清潔驗證之取樣方法

清潔驗證之取樣方法（PDATR49內(nèi)容節(jié)選5中英文版）5.0SamplingMethods取樣方法Itisessentialtoacleaningvalidationprogramthattheappropriatesamplingtechniquesareutilized.Samplingmustbeconductedwithtechniquesappropriatefortheequipmentsurfacesandforthenatureofthestudy,includingtheanalyticalmethodsused.Thissectiondiscussestypesofsamplingmethods,samplingrecoveryvalidationstudies,andthetrainingandqualificationofsamplers.在清潔消毒驗證中采用適當(dāng)?shù)娜蛹夹g(shù)是必要的。取樣過程中涉及的技術(shù)方法應(yīng)與設(shè)備表面和研究本質(zhì)相適應(yīng)，包括使用的分析方法。這一部分討論了取樣方法的類型，驗證取樣回收研究以及取樣人員的培訓(xùn)及資質(zhì)。SamplingMethodSelection取樣方法選擇Selectionofasamplingmethoddependsonthenatureoftheequipmentandthenatureoftheresiduebeingmeasured.Samplingmethodsdiscussedherearedirectsurfacesampling,swabbing,rinsewatersamplingandplacebosampling.Itshouldbenotedthatwhileregulatorydocumentsrefertoswabbingas“direct”samplingandtorinsewatersamplingas“indirect”sampling,itispreferableandmoredescriptivetorefertothosesamplingmethodsas“swabsampling”and“rinsesampling,”andreservetheterm“directsampling”fortechniquessuchastheuseofvisualinspection.取樣方法的選擇取決于設(shè)備性質(zhì)以及檢測殘留物的性質(zhì)。此處討論的取樣方法是直接表面取樣，擦拭取樣，沖洗水取樣以及空白對照取樣。值得注意的是法規(guī)文件中指出擦拭被認(rèn)為是“直接”取樣，沖洗水取樣被認(rèn)為是“間接”取樣，所指的取樣方法如“擦拭取樣”和“沖洗水取樣”是更可取并且更好被描述的，技術(shù)上保留“直接取樣”的術(shù)語，比如使用目測檢查。DirectSamplingMethods直接取樣方法Directsamplingmethods(asusedinthisdocument)includevisualinspection.直接取樣方法(本文中使用的)包括目測檢查。Itisawell-acceptedpracticethatacleaningprocessshouldremovevisibleresiduesfromtheproductionprocessoffofequipmentsurfaces.Thevisualinspectionofequipmenthaslimitationsinthatsomeequipmentsurfaces(e.g.,piping)areusuallynotaccessibleforviewing.Theuseofopticalequipmentlikemirrorsorendoscopes,aswellastheuseofadditionallighting,canhelptofacilitatevisualinspection.清潔過程中應(yīng)除去設(shè)備表面上來自于生產(chǎn)工藝可見殘留，這一點是被廣泛認(rèn)可的。設(shè)備的目測檢查具有局限性，因為一些設(shè)備表面(比如管道)通常是不易觀察的。像鏡子或者內(nèi)視鏡之類的光學(xué)儀器的使用作為輔助照明能夠幫助進(jìn)行目測檢查。Remoteinspectiontechniques(withfiber-opticprobesandaLCDviewingscreen)areutilizedwhenvisualinspectionbyatrainedinspectorisdifficulttoperformbecauseofaccesstoequipmentsurfaces,orwhenonepreferstosupplementan“unaided”visualinspectionprocedure.當(dāng)由經(jīng)過培訓(xùn)的人員由于要接觸設(shè)備表面而難以進(jìn)行目檢時或者是補充“未受協(xié)助”的目檢程序時，遠(yuǎn)程檢查技術(shù)（纖維光學(xué)探頭以及LCD觀察屏）被應(yīng)用。Borescopes,fiberscopes,andvideoscopesallowvisualinspectionofhard-to-reachareas.Borescopeshavebeenusedtoviewtheinteriorofpipingandtankwelds.Typicalbenefitsofthesescopesarethatthey:canfitintoconfinedspacesnotaccessibletooperators;areverymaneuverable;haveadditionallightingattached;andmaycomewithoptionalmagnificationand/orzoomingcapabilities.Themajordrawbackofthesescopesisthecomplexityofuse,controllinglighting/brightness,andthattheoperatorstillhastomakethedeterminationiftheareaviewedisvisuallyclean.