FDA行业指南：分析方法验证（中英文）

http://zhuyujiao1972.blog.163.com/。
. D4 V  l/ e- r, a/ Q) ?$ ^

Analytical Procedures and Methods Validation for Drugs and Biologics

药物和生物制品分析方法验证

Guidance for Industry[1]

行业指南

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对本专题的当前想法。它并不赋予任何人以任何权利，也并不对FDA或公众形成强制效力。如果有方法可以满足适用的法规要求，你可使用该方法来替代。要讨论替代性方法，请联系列于封面的负责本指南的FDA职员。

I. INTRODUCTION 概述

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supersedes the draft of the same name that published on February 19, 2014 (79 FR 16 9467) and replaces the 2000 draft guidance for industry on Analytical Procedures and Methods Validation[2][3]and the 1987 Guidelines for Submitting Samples and Analytical Data for Methods Validation. It provides recommendations on how you, the applicant, can submit analytical procedures[4]and methods validation[5]data to support the documentation of the identity, strength, quality, purity, and potency of drug substances and drug products[6].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).

本指南取代2014年2月19日公布的同名草案（79FR169467），替代2000年的行业指南“分析方法验证”和1987年的“提交方法验证的样品和分析数据指南”。它指导你、申报人如何提交分析方法验证数据来支持原料药和制剂鉴别、剂量、质量、纯度和效价文件。它会帮助你组织资料，呈现数据来支持你的分析方法学。建议适用于新药申报（NDA）、简略新药申报（ANDA）、生物药品许可申报（BLA）以及对这些申报的补充资料中的原料药和制剂。本指南中的原则也适用于二类药物主文件（DMF）所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.

本指南补充ICH的指南Q2(R1)分析方法验证：正文和方法学（Q2(R1)），该指南用于分析方法的研发和验证。

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 investigation[7].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.

本指南并不讨论IND申报中的方法验证，但申请人在准备IND时应考虑本指南中的建议。对于IND，要求一个临床的各阶段都有充分的资料来确保适当的鉴别、质量、纯度、剂量和/或效价。分析方法适用性的资料数量会因临床阶段不同而不同。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行业指南“药品一期临床研究IND申报资料内容和格式，包括特性明确、治疗用生物技术衍生产品”。在二期和三期临床研究之前的分析方法验证的一般考虑要点在FDA行业指南“药品的二期和三期临床研究IND”和“人用药和生物制品、CMC资料IND会议”中有讨论。

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.

本指南并不讨论许多原料药和药品的质量控制和生物和免疫化学测定的定性中特定的方法验证建议。例如，有些生物含量是基于动物挑战模式，免疫基因评估或其它免疫测试具有独特的性质，在研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.

在药品和工艺研发期间所需的分析方法已在FDA行业指南“工艺验证：一般原则和规范”中进行了讨论。

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 product’s 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.

此外，在产品的生命周期中当生产工艺变更时，可能需要采用基于风险的方法考虑是否需要对现有分析方法进行再验证。对于本指南中未讨论的关于分析方法适当的验证方法或资料提交的问题，你应当向适当的FDA质量评估人员咨询。

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.

如果你选择了一个不同于本指南中的方法，我们鼓励你在提交申报资料前与适当的FDA质量评估人员就此事进行讨论。

In general, FDA’s guidance documents do not establish legally enforceable responsibilities. Instead, guidances describe the Agency’s current thinking on a topic and should be viewed only as recommendations, unless specific regulatory or 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的指南文件没有法律强制性。指南只是描述了当局目前对某个专题的考虑，除了其中引用的特定的法规或法律条款要求外，应当仅作为是建议来看待。在官方指南中用词“应”表示建议或推荐某事，但并不是强制要求。

II. BACKGROUND 背景

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 product[8].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 potency[9].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 purpose[10].

每个NDA和ANDA都必须包括用以确保原料药和制剂鉴别、剂量、质量、纯度和效价的分析方法。每个BLA必须包括生产工艺的全面描述，包括用以证明所生产产品符合所述鉴别、质量、案例、纯度和效价标准的分析方法。必须有数据证明用于测试的分析方法符合适当的准确度、灵敏度、专属性和重复性标准，并适合于其既定用途。

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 Specification[11].

分析方法确认或验证数据应在ICH M2 eCTD“电子通用技术文件规范”申报资料中相应的部分提交。

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.

如果分析方法作为NDA、ANDA或BLA的一部分被批准，则其成为FDA批准产品的批准的分析方法。该分析方法可以是来源于FDA认可的来源（例如，USP/NF药典方法）或一个你所提交的经过验证的方法，而被FDA认为是可以接受的。在不同药品中应用一个分析方法时，要考虑加入新药的基底后对药典方法进行适当的验证或确认研究。

III. ANALYTICAL METHODS DEVELOPMENT 分析方法研发

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.

研发分析方法是为了测试原料药或药品的指定属性，以确认其是否符合已建立的该属性可接受标准。在新分析方法的研发早期，应根据其应用目的和范围来选择所用的分析仪器和方法。在方法研发中需要进行评估的参数为专属性、线性、检测限（LOD）和定量限（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.

在方法研发早期，应对方法的耐用性进行评估，因为该属性可以帮助你决定提交哪个方法去批准。研发早期的分析方法最早是基于对基础方法学的了解和之前的经验来建立的。早期程序的实验数据可以用于指导进一步的研发。如果这些研发数据支持方法的验证的话，你应该在方法验证部分提交研发数据。

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 method’s 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.

