OMCL儀器確認(rèn)-附件HPLC確認(rèn)-中英文

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QUALITYMANAGEMENTDOCUMENTPA/PH/OMCL(11)04R6QUALIFICATIONOFEQUIPMENT

ANNEX1:QUALIFICATIONOFLIQUIDCHROMATOGRAPHY

EQUIPMENTFulldocumenttitleandreferenceQualificationofEquipmentAnnex1:QualificationofLiquidChromatographyEquipmentPA/PH/OMCL(11)04R6DocumenttypeGuidelineLegislativebasisCouncilDirective2001/83/ECand2001/82/EC,asamended;ThepresentdocumentwasalsoacceptedbyEAasrecommendationdocumenttobeusedinthecontextofQualityManagementSystemauditsofOMCLsDateoffirstadoptionMay2005DateoforiginalentryintoforceJune2005DateofentryintoforceofreviseddocumentAugust2018Previoustitles/otherreferences/lastvalidversionThisdocumentreplacesPA/PH/OMCL(11)04CORRCustodianOrganisationThepresentdocumentwaselaboratedbytheOMCLNetwork/EDQMoftheCouncilofEuropeConcernedNetworkGEONPA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentANNEX1OFTHEOMCLNETWORKGUIDELINE

“QUALIFICATIONOFEQUIPMENT”QUALIFICATIONOFLIQUIDCHROMATOGRAPHYEQUIPMENTNote:Mandatoryrequirementsinthisguidelineanditsannexesaredefinedusingtheterms?shall?or?must?.Theuseof?should?mcatesarecommendation.Forthesepartsofthetextotherappropriatelyjustifiedapproachesareacceptable.Theterm?can?indicatesapossibilityoranexamplewithnon-bindingcharacter.Introduction簡介ThepresentdocumentisthefirstAnnexofthecoredocument“QualificationofEquipment”,anditshouldbeusedincombinationwithitwhenplanning,performinganddocumentingtheLiquidChromatography(LC)equipmentqualificationprocess.本文件是《設(shè)備確認(rèn)》核心文件的第一個附件，在液相色譜(LC)設(shè)備確認(rèn)過程的規(guī)劃、實(shí)施和文件編制中應(yīng)與本文件結(jié)合使用。Thecoredocument“QualificationofEquipment”containsthegeneralintroductionandtheLevelIandIIofqualification,commontoalltypesofinstruments.ThepresentAnnex1containsageneralintroductionandrequirementsforLC(HPLCandUHPLC)instruments.核心文件“設(shè)備確認(rèn)”包括一般介紹和所有類型的儀器共有的一級和二級確認(rèn)。本附件1包含了LC(HPLC和UHPLC)儀器的一般介紹和要求。LevelIIIandIVqualificationsmustbecarriedoutbeinganISO17025requirement.III級和IV級確認(rèn)必須符合ISO17025的要求。Requirementsand(ifapplicable)correspondingtypicalacceptancelimitsgiveninboldshouldbeapplied;howeverotherappropriatelyjustifiedapproachesareacceptable.要求和(如適用)相應(yīng)的以粗體顯示的典型接受限度應(yīng)適用；然而，其他合理的方法是可以接受的。ExemplaryproceduresprovidedinAnnexeshavenon-bindingcharacter.Theycanbehelpfultocarryouttherequiredqualification.Nevertheless,itislefttotheprofessionaljudgementandbackgroundexperienceofeachOMCLtodecideonthemostrelevantprocedurestobeundertakeninordertogiveevidencethattheirLCsystemsareworkingproperlyandaresuitablefortheirintendeduse.附件中提供的示范程序具有非約束性。他們可以幫助執(zhí)行所需的確認(rèn)。然而，每一個綜合管理公司的專業(yè)判斷和背景經(jīng)驗(yàn)，決定采取最相關(guān)的程序，以證明其LC系統(tǒng)工作正常，適合其預(yù)期用途。Moreover,combinedtestprocedurescanbeappliedtocarryoutLevelIIIandIVqualificationstocheckseveralparameterssimultaneously(e.g.“overall”systemperformancetestforpeakareaprecision,retentiontimeprecision,gradientreproducibility,etc.)止匕外，可以應(yīng)用組合測試程序進(jìn)行III級和IV級確認(rèn)認(rèn)證，同時檢查多個參數(shù)(例如，“整體”系統(tǒng)性能測試的峰面積精度、保留時間精度、梯度再現(xiàn)性等)。p.2/25

PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentLevelIII.Periodicandmotivatedinstrumentchecks定期和積極的儀器檢查RecommendationsforLCinstrumentsandrelatedtypicalacceptancelimitsLC儀器的建議和相關(guān)的典型接受限度InstrumentmoduleParametertobecheckedTypicalAcceptancelimitsSolventdeliverysystem溶劑輸送系統(tǒng)Flowrateaccuracy(HPLC)流速準(zhǔn)確度(HPLC)±5.0%Flowrateaccuracy(UHPLC)流速準(zhǔn)確度(UHPLC)±3.0%Flowrateprecision(HPLCandUHPLC)流速精度(HPLC和UHPLC)RSD<0.5%Gradientcompositionaccuracy梯度成分準(zhǔn)確度±2.0%Gradientripple梯度波動±0.2%Injector注射器Volumeprecision(HPLCandUHPLC)體積精度(HPLC和UHPLC)RSD<1.0%Carry-over殘留<0.2%Autosampler自動進(jìn)樣器Thermostatingaccuracy恒溫準(zhǔn)確度±3°COvenorcoolingdevice(column)爐或冷卻裝置(柱)Thermostatingaccuracy恒溫準(zhǔn)確度±2°CThermostatingstability恒溫穩(wěn)定性<1°CMultiwavelengthdetector多波長檢測器Linearity線性r2>0,9990Wavelengthaccuracy波長準(zhǔn)確度±2nmDrift漂移AnnexIRefractiveIndexdetector示差折光檢測器Signal/Noiseratio信噪比>10Driftovertime時間漂移±0.1mV/minLinearity線性r2>0,9950Fluorescencedetector. Wavelengthaccuracy. (excitationandemission)Signal/Noiseratio3nm>400Electrochemicaldetectors:p.3/25

PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipment-Amperometricdetection**Drift(cellcurrent)Noise<8pA/h<2pA"IntegratedAmperometricdetection***LinearityDriftNoiser2>0,999<1250pC/20minMW20readouts<160pC"Conductivitydetection***LinearityDriftNoiser2>0,999<2nS<20nS/h"CoulometricdetectionAnnexIAnnexICircularDichroismdetectorLinearitySignal/NoiseratioDriftovertimeSpectracomparisonr2 0.9950>1,0<0.1mdeg/h±4nmChargedAerosoldetectorBaselinenoiseLargestRandomspikeBaselinedriftRepeatabilitySignal/NoiseratioSignalcalibration<0.04pA<0.2pA<0,04pA/minRSD10%>10r20.9990EvaporativeLightScatteringdetector**NoiseBaselinedriftRepeatability<2mV<2,0mV/hRSD3.0%p.4/25

PA/PH/OMCL（11）04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentLevelIV.In-useinstrumentchecks在用儀器檢查RecommendationsforLCinstrumentsandrelatedtypicalacceptancelimitsLC儀器的建議和相關(guān)的典型接受限度Parametertobechecked待校驗(yàn)參數(shù)Typicalacceptancelimits典型的驗(yàn)收范圍Systemsuitability系統(tǒng)適用性AccordingtoPh.Eur.orMAHdossierorvalidatedin-housemethod根據(jù)EP或MAH檔案或內(nèi)部驗(yàn)證方法Peakareaprecision（Assay,applicabletothemainpeakoftheanalytewhennotsaturated）峰面積精度（測定，適用于待測物未飽和時的主峰）Peakareaprecision（Relatedsubstances）峰面積精度（有關(guān)物質(zhì)）RSD<1.5%（min.5injectionsoftestorreferencesol.至少5針供試品或?qū)φ杖芤海╱nlessotherwiseprescribedinthesystemsuitabilityofthemethod,e.g.specificrequirementsfromPh.Eur.Chapter2.2.46,Ph.Eur.Monographs,orMAHdossiers除非該方法的系統(tǒng)適用性另有規(guī)定，例如，來自Ph.Eur的具體要求。第2.2.46章，專著，或MAH檔案）RSD<5.0%（minimum3injectionofthedilutedsolutionorreferencesolutionusedforquantification至少3針用于定量的稀釋溶液或?qū)φ杖芤海╱nlessotherwiseprescribedinthesystemsuitabilityofthemethod,e.g.specificrequirementsfromPh.Eur.Chapter2.2.46,Ph.Eur.monographsorMAHdossiers除非該方法的系統(tǒng)適用性另有規(guī)定，例如，來自Ph.Eur的具體要求。第2.2.46章，專著，或MAH檔案）Retentiontimeprecision（applicabletothemainpeakofthestandardsolutionwhennotsaturated）保留時間精度（適用于標(biāo)準(zhǔn)溶液未飽和時的主峰）RSD<2.0%（min.5injectionsoftestorreferencesol.