理想的实验室操作下.ppt

理想的實驗室操作(下),AnalyticalMethod分析方法Transfer,Verification,andValidation转移、确认和验证,MethodTransfer方法转移,Theprocesstoestablishdocumentedevidencethattheanalyticalmethodworksaswellinthereceivinglaboratoryasintheoriginatorslaboratory,orthetransferringlaboratory,iscalledAnalyticalmethodtransfer.用文件化的证明分析方法在接受实验室和原始实验室或者转移实验室一致的程序叫做方法转移,MethodTransfer方法转移,Regulatorytestingrequiresthatatransferofanalyticalmethodologyfromresearchthenthreeinjectionsforeachsolution?我可以配80%，100%，120%三个不同浓度的溶液，然后每个溶液各进三针吗？OrshouldIprepareninespikedsolutionswiththreein80%,threein100%,andthreein120%.Thenoneinjectionforeachsolution?或者是我配80%，100%，120%三个不同浓度的九个溶液，然后每个溶液各进一针？,Precision精密度,Precisionistheagreementbetweenasetofreplicatemeasurementswithoutassumptionofknowledgeofthetruevalue.精密度是一组测量值（不一定需要知道真实值）的重复性Precisionisusuallydefinedintermsofstandarddeviationorrelativestandarddeviation(%RSD).精密度通常用标准偏差或相对标准偏差来判定。,Precision精密度,Reproducibilitytheuseoftheanalyticalprocedureindifferentlabs.重现性分析程序在不同实验室中使用。Intermediateprecision(ruggedness)withinlabvariations,asondifferentdaysorwithdifferentanalystsorequipmentwithinthesamelab.中间精密度在同一实验室内变更条件，例如不同天数，或者是不同分析员，或者是同一实验室内的不同仪器。Repeatabilitytheuseoftheanalyticalprocedurewithinalaboverashortperiodoftimeusingthesameanalystandequipment重复性分析程序在实验室内同样的分析员和仪器在一小段时间内的使用。,ReproducibilityStudy重现性研究,Precision精密度,Repeatabilityshouldbeassessedusingaminimumofninedeterminationscoveringthespecifiedrangefortheprocedure.可重复性应该通过最少九个点涵盖整个指标范围来评估。Threeconc.andthreereplicatesofeachconc.三个浓度，每个浓度各重复三次。Aminimumofsixdeterminationsat100%ofthetestConcentration100%浓度的测试液最少六个点。,Specificity专属性,Theabilitytoassessunequivocallytheanalyteinthepresenceofcomponentswhichmaybeexpectedtobepresent,suchasimpurities,degradationproducts,andmatrixcomponents.评定被分析物与其他组分。例如杂质，降解产物，和空白基质的区分的能力,Specificity专属性,Forqualitativeanalysis(IDtest)theabilitytoselectbetweencompoundsofcloselyrelatedstructurethatarelikelytobepresentshouldbedemonstrated对于定性分析区分可能出现的相近结构的的组分的能力应该被证明。Forquantitativeanalysis(Assay)Demonstratethattheprocedureisunaffectedbythepresenceofimpuritiesorexcipients.Thismaybedonebyspikingwithimpurities.对于定量分析（含量）证明分析程序不受可能出现的杂质和赋形剂的影响，这个可以通过加入杂质来证明。,Specificity专属性,Positiveresultstheretentiontimeoftheanalytematcheswiththatofreferencestandard.阳性结果被分析物的保留时间和对照标准品一致。NegativeresultsReagentblankand/ormethodblankhasnopeakattheretentiontimeofanalyte.阴性结果试剂空白或者方法空白在被分析物保留时间未出峰,Specificity(PeakPurity)专属性（峰纯度）,Peakpuritytestmaybeusefultoshowthattheanalytechromatographicpeakisnotattributabletomorethanonecomponent.峰纯度测试表明被分析物色谱峰不是多个组分形成的非常有用。Peakpuritytestscanbeevaluatedwith峰纯度测试可以用下面来评价：Thespectraofphotodiodearraydetector二极管阵列MassSpectrometry质谱,LimitofDetection检测限,Thedetectionlimitisthelowestamountofanalyteinasamplewhichcanbedetectedbutnotnecessarilyquantitated,underthestatedexperimentalconditions检测限是在指定的实验条件下，被分析物能够检测出来的，不需要定量的最小量。Typicallyacceptablesignaltonoiseratiosis3:1.可接受标准一般是信噪比为3：1.,LimitofQuantitation定量限,Thequantitationlimitofanindividualanalyticalprocedureisthelowestamountofanalyteinasamplewhichcanbequantitativelydeterminedwithsuitableprecisionandaccuracy.定量限是指样品中被分析物满足精密度和准确度要求的最低量，,LimitofQuantitation定量限,Thequantitationlimitisaparameterofquantitativeassaysforlowlevelsofcompoundsinsamplematrices,andisusedparticularlyforthedeterminationofimpuritiesand/orDegradationproducts.定量限是样品中低含量组分定量计算时的一个参数，主要用来检测样品中的杂质和/或者降解产物。,LimitofQuantitation定量限,Thequantitationlimitshouldbeshowntobesufficientlylowbytheanalysisofsampleswithknownconcentrationsofanalyteaboveandbelowthequantitationlevel.通过分析已知的在定量限上下的样品浓度以确定定量限，定量限应该尽量低，Typicallyacceptablesignaltonoiseratiois10:1atwhichtheanalytecanreliablybequantitated.