管道內(nèi)窺鏡，纖維內(nèi)窺鏡以及光纖視鏡允許對難以達(dá)到的區(qū)域進(jìn)行目檢。管道內(nèi)窺鏡用于觀察管道內(nèi)部和水槽焊接點。這些范圍的典型好處是：適用于操作人員接觸不到的密閉空間；易操作，具有輔助照明并且具有選擇性放大和/或提升的能力。這些范圍的最大缺點是使用復(fù)雜，控制照明/亮度，并且操作人員需要判斷該區(qū)域是否能夠清晰可見。Aremotevisualcameraallowsoperatorstoviewremoteareasonascreen.Thecamerahasmostofthesamestrengthsandweaknessesasthescopes,withtheaddedbenefitthatoperatorscantypicallyalsorecordvideoortakepictures.Multipleoperatorscan,atthesametime,viewwhatisonthescreen.Thepotentialtorecordvideoandallowmultipleoperatorstoviewthescreenmayhelpsupportasite’svisualinspectiontrainingprogram.Picturesprintedfromthecameramaydistorttheactualamountofresiduepresent,sinceoperatorswilltypicallyzoominonaparticularareawhentakingapicture.遠(yuǎn)程可視相機(jī)允許操作人員在屏幕上觀察遠(yuǎn)程區(qū)域。相機(jī)與范圍具有大部分相同的優(yōu)勢和劣勢，而更多的優(yōu)勢是操作人員能夠代表性的記錄視頻或者拍照。多個操作人員能夠同時觀察到屏幕顯示了什么。記錄視頻以及允許多個觀察者觀察屏幕的可能性可以幫助支持廠區(qū)目檢培訓(xùn)程序。來自相機(jī)的打印的圖片可能扭曲殘留的實際數(shù)量，因為操作人員將在特殊區(qū)域內(nèi)聚焦，當(dāng)拍照時。Allthesetechniques,likevisualinspection,requireanadequatetrainingprogram.所有這些的技術(shù)，像是目檢，要求具有充足的培訓(xùn)程序。RinseSampling沖洗取樣Rinsesamplinginvolvessamplingtheequipmentbyflowingwateroverallrelevantequipmentsurfacestoremoveresidues,whicharethenmeasuredintherinsewater.Themostcommonrinsesamplingtechniqueistotakeagrabsamplefromthefinalrinsewaterduringthefinalrinseofthecleaningprocess.Anotheroptionistofilltheentireequipmentwithwaterafterthecleaningprocedureiscompleted.Then,abulksampleistakenandanalyzed.AthirdoptionistoutilizeaseparateCIPsamplingrinseofdefinedvolumefollowingthecompletionofthefinalprocessrinse.沖洗取樣涉及對設(shè)備取樣通過流水沖洗所有相關(guān)設(shè)備表面移除殘留，殘留將在沖洗水中進(jìn)行測量。大部分沖洗取樣技術(shù)是在清潔過程中的最終沖洗中通過從最終沖洗水中提取隨機(jī)樣品。另一個選擇是在完成清潔程序后，用水填滿整個設(shè)備。然后，取大部分樣品進(jìn)行分析。第三個選擇是使用單獨的CIP取樣沖洗定量，在最終沖洗完成后。AdvantagesanddisadvantagesofbothmethodsforCIPrinsingareshowninTable5.1.2.CIP沖洗的兩種方法的優(yōu)勢和劣勢見表格5.1.2Table5.1.2ComparisonofCIPGrabSamplingversusSeparateCIPSamplingRinse表格5.1.2CIP簡單取樣與單獨CIP取樣沖洗的比較GrabSamplingfromCIPFinalRinse來自CIP最終沖洗的簡單取樣SeparateCIPSamplingRinse單獨的CIP取樣沖洗Advantages優(yōu)勢Representsthenormalcleaningprocess代表了正常的清潔過程Allowson-linetesting允許在線檢測Requiresnoadditionalamountsofrinsewater不要求附加的沖洗水Equipmentcanbeusedforfurtherprocessingwithoutadditionalsteps不需要附加步驟，設(shè)備用于進(jìn)一步的過程Resultscaneasilybeusedforcarryovercalculations結(jié)果易用于遺留計算Representswhatisleftonsurfacesafterthecompletionofthecleaningprocess代表了完成清潔過程后什么留在了表面Morelikelytoresultinanacceptableresultifdonecorrectly如果正確完成，更有可能產(chǎn)生可接受的結(jié)果Disadvantages劣勢Samplerepresentsaworst-casecarryovertothenextbatch(butcandemonstraterobustnessofthecleaningprocess)樣品代表了對下一批次的最差情況的遺留(但是能夠顯示清潔過程的穩(wěn)定性)Utilizesanadditionalstep使用附加步驟Requiresadditionalamountsofrinsewater要求附加的沖洗水量Contaminationispossibleduetothemethodofwateraddition由于補充水Assumptionsneedtobemadeaboutsamplingforcarryovercalculations制定假設(shè)關(guān)于遺留計算的取樣的方法引入污染Onlinetestingnotpractical在線檢測不實際Aspecialcaseofrinsesamplingisthesamplingofsmallparts.Thosepartsmaybesampledbyswabbing,buttherearetwooptionsforrinsesampling.Onetypeofrinsesamplingisextractionofthesmallparts.Inanextractionprocedure,theextractionsolution(typicallywaterforbiotechnologyresidues)isplacedinacleanvessel.Thesmallpartisthenplacedintheextractionsolutionandagitatedorsonicatedforafixedtime.Thesamplingsolutionisthenanalyzedforpotentialresidues.Asecondtypeofrinsesamplingforsmallpartsistypicallyusedforitemswithanorifice,suchasfillingneedles.Inthisprocedure,afixedvolumeofsamplingsolution(again,typicallywater)ispassedthroughthelumenandcollectedinacleancollectionvessel.Thesamplingsolutionisagitatedforuniformity,andthenanalyzedforthepotentialresidues.Becausethesurfaceareaandsamplingvolumearepreciselyknown,limitscanbeaccuratelycalculatedforsuchsituations.沖洗水取樣的一個特殊情況就是小部件取樣。沖洗取樣的一種方法就是小部件提取。提取過程中，提取液(生物技術(shù)殘留的典型水)被放置在清潔的容器中。之后小部件被放置在提取液中并且在固定的時間進(jìn)行搖動或者超聲處理。之后對樣品溶液進(jìn)行分析可能存在的殘留。對于小部件沖洗取樣的第二種方法典型的用于具有孔口的部件，比如灌裝針頭。在這個程序中，定量的樣品溶液(再次，代表性的水)通過內(nèi)腔并在潔凈的收取容器中收集。勻速搖動樣品溶液，之后對可能存在的殘留進(jìn)行分析。因為表面區(qū)域和取樣量是精確已知的，對于這種情況，能夠精確計算限度。SwabSampling擦拭取樣Swabsamplinginvolveswipingasurfacewithafibrousmaterial(mostcommonly).Duringthewipingprocedure,theresidueonthesurfacemaybetransferredtothefibrousmaterial.Thefibrousmaterialisthenplacedinasolventtotransfertheresiduetothesolvent.Thesolventisthenanalyzedfortheresiduebyanacceptableanalyticaltechnique.Themostcommonfibrousmaterialissomekindoftextile(knitted,wovenornonwoven)attachedtoaplastichandle.擦拭取樣涉及使用纖維材料擦拭表面(最一般的)。擦拭過程中，表面的殘留被轉(zhuǎn)移至纖維材料上。隨后纖維材料被放置在溶劑中，將殘留轉(zhuǎn)移至溶劑中。之后采用可接受的分析技術(shù)對溶劑進(jìn)行殘留分析。最常見的纖維材料是帶有塑料手柄的一些紡織物(針織的，梭織的或者無紡布的)。Inmostcasestheswabsarewettedwithasolventpriortosamplingthesurface.ForTOCandconductivity,thesolventisalmostalwayswater.Forsamplingthesamesite,companiesmaychoosetosamplethesamesurfaceareawithmultipleswabsinordertoprovideahigherpercentrecoveryofresiduefromthesurface.Insuchcases,theadditionalswab(s)utilizedmaybeeitherdryswab(s)orswab(s)wettedwiththesamesolvent.大部分情況下，在對表面取樣前棉簽是濕的并且?guī)в腥軇?。對于TOC和電導(dǎo)的方法，溶劑大部分情況下總是水。對于同樣位置的取樣，公司可能選擇取樣同樣的表面積進(jìn)行多次擦拭，以提供表面更高的殘留回收率。在這些情況下，使用的附加擦拭可以是干擦或者具有同樣溶劑的濕擦。ComparisonofSwabandRinseSampling擦拭和沖洗取樣的比較Bothswabsamplingandrinsesamplingarelistedasacceptablesamplingtechniquesinmostregulatorydocuments.(8,9,11)Bothmethodshavetheiradvantagesanddisadvantages,asshowninTable5.1.4在大部分的法規(guī)文件中，作為可接受的取樣技術(shù)，擦拭取樣和沖洗水取樣都被列出。(8.9.11)。兩種方法都具有其各自的優(yōu)勢和劣勢，見表格5.1.4.Table5.1.4ComparisonofSwabSamplingandRinseSampling表格5.1.4擦拭取樣和沖洗水取樣的比較Swabsampling擦拭取樣Rinsesampling沖洗水取樣Advantages優(yōu)勢Enablestheanalysisofresiduesfoundonthespecificsurfaces.Includestherecoveryofproteinsthataredenaturedand/oradheredtothesurface.