引 言

 

美國藥典論壇USPPF43（2）中發(fā)布了關(guān)于殘留溶劑藥典方法確認和替代方法驗證的新通則<1467>Residual Solvents- Verification of Compendial Procedures and Validation of Alternative Procedures。在這份通則中規(guī)定了在對殘留溶劑方法進行確認和驗證時包括的內(nèi)容和可接受限度，對從事質(zhì)量研究的人員具有指導(dǎo)意義。

 

前 言

 

通則<1467>規(guī)定USP<467>中1類、2類殘留溶劑的方法學(xué)驗證范圍應(yīng)在其濃度范圍的50%~150%內(nèi)，其中程序A和程序B中對照品溶液配制方法見USP<467>中Class 1 and Class 2 Residual Solvent項下。

 

USP<467>中規(guī)定的殘留溶劑測定方法在實際使用時必須進行方法確認，并按新通則<1467>規(guī)定的方法進行確認。

 

替代方法（Alternative Procedures）包括與<467>中規(guī)定不同的方法或調(diào)整范圍超出了USP<621>規(guī)定的方法。替代方法應(yīng)經(jīng)過充分驗證，推薦的驗證（限度測定或定量測定）規(guī)定詳見本章Validation of Alternative Procedures項下規(guī)定，概要見表1。

 

Table1. Summary of Verification and Validation Requirements

表1. 方法確認和驗證要求

 

Validation/Verification  Characteristics 驗證/確認內(nèi)容	Verification  of Compendial Procedures藥典方法確認		Validation  of Alternative Procedures替代方法驗證
	Limit  Test Methods 限度測定	Quantitative  Methods 定量測定	Limit  Test Methods 限度測定	Quantitative  Methods 定量測定
Specificity專屬性	Yes	Yes	Yes	Yes
Detection  limit檢測限	Yesa	No	Yes	No
Quantitation  limitb 定量限	No	Yesa	No	Yes
Accuracy準確度	No	Yes	No	Yes
Precision/repeatability精密度/重復(fù)性	No	Yes	No	Yes
Linearity線性	No	No	No	