欧盟OMCL仪器确认附件1HPLC确认之欧阳史创编.docx
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欧盟OMCL仪器确认附件1HPLC确认之欧阳史创编
OMCLNetworkoftheCouncilofEurope
QUALITYMANAGEMENTDOCUMENT
PA/PH/OMCL(11)04
QUALIFICATIONOFEQUIPMENT
仪器确认
ANNEX1:
QUALIFICATIONOFHPLCEQUIPMENT
附件1:
HPLC仪器的确认
Fulldocumenttitleandreference
文件全名和索引号
QualificationofEquipment
Annex1:
QualificationofHPLCequipment
PA/PH/OMCL(11)04
Documenttype文件类型
Guideline指南
Legislativebasis立法基础
ThepresentdocumentwasalsoacceptedbyEAasrecommendationdocumenttobeusedinthecontextofQualityManagementSystemauditsofOMCLs
Dateoffirstadoption
首次发行日期
May2005
Dateoforiginalentryintoforce
首次执行日期
June2005
Dateofentryintoforceofreviseddocument修订后执行日期
1stJuly2011
Previoustitles/otherreferences
原文件名/其它索引号
ThisdocumentreplacesdocumentPA/PH/OMCL(07)17DEF
CustodianOrganisation
保管机构
ThepresentdocumentwaselaboratedbytheOMCLNetwork/EDQMoftheCouncilofEurope
ConcernedNetwork
相关网络
GEON
ANNEX1OFTHEOMCLNETWORKGUIDELINE
“QUALIFICATIONOFEQUIPMENT”
OMCL网络指南“仪器的确认”之附件1
QUALIFICATIONOFHPLCEQUIPMENT
HPLC仪器的确认
Introduction概述
ThepresentdocumentisthefirstAnnexofthecoredocument“QualificationofEquipment”,anditshouldbeusedincombinationwithitwhenplanning,performinganddocumentingtheHPLCequipmentqualificationprocess.
本文件是核心文件“仪器的确认”第1个附件,在计划、实施和记录HPLC仪器的确认过程时,应将本文件与核心文件一起使用。
ThecoredocumentcontainsthegeneralintroductionandtheLevelIandIIofqualification,commontoalltypeofinstruments,andthepresentannexcontainsHPLCinstrument-relatedrecommendationsonparameterstobecheckedandthecorrespondingtypicalacceptanceLimits,aswellaspracticalexamplesontheMethodologythatcanbeusedtocarryoutthesechecks.
核心文件包括了第一级和第二级确认的通用介绍,适用于所有类型的仪器,本附件包括了HPLC仪器相关的需要检查的参数和相应典型的可接受标准的推荐,以及可用于进行这此检查的实用方法学举例。
WhenqualifyingHPLCequipment,itshouldbenotedthatitisacceptabletocheckatLevelIIIandIVseveralofthementionedparametersatthesametimeinacombinedtestprocedure(e.g.“overall”systemperformancetestgivinginformationonpeakareaprecision,retentiontimeprecision,gradientreproducibility,etc).
在进行HPLC仪器确认时,应注意也可以对第三级和第四级几个提到的参数采用联合检测程序同时进行检查(例如“全面”系统性能检查,同时给出峰面积精密度、保留时间精密度、梯度重复性等的信息)
TABLEIII表三
LevelIII.Periodicandmotivatedinstrumentchecks
第三级定期主动仪器检查
ExamplesofrequirementsforHPLCinstrumentsanddetectors
HPLC仪器和检测器要求举例
Instrumentmodule
仪器模块
Parametertobechecked
需检查的参数
TypicaltoleranceLimits
典型允差限度
Solventdeliverysystem
溶剂传送系统
●Flowrate流速
●Proportioningaccuracyandprecision(gradienttest)配比准确度和精密度(梯度测试)
●Proportioningripple配比波动
●±5%
●±2
●≤0.2%
Injector
进样器
●Volumeprecision进样量精密度
●Carry-over残留
●RSD≤1.0%
●seeAnnexI见附件1
Autosampler
自动进样器
Thermostattingaccuracyandprecision恒温准确度和精密度
●±3°C
Ovenorcoolingdevice
柱温箱或冷却装置
Thermostattingaccuracy
恒温准确度
●±2°C
