ICH指导原则之方法论.doc

ICH Q2 (R1)
INTRODUCTION 简介
This document is complementary to the parent document, which presents a discussion of the characteristics that should be considered during the validation of analytical procedures. Its purpose is to provide some guidance and recommendations on how to consider the various validation characteristics for each analytical procedure. In some cases (for example, demonstration of specificity), the overall capabilities of a number of analytical procedures in combination may be investigated in order to ensure the quality of the drug substance or drug product. In addition, the document provides an indication of the data, which should be presented in a registration application.
本文作为前文的补充，旨在讨论在分析方法验证过程中在每一个具体的项目需要考虑哪些内容。本文的目的是就不同类型的验证该涵盖哪些项目提供一个指导原则和建议。以专属性为例，为了确保原料药或制剂的质量，需要考察分析过程对化合物中杂质的全面综合分析能力。另外，文件提供的数据应该包含在注册申请材料中。

All relevant data collected during validation and formulae used for calculating validation characteristics should be submitted and discussed as appropriate.
验证报告中的所有数据及每个验证项目的计算公式应一同提交并进行适当的论述(即得出相应结论)。

Approaches other than those set forth in this guideline may be applicable and acceptable. It is the responsibility of the applicant to choose the validation procedure and protocol most suitable for their product. However it is important to remember that the main objective of validation of an analytical procedure is to demonstrate that the procedure is suitable for its intended purpose. Due to their complex nature, analytical procedures for biological and biotechnological products in some cases may be approached differently than in this document.
本文阐述的原则之外的原则应该适用并可接受。申请者的职责是制定最适合申报产品的验证项目及验证方案。最重要的是记住分析方法验证的主要目的是证明该分析程序能达到预期目标。由于生物制品和生物科技产品本身的复杂性，有时候，其分析方法的验证也可能与本文提到的方法不同。

Well-characterized reference materials, with documented purity, should be used throughout the validation study. The degree of purity necessary depends on the intended use.
验证研究用到的参比物质需要经过完全鉴定并标定纯度。所需的纯度取决于其应用类型(预期用途)。

In accordance with the parent document, and for the sake of clarity, this document considers the various validation characteristics in distinct sections. The arrangement of these sections reflects the process by which an analytical procedure may be developed and evaluated.
与前文相同，该文件考虑了各个独立章节的不同验证项目。这些章节的安排也反映了分析方法的建立和评估的过程。

In practice, it is usually possible to design the experimental work such that the appropriate validation characteristics can be considered simultaneously to provide a sound, overall knowledge of the capabilities of the analytical procedure, for instance: specificity, linearity, range, accuracy and precision.
事实上，设计试验的时候，一些适当的验证项目可以同时考虑，以便对分析方法的能力提供合理的，全面的了解(依据)，例如：专属性、线性、范围、准确度、精密度。
provide a……knowledge of 为……提供依据

1. SPECIFICITY 专属性
An investigation of specificity should be conducted during the validation of identification tests, the determination of impurities and the assay. The procedures used to demonstrate specificity will depend on the intended objective of the analytical procedure.
鉴别测试、杂质和含量测试方法的验证都应考察方法的专属性。证明专属性的方法取决于分析方法的预期目的。

It is not always possible to demonstrate that an analytical procedure is specific for a particular analyte (complete discrimination). In this case a combination of two or more analytical procedures is recommended to achieve the necessary level of discrimination.
一般来说，某一种分析方法不太可能完全证明其对某一特定被分析物具有专属性。这种情况下，建议采用两种或两种以上的分析方法以确保完全鉴别水平。

1.1. Identification 鉴别
Suitable identification tests should be able to discriminate between compounds of closely related structures, which are likely to be present. The discrimination of a procedure may be confirmed by obtaining positive results (perhaps by comparison with a known reference material) from samples containing the analyte, coupled with negative results from samples, which do not contain the analyte. In addition, the identification test may be applied to materials structurally similar to or closely related to the analyte to confirm that a positive response is not obtained. The choice of such potentially interfering materials should be based on sound scientific judgment with a consideration of the interferences that could occur.
合适的鉴别方法应该能够区分可能存在的结构相近的化合物。可以同已知参考物质进行比较，从含有被分析物的样品得到的正的结果和不含被分析物的样品得到的负的结果来确定。此外，鉴别测试也可以用结构相近或相关的物质测试得不到正的反应来证实。在考虑可能会造城干扰的前提下，应根据合理科学的判断来选择可能存在的干扰物。
likely to be present：可能存在的；discrimination：比较；

