药品和生物制品分析方法验证和质量量度

GMP最前沿 医药国际注册与 GMP 俱乐部 俱乐部 QQ 群：292669664 GMP 最前沿 第九期分析方法验证和质?度? 编辑 by 付静 校验和审核 by 付静、 赵谦 评论：赵谦 本期介绍和讨论的两个 FDA 指南?别涉及到?析方法验证和质?度?的话题?析方法验证本身 不是什?新的概念，之前 FDA 出过不少相关的指南，ICH Q2 ?是?个话题?个新的版本区?了药典 方法和内控方法的一致性比较，融合了方法转移验证，并强调了在产品生命周期内的持续变更的要求? 笔者不认为?有任何新的或是更加明确的解释和阐述?一定要?强调的话，?只能?强调大家注 意?方法要适合其用途?个概念?因为引入了风险评估的概念，更加实际的操作可能体?在方法?验 证?一致性评价?转移验证等等实际操作中，验证的哪些项目可以做，哪些项目可以不做，做到什?程 度，可能都可以做到实际的?有据可查?，更加客观和科学了?对于引入的?方法性能?的概念，以及 对方法性能的周期性评价，实际?可以企业自行完善药典方法，注?方法的渠道和机会?方法性能? 和?方法验证风险评估?的结合体，可以给企业实际操作过程提供更大的灵活性，?点是所有实验室? 理需要结合自己的产品去深思熟虑，并想办法加以利用的? 质?度?的话题一?提出， 很多企业?惯性的就是反感， 总是觉得 FDA 是在加强监管? 但是就 FDA 目前强制要求提供的数据来看，?是非常初?的，围绕不合格率做的文章?就?个不合格率本身来说， 其实很难实际说明工厂的 GMP 状态?对于我们常看到中国和印度的企业来说，他们会采用反复测试? 试针? 不体?返工等等方式来做到不合格率为零， 如果考虑到?样的操作过程本身， 自然是不符合 CGMP 要求的，但是就最后的结论来说，一个企业愿意多花了更多的资源去保证自己出厂的产品最终质?是合 格的，那?是否?达到了 FDA 监管的目的呢？?本身就有方法学?的悖论存在?我们当然希望我们的 任何工厂都能在过程?完全符合 GMP 然后产品的不合格率?非常低?但是那些过程控制很差的企业， 为了保证不合格率低，违反 GMP 的原则，采用自己能采用的一?方法去降低不合格率，?样的操作本 身是否因为其目的的合理性可以被原谅呢？FDA 如果不原谅，那? FDA 收集质?度?的时候首先考虑 ?个指标又有什?意?呢？非常多的中国企业?任何行业?在做 ISO 的时候，在给大老板汇报自己的质 ?状态时都喜?说?100%合格?但是，他们都忽略了?个 100%合格的背景?前提和实?过程，?或 许是当?中国实体?济真的实?产业升级?需要加强和用力的地方，但是又有多少企业的老板真的认 识到?点呢？ 我们不希望FDA费尽心思收集的数据实际只增加了企业的负担， 而并不能真实的反馈企业的状态? 寄希望于简单的几个参数就可以反馈一个企业质?状态的想法本身是理想和完美的，但是可操作性到? 有多少？就我个人来说， 我更加相信过程中的参数， 实际简单的工艺参数的 CPk 就可以反?各个?节的 质?控制水平了，结合一定比例的权?因子，或许就能更加客观科学的反?企业的实际质?状态了?所 以在质?度?个事情?，或许 FDA 可以做的更加简单点?不过如果真的如果 FDA 简单的要求企业? 报所有关键质?参数的 CPk,那?多少中国企业会因?吐血？读者能自己回家算?，然后告诉我们吗？ 赵赵谦谦 GMP 最前沿 1 Analytical Procedures and Methods Validation for Drugs and Biologics 药药品品和和生生物物制制品品分分析析方方法法验验证证 Guidance for Industry1 ?业业指指南南 This guidance represents the current thinking of the Food and Drug Administration (FDA or Agency) on this topic. It does not create any rights for any person and is not binding on FDA or the public. You can use an alternative approach if it satisfies the requirements of the applicable statutes and regulations. To discuss an alternative approach, contact the FDA staff responsible for this guidance as listed on the title page. 本指南代表了 FDA ?本?题的?前想法?它并?赋?任何人以任何?利，也并? FDA 或? 形?强制效力?如果?方法?以满足?用的法规要求，你?使用?方法来?代?要?论?代性方 法，请联系列于封面的负责本指南的 FDA 职员? I. INTRODUCTION 概概述述 This guidance supersedes the draft of the same name that published on February 19, 2014 (79 FR 16 9467) and replaces the 2000 draft guidance for industry on Analytical Procedures and Methods Validation23and the 1987 Guidelines for Submitting Samples and Analytical Data for Methods Validation. It provides recommendations on how you, the applicant, can submit analytical procedures4and methods validation5data to support the documentation of the identity, strength, quality, purity, and potency of drug substances and drug products6.It will help you assemble information and present data to support your analytical methodologies. The recommendations apply to drug substances and drug products covered in new drug applications (NDAs), abbreviated new drug applications (ANDAs), biologics license applications (BLAs), and supplements to these applications. The principles in this guidance also apply to drug substances and drug products covered in Type II drug master files (DMFs). 