美国FDA分析方法验证指南中文译稿[1]

I. 导言……………………………………………………………………………… 1 II. 背景……………………………………………………………………………… 2 III. 分析方法的类型………………………………………………………………… 3 A. 法定分析方法……………………………………………………………. 3 B. 可选择分析方法…………………………………………………………. 3 3 C. 稳定性指示分析…………………………………………………………. 3 IV. 对照品…………………………………………………………………………… 4 A. 对照品的类型……………………………………………………………. 4 B. 分析报告单………………………………………………………………. 4 C. 对照品的界定……………………………………………………………. 4

V. IND 中的分析方法验证………………………………………………………… 6 VI. NDA, ANDA, BLA 和PLA 中分析方法验证的内容和格式…………………. 6 A. 原则………………………………………………………………………. 6

B. 取样………………………………………………………………………. 7 C. 仪器和仪器参数…………………………………………………………. 7 D. 试剂………………………………………………………………………. 7 E. 系统适应性实验…………………………………………………………. 7 F. 对照品的制备……………………………………………………………. 7 G. 样品的制备………………………………………………………………. 8 H. 分析方法…………………………………………………………………. 8 L. 计算………………………………………………………………………. 8 J. 结果报告…………………………………………………………………. 8

VII. NDA，ANDA,BLA 和PLA 中的分析方法验证………………………………. 9

A． 非法定分析方法………………………………………………………… 9 1． 验证项目……………………………………………………………. 9 2. 其它分析方法验证信息……………………………………………. 10

a. 耐用性………………………………………………………… 11 b. 强降解实验…………………………………………………… 11 c. 仪器输出/原始资料………………………………………… 11 3． 各类检测的建议验证项目…………………………………………. 13 B． 法定分析方法…………………………………………………………… 15 VIII. 统计分析………………………………………………………………………… 15

A. 总则………………………………………………………………………. 15 B. 比较研究…………………………………………………………………. 16 C. 统计………………………………………………………………………. 16

IX. 再验证…………………………………………………………………………… 16 X. 分析方法验证技术包：内容和过程…………………………………………… 17 A. 分析方法验证技术包…………………………………………………… 17 B. 样品的选择和运输……………………………………………………… 18

C. 各方责任…………………………………………………………………. 19

XI. 方法……………………………………………………………………………… 20 A. 高效液相色谱(HPLC) …………………………………………………… 20 B. 气相色谱(GC) …………………………………………………………… 22 C. 分光光度法，光谱学，光谱法和相关的物理方法……………………. 23 D. 毛细管电泳………………………………………………………………. 23 E. 旋光度……………………………………………………………………. 24 F. 粒径相关的分析方法…………………………………………………… 25 G. 溶出度…………………………………………………………………… 26 H. 其它仪器分析方法……………………………………………………… 27 附

件

A

：

NDA

，

ANDA

，

BLA

和

PLA

申

请

的

内

容………………………………………… 28

附件 B：分析方法验证的问题和延误…………………………………………………… 29

参考文献……………………………………………………………………………………

30

术语表……………………………………………………………………………………… 32

This guidance provides recommendations to applicants on submitting analytical

procedures, validation data, and samples to support the documentation of the identity,

strength, quality, purity, and potency of drug substances and drug products. 1. 绪论

本指南旨在为申请者提供建议，以帮助其提交分析方法，方法验证资料和样品用于支持

原料药和制剂的认定，剂量，质量，纯度和效力方面的文件。

This guidance is intended to assist applicants in assembling information, submitting

samples,

and

presenting

data

to

support

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), product license applications (PLAs), and supplements to these

applications.

本指南旨在帮助申请者收集资料，递交样品并资料以支持分析方法。这些建议适用于NDA，

ANDA，BLA，PLA及其它们的补充中所涉及的原料药和制剂。

The principles also apply to drug substances and drug products covered in Type

II drug master files (DMFs). If a different approach is chosen, the applicant is encouraged to discuss the matter in advance with the center with product jurisdiction to prevent the expenditure of resources on preparing a submission that

may later be determined to be unacceptable.

这些原则同样适用于二类DMF所涉及的原料药和制剂。如果使用了其它方法，鼓励申请者

事先和FDA药品评审中心的进行讨论，以免出现这种情况，那就是花了人力物力所准备起

来的递交资料后来发现是不可用的。

The principles of methods validation described in this guidance apply to all types

of analytical procedures. However, the specific recommendations in this guidance may

not be applicable to certain unique analytical procedures for products such as

biological, biotechnological, botanical, or radiopharmaceutical drugs. 本指南中所述的分析方法验证的原则适用于各种类型的分析方法。但是，本指南中特定

的建议可能不适用于有些产品所用的特殊分析方法，如生物药，生物技术药，植物药或放射

性药物等。

For example, many bioassays are based on animal challenge models, 39 immunogenicity assessments, or other immunoassays that have unique

features that

should be considered when submitting analytical procedure and methods validation

information.