至少5針供試品或?qū)φ杖芤海〤arry-over（bycomparingconsecutiveinjectionsofastandardsolutionofthesubstancebeingquantifiedandablankinjection）殘留（通過比較連續(xù)進(jìn)樣被定量物質(zhì)的標(biāo)準(zhǔn)溶液和空白進(jìn)樣）<0.2%（Assay含量）Belowdisregardlimit下面不限制（Relatedsubstances有關(guān)物質(zhì)）Signal/Noiseratio（tobeappliedforrelatedsubstancestestonly）信噪比（只適用于有關(guān)物質(zhì)測試）AccordingtoPh.Eur.2.2.46參照Ph.Eur.2.2.46.（unlessotherwiseprescribedinPh.Eur.monographsorMAHdossier除非EP另有規(guī)定。專著或MAH檔案）p.5/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentANNEXILevelIII.Periodicandmotivatedinstrumentchecks第三級定期性的主動儀器檢查ThisAnnexcontainspracticalexamplesofproceduresandtheirtypicalacceptancelimitsforseveralparameterstoperformtheLevelIIIqualificationofdifferentmodulesofHPLC/UHPLCinstruments.本附件包含了實(shí)際的程序示例和幾個參數(shù)的典型接受限度，以執(zhí)行HPLC/UHPLC儀器的不同模塊的III級認(rèn)證。Incasemorethanonemethodisdescribedfortestingoneparameter,method1ormethod2ormethod3canbeused.如果描述了用于測試一個參數(shù)的多個方法，則可以使用方法1或方法2或方法3。Forqualificationprocedure,wheneveracolumnisdescribed,itcanbeconsideredtouseadedicatedcolumn.對于確認(rèn)過程，無論何時描述一個列，都可以認(rèn)為使用了一個專用列。SOLVENTDELIVERYSYSTEM溶劑輸送系統(tǒng)ThefollowingtestsareproposedfortheperiodicandmotivatedcheckoftheHPLCsolventdeliverysystem:flow1ateandgradienttest.建議對高效液相色譜溶劑傳遞系統(tǒng)進(jìn)行定期和有動機(jī)的檢查:流速和梯度測試。FLOWRATEACCURACYandPRECISION流速準(zhǔn)確度和精密度Method1方法1Materials材料?Beaker(capacityof5-10mL)orweighingvessel燒杯(容量5-10毫升)或稱量器Calibratedthermometer校準(zhǔn)的溫度計Calibratedchronometer校準(zhǔn)的時鐘Calibratedanalyticalbalance校準(zhǔn)的分析天平Settings設(shè)置?Mobilephase:degassedwaterforchromatographyRCapillary流動相：脫氣的水，用于毛細(xì)管色譜e.g.2000x0.12mmcapillaryforbackpressureFlowrate:例如：2000x0.12mm毛細(xì)管回流壓力流量：HPLCsystem:0.5mL/minand5.0mL/minormaximumflowrateusedHPLC系統(tǒng)：0.5mL/min和5.0mL/min或使用的最大流速UHPLCsystem:0.2mL/minand2.0mL/minormaximumflowrateusedUHPLC系統(tǒng)：0.2mL/min和2.0mL/min或使用的最大流速Ifhigh-pressuremixingsystemsareinstalled,thistesthastobedoneoneachsolventchannel.如果安裝了高壓混合系統(tǒng)，則必須對每個溶劑通道進(jìn)行測試。p.6/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentProcedure程序：Settheflowrateatadesiredlevel(atleasttestoneflowrate,theloweroperationallevelinuse)andleavewatertoflowintothebeakerorweighingvessel,previouslyweighed(empty).Afteradefinedtime(e.g.5.0min)weighthebeakeragaincontainingthedeliveredwater.Measurethetemperatureofthewaterused.Repeattheprocedureatleast3times.Calculatethevolumedeliveredtothebeakerandtheflowrate:將流量設(shè)置在所需的水平(至少測試一個流量，使用中較低的操作水平)，讓水流入燒杯或稱重容器，之前稱重(空)。在規(guī)定的時間(如5.0分鐘)后，再次稱量盛有送出的水的燒杯。測量用水的溫度。重復(fù)以上步驟至少3次。計算送至燒杯的體積和流量：V=m/Qm weightofwaterdeliveredinthemeasuredtime實(shí)測時間內(nèi)送水重量[g]q densityofwateratactualtemperaturefromtable實(shí)際溫度下的水的密度[g/mL]V volumedelivered輸送量[mL]f=V/tf measuredflowrate實(shí)測流速[mL/min]t elapsedtime運(yùn)行時間[min]V volumedelivered輸送量[mL]D deviation偏差[%]F nominalflowrate名義流速[mL/min]f