分析方法验证程序

1、SOP标题SOP Title分析方法验证/确认Analytical Method Validation / Verification标准操作规程Standard Operating ProcedureSOP标题SOP Title分析方法验证/确认Analytical Method Validation / VerificationSOP编号SOP No.SOP-ARD-002版本Rev No.00执行日期Effective Date复审日期Review Date DATE d MMMM yyyy 31 August 2015Page SOP审核和批准 SOP Review and Approv

2、al职责Responsibility签名Signature日期Date打印名Printed Name职位Title起草人Created by 分析副主任Analytical Associate Director 复核Reviewed by分析主任Analytical Director批准人Approved by 副总经理Vice President批准人Approved byQA经理QA Manager目的Purpose本规程的目的是为在XXX有限公司进行的验证研究制定一个程序，包括分析方法验证过程，分析方法的确认，文件记录，审计，原始数据归档以及验证/确认文件放行。范围Scope本标准操作程

3、序适用于XXX有限公司产品研发部门GMP分析实验室，采用色谱技术对最终成品和API（如适用）进行的所有分析方法的验证/确认。本标准操作程序也可作为XXX有限公司产品研发部门分析研发实验室，采用其他分析技术进行其他任何方法验证/确认的参考。c. 职责Responsibility分析员Analyst负责准备验证方案Responsible for preparation of validation protocol负责进行方法验证/确认Responsible for execution of method validation / method verification负责准备验证报告/确认报告Re

4、sponsible for preparation of method validation report / method verification report主管小组/指定人员Group In-charge / Designee负责复核和审批验证方案Responsible for review & approval of validation protocol负责方法验证/确认活动的分配、监督以及验证/确认数据的复核Responsible for allotment & monitoring of method validation / verification activity and

5、 review of validation / verification data负责验证报告/确认报告的复核和审批Responsible for review & approval of validation report / verification report质量保证Quality Assurance 负责验证报告/确认方案的复核和审批Responsible for review & approval of validation report / verification protocol负责验证报告/确认报告的复核和审批Responsible for review & approva

6、l of validation report / verification report注册Regulatory 负责验证/确认方案的复核Responsible for review of validation/verification protocol.简写和定义Abbreviations & Definitions序号S. No.简写Abbreviations定义Definitions4.1验证（全验证）Validation(Full Validation)分析规程的验证是，通过实验室研究，确定该规程的工作特性达到了预定分析用途要求的过程。Validation of an analytic

7、al procedure is the process by which it is established, by laboratory studies, that the performance characteristics of the procedure meets the requirement for the intended analytical application. 4.2准确度Accuracy分析规程的准确度是由该规程得到的测试结果与真实值的接近程度。The accuracy of an analytical procedure is the closeness of

8、the test result obtained by that procedure to the true value.4.3精密度Precision分析规程的精密度是当该分析规程单独分析均质样品的多个样本时，若干检验结果的一致程度。The precision of an analytical procedure is the degree of agreement among individual test results when the procedure is applied repeatedly to multiple samplings of a homogenous sampl

9、e.4.4专属性Specificity专属性是指是当待分析物含有预期会有的其他组分（杂质，降解产物，基质成分等），准确可靠地评估待分析物的能力。The specificity is the ability to access unequivocally the analyte in the presence of components that may be expected to be present (impurities, degradation product, matrix component etc).4.5检测限Detection Limit指在规定的试验条件下，样品中可被检测到

10、的待分析物的最小数量，但是无需定量。It is the lowest amount of analyte in a sample that can be detected, but not necessarily quantitated, under stated experimental condition.4.6定量限Quantification Limit规定试验条件下，能够以可接受的精密度和精确度进行测定的样品中待分析物的最低量。It is the lowest amount of analyte in a sample that can be determined with acce

11、ptable precision and accuracy under stated experimental condition.4.7线性Linearity分析方法的线性度是通过直接或者明确给出的数学转换而间接地，得出与特定范围内的样品中待分析物浓度呈比例关系的测试结果的能力。The linearity of an analytical procedure is its ability to elicit test results that are directly or by well-defined mathematical transformation, proportional t

12、o the concentration of the analyte in samples with in a given range i.e. linearity of the relationship of concentration and assay measurement.4.8范围Range分析规程的范围是分析物的较高浓度和较低浓度（含）之间的区间，已经证实在此区间内，使用该规程进行测定具有适当水平的精密度、准确度、和线性。The range of an analytical procedure is the interval between upper and lower lev