为了全面了解分析方法参数变更的影响，你应该采用一个系统的方法进行方法耐用性研究（例如，设计一个方法参数实验）。开始你应采用风险评估，然后进行多变量实验。这样的方法能让你了解参数因子对方法性能的影响。对方法性能评估可以包括分析来自生产工艺中从中控到成品不同阶段的样品。从这些方法变化来源的研究中获得的知识可以帮助你评估方法的性能。

IV. CONTENT OF ANALYTICAL PROCEDURES 分析方法的内容

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:

你应该将分析方法叙述的足够详细，使得有资质的化验员可以重现必要的条件，获得所拟可接受标准内的结果。你还应该描述分析方法所需的注意事项。分析方法可以引自FDA认可的来源（例如USP/NF，国际分析共同体协会（AOAC）），只要被引用的分析方法未修订至超出所公布方法所允许的范围。你应该提供其它出版来源的详细方法。以下是一个分析方法中应该包括的基本信息清单：

A. Principle/Scope 原理/范围

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/技术（即分离、检测）基本原因的描述；目标分析物和样品类型（例如，原料药、制剂、杂质或生物流体中的化合物）。

B. Apparatus/Equipment 仪器/设备

All required qualified equipment and components (e.g., instrument type, detector, column type, dimensions, and alternative column, filter type).

所有需要的确认过的仪器和组件（例如，仪器类型、检测器、柱子类型、尺寸和可替代的柱子、过滤器类型）。

C. Operating Parameters 运行参数

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.

确认过的优化的设置和范围（包括来自药典或研发和/或验证研究的允许调整），对于分析过程非常关键（例如，流速、部件温度、运行时间、检测器设置、梯度、顶空进样器）。适当时，可以使用经验参数设置和积分参数的样图。

D. Reagents/Standards 试剂/标准

The following should be listed where applicable:

适当时应列出以下内容

?   Description of reagent or standard

?   试剂或标准的描述

?   Grade of chemical (e.g., USP/NF, American Chemical Society, High Performance or Pressure Liquid Chromatography, or Gas Chromatography and preservative-free)

?   化学品的级别（例如，USP/NF，美国化学协会，HPLC色谱级，或GC色谱级或无防腐剂的）

?   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，内部确认的对照物质，WHO国际标准/对照物质，CBER标准）

?   Purity (for pure chemicals only), State (e.g., dried, undried), and concentration

?   纯度（只有纯的化学品需要）、状态（例如，干品，未干燥品）和浓度

?   Potencies (where required by CFR, USP)

?   效价（CFR，USP所要求）

?   Storage conditions

?   存贮条件

?   Directions for safe use (as per current Safety Data Sheet)

?   安全使用指示（以现行SDS为准）

?   Validated or documented shelf life

?   验证过的或记录的货架期

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.

生物试剂的新批号，例如，单克隆抗体、多克隆抗原、或细胞，可能需要包括进一步确认程序，作为分析方法的一部分。

E. Sample Preparation 样品制备

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., μg/ml or mg/ml) and information on stability of solutions and storage conditions.

各供试样品的制备程序（例如，提取方法、稀释或浓缩、除盐和超声混合、震摇或超声时间）。供试样定性单样配制方法，定量测试平行样品配制方法，工作溶液适当的浓度单位（例如μg/ml或mg/ml），以及溶液的稳定性和存贮条件的信息。

F. Standards Control Solution Preparation 标准控制溶液制备

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.

所有标准和控制溶液的制备和使用程序。具有适当的浓度单位，有标准稳定性信息和存贮条件，包括校正标准、内部标准、系统适用性标准等。

G. Procedure 检验程序

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 system suitability during the span of analysis) and allowable operating ranges and adjustments if applicable.

对方法的逐步描述（例如，平衡时间，扫描/进针序列，空白，基底样，样品，控制样，敏感溶液（杂质方法）和在分析过程中维持系统适用有效性的标准样）以及允许运行范围和适当时的调整。

H. System Suitability 系统适用性

Confirmatory test(s) procedures and parameters to ensure that the system (equipment, electronics, and analytical operations and controls to be analyzed) will function correctly as an integrated system at the time of use. The system suitability acceptance criteria applied to standards controls and samples, such as peak tailing, precision and resolution acceptance criteria, may be required as applicable. For system suitability of chromatographic systems, refer to the FDA guidance for industry on Validation of Chromatographic Methods and USP General Chapter <621>Chromatography.

对测试程序和参数进行确证以保证系统（仪器、电子和分析操作和要分析的控制点）能在使用时作为一个整体正确运行。适用于对照控制和样品的系统适用性的可接受标准，如拖尾因子、精密度和分辨率可接受标准，在适当时可以进行要求。色谱系统的系统适用性，参见FDA行业指南“色谱方法的验证”和USP通论<621>色谱。

I. Calculations 计算

The integration method and representative calculation formulas for data analysis (standards, controls, samples) for tests based on label claim and specification (e.g., assay, specified and unspecified impurities and relative response factors). This includes a description of any mathematical transformations or formulas used in data analysis, along with a scientific justification for any correction factors used.

