizationValidation:PreparationofPQprotocol

蒸汽滅菌驗證：PQ方案的準備Aprotocolshallbepreparedfor:

對于下列各項應(yīng)建立方案：Newautoclave/SIP新高壓滅菌柜/SIPNewloadingpatternsorproductconfigurations

新裝料方式或產(chǎn)品配置Changestoexistingpatterns

對現(xiàn)有裝料方式的變更Changestooperationcycleparameters

對運行周期參數(shù)的變更Majorchangetoequipmentasdirectedbychangecontrol變更控制要求的設(shè)備主要變更28SteamSterilizationValidation:PreparationofPQprotocol

蒸汽滅菌驗證：PQ方案的準備Theprotocolmayinclude:方案可能包括：Objectivesofthevalidationstudy

驗證研究的目的Identificationanddescriptionofthesterilizeranditsprocesscontrols滅菌器的識別和說明及工藝控制IdentificationofSOPsfortheprocessequipment

工藝設(shè)備SOP的識別DescriptionoforSOPreferenceforinstrumentcalibrationprocedures儀器校驗程序的說明或SOP參考Identificationofcalibrationproceduresfortemp-monitoringequipment,whichincludeatwopointpre-runcalibrationandapost-runverificationforeachrun

溫度監(jiān)測設(shè)備校驗程序的識別，包括一個兩點預(yù)運行校驗和每次運行后的確認Processparameteracceptancecriteria

工藝參數(shù)的驗收標準Majorchangetoequipmentasdirectedbychangecontrol變更控制要求的設(shè)備主要變更TheautoclaveshallbeloadedandoperatedinaccordancewithSOP高壓滅菌器應(yīng)按照SOP進行裝載并運行工藝：加熱，HEPA過濾空氣，通過對流&風(fēng)扇進行均勻分布Compatiblewithmostmaterials與大多數(shù)物料兼容無菌灌裝線上設(shè)備和配件的滅菌在穩(wěn)定滅菌期間，排水管PT-100和排水管中TC之間溫差必須在1.GlassvialsObjectivesofthevalidationstudy驗證研究的目的Othermethods?其他方法？BIwith106sporesandknownDandZvaluesF0mustbenotlessthanpresettimeof15minutesF0不得低于預(yù)設(shè)定的15分鐘PlaceBIwithin把BI放進Aminimum3-logreductionofendotoxinmustbedemonstratedforeachendotoxinspikedvialexposedtothedepyrogenationcycle.PharmaceuticalWater制藥用水NLT12sensorsforautoclave這些程序應(yīng)包括任何無菌工藝的驗證。Heatdegradationlimitsmaterials熱降解限制材料84(c)(3)sterileequipment滅菌設(shè)備Environmental:complexofprotein,carbohydrate&lipid.USRegulations美國法規(guī)Cleaning/Sanitation清潔消毒ComponentsforAsepticProcessing

無菌工藝元件APIsproducedwithoutendotoxinremovalPharmaceuticalWater制藥用水Self-contained獨立包裝的RunengineeringtrialstofindcoldspotsForheatpenetrationdata,belocatedsuchthattheypenetratetheequipment,ortheproductcontainerandareimmersedintheproduct對于熱穿透數(shù)據(jù)，應(yīng)置于能穿透設(shè)備或產(chǎn)品容器處，并包含在產(chǎn)品中PresentationOutline

概要TerminologyandDefinitions

術(shù)語和定義Themodeofactionisproteindenaturation.Distillation,RO蒸餾，ROSterilizationoflaboratorymaterialsSuchproceduresshallincludevalidationofanysterilizationprocess.Distillationorultrafiltrationofwater帶孔玻璃瓶3個對數(shù)減少值，單位為EU(c)Drugproductcontainersandclosuresshallbecleanand,whereindicatedbythenatureofthedrug,sterilizedandprocessedtoremovepyrogenicpropertiestoassurethattheyaresuitablefortheirintendeduse.對現(xiàn)有裝料方式的變更行動模式為DNA鏈斷裂1oC作為基底溫度，10oC作為Z值，F(xiàn)o應(yīng)為計算值Thetemperaturemeasuredinanyfluidcontainersisnotgreaterthan80oCattheendofthecycle循環(huán)結(jié)束時，任何液體容器中測量的溫度不超過80oCIfthetempcontrollingsensorisnotinthedrain,anadditionalsensorshallbeplacedinthedrain如果溫度控制傳感器不在排水口中，應(yīng)置另一個傳感器于排水口中Populationofviablemicroorganismsonorinaproductand/orapackage29SteamSterilizationValidation:PreparationofPQprotocol

