歐盟omcl儀器確認(rèn)1

QUALITYMANAGEMENTDOCUMENTPA/PH/OMCL(11)04

QUALIFICATIONOFEQUIPMENT

儀器確認(rèn)ANNEX1:QUALIFICATIONOFHPLCEQUIPMENT

附件1：HPLC儀器的確認(rèn)FulldocumenttitleandreferenceQualificationofEquipment文件全名和索引號Annex1:QualificationofHPLCequipmentPA/PH/OMCL(11)04Documenttype文件類型Guideline指南Legislativebasis立法基礎(chǔ)ThepresentdocumentwasalsoacceptedbyEAasrecommendationdocumenttobeusedinthecontextofQualityManagementSystemauditsofOMCLsDateoffirstadoptionMay2005首次發(fā)行日期Dateoforiginalentryintoforc(e June2005首次執(zhí)行日期Dateofentryintoforceofreviseddocument修訂后執(zhí)行日期1stJuly2011Previoustitles/otherreferencesThisdocumentreplacesdocumentPA/PH/OMCL(07)原文件名/其它索引號17DEFCustodianOrganisation保管機(jī)構(gòu)ThepresentdocumentwaselaboratedbytheOMCLNetwork/EDQMoftheCouncilofEuropeConcernedNetworkGEON相關(guān)網(wǎng)絡(luò)ANNEX1OFTHEOMCLNETWORKGUIDELINE

“QUALIFICATIONOFEQUIPMENT”O(jiān)MCL網(wǎng)絡(luò)指南“儀器的確認(rèn)”之附件1QUALIFICATIONOFHPLCEQUIPMENTHPLC儀器的確認(rèn)Introduction概述ThepresentdocumentisthefirstAnnexofthecoredocument"QualificationofEquipment”，anditshouldbeusedincombinationwithitwhenplanning,performinganddocumentingtheHPLCequipmentqualificationprocess.本文件是核心文件"儀器的確認(rèn)”第1個附件，在計劃、實施和記錄HPLC儀器的確認(rèn)過程時，應(yīng)將本文件與核心文件一起使用。ThecoredocumentcontainsthegeneralintroductionandtheLevelIandIIofqualification,commontoalltypeofinstruments,andthepresentannexcontainsHPLCinstrument-relatedrecommendationsonparameterstobecheckedandthecorrespondingtypicalacceptanceLimits,aswellaspracticalexamplesontheMethodologythatcanbeusedtocarryoutthesechecks.核心文件包括了第一級和第二級確認(rèn)的通用介紹，適用于所有類型的儀器，本附件包括了HPLC儀器相關(guān)的需要檢查的參數(shù)和相應(yīng)典型的可接受標(biāo)準(zhǔn)的推薦，以及可用于進(jìn)行這此檢查的實用方法學(xué)舉例。WhenqualifyingHPLCequipment,itshouldbenotedthatitisacceptabletocheckatLevelIIIandIVseveralofthementionedparametersatthesametimeinacombinedtestprocedure."overall”systemperformancetestgivinginformationonpeakareaprecision,retentiontimeprecision,gradientreproducibility,etc）.在進(jìn)行HPLC儀器確認(rèn)時，應(yīng)注意也可以對第三級和第四級幾個提到的參數(shù)采用聯(lián)合檢測程序同時進(jìn)行檢查（例如"全面”系統(tǒng)性能檢查，同時給出峰面積精密度、保留時間精密度、梯度重復(fù)性等的信息）TABLEIII表三LevelIII.Periodicandmotivatedinstrumentchecks第三級定期主動儀器檢查ExamplesofrequirementsforHPLCinstrumentsanddetectorsHPLC儀器和檢測器要求舉例Instrumentmodule儀器模塊Parametertobechecked需檢查的參數(shù)TypicaltoleranceLimits典型允差限度SolventdeliverysystemFlowrate流速±5%

