引 言

 

美国药典论坛USPPF43（2）中发布了关于残留溶剂药典方法确认和替代方法验证的新通则<1467>Residual Solvents- Verification of Compendial Procedures and Validation of Alternative Procedures。在这份通则中规定了在对残留溶剂方法进行确认和验证时包括的内容和可接受限度，对从事质量研究的人员具有指导意义。

 

前 言

 

通则<1467>规定USP<467>中1类、2类残留溶剂的方法学验证范围应在其浓度范围的50%~150%内，其中程序A和程序B中对照品溶液配制方法见USP<467>中Class 1 and Class 2 Residual Solvent项下。

 

USP<467>中规定的残留溶剂测定方法在实际使用时必须进行方法确认，并按新通则<1467>规定的方法进行确认。

 

替代方法（Alternative Procedures）包括与<467>中规定不同的方法或调整范围超出了USP<621>规定的方法。替代方法应经过充分验证，推荐的验证（限度测定或定量测定）规定详见本章Validation of Alternative Procedures项下规定，概要见表1。

 

Table1. Summary of Verification and Validation Requirements

表1. 方法确认和验证要求

 

Validation/Verification  Characteristics 验证/确认内容	Verification  of Compendial Procedures药典方法确认		Validation  of Alternative Procedures替代方法验证
	Limit  Test Methods 限度测定	Quantitative  Methods 定量测定	Limit  Test Methods 限度测定	Quantitative  Methods 定量测定
Specificity专属性	Yes	Yes	Yes	Yes
Detection  limit检测限	Yesa	No	Yes	No
Quantitation  limitb 定量限	No	Yesa	No	Yes
Accuracy准确度	No	Yes	No	Yes
Precision/repeatability精密度/重复性	No	Yes	No	Yes
Linearity线性	No	No	No	Yes
Range范围	No	Demonstrated  by accuracy and precision	No	Demonstrated  by accuracy and precision
Intermediate  precision中间精密度	No	No	No	Yes
Solution  stabilityc 溶液稳定性	Yes	Yes	Yes	Yes
Robustnessd 耐用性	No	No	Yese	Yese
a System  suitability may be used instead to demonstrate sensitivity. 系统适用性可代替灵敏度 b In  quantitative tests, quantitation limit may be demonstrated by accuracy  determination. 定量测定中，定量限可用准确度表示 c Solution  stability should be determined for the time line of the test. 溶液稳定性可通过测定的时间段来确定 d Evaluation  of robustness should be considered during the development phase and depends  on the type of procedure under study. 耐用性应在方法开发阶段考虑并与所选的方法类型有关 e The “Yes”  quotation here is intended to emphasize the importance of assessing the  robustness of the procedure before the implementation. “YES”用来强调在评估耐用性的重要性

 

Limit Procedures:  Procedure A and Procedure B

 

限度程序：程序A和程序B

 

The analytical characteristics to be verified include specificity, detectability, and solution stability.

 

确认项目应包括专属性、检测限、溶液稳定性。

 

VERIFICATION WHEN SOLVENTS LIKELY TO BE PRESENT (LTBP)ARE NOT KNOWN

 

可能存在的未知溶剂的确认

 

Samples样品

 

Prepare a reagent blank.配制空白溶剂

 

Prepare Standard solutions as described in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles.

 

对照品溶液（Standard solutions）按残留溶剂467中Class 1 and Class 2 Residual Solvents（1、2类残留溶剂）、Screening of Water-Soluble Articles（水溶性样品）和Screening of Water-Insoluble Articles（水不溶性样品）项下方法配制。

 

Prepare the Spiked sample solution as described for Class 1 System suitability solution in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles.

 

加标溶液（Spiked sample solution）按残留溶剂467中Class 1 and Class 2 Residual Solvents（1、2类残留溶剂）、Screening of Water-Soluble Articles（水溶性样品）和Screening of Water-Insoluble Articles（水不溶性样品）项下方法配制。

 

Specificity 专属性

 

Recommended acceptance criteria: Thereagent blank does not produce any significant interference with any of the peaks of the Standard solutions. The procedure must be able to separate acetonitrile and methylene chloride (Procedure A in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles), or acetonitrile and cis-dichloroethene (Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles) with are solution of NLT 1.0.