根据标识声明和质量标准（例如，含量、特定和非特定杂质和相对响应因子）进行测试所得的数据分析（标准、控制、样品）中所用的积分方法和代表性计算公式。其中应包括数据分析中所使用的所有数学变换或公式的描述，以及使用的所有修正因子的科学论证。

J. Data Reporting数据的报告

A presentation of numeric data that is consistent with instrumental capabilities and acceptance criteria. The method should indicate what format to use to report results (e.g., percentage label claim, weight/weight, and weight/volume) with the specific number of significant figures needed. The American Society for Testing and Materials (ASTM) E29 standard describes a standard practice for using significant digits in test data to determine conformance with specifications. For chromatographic methods, you should include retention times (RTs) for identification with reference standard comparison basis, relative retention times (RRTs) (known and unknown impurities) acceptable ranges and sample results reporting criteria.

与仪器的能力和可接受标准相一致的数字式数据的呈现方式。方法中应指明要采用何种格式来报告结果（例如，标识声明的百分比，重量/重量，重量/体积），并指定所需报告的有效位数。美国材料试验协会（ASTM）E29标准中描述了在测试数据中使用有效位数来决定与质量标准符合性的标准规范。如果是色谱方法，你应该包括保留时间（RT）用于与对照品比较，相对保留时间（RRT）（已知和未知杂质）可接受范围和样品结果报告标准。

V. REFERENCE STANDARDS AND MATERIALS 对照标准和物质

Primary and secondary reference standards and materials are defined and discussed in the following ICH guidances: Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products, and Q7 Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients. For all standards, you should ensure the suitability for use. You should strictly follow storage and usage conditions and handling instructions for reference standards to avoid modifications and contaminations, which could result in additional impurities and inaccurate analysis. You should include information supporting any reference standards and materials that you intend to use in the application. Information supporting reference standards and materials should include qualification test reports and certificates of analysis (including stability protocols, reports, and relevant known impurity profile information) as applicable. For biological products under BLAs, qualification of subsequent reference standard lots should be included in annual reports.

基准对照品和第二对照品和物质在以下ICH指南中已有定义和讨论：Q6B“质量标准：生物技术/生物制品检验方法和可接受标准”和Q7“原料药GMP指南”。你要确保所有的标准品均适合于其用途。你要严格遵守对照品存贮和使用条件及处理指令，避免改动和污染，这可能会导致另外的杂质和分析不准确。你应该在申报资料中包括各对照品和物质适合于其用途的支持信息。适当时，支持对照品和物质的信息应包括确认试验报告和检验报告（包括稳定性试验方案、报告、相关已知杂质概况资料）。对于BLA下的生物制品，要在年报中包括之后的对照品批次的确认。

Reference standards can often be obtained from USP and may also be available through the European Pharmacopoeia, Japanese Pharmacopoeia, World Health Organization, or National Institute of Standards and Technology. Reference standards for a number of biological products are also available from CBER. For certain biological products marketed in the U.S., reference standards authorized by CBER must be used before the product can be released to the market[13].Reference materials from other sources should be characterized by procedures including routine and beyond routine release testing as described in ICH Q6B. You should consider orthogonal methods for reference material characterization. Additional testing could include attributes to determine the suitability of the reference material not necessarily captured by the drug substance or product release tests (e.g., more extensive structural identity and orthogonal techniques for potency, purity and impurities).

对照品通常可以从USP处获得，也可以通过EP、JP、WHO或国际标准技术委员会获得。大量的生物产品对照品也可以从CBER处获得。在美国上市的特定生物制品，CBER授权的对照品必须在产品放行上市前使用。从其它来源获得的对照物质应根据程序进行确证，包括常规测试和ICH Q6B里所述的超出常规放行测试的项目。你应考虑矩阵方法来确证对照品。附加测试可以包括确定对照物质适用性，这可能在原料药或药品放行测试中发现不了（例如，更全面的结构确证和效价、纯度和杂质矩阵技术）。

A new batch of reference standard material (official or in-house) should be qualified/calibrated against the current reference standard. For biological reference standards and materials, we recommend that you follow a two-tiered approach when qualifying new reference standards to prevent drift in the quality attributes. A two-tiered approach involves a comparison of each new reference standard with a primary reference standard so that it is linked to clinical trial material and the current manufacturing process.

新的对照品物质（官方的或自制的）要采用现行的对照品进行确认/校正。对于生物制品对照品和物质，我们建议你在确认新的对照品时采用双轨方法，以防止质量属性的漂移。双轨方法是将每个新的对照品与基准对照品进行比较，这样将其与临床试验物质和现行生产工艺相关联。

VI. ANALYTICAL METHOD VALIDATION 分析方法验证

A. Noncompendial Analytical Procedures 非药典分析方法

Analytical method validation is the process of demonstrating that an analytical procedure is suitable for its intended purpose. The methodology and objective of the analytical procedures should be clearly defined and understood before initiating validation studies. This understanding is obtained from scientifically-based method development and optimization studies. Validation data must be generated under a protocol approved by the sponsor following current good manufacturing practices with the description of methodology of each validation characteristic and predetermined and justified acceptance criteria, using qualified instrumentation[14].Protocols for both drug substance and product analytes or mixture of analytes in respective matrices should be developed and executed. You should include details of the validation studies and results with your application.