13、el of analyte (including these levels) that have been demonstrated to be determined with a suitable level of precision, accuracy and linearity.4.9耐用性Robustness分析规程的耐用性是规程文件中列出的操作参数在微小、故意的变更中不受影响的能力的衡量单位，并在日常使用中提供了其适用性的指标。The robustness of an analytical procedure is a measure of its capacity to remai

14、n unaffected by small but deliberate variation in procedural parameter and provide an indication of its suitability during normal usage.4.10确认Verification对于药典测试方法确认过程是在实际使用条件下，评估对于特定的药物和/或药品，药典方法是否能够满足预期使用标1。The verification process for compendial test procedures is the assessment of whether the com

15、pendial procedure can be used for its intended purpose, under the actual conditions of use for a specified drug substance and/or drug product matrix. 4.11再验证（部分验证）Re-Validation(Partial Validation)一个分析程序重新验证的过程是重新建立分析方法，确保该分析方法在工艺、处方和方法改变的情况下，依然可以满足预期的分析应用的要求。 Re-validation of an analytical procedure

16、 is the process to re-establish the performance characteristics of the analytical procedure that can meet the requirement for the intended analytical application, considering any change in process / formulation / method etc.4.12再确认Re-Verification对于药典测试方法重新确认的过程，是评估药典方法在工艺、处方和方法改变的情况下，药典测试方法依然能满足预期使用

17、目标。Re-verification process for compendial test procedures is the re- assessment of whether the compendial procedure can be used for its intended purpose, considering any change in process / formulation / method etc.程序Procedure为了启动方法验证，相关人员或者指定人员应该起草验证方案。验证方案应该包含在验证过程中所有需要研究的参数和接受标准。相关主管或者指定人员应该对验证方案

18、进行复核。To initiate method validation, a validation protocol should be filled by the concerned person / designee. The validation protocol should contain all the parameters required to be studied during the validation along with acceptance criteria. Validation protocol should be checked by the concerned

19、 group head / designee. 备注：对于方法确认，不要求准备方案。Note: There is no requirement to prepare any protocol for method verification activity.验证方案应该有QA给定的唯一编号。The validation protocol should have a unique number given to it by the quality assurance. 验证方案应该由宣泰主管或者指定人员批准后，接着由QA和注册（如适用）批准。The validation protocol sho

20、uld be approved at Sinotherapeutics by group-in-charge / designee followed by approval from a representative of QA & Regulatory (if applicable).验证数据应该按照批准的验证方案在实验记录本（专门用于验证实验/观察）中记录。但是确认数据可以记录在方法研发记录本中，对于通用的药典测试方法不需要进行确认（如干燥失重、炽灼残渣、各种化学测试流程如酸度、简单的仪器测定如pH测定）。Validation data should be recorded in labo

21、ratory note book (dedicated for validation experiments/observations only) against each approved validation protocol. However, Verification data can be recorded in method development notebook. Verification is not required for basic compendial test procedures that are routinely performed (e.g. loss on

22、 drying, residue on ignition, various wet chemical procedures such as acid value, and simple instrumental determinations such as pH measurements). 在实验进行过程中或者实验完成后，来自方案的任何偏差应该被评估。任何来自方案的增补或变更应该在验证方案的合适章节中提到。During the execution of experimental work or after its completion, any deviation from the prot

23、ocol should be justified. Any additions or changes from the protocol should be mentioned under appropriate section in validation protocol.如果在之后的时间里，发现批准的方案中有一些小的差异或错误，如印刷或输入错误，主管或指定人员应该用“勘误表”说明并纠正这些错误。If at a later date minor discrepancies or errors are noticed in the approved validation protocol, s

24、uch as typographical and transcriptional errors, an ERRATUM stating these errors, along with the correction, should be prepared by the group in-charge/designee.在验证研究过程中，如果需要做额外的一些实验/参数，但它们不是主验证方案的一部分，就需要再产生一个补充方案（以及相应的编号），补充的实验将成为验证报告的一部分。During the course of validation studies, if any additional ex