溶劑傳送系統(tǒng) Proportioningaccuracyandprecision(gradienttest)酉己比準(zhǔn)確度和精密度(梯度測試)Proportioningripple酉己比波動±2W%Injector進(jìn)樣器Volumeprecision進(jìn)樣量精密度Carry-over殘留RSDW%seeAnnexI見附件1Autosampler自動進(jìn)樣器Thermostatting accuracy andprecision恒溫準(zhǔn)確度和精密度±3°COvenorcoolingdevice柱溫箱或冷卻裝置Thermostattingaccuracy恒溫準(zhǔn)確度土2°CUV/DADdetectorUV/DAD檢測器Linearity線性Wavelengthaccuracy波長準(zhǔn)確度r22±2nmFluorescencedetector熒光檢測器Wavelength accuracyexcitation波長激發(fā)準(zhǔn)確度Wavelengthaccuracyemission波長發(fā)射準(zhǔn)確度Sensitivity靈敏度±3nm±3nm±seeAnnexIElectrochemical Accuracyofthesignal信號準(zhǔn)detector電化學(xué)檢測器 確度Stabilityofthesignal信號穩(wěn)定性seeAnnexI見附件1seeAnnexI見附件1RIDdetector Signal/Noiseratio信噪比RID檢測器 Driftovertime飄移seeAnnexI見附件1mV/minCDdetector Signal/Noiseratio信噪比CD檢測器 Sensitivity靈敏度Driftovertime飄移Linearity線性seeAnnexI見附件1seeAnnexI見附件1seeAnnexI見附件1r>TABLEIV表四LevelIV.In-useinstrumentchecks第四級在用儀器檢查ExamplesofrequirementsforHPLCinstrumentswithUVorDADdetectors配備UV或DAD檢測器的HPLC儀器的要求舉例Parametertobechecked需檢查參數(shù)TypicaltoleranceLimits限度SystemsuitabilitycheckfortheMethodAccordingtoPh.Eur.orMAHdossieror方法系統(tǒng)適用性檢查validatedin-houseMethod根據(jù)歐洲藥典或MAH文件或公司內(nèi)部經(jīng)過驗證的方法Peakareaprecision(applicabletothemainpeaksinthetestsolution)峰面積準(zhǔn)確度(適用于供試液中主峰面積)RSDW%(unlessotherwiseprescribedinthesystemsuitabilityoftheMethod,.specificrequirementsfromPh.Eur.APImonographsorMAdossiers)除非在方法的系統(tǒng)適用性里有描述，例如特定要求，原料藥各論或MA文件Retentiontimeprecision保留時間精密度RSD土5%Carry-over(bycomparingconsecutivestandard(ofthesubstancebeingquantified)andblankinjections)殘留(通過比較連續(xù)(進(jìn)行定量檢測的物質(zhì))標(biāo)準(zhǔn)和空白進(jìn)樣)W%Signal/Noiseratio(tobeappliedforrelatedsubstancestest)信噪比(用于有關(guān)物質(zhì)檢測)AccordingtoPh.Eur.根據(jù)歐洲藥典ANNEXI附件1LevelIII.Periodicandmotivatedinstrumentchecks第三級定期性的主動儀器檢査ThisAnnexcontainspracticalexamplesoftestsandtheirassociatedtoleranceLimitsforseveralparametersrelatedtotheperformanceofthedifferentmodulesofaHPLC.本附件包括一些實用幾個與HPLC不同模塊相關(guān)的參數(shù)檢測舉例及其相應(yīng)的可接受限度。TheseexamplescanbeconsideredbytheOMCLsaspossibleapproachestoperformtheLevelIIIoftheequipmentqualificationprocess:"Periodicandmotivatedinstrumentchecks”.這些例子可以認(rèn)為是OMCL實驗室可能的進(jìn)行第三級儀器確認(rèn)的方法"周期主動儀器檢查”HPLCSOLVENTDELIVERYSYSTEM高壓液相溶劑傳輸系統(tǒng)ThefollowingtestsareproposedfortheperiodicandmotivatedcheckoftheHPLCsolventdeliverysystem:flowrateandgradienttest.