UV/DADdetector
UV/DAD检测器
●Linearity线性
●Wavelengthaccuracy波长准确度
●r2≥0.999
●±2nm
Fluorescencedetector
荧光检测器
●Wavelengthaccuracyexcitation波长激发准确度
●Wavelengthaccuracyemission波长发射准确度
●Sensitivity灵敏度
●±3nm
●±3nm
●±seeAnnexI
Electrochemicaldetector电化学检测器
●Accuracyofthesignal信号准确度
●Stabilityofthesignal信号稳定性
●seeAnnexI见附件1
●seeAnnexI见附件1
RIDdetector
RID检测器
●Signal/Noiseratio信噪比
●Driftovertime飘移
●seeAnnexI见附件1
●0.1mV/min
CDdetector
CD检测器
●Signal/Noiseratio信噪比
●Sensitivity灵敏度
●Driftovertime飘移
●Linearity线性
●seeAnnexI见附件1
●seeAnnexI见附件1
●seeAnnexI见附件1
●r>0.999
TABLEIV表四
LevelIV.In-useinstrumentchecks
第四级在用仪器检查
ExamplesofrequirementsforHPLCinstrumentswithUVorDADdetectors
配备UV或DAD检测器的HPLC仪器的要求举例
Parametertobechecked
需检查参数
TypicaltoleranceLimits限度
●SystemsuitabilitycheckfortheMethod方法系统适用性检查
AccordingtoPh.Eur.orMAHdossierorvalidatedin-houseMethod
根据欧洲药典或MAH文件或公司内部经过验证的方法
●Peakareaprecision(applicabletothemainpeaksinthetestsolution)
峰面积准确度(适用于供试液中主峰面积)
RSD≤1.5%
(unlessotherwiseprescribedinthesystemsuitabilityoftheMethod,e.g.specificrequirementsfromPh.Eur.2.2.46,APImonographsorMAdossiers)除非在方法的系统适用性里有描述,例如EP2.2.46特定要求,原料药各论或MA文件
●Retentiontimeprecision保留时间精密度
RSD±5%
●Carry-over(bycomparingconsecutivestandard(ofthesubstancebeingquantified)andblankinjections)残留(通过比较连续(进行定量检测的物质)标准和空白进样)
≤0.2%
●Signal/Noiseratio(tobeappliedforrelatedsubstancestest)
信噪比(用于有关物质检测)
AccordingtoPh.Eur.
根据欧洲药典
ANNEXI附件1
LevelIII.Periodicandmotivatedinstrumentchecks
第三级定期性的主动仪器检查
ThisAnnexcontainspracticalexamplesoftestsandtheirassociatedtoleranceLimitsforseveralparametersrelatedtotheperformanceofthedifferentmodulesofaHPLC.
本附件包括一些实用几个与HPLC不同模块相关的参数检测举例及其相应的可接受限度。
TheseexamplescanbeconsideredbytheOMCLsaspossibleapproachestoperformtheLevelIIIoftheequipmentqualificationprocess:
“Periodicandmotivatedinstrumentchecks”.
这些例子可以认为是OMCL实验室可能的进行第三级仪器确认的方法“周期主动仪器检查”
HPLCSOLVENTDELIVERYSYSTEM高压液相溶剂传输系统
ThefollowingtestsareproposedfortheperiodicandmotivatedcheckoftheHPLCsolventdeliverysystem:
flowrateandgradienttest.
以下测试作为HPLC溶剂传输系统周期性主动检查:
流速和梯度检查
FLOWRATE流速
Materials:
材料
Volumetricflaskof5or10ml
5ml或10ml的容量瓶
Calibratedchronometer
校正过的计时计
Settings:
设置
Mobilephase:
degassedwater
流动相:
脱过气的水
Nocolumn(openend)*
无柱(开放式)*
Flowrate:
adjustedbetween0.5and3.0ml/min
流速:
调节在0.5到3.0ml/min
Ifhigh-pressuremixingsystemsareinstalled,thistesthastobedoneoneachsolventchannel.
如果有安装高压混合系统,本测试应在每一溶剂通道进行。
❆Forcertainequipment,e.g.inthecaseoflowflowrates,thecheckwouldbeperformedbyusingacolumnorabackpressureregulator.
对于一定的仪器,例如,流速较低时,采用一根柱子或背压调整器进行检查
Method方法:
Settheflowrateatanappropriatelevelandmeasurethetimeneededtofillthevolumetricflaskuptothemark.Recordthetimeneeded.