1.2. Assay and Impurity Test(s) 含量和杂质测定
For chromatographic procedures, representative chromatograms should be used to demonstrate specificity and individual components should be appropriately labelled. Similar considerations should be given to other separation techniques.
在色谱法测定中，需要用有代表性的图谱证明专属性，并恰当的注明每一个成分。其它的分离技术也应如此。

Critical separations in chromatography should be investigated at an appropriate level. For critical separations, specificity can be demonstrated by the resolution of the two components, which elute closest to each other.
色谱分离法应在一定程度上考察关键性的分离。对关键性的分离，可用两个洗脱程度最接近的化合物的分离度来证明其专属性。

In cases where a non-specific assay is used, other supporting analytical procedures should be used to demonstrate overall specificity. For example, where a titration is adopted to assay the drug substance for release, the combination of the assay and a suitable test for impurities can be used.
当采用非专属性的方法测定含量时，应采用辅助性分析方法来证明整个方法具有专属性。例如用滴定法测定放行原料药的含量，可结合使用合适的杂质检测方法。
supporting analytical procedures：辅助性分析方法；overall specificity：整体专属性；combination：结合，联合；

The approach is similar for both assay and impurity tests:
下述方法均适用于含量和杂质检测。
is similar for：适用于

1.2.1 Impurities are available 可以得到杂质的情况
For the assay, this should involve demonstration of the discrimination of the analyte in the presence of impurities and/or excipients; practically, this can be done by spiking pure substances (drug substance or drug product) with appropriate levels of impurities and/or excipients and demonstrating that the assay result is unaffected by the presence of these materials (by comparison with the assay result obtained on unspiked samples).

对含量检测，专属性应该包括提供被分析物在杂质和/或赋形剂存在时能被区分的证明；实际操作时，可通过向纯物质(原料药或制剂)中加入一定量的杂质和/或赋形剂的检测结果和未添加杂质和/或赋形剂的纯物质的检测结果进行对比以此证明这些杂质和/或赋形剂的存在不会对含量检测结果造成影响。
discrimination：区分

For the impurity test, the discrimination may be established by spiking drug substance or drug product with appropriate levels of impurities and demonstrating the separation of these impurities individually and/or from other components in the sample matrix.
对杂质检测，可以向原料药或制剂中加入一定量的杂质，证明各杂质能够分离且能与样品中的其它组分分离。

1.2.2 Impurities are not available 无法得到杂质的情况
If impurity or degradation product standards are unavailable, specificity may be demonstrated by comparing the test results of samples containing impurities or degradation products to a second well-characterized procedure e.g.: pharmacopoeial method or other validated analytical procedure (independent procedure). As appropriate, this should include samples stored under relevant stress conditions: light, heat, humidity, acid/base hydrolysis and oxidation.
如果无法得到杂质或降解产物的对照品，检测方法的专属性可以通过将含有一定量的杂质或降解产物的样品的检测结果与另一种成熟的检测方法——如药典方法或经验证的其它方法(独立的方法)——的检测结果进行比较来证明。必要时，应该包括放置在强降解试验条件，即强光，高温，高湿，酸/碱水解及氧化条件下的样品测试。
As appropriate：必要时；

- For the assay, the two results should be compared;
- 对含量检测，需要对比两种方法的检测结果
- For the impurity tests, the impurity profiles should be compared.
- 对杂质检测，需要对比杂质概况

Peak purity tests may be useful to show that the analyte chromatographic peak is not attributable to more than one component (e.g., diode array, mass spectrometry).
峰纯度测试是非常有用的，它能显示被测物的色谱峰是一个成分还是多个成分(如二极管阵列，质谱)。