分析方法验证是证明一个分析方法适合于其既定用途的过程。方法学和分析方法的目的应在开始验证研究之前进行清楚的界定和了解。这种了解是来自于基于科学的方法研发和优化研究的。验证数据必须是从发起方按GMP规范批准的方案中产生，方案中要有每个验证项目的方法学描述，并确定和论述可接受标准，采用经过确认的仪器实施验证。应起草并实施原料药和制剂分析物或对应基质的分析混合物的验证方案。你应该在你的申报资料中包括验证研究及结果的详细内容。

B. Validation Characteristics 验证属性

Although not all of the validation characteristics are applicable for all types of tests, typical validation characteristics are:

尽管并不是所有的验证属性适用于所有类型的测试，但一般的验证属性包括：

?   Specificity 专属性

?   Linearity 线性

?   Accuracy 准确度

?   Precision (repeatability, intermediate precision, and reproducibility) 精密度（重复性、中间精密度和再现性）

?   Range 范围

?   Quantitation limit 定量限

?   Detection limit 检出限

ICH Q2(R1) is considered the primary reference for recommendations and definitions on validation characteristics for analytical procedures. The FDA guidance for industry on Validation of Chromatographic Methods is available as well.

ICH Q2(R1)被认为是是分析方法验证属性的建议和定义的基本参考资料。FDA行业指南“色谱方法验证”也可以找到。

If a procedure is a validated quantitative analytical procedure that can detect changes in a quality attribute(s) of the drug substance and drug product during storage, it is considered a stability-indicating test. To demonstrate specificity of a stability-indicating test, a combination of challenges should be performed. Some challenges include the use of samples spiked with target analytes and all known interferences; samples that have undergone various laboratory stress conditions; and actual product samples (produced by the final manufacturing process) that are either aged or have been stored under accelerated temperature and humidity conditions.

如果一个方法是经过验证的定量分析方法，可以检出原料药和制剂在存贮期间的质量属性的变化，则认为该方法是具有稳定性指示性的测试方法。为了证明一个稳定性指示检验方法的专属性，要进行一系列的挑战。一些挑战包括向样品中加入目标分析物和所有已知的干扰物、经过不同化验室强降解试验的样品、实际药品的样品（通过最终生产工艺生产出）经过长期放置或在加速温湿度条件下存贮。

As the holder of the NDA, ANDA, or BLA, you must: (1) submit the data used to establish that the analytical procedures used in testing meet proper standards of accuracy and reliability, and (2) notify the FDA about each change in each condition established in an approved application beyond the variations already provided for in the application, including changes to analytical procedures and other established controls[15].

作为NDA、ANDA或BLA的持有人，你必须（1）提交数据用于支持所用分析方法符合准确度和可靠性要求，（2）通知FDA在已批准的申报资料中所建立的每个条件超出申报资料中已批准变化范围的每个变更，包括对分析方法和其它已建立的控制的变更。

The submitted data should include the results from the robustness evaluation of the method, which is typically conducted during method development or as part of a planned validation study[16].

所提交的数据应包括方法耐用性评估结果，该项目一般是在方法研发时做的，或者作为验证研究计划的一部分。

C. Compendial Analytical Procedures 药典分析方法

The suitability of an analytical procedure (e.g., USP/NF, the Official Methods of Analysis of AOAC International, or other recognized standard references) should be verified under actual conditions of use[17].Information to demonstrate that USP/NF analytical procedures are suitable for the drug product or drug substance should be included in the submission and generated under a verification protocol.

一个分析方法（例如，USP/NF，AOAC国际的官方分析方法，或其它公认的标准对照）的适用性应在实际使用条件下经过确认。证明USP/NF分析方法适用于原料药或制剂的资料应包括在申报资料里，并且是从确认方案中产生的。

The verification protocol should include, but is not limited to: (1) compendial methodology to be verified with predetermined acceptance criteria, and (2) details of the methodology (e.g., suitability of reagent(s), equipment, component(s), chromatographic conditions, column, detector type(s), sensitivity of detector signal response, system suitability, sample preparation and stability). The procedure and extent of verification should dictate which validation characteristic tests should be included in the protocol (e.g., specificity, LOD, LOQ, precision, accuracy). Considerations that may influence what characteristic tests should be in the protocol may depend on situations such as whether specification limits are set tighter than compendial acceptance criteria, or RT or RRT profiles are changing in chromatographic methods because of the synthetic route of drug substance or differences in manufacturing process or matrix of drug product. Robustness studies of compendial assays do not need to be included, if methods are followed without deviations.

确认方案应包括，但不仅限于（1）要确认的药典方法，预定的可接受标准，（2）方法学的详细说明（例如，试剂、设备、配件、色谱条件、色谱柱、检测器类型、检测器信号响应灵敏度、系统适用性、样品制备和其稳定性）。该方法和确认程度应决定要包括在方案中的验证项目测试（例如，专属性、LOD、LOQ、精密度、准确度）。在决定方案中应包括哪些验证项目时的考虑取决于一些具体情况，如质量标准限度设定是否严于药典可接受标准，或RT或RRT概况由于原料药合成路线、生产工艺或制剂基底不同而在色谱方法中有所变化。如果完全按照药典方法实施，没有偏差的话，药典含量方法不需要进行耐用性研究。

VII. STATISTICAL ANALYSIS AND MODELS 统计学分析和模型

A. Statistics 统计学

Statistical analysis of validation data can be used to evaluate validation characteristics against predetermined acceptance criteria. All statistical procedures and parameters used in the analysis of the data should be based on sound principles and appropriate for the intended evaluation. Several statistical methods are useful for assessing validation characteristics, for example, an analysis of variance (ANOVA) to assess regression analysis R (correlation coefficient) and R squared (coefficient of determination) or linear regression to measure linearity. Many statistical methods used for assessing validation characteristics rely on population normality, and it is important to determine whether or not to reject this assumption. There are many techniques, such as histograms, normality tests, and probability plots that can be used to evaluate the observed distribution. It may be appropriate to transform the data to better fit the normal distribution or apply distribution-free (nonparametric) approaches when the observed data are not normally distributed. Appropriate literature or text should be consulted for information on statistical procedures to use when developing new test methods, evaluating existing test methods or evaluating measurement system performance, as well as other general information on the interpretation and treatment of analytical data[18].The data analysis should be assured either by using appropriately validated software or independent verification for correctness.