本指南?代 2014 ? 2 ? 19 ?的?草案 ?79FR169467? ， ?代 2000 ?的?业指南 ?分析方法验证? 和 1987 ?的?递交?品和分析方法验证数据指南?指南?申?人如何提交分析方法验证数据来支持原料药和制剂鉴 别?剂?纯度和效?文?提供了建?指南?申?者如何收集资料，呈?数据来支持你的分析方法提 供了帮?些建?用于新药申?NDA?简略新药申?ANDA?生物药品许?申?BLA?以? 些申?的补充资料中的原料药和制剂?本指南中的原则也?用于?类药物?文?DMF?包括的原料药和制 剂? This guidance complements the International Conference on Harmonisation (ICH) guidance Q2(R1) Validation of Analytical Procedures: Text and Methodology (Q2(R1) for developing and validating analytical methods. 1This guidance has been prepared by the Office of Pharmaceutical Quality in the Center for Drug Evaluation and Research (CDER) and the Center for Biologics Evaluation and Research (CBER) at the Food and Drug Administration. 2Sample submission is described in section IX, FDA Methods Verification. 3We update guidances periodically. To make sure you have the most recent version of a guidance, check the FDA Drugs guidance Web page at /Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htm. 4Analytical procedure is interchangeable with a method or test procedure. 5Compendial methods are verified rather than validated as described in section VI, C. 6The terms drug substance and drug product are used in this guidance to refer to both human drugs and biologics. 2 GMP 最前沿 本指南补充 ICH 的指南 Q2(R1)分析方法验证?文和方法学?Q2(R1)?，?指南用于分析方法的开发和验证? This guidance does not address investigational new drug application (IND) methods validation, but sponsors preparing INDs should consider the recommendations in this guidance. For INDs, sufficient information is required at each phase of an investigation to ensure proper identity, quality, purity, strength, and/or potency. The amount of information on analytical procedures and methods suitability will vary with the phase of the investigation7.For general guidance on analytical procedures and methods validation information to be submitted for phase one studies, sponsors should refer to the FDA guidance for industry on Content and Format of Investigational New Drug Applications (INDs) for Phase 1 Studies of Drugs, Including Well-Characterized, Therapeutic, Biotechnology-Derived Products. General considerations for analytical procedures and methods validation before conduct of phase two and three studies are discussed in the FDA guidances for industry on INDs for Phase 2 and 3 Studies of Drugs, Including Specified Therapeutic Biotechnology-Derived Products (February 1999) and IND Meetings for Human Drugs and Biologics, Chemistry, Manufacturing, and Controls Information. 本指南并?论 IND 申?中的方法验证，但申请人在准备 IND 时?考虑本指南中的建?于 IND，要求一个 临床的各阶段都?充分的资料来确保?的鉴别?纯度?剂?和/或效?分析方法?用性的资料数?会 因临床阶段?而?一般来说，在一期临床提交的分析方法验证资料，需要参考 FDA ?业指南?药品一期 临床研究 IND 申?资料内容和格式，包括特性明确?治疗用生物?术?生?品?在?期和?期临床研究之前 的分析方法验证的一般考虑要点在 FDA ?业指南?药品的?期和?期临床研究 IND 申?资料?和?IND 申?评 ?会?人用药和生物制品?CMC 资料的要求?