 

推荐接受标准：空白试剂不干扰对照品溶液中的任何色谱峰。乙腈与二氯甲烷的分离度（程序A中）或乙腈与顺式-1，2-氯乙烯（程序B中）的分离度NLT1.0。

 

Detection Limit (see Table 1,Footnote a) 检测限（见表1，脚注a）

 

Recommended acceptance criteria: The mean signal-to-noise ratio for each solvent in the Standard solution and Spiked sample solution (after correction for native solvent content) from at least three determinations from a single preparation is NLT 3.

 

推荐接受标准：对照品溶液和加标溶液中（根据实际含量校正）的每个溶剂的平均信噪比NLT3（至少测定3次）。

 

Solution Stability溶液稳定性

 

Recommended acceptance criteria: Detection limit should meet the requirements throughout the testing period.

 

推荐接受标准：整个测试过程中，检测限应符合要求。

 

VERIFICATION WHEN SOLVENTS LTBP ARE KNOWN

 

可能存在的已知溶剂的确认

 

Samples 样品

 

Prepare a reagent blank. 配制空白溶剂

 

Prepare Standard solution(s) and Sample solution(s) as described in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles depending on the specific sample solubility using only those solvents likely to be present (LTBP).

 

对照品溶液（Standard solutions）和样品溶液（Sample solution）根据可能存在溶剂的可能性按残留溶剂467中Class 1 and Class 2 Residual Solvents（1、2类残留溶剂）、Screening of Water-Soluble Articles（水溶性样品）和Screening of Water-Insoluble Articles（水不溶性样品）项下方法配制。

 

Prepare Spiked sample solution(s) asdescribed in Quantitative Procedures: Procedure C for solvents LTBP (corrected for native solvent content).

 

加标溶液（Spiked sample solution）按可能存在的溶剂（根据实际含量校正）定量测定程序C项下方法配制。

 

Specificity专属性

 

Recommended acceptance criteria: The reagent blank does not produce any significant interference with any of the peaks from solvents LTBP.

 

推荐接受标准：空白溶液不干扰可能存在溶剂的出峰。

 

The procedure must be able to separate each of the solvents in the Standard solution(s) from each other and from other peaks in the Spiked sample solution with a resolution of NLT 1.0. If the resolution between any pair of peaks is less than 1.5, then verification must demonstrate that the method is suitable for its intended use.

 

对照品溶液及加标溶液中每个峰的分离度应NLT1.0。如果两个峰的分离度小于1.5，应确认本方法能够符合预期的使用要求。

 

If the solvents present in the Standard solution(s) are not separated with a resolution of NLT 1.0 when using the Chromatographic System of Procedure A, then Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles should be used as confirmatory.

 

如果程序A中对照品溶液中分离度小于1.0，应使用程序B进行确认。

 

Detection Limit定量限

 

Recommended acceptance criteria: The mean signal-to-noise ratio for each solvent in the Standard solution and the Spiked sample solution (after correction for native solvent content) from at least three determinations from a single preparation is NLT 3.

 

推荐接受标准：对照溶液及加标溶液中（根据实际含量校正）每个溶剂峰的平均信噪比NLT3（至少测定3次）。

 

SolutionStability 溶液稳定性

 

Recommended acceptance criteria (not required if only running fresh solutions): Detection limit for limit tests and quantitation limit for quantitative tests should be met throughout the testing period.

 

推荐接受标准（如果临用新制，不需要考察）：在整个限度测定过程中，检测限应满足要求；在整个定量测定过程中，定量限应满足要求。

 

Quantitative Procedures: Procedure C 定量程序：程序C

 

[NOTE—When performing Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2Residual Solvents, 8.5 Quantitative Tests—Procedure C, solvents LTBP are typically known, either based on results from Procedure A or Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles or based on available knowledge. The analytical characteristics to be verified include specificity, accuracy (which addresses quantitation limit), range, repeatability, and solution stability.]

 

备注：当采用残留溶剂467中程序C时，确认项目应包括专属性，准确度（确定定量限），范围，重复性和溶液稳定性。

 

Samples样品

 

Preparea reagent blank.配制空白溶剂

 

Standard stock solution: Prepare a solution containing each solvent LTBP or each peak identified and verified by Procedure A and Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles, with a concentration asdescribed in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.5 Quantitative Tests—Procedure C for water-soluble articles or water-insoluble articles, as appropriate.

