Analytical Procedures and Methods Validation for Drugs and Biologics
药品和生物制品分析方法验证
Guidance for Industry
行业指南[1]
U.S. Department of Health and Human Services
Food and Drug Administration
Center for Drug Evaluation and Research (CDER)
Center for Biologics Evaluation and Research (CBER)
July 2015
Pharmaceutical Quality/CMC
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guidance supersedes the draft of the same name that published on February 19, 2014 (79 FR169467) and replaces the 2000 draft guidance for industry onAnalytical Procedures and Methods Validation[2][3][4][5][6]
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) guidanceQ2(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]
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 onProcess 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 wordshouldin 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][9][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 ICHM2 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 theUnited 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]
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 onValidation of Chromatographic Methodsand 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。