验证数据的统计学分析可以用于评估验证的属性是否符合预定的可接受标准。所有用于数据分析的统计学程序和参数均应是基于合理的原则，并适合于既定评估。有几个统计学方法用于评估验证属性颇为有用，例如，变量分析（ANOVA）用于评估相关性分析R（相关因子）和R平方（判定系数或拟合优度）或线性回归用于测量线性。许多用于评估验证属性的统计学方法依赖于样本的正态性，决定是否拒绝该假设很重要。有许多技术，如柱状图、正态分布和概率图，可以用于评估所观察到的分布情况。如果观察到的数据是非正态分布的，则将数据转换成为更为正态分布或应用非正态分布（无参数）方法会更为恰当。在研发新的分析方法、评估现有分析方法、或评估测量系统性能时，应参考适当的文献或文件来获取关于统计学程序的信息，以及关于分析数据诠释和处理的其它通用信息。数据分析应采用经过适当验证的软件，否则应单独确认其正确性。

B. Models 模型

Some analytical methods might use chemometric and/or multivariate models. When developing these models, the number of samples to provide adequate statistical power and range for model development and validation should be considered. Suitable software should be used for data analysis. Model parameters should be deliberately varied to test model robustness.

有些分析方法可能会使用化学计量学和/或多变量模型。如果研发的这些模型、样品数据可以提供足够的统计功效和范围用于建模，则应考虑进行验证。可以使用适当的软件进行数据分析。应该设计变化模型参数来测试模型的耐用性。

VIII. LIFE CYCLE MANAGEMENT OF ANALYTICAL PROCEDURES 分析方法的生命周期管理

Once an analytical procedure (including compendial methods) is successfully validated (or verified) and implemented, the procedure should be followed during the life cycle of the product to continually assure that it remains fit for its intended purpose. Trend analysis on method performance should be performed at regular intervals to evaluate the need to optimize the analytical procedure or to revalidate all or a part of the analytical procedure. If an analytical procedure can only meet the established system suitability requirements with repeated adjustments to the operating conditions stated in the analytical procedure, the analytical procedure should be reevaluated, revalidated, or amended, as appropriate.

分析方法（包括药典方法）被成功验证（或确认）和实施后，在其产品的生命周期中应遵守该方法，以持续保证方法保持适合其既定用途。应定期对方法表现进行趋势分析，评估是否需要对分析方法进行优化，或对全面或部分分析方法进行再验证。如果一个分析方法只能通过不断调整分析方法里载明的运行参数来符合所建立的系统适用性要求，则应对该分析方法进行再评估、再验证，适当时进行修正。

Over the life cycle of a product, new information and risk assessments (e.g., a better understanding of product CQAs or awareness of a new impurity) may warrant the development and validation of a new or alternative analytical method. New technologies may allow for greater understanding and/or confidence when ensuring product quality. Applicants should periodically evaluate the appropriateness of a product’s analytical methods and consider new or alternative methods.

在一个产品的整个生命周期中，新的资料和风险评估（例如，对产品CQA有更好的了解，或发现新的杂质）可能会保证一个新的或替代的分析方法的研发和验证。新技术可能会带来产品质量保证方面更多的了解和/或可信度。申报者应定期评估产品分析方法的适当性，考虑新的或可替代的方法。

In anticipation of life cycle changes in analytics, an appropriate number of retention samples should be maintained to allow for comparative studies. The number should be based on scientific principles and an assessment of risk. For complex products that are sensitive to manufacturing changes, reserve samples can be an important tool to make these comparisons.

预计在生命周期中会对分析方法进行变更，因此要保留适当数据量留样进行对比研究。样品数量应基于科学原理，以及风险评估。对生产工艺较为敏感的复杂产品，其留样可能是做对比研究的重要工具。

The retention samples used in comparative studies should include samples that represent marketed product and, when possible, pivotal clinical trial material.

用于对比研究的留样应包括代表上市药品的样品，如可能，还应包括关键的临床试验物料。

If a risk-based evaluation or other drivers lead to changes in an analytical procedure or replacement with a new method or if the procedure is transferred to a new testing site; revalidation, a new validation exercise, an analytical method comparability study, or a combination of these exercises should be considered. In some cases, changes to the drug substance or drug product manufacturing process may also warrant analytical procedure revalidation. These additional studies are discussed below.

如果基于风险的评估或其它原因导致对分析方法进行变更，或采取新的方法取代旧的方法，或分析方法转移至一个新的检测场所，则要考虑进行再验证、新的验证、分析方法对比研究或联合进行这些工作。在有些情形下，对原料药或药品生产工艺的变更也会导致分析方法再验证。这些额外的研究讨论如下：

A. Revalidation 再验证

Principles described in the validation section (section VI) apply to revalidation. When a change is made to an analytical procedure (e.g., a change in a piece of equipment or reagent or because of a change in manufacturing process or formulation), revalidation of all or part of the analytical procedure should be considered. Analytical method revalidation may also be warranted because of manufacturing process changes, such as an alteration in the drug substance manufacturing process that could impact method performance (e.g., route of synthesis, fermentation) or introduction of a new drug product formulation.