以下測試作為HPLC溶劑傳輸系統(tǒng)周期性主動檢查：流速和梯度檢查FLOWRATE流速Materials:材料Volumetrieflaskof5or10ml5ml或10ml的容量瓶Calibratedchronometer校正過的計時計Settings:設(shè)置Mobilephase:degassedwater流動相：脫過氣的水Nocolumn(openend)*無柱(開放式)*Flowrate:adjustedbetweenandml/min流速：調(diào)節(jié)在到minIfhigh-pressuremixingsystemsareinstalled,thistesthastobedoneoneachsolventchannel.如果有安裝高壓混合系統(tǒng)，本測試應(yīng)在每一溶劑通道進(jìn)行。Forcertainequipment,.inthecaseoflowflowrates,thecheckwouldbeperformedbyusingacolumnorabackpressureregulator.對于一定的儀器，例如，流速較低時，采用一根柱子或背壓調(diào)整器進(jìn)行檢查Method方法：Settheflowrateatanappropriatelevelandmeasurethetimeneededtofillthevolumetrieflaskuptothemark.Recordthetimeneeded.將流速設(shè)定在一個合適的水平，測量將容量瓶裝滿至刻度的時間。記錄所用的時間。f=V/tf=(VX60)/1f measured flowrate[ml/min]測量的流速t elapsed timetofilluptomark[s]充滿至刻度所花的時間V volumeofthevolumetrieflask[ml]容量瓶的體積D=100X(f—F)/FD deviation[%]偏差F adjustedflowrate[ml/min]調(diào)節(jié)后的流速f measured flowrate[ml/min]測量的流速Limits限度：±5%GRADIENTCOMPOSITIONANDRIPPLE梯度組成和波動Settings:設(shè)置Stainlesssteelcapillary.2000xmminstalledinsteadofacolumn不銹鋼毛細(xì)管，例如，代替色譜柱安裝于儀器Detection:UV—Detectoradjustedto265nm檢測器：紫外檢測器調(diào)節(jié)至265nmMobilephaseA:degassedwater流動相A：脫氣水MobilephaseB:degassedwatercontaining%acetone流動相8：含%丙酮的脫氣水Flowrate:ml/min流速：分鐘Method方法：Thetestiscarriedoutinthefollowingwaybyusingagradientprogramdependingonthenumberofsolventchannelsandtheconfigurationofthesystem:按下列方法進(jìn)行測試，根據(jù)通道數(shù)量不同使用下列不同系統(tǒng)參數(shù)和梯度進(jìn)行洗脫A—BA—BandA—CA—C,A—BandB—Dtime[min]時間[分鐘]%mobilephaseA(water)%流動相A(水相)%mobilephaseB%流動相B(water-acetonemixture)(水-丙酮混合物)100090101090105050205050109030109001004001001000Startthetestbypumpingwaterforatleast10mintoequilibratethesystem.開始測試前，先用水平衡系統(tǒng)至少10分鐘。Thezero%valueatthestartofthetestisthebaseline.Allstepsaremeasuredatthebeginningofthehorizontalpartofthelineeitherbysoftwareormanuallyonthepaperprintusingaliner.Theheightofthe100%water/acetonemixtureisusedasthe100%valueinthefollowingcalculation.開始時的0%值是基線值。所有步驟均在開始時測量基線的水平部分，由軟件或手動在紙上用襯墊打印。100%水/丙酮混合物的高度在以下計算中作為100%值。%H=100*h/H%H calculatedcomposition計算組成h heightofthemeasuredline測量線的高度H heightofthe100%water/acetonemixtureline(mobilephaseB）100%水/丙酮混合物線的高度（流動相B）

