接上部分。
8.2.5 Validation Requirements for Quantitative Testing of Impurities 杂质定量测试的验证要求
The requirements for the validation of quantitative testing of impurities according to ICH Q2 (R1) are shown in Table 2, including proposed acceptance criteria (as an example only). Alternative acceptance criteria may be established based on sound scientific rationale.
根据ICH Q2(R1)，杂质定量检测方法的验证要求在表2中列出，包括制订的可接受标准（只是举例）。可以根据科学合理的原则制订适当的可接受标准。
It is important to note, that the summarised requirements should be used for the validation of quantitative testing for impurities during cleaning validation studies. Validation of quantitative testing for impurities is usually applied when the analytical method will be used for several specifications of the residue amount in the equipment.
重点要注意的是，在清洁验证研究中，定量检测方法验证应满足所有要求。如果分析方法将用于设备中残留量有不同的几个质量标准，则一般采用杂质定量方法验证要求。
The lowest foreseen acceptance limit is referred to as MperMin and the highest limit as MperMax in Table 2. For only one specific acceptance limit normally limit testing for impurities and the corresponding validation of the analytical method is sufficient. If the validation of quantitative testing for impurities will be used for one specific acceptance limit, then MperMin = MperMax = Mper.
最低预期可接受限度，在表2中称为MperMin，和最高限度，称为MperMax。对于只有一个特定的可接受限度，一般可以使用杂质限度测试方法，对方法只要做相应的验证即可。如果要将杂质定量方法的验证用于单一可接受限度，则MperMin = MperMax = Mper。
For the experimental work described in Table 2, the samples can be spiked with appropriate levels of the impurities (when standards are available) or compared with another well-characterised procedure (when standards are not available) to obtain the true value of the analyte concentration.
表2中试验里，可以采用杂质（如果可以获得标准品的话）加入样品中至适当的浓度水平，或与另一个经过确认的检验方法进行对比（如果不能得到标准品的话），以得到被分析物的真实浓度值。
TABLE 2 Validation Requirements
Experiments | Possible Acceptance Criteria |
Accuracy: |
Perform a minimum of 9 determinations over a minimum of 3 concentration levels covering the specific range (e.g. 3 concentrations/3 replicates each of the total analytical procedure). Determine analyte with respect to the total amount of residue in the sample (e.g. weight/weight). Report: |
□ Accuracy as percent recovery or | 90.00-110.00% |
□ Difference between the mean and the accepted true value. | ≤10.00% (P=95%) |
□ Confidence intervals. |
|
Precision: |
Investigate using homogenous, authentic samples or (if not possible) artificially prepared sample. Perform a minimum of 9 determinations covering the specified range for the procedure (e.g. 3 concentrations/3 replicates each) or a minimum of 6 determinations at 100% of the test concentration. |
Repeatability (intra-assay precision): |
Establish precision under the same operating conditions over a short interval of time. Report: |
□ Standard deviation (interdependent with S) | See S |
□ Overall relative standard deviation over the whole range of the method | ≤10.00% |
□ Relative standard deviation within one concentration level | ≤20.00% |
□ Confidence interval |
|
Intermediate Precision (may include robustness, ruggedness): |
establish precision on different days, for different analysts, on different equipment and after variation of method parameters (= robustness, e.g. stability of solutions, variations of pH, of mobile phase composition, of flow rate, of temperature, of columns etc.). It is not necessary to study these effects individually. Experimental design (matrix) may be applied. Report: |
□ Standard deviation (interdependent with relative standard deviation) | See S |
□ Relative standard deviation | 3×S from repeatability or 10% whichever is greater |