在验证部分（第VI部分）所述原则适用于再验证。如果对一个分析方法进行了变更（例如，对设备有变更，或试剂的变更，或因为生产工艺或配方有变更），则可能要考虑对分析方法进行全部或部分再验证。在和平工艺变更时可能也需要对分析方法进行再验证，例如可能影响分析方法性能的原料药生产工艺变更（例如，合成路线、发酵）或引入新的制剂配方。

You should revalidate to ensure that the analytical procedure maintains its critical performance characteristics (e.g., specificity, precision, accuracy). The degree of revalidation depends on the nature of the change.

你要进行再验证以保证分析方法维持其关键性能指标（例如，专属性、精密度、准确性）。再验证的程度取决于变更的性质。

B. Analytical Method Comparability Studies 分析方法对比研究

Analytical method comparability study requests are typically generated when you propose to substitute an FDA-approved analytical procedure with an alternative analytical procedure or when an analytical method is transferred from one laboratory to the other. For information on statistical procedures to use for determining equivalence of two test methods, appropriate literature or text should be consulted[19].These scenarios are discussed below.

分析方法对比研究要求一般是在你提议采用一个替代分析方法取代一个FDA批准的分析方法时，或将一个分析方法从一个实验室转移至另一个实验室时产生的。用于决定两个分析方法的等同性的统计学方法信息，需要引用适当的文献或文件。这些情况讨论如下：

1. Alternative Analytical Procedures 可替代的分析方法

An alternative analytical procedure is an analytical procedure that you use in place of the FDAapproved analytical procedure. For an NDA or ANDA, you should include any proposed alternate analytical procedures in the application. You must include a description of theprocedure[20].After approval, for an NDA or ANDA, or for a procedure approved in a BLA butnot included in an FDA regulation, the addition, revision, or deletion of an alternative analyticalprocedure that provides the same or increased assurance of the identity, strength, quality, purity, or potency of the material being tested as the analytical procedure described in the approved application, must be documented in the next annual report[21].

替代性分析方法是你用来代替FDA已经批准的分析方法的一种分析方法。对于一个NDA或ANDA，你要将所有拟定的替代分析方法包括在申报资料中。你必须包括方法描述。在批准后，对于一个NDA或ANDA，或在BLA里批准但未包括在FDA法规里的分析方法，凡增加、修改或删除替代分析方法均要在下一次年报中记载。

For biological products, in rare cases an analytical procedure may be included in an FDA regulation. If the analytical method required is described by a regulation, however, and you wantto use an alternate method, you must submit the alternate method for review and approval according to 21 CFR 610.9(a). You must present evidence “…demonstrating that the modification will provide assurances of the safety, purity, potency, and effectiveness of the biological product equal to or greater than the assurances provided by the method or process specified in the general standards or additional standards for the biological product.” Modification of such procedures requires FDA approval during application review or in a postapproval supplement[22].

对于生物制品，FDA法规里可能很少包括分析方法。如果所需的分析方法在法规里进行了描述，但你想使用一个方法来替代，你必须根据21 CFR 610.9(a)提交替代方法供审核和批准。你必须提交证据“……证明方法修订能确保生物制品的安全、纯度、效价和有效性等同或优于生物制品通用标准或附加标准中给出的方法或程序”。对这样程序的修订需要在申报资料评估过程中或在批准后增补中获得FDA批准。

You should identify the use of the alternative analytical procedure (e.g., release, stability testing) and provide a rationale for its inclusion, validation data, and comparative data to the FDA-approved analytical procedure. You should perform an analytical method comparability study that demonstrates at a minimum that:

你要分辨可替代性分析方法的使用（例如，放行检测、稳定性测试），提供其内容的合理性论证、验证数据和与FDA批准的分析方法的对比数据。你要进行分析方法对比研究，至少证明：

?   The new method coupled with any additional control measures is equivalent orsuperior to the original method for the intended purpose.

?   新的方法配备了另外的控制手段，在其既定用途上等同或超过原始方法

?   The new analytical procedure is not more susceptible to matrix effects than theoriginal procedure.

?   新的分析方法比原始方法更不易受到基质的影响

If new process-related or product-related variants or any new impurities are discovered with the new procedure, testing on retention samples from historical batches should be performed to demonstrate that the variants/impurities detected by the new method are a result of an increase in the sensitivity or selectivity of the new procedure and not a result of a change to process-related impurities.

如果采用新的检验方法能发同新的与工艺杂质或产品变化产生的杂质或所有新的杂质，则应对历史批准的留样进行检查，证实新方法检出的变化/杂质是因为新的方法灵敏度或选择性增加的结果，而不是工艺杂质变化的结果。

If the procedure has stability-indicating properties:

如果分析方法具有稳定性指示特性：

?   Appropriate samples should be included that allow a comparison of the ability ofthe new and original method to detect relevant product variants and degradation species.

?   应包括适当的样品，比较新的方法和原始方法检出相关产品变化和降解物的能力

?   The number of batches analyzed for comparison should provide sufficient statistical power.

?   对比时所分析的批次数应能提供足够的统计功效

?   Equivalence, non-inferiority, or superiority studies should be performed with appropriate statistical methods to demonstrate that the new or revised methods performance is comparable or better than the original method[23].