25、periment / parameter needs to be done which is not part of the main validation protocol, an addendum protocol (and corresponding number) need to be taken, if that additional experiment will be part of validation report. 备注：补充验证方案可在最终的验证报告批准前的任何时间产生。最终的验证报告应包括来自于主验证方案和补充验证方案(如果有)的数据。Note: Addendum va

26、lidation protocol can be taken any time before the approval of final validation report. The final validation report should include the data generated against the main validation protocol and addendum protocol(s), if any.以下参数在验证活动中应该被考虑（但不限于这些）（方法验证/确认参数的选择，是基于个别技术/测试的特殊要求。但是，基于一些发现，可追加研究一些验证参数，这在任何情

27、况下都适用）。Following parameters should be considered (but not limited to) during validation activity / verification (Method validation parameters can be selected based on the specific requirements for particular technique / test. However, based on the observation, additional validation parameters can be

28、 studied, wherever applicable). 编号S. No.参数Parameters验证Validation确认Verification再验证Re-validation再确认Re-verification早期阶段验证Validation inEarly phase1准确度AccuracyYesYesYesYesYes2方法精密度Method PrecisionYesYes / NoYesYesYes3中间精密度Intermediate PrecisionYesNoNoNoNo4专属性/强制降解Specificity / Forced degradation*YesYesYe

29、s / NoYes / NoYes / No5线性/范围Linearity / RangeYesYes / NoNoNoYes6检测限Detection Limit (DL)Yes / NoYes / NoNoNoYes / No7定量限Quantitation Limit (QL)Yes / NoYes / NoNoNoYes / No8耐用性Robustness*YesNoNoNoNo9分析溶液稳定性Stability in analytical solutionYesYesYesYesYes* 强制降解和耐用性数据可以借用任何适用的、已有的方法开发研究的数据。Forced degrada

30、tion & Robustness data can be borrowed from method development studies, wherever applicable / if available.单个验证方案中的接受标准可参考附件-A中规定的接受标准，最终在批准的验证方案中的规定接受标准将会取代SOP中规定的接受标准。The acceptance criteria specified in appendix-A can be used as reference to define the acceptance criteria in the individual valida

31、tion protocol. Finally, the acceptance criteria specified in approved validation protocol will supersedes the acceptance criteria specified in this standard operating procedure.在实验工作和计算完成后，分析员应该准备验证/确认报告草案。On completion of the experimental work and calculations, a draft validation report / verificat

32、ion report should be prepared by analyst.主管/指定人员以及之后的QA，应该从准确度和完整性角度审核验证/确认报告草案和方法验证研究的原始记录。The draft validation report / verification report and the raw data of the method validation studies should be reviewed for accuracy and adequacy by group-in-charge / designee, followed by quality assurance.如果

33、有基于质量调查结果的纠正措施，应该在给予关注，并准备验证报告。Corrective actions, if any, based on the quality assurance findings should be taken care of and a final validation report should be prepared. 分析部门主管/指定人员应该首先审批报告，然后由QA/指定人员审批。报告应该有QA分发的独立编号。The report should be approved by Head Analytical Research (or R&D) / Designee,

34、followed by Quality Assurance / Designee. The validation report should have a unique number, as given by Quality Assurance.归档最终报告的原件以及原始记录，销毁验证报告草案。The final original copy of the report along with the raw data should be archived. The draft report should not be retained.如果在最终报告签署后，要求产生额外的数据而不是更新原来已经存

35、在的数据，应进行额外的实验工作，并准备一份原报告附录，放行额外产生的数据。If at a later date additional data other than the already existing data is required to be generated, additional experimental work should be carried out and an addendum to the original validation report should be prepared and issued.如果报告批准后，发现一些小的差异和错误，如印刷或输入错误（不超

36、过5个），应准备“勘误表”说明和纠正这些错误。如果印刷或输入错误超过5个或者需要对一些现存数据的报告进行修正，那就需要准备和放行修订版本的报告或者补充报告。If at a later date minor discrepancies or errors are noticed in the approved report report, such as typographical and transcriptional errors (not more than five in number), an ERRATA stating these errors, along with the co

37、rrection, should be prepared. If the typographical and transcriptional errors are more than five or there is any modification in the reporting of an existing data, a revised v report or addendum report should be prepared and issued. 原报告修订的原因应该清晰地在修订的报告中阐述。修订报告和原始记录应该交给QA审计。The reason for revision of