measuredflowrate實(shí)測流速[mL/min]Method2方法2Usecalibratedflowmeterfordeterminationofflowrate使用校準(zhǔn)過的流量計來測定流量Method3方法3Materials材料?Usecalibratedvolumetricglasswaree.g.flaskof5.0or10.0mLclassA使用校正后的容量玻璃器皿，例如5.0或10.0ml的A級量瓶Calibratedchronometer校正后的時鐘Settings設(shè)置Mobilephase:degassedwaterforchromatographyRCapillary流動相：脫氣的水，用于毛細(xì)管色譜e.g.2000x0.12mmcapillaryforbackpressureFlowrate:p.7/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipment例如：2000x0.12mm毛細(xì)管回流壓力流量：ForHPLCsystem:0.5mL/minand5.0mL/minormaximumflowrateusedHPLC系統(tǒng)：0.5mL/min和5.0mL/min或使用的最大流速ForUHPLCsystem:0.2mL/minand2.0mL/minormaximumflowrateusedUHPLC系統(tǒng)：0.2mL/min和2.0mL/min或使用的最大流速Ifhigh-pressuremixingsystemsareinstalled,thistesthastobedoneoneachsolventchannel.如果安裝了高壓混合系統(tǒng)，則必須對每個溶劑通道進(jìn)行測試。Procedure程序：Settheflowrateatanappropriatelevelandmeasurethetimeneededtofillthevolumetricflaskuptothemark.Recordthetimeneeded.將流速設(shè)置在適當(dāng)?shù)乃?，并測量將量瓶灌滿所需的時間。記錄所需的時間。,Vx60J二 tf measuredflowrate實(shí)測流速[mL/min]t elapsedtimetofilluptomark填充到標(biāo)記所用的時間[s]V volumeofthevolumetricflask量瓶的容積[mL]f—FD=100xf——FD deviation偏差[%]F nominalflowrate名義流速[mL/min]f measuredflowrate實(shí)測流速[mL/min]Limits限度：Accuracy準(zhǔn)確度：HPLC:D<5.0%UHPLC:D<3.0%Precision精密度(HPLCandUHPLC):Therelativestandarddeviation(RSD)shouldbe<0.5%.GRADIENTCOMPOSITIONACCURACYANDRIPPLE梯度成分準(zhǔn)確度和波動Settings設(shè)置Capillarye.g.2000x0.12mmforbackpressure毛細(xì)管，例如2000x0.12毫米的背壓Detection檢測波長：265nmMobilephase流動性A:waterforchromatographyR色譜用水Mobilephase流動相B:waterforchromatographyRcontaining0.5%acetone0.5%的丙酮水溶液Flowrate流速：1.0mL/minp.8/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentProcedure程序:Thetestiscarriedoutinthefollowingwaybyusingagradientprogramdependingonthenumberofsolventchannelsandtheconfiguration0fthesystem：根據(jù)溶劑通道的數(shù)量和系統(tǒng)的配置，使用梯度程序進(jìn)行測試，方法如下：A-BA-BandA-CA-C,A-BandB-DTableIII-ExampleforA-Bsystemconfiguration:A-B系統(tǒng)配置示例:time[min]%mobilephaseA(water)%mobilephaseB(water/acetonemixture)0.010000.1901010901010.1505020505020.1109030109030.1010040010040.11000Startthetestbypumpingwaterforatleast10mintoequilibratethesystem.開始測試時，泵水至少10分鐘以使系統(tǒng)達(dá)到平衡。Thezero%valueatthestartofthetestisthebaseline.Allstepsaremeasuredatthebeginningofthehorizontalpartofthelineeitherbysoftware(asabsorptionunits)ormanuallyonthepaperprintusingaruler.Theheightofthe100%water/acetonemixtureisusedasHinthefollowingcalculation.測試開始時的0%值是基線。所有的步驟都是通過軟件(作為吸收單元)或使用尺子在紙上打印的水平部分的開始處測量的。下面計算時用100%水/丙酮混合物的高度作為H。%h=iuo*Ap.9/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipment%H calculatedcomposition計算組分h heightofthemeasuredline測量線的高度H heightofthe100%water/acetonemixtureline(mobilephaseB)100%水/丙酮混合線高度（移動B相）d deviation偏差G nominalgradientcomposition[%water/acetonemixture=mobilephaseB]名義梯度成分[%水/丙酮混合物=流動相B]Limits限度：Accuracy準(zhǔn)確度：±2.0%Therippleofthegradientcompositionisthepercentageofnoiseofthe50%linefromthegradientprogram.梯度構(gòu)成的波動度是梯度程序中50%的線的噪聲百分比。用。%R ripple波動h50 heightofthe50%line50%線的高度N heightofthenoise,i.e.thedifferencebetweentheminimumandthemaximumofthe50%linemeasuredduring1minuteinthelinearregion噪聲的高度，即在線性區(qū)域內(nèi)1分鐘內(nèi)測量到的50%線的最大值與最小值的差值Limit限度：S0.2%INJECTOR注射器Volumeprecisionandcarry-overarethetestsproposedfortheperiodicandmotivatedcheckoftheLCinjector.