中?论? This guidance does not address specific method validation recommendations for biological and immunochemical assays for characterization and quality control of many drug substances and drug products. For example, some bioassays are based on animal challenge models, and immunogenicity assessments or other immunoassays have unique features that should be considered during development and validation. 本指南并没?给出原料药和药品的?制和生物和免疫化学含?表?测定特定的方法验证建? 例如， ?些生 物测试是?于?物挑战模式， 免疫?因评估或?它?独特性?的免疫分析， ?些分析在方法开发和验证时需要 考虑? Analytical methods required during product and process development activities are discussed in FDA guidance for industry on Process Validation: General Principles and Practices. 在药品和?艺研发期间?需的分析方法?在 FDA ?业指南?艺验证?一般原则和规范?中?了?论? In addition, a risk-based approach on the need for revalidation of existing analytical methods may need to be considered when the manufacturing process changes during the products life cycle. For questions on appropriate validation approaches for analytical procedures or submission of information not addressed in this guidance, you should consult with the appropriate FDA quality assessment staff. ?外， 在?品的生?周期中?生?艺?更时， ?能需要采用?于风险的方法考虑是?需要?分析方法? ?验证?于本指南中未?论的关于分析方法?的验证方式或资料提交的问题，?向?的 FDA ?评估 人员咨询? If you choose a different approach than those recommended in this guidance, we encourage you to discuss the matter with the appropriate FDA quality assessment staff before you submit your application. 如果你选择了一个?于本指南中的方法，?们鼓励你在提交申?资料前?的 FDA ?评估人员就? ?论? In general, FDAs guidance documents do not establish legally enforceable responsibilities. Instead, guidances describe the Agencys current thinking on a topic and should be viewed only as recommendations, unless specific regulatory or 7See 21 CFR 312.23(a)(7). GMP 最前沿 3 statutory requirements are cited. The use of the word should in Agency guidances means that something is suggested or recommended, but not required. 一般来说，FDA 的指南文?没?法律强制性?指南只是?述了?局目前?某个?题的考虑，除了?中引用的特 定的法规或法定要求外，本指南?仅作?是建?来看待?在官方指南中用词?表示建?或?荐某?，但并 ?是强制要求? 评评论论?总的来说，?个部分指明的是本法规?用的?象，除了 IND 申?方法?时?能调整的原因?生物? 术?品中方法开发和验证的特?的需求之外，?原料药?制剂?品?生物?术?品?熟的方法验证均?在? 个范围内?用?于风险的方法考虑，特别是你的注?方法和法定方法之间的?别和评估，法定方法?断更新和 ?化情况?，你的注?方法的?用性，参数设定的合理性，以?在什?情况?需要?据什?风险判断去做?态的 验证和?等等，?些都是本指南新?的?域和思考? II. BACKGROUND 背背景景 Each NDA and ANDA must include the analytical procedures necessary to ensure the identity, strength, quality, purity, and potency of the drug substance and drug product8.Each BLA must include a full description of the manufacturing process, including analytical procedures that demonstrate the manufactured product meets prescribed standards of identity, quality, safety, purity, and potency9.Data must be available to establish that the analytical procedures used in testing meet proper standards of accuracy, sensitivity, specificity, and reproducibility and are suitable for their intended purpose10. ?个 NDA 和 ANDA 都必?包括用以确保原料药和制剂鉴别?剂?纯度和效?的分析方法?个 BLA 必?包括生?艺的全面?述，包括用以证明?