2. LINEARITY 线性
A linear relationship should be evaluated across the range (see section 3) of the analytical procedure. It may be demonstrated directly on the drug substance (by dilution of a standard stock solution) and/or separate weighings of synthetic mixtures of the drug product components, using the proposed procedure. The latter aspect can be studied during investigation of the range.
检测方法的线性关系应该在范围内(见章节3)进行评价。线性研究可通过所建议的分析方法，直接对原料药(用标准储备液稀释)和/或分别称取制剂组分的混合物测试来进行。后者应在方法的范围内进行研究。

Linearity should be evaluated by visual inspection of a plot of signals as a function of analyte concentration or content. If there is a linear relationship, test results should be evaluated by appropriate statistical methods, for example, by calculation of a regression line by the method of least squares. In some cases, to obtain linearity between assays and sample concentrations, the test data may need to be subjected to a mathematical transformation prior to the regression analysis. Data from the regression line itself may be helpful to provide mathematical estimates of the degree of linearity.
线性应关系应以信号对被测物浓度或含量作图，根据图形是否呈线性来评估。如果呈线性关系，测试结果应用适当的统计学方法进行评估，例如用最小二乘法进行线性回归计算。在某些情况下，为了使含量与样品浓度呈线性关系，在回归分析前需要对测试数据进行数学转化。由线性回归评估所得的数据本身又有助于精确的评价线性的程度。
In some cases：在某些情况下，有时候；

The correlation coefficient, y-intercept, slope of the regression line and residual sum of squares should be submitted. A plot of the data should be included. In addition, an analysis of the deviation of the actual data points from the regression line may also be helpful for evaluating linearity.
相关系数，y轴上的截距，回归曲线的斜率以及剩余方差应包含在递交材料里。还应包括数据图表。另外，实际数据点与回归曲线的偏差也有助于对线性进行评价。

Some analytical procedures, such as immunoassays, do not demonstrate linearity after any transformation. In this case, the analytical response should be described by an appropriate function of the concentration (amount) of an analyte in a sample.
一些分析方法，如免疫测定法，在任何转换后，均不能证明呈线性。在这种情况下，分析的相应值应用被分析物的浓度(数量)的适当函数来表示。

For the establishment of linearity, a minimum of 5 concentrations is recommended Other approaches should be justified.
为建立线性，建议至少用5个浓度。若用其他方法应证明其合理性。

3. RANGE 范围
The specified range is normally derived from linearity studies and depends on the intended application of the procedure. It is established by confirming that the analytical procedure provides an acceptable degree of linearity, accuracy and precision when applied to samples containing amounts of analyte within or at the extremes of the specified range of the analytical procedure.
特定的范围一般是从线性研究中得到的，它依赖于分析方法的应用目的。确定范围的方法是：样品中含有被分析物的量在分析方法规定的范围内或在范围末端，该分析方法均能获得良好的线性，精密度及准确度。

The following minimum specified ranges should be considered:
以下是应考虑的最小规定范围：
- for the assay of a drug substance or a finished (drug) product: normally from 80 to 120 percent of the test concentration;
- 对原料药或成品药(制剂)的含量测定：一般应在测试浓度的80%～120%；

- for content uniformity, covering a minimum of 70 to 130 percent of the test concentration, unless a wider more appropriate range, based on the nature of the dosage form (e.g., metered dose inhalers), is justified;
- 对含量均匀度检测：应至少在测试浓度的70%～130%之内，超出此范围，应有正当理由，主要是根据剂型的特点(如定量吸入剂)；

- for dissolution testing: +/-20 % over the specified range;
- 对溶出度测试，应为规定范围的+/-20%；

e.g., if the specifications for a controlled released product cover a region from 20%, after 1 hour, up to 90%, after 24 hours, the validated range would be 0-110% of the label claim.
例如：如果是控释剂，规定1小时后达到20%，24小时后达到90%，它的验证范围应为标示量的0～110%。

- for the determination of an impurity: from the reporting level of an impurity1 to 120% of the specification;
- 对杂质测定，应为杂质的报告水平至标准规定的120%；

- for impurities known to be unusually potent or to produce toxic or unexpected pharmacological effects, the detection/quantitation limit should be commensurate with the level at which the impurities must be controlled;
- 对已知有异常功效的，有毒的或者有意外药理作用的杂质，其检测限度和定量限度应与该杂质必须被控制的水平相当。