比如说，许多生物分析是建立在动物挑战模式，免疫原性评估或其它有着独特特性的免

疫分析基础上的，在递交分析方法和分析方法验证资料时需考虑这些独特的性质。 Furthermore, specific recommendations for biological and immunochemical tests that

may be necessary for characterization and quality control of many drug substances

and drug products are beyond the scope of this guidance document. 而且，许多原料药和制剂的界定和质量控制所需的生物和免疫化学检测并不在本指南的

范围之内。

Although this guidance does not specifically address the submission of analytical

procedures and validation data for raw materials, intermediates, excipients, container closure components, and other materials used in the production of drug

substances and drug products, validated analytical procedures should be used to

analyze these materials.

尽管本指南并不专门叙述原料，中间体，赋形剂，包装材料及原料药和制剂生产中所用

的其它物料的分析方法及分析方法验证资料的递交，但是应该应用验证过的分析方法来分析

检测这些物质。

For questions on appropriate validation approaches for analytical procedures or

submission of information not addressed in this guidance, applicants should consult

with the appropriate chemistry review staff at FDA.

对于本指南中未提及的关于分析方法验证和资料提交方面的问题，请向FDA相关的

化学评

审人员咨询。

This guidance, when finalized, will replace the FDA guidance for industry on Submitting Samples and Analytical Data for Methods Validation (February 1987).

本指南，一旦定稿，将取代FDA于1987年2月份发布的工业指南：分析方法验证所需提交

的样品和分析资料。

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, including bioavailability of the drug product (21 CFR 314.50(d)(1) and

314.94(a)(9)(i)). II. 背景

每个NDA和ANDA都必需包括必要的分析方法以确保原料药和制剂的认定，剂

量，质量，纯

度和效力，还包括制剂的生物利用度(21 CFR 314.50(d)(1) 和314.94(a)(9)(i))。 Data must be available to establish that the analytical procedures used in testing

meet proper standards of accuracy and reliability (21 CFR 211.165(e) and 211.194(a)(2)).

必须要有资料来论证所用的分析方法是符合一定的准确度和可靠性标准的。 Methods validation is the process of demonstrating that analytical procedures are

suitable for their intended use. The methods validation process for analytical procedures begins with the planned and systematic collection by the applicant of the

validation data to support the analytical procedures.

分析方法验证是论证某一分析方法适用于其用途的过程。分析方法的验证过程是从申请

者有计划地系统性收集验证资料以支持分析方法开始的。

The review chemist evaluates the analytical procedures and validation data submitted in the NDA or ANDA.

审评化学家会对NDA或ANDA中的分析方法和验证资料进行评审。

On request from FDA, an NDA or ANDA applicant must submit samples of drug product,

drug substance, noncompendial reference standards, and blanks so that the applicant's

drug substance and drug product analytical procedures can be evaluated by FDA

laboratories (21 CFR 314.50(e) and 314.94(a)(10)).

一旦FDA有要求，则NDA或ANDA的申请者必须提交制剂，原料药，非药典对照品和空白以

使FDA实验室能对申请者所用分析方法进行评审(21 CFR 314.50(e) and 314.94(a)(10))。

The FDA laboratory analysis demonstrates that the analytical procedures are reproducible by laboratory testing. The review chemists and laboratory analysts

determine the suitability of the analytical procedures for regulatory purposes. FDA实验室的分析会论证该分析方法在实验室内是可以重现的。审评化学家和实验室分析

家会从法规的角度确定该分析方法的适用性。

FDA investigators inspect the analytical laboratory testing sites to ensure that the

analytical procedures used for release and stability testing comply with current good

manufacturing practices (CGMPs) (21 CFR part 211) or good laboratory practices (GLPs)

(21 CFR part 58), as appropriate.

FDA检查官会对分析实验室进行检查确保用于放行和稳定性实验的分析方法符合现行的

GMP（21CFR part 211) 和GLP (21 CFR part 58)。

Each BLA and PLA must include a full description of the manufacturing methods,

including analytical procedures, that demonstrate that the manufactured

product

meets prescribed standards of safety, purity, and potency (21 CFR 601.2(a) and

601.2(c)(1)(iv)).