生?品符合?述鉴别?案例?纯度和效?标准的分析方 法?必?数据证明用于测试的分析方法符合?的准确度?灵敏度?属性和?复性标准，并?合于?既定用 途? Analytical procedures verification or validation data should be submitted in the corresponding sections of the application in the ICH M2 eCTD: Electronic Common Technical Document Specification11. 分析方法确认或验证数据?在 ICH M2 eCTD?电子通用?术文?规范?申?资料中相?的部分提交? When an analytical procedure is approved/licensed as part of the NDA, ANDA, or BLA, it becomes the FDA-approved analytical procedure for the approved product. This analytical procedure may originate from FDA recognized sources (e.g., a compendial procedure from the United States Pharmacopeia/National Formulary (USP/NF) or a validated procedure you submitted that was determined to be acceptable by FDA. To apply an analytical method to a different drug product, appropriate validation or verification studies for compendial procedures with the matrix of the new product should be considered. 如果分析方法作? NDA?ANDA 或 BLA 的一部分被批准，则? FDA ?批准?品的分析方法的批准? 分析方法?以是来源于 FDA 认?的来源?例如，USP/NF 药?方法?或一个你?提交的?过验证的方法，?被 FDA 认?是?以接?的?在?药品中?用一个分析方法时，要考虑?入?药?方法?的验证或确认 研究? 8See 21 CFR 314.50(d)(1) and 314.94(a)(9)(i). 9See 21 CFR 601.2(a) and 601.2(c). 10See 21 CFR 211.165(e) and 211.194(a)(2). 11Sections as applicable in Module 3: 3.2.S and 3.2.P. 4 GMP 最前沿 评评论论?背景确认了方法验证的?本内容，确认了在递交文?中必?涵盖方法验证的规定?在 FDA 眼?，USP 的 方法和你自?内?验证的方法?实是地?一?的? 如果你?以?数据证明， 那?一定情况?你的内?验证方法? 能? USP 的方法会更?获得 FDA 的认?和支持?个地?的?，一直困扰了?少?要做?内?场的企业在做 ?化的时候?确的?待 USP 的方法?时，也因?个原因，很多企业在明明知道 CP ?合自?的?品的时 候，?了简化也好，?了?得罪?威也好，都没?站出来指明 CP 的缺陷，中?的药?员会也没?一个?效的 通道去允许企业?极的参?到?方法的?合理?论中去? ?是值得中?药?员会和?家药?局思考和改良的 方面，依靠?十个人的被?修?千个标准，?多的资金和资源都?如化整?零，?员实?生?品的企业去完 善和建立?术壁垒?用?场手段解决了革新?力的问题?，或许才能?本?解决标准落?以?管?力的 问题? III. ANALYTICAL METHODS DEVELOPMENT 分分析析方方法法开开发发 An analytical procedure is developed to test a defined characteristic of the drug substance or drug product against established acceptance criteria for that characteristic. Early in the development of a new analytical procedure, the choice of analytical instrumentation and methodology should be selected based on the intended purpose and scope of the analytical method. Parameters that may be evaluated during method development are specificity, linearity, limits of detection (LOD) and limits of quantitation (LOQ), range, accuracy, and precision. 开发分析方法是?了测试原料药或药品的指定属性， 以确认?是?符合?建立的?属性?接?标准? 在新分析方 法的开发?期， ?据?用目的和范围来选择?用的分析仪器和方法? 在方法开发中需要?评估的参数? 属性?线性?检测限?LOD?和定?限?LOQ?范围?准确度和精密度? During early stages of method development, the robustness of methods should be evaluated because this characteristic can help you decide which method you will submit for approval. Analytical procedures in the early stages of development are initially developed based on a combination of mechanistic understanding of the basic methodology and prior experience. Experimental data from early procedures can be used to guide further development. You should submit development data within the method validation section if they support the validation of the method. 