APIC 201405原料药厂清洁验证指南：8.0残留量检测（上）（中英文）

2014-07-11 julia翻译 蒲公英

8.0 Determination of the Amount of Residue 残留量的检测

8.1 Introduction 介绍

This section provides a practical guidance for the determination of the amount of residue in cleaned equipment based on the requirements from regulatory authorities[1] and current guidelines on analytical validation.[2] Specific requirements for the validation of analytical and sampling methods for cleaning validation purposes are provided in this section, in addition to examples of sampling methods and the appropriate use of analytical methods.

本部分根据药监当局的要求和现行分析方法验证指南，提供了检测已清洁设备中残留物数量的实用指南。在本部分中给出了清洁验证中所用分析方法和取样方法的验证要求，以及取样方法和适当使用分析方法的例子。

The carryover acceptance limit (Mper) is a calculated figure that represents the specification limit for the equipment cleanliness (see Section 4.0, Acceptance Limits), however, the determination of the actual amount of residue (M) remaining in the equipment following cleaning must be achieved using appropriate methods i.e. for both the sampling method and the quantitation of the contaminant in the sample.

允许残留限度（Mper）是一个计算出的数值，代表设备清洁程度的质量标准限度（参见4.0部分，可接受限度），但是，对清洁后设备中残留物（M）实际数量的检测必须使用适当的方法来获得，即，针对取样方法和样品中污染物的定量。

Since the decision on the acceptable cleanliness of the equipment bears a potential risk to product quality, the method(s) used for the determination of M must be validated1 and the specificity, sensitivity and recovery of the method(s) should be determined as a minimum.

由于对设备可接受清洁度的决定会对产品质量有潜在风险，因此用于M检测的方法必须进行验证，必须至少验证方法的专属性、灵敏度和回收率。

8.2 Validation Requirements 验证要求

8.2.1 General 通则

The requirements for analytical method validation are defined in ICH Q2(R1), Validation of Analytical Procedures: Text and Methodology, November 2005. There are four types of analytical methods with principally different validation requirements; these are identification tests, tests for impurities (both quantitative and limit tests) and assay tests. The validation requirements for each method type are shown Table 1.

分析方法验证的要求在ICH Q2(R1)“分析方法验证：检验和方法学，2005年11月”中已给出。其中列出了四种不同类型的分析方法，各适用不同的验证要求，它们分别是：鉴别测试、杂质检验（定量和限度测试）和含量测试。每种类型方法的验证要求在表1中列出。

The list should be considered typical for the aforementioned analytical procedures, however, exceptions should be dealt with on a case-by-case basis. It should be noted that robustness is not listed in the table and should be considered at an appropriate stage in the development of the analytical procedure.

在上述的分析方法中必须考虑该清单，但是，要根据各案处理例外情况。要注意耐用性在表中并未列出，但在方法的研发过程中适当阶段需要进行考虑。

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.

在实际应用中，通常可以设计一个试验性工作，使得在方法设计时可以同步考虑方法的验证特性，例如，专属性、线性、范围、准确度和精密度。

The validation of an analytical method should occur in compliance with pre-established acceptance criteria that should be documented in a written general policy or Validation Plan. However, there should be one validation report per validated method that summarises the specific results.

分析方法验证必须符合预定的可接受限度。可接受限度应记录在书面的通用方针或验证中。但是，对每个验证过的方法必须有一个验证报告，总结验证结果。

TABLE 1 Requirement List for Analytical Validation

Characteristic	Type of Analytical Procedure
	Identification	Testing for Impurities		Assay
		Quantitative	Limit
Accuracy	—	+	—	+
Precision
Repeatability		+	—	+
Intermediate Precision	—	+1	—	+
Specificity2	+	+	+	+
Detection Limit	—	—3	+	+
Quantitation Limit	—	+	—	—
Linearity	—	+	—	+
Range	—	+	—	+
Key	—       Signifies that this characteristic is not normally evaluated. +   Signifies that this characteristic is normally evaluated. 1          In cases where reproducibility has been performed, intermediate precision is not needed. 2          Lack of specificity of one analytical procedure could be compensated by other supporting analytical procedure(s). 3          May be needed in some cases.

表1 方法验证需求清单

特性	分析方法类型
	鉴别	杂质检测		含量
		定量测试	限度测试
准确度	—	+	—	+
精密度
重复性		+	—	+
中间精密度	—	+1	—	+
专属性2	+	+	+	+
检测限	—	—3	+	+
定量限	—	+	—	—
线性	—	+	—	+
范围	—	+	—	+
说明	—       表示该特性一般不需要评估。 +   表示该特性一般要进行评估。 1          如果做了重复性测试，则不需要做中间精密度。 2          如果一个方法缺乏专属性，则可以用另一个支持性分析方法进行补偿。 3          有时需要。

The requirements for ‘Testing for Impurities’ are typically employed for the validation of analytical methods specific to cleaning validation.