 

对照品储备溶液：配制包含可能存在的所有溶剂或根据程序A和程序B所确定的溶剂峰的溶液。方法见残留溶剂467中Class 1 and Class 2Residual Solvents（1、2类残留溶剂）、Quantitative Tests—Procedure C for water-soluble articles（水溶性样品） or water-insoluble articles（水不溶性样品）项下方法配制。

 

Sample stock solution: Prepare as described in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles. 

 

样品储备液：按残留溶剂467中Class 1 and Class 2 Residual Solvents（1、2类残留溶剂）、Screening of Water-Soluble Articles（水溶性样品）和Screening of Water-Insoluble Articles（水不溶性样品）项下方法配制。

 

Spiked sample solution: Prepare as described in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.5 Quantitative Tests—Procedure C, using the Standard stock solution. 

 

加标溶液：残留溶剂467中Class 1 and Class 2 Residual Solvents（1、2类残留溶剂）、Quantitative Tests—Procedure C使用对照品储备液配制。

 

Spiked sample solutions A, B, C, etc.: Prepare Spiked sample solutions with the sample matrix and spiked with each solvent LTBP or identified and verified by Procedure A and Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles, in triplicate, at NLT 3 levels covering the range of interest or at least 50%–150% of the corresponding concentration of the Standard solution as described for Procedure A in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening of Water-Insoluble Articles. [NOTE—The Spiked sample solution may be used as one of the solutions.]

 

加标溶液A,B,C等等：采用样品基质和可能存在的溶剂或根据程序A和程序B确定的溶液配制不少于3个浓度水平，至少覆盖50%~150%范围的溶液，每个浓度水平配制3份。

 

Specificity专属性

 

Recommended acceptance criteria: The procedure must be able to separate each of the solvents in the Standard solution(s) from each other and from other peaks inthe Spiked sample solution with a resolution of NLT 1.0. If the resolution between any pair of peaks is less than 1.5, then verification must demonstrate that the method is suitable for its intended use.

 

推荐接受标准：对照品溶液及加标溶液中各个峰的分离度应NLT1.0。如果两个峰的分离度小于1.5，应确认本方法能够符合预期的使用要求。

 

If the solvents present in the Standard solution(s) are not separated with a resolution of NLT 1.0, then chromatography described for Procedure B in Residual Solvents 〈467〉, 8. Analytical Procedures for Class 1 and Class 2 Residual Solvents, 8.2 Screening of Water-Soluble Articles or 8.3 Screening ofWater-Insoluble Articles should be used as confirmatory.

 

如果对照品溶液中存在分离度小于1.0的情况，应采用程序B方法进行确认。

 

Quantitation Limit定量限

 

Recommended acceptance criteria: The mean signal-to-noise ratio for each solvent in the Standard solution and the Spiked sample solution (after correction for native solvent content) from at least three determinations is NLT 10, or the Quantitation Limit may be demonstrated by Accuracy.

 

推荐接受标准：对照溶液及加标溶液中（根据实际含量校正）每个溶剂峰的平均信噪比NLT10（至少测定3次），或定量限可通过准确度证明。

 

Accuracy准确度

 

Recommended acceptance criteria: The mean recovery for Spiked sample solutions A, B, and C, when calculated relative to the Spiked sample solution, is 80%–120% of the expected the oretical amount. [NOTE—Recoveries should be corrected for native content of any solvent under test.]

 

推荐接受标准：加标溶液A,B,C相对于加标溶液的平均回收率应在理论含量的80%~120%范围内。（备注：各溶剂回收率应根据实际含量校正）

 

Precision/Repeatability精密度/重复性

 

Useat least six independent Spiked sample solution preparations from the same lot, prepared as described for the Spiked sample solution under the Accuracy test above, or use nine independent preparations as described for Spiked sample solutions A, B, C, etc.

 

采用同一批样品，按照准确度项下的规定配制至少6份加标溶液，或者配制9份加标溶液A,B,C.等等。

 

Recommended acceptance criteria: Relative standard deviation is NMT 20% for each solvent present.

 

推荐接受标准：各个存在溶剂的相对标准偏差应NMT20%。

 

Solution Stability: Demonstrate acceptable solution stability for the period of time to run the test.