在方法开发?期，?方法的耐用性?评估，因?属性?以帮?你决定提交哪个方法去批准?分析方法?期 开发是?于?础方法学的了解和之前的?验来建立的? ?期程序的实验数据?以用于指?一?的开发? 如果 ?些开发数据支持方法验证的话，申请者?在方法验证部分提交?数据? 评评论论?提出了在方法开发的?期，首要考虑的问题居然涵盖耐耐用用性性！实?在?们的方法开发中，更多更? 侧?的是?属性?也就是选择性?的问题，?真的很少考虑耐用性的问题?耐用性指的是微小的方法参数的调整 一?以获得很好的?性！?如体系的 pH?如流?如柱温?如?度的调整等等?想?个要求是 合理的，如果你的方法选择性很好，但是非常?耐用，pH 一点点的波?就会?你的峰分?开了，?些信?如 果?的确认，?能?续你确认的?谓方法，也就要?新开发了?个提醒真的很?值！ ?外开发阶段的数据很少?以用于方法验证的递交数据， 但是如果?以用于支持某些结论， 那就?以递交用来支 持?换?话说，?期开发阶段用到的实验条?，如果?以用于证明?是耐用的，那?在?期方法验证阶段是?需 要?新做了，直接引用开发阶段的耐用性数据即?！?个显然也是合理的?就看企业的策略和管理方式了? To fully understand the effect of changes in method parameters on an analytical procedure, you should adopt a systematic approach for a method robustness study (e.g., a design of experiments with method parameters). You should begin with an initial risk assessment and follow with multivariate experiments. Such approaches allow you to understand factorial parameter effects on method performance. Evaluation of a methods performance may include analyses of samples obtained from various stages of the manufacturing process from in-process to the finished product. Knowledge gained during these studies on the sources of method variation can help you assess the method performance. GMP 最前沿 5 ?了全面了解分析方法参数?更的影响，?采用一个系统的方法?方法耐用性研究?例如，设?一个方法参 数实验?首?采用风险评估，然?多?实验?的方法能帮?了解参数因子?方法性能的影响? ?方法性能评估?以通过分析来自生?艺中包括中?到?品?阶段的?品? ?些方法?化来源的研究中获 得的知识?以帮?评估方法的性能? 评评论论? ?实?是在操作层面指?企业如何去设?合理的方法耐用性， 多因子?交实验方法的设?以很好 的满足?个要求，?也是显而易?的?明显?势的地方，就是?方法本身的性能在各方面?多个潜在影响因子的 各方面?都?全面的理解和了解? IV. CONTENT OF ANALYTICAL PROCEDURES 分分析析方方法法的的内内容容 You should describe analytical procedures in sufficient detail to allow a competent analyst to reproduce the necessary conditions and obtain results within the proposed acceptance criteria. You should also describe aspects of the analytical procedures that require special attention. An analytical procedure may be referenced from FDA-recognized sources (e.g., USP/NF, Association of Analytical Communities (AOAC) International)12, if the referenced analytical procedure is not modified beyond what is allowed in the published method. You should provide in detail procedures from other published sources. The following is a list of essential information you should include for an analytical procedure: ?于分析方法?述需要足够?，使得?资?的化验员?以?必要的条?，获得?拟?接?标准内的结果?分 析方法中需要特别注意的方面也?述? 如果被引用的分析方法仅在?允许的范围内?方法?修 改，那?以直接引自 FDA 认?的来源?例如 USP/NF，?分析共?体协会?AOAC?的分析方法?需要提 供?它出版来源的?方法?以?是一个分析方法中?包括的?本信?清单? A. Principle/Scope 原原理理/范范围围 A description of the basic principles of the analytical test/technology (i.e., separation, detection); target analyte(s) and sample(s) type (e.g., drug substance, drug product, impurities or compounds in biological fluids). 分析测试/?术?如?分离?检测?本原理的?述?目标分析物和?品类型?例如，原料药?制剂?或生 物流体中的化合物? B. Apparatus/Equipment 仪仪器器/设设备备 All required qualified equipment and components (e.g., instrument type, detector, column type, dimensions, and alternative column, filter type). ?