将流速设定在一个合适的水平,测量将容量瓶装满至刻度的时间。
记录所用的时间。
f=V/tf=(V×60)/t
f.................measuredflowrate[ml/min]测量的流速
t.................elapsedtimetofilluptomark[s]充满至刻度所花的时间
V...............volumeofthevolumetricflask[ml]容量瓶的体积
D=100×(f-F)/F
D...............deviation[%]偏差
F...............adjustedflowrate[ml/min]调节后的流速
f.................measuredflowrate[ml/min]测量的流速
Limits限度:
±5%
GRADIENTCOMPOSITIONANDRIPPLE梯度组成和波动
Settings:
设置
Stainlesssteelcapillarye.g.2000x0.12mminstalledinsteadofacolumn
不锈钢毛细管,例如2000X0.12mm,代替色谱柱安装于仪器
Detection:
UV-Detectoradjustedto265nm
检测器:
紫外检测器调节至265nm
MobilephaseA:
degassedwater
流动相A:
脱气水
MobilephaseB:
degassedwatercontaining0.5%acetone
流动相B:
含0.5%丙酮的脱气水
Flowrate:
1.0ml/min
流速:
1.0ml/分钟
Method方法:
Thetestiscarriedoutinthefollowingwaybyusingagradientprogramdependingonthenumberofsolventchannelsandtheconfigurationofthesystem:
按下列方法进行测试,根据通道数量不同使用下列不同系统参数和梯度进行洗脱
A-B
A-BandA-C
A-C,A-BandB-D
time[min]
时间[分钟]
%mobilephaseA(water)
%流动相A(水相)
%mobilephaseB%流动相B
(water-acetonemixture)(水-丙酮混合物)
0.0
100
0
0.1
90
10
10
90
10
10.1
50
50
20
50
50
20.1
10
90
30
10
90
30.1
0
100
40
0
100
40.1
100
0
Startthetestbypumpingwaterforatleast10mintoequilibratethesystem.
开始测试前,先用水平衡系统至少10分钟。
Thezero%valueatthestartofthetestisthebaseline.Allstepsaremeasuredatthebeginningofthehorizontalpartofthelineeitherbysoftwareormanuallyonthepaperprintusingaliner.Theheightofthe100%water/acetonemixtureisusedasthe100%valueinthefollowingcalculation.
开始时的0%值是基线值。
所有步骤均在开始时测量基线的水平部分,由软件或手动在纸上用衬垫打印。
100%水/丙酮混合物的高度在以下计算中作为100%值。
%H=100*h/H
%H............calculatedcomposition计算组成
h................heightofthemeasuredline测量线的高度
H...............heightofthe100%water/acetonemixtureline(mobilephaseB)
100%水/丙酮混合物线的高度(流动相B)
d=%H-G
d................deviation偏差
G...............gradientcompositionadjusted[%acetone/watersolutionmixture=mobilephaseB] 梯度组成调节【%丙酮/水溶液混合物=流动相B】
Limits限度:
Absolutedeviation:
±2oftheadjustedvalue绝对偏差:
调整值的±2
Therippleofthegradientcompositionisthepercentageofnoiseofthe50%linefromthegradientprogram.梯度组分的波动为梯度程序噪声基线的50%
%R=100*N/h50
%R............ripple波纹
h50.............heightofthe50%line50%的线高度
N...............heightofthenoiseline,measuredduring1minuteinthelinearregion噪声线高度,在线性区域1分钟测量的结果
Limit限度s:
≤0.2%
HPLCINJECTOR进样器
Volumeprecisionandcarry-overarethetestsproposedfortheperiodicandmotivatedcheckoftheHPLCinjector.
建议在HPLC进样器周期性主动检查中进行体积精密度和残留检查
VOLUMEPRECISIONANDCARRY-OVER
体积精密度和残留
Solutions:
溶液
SolventA:
methanol:
waterR,60:
40.
溶液A:
乙醇:
水(试剂)=60:
40
Referencesolution(a):
dissolve15.0mgmethyl-,ethyl-,andpropylparabenein100.0mlofsolventA.
对照液(a):
取15.0mg乙基、甲基和尼泊金丙酯溶于100.0ml溶剂A中
Referencesolution(b):
Dilute1.0mlofreferencesolution(a)to10.0mlofsolventA.
对照液(b):
取1.0ml对照液(a)用溶于10.0ml溶剂A中
Referencesolution(c):
Dilute1.0mlofreferencesolution(b)to100.0mlofsolventA.
对照液(c):
取1.0ml对照液(b)用溶于100.0ml溶剂A中
Settings:
设置
Column:
Lichrospher100RP8,5um,125x4mm,withoutprecolumn
色谱柱:
Lichrospher100RP8,5um,125x4mm,无衬管
Mobilephase:
methanol:
water=60:
40
流动相:
乙醇:
水=60:
40
Flowrate:
1.0ml/min
流动速度:
1.0ml/分钟
Detection:
254nm
检测器:
254nm
Injectionvolume:
20μl
进样体积:
20ul
Method方法:
Injectionscheme:
进样序列
— 6xreferencesolution(b)对照液(b)6针
— 1xreferencesolution(a)对照液(a)1针
— 1xsolventA(blankinjection1)溶剂A(空白)1针
— 1xreferencesolution(b)对照液(b)1针
— 1xsolventA(blankinjection2)溶剂A(空白2)1针
— 1xreferencesolution(c)对照液(c)1针
Limits限度:
Repeatabilityofpeakareas:
Therelativestandarddeviationofthepeakareasofallpeaksinthechromatogramobtainedwi