基于特定的表面進(jìn)行殘留的分析。包括變性和/或粘附于表面的蛋白質(zhì)的回收。Allowsforsamplingofareasthataremoredifficulttoclean(i.e.,worstcases).對于難以清潔的區(qū)域進(jìn)行取樣(即，最差情況)Duringrinsing,theentireproduct-contactingsurfaceiswetted.Oneanalysisresultrepresentsthesumofallremovedresiduesfortheflowpath.沖洗過程中，產(chǎn)品接觸的全部表面都是濕的。一個分析結(jié)果代表了沖洗過程中所有移除殘留的總和。Thesamplingproceduredoesnotcontaminatetheequipment.Re-cleaningisnotrequiredaftersampling.取樣過程不會對設(shè)備產(chǎn)生污染。取樣后不需要再次清潔。Thismethodallowsforconclusionsonthecleanlinessofareasthatarenotaccessibleforswabbing.這種方法允許對不能進(jìn)行擦拭的區(qū)域的清潔度作出結(jié)論。Disadvantages劣勢Onlydiscretesamplingareascanbeanalyzed,andthesemustrepresenttheentireequipment;samplingmustincludeworst-caselocations.只有離散米樣區(qū)域能夠被分析，這些必須代表整體設(shè)備，取樣必須包括最差情況位置。Thesamplingitselfcanpotentiallycontaminatetheequipment.Re-cleaningmayberequiredaftersampling.取樣本身能夠?qū)υO(shè)備產(chǎn)生潛在污染。取樣后要求進(jìn)行再次清潔。Someareasarenotaccessibleforswabbing(e.g.,pipingsystems).一些區(qū)域是不能進(jìn)行擦拭的。(如管道系統(tǒng))Onlywatersolubleresiduescanbedetected.只有水溶性的殘留能夠被監(jiān)測。Thoseareasthatarehardtocleancannotbeidentified.難易清潔的區(qū)域不能被識別。Doesnotdealwithresiduesthatpreferentiallytransferfromonepartoftheequipmenttothenextproduct.不能夠處理從設(shè)備一部分優(yōu)先轉(zhuǎn)移至下一個產(chǎn)品的殘留。Maydiluteouttheresiduetobeundetectablebytheanalyticalmethod. 通過分析方法可能會稀釋殘留而不能檢測。Incaseswheretheequipmentsurfaceisdifficulttoaccessforswabbing(e.g.,piping),swabbingisnotanoption.Itshouldbeappropriatelyjustifiedthatthecleaningprocedureisconsideredeffectiveifswabtestingwillnotbeperformed.Thefollowingsituationswilljustifyadecisiontonotswabasurface:在設(shè)備表面難以進(jìn)行擦拭取樣的情況下(如，管道)，擦拭方法就不是一個選擇。如果擦拭檢測不能夠進(jìn)行，應(yīng)適當(dāng)說明清潔程序被認(rèn)為有效的。下列情況將說明不進(jìn)行表面擦拭：Equipmentnotaccessibleforswabbingisconstructedofthesamematerialsasequipmentthatallowsswabbing.不能夠進(jìn)行擦拭的設(shè)備與能夠進(jìn)行擦拭的設(shè)備具有相同的材質(zhì)。Difficulttoaccessequipmentsurfacesareexposedtothesameresiduesandconditionsasequipmentsurfacesthatallowswabbing.難以接觸的設(shè)備表面暴露在與能夠進(jìn)行表面擦拭的設(shè)備相同的殘留和條件下。Difficulttoaccessequipmentsurfacesarecleanedwiththesamecleaningprocedure(i.e.,thesamecleaningagentsandthesametemperature)asequipmentthatallowsswabbing.采用與能夠進(jìn)行擦拭的設(shè)備相同的清潔程序?qū)﹄y以接觸的設(shè)備表面進(jìn)行清潔。(即，同樣的清潔劑和同樣的溫度)Themechanicalforcesduringcleaninginpipingsystems(e.g.,turbulentflow)arehighercomparedtotankcleaningusingsprayballs.管道系統(tǒng)中清潔過程的機(jī)械力與使用噴淋球清潔的罐體相比更大。Incontrasttotanks,thepipingsystemiscompletelyfilledwithflowingliquidduringcleaning.與罐體相比，清潔過程中管道系統(tǒng)完全充滿了流動的液體。Rinsesamplingappropriatelyaddressestheissueofcross-contaminationfromthosesurfaces.沖洗取樣適當(dāng)?shù)慕鉀Q了表面交叉污染的問題。