38、 the original report should be clearly stated in the introduction of the revised report. The revised validation report and the raw data should be submitted to quality assurance for audit. 批准的报告，以及原始记录、补充文件、勘误表和升级版本，都应该归档。All the approved reports along with raw data, addendum, erratum and version, wh

39、erever applicable, should be archived.如有需要，每份报告的复印件，应该同有关的生产厂家和注册部门分享。勘误表、补充文件和修订的报告以应该同原始验证报告的接受人员分享。A copy of each of the report should be shared with the concerned manufacturing location(s) and Regulatory, if required. The errata, addendum and revised v reports should be shared with the recipient

40、s of the original report, wherever applicable.分析方法的方案和报告应该保存至产品生命周期结束再加一年。Analytical method protocol and report(s) should be preserved up to a minimum period of Product life cycle + one year.附件Appendices附件编号Appendix No.附件名称Appendix NameF-ARD-002-A推荐的验证实验和相应的接受标准指导原则Recommended guideline for validati

41、on experiments and corresponding acceptance criteria参考资料（如有）References 联邦食品,药品和化妆品法案章节501 Federal Food, Drug & Cosmetic act Section 501药品生产质量管理规范21CFR 211.194 (a), 21CFR 211.194 (a) (2)cGMP Practice Regulations 21CFR 211.194 (a), 21CFR 211.194 (a) (2) 国际药品注册协调会议: Q2 (R1)International Conference on H

42、armonization: Q2 (R1)美国药典普通章节 法规方法的验证USP General Chapter Validation of Compendial Procedures美国药典通则药典方法的确认USP General Chapter Verification of Compendial Procedures变更历史History of Changes版本编号 Version No.替换版本编号Superseded Version No.变更描述Change(s) Made00N/A不适用/新文件Not Applicable / New Document Page of 10推荐

43、的验证实验指导原则和相应的接受标准Recommended guidelines for validation experiment and corresponding acceptance criteria含量/含量均匀度/混合均匀度方法验证指导原则Validation Guidelines for Assay / Content Uniformity / Blend Uniformity验证性能特征Validation performance Characteristics实验设计Experiment Design推荐的接受标准RecommendedAcceptance Criteria线性

44、Linearity至少配制5份样品目标浓度附近（至少包括70-130%）的不同浓度水平的药物溶液（无基质成分）。Solution of drug substances (without matrix component) to be prepared at a minimum of 5 different concentration level of the target test concentration (covering the minimum range of 70% - 130%) of sample. 用线性溶液的响应（面积，峰高，吸收值等）对其含量（浓度），画出线性曲线。报告相

45、关系数，回归线%的截距和斜率。Plot the linearity curve from the response (area, height, absorbance etc) versus amount (concentration) of the linearity solutions. Report the correlation coefficient, % y-intercept & slope of regression line.备注Note:混合均匀度/含量均匀度可能会要求验证一个更宽的范围。Blend uniformity / content uniformity may r

46、equire a wider range to validate.相关系数 0.99Correlation coefficient 0.99准确度Accuracy 配制含辅料和药物（纯度已知）的溶液，至少3个浓度水平（每个水平3份），覆盖的最小范围为样品目标测试浓度的70%-130%。计算回收率。Prepare a solution of placebo & drug substance (of known purity) at a minimum of three concentration level (each in triplicate), covering the minimum r

47、ange of 70%-130% of target test concentration and calculate the % recovery.备注Note: 混合均匀度/含量均匀度可能会要求验证一个更宽的范围。Blend uniformity / content uniformity may require a wider range to validate.每个浓度水平的每份样品的%回收率应在95 %- 105 %范围内% Recovery of individual sample at each level should be within 95 % to 105 %.精密度Pre

48、cision方法精密度（重复性）Method Precision (Repeatability)根据规定的流程，分别重复配制样品溶液6次（N=6），计算结果（%标示量）的%RSD 。Prepare sample solution six times (N=6) individually as per the defined procedure and calculate the % RSD from results (% against label claim)% RSD (N=6) 2 %中间精密度（耐用性）Intermediate Precision (Ruggedness)根据规定的流程

49、，在不同天，用不同的仪器，由不同的分析员，分别重复配制样品溶液6次（N=6），计算结果（%标示量）的%RSD 。Prepare sample solution six times (N=6) individually as per the defined procedure on another day using another instrument and performed by another analyst. Calculate the % RSD from results (% against label claim).% RSD (N=6) 2 % 总%RSD（方法精密度和中间精