d=%H—Gd deviation偏差G gradientcompositionadjusted[%acetone/watersolutionmixture=mobilephaseB]梯度組成調(diào)節(jié)【％丙酮/水溶液混合物=流動相B】Limits限度：Absolutedeviation:±2oftheadjustedvalue絕對偏差：調(diào)整值的±2Therippleofthegradientcompositionisthepercentageofnoiseofthe50%linefromthegradientprogram.梯度組分的波動為梯度程序噪聲基線的50%%R=100*N/h50%R ripple 波紋h height ofthe50%line50%的線高度50N heightofthenoiseline,measuredduring1minuteinthelinearregion噪聲線高度，在線性區(qū)域1分鐘測量的結(jié)果Limit限度s:W%HPLCINJECTOR進(jìn)樣器Volumeprecisionandcarry-overarethetestsproposedfortheperiodicandmotivatedcheckoftheHPLCinjector.建議在HPLC進(jìn)樣器周期性主動檢查中進(jìn)行體積精密度和殘留檢查VOLUMEPRECISIONANDCARRY-OVER體積精密度和殘留Solutions:溶液SolventA:methanol:waterR,60:40.溶液A：乙醇：水（試劑）=60：40Referencesolution（a）:dissolvemgmethyl-,ethyl-,andpropylparabeneinmlofsolventA.對照液（a）:取乙基、甲基和尼泊金丙酯溶于溶劑A中Referencesolution（b）:Dilutemlofreferencesolution（a）tomlofsolventA.對照液（b）：取對照液（a）用溶于溶劑A中Referencesolution（c）:Dilutemlofreferencesolution（b）tomlofsolventA.對照液（c）：取對照液（b）用溶于溶劑A中Settings:設(shè)置Column:Lichrospher100RP8,5um,125x4mm,withoutprecolumn色譜柱：Lichrospher100RP8,5um,125x4mm,無襯管Mobilephase:methanol:water=60:40流動相：乙醇：水=60：40Flowrate:ml/min流動速度：分鐘Detection:254nm檢測器：254nmInjectionvolume:20ul進(jìn)樣體積：20ulMethod方法：Injectionscheme:進(jìn)樣序列一 6xreferencesolution（b）對照液（b）6針一 lxreferencesolution（a）對照液（a）1針一 1xsolventA（blankinjection1）溶劑A（空白）1針lxreferencesolution(b)對照液(b)1針lxsolventA(blankinjection2)溶劑A(空白2)1針1xreferencesolution(c)對照液(c)1針Limits限度：Repeatabilityofpeakareas:Therelativestandarddeviationofthepeakareasofallpeaksinthechromatognmobtainedwiththereferencesolution(b)shouldbeW%.峰面積重復(fù)性：對照液(b)色譜圖中所有峰的峰面積相對標(biāo)準(zhǔn)偏差應(yīng)W%Carry-over:Thepercentageofthepeakareacorrespondingtopropylparabeneintheblankinjection1doesnotexceed%of10timesthepeakareaofthepropylparabenepeakinthechromatogramobtainedwiththereferencesolution(b)injectedaftertheblankinjection.殘留：尼泊金丙酯在空白圖譜中的峰面積不超過在空白后進(jìn)樣的對照液(b)中尼泊金丙酯的對應(yīng)峰面積的10倍的％Thepercentageofthepeakareacorrespondingtopropylparabeneinthereferencesolution(c)is-%ofthepeakareaofthepropylparabenepeakinthechromatogramobtainedwiththereferencesolution(b)injectedaftertheblankinjection.在對照液(c)中尼泊金丙酯對應(yīng)的峰面積為空白后進(jìn)樣的對照液(b)中尼泊金丙酯峰面積的。