PlaceboSampling空白對照取樣Inbiotechnology,placebosamplinggenerallydoesnotincludeactualproductplacebo;instead,itincludesonlyWFIortheaqueousprocessingbufferwithoutanyproduct.Inthissamplingprocess,theequipmentiscleaned.Followingcleaning,amanufacturingprocessisperformed(totheextentfeasible)usingonlyWFIorbuffer.Followingprocessing,theWFIorbufferisevaluatedasanyothercleaningvalidationsample,typicallyforTOC(orTotalProtein),conductivity,bioburdenandendotoxin,asmeasuresofpossiblecontaminationofamanufacturedproductwiththoseresidues.Placeborunscanbedoneforbothbulkandformulation/fillmanufacturingtodemonstrateactualcarryovertotheprocessedmaterial.在生物工程中，空白對照取樣通常不包括實際產(chǎn)品的空白對照，相反的，只包括注射用水或者沒有任何產(chǎn)品的水性處理緩沖液。在取樣過程中，設(shè)備被清潔。隨著清潔只用注射用水或者緩沖液進(jìn)行生產(chǎn)加工過程。隨著加工，注射用水或者緩沖液被認(rèn)為是任何其他的清潔確認(rèn)樣品，尤其是對TOC(或總蛋白)，電導(dǎo)率，生物載荷以及內(nèi)毒素，作為具有這些殘留的生產(chǎn)產(chǎn)品可能存在的污染的測量。對于散裝和制劑/灌裝生產(chǎn)進(jìn)行的空白對照顯示了真實的到處理過材料的遺留。SamplingforMicrobialandEndotoxinAnalysis微生物以及內(nèi)毒素分析取樣Samplingforbioburdeninvolvesrinsewatersampling,swabbingorcontactplates.Rinsewatersamplingforbioburdenmustinvolvetheuseofsterilesamplecontainers.Acarefulsamplingtechniqueisrequiredforanymicrobialmethodtoavoidexternalcontaminationofthesample.生物載荷的取樣涉及沖洗水取樣，擦拭或者接觸碟。生物載荷的沖洗水取樣必須包括無菌取樣容器的使用。一個細(xì)致的取樣技術(shù)對任何微生物方法都是被要求的，以避免取樣的外部污染。Samplingforendotoxinisalmostalwaysarinsewatersample.內(nèi)毒素取樣總是采用沖洗水取樣。Thereisnothinguniquetobiotechnologyabouttheuseofthesesamplingtechniquesformicrobialevaluation.關(guān)于微生物評估的這些取樣技術(shù)，對于生物技術(shù)不是唯一的。SamplingRecoveryStudies取樣回收研究Samplingrecoverystudiesaregenerallyrequiredtoadequatelydemonstratethataresidue,ifpresentonequipmentsurfaces,canbeadequatelymeasuredorquantifiedbythecombinationoftheanalyticalmethodandthesamplingprocedure.Thesestudiesprovideascientificbasisforutilizingthosesamplingandanalyticalmethodstomeasureresidues.Threetypesofsamplingrecoveriesarediscussedbelow:swabsamplingrecovery,rinsesamplingrecovery,and“visualexamination”recovery.取樣回收研究一般都是要求的，充分顯示如果存在在設(shè)備表面，殘留能夠通過分析方法的結(jié)合和取樣程序進(jìn)行監(jiān)測和定量。這些研究為使用檢測殘留的取樣和分析方法提供了科學(xué)依據(jù)。下面討論了三種取樣回收：擦拭取樣回收，沖洗水取樣回收以及“目檢”回收。Forswabandrinsesampling,recoverystudiesmaybeperformedaspartoftheanalyticalmethodvalidation,ortheymaybeperformedasseparatestudies,onceitisdeterminedthattheanalyticalmethodcanappropriatelymeasureresiduesinsolutions.Samplingrecoverystudiesarelaboratorystudiesinvolvingcouponsofsampledequipmentofdifferentmaterialsofconstruction(suchasstainlesssteel,glass,PTFEandsilicone)spikedwithresiduestobemeasured.對于擦拭和沖洗水取樣，回收研究作為分析方法驗證的一部分進(jìn)行，或者可以作為單獨的研究進(jìn)行，一旦決定分析方法能夠在溶液中檢測出殘留。取樣回收研究是實驗室研究，包括不同材質(zhì)的被取樣設(shè)備的取樣片(如，不銹鋼，玻璃，PTFE和硅膠)，帶有殘留以檢測。5.4.1GeneralConsiderations總貝。