?   要采用适当的统计学方法来实施等同性、不低于或优越性研究，来证明新方法或修订过的方法的性能等同或优于原始方法

?   The statistical analyses performed to compare product testing should be identified.

?   要识别出用于比较产品检测结果的统计学分析方法

?   All bias or differences between analytical procedures seen with comparative results should be discussed with an explanation, as appropriate.

?   适当时，分析方法间所有观察到的偏差或差异以及对比结果均应进行讨论，并提出解释

2. Analytical Methods Transfer Studies 分析方法转移研究

Analytical method transfer is typically managed under a transfer protocol that details the parameters to be evaluated in addition to the predetermined acceptance criteria that will be applied to the results. Transfer studies usually involve two or more laboratories or sites (originating lab and receiving labs) executing the preapproved transfer protocol. A sufficient number of representative test articles (e.g., same lot(s) of drug substance or drug product) are used by the originating and receiving laboratories. The comparative studies are performed to evaluate accuracy and precision, especially with regard to assessment of inter laboratory variability. In cases where the transferred analytical procedure is also a stability-indicating method, forced degradation samples or samples containing pertinent product-related impurities should be analyzed at both sites. The USP General Chapter <1224>Transfer of Analytical Procedures provides additional guidance on this topic.

分析方法转移一般要使用转移方案进行管理，在方案中详细写明要评估的参数，以及适用于结果的预定可接受标准。转移研究通常包括两个或更多化验室或场所（转出化验室和接收化验室），由其实施预先批准的转移方案。转出化验室和接收化验室使用具有代表性的足够数量的测试物（例如，相同批号的原料药或制剂）。实施对比研究是为了评估准确度和精密度，特别是实验室之间的差异。如果所转移的分析方法也是稳定性指示性方法，则应在两个化验室均对强降解样品或含有所要检测的相关杂质的样品进行检测。USP通论<1224>“分析方法转移”提供了关于此问题的更多指南。

C. Reporting Postmarketing Changes to an Approved NDA, ANDA, or BLA 已批准的NDA、ANDA或BLA的上市后变更报告

Postmarketing changes to analytical procedures must be reported to the FDA in compliance with 21 CFR 314.70 or 21 CFR 601.12[24].Additional information on the appropriate reporting category for various kinds of postapproval changes for NDAs and ANDAs is provided in the FDA guidance for industry on Changes to an Approved NDA or ANDA and Changes to an Approved NDA or ANDA; Specifications – Use of Enforcement Discretion for CompendialChanges. Similar information on postapproval changes to BLAs regulated by CDER and CBERis provided in the FDA guidance Changes to an Approved Application for Specified Biotechnology and Specified Synthetic Biological Products.

上市后对分析方法的变更必须根据 21 CFR 314.70 或 21 CFR 601.12向FDA报告。在FDA行业指南“已批准NDA或ANDA变更”和“已批准NDA或ANDA变更：质量标准---药典变更自行裁定实施”中给出了NDA和ANDA各种批准后变更的报告分类信息。由CDER和CBER管理的BLA上市后变更的类似信息已在FDA指南“特定生物技术和特定合成生物药品已批准申报的变更”中给出。

IX. FDA METHODS VERIFICATION /FDA方法确认

Part of the approval process for NDAs and ANDAs may include FDA laboratory assessment to determine whether the analytical procedures are acceptable for quality control and suitable for regulatory purposes[25].If a laboratory assessment will be conducted, the FDA laboratory will send you a request that will detail what samples and supplies to send to the FDA laboratory. These could include product samples, standards, critical reagents, material safety data sheets, and supplies. Laboratory results and comments will be forwarded from the FDA laboratory to the product quality reviewer.

有些NDA和ANDA的批准过程会包括FDA实验室评估决定一个分析方法是否被接受作为质量控制用，是否适合于法规目的。如果实施了化验室评估，则FDA实验室会向你发出一个要求，在其中详细说明要呈送给FDA实验室的样品和备用样品。其中可能包括产品样品、对照品、关键的试剂、物料安全数据、以及备用样品。实验室结果和建议会从FDA实验室送至产品质量审核人员那里。

For certain biological products, samples representative of the product for licensure along with summaries of results of tests performed on the lots represented by these samples should be submitted with the BLA[26].The FDA laboratory verifies the performance of the methods and the results you submit. During a pre-BLA meeting or after submission of the BLA, the FDA laboratory can send you a request to provide standards, controls, reagents, material safety data sheets, and supplies.

对于特定的生物药品，则要和BLA一起提交具有代表性的产品样品，以及对这些批次样品检验的结果汇总。FDA实验室会核实这些方法的性能以及你提交的结果。在BLA准备提交时的会议中，或在BLA提交之后，FDA实验室可能会发给你一个要求，要求提供对照品、受控样品、试剂、MSDS和备用样品。

X. REFERENCES 参考文献

Guidance for Industry[27]行业指南

?  ANDAs: Impurities in Drug Products (November 2010)

?  ANDA：制剂中的杂质

?  ANDAs: Impurities in Drug Substances (July 2009)

?  ANDA：原料药中的杂质

?  Changes to an Approved NDA or ANDA (April 2004)

?  已批准NDA或ANDA的变更

?  Changes to an Approved Application for Specified Biotechnology and Specified Synthetic Biological Products (July 1997)

?  已批准特定生物技术和特定合成生物药品申报的变更

?  Changes to an Approved NDA or ANDA; Specifications – Use of Enforcement Discretion for Compendial Changes (November 2004)