HPLCAUTOSAMPLER液相色譜儀自動進(jìn)樣器ThermostattingaccuracyandprecisioncanbetestedintheframeoftheperiodicandmotivatedcheckoftheHPLCAutosampler.HPLC自動進(jìn)樣器的周期主動檢查可以進(jìn)行溫度準(zhǔn)確度和精密度檢查THERMOSTATTINGACCURACY溫度準(zhǔn)確性Materials:材料Calibratedtemperatureprobe.校正過的溫度探頭Method方法：Selectatemperaturealongtheoperationalorrequiredtemperaturerangeoftheequipment.Waituntilthesystemisequilibrated.在可操作范圍或要求的溫度范圍內(nèi)選擇并設(shè)定一個溫度，等待系統(tǒng)平衡Bymeansofthecalibratedprobe,measuretheactualtemperatureintheautosamplerandcompareittotheselectedtemperature.采用校正過的溫度探頭，測量自動進(jìn)樣器的實際溫度，與設(shè)定的溫度進(jìn)行比較Repeatthesameprocedureatdifferentpre-selectedpointscoveringthetemperaturerange?在選定的溫度范圍內(nèi)不同溫度點重復(fù)相同的步驟Limits限度：Theactualtemperaturemaynotdiffermorethan±3°Cwithrespecttotheselectedtemperature.實際溫度與設(shè)定的溫度相比差值不超過±3°CTHERMOSTATTINGPRECISION溫度準(zhǔn)確性Materials:材料Calibratedtemperatureprobe.校正過的溫度探頭。Method方法：Selectatemperaturealongtheoperationalorrequiredtemperaturerangeoftheequipment.Waituntilthesystemisequilibrated.選擇一個操作溫度或要求的儀器溫度范圍，等待系統(tǒng)平衡。Bymeansofthecalibratedprobe,make“n”measurementsoverapresstablishedperiodoftime.Comparethemeanofthe“n”measurementstotheselectedtemperature.采用校正過的溫度探頭，在預(yù)設(shè)的時間內(nèi)進(jìn)行n次測量，比較n次測量值的平均值與設(shè)定的溫度。Limits限度：Theactualtemperaturemaynotdiffermorethan±3°Cwithrespecttotheselectedtemperature.實際溫度與設(shè)定的溫度之間差值不超過±3°CThermostattingaccuracyistheparametertestedinthisexampleofberiodicandmotivatedcheckoftheHPLCoven/coolingdevice.熱力學(xué)準(zhǔn)確性是在本例對HPLC柱溫箱/降溫設(shè)施周期主動檢查的參數(shù)THERMOSTATTINGACCURACY溫度準(zhǔn)確性Materials:材料Calibratedthermometer.校正過的溫度計Method方法：Setthecolumnoventemperatureto40°C,waitabout30minutestoequilibratethesystem,putacalibratedthermometerintotheovenandreadthetemperatureafter10minutes.設(shè)定柱溫箱溫度為40°C,等待約30分鐘平衡系統(tǒng)，將校正過的溫度計放入柱溫箱中，在10分鐘后讀取溫度。Limits限度：38-42°C.HPLCUV/DADDETECTOR紫外/DAD檢測器TheperiodicandmotivatedcheckoftheHPLCUV/DADdetectorcanbeperformedbytestingthelinearityandthewavelengthaccuracy.對于HPLC紫外/DAD檢測器的周期主動檢查，可以對線性和波長準(zhǔn)確性進(jìn)行檢查LINEARITY線性Solutions:溶劑配制Std.1:Ugcaffeine/1mlmethanolHPLCGrade標(biāo)準(zhǔn)2：Ug咖啡因/1ml色譜級甲醇Std.2:Ugcaffeine/1mlmethanolHPLCGrade標(biāo)準(zhǔn)2：Ug咖啡因/1ml色譜級甲醇Std.3:Ugcaffeine/1mlmethanolHPLCGrade標(biāo)準(zhǔn)3：Ug咖啡因/1ml色譜級甲醇Std.4:Ugcaffeine/1mlmethanolHPLCGrade標(biāo)準(zhǔn)4：Ug咖啡因/1ml色譜級甲醇Std.5:Ugcaffeine/1mlmethanolHPLCGrade標(biāo)準(zhǔn)5：Ug咖啡因/1ml色譜級甲醇Std.6:methanolHPLCGrade(blank)標(biāo)準(zhǔn)6：色譜級甲醇（空白）Std.5:weightomgcaffeineandfilluptomlwithmethanolHPLC標(biāo)準(zhǔn)5：稱取至咖啡因，用HPLC級別甲醇稀釋至Std.4:dilutemlofStd.5tomlwithmethanol標(biāo)準(zhǔn)4：取標(biāo)準(zhǔn)5溶液用甲醇稀釋至Std.3:dilutemlofStd.5tomlwithmethanol標(biāo)準(zhǔn)3：取標(biāo)準(zhǔn)溶液5用甲醇稀釋至Std.2:dilutemlofStd.3tomlwithmethanol標(biāo)準(zhǔn)2：取標(biāo)準(zhǔn)3溶液用甲醇稀釋至Std.1:dilutemlofStd.3tomlwithmethanol標(biāo)準(zhǔn)1：取標(biāo)準(zhǔn)3用甲醇稀釋至Settings:設(shè)置Column:RPT85um30-50x2,mmorcapillary2000mmxmmID色譜柱：RP-185um30-50x2,mmorcapillary2000mmxmmIDMobilephase:methanolHPLCGrade流動相：色譜級甲醇Oventemperature:40°C柱溫箱溫度：40°CFlowrate:ml/min(adjustedbyusing100%methanol)流速：min(用100%甲醇調(diào)整)Detection:273nm檢測波長：273nmInjectionvolume:20ul進(jìn)樣量：20ulMethod方法：Injectionscheme:進(jìn)樣序列2xblank1xStd.11xStd.21xStd.31xStd.41xStd.52針空白，標(biāo)準(zhǔn)1-5依序各一針Limits限度：r22Remark:Asthistestemploysdifferenttestsolutionstobeinjected,itcoversalsothecheckofcorrectpositioningvialsintheautosampler.注：由于此檢測中要注入不同溶液，因此也包括了自動進(jìn)樣器樣品瓶位置正確性的確認(rèn)WAVELENGTHACCURACY波長準(zhǔn)確度Iftherearebuilt—intestproceduresforthedeterminationandadjustmentofwavelengthaccuracy,followtheinstruetionsoftheinstrumentmanual.如果內(nèi)置檢測程序可以檢測和調(diào)節(jié)波長準(zhǔn)確度，則根據(jù)操作手冊進(jìn)行操作Inallothercasesusetheproceduredescribedbelow.在所有其它情況下采用以下程序Solutions:溶液DAD:caffeineStd.5fromthelinearitytesting線性測試中咖啡因標(biāo)準(zhǔn)溶液5號UV/VIS: ug/mlanthracenesolutioninwater蒽水溶液Settings:設(shè)置Mobilephase:15%acetonitrileinwater流動相：15%乙腈水溶液Column:RP18,5?m30—50x2,mmorcapillarymxmmID色譜柱：RP18,5um,30—50X或毛細(xì)管柱20mX內(nèi)徑Oventemperature:40°C柱箱溫度：40°CFlowrate:ml/min(adjustedbyusing15%acetonitrileinwater)流速：min(用15%的乙腈水溶液調(diào)節(jié))Detection:scanfrom230nmto290nm(DAD)檢測：自230至290nm掃描（DAD）Injectionvolume:20ul進(jìn)樣體積：20ulMethod方法：DAD:Inject20ulofthecaffeinesolutionandrecordthespectrum.Themaximumisat272nmandtheminimumat244nm.DAD：咖啡因溶液進(jìn)樣20ul,記錄色譜圖，最大吸收波長為272nm,最小為244nm.UV/VIS:fillthecellwiththeanthracenesolutionandchangethewavelengthfrom248to254nmin1nmsteps;recordthemaximumofabsorption.Thetheoreticalvalueis251nm.UV/可見光：向池中注入蒽溶液，將波長以lnm的步距從248變化至254nm,記錄最大吸收值。理論值為251nm。