Recoverystudiesmaynotberequiredforcertainresidueswhichareknowntobereadilywatersolubleandareusedwellbelowthesolubilitylimit,suchassodiumhydroxideorphosphoricacidusedascleaningagents.回收研究對于已知是水溶的并且低于溶解限度的某些殘留是不需要的，如，作為清潔劑的氫氧化鈉或者磷酸。Inperformingrecoverystudiesforswabbingandrinsesampling,theamountofmaterialspikedontocouponsshouldrepresentanamountequaltowhatcouldbepresentattheresiduelimit,asthisrepresentsaworstcase.在進(jìn)行擦拭和沖洗水取樣回收研究中，帶有材質(zhì)的取樣片的數(shù)量應(yīng)代表等同于代表殘留限度的數(shù)量，這也代表了最差條件。Theresiduespikedshouldbethesameresiduepresentattheendofthecleaningprocess.Forbiotechnologyproteinactives,thisisactuallydegradedproteinfragment.However,itiscommonpracticetospikethenativeproteinactive,asthisissimplerandrepresentsaworstcase.Forbulkbiotechnologymanufacturing,somemanufacturersonlyperformrecoverywiththebulkactive,whereasotherswillalsoutilizeanearlystageharvestproducttorepresentearlystageresidues.標(biāo)記的殘留應(yīng)是與警戒過程結(jié)束時出現(xiàn)的一樣的殘留。對于生物技術(shù)蛋白活性物，實際是降解的蛋白質(zhì)碎片。但是慣例做法是標(biāo)記主體蛋白活性物質(zhì)，因為這樣更簡單并且代表了最差條件。對于大部分生物技術(shù)生產(chǎn)，一些生產(chǎn)生只對原料藥進(jìn)行回收，其他的將使用早期階段的產(chǎn)品代表早期階段的殘留。Recoveryvaluesshouldbeestablishedforallsurfacessampled.Forswabandrinsesampling,thismaybeaccomplishedbyperformingrecoverystudiesonallsurfaces.Analternativeistoperformoneresiduestudyonasurface,whichthroughdocumentedevidence,isequivalent(intermsofpercentrecovery)toothersurfacesforwhichaformalrecoverystudyisnotperformed.Thisisessentiallyagroupingorfamilyapproachforrecoverystudies.Equivalenceforestablishingthegrouporfamilymaybeestablishedbasedonpublishedstudiesorin-housedata.Anotherapproachusedbysomecompaniesistoexcludeformalrecoverystudiesforsampledsurfacesconstitutinglessthanasmallpercentage(suchas1%or2%)ofthetotalequipmentsurfacearea;insuchcases,therecoveryvalueusedforthatexcludedsurfaceisthelowestrecoveryofanyothersurfacetypeforwhichaformalsamplingrecoverystudywasperformed,ortheminimumacceptablerecoverypercentagerequiredbythecompany’sprocedures.對于所有取樣的表面都應(yīng)建立回收值。對于擦拭和沖洗水取樣，在進(jìn)行所有表面的回收研究時完成。可替換的選擇就是在表面上進(jìn)行一個殘留研究，通過記錄的證據(jù)，等同于(術(shù)語是回收率)其他沒有進(jìn)行正式回收研究的表面。對于回收研究，這是本質(zhì)的分組或者家族方法。建立分組或家族等值是基于發(fā)布的研究或內(nèi)部的數(shù)據(jù)。一些公司使用的另一個方法是排除整個設(shè)備表面少于小的百分比(如1%或2%)取樣表面組成的正式回收研究，在這樣的情況下，用于排除表面的回收值是任何其他表面種類的最低回收，為此進(jìn)行正式的取樣回收研究或者根據(jù)公司程序要求最低可接受回收百分比。Swabrecoverystudiesaretypicallyperformedonanominalcouponsquaresurfaceareaofeither25cm2or100cm2.Insamplingmanufacturingequipmentforaprotocol,itisnotalwayspossibletoswaba10cmX10cmarea(itmightbenecessarytoswaba5cmX20cmarea).Furthermore,itmightnotbepracticaltoswabexactly100cm2(anareaof60cm2or128cm2mayberequiredbecauseofthespecificequipmentgeometry).Insuchcases,therecoverypercentagebasedonsampling10cmX10cmmaybeappliedtoeachofthosecases.Ifsuchanapproachisused,arangeofacceptablesurfacearea(suchas25%to150%ofthenominalsampledarea)shouldbeestablished.However,ifthesampledareaforequipmentsurfacesinaprotocolvariesfromthenominalvalue,theresiduelimitforthatsampleshouldbeadjustedbasedontheactualsurfaceareaswabbed.