體積精密度和殘留是針對LC注射器的定期和有動機(jī)的檢查提出的測試。VOLUMEPRECISIONANDCARRY-OVER體積精密度和殘留Method1方法1Solutions溶液。SolventA:waterforchromatographyR:methanolR(40:60V/V)溶液A：色譜用水:甲醇(40:60V/V)Referencesolution(a):dissolve15.0mgmethylparabenand15.0mgofpropylparabeninsolventAanddiluteto100.0mLwiththesamesolvent.參考溶液(a)：將15.0mg對羥基苯甲酸甲酯和15.0mg對羥基苯甲酸丙酯溶于溶劑A中，用相同的溶劑稀釋至100.0mL。p.10/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentReferencesolution(b):Dilute1.0mLofreferencesolution(a)to10.0mLwithsolventA.參考溶液(b):用溶劑a將1.0mL的參考溶液(a)稀釋至10.0mL。Settings設(shè)置?Column:Lichrospher100RP8,5pm,125x4mm,withoutpre-columnorequivalent色譜柱：Lichrospher100RP8,5pm,125x4mmMobilephase:waterforchromatographyR:methanolR(40:60V/V)流動相：色譜用水:甲醇(40:60V/V)Flowrate:1.0mL/min流速：1.0mL/minDetection:254nm檢測波長：254nmInjectionvolume進(jìn)樣體積：HPLC(“shortpathway”flow-cell短路徑流通池,forexample5or10mm):20pLUHPLC(“shortpathway”flow-cell短路徑流通池,forexample5or10mm):10pLHPLC/UPLC(“l(fā)ongpathway”flow-cell長路徑流通池,forexample60or85mm):2pLProcedure程序：Injectionscheme進(jìn)樣方案：1xsolventA(blankinjection1)6xreferencesolution(b)1xreferencesolution(a)1xsolventA(blankinjection2)1xreferencesolution(b)Limits限度：Repeatabilityofpeakareas:Therelativestandarddeviationofthepeakareasofmethylparabenandpropylparabeninthechromatogramsofthesixconsecutiveinjectionsofreferencesolution(b)shouldbe<1.0%峰面積重復(fù)性:連續(xù)六次注射參考溶液(b)色譜中對甲苯甲酸甲酯和對丙苯甲酸丙酯峰面積的相對標(biāo)準(zhǔn)偏差應(yīng)《1.0%Carry-over:Thepercentageofthepeakareacorrespondingtopropylparabenintheblankinjection2doesnotexceed2%ofthepeakareaofthepropylparabenpeakinthechromatogramobtainedwithreferencesolution(b)injectedaftertheblankinjection2,thiscorrespondsto0.2%殘留:空白注射2中對羥苯甲酸丙酯峰所對應(yīng)的峰面積百分比不超過空白注射2后用(b)對照液注入色譜圖中對羥苯甲酸丙酯峰所對應(yīng)的峰面積的2%，即為0.2%。Method2方法2Solutions溶液。Referencesolution(a):0.5mgcaffeine/mLwaterforchromatographyR參考溶液(a):0.5mg/mL咖啡因水溶液Referencesolution(b):50.0pgcaffeine/mLwaterforchromatographyR參考溶液(b)：50.0Pg/mL咖啡因水溶液p.11/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentSettings設(shè)置.Column色譜柱:RP-18,5pm,30-150x2.1-4.6mmMobilephase:waterfOrchromatographyR:acetonitrileR(85:15V/V)流動相：水：乙月青(85:15V/V)Oventemperature柱溫：40°CFlowrate流速：1.0mL/min(flowratemaybereducedaccordingtocolumndiameter)Detection檢測波長:273nmInjectionvolume進(jìn)樣體積：HPLC(“shortpathway”flow-cell,forexample5or10mm):20pLUHPLC(“shortpathway”flow-cell,forexample5or10mm):10pLHPLC/UPLC(“l(fā)ongpathway”flow-cell,forexample60or85mm):2pLProcedure程序：Injectionscheme進(jìn)樣方案：1xMobilephase(blankinjection1)6xReferencesolution(b)1xReferencesolution(a)1xMobilephase(blankinjection2)1xReferencesolution(b)Limits限度：Volumeprecision:Therelativestandarddeviationofthepeakareasofcaffeineinthechromatogramsofthesixconsecutiveinjectionsofreferencesolution(b)shouldbeW1.0%體積精密度:連續(xù)六次注射參考溶液(b)色譜中咖啡因峰面積的相對標(biāo)準(zhǔn)偏差應(yīng)勺.0%Carry-over:Thepercentageofthepeakareacorrespondingtocaffeineintheblankinjection2doesnotexceed2%thepeakareaofthecaffeinepeakinthechromatogramobtainedwithreferencesolution(b)injectedaftertheblankinjection2,thiscorrespondsto0.2%.