□ Confidence interval |
|
Specificity: |
Demonstrate the discrimination of analyte in the presence of the other impurities: |
□ Test samples containing the analyte and other impurities. Obtain positive and correct results for the analyte. | Specify acceptable deviation |
□ Test samples without the analyte. | Negative results |
□ For chromatographic procedures use representative chromatograms to document specificity. Label individual components appropriately. | Specify acceptable resolution of peaks |
Linearity: |
Measure a minimum of 5 concentrations across the range of the procedure (dilute standard stock solution or prepared synthetic mixtures). Plot the signals as function of concentration. Evaluate the plot: |
□ Visually | Linear |
□ Statistically (e.g. regression line by the method of least squares) |
|
Correlation coefficient | ≥0.99000 |
y-intercept | Confidence band (P = 95%) contains 0 |
Slope of the regression line |
|
Residual sum of squares |
|
Range: |
Confirm that the analytical procedure provides an acceptable degree of linearity, accuracy and precision within or at the extremes of the specified range. Minimum specified ranges: |
□ From the reporting level to 120% of MperMax. The reporting level for cleaning validation reasonably will be the LOQ. However, the reporting level must be below MperMin and should be below or at 80% of MperMin. | From LOQ or 80% of MperMin to 120% of MperMax. |
表2 验证要求
测试项目 | 可能的可接受标准 |
准确度 |
在指定的浓度范围内对3个浓度进行至少9次检测（例如，3个浓度各按完整的分析方法检测3次）。测试残留物在样品中的总量（例如重量/重量）。 报告： |
□精密度，表达为回收率百分比或 | 90.00-110.00% |
□平均值与所接受的真值之间的差异. | ≤10.00% (P=95%) |
□置信区间 |
|
精密度: |
使用均匀的具有标准值的样品或（如果没有标准样品的话）人为制备的样品，在指定范围内采用指定方法（例如，3个浓度各3次重复检测）检测至少9次，或在检测浓度100%水平测试至少6次。 |
重复性（含量精密度） |
建立相同操作条件下较短时间内的精密度 报告：: |
□标准偏差（与S相互依存） | 参见 S |
□整个方法浓度范围内的总体相对标准偏差 | ≤10.00% |
□一个浓度水平的相对标准偏差 | ≤20.00% |
□置信区间 |
|
中间精密度（可以包括耐用性） |
在不同天、不同化验员采用不同设备对方法参数进行变动（=耐用性，例如，溶液稳定性、pH值变化、流动相比例、流速、温度、柱子等）检测方法精密度。不需要单独研究各因素的影响，可以使用试验设计（矩阵）。 报告： |
□标准偏差（依赖于相对标准偏差） | 参见S |
□相对标准偏差 | 重复性中的S值乘3或10%，取大者 |
□置信区间 |
|
专属性 |
证明被分析物与其它杂质的分离度 |
□检验含有被测试物和杂质的样品，得到被分析物的符合的正确结果。 | 指定可接受偏差 |
□检测不含被分析物的样品。 | 测试结果不含被分析物 |
□对于色谱方法，采用代表性图谱记录专属性。对各化合物进行恰当标识。 | 指定各峰间的可接受分离度 |
线性 |
检验方法范围内最少5个浓度检测（稀释标准贮备液或制备的合成混合物）。画出浓度-信号点，评估： |
□目视 | 线性 |
□统计数据（例如，最小平方法回归线） |
|
相关系数 | ≥0.99000 |
Y轴截距 | 置信区间 (P = 95%) contains 0 |
回归线斜率 |
|
残差平方和 |
|
范围： |
确认检验方法的线性可以接受，精密度和准确度在指定的范围内或在范围极值。最小指定范围： |
□ 报告限至最大样品浓度的120%。合理的清洁验证的报告水平应在定量限，但是报告水平必须低于最小样品浓度，低于或等于最小样品浓度的80%。 | 从定量限或最小浓度的80%到最大浓度的120%。 |
8.3 Sampling Methods 取样方法
In order to demonstrate that the plant equipment is verified clean and meets the pre-defined acceptance criteria, sampling and analysis should be carried out using the methods described in the following sections. Justification should be provided for the selection of the appropriate verification technique on a case by case basis. A combination of the two methods is generally the most desirable. For all methods the sampling points should be fixed in a manner such that the true contamination of the equipment will be reflected.
为了证明工厂设备是清洁的，且符合预定的可接受标准，应采用以下部分所述的方法进行取样和分析。应各案论述对选择技术的恰当性。一般最好采用同时采用两种取样方法。对所取样方法，取样点应以一定方式固定，以反映设备中的真实污染情况。
8.3.1 Swab sampling (Direct Surface Sampling) 擦拭取样（表面直接取样）
Swab sampling of the direct surface is designed to test small sections of the equipment surface for the presence of residues. Samples should be taken from all main equipment items and since swab sampling does not cover the entire equipment surface area, justification should be provided for the choice of the area for swabbing.