擦拭回收研究被進(jìn)行是在平方面積為25cm2或者100cm2。方案中取樣生產(chǎn)設(shè)備中，擦拭10cmx10cm的區(qū)域并不總是可能的(必要時擦拭5cmx20cm的區(qū)域)。此外，精確的擦拭100cm2實際上是不可能的(可能要求60cm2或者128cm2，由于設(shè)備特定幾何結(jié)構(gòu))。在這種情況下，針對這些情況基于取樣10cmx10cm應(yīng)用回收百分比。如果采用這種方法，可接受表面積范圍(如，實際取樣區(qū)域的25%到150%)應(yīng)該建立。但是，如果方案中設(shè)備表面的取樣區(qū)域與實際有區(qū)別，樣品的殘留限度應(yīng)基于實際的擦拭面積進(jìn)行調(diào)整。5.4.2SwabRecovery擦拭回收Forswabrecoverystudies,couponsarespikedwithsolutionsofthetargetresidue,allowedtodry,andsampledwiththeswabbingproceduretobeutilizedinthecleaningvalidationprotocol.Theswabisdesorbedinasuitablesolvent,andtheamountofresidueismeasuredinthatsolventsample.Theamountrecoverediscomparedtotheamountspikedonthecoupon,andtheresultisexpressedaspercentrecovery.Becauseswabbingisamanualprocedure,typicallyeachpersonperformingarecoverystudyperformsthreereplicates.Itispreferabletohaveatleasttwopersonsperformswabbingrecoverystudiesforeachcombinationofresidueandsurfacetype.Therecoverypercentageestablishedbythestudymaybedefinedindifferentways,buttypicallyisdefinedasthelowestaveragerecoveryofanyoneanalyst.Anacceptableswabrecoverydependsonhowthatswabrecoveryisbeingused.Iftherecoveryisperformedtoqualifythesamplingmethodwithoutcorrectionofeitheralimitorananalyticalresult,arecoveryof70%ormoreistypicallyrequired.Iftherecoverypercentageisusedtocorrectaresiduelimitorananalyticalresult,arecoveryof50%ormoreistypicallyrequired.Anupperlimitforpercentrecoveryshouldbeestablished.對于擦拭回收研究，取樣片使用目標(biāo)殘留溶劑標(biāo)記，干燥，在清潔驗證方案中使用擦拭程序進(jìn)行取樣。在適當(dāng)?shù)娜軇┲胁潦帽唤馕?，在實際樣品中測量殘留的數(shù)量。回收量與取樣片上標(biāo)記的數(shù)量進(jìn)行比較，以回收百分比的形式表示結(jié)果。因為擦拭是手工程序，每個人重復(fù)進(jìn)行三次回收研究。最好是至少兩個人對每個殘留結(jié)合和表面種類進(jìn)行擦拭回收研究。研究所建立的回收百分比可以以不同的方式定義，但是代表性的被定義為任何一個分析者的最低平均回收率。一個可接受的擦拭回收率取決于如何使用擦拭回收率。如果進(jìn)行的回收率以證明取樣方法不具有限度或者分析結(jié)果的修正，要求至少70%的回收率。如果回收百分比被用于修正殘留限度或分析結(jié)果，要求至少50%的回收率。應(yīng)建立回收百分比的上限。Ataminimum,recoveryvaluesaregenerallyperformedattheresiduelimitonthesurface(inpg/cm2,forexample).Whileitispossibletoperformrecoveriesatdifferentspikedlevels,ingeneral,thereislittlevaluetosuchadditionalspikedlevelsbecauseofthevariabilityofthesamplingprocedure.Itispreferabletoperformadditionalreplicatesattheone-residuelimitratherthanstudiesatadditionallevels.Acceptablevariationforrecoveryresultsatonespikedlevelistypicallyontheorderof15-30%RSD.最低限度，回收值通常是表面的殘留限度(如，pg/cm2)。在不同標(biāo)記水平進(jìn)行回收是可能的，通常，對于這種附加標(biāo)記水平幾乎沒有值，由于取樣程序的可變性。最好在一個殘留限度進(jìn)行附加的重復(fù)而不是在附加水平上進(jìn)行重復(fù)研究。在一個標(biāo)記水平的回收結(jié)果的可接受變化是近似于15-30%RSD。