殘留:空白注射劑2中咖啡因所對應(yīng)的峰面積百分比不超過空白注射劑2后用(b)對照液注射得到的色譜中咖啡因峰面積的2%,相當(dāng)于0.2%。AUTOSAMPLER自動進(jìn)樣器Thermostatingaccuracycanbetestedintheframeoftheperiodicandmotivatedcheckoftheautosampler.恒溫的準(zhǔn)確性可以在自動取樣器的定期主動檢查的框架內(nèi)進(jìn)行測試。THERMOSTATINGACCURACY恒溫準(zhǔn)確度Materials材料?Calibrated(electronic)thermometerwithasuitableprobe.用適當(dāng)?shù)奶结樞?zhǔn)(電子)溫度計。Procedure程序：Selectatemperaturealongtheoperationalorrequiredtemperaturerangeoftheequipment(e.g.thelowestpointcanbechecked,forexample+5°C,ortypicaltemperatureused).Waituntilthesystemisp.12/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentequilibrated.選擇一個溫度沿操作或所需的溫度范圍內(nèi)的設(shè)備(如最低點(diǎn)可以檢查，例如+5°C,或使用典型的溫度)。等到這個體系達(dá)到平衡。Bymeansofthecalibratedthermometer,measuretheactualtemperatureintheautosamplerandcompareittotheselectedtemperature.Asanalternativeprocedure,fillavialwithwater,waituntilequilibrationandmeasurethetemperatureofthewaterbyusinganappropriateprobe.Ifapplicable,repeatthemeasurefromseveralpointsoftheautosampler(mapping)followingtheinstructionsprovidedbythemanufacturer.用校準(zhǔn)過的溫度計測量自動取樣器的實(shí)際溫度，并與選定的溫度進(jìn)行比較。另一種方法是，將小瓶裝滿水，等水達(dá)到平衡后，用合適的探針測量水的溫度。如果適用，按照制造商提供的說明，從自動采樣器(映射)的幾個點(diǎn)重復(fù)測量。Limits限度：Theactualtemperatureshouldbewithin±3°Cwithrespecttheselectedtemperature.實(shí)際溫度應(yīng)在選擇溫度的±3℃以內(nèi)OVEN/COOLINGDEVICE柱溫箱/冷卻裝置Thermostatingaccuracyandstabilityaretheparameterstestedinthisexampleofperiodicandmotivatedcheckoftheoven/coolingdevice.恒溫的準(zhǔn)確性和穩(wěn)定性是在柱溫箱/冷卻裝置的定期和有動機(jī)的檢查中測試的參數(shù)。THERMOSTATINGACCURACYANDSTABILITY恒溫準(zhǔn)確度和穩(wěn)定性Materials材料?Calibratedthermometer.經(jīng)校準(zhǔn)的溫度計Procedure程序：Setthecolumnoventemperatureto40°C,waitabout30minutestoequilibratethesystem,putacalibratedthermometerintotheovenandtake6temperaturereadingsat4minutesintervals.將柱溫箱溫度設(shè)置為40℃，等待約30分鐘使系統(tǒng)達(dá)到平衡，將校準(zhǔn)后的溫度計放入柱溫箱，每隔4分鐘讀取6個溫度讀數(shù)。Ifapplicable,repeatthisproceduresettingtheoven/cooleratotheroperationaltemperaturesbasedontheworkingrangeoftemperatureoftheequipment.如適用，重復(fù)此程序，根據(jù)設(shè)備的工作溫度范圍，將柱溫箱/冷卻器設(shè)置為其他工作溫度。Theaccuracyiscalculatedasaverageof6temperaturereadings.精度計算為6個溫度讀數(shù)的平均值。Limits限度土2°CThestabilityisverifiedusingtheresultsobtainedintheaccuracytest.Thestabilityiscalculatedforeachtemperatureasthedifferencebetweenthehighestandthelowesttemperatureofthesixreadings.利用精度試驗(yàn)結(jié)果驗(yàn)證了該方法的穩(wěn)定性。穩(wěn)定性是根據(jù)六種讀數(shù)的最高和最低溫度之間的差值來計算的。Limits限度<1°Cp.13/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentMULTIWAVELENGTHDETECTOR多波長檢測器TheperiodicandmotivatedcheckoftheLCUV/visibleandDADdetectorscanbeperformedbytestingthelinearity,wavelengthaccuracyanddrift.通過測試線性度、波長精度和漂移，可以對lcUV/visible和dad檢測器進(jìn)行定期和有動機(jī)的檢查。LINEARITY線性Solutions溶液?Std.1:0.5pg/mLcaffeineStd.2:1.0pg/mLcaffeineStd.3:5.0pg/mLcaffeineStd.4:25.0pg/mLcaffeineStd.5:50.0pg/mLcaffeineStd.6:waterforchromatographyR(blank)水(空白)Forexample,prepareastocksolution(Std.0)byweighingabout10.0mgofcaffeineandfillupto20.0mLwithwaterforchromatographyR(dissolveusingsonicationormix).Dilutethissolution,forexamplefollowingthesu^cqucnt5cheme:例如，準(zhǔn)備一種原液(Std.0)，稱取大約10.0mg的咖啡因，用20毫升的水進(jìn)行色譜R(用超聲波或混合法溶解)。