Note: for validation of impurity test procedures carried out during development, it may be necessary to consider the range around a suggested (probable) limit.
注意：在研制阶段进行杂质检测方法验证时，有必要根据建议(可能)的限度水平来考虑范围；

- if assay and purity are performed together as one test and only a 100% standard is used, linearity should cover the range from the reporting level of the impurities1 to 120% of the assay specification.
1 see chapters “Reporting Impurity Content of Batches” of the corresponding ICH-Guidelines: “Impurities in New Drug Substances” and “Impurities in New Drug Products”
- 如果一个试验同时进行含量和纯度检测，且仅使用100%的标准品，线性范围应覆盖杂质的报告水平(见相应ICH指南“新原料药中的杂质”和“新制剂中的杂质”中“批杂质含量的报告”章节)至含量指标120%。

4. ACCURACY 准确度
Accuracy should be established across the specified range of the analytical procedure.
应在分析方法规定的范围内建立(考察方法的)准确度。
4.1. Assay 含量
4.1.1 Drug Substance 原料药
Several methods of determining accuracy are available:
以下几种方法可用于测定准确度：
a) application of an analytical procedure to an analyte of known purity (e.g. reference material);
用该分析方法测定已知纯度的被分析物(例如参照物质)；

b) comparison of the results of the proposed analytical procedure with those of a second well-characterized procedure, the accuracy of which is stated and/or defined (independent procedure, see 1.2.);
用建议采用的分析方法的结果与另一种完全验证过的方法的结果作对比，对比的方法的准确度是规定的(一定的)和/或已定义的(独立的方法，见1.2节)

c) accuracy may be inferred once precision, linearity and specificity have been established.
准确度可以在精密度，线性及专属性建立之后推论得到；

4.1.2 Drug Product 制剂
Several methods for determining accuracy are available:
以下几种方法可用于测定准确度：
a) application of the analytical procedure to synthetic mixtures of the drug product components to which known quantities of the drug substance to be analysed have been added;
用该分析方法测定按处方量制成的混合物，其中加入了已知量的待测原料药。

b) in cases where it is impossible to obtain samples of all drug product components , it may be acceptable either to add known quantities of the analyte to the drug product or to compare the results obtained from a second, well characterized procedure, the accuracy of which is stated and/or defined (independent procedure, see 1.2.);
如果不能得到制剂的所有成分，向制剂中加入已知量的被测物或者与另一种经过完整验证过的准确度是规定的(一定的)和/或已定义的方法的结果作对比，(独立的方法，见1.2节)也是可以接受的；

c) accuracy may be inferred once precision, linearity and specificity have been established.
准确度可以在精密度，线性及专属性建立之后推论得到；

4.2. Impurities (Quantitation) 杂质(定量)
Accuracy should be assessed on samples (drug substance/drug product) spiked with known amounts of impurities.
准确度可以通过向样品(原料药/制剂)中加入已知量杂质的方法来评价。
In cases where it is impossible to obtain samples of certain impurities and/or degradation products, it is considered acceptable to compare results obtained by an independent procedure (see 1.2.). The response factor of the drug substance can be used.
如果无法得到杂质和或降解产物的样品，可以通过与其它独立方法(见1.2节)的检测结果进行对比评估其准确度。可以使用原料药的响应因子。
It should be clear how the individual or total impurities are to be determined e.g., weight/weight or area percent, in all cases with respect to the major analyte.
需要说明单杂和总杂是如何测定的，如相对于主要被分析物所占的质量分数或面积百分比。
4.3. Recommended Data 可接受数据(数据要求)
Accuracy should be assessed using a minimum of 9 determinations over a minimum of 3 concentration levels covering the specified range (e.g., 3 concentrations/3 replicates each of the total analytical procedure).
准确度的评价需要在方法的线性范围内的三种浓度至少测定九次(按完整分析步骤对三种浓度每种浓度重复进样三次)。
Accuracy should be reported as percent recovery by the assay of known added amount of analyte in the sample or as the difference between the mean and the accepted true value together wit