对设备表面进行直接擦拭取样设计用于检测设备上小面积上残留物。样品应从所有主要设备上取样，由于擦拭取样并不覆盖整个设备表面，因此要对于取样部位的选择做出论述。
Typically a small area of the cleaned equipment is swabbed with a material according to a pre-defined method i.e. swab material, solvent and technique. The swab sample can then be extracted and examined using a suitable analytical method.
一般来说，对已清洁设备的一个较小面积采用预定的方法，例如擦拭材料、溶剂和方法，进行擦拭取样。对擦拭样品进行提取，采用适当的分析方法进行检测。
The quantified residue obtained from the sample is then extrapolated to the whole equipment (see Equation 6).
将样品测试所得的残留量值外推至整个设备（参见公式6）。
It is important: 以下内容应重视
l That the validation of the swab sampling is performed on the same surface (material, polish grade, area in dm2) and with the same materials as the routine sampling of the equipment.
l 擦拭取样方法验证应采用相同表面（材质、光洁度、面积以dm2为单位），以及设备常规取样时相同的取样棒
l That the choice of swabbing material considers extractable materials that could interfere with the expected residue.
l 选择擦拭材料时，要考虑可萃出物料可能对预期的残留物产生影响
l To ensure that the sampling points represent the worst case areas of the equipment.
l 要保证取样点代表设备的最差面积
The disadvantage of this sampling method for often complex API equipment is that difficult to reach areas (e.g. sealings, condensers, transfer pipework) may not be accessible by swabbing. Nevertheless these areas may be the critical areas for the determination of the amount of residue in the equipment.
该取样方法的缺点是有些原料药设备比较复杂，有些部分难以取到擦拭样品（例如，密封处、冷凝器、转移用管道），而这些面积可能对于设备残留量的测定比较关键。
M | Amount of residue in the cleaned equipment in mg. | 已清洁设备中残留量mg |
WF | Recovery rate for the whole chain swab/analytical method (e.g. 0.8 for 80%). | 整个设备链的擦拭/分析方法回收率（例如0.8代表80%） |
Ftot | The entire inner surface of the equipment in dm2 | 设备整个内表面积，平方分米 |
Mi | Amount of residue (e.g. previous product) in the sample i in mg. | 样品中的残留量（例如，上一产品）mg |
Ci | Gross amount of residue in the sample i in mg. | 样品 i 中残留总量mg |
CBi | Blank of the sample i in mg. To establish the blank, a swab (or several swabs) can be treated in the similar way as a sampling swab except swabbing of the contaminated surface. Usually one and the same blank can be used for all N sampling swabs. | 空白样品 i 重量mg 为建立空白，可以将一个（或几个）取样用空白棉签采用与样品同等方法处理。一般一个相同的空白可以用于所有N个取样棉签。 |
Fi | Area swabbed by the swab i in dm2. | 擦拭取样的面积，平方分米 |
N | Number of swab samples. | 样品数量 |
i | Sample identifier (current number from 1 to N). | 样品识别号（现在编号为1至N） |
The first production batch of the following product may be sampled and analysed for impurities (for preceding product) since chromatographic analytical methods will typically be used (e.g. HPLC, GC, TLC).
由于现在普遍使用色谱分析方法（例如HPLC、GC、TLC），可以对下一产品的第一个生产批号取样，分析其中（上一产品）杂质。
8.3.2 Rinse or Wash Solvent Sampling 淋洗或冲洗取样
In cases where swabbing is not possible, for example restricted access, swabbing may be substituted by the analysis of final rinse solutions. Rinse samples can be used to determine the carryover of residues over a large surface area and cover all main process items including transfer pipework. In cases where swab sampling is not practical, it is acceptable to analyse only rinse samples, however this should be justified as part of the validation study.
如果没办法进行擦拭取样，例如不能入口受限，可以对最终淋洗液进行检测来替代擦拭样品。淋洗样品可以用于确定一个很大表面积上的残留量，并覆盖所有主要工艺设备，包括物料管道。如果擦拭取样不实际，可以接受只对淋洗样品进行检测，但应作为验证研究的一部分进行论述。
This section outlines the quantitation of the amount of residue remaining in the equipment after cleaning based on the amount of residue in the last rinse of the routinely used cleaning procedure.