每个BLA和PLA都必须要有详细的生产工艺描述，包括分析方法，以说明所生产的产品是

符合规定睥安全，纯充和效力标准的(21 CFR 601.2(a) and 601.2(c)(1)(iv))。 Data must be available to establish that the analytical procedures used in testing

meet proper standards of accuracy and reliability (21 CFR 81

211.194(a)(2)). For BLAs, PLAs, and their supplements, the analytical procedures and

their validation are submitted as part of the license application or supplement and

are evaluated by the review committee.

必须要有资料证明所用的分析方法是符合一定的准确度和可靠性要求的(21 CFR 81

211.194(a)(2))。对于BLA，PLA及它们的补充，在所提交的许可证申请中应当要有分析方法

和方法验证这部分的资料，审评委员会会对这部分资料进行评审。

Representative samples of the product must be submitted and summaries of results of

tests performed on the lots represented by the submitted sample must be provided (21

CFR 601.2(a) and 601.2(c)(1)(vi)). The review committee chair may request analytical

testing by CBER laboratory analysts to evaluate the applicant=s analytical procedures

and verify the test results.

需提供代表性样品及该样品所代表批号的检测结果总结(21 CFR 601.2(a) and 601.2(c)(1)(vi))。评审委员会会要求CBER实验室的分析人员进行分析实验对申请者的

分析方法进行评估，并确认其分析结果。

All analytical procedures are of equal importance from a validation

perspective. In

general, validated analytical procedures should be used, irrespective of whether they

are for in-process, release, acceptance, or stability testing. Each quantitative analytical procedure should be designed to minimize assay variation. 从验证的角度来看，所有的分析方法有着同样的重要性。一般来说，应当要应用已验证

过的分析方法，而不论其是被用于过程控制，放行，合格或稳定性实验。高等每个定量分析

方法时都应当要减少其分析误差。

Analytical procedures and validation data are submitted in the sections of the application on analytical procedures and controls. Recommendations on information

to be submitted are included in sections III through IX and XI of this guidance. Information on submission of the methods validation package to the NDA or ANDA and

samples to the FDA laboratories is provided in section X.

分析方法和验证资料应当摆在申请的分析方法和控制章节中提交。本指南的第III到IX

章和XI章给出了所需提供资料方面的建议。向FDA实验室提供样品和递交NDA和ANDA中的分析

方法验证资料的信息见第X章。

A. Regulatory Analytical Procedure A regulatory analytical procedure is the analytical procedure used to evaluate a defined characteristic of the drug substance

or drug product. The analytical procedures in the U.S. Pharmacopeia/National Formulary (USP/NF) are those legally recognized under section 501(b) of the Food,

Drug, and Cosmetic Act (the Act) as the regulatory analytical procedures for compendial items. For purposes of determining compliance with the Act, the regulatory

analytical procedure is used.

III 分析方法的类型 A. 法定分析方法

法定分析方法是被用来评估原料药或制剂的特定性质的。USP/NF中的分析方法是法定的

用于药典项目检测的分析方法。为了确认符合法规，需使用法定分析方法。 B. Alternative Analytical Procedure

An alternative analytical procedure is an analytical procedure proposed by the

applicant for use instead of the regulatory analytical procedure. A validated alternative analytical procedure should be submitted only if it is shown to perform

equal to or better than the regulatory analytical procedure. B. 替代分析方法

替代分析方法是申请者提出用于代替法定分析方法的分析方法。只有当一替代分析方法

相当于或优于法定分析方法时，才可以应用验证过的替代分析方法。

If an alternative analytical procedure is submitted, the applicant should provide

a rationale for its inclusion and identify its use (e.g., release, stability testing), validation data, and comparative data to the regulatory analytical procedure. 如果提交了替代分析方法，申请者还应当提供其理由，并标明其用途（如，放行，稳定

性实验），验证资料及其与法定分析方法的对比资料。 C. Stability-Indicating Assay

A stability-indicating assay is a validated quantitative analytical procedure that

can detect the changes with time in the pertinent properties of the drug substance

and drug product. C. 稳定性指示分析

稳定性指示分析是能检测出原料药或制剂的某些性质随着时间的延长而出现的变化的定

量分析方法。

A stability-indicating assay accurately measures the active ingredients, without

interference from degradation products, process impurities, excipients, or other

potential impurities。

稳定性指示分析能不受降解产物，工艺杂质，赋形剂或其它潜在杂质的影响而准确测定

其中的活性成分。

If an applicant submits a non-stability-indicating analytical procedure for release testing, then an analytical procedure capable of qualitatively and quantitatively monitoring the impurities, including degradation products, should

complement it. Assay analytical procedures for stability studies should be stability-indicating, unless scientifically justified.