Limit限度s:W2nmHPLCFLUORESCENCEDETECTOR熒光檢測器ThefollowingthreeparametersareproposedfortheperformanceoftheperiodicandmotivatedcheckoftheHPLCfluorescentdetector:下面三個參數(shù)建議作為熒光檢測器性能周期自主檢查WAVELENGTHACCURACYEXCITATION波長激發(fā)準(zhǔn)確度Method:方法Rinseandfillthemeasuringcellwithde-ionizedwater用去離子水淋洗并充滿檢測池Adjusttheexcitationwavelengthto350nm.調(diào)節(jié)激發(fā)波長為350nmMeasuretheemissionandsubtract397nm(theoreticalvalue).測量發(fā)射波長減去397nm(理論值)Limits限度：±3nmWAVELENGTHACCURACYEMISSION波長發(fā)射準(zhǔn)確度Method：方法Rinseandfillthemeasuringcellwithde—ionizedwater用去離子水淋洗并充滿檢測池Adjusttheemissionwavelengthto397nm.調(diào)節(jié)發(fā)射波長為397nmMeasuretheexcitationandsubtract350nm(theoreticalvalue).測量激發(fā)波長減去350nm(理論值)Limits限度：±3nmSENSITIVITY靈敏度Solutions:溶液Quininesolutionconc.ug/ml(=15ppb)二水鹽酸喹啉溶液，濃度ug/ml(=15ppb)Thequininesolutionispreparedwiththefollowingmobilephase:dissolvegofpotassiumdihydrogenphosphateRandgofhexylamineRin700mlofwaterR,adjusttopHwithdilutephosphoricacidR,add90mlofacetonitrileRanddilutetomlwithwaterR.喹啉溶液用以下流動相制備：溶解磷酸二氫鉀和己胺于700ml試劑級水中，用稀磷酸調(diào)pH至，加90ml乙腈并用試劑級水稀釋至.Settings:設(shè)置ThechromatographicconditionsaresetaccordingtoPh.Eur."QuinineHCl”(01/2005:0018),test"Othercinchonaalkaloids”，withmodifiedflowrateandacetonitrileconcentration.色譜條件設(shè)定依據(jù)歐洲藥典"鹽酸喹啉”(01/2005:0018)，檢測"金雞納生物堿”，改變流速和乙腈濃度。Mobilephase:asabove流動相：同上Column:RP18,5um,250Xmm色譜柱：RP18,5um,250XmmFlowrate:ml/min流速：ml/minExcitationwavelength:350nm激發(fā)波長：350nmEmissionwavelength:397nm發(fā)射波長：397nmFlow-cellvolume:8ul(forthisexample,aWaters2475MultiFluorescentiedetectorwasused.Flow-cellvolumemayvarydependingontheinstrumentmanufacturer)流動池體積：8ul(此例中，使用的是Waters2475多熒光檢測器，流動池體積可能根據(jù)儀器廠商不同而不同)Method方法：Inject10ulofthequininesolutionandmeasurethepeakheight.進(jìn)樣喹啉溶液10ul,測量峰高度Inject10uloftheblankandmeasurethepeakheightofthenoise.進(jìn)樣空白溶液10ul,測量噪音峰高Dividethepeakheightofthequininesolutionby3timesthepeakheightofthenoise.將喹啉溶液峰高除以3倍噪音峰高Dividetheconcentrationofthequininesolutionbythepreviouslyobtainedfactor.將喹啉溶液濃度除以之前的系數(shù)Limits限度：WppbHPLCELECTROCHEMICALDETECTOR高效液相電化學(xué)檢測器AccuracyandstabilityofthesignalaretheproposedparameterstobetestedduringtheperiodicandmotivatedcheckoftheHPLCelectrochemicaldetector.