RinseRecovery沖洗回收Rinserecoverystudiesaddressthevalidityofrinsesamplingforthatresidue.Theydemonstratethatiftheresiduewereonasurface,thatresiduewouldbeeffectivelyremovedandcouldbeanalyzedintherinsesolution.RinserecoverystudiesaddresstheU.S.FDA’s“dirtypot"and“baby/bathwater”analogies.(8)Rinserecoverystudies,likeswabrecoverystudies,areperformedoncouponsthathavebeenspikedwithsolutionsofthetargetresidueandthenallowedtodry.Forswabrecoveries,itisnecessarytoperformtheexactswabbingproceduretobeusedinthecleaningvalidationprotocol.Forrinsesampling,incontrast,theexactrinsingprocedurecannotbeduplicatedinthelaboratory.However,itispossibletosimulatetherinsingprocedureinthelaboratory.Wherepossible,theconditionsofthesimulatedrinseshouldbethesameastheequipmentrinsingsituation.Thisincludestheselectionofrinsingsolvent(typicallywater),aswellasthetemperatureoftherinsingsolvent.Inothercases,therinsingconditionsshouldbeselectedasthesameorworstcaseascomparedtotheequipmentrinsingsituation.Forexample,theratioofsolventtosampledsurfaceareashouldbethesameorlowerintherecoverystudycomparedtotheequipment-rinsingsituation。沖洗回收研究解決了殘留沖洗取樣的正確性。說明了如果表面有殘留，殘留將被有效移除并在沖洗液中進(jìn)行分析。沖洗回收研究解決了美國“臟罐”和“嬰兒/浴缸水”的類比(8)。沖洗回收研究像擦拭回收研究一樣，在標(biāo)有目標(biāo)殘留溶液的取樣片上進(jìn)行并允許干燥。對于擦拭取樣，在清潔驗證方案中使用準(zhǔn)確的擦拭程序是必要的。對于沖洗取樣，相反的，精確地沖洗程序在實驗室中是不能夠被復(fù)制的。但是，在實驗室中模擬沖洗程序是可以實現(xiàn)的。如果可能，模擬沖洗的條件應(yīng)該與設(shè)備沖洗的情況相同。這包括沖洗溶劑的選擇(典型的是水)，以及沖洗溶劑的溫度。在其他的情況下，應(yīng)選擇與設(shè)備沖洗條件相同或者最差情況的沖洗條件。比如，取樣表面的溶劑比在回收研究中應(yīng)與設(shè)備沖洗條件相同或者低于設(shè)備沖洗條件。Onemethodofsimulatingtherinseprocessistosuspendaspikedcouponaboveacleancollectionvesselandcascadetherinsesolutionacrossthesurfaceintothecollectionvessel.Anothermethodistospikethebottomofabeakeroftheappropriatematerialofconstruction,allowtheresiduetodry,addrinsesolutiontothebeakerandapplygentleagitationforatimewhichapproximatesthetimeofthefinalrinse.Therinsesolutioniseitherpipettedordecantedfromthebeakerandanalyzed.Athirdoption,usedincaseswhereabeakerofsuitablematerialofconstructionisnotavailable,istoplaceaspikedcouponinthebottomofabeakerandperformasimulatedrinse,asinthesecondsituation.模擬沖洗過程的一種方法是在清潔的收集容器之上懸掛標(biāo)記的取樣片并傾倒沖洗溶液到表面至收集容器中。另一種方法是標(biāo)記適當(dāng)材質(zhì)的燒杯底部，允許殘留干燥，加入沖洗溶液至燒杯，溫和攪拌一段時間，近似于最終沖洗的時間。沖洗溶液從燒杯中由移液管吸出或者倒出并進(jìn)行分析。第三種選擇，這種情況下使用材質(zhì)的燒杯是不合適的，在燒杯底部放入標(biāo)記的取樣片，模擬沖洗，后續(xù)如第二種方法。Sincerinsesamplingisnotsignificantlyoperatordependent,threereplicatesbyoneoperatorareadequatetodeterminepercentrecovery.Acceptablepercentrecoveriesaretypicallyestablishedatthesamelevelsandconditionsasforswabrecoverystudies.因為沖洗取樣不典型的依賴于操作者，一個操作人員重復(fù)三次以確定回收百分比是充足的?？山邮艿幕厥瞻俜直冉⒃诓潦没厥昭芯客瑯拥乃胶蜅l件下。5.4.4"Recovery”inVisualInspection目檢中的“回收”Thisprocessisactuallythedeterminationofaquantitative“visualdetectionlimit”wherevisualexaminationisthesolesampling/analyticalmethodand“visuallyclean”isusedasthesoleacceptance