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

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16 9467) and replaces the 2000 draft guidance for industry on Analytical Procedures and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 onINDs 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。