通常被分析物可以定量，可接受信噪比为10：1,LinearityandRange线性和范围,Determinelinearityoveraminimumoffiveconcentrations通过至少五个浓度来确定线性Assayofdrugsubstance:80%120%原料药含量：80%-120%Determinationofanimpurity:50%120%ofthespecification杂质：指标的50%120%Contentuniformity:Aminimumof70%130%ofthetestconcentration含量均匀度：测试溶液的70%130%Dissolutiontesting:20%throughoutthespecifiedrangeofthedissolutiontest.溶出测试：溶出测试指标范围的20%,Robustness耐用性,Robustnessisthecapacityofamethodtoremainunaffectedbysmall,deliberatevariationsinmethodparameters;itisameasurementofthereliabilityofamethod.耐用性是分析方法参数发生微小的，有意的改变，分析方法仍然有效的能力，反应了分析方法的可靠性。,SystemSuitability系统适用性,Systemsuitabilityisthecheckingofasystemtoensuresystemperformancebeforeorduringtheanalysisofunknowns.系统适用性是在分析未知样品前或者中间确保系统性能的系统检查。Itsparametersareestablishedasadirectresultofruggednessandrobustnessstudies系统适用性参数通过重现性和耐用性研究结果确定。,SystemSuitability系统适用性,Theoreticalplatecount(N),peaktailingfactor(T),resolution(R),andreproducibility(%RSD)aredeterminedfromreplicateinjectionsofastandardcomparedwithmethodspecifications.通过测定理论塔板数，峰拖尾因子，分离度，标准品重复进样重现性和方法标准相比较，IfRSDisstatedlessthan2.0%,fivereplicatesshouldbeused.如果RSD规定了小于2.0%，重复进样五次。IfRSDisstatedgreaterthan2.0%,sixreplicatesshouldbeused.如果RSD规定了大于2.0%，重复进样六次。,QCSamplesQC配样,TwoseparateweightofReferenceStandards(RS)andusedoneofthemasaninitialconcentrationverification,e.g.:称取两个不同重量的工作标准品，用其中一个作为初始浓度确认，例如：20mg100.0mL10mg50.0mLTwoseparateweightofSamples,e.g.:两个不同重量的样品20.2345mg100.0mL21.8765mg100.0mLSpikesolution(equalvol.ofRSandSample)配样：工作标准品和样品应该等体积,液相序列,DataRecording数据记录,Howisrawdatacontrolled原始数据怎么控制Howisdatamaintained/stored原始数据怎么保持和存储Boundbooks/Notebooks装订成册Sequentiallynumberedforms(Worksheets)数据表格Logbooks使用记录LIMS实验室信息管理系统Traceability可追踪性,MethodValidationReport方法验证报告,Asummaryreportshouldinclude:一个完整的报告应该包括：Instrument/Equipmentused使用的仪器/设备Reagents,Chemicals,andReferenceStandard(s)used试剂，化学品，工作标准品Numberofsamplestested测试的样品数Summaryofeachvalidationtest每个验证测试的概要AcceptanceCriteria可接受标准ReviewandApproval数据审核和批准,ExamplesofMethodsthatRequireValidationDocumentation需要验证文件的方法举例,ChromatographicMethodsHPLC,GC,TLC,LC/MS,GC/MS,etc.色谱方法液相，气相，点板，液质连用，气质连用，等等。PharmaceuticalAnalysisinsupportofCMCBioanalyticalAnalysisinsupportofPK/PD/ClinicalStudiesSpectrophotometricMethodsUV/Vis,IR,NIR,AA,NMR,XRD,MSetc.光谱学方法紫外可见，红外，近红外，原子吸收，核磁，多晶衍射，质谱,ExamplesofMethodsthatRequireValidationDocumentation需要验证文件的方法举例,CapillaryElectrophoresisMethodsZone,IsoelectricFocusing,Isotachophoresis,etc.毛细管电化学ParticleSizeAnalysisMethodsLaser,Microscopic,Sieving,etc.粒径分析方法激光粒度，显微镜，筛分，等等。DissolutionMethods溶出方法TitrationMethods滴定方法AutomatedAnalyticalMethods自动化的分析方法,ISO17025ValidationRequirementISO17025验证要求,Thevalidationshallbeasextensiveasisnecessarytomeettheneedsofthegivenapplicationorfieldofapplication.验证应尽可能的广泛，要满足特定应用或应用领域的需要。Thelaboratoryshallrecordtheresultsobtained,theprocedureusedforthevalidationandastatementastowhetherthemethodisfitfortheintendeduse.实验室应该记录获得的结果、验证的程序和一个关于方法是否适用的综述。,MethodValidation方法验证,Mostofthemethods(ifnotall)shownintheUSPmonographsareNOTastabilityindicatingmethod!USP专论中的大部分方法（如果不是所有的）并不是一个稳定性研究的方法Whendevelopandvalidateananalyticalmethod,oneshouldinclude“ForcedDegradationStudies”toshowthemethodisastabilityindicatingmethod!当开发和验证一个分析方法时，应该包括“强制降解研究”以证明方法是一个稳定性研究的方法。,ForcedDegradationStudies强制降解研究,HighTemperature(50to60)高温(50to60)Humidity(70to80%RH)高湿(70to80%RH)AcidInducedDegradation(HCl0.1N)酸降解(HCl0.1N)BaseInducedDegradation(NaOH0.1N)碱降解(NaOH0.1N)HydrogenPeroxideInducedDegradation(3to30%)过氧化氢氧化降解(3to30%)IntenseUV/VisibleLightandSunLight强烈紫外/可见光和日光降解Withintentofcreating10to30%degradation产生10%到30%降解,