50、密度）（N=12） 2 %The overall (method precision & intermediate precision) % RSD (N=12) 0.98Correlation coefficient 0.98准确度Accuracy 配制含辅料和药物（纯度已知）的溶液，至少3个浓度水平（每个水平3份），覆盖的最小范围为最低溶出质量标准（20%）至目标测试浓度的120%。计算回收率。至少配制3个浓度水平（每个水平3份）的药物（纯度已知）和辅料的混合溶液，范围至少覆盖样品目标测试浓度的70%-130%。Prepare a solution of placebo & drug su

51、bstance (of known purity) at a minimum of three concentration level (each in triplicate), covering the minimum range from lowest dissolution specification (20%) to 120% of target test concentration and calculate the % recovery.备注Note: 所有回收率实验的操作和处理，使用到的所有滤膜、注射器、样品处理步骤和稀释，都需要模仿实际的溶出样品。Handle & proces

52、s all recovery experiments using all the filters, syringes, sample handling procedure & dilutions that are used to mimic the actual dissolution sample.每个水平，每个样品的%回收率应该在90%-110%范围内。% Recovery of individual sample at each level should be within 90 % to 110 %.精密度Precision方法精密度（重复性）Method Precision (Rep

53、eatability)根据规定的步骤，使用6份 （N=6）单独的样品/辅料中加入药物，进行溶出实验。Perform the dissolution using six (N=6) individually units / placebo spiked with drug substances as per the defined procedure.在规定的时间点，对%标示量的计算%RSD.Calculate the % RSD from results of % against label claim at specification time point.% RSD 0.95Correla

54、tion coefficient 0.95定量限Quantification limit将要求定量限水平的线性溶液，进样6次。Inject the linearity solution prepared at required quantification level and inject 6 times.确认在要求的定量限水平，满足接受标准。用信号和噪音的比值，计算和报告实际的定量限。Verify the acceptance criteria are met at the required quantification level and calculate & report the ac

55、tual quantification level using signal to noise ratio.RSD 10:1Individual signal to noise ratio 10:1检测限Detection limit将要求检测限水平的线性溶液，进样6次。Inject the linearity solution prepared at required detection level and inject 6 times.确认在要求的定量限水平，满足接受标准。用信号和噪音的比值，计算和报告实际的定量限。Verify the acceptance criteria are me

56、t at the required quantification level and calculate & report the actual detection level using signal to noise ratio.RSD 3.3:1Individual signal to noise ratio 3.3:1准确度Accuracy 最好用药物产品（或辅料）和已知杂质，配制溶液，至少3个浓度水平（每个水平3份），覆盖的最小范围，要求从定量限（杂质报告限），质量标求准限/水平至质量标准限以上（约150%）。计算各个溶液的%回收率。Prepare a solution of pre

57、ferably drug product (or placebo) & known impurities at a minimum of three concentration level (each in triplicate), covering the minimum range required from quantification limit (impurity reporting limit), specification limit & level above the specification limit (150%) and calculate the % recovery

58、.备注Note: 必须配制控制溶液（药物产品），减去各准确度溶液原本残留的杂质，计算其%回收率。Control solution (drug product) must be prepared to subtract any residual impurity level to calculate the % recovery.各个水平的各杂质溶液的%回收率应该在80%-120%范围内。% Recovery of individual impurity at each level should be within 80 % to 120 %.精密度Precision方法精密度（重复性）Meth

59、od Precision (Repeatability)根据规定步骤，分别配制样品溶液6份（N=6），加入目标测试浓度100%水平的已知杂质（已有的）。Prepare sample solution six times (N=6) individually as per the defined procedure and spike the (available) known impurities at 100% level of the target test concentration. 确定6份不同样品中，所有单个杂质和总杂的%RSD。Determine the % RSD for al

60、l individual impurities & total impurities from six different samples.备注Note: 准确度实验中，100%回收率溶液可用于确定方法的精密度（重复性）。100% recovery solution (under accuracy experiment) can be used to determine the method precision (repeatability).单个杂质和总杂的%RSD 10% RSD (N=6) 10%, for both individual(s) & total impurities中间精