需要的确认过的仪器和?例如，仪器类型?检测器?柱子类型?尺?和?代的柱子?过滤器类型? C. Operating Parameters ?参参数数 Qualified optimal settings and ranges (include allowed adjustments supported by compendial sources or development and/or validation studies) critical to the analysis (e.g., flow rate, components temperatures, run time, detector settings, gradient, head space sampler). A drawing with experimental configuration and integration parameters may be used, as applicable. 确认过的最佳的设置和范围?包括药?来源或研发和/或验证研究的允许调整?，?于分析过程非常关键的设置 ?例如，流?部?温度?时间?检测器设置?度?顶空?器?时，?以?录实?的方法配置和 ?分参数的? D. Reagents/Standards 试试剂剂/标标准准 The following should be listed where applicable: 12See 21 CFR 211.194(a)(2). 6 GMP 最前沿 ?时?列出以?内容 Description of reagent or standard 试剂或标准的?述 Grade of chemical (e.g., USP/NF, American Chemical Society, High Performance or Pressure Liquid Chromatography, or Gas Chromatography and preservative-free) 化学品的?别?例如，USP/NF，美?化学协会，HPLC 色谱?，或 GC 色谱?或无防腐剂的? Source (e.g., USP reference standard, qualified in-house reference material, WHO International Standard/Reference Material, CBER standard) 来源?例如，USP 标准品，确认过的内部?照物?，WHO ?标准/?照物?，CBER 标准? Purity (for pure chemicals only), State (e.g., dried, undried), and concentration 纯度?只?纯的化学品需要?状态?例如，?品，未?燥品?和浓度 Potencies (where required by CFR, USP) 效?CFR，USP ?要求? Storage conditions ?条? Directions for safe use (as per current Safety Data Sheet) ?全使用指示?以? SDS ?准? Validated or documented shelf life 验证过的或记录的?效期 New batches of biological reagents, such as monoclonal antibodies, polyclonal antisera, or cells, may need extensive qualification procedures included as part of the analytical procedure. 生物试剂的新批?，例如，单克隆抗体?多克隆抗原?或?胞，?能需要包括?一?确认程序，作?分析方法的 一部分? E. Sample Preparation ?品品制制备备 Procedures (e.g., extraction method, dilution or concentration, desalting procedures and mixing by sonication, shaking or sonication time) for the preparations for individual sample tests. A single preparation for qualitative and replicate preparations for quantitative tests with appropriate units of concentrations for working solutions (e.g., g/ml or mg/ml) and information on stability of solutions and storage conditions. 各供试?品的制备程序?例如，提?方法?稀释或浓缩?除?和超声混合?震摇或超声时间?供试?定性单? 配制方法，定?测试?品配制方法，?作溶液?的浓度单?例如 g/ml 或 mg/ml?，以?溶液的稳定性 和?条?的信? 评评论论?分析方法需要指明溶液的稳定性?点一般是在做方法验证的时候确定的内容?在明确提出要?方法 ?述中去了? F. Standards Control Solution Preparation 标标准准?制制溶溶液液制制备备 Procedures for the preparation and use of all standard and control solutions with appropriate units of concentration and information on stability of standards and storage conditions, including calibration standards, internal standards, system suitability standards, etc. GMP 最前沿 7 ?标准和?制溶液的制备和使用程序?的浓度单?，?标准溶液稳定性信?和?条?，包括校?标 准溶液?内部标准溶液?系统?用性标准溶液等? G. Procedure 检检验验程程序序 A step-by-step description of the method (e.g., equilibration times, and scan/injection sequence with blanks, placeboes, samples, controls, sensitivity solution (for impurity method) and standards to maintain validity of the syst