在清洁验证专用分析方法验证中，一般应用“杂质检测”的要求。

The requirements for ‘Quantitative Testing for Impurities’ can apply, for example, in cases where a method should be suitable for several possible acceptance limits and therefore quantitation of the residue over a certain range may be necessary e.g. the measured amount of residue M must be compared with acceptance limits between 5 and 750 g/equipment. This is possible when the method will be used for several changeovers.

有时可以应用“杂质定量检测”要求，例如，如果一个方法适用于几个可能的可接受限度，因此可能需要对一定水平范围内的残留物进行定量检测，例如，残留物M的测得量必须与可接受标准5-750g/设备相比较。如果方法要用于几个产品更换情况下的检测，则可以使用上述方法。

The requirements for ‘Limit Testing for Impurities’ can apply, for example, in cases where the method should be suitable for one specific acceptance limit e.g. the measured M must be compared with Mper ≤ 105 g/equipment.

有时可以应用“杂质限度检测”要求，例如，如果方法需要符合一个特定的可接受限度，例如，对M的测量必须与Mper ≤ 105 g/设备进行比较。

8.2.2 Analytical Method Validation for Cleaning Validation 清洁验证所用分析方法验证

In the following sections, aspects of analytical method validation specific to cleaning validation are emphasised. For further details refer to ICH Q2 (R1).

在以下部分强调了清洁验证专用的分析方法要点。更详细的要求参见ICH Q2(R1)。

Specificity is a basic requirement for all analytical methods (see Table 1), however, in the case of cleaning validation it may occur, that not all potential impurities are clearly specified. It is important to note that in such a situation a specific method may not always detect all impurities. Studies should be performed to characterise the unknown impurities, develop and validate suitable analytical methods. However, this can be an unacceptably time consuming task. In this case a method that detects all potential impurities together can be suitable, even when it is not specific for each of the impurities. For example, in a situation where only non-volatile impurities occur, a dry residue determination method that is specific for the sum of non-volatile impurities could be used, provided that the validation requirements according to Table 1 are satisfied. In order to consider the equipment acceptable for use it must be assumed that the dry residue consists of the worst case impurity (most toxic, most active etc.). In some cases a combination of several methods can achieve the necessary specificity.

专属性是所有分析方法（见表1）都适用的基本要求，但是，在清洁验证中，可能并不是所有潜在杂质都能被明确，在这种情况下要重点注意即使是专属的方法，也并不一定能检测出所有杂质。可以研究未知杂质的特性，建立适当的检验方法并验证，但这样做所耗费的时间可能是无法接受的。这时，可能会适当的办法是采用一个能检出所有潜在杂质的方法，即使该方法对有些杂质不具有专属性。例如，如果只有不挥发性杂质产生，只要验证结果符合表1的要求，可以使用非专属性方法蒸发残渣来检测所有不挥发性杂质的总和。考虑到要使设备清洁程度可以接受，必须假定蒸发残渣结果中会包括最差情形的杂质（毒性最强、活性最强等）。有时，可以联合使用几个方法来达到必需的专属性要求。

After the completion of a cleaning validation study an unspecific method (e.g. dry residue) may be used for the routine verification of equipment cleaned by the validated cleaning procedure provided that it is shown that the unspecific method is suitable for the intended purpose. If possible, the sensitivity of impurity detection for cleaning validation should be determined for both the sampling and analytical methods together (see Section 7.2.4).

在清洁验证完成后，可以使用非专属性方法（例如，蒸发残渣）对按验证过的清洁程序清洁后的设备进行日常检测，只要该方法适合其既定用途。可能的情况下，清洁验证所用检验方法对杂质检测的灵敏度应与取样方法和分析方法一起测试（参见7.2.4部分）。

8.2.3 Detection and Quantitation Limits 定量限和检测限

Measured values below limit of quantification (LOQ) should be reported as the LOQ value (worst case approach). For example if the LOQ is 10 mg/l, the measured blank is 7 mg/l and the measured residue amount is 3 mg/l, the reported value for the sample should be equal to the LOQ i.e. 10 mg/l.