如果申请者递交了用于放行检测的非稳定性指示分析方法，则应当要有能定性和定量

地

监测杂质，包括降解产物，的分析方法对其进行补充。稳定性试验中所用的含量分析方法应

当要有稳定性指示能力，除非有科学的理由能证明其合理性。

A reference standard (i.e., primary standard) may be obtained from the USP/NF

or other official sources (e.g., CBER, 21 CFR 610.20). If there are questions on whether a source of a standard would be considered by FDA to be an official source,

applicants should contact the appropriate chemistry review staff. When there is no

official source, a reference standard should be of the highest possible purity and

be fully characterized. IV 标准品 A． 标准品的类型

可以从USP/NF处或其它官方(比如说，CBER，21CFR 610.20)获得标准品 (也就是一级对

照品)。如果不能确定一标准品的来源是否会被FDA认为是官方来源，申请者应当要向适当的

化学评审人员咨询。如果没有官方来源，则被用来作标准品的物质应当要有尽可能高的纯度，

并得到充分界定。

A working standard (i.e., in-house or secondary standard) is a standard that is qualified against and used instead of the reference standard.

工作对照品 (也就是内部标准品或二级标准品)是根据一级对照品标定的，并用来代替一

级对照品的。

A certificate of analysis (COA) for reference standards from non-official sources

should be submitted in the section of the application on analytical procedures and

controls. For standards from official sources, the user should ensure the

suitability

of the reference standard. The standard should be stored correctly and used within

the established use interval. B． 分析报告单

对于非官方标准品，在申请的分析方法和控制章节中应当要提供该标准品的分析报告单。

对于从官方获得的标准品，用户应当要确保标准品的适用性。应当正确储存标准品并在已确

定的时间段内使用该标准品。

Reference standards from USP/NF and other official sources do not require further

characterization. A reference standard that is not obtained from an official source

should be of the highest purity that can be obtained by reasonable effort, and it

should be thoroughly characterized to ensure its identity, strength, quality,

purity,

and potency. C． 标准品的界定

从USP/NF及其它官方来源获得的标准品是不需要进一步界定的。非官方对照品要有尽可

能高的纯度，并进行充分地界定以确保其结构，剂量，质量，纯度和效力。 The qualitative and quantitative analytical procedures used to characterize a reference standard are expected to be different from, and more extensive than, those

used to control the identity, strength, quality, purity, and potency of the drug substance or the drug product. Analytical procedures used to Draft — Not for Implementation characterize a reference standard should not rely solely on comparison

testing to a previously designated reference standard.

用于界定标准品的定性和定量分析方法应当要不同于用于控制原料药或制剂的结构，剂

量，质量，纯度和效力的分析方法，要比它们更深入。用于标准品界定的分析方法不应仅仅

是和先前的指定标准品进行比较实验。

Generally, this characterization information should include:

A brief description of the manufacture of the reference standard, if the manufacturing process differs from that of the drug substance. Any additional

purification procedures used in the preparation of the reference standard should be

described.

一般来说，界定资料应当要包括：

标准品的简单工艺描述，如果其生产工艺是否于其相应的原料药的话。应当要叙述制备

标准品时所用的补充精制过程。

Legible reproductions of the relevant spectra, chromatograms, thin-layer chromatogram (TLC) photographs or reproductions, and other appropriate

instrumental

recordings. Data establishing purity. The data should be obtained by using appropriate tests, such as TLC, gas chromatography (GC), high-pressure liquid

chromatography (HPLC), phase solubility analysis, appropriate thermometric analytical procedures, and others as necessary.

相关光谱图，色谱图，TLC照片及其它仪器输出的清晰复印件。

建立纯度的资料。应当要应用适当的检测方法来获得这些资料，比如说TLC，GC，HPLC，

相溶解分析，适当的热分析方法及其它必要的分析方法。

Appropriate chemical attribute information, such as structural formula, empirical formula, and molecular weight. Information to substantiate the proof of

structure should include appropriate analytical tests, such as elemental analysis,

infrared spectrophotometry (IR), ultraviolet spectrophotometry (UV), nuclear

magnetic resonance spectroscopy (NMR), and mass spectrometry (MS), as well as

applicable functional group analysis. Detailed interpretation of the test data in

support of the claimed structure should be provided.

适当的化学性质资料，比如结构式，经验式和分子量等。结构确证资料应当要包括适当

的分析测试，比如元素分析，IR，UV,NMR和MS，及适用的官能团分析。还应当要提供具体的

结构解析资料。

A physical description of the material, including its color and physical form. Appropriate physical constants such as melting range, boiling range, refractive index,

dissociation constants (pK values), and optical rotation. A detailed description