蒸汽滅菌驗證：PQ方案的準備Adescriptionofthefollowing:以下說明Biodurdendeterminationstudies生物負荷確認研究Emptychamberheatdistributionstudies(±1oC)

空腔體熱分布研究（±1oC）Loadedchamber(includingLoadconfiguration,maxladingandminloading)heatpenetrationstudies

滿載腔體（包括裝載配置,最大和最少）熱穿透研究Containermappingstudies(maynotneededif<100mL)

容器分布圖研究（如果容量<100mL，可能不需要）Microbiologicalchallengestudies微生物挑戰(zhàn)研究Evaluationofdrugproductcoolingwater(whereapplicable)

藥品冷卻水評估（若適用）Evaluationofventfiltermembranesassociatedwiththesterilizer與滅菌器相關(guān)的通氣過濾膜的評估30SteamSterilizationValidation:PreparationofPQprotocol

蒸汽滅菌驗證：PQ方案的準備4. Descriptionofthetempsensorplacementwithintheload在負荷范圍內(nèi)溫度傳感器位置的說明Onenexttothetempcontrollingsensorforautoclave.一1個接近高壓滅菌柜的溫度控制傳感器NLT12sensorsforautoclave

不少于12個傳感器用于高壓滅菌柜NLT3sensorsforSIP不少于3傳感器用于在線滅菌Ifthetempcontrollingsensorisnotinthedrain,anadditionalsensorshallbeplacedinthedrain

如果溫度控制傳感器不在排水口中，應(yīng)置另一個傳感器于排水口中Aminimum#ofsensorsmeetingthecalibrationandverificationshallbeestablished.Anysensorthatfailattheendofthestudyshallbeinvestigated.Anysensorlocatednexttothetempcontrollingsensororinthedrainshallmeetthepreandpostcalibrationaspartoftheacceptancecriteriaforasuccessfulrun應(yīng)建立符合校驗和驗證的最少#傳感器。最終任何傳感器的失敗應(yīng)調(diào)查研究。任何臨近溫度控制傳感器或在排水口的傳感器應(yīng)符合預(yù)校驗和后校驗，這是成功運行的驗收標準的一部分。31SteamSterilizationValidation:PreparationofPQprotocol

蒸汽滅菌驗證：PQ方案的準備5. Acceptancecriteriafortemp.pressure,Fowhereappropriate,vacuumanddestructionsofBIs

溫度壓力，相關(guān)Fo，真空及生物指示劑滅活的驗收標準6. Arequirementfor3consecutive,successfulruns

3次連續(xù)成功運行的要求

Thedrainisthecoldestpointandisoutsideofthesterilizationzone,notincludedinthecalculationofmeanchambertemperatureandnotsubjecttochamberdistributionrequirement.

排水口是最冷點，在滅菌區(qū)外，且不包括在平均腔體溫度計算內(nèi)，不受限于腔體分布要求

Thermocouplesshouldbeplacedatthesamelocationforemptychamberandloadedchamberheatdistributionstudies.

熱電偶應(yīng)置于相同位置，進行空腔體和滿載腔體熱分布研究

Uniformityisexpectedonlyatthesteadystate.

僅在穩(wěn)定狀態(tài)下要求均一性32SteamSterilizationValidation:ExecutionofPQ

蒸汽滅菌驗證：PQ執(zhí)行1. Temperaturesensorshall:溫度傳感器應(yīng)：Forheatpenetrationdata,belocatedsuchthattheypenetratetheequipment,ortheproductcontainerandareimmersedintheproduct對于熱穿透數(shù)據(jù)，應(yīng)置于能穿透設(shè)備或產(chǎn)品容器處，并包含在產(chǎn)品中Forheatdistributiondata,beevenlydistributedwithintheloadorsystemandshallnotcontactmetalsurface

對于熱分布數(shù)據(jù)，平均分布在負荷范圍內(nèi)或系統(tǒng)中，且不能接觸金屬面2. TheBIshall:生物指示劑應(yīng)：Beplacednearthetipofthetemperaturesensorusedforpenetration置于靠近用于穿透的溫度傳感器的頂端IncludetheidentificationthatlinkstothenumberoftheappropriatetemperaturesensoroftheBI

包括與相關(guān)BI溫度傳感器數(shù)量有關(guān)的識別TheDvalueoftheBIshallbedeterminedintheproduct

BI的D值應(yīng)在生產(chǎn)中確定33SteamSterilizationValidation:

ExecutionofPQ

蒸汽滅菌驗證：PQ執(zhí)行TheautoclaveshallbeloadedandoperatedinaccordancewithSOP高壓滅菌器應(yīng)按照SOP進行裝載并運行Using121.1oCasthebasetemperatureand10oCastheZ-value,Foshallbecalculated121.1oC作為基底溫度，10oC作為Z值，F(xiàn)o應(yīng)為計算值A(chǔ)cceptancecriteriashallbebasedonnogrowthonexposedBIsandgrowthonthepositivecontrol

BI驗收標準應(yīng)依據(jù)暴露的BI上未生長和陽性控制BI上的生長Failuretomeetacceptancecriteriashallrequirecycleparametermodifications,loadpatternmodificationand/orequipmentrepairs/corrections

未達到驗收標準應(yīng)要求周期參數(shù)變更，負荷曲線圖變更和/或設(shè)備修理/修正34CaseStudy實例分析

AutoclaveSterilizationValidation

高壓蒸汽滅菌驗證Table1 AcceptanceCriteriaofEmptyChamberHeatDistribution表1空腔體熱分布驗收標準No.AcceptanceCriteria驗收標準1The12TCsdistributedthroughoutthechambermustbewithin±1oCofthemeanthermocoupletemperatureeachminuteafterstabilizationthroughoutthedurationofthesterilizationphase.整個滅菌階段，穩(wěn)定后，每分鐘分布在腔體內(nèi)的12個TC必須在平均熱電偶溫度的±1oC

2Dwelltimenotlessthanthepre-setsterilizationtimeof40minutes.停留時間不少于預(yù)設(shè)定的滅菌時間40分鐘3TheTCsmonitoringthedrainandchamberRTDsmaynotfailpostcheckcalibration.TenofthetwelveTCsmustbefunctionalattheendofthecycle.監(jiān)測排水和腔體RTD的TC不應(yīng)在實驗后校驗失敗。循環(huán)結(jié)束時，12個TC中的10個必須在實驗后校驗時功能完好35CaseStudy

實例分析

AutoclaveSterilizationValidation

高壓蒸汽滅菌驗證Table2

AcceptanceCriteriaforHeatDistribution表2熱分布驗收標準No.AcceptanceCriteria驗收標準1Dwelltimemustbenotlessthanthepre-setsterilizationtimeof15minutes停留時間不得少于預(yù)設(shè)定的滅菌時間15分鐘2ThedrainPT-100andTCinthedrainmustbewithin1.0°Cofeachotherduringthestabilizedsterilizationperiod.在穩(wěn)定滅菌期間，排水管PT-100和排水管中TC之間溫差必須在1.0oC之內(nèi)3Thetemperaturedeviationduringholdingtimesdoesnotexceed-1°Cand+2°Cfromthesetpoint(duringthestabilizationperiod)停留時間的溫度偏差不超過-1oC及設(shè)定點的+2oC（在穩(wěn)定階段）4Atleast10TCsof12thermocouplesmustbefunctionalattheendoftherun循環(huán)結(jié)束時，12個TC中的10個必須功能完好36CaseStudy實例分析

AutoclaveSterilizationValidation

高壓蒸汽滅菌驗證Table3

AcceptanceCriteriaforHeatPenetration表3熱分布驗收標準No.AcceptanceCriteria驗收標準1F0mustbenotlessthanpresettimeof15minutesF0不得低于預(yù)設(shè)定的15分鐘2ThedrainPT-100andTCinthedrainmustbewithin1.0°Cofeachotherduringthestabilizedsterilizationperiod.在穩(wěn)定滅菌期間，排水管PT-100和排水管中TC之間溫差必須在1.0°C之內(nèi)3Thetemperaturesensorsremainedinpositionattheendofcycle循環(huán)結(jié)束時溫度傳感器仍然在位4Theitemscontainingsensorsareintact;bottleshavenotleaked,burstorbroken裝傳感器的器具完整；瓶子不漏，爆裂或破碎5Nomorethan1%ofthebottleshasburstorbroken爆裂或破碎的瓶不超過1%6Thetemperaturemeasuredinanyfluidcontainersisnotgreaterthan80oCattheendofthecycle循環(huán)結(jié)束時，任何液體容器中測量的溫度不超過80oC7Thetemperaturedeviationduringholdingtimedoesnotexceed-1°Cand+2°Cfromthesetpointduringthestabilizedsterilizationperiod停留時間的溫度偏差不超過-1°C及穩(wěn)定滅菌階段設(shè)定點的+2°C8Nosporesurvivalofthebiologicalindicators生物指示器上無孢子生存9Atleast10TCsof12thermocouplesarefunctionalattheendoftherunforthemaximum&minimumloadconfigurations,respectively每次最大&最小裝載配置結(jié)束時，12個熱電偶中至少10個TC起作用10Autoclaveandvalidationtimersmustbesynchronized高壓滅菌柜和確認計時器必須同步37DryHeatSterilization干熱滅菌