稀釋此溶液，如后續(xù)方案：Std.5:dilute10.0mLofstocksolutionto100.0mL將10.0mL原液稀釋至10somLStd.4:dilute5.0mLofstocksolutionto100.0mL將5.0mL原液稀釋至10somLStd.3:dilute1.0mLofstocksolutionto100.0mL將1.0mL原液稀釋至10somLStd.2:dilute10.0mLofStd.3to50.0mL將佃。mLStd,3稀釋至50.0mLStd.1:dilute10.0mLofStd.3to100.0mL將佃。mLStd,3稀釋至100.0mLUsewaterforchromatographyRtodilute.用色譜用水進(jìn)行稀釋。Settings設(shè)置?Column:suitablecolumnorcapillary(e.g.RP18)色譜柱：合適的色譜柱或毛細(xì)管(e.g.RP18)Mobilephase:acetonitrileR:waterforchromatographyR(15:85V/V)流動相：乙月青：水(15:85V/V)Oventemperature柱溫：40°CFlowrate流速：HPLC1.0mL/minUHPLC0.5mL/min(orappropriateflowratebasedoninstrumentspecifications/operationaluse)Detection檢測波長:273nmInjectionvolume進(jìn)樣體積：HPLC(“shortpathway”flow-cell,forexample5or10mm):20pLUHPLC(“shortpathway”flow-cell,forexample5or10mm):10pLp.14/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentHPLC/UPLC(“l(fā)ongpathway”flow-cell,forexample60or85mm):2^L(orappropriatevolumebasedoninstrumentspecifications/operationaluse)Procedure程序：Injectionscheme進(jìn)樣方案：2xblank1xStd.11xStd.21xStd.31xStd.41xStd.5Limits限度:r2>0.9990Remark:Asthistestemploysdifferenttestsolutionstobeinjected,itcoversalsothecheckofcorrectpositioning0fvialsintheautosampler.注:由于本試驗(yàn)采用不同的注入測試溶液，因此還包括在自動取樣器中檢查小瓶的正確位置。WAVELENGTHACCURACY波長準(zhǔn)確度Wavelengthaccuracy(andrelatedadjustment)canbecarriedoutbybuilt-intestprocedures,followinginstructionsoftheinstrumentmanual/manufacturer.Inallothercasesusetheproceduredescribedbelow.波長準(zhǔn)確度(和相關(guān)的調(diào)整)可以通過內(nèi)置的測試程序來執(zhí)行，遵循儀器手冊/制造商的說明。在所有其他情況下，使用下面描述的過程。Solutions溶液。CaffeineStd.5fromthelinearitytesting線性測試中的咖啡因std.5Settings設(shè)置?Usetheoperationalconditionsdescribedunderlinearitytestingexceptfordetection.除檢測器外，使用線性測試中描述的操作條件。Procedure程序：Injectthecaffeinesolutionandrecordthespectrumbyscanningfrom190to290nm(1nmincrementalsteps,wheneverpossible).Themaximaareat205nmand273nm,theminimumat245nm.注入咖啡因溶液，掃描190~290nm(可能的話，每增加1nm)，記錄光譜。吸收最大值為205nm，最大值為273nm，最小值為245nm。Limits限度：±2nmDRIFT漂移Typicallydriftverificationiscarriedoutbybuilt-intestprocedures.Followtheinstructionsoftheinstrumentmanual/manufacturcr.通常通過內(nèi)置的測試程序進(jìn)行漂移驗(yàn)證。按照儀器說明書/制造商的說明操作。Limits限度：accordingtomanufacturer.根據(jù)制造商。p.15/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentFLUORESCENCEDETECTORThefollowingthreeparametersareproposedfortheperformanceoftheperiodicandmotivatedcheckoftheLCfluorescencedetector:WAVELENGTHACCURACYFOREXCITATIONMethod:Rinseandfillthemeasuringcellwithde-ionizedwaterAdjusttheexcitationwavelengthto350nmandemissionwavelengthto397nm.Measuretheexcitationspectrum.Limitsforexcitationmaximum:350 3nmWAVELENGTHACCURACYFOREMISSIONMethod:Rinseandfillthemeasuringcellwithde-ionizedwaterAdjusttheexcitationwavelengthto350nmandemissionwavelengthto397nm.Measuretheemissionspectrum.Limitsforemissionmaximum:397 