電化學(xué)檢測器的信號準(zhǔn)確度和穩(wěn)定性是定期自主檢測中需要測定的參數(shù)ACCURACYANDSTABILITYOFTHESIGNAL信號準(zhǔn)確度和穩(wěn)定性Settings:設(shè)定Cellpotentialofadummycell:800mV空白池電壓：800mvRisetimefilter:s增加時間過濾:Range:nA范圍：Temperature:30°C溫度：30°CMethod方法：Accuracy:MeasuretheelectriecurrentandsubtractnA(theoreticalvalue)準(zhǔn)確度：檢測電流并減去(理論值)Stability:Measurethenoiseoveraperiodof5minutes穩(wěn)定性：連續(xù)5分鐘測量噪音Limits限度:Accuracy(cellcurrent):土nA準(zhǔn)確度(池電流)：土Stabilityofthesignal(Noise):max.2pAor20mV信號(噪音)穩(wěn)定性：最大2pA或20mVHPLCRIDDETECTOR高效液相RID檢測器SignaltoNoiseratioanddriftovertimearetheparametersproposedfortheperiodicandmotivatedcheckoftheHPLCRID(refractiveindex)detector.高效液相RID(折光率)檢測器的信噪比和對時間的漂移是定期自主檢測中需要測定的參數(shù)SIGNALTONOISERATIO信噪比Solutions溶液:Standardsolution:D-fructoseconcentratesolutionatmg/ml(dilutemgfructose+20mlwater+mlacetonitrileuptomlwithwaterforHPLC)標(biāo)準(zhǔn)溶液：D-左旋糖溶液ml(將左旋糖+20ml水+乙腈，用色譜純水稀釋至)Settings設(shè)置：Column:spherisorbNH2(orequivalent)250Xmmorother色譜柱：spherisorbNH2(或相當(dāng)?shù)?250Xmm或其他Oventemperature:38°C柱溫箱溫度：38°CFlowrate:ml/min流速：ml/minInjectionvolume:20ul進(jìn)樣量：20ulMobilephase:sodiumhydrogenphosphateRin220ml+780mlacetonitrile流動相：磷酸氫二鈉溶于220ml+780ml乙腈Method方法：Afterequilibration,injectthreetimesablanksolutionofmobilephaseoveraruntimewherethesystemisstable.Measurethebaselinenoiseoveranappropriateperiod.平衡后，以流動相為空白進(jìn)樣三針，系統(tǒng)穩(wěn)定運行適當(dāng)時間，測定適當(dāng)時長內(nèi)噪音基線Thebaselinenoiseisacceptedifthemeanheightofthethreereplicatesis<1000uV.可接受基線噪音：三次平行進(jìn)樣平均值〈1000UVTocalculatethesignaltonoiseratio,injectthreetimesasolutionoffructoseatmg/mlandcalculatethemeanofthethreereplicates.計算信噪比，左旋糖溶液ml進(jìn)樣三針，計算平均值Limits限度：S/N210限度：信噪比210DRIFTOVERADEFINEDPERIODOFTIME指定時長漂移Method：方法Calculatetheslopeoftheamplitudeofrandomvariationsinthedetector'ssignalover1minute.計算任意一分鐘信號隨機(jī)波動幅度的斜率Limits限度：土mV/min限度：土mV/minAlternatively,therequirementmaybeexpressedin△RI/minorin%offullscaleoftheselectedrange.也可以表達(dá)為△RI/min或者以選擇量程的全范圍％。HPLCCDDETECTOR高效液相色譜CD檢測器ThefollowingtestsareproposedtoperformtheperiodicandmotivatedcheckoftheHPLCCD(circulardichroism)detector.建議配備有CD(圓二圖譜)檢測器的HPLC進(jìn)行以下周期和主動檢查LINEARITYANDSIGNALTONOISERATIO線性和信噪比Solutions：溶液Referencesolution(a):dissolvemgD(-)pantolactoneinmlwater對照溶液(a):溶解D(-)泛酰內(nèi)酯于水中Referencesolution(b):dilutemlofreferencesolution(a)tomlwithwater對照溶液(b):取對照溶液(a)用水稀釋至Referencesolution(c):dilutemlofreferencesolution(a)tomlwithwater對照溶液(c)：取對照溶液(a)用水稀釋至.Referencesolution(d):dilutemlofreferencesolution(a)tomlwithwater對照溶液(d)：取對照溶液(a)用水稀釋至.Referencesolution(e):dilutemlo