Depyrogenation去熱原38TerminologyandDefinitions

術(shù)語和定義Pyrogen:Feverproducingsubstance:endotoxin,viruses,fungi,toxinfromgram(+)andgram(-)bacterials,peptiglycan

熱原：熱產(chǎn)生的物質(zhì)：內(nèi)毒素，病毒，真菌，革蘭氏陽性和陰性細菌產(chǎn)生的毒素，肽糖Endotoxin:Componentfromoutermembraneofgram(-)bacterial,mostpotentpyrogens,LPSinstructure

內(nèi)毒素：革蘭氏陰性細菌外膜的成分，大多數(shù)有效熱原，結(jié)構(gòu)中的LPSEnvironmental:complexofprotein,carbohydrate&lipid.

環(huán)境：復(fù)合蛋白質(zhì)，碳水化合物&脂類

Purified:lipopolysacchride,usedinendotoxinstandards

提純后：脂多糖，用于內(nèi)毒素標準39SourcesofPyrogeninParenterals

注射用藥熱原的來源Watersystemsandresincolumns

水系統(tǒng)和樹脂交換柱APIsproducedwithoutendotoxinremoval

生產(chǎn)API而未除內(nèi)毒素API&excipientsfromnaturalsources

來自自然源的API&賦形劑Manufacturingequipment

生產(chǎn)設(shè)備Anywheregram(-)microbesaregrowingorhavebeengrowing

革蘭氏陰性細菌生長或曾生長的任何地方40Depyrogenation

去熱原AprocesstoremoveordestroyEndotoxin除去或破壞內(nèi)毒素的一種工藝3logreductionperUSP<1211>

按照USP<1211>，3對數(shù)減少值Depyrogenationrequires:

去熱原要求：Distillationorultrafiltrationofwater

對水進行蒸餾或超濾Dryheatincineration干熱焚燒Destructionbystrongalkalioroxidationwithperoxide用強堿或過氧化物進行氧化來破壞41TypesofDryHeatSterilizers

干熱滅菌器的種類Common

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

強制對流批滅菌器：器具，玻璃器皿，不銹鋼設(shè)備，產(chǎn)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激光等離子體42ComponentsforAsepticProcessing

無菌工藝元件Component部件Endotoxin內(nèi)毒素Method方法LRV*Water水Mostlikely很可能Distillation,RO蒸餾，RO>4Lg.Containers容器Possible可能Chemicals,CIP化學(xué)品，CIP>2Excipients賦形劑Possible可能Purification提純

NDGlassvials玻璃瓶Neverseen未見過Dryheat干熱>4Closures塞子Unlikely不可能Wash/sterilize清洗/滅菌>3Lubricant潤滑油Neverseen未見過Dryheat干熱ND43DryHeatDepyrogenation:Glass

干熱除熱原：玻璃Temp.range:200-350oC

溫度范圍：200-350oCHeattolerantmaterials,glassware,metalparts,scissors,oils,inorganicsalts

耐熱材料，玻璃器皿，金屬部件，剪刀，油，無機鹽Process:heated,HEPAfilteredair,uniformlydistributedbyconvection&fan

工藝：加熱，HEPA過濾空氣，通過對流&風(fēng)扇進行均勻分布Lethaleffect:oxidationofproteinsanddenaturation

致命效應(yīng)：蛋白質(zhì)氧化和變性USP<1211>:>3logreduction

大于3個對數(shù)減少值用于隔斷真空或通氣過濾器完整性測試的空氣HEPA過濾認證帶孔玻璃瓶3個對數(shù)減少值，單位為EUNeedstobeinactivatedbysterilizationSterilizationoflaboratorymaterialsAPIsproducedwithoutendotoxinremovalSterilizationofequipmentandcomponentsforuseinasepticfillingPresentationOutline

概要Bioburden

生物負荷BI有106個孢子，已知D值和Z值ComponentsforAsepticProcessing

無菌工藝元件Run3fullloadswithendotoxinindicatorsinplaceatcoldspotsandrandomsitesThermocouplesshouldbeplacedatthesamelocationforemptychamberandloadedchamberheatdistributionstudies.環(huán)境：復(fù)合蛋白質(zhì)，碳水化合物&脂類Thetemperaturemeasuredinanyfluidcontainersisnotgreaterthan80oCattheendofthecycle循環(huán)結(jié)束時，任何液體容器中測量的溫度不超過80oCDepyrogenation