3nmSIGNALTONOISERATIOMethod:Rinseandfillthemeasuringcellwithde-ionizedwater.Set-uptheexcitationwavelengthat350nmandtheemissionwavelengthat397nm.Thesignalisacquiredoveratotalof23minutes.At20.30minutestheemissionwavelengthisswitchedto450nm.ThesignalismeasuredatthemaximumwavelengthofRamanband(i.e.397nmfor350nmexcitation)andthenoiseinaregion(450nm)wherenoRamansignalispresent.Signal-to-noiseiscalculateddividingtheheightoftheRamanbanddividedbythenoise.Limits:Signaltonoise>400ELECTROCHEMICALDETECTORThereareseveraltypesofelectrochemicaldetectorsavailablefromdifferentmanufacturers,basedonamperometry,coulometryorconductivitymethodofdetection.Amperometricdetectorp.16/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentDRIFTANDNOISESettings:Cellpotentialofadummycell:800mVRisetimefilter:0.1sRange:0.1nATemperature:30°CProcedure:Drift:Measuretheelectriccurrentandsubtract2.67nA(theoreticalvalue)Noise:Measurethenoiseoveraperiodof5minutesLimits:Drift(cellcurrent):<8pA/hNoiseofthesignal:<2pAor20mVIntegratedamperometricdetectorDRIFTANDNOISENoise:Readthesignaleveryminute.Settings:Pressure:>1000psiFlow:1.0mL/minRestrictioncapillary:PEEK,150cmx0.13mmColumn:CarboPacPA1GuardEluent:50mMNaOH,filteredthrough0.2^mNylon,degassedDetectorsettings:Waveform“AminoAcids”(pH,Ag,AgClReference)Runtime:20minLimits:Drift:<1250pC/20minNoise:MW20readout<^。pCLINEARITYSolutions:1,2,4,8,and10^MofThreonineinwaterforchromatographyRSettings:Pressure:>1500psiFlow:0.25mL/minRestrictioncapillary:PEEK,150cmx0.13mmColumn:CarboPacPA1Guard2mmEluent:50mMNaOH,filteredthrough0.2^mNylon,degassedp.17/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentDetectorsettings:Waveform“AminoAcids”(pH,Ag,AgClReference)Duration/runtime:3minInjectionvolume25^LLimits:r2>0.999ConductivitydetectorDRIFTANDNOISEPurewaterispumpedthroughthedetectorcellat1.0mL/min.Measurethebaselinenoiseoveranappropriateperiod.Limits:Noise:<2nSDrift:<20nS/hLINEARITYSolutions:5,10,25,50and100ppmsolutionsofNitrateinwaterforchromatographyR.Settings:Column:BackpressureloopEluent:DeionizedwaterFlowrate:1.0mL/minLimits:r2>0.999CoulometricdetectionFollowthemanufacturerinstructions.REFRACTIVEINDEXDETECTOR示差折光檢測器SignaltoNoiseratio,driftovertimeandlinearityaretheparametersproposedfortheperiodicandmotivatedcheckoftheRI(refractiveindex)detect0r.信噪比、隨時間的漂移和線性是提出的周期性和有動機(jī)的檢查的RI(折射率)探測器的參數(shù)。SIGNALTONOISERATIO信噪比Method1方法1p.18/25PA/PH/OMCL(11)04R6-Annex1toQualificationofEquipment:QualificationofLCEquipmentSolutions溶液?D-fructosesolutionat4.0mg/mL(e.g.dilute200.0mgfructosein20mLwithwaterforchromatographyR,add25mLacetonitrileRanddiluteto50.0mLwithwaterforchromatographyR)4.0mg/mLD-果糖溶液(例如，將200.0mg的果糖用20mL水稀釋，加入25mL乙腈，用水稀釋至50mL)Settings設(shè)置.Column:SpherisorbNH25^m,250x4.6mmorequivalent色譜柱：氨基柱，5^m,250x4.6mm或相當(dāng)?shù)腛ventemperature柱溫：38°CCelltemperature池溫：35°CFlowrate流速：1.0mL/minInjectionvolume進(jìn)樣體積:20pLMobilephase:0.253gsodiumdihydrogenphosphateRin220mLofwaterforchromatographyRand780mLofacetonitrileR流動相:0.253g磷酸二氫鈉在220mL水中+780mL乙腈Procedure程序：Afterequilibration,injectthreetimesablanksolutionofmobilephaseoveraruntimewherethesystemisstab