 

溶液稳定性：根据溶液的测试时间来表示。

 

Recommended acceptance criteria: NMT 20% variation

 

推荐接受标准：变化率NMT20%。

 

VALIDATION OF ALTERNATIVE PROCEDURES

 

替代方法的验证

 

Chromatographic alternative procedures should meet the acceptance criteria for the analytical performance characteristics shown below. When non-chromatographic alternative procedures are validated, the analytical performance characteristics listed should be addressed, although, it may be appropriate to apply other analytical performance characteristics. For more information, refer to Validation of Compendial Procedures 〈1225〉.

 

色谱法可替代方法的验证应符合下文标准，非色谱替代方法被验证时，验证项目应该阐明，有关更多信息请参阅<1225>药典方法验证程序。

 

Limit Procedures限度测定

 

Analytical characteristics to be validated are specificity, detection limit, and solution stability. The same criteria used for Verification when solvents LTBP are known, as described above, may be used.

 

验证项目应包括专属性，检测限，溶液稳定性。接受标准同可能存在的已知溶剂的确认，见上文。

 

Quantitative Procedures定量测定

 

Analytical characteristics to be validated are specificity, linearity and range, quantitation limit, accuracy, repeatability, intermediate precision, and solution stability.

 

验证项目应包括专属性，线性，范围，定量限，准确度，重复性，中间精密度和溶液稳定性。

 

Samples样品

 

Preparea reagent blank.配制空白溶剂。

 

Spiked sample solutions P, Q, R, etc.: Prepare Spiked sample solutions with the sample matrix and spiked with each solvent LTBP at NLT 5 levels covering the range of interest. [NOTE—Results should be corrected for native content of any solvent under test.]

 

加标溶液P, Q, R等：采用样品基质和可能存在的溶剂配制不少于5个浓度水平，至少覆盖目标范围（备注：结果应根据实际含量校正）。

 

Specificity专属性

 

Recommended acceptance criteria: The analytical procedure must have the ability to assess unequivocally the analytes of interest in the presence of the components expected to be present.

 

推荐接受标准：分析程序必须有能力对预期存在的组分进行明确的评估。

 

Linearity and Range 线性和范围

 

Recommended acceptance criteria: Perform linear regression analysis on the results for Spiked sample solutions P, Q, R, etc. The coefficient of determination, r2, isNLT 0.90.

 

推荐接受标准：通过对加标溶液 P, Q, R等进行线性回归分析，测定的相关系数r2应NLT0.90.

 

Quantitation Limit 定量限

 

Recommended acceptance criteria: The mean signal-to-noise ratio for each solvent in the Standard solution and the Spiked sample solution(after correction for native solvent content) from at least three determinations is NLT 10. The Quantitation Limit may also be demonstrated by Accuracy.

 

推荐接受标准：标准溶液及加标溶液中（根据实际含量校正后）每个溶剂至少3次测定的信噪比的平均值不小于10。定量限可通过准确度证明。

 

Accuracy准确度

 

Recommended acceptance criteria: The mean recovery for each Spiked sample solution shouldbe 80%–120%. [NOTE—Recoveries should be corrected for native content of any solvent under test.]

 

推荐接受标准：每个加标溶液的平均回收率应在80%~120%范围内。（备注：各溶剂回收率应根据实际含量校正）

 

Repeatability重复性

 

Prepareat least six independent Spiked sample solution preparations from the same lot.

 

采用同一批样品，按照准确度项下的规定配制至少6份加标溶液。

 

Recommended acceptance criteria: Relative standard deviation is NMT 20% for each solvent present.

 

推荐接受标准：各个存在溶剂的相对标准偏差应NMT20%。

 

Intermediate Precision中间精密度

 

Perform the Repeatability test over at least two independent events, for example on different days, and/or using different instruments and/or analysts.

 

至少两个独立不同情况下进行另一份重复性测试，例如不同天，和/或不同仪器，和/或不同分析员。

 

Recommended acceptance criteria: Testing for intermediate precision should demonstrate that the method is suitable for its intended use.

 

推荐接受标准：中间精密度测试应表明方法能够达到预期的使用目的。

 

Solution Stability: Standard solutions and Spiked sample solutions should be stable throughout the testing period.

 

溶液稳定性：整个测试过程中，对照品溶液和加标溶液应稳定。

 

Recommended acceptance criteria: NMT 20% variation (USP41)

 

推荐接受标准：变化率NMT20%（USP41）。