?  已批准NDA或ANDA的变更---质量标准---对药典变更行使自由裁量

?  Content and Format of Investigational New Drug Applications (INDs) for Phase 1 Studies of Drugs, Including Well-Characterized, Therapeutic, Biotechnology-derived Products (November 1995)

?  药品一期临床研究IND内容和格式

?  IND Meetings for Human Drugs and Biologics, Chemistry Manufacturing and Controls Information (May 2001)

?  人药和生物制品，研发生产控制IND会议

?  INDs for Phase 2 and 3 Studies of Drugs, Including Specified Therapeutic Biotechnology-Derived Products (February 1999)

?  药品二期和三期临床研究IND，包括特定疗法生物制品衍生产品

?  Investigating Out of Specification (OOS) Test Results for Pharmaceutical Production (October 2006)

?  药品生产结果OOS调查，

?  Process Validation: General Principles and Practices (January 2011)

?  工艺验证一般原则和规范

?  Reviewer Guidance, Validation of Chromatographic Methods (November 1994)

?  审核指南，色谱方法验证

?  Submission of Chemistry, Manufacturing, and Controls Information for Synthetic Peptide Substances (November 1994)

?  合成多肽物CMC信息申报

Guidance for Industry: International Conference on Harmonization

行业指南：ICH

?  Q1A(R2) Stability Testing of New Drug Substances and Products (November 2003)

?  新原料药和制剂的稳定性测试

?  Q1B Stability Testing: Photostability Testing of New Drug Substances and Products (May 548 1997)

?  稳定性测试：新原料药和制剂的光照试验

?  Q1C Stability Testing for New Dosage Forms (May 1997)

?  新剂型的稳定性测试

?  Q2(R1) Validation of Analytical Procedures: Text and Methodology (March 1995, May 1997)

?  分析方法验证：正文和方法学

?  Q3A(R2) Impurities in New Drug Substances (June 2008)

?  新原料药中的杂质

?  Q3B(R2) Impurities in New Drug Products (August 2006)

?  新制剂中的杂质

?  Q3C Impurities: Residual Solvents (December 1997)

?  杂质：残留溶剂

?  Q3C Tables and List (February 2012)

?  表格和清单

?  Q5C Quality of Biotechnological Products: Stability Testing of Biotechnological/Biological Products (July 1996)

?  生物技术药品的质量：生物技术/生物制品稳定性测试

?  Q6A Specifications: Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances (December 2000)

?  质量标准：新原料药和新制剂的检验方法和可接受标准：化学物质

?  Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products (August 1999)

?  质量标准：生物技术/生物制品检验方法和可接受标准

?  Q7 Good Manufacturing Practice Guide for Active Pharmaceutical Ingredients (August 2001)

?  原料药GMP

United States Pharmacopeia/National Formulary 美国药典处方集

?  General Chapter <621> Chromatography

?  通论<621>色谱

?  General Chapter <1010> Analytical Data – Interpretation and Treatment

?  通论<1010>分析数据---诠释和处理

?  General Chapter <1224> Transfer of Analytical Procedures

?  通论<1224>分析方法转移

?  General Chapter <1225> Validation of Compendial Procedures

?  通论<1225>药典方法验证

?  General Chapter <1226> Verification of Compendial Procedures

?  通论<1226>药典方法确认

?  General Notices and Requirements, Applying to Standards, Tests, Assays, and Other Specifications of the United States Pharmacopeia: 7. Test Results

?  通则，适用于标准、测试、含量和其它USP质量标准，7，检测结果

?  Interpretation and Treatment of Analytical Data; USP Pharmacopeial Forum, United States Pharmacopeial Convention, Inc., Rockville MD: 1994, Volume 24, Number 5, pp. 7051 - 7056

?  分析数据的诠释和处理，USP药典论坛，美国药典委员会公司

Other 其它

?  ASTM Standard, E29 - 2008 Standard Practice for Using Significant Digits in Test Data to Determine Conformance with Specifications, ASTM International, West Conshohocken, PA, (www.astm.org).

?  ASTM标准，E29-2008，检验数据中使用有效数位来决定与标准的符合性的标准规范

?  ASTM E1488 – Standard Guide for Statistical Procedures to use in Developing and Applying Test Methods, ASTM International, West Conshohocken, PA, (www.astm.org).  

?  ASTM E1488—用于研发和使用检验方法的统计学程序标准指南

?  ASTM E2782 – Standard Guide for Measurement Systems Analysis (MSA), ASTMInternational, West Conshohocken, PA, (www.astm.org).  

?  ASTM E2782—测量系统分析标准指南

?  ASTM Standard, E2935 – 2013 Standard Practice for Conducting Equivalence Testing in Laboratory Applications, ASTM International, West Conshohocken, PA, (www.astm.org).

?  ASMT标准E2935-2013，实验室应用中进行等同性测试标准规范

?  J.N. Miller and Miller, J.C., 2010, Statistics and Chemometrics for Analytical Chemistry, 6th edition, Pearson Education Canada.

?  J.N.MILLER和MILLER,J.C，2010，分析化学统计学和化学计量学，第6版

?  Saunders, B.D. and R.G. Trapp, 2004, Basic and Clinical Biostatistics, 4th edition, Lange Medical Books/McGraw Hill.

?  SAUNDERS，B.D.和R.G.TRAPP，2004，基础和临床生物统计，第4版
, d! v9 S8 e8 l2 d% x' B' k2 r

学习学习，谢谢分享