低于定量限（LOQ）应报告为定量限值（最差情况法）。例如，如果LOQ为10mg/L，测得空白结果为7mg/L，测得残留值为3mg/L，则该样品的残留值应报告为LOQ，即10mg/L。

Usually it can be assumed that, for quantitative impurity determination, the LOQ should approximately be 0.5 of the specification i.e. for cleaning validation 0.5 of the acceptance limit or lower. LOQ should never be higher than the acceptance limit. In the following sections three methods of LOQ/LOD determination are outlined:

一般可以假定，在杂质定量检测时，LOQ应为质量标准的约0.5倍，即对于清洁验证来说，可接受限度的0.5或更低。LOD不得高于可接受限度。在以下部分，列出了三种确定LOQ/LOD的方法：

? Based on Visual Evaluation 目视评估

Visual evaluation may be used for non-instrumental methods but may also be used with instrumental methods. Frequently this approach is used for TLC.

目视评估可以用于非仪器方法，但也可用于仪器分析方法。这种方法经常用于TLC。

? Based on Signal-to-Noise Approach 信噪比方法

This approach can only be applied to analytical procedures which exhibit baseline noise (e.g. GC, HPLC). A signal-to-noise ratio (S/N) between 3 or 2:1 is generally considered acceptable for estimating the detection limit (LOD) and a typical ratio for acceptable quantitation limit is 10:1 (LOQ). The value for S/N can be calculated according to Equation 1 and Figure 1:

本方法只能用于产生基线噪声的分析方法（例如，GC、HPLC）。信噪比（S/N）为2-3：1时，一般可以作为估计的检测限（LOD），10：1可以作为定量限（LOQ）。信噪比值可以根据公式1和图1计算：

                               
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where: H is the height of the peak from the mean baseline.

hn is the maximum deviation of the baseline within the range of 5 to 20 fold width of peak at half height.

其中：H为基线平均值的峰高

hn是基线在5-20部半峰高时峰宽范围内最大偏差

                               
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? Based on the Standard Deviation of the Response and the Slope

根据响应和斜率的标准偏差

The detection limit may be expressed by Equation 2 and the quantitation limit by Equation 3.

检查限可以采用公式2表示，定量限可以用公式3表示。

                               
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8.2.4 Determination of Recovery 回收率测定

If possible, the recovery of impurity detection for cleaning validation should be determined for the sampling and analytical methods together at least for recovery and sensitivity (Limit of Quantitation - LOQ, or Limit of Detection - LOD). This can be achieved, for example, by spiking a surface equivalent to the equipment surface (e.g. material, polish grade) with different known amounts of the impurity. The impurity can then be recovered and analysed using the same sampling and analytical methods that will be used for the cleaning validation study. The overall results from this procedure are then compared to criteria for detection or quantitation limits as defined in ICH Q2 (R1). Validation of the limits may be achieved by the analysis of samples known to be near at the limits.

可能的话，可以针对取样方法和检验方法同时进行清洁验证中杂质检测的回收率和灵敏度（定量限-LOQ，或检测限-LOD）。例如，可以采用与设备表面材质相同的表面材料（例如，材质、粗糙度），在上面加不同已知数量的杂质，然后采用与清洁验证研究相同的方法取样并分析。再将使用该方法所得的总体结果与ICH Q2(R1)中定义的检测限或定量限标准进行比较。可对接近限度的已知样品进行分析来验证分析方法的检测限和定量限。

The measured results are then compared to the actual amount applied to the surface. The recovery is typically determined during the accuracy determination and should be reported as a percentage of the known applied amount of the impurity.

将检测结果与实际数量进行比较，用于表面残留计算。回收率一般是在准确性测试中确定，应报告为已知杂质数量的百分比。

As an example, quantitative impurity determination recoveries of ≥ 90 % are usually regarded acceptable. For cleaning validation, recoveries of ≥ 90 % do not need to be taken into account for the calculation of the true value for M. Recoveries of < 90 % must be included in the calculation for M (see Equation 4) and recoveries of < 50 % should be omitted.

例如在杂质定量测试中，回收率≥ 90 %时一般认为是可以接受的。在清洁验证中，如果回收率≥ 90 %，在M残留量真值计算中可以不需要考虑回收率；如果回收率< 90 %，则需要在M计算时加以考虑（参见公式4），如果回收率<50%，则该方法不适用。

                               
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Where: M: True value for the amount of residue remaining in the equipment after cleaning;

Mres: The measured amount of residue (sampling and then analytical measurement);

R Recovery in % divided by 100 (e.g. for 75%, 75/100 = 0.75).

公式4：

其中M：清洁后设备上残留物的数量真值

Mres：测得残留量（取样后分析测量）

R：回收率除以100（例如，对于75%即为75/100=0.75）

请看下部分。