本章列出了根据清洁程序中最后一次淋洗液中残留量，计算清洁后设备中残留量的定量方法。
The residue amount in the equipment can be assumed to be equal to the amount of residue in the last wash or rinse solvent portion. The assumption is based on the worst case consideration that a further rinse (or any reaction) would not remove more than the same amount of residue present in the analysed rinse sample.
可以假定设备中残留量等于最后冲洗或淋洗溶剂中的总量。该假设是基于最差情形考虑，即进一步淋洗（或任何反应）将无法得到比分析用淋洗样品结果更高的残留物。
The advantage of the rinse sampling method is the whole equipment will be reached by the solvent, including difficult to reach locations that cannot be disassembled. Therefore, if appropriately designed, this method will give the best indication of the amount of residue remaining in the equipment.
淋洗取样方法的优点是整个设备都会被溶剂洗到，包括最难触及且不能拆卸的部位。因此，如果对淋洗方法进行了适当的设计，该方法能最好地显示设备中残留量。
For quantitation, a solvent sample (e.g. 1 litre) is removed and the residue in the sample is determined by a suitable analytical method, which can then be extrapolated to the whole equipment according to Equation 5.
在定量检测时，取一个溶剂的样品（例如1L），对样品中的残留采用适当的分析方法进行检测，然后根据公式5外推至整个设备。
Equation 5: M = V × (C – CB)
Where 其中
M | Amount of residue in the cleaned equipment in mg. | 清洁后的设备中的残留量mg |
V | Volume of the last rinse or wash solvent portion in litres. | 最后淋洗或冲洗溶剂体积，L |
C | Concentration of impurities in the sample in mg/l. | 样品中杂质浓度，mg/l |
CB | Blank of the cleaning or rinsing solvent in mg/l. If several samples are taken during one run, one and the same blank can be used for all samples provided the same solvent lot was used for the whole run. | 清洁或淋洗溶剂空白水平，mg/l 如果在一个轮次中取了几个样品，并且在整个轮次中使用的溶剂是同一个批号，则可以在所有样品计算中使用同一个空白。 |
8.3.3 Stamps 邮戳法
In this exceptionally used sampling method, “coins” (or stamps) are placed on appropriate sampling points in the equipment during the manufacture of the previous product and during cleaning. After cleaning, the contamination on the coins can be analysed and the overall contamination can be calculated by extrapolation to the whole equipment. For quantitation, the coins may be firstly swabbed followed by further analysis of the samples.
这是一种在例外情况下使用的取样方法。取一个“圆片”（或邮票），置于设备中适当的取样点上，进行上一产品的生产及清洁。在清洁后，对硬币上的污染物进行检测，采用外推法计算整个设备的全部污染。如果是要进行定量分析，则对圆片进行擦拭取样，然后对样品进行进一步分析。
8.4 Analytical Methods 分析方法
A sample isolated by either of the sampling methods discussed in Section 8.3 should be analysed by a suitable analytical method (e.g. HPLC, GC, GC-MS, TLC, dry residue, TOC, UV, titration, conductivity or pH). The suitability of the method can be documented by appropriate validation as detailed in Section 8.2.
按第8.3部分所讨论的取样方法之一所获得的样品应采用适当的分析方法（例如，HPLC、GC、GC-MS、TLC、蒸发残渣、TOC、UV、滴定、电导率或pH值）进行检测。方法的适用性可以依据8.2部分指导进行适当的验证并记录。
A combination of analytical methods can be used if appropriate. For example evaporation of the solvent sample and analysis of the dry residue by another method (e.g. HPLC) can enhance the sensitivity of the final analytical method by a factor 106. Alternatively, the use of several methods (e.g. titration, HPLC) can provide the required specificity.
适当时，可以合并使用几个分析方法。例如，将溶剂样品蒸发，采用另一个方法（例如HPLC）分析残渣。这样可以增加最后分析方法的灵敏度到100万倍；而采用几个方法（例如，滴定、HPLC）可以达到所需的专属性。
[2]ICH Q2 (R1), Validation of Analytical Procedures: Text and Methodology, November 2005
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