去熱原BIwith106sporesandknownDandZvaluesIfthetempcontrollingsensorisnotinthedrain,anadditionalsensorshallbeplacedinthedrain如果溫度控制傳感器不在排水口中，應(yīng)置另一個傳感器于排水口中FormsofBiologicalIndicators

生物指示劑形式ThedrainPT-100andTCinthedrainmustbewithin1.斜率=D值上對數(shù)的改變/溫度的改變Cleaning/Sanitation清潔消毒Temperaturemapping溫度分布圖44AdvantagesofDry-heatDepyrogenation

干熱去熱原的優(yōu)點Inactivatespyrogenswhilesterilization

滅菌時使熱原失活

Materialsdryattheendofcycle

循環(huán)結(jié)束時物料干燥Corrosiveeffectsareminimal

腐蝕性最小Conveyorsystemsallowforhighertempsandshorterdwelltime

傳送系統(tǒng)允許更高溫度和更短的停留時間

45DisadvantagesofDry-heatDepyrogenation

干熱去熱原的缺點Slowprocess(airisapoorconductor)

降低工藝速度（空氣是不良導(dǎo)體）Heatpenetrationslowerthansteam

熱穿透比蒸汽慢Ratevaries(slowforglass,rapidforstainlesssteel)

速度各異（玻璃慢，不銹鋼快）Heatmustpenetratetoinnersurfaceviaconduction

熱必須通過傳導(dǎo)穿透到內(nèi)表面Layeringcanoccurduetodifferencesinairdensitywithtemperature;mechanicalcirculationneeded

空氣密度，溫度不同可能導(dǎo)致分層，需要機械循環(huán)Heatdegradationlimitsmaterials熱降解限制材料Contractionduringcoolingmaydrawcontaminants

冷卻過程中接觸可能引起污染46DepyrogenationbyTunnels

通過烘箱去熱原HEPA-filtered,verticallaminarairflowinheatingandcoolingzones,orradiantheatersinheatingzoneandverticallaminarairflowincoolingzone

在加熱和冷卻區(qū)經(jīng)HEPA過濾的垂直層流氣流，或加熱區(qū)的輻射加熱器和冷卻區(qū)的垂直層流氣流Conveyorbelttoprovidein-linecontinuousflowofsterileglasswaretoasepticarea

傳送帶提供連續(xù)在線的無菌玻璃器皿到無菌區(qū)的流動Limitedtoonetypeofloadatatime

一次限用一種裝載方式Problemtocontrolspeedmatchotherlineequipment(filler)

控制速度以匹配其他線設(shè)備（灌裝機）的問題Difficulttoachieveuniformheating,andheatsourcemaygenerateparticles

很難達到均勻加熱，熱原可能產(chǎn)生顆粒Largeproductvolumeneededtojustifytunnel

調(diào)整烘箱要求大的產(chǎn)品容量

47ValidationofDepyrogenationCycles

去熱原循環(huán)驗證Runengineeringtrialstofindcoldspots

進行試運行以找到冷點Runtrialstodetermineworsecasevial,e.g.,thickestglass,packingeffects

試車以確定最差狀況的玻璃瓶，如最厚的玻璃，緊束效應(yīng)Procureorprepareendotoxinindicators

獲得或制備內(nèi)毒素指示劑Run3fullloadswithendotoxinindicatorsinplaceatcoldspotsandrandomsites

進行3次滿載運行，內(nèi)毒素指示劑置于冷點和隨機位置Acceptancecriteria驗收標準min.3logreductioninEUforspikedvials

帶孔玻璃瓶3個對數(shù)減少值，單位為EU3successfulrunswithcontrolsanddocumentation

受控并記錄的3次成功運行48CaseStudy實例分析

DryHeatDepyrogenationTunnel

干熱去熱原隧道烘箱

ValidationAcceptanceCriteria驗證驗收標準Aminimumofthreesuccessfulvalidationrunsmustbeperformed.必須至少進行三次成功的驗證Distributionthermocoupletemperaturesmustbewithin±15oCofthesetpointtemperature,afterstabilization.

穩(wěn)定后，分配熱電偶溫度必須在設(shè)定溫度的±15oC內(nèi)AminimumcumulativeFHvalueoftwelveminutesmustbedemonstratedforeachpenetrationthermocoupleattheendofeachcycle.

每次循環(huán)結(jié)束時，必須證明每個滲透熱電偶12分鐘內(nèi)的最少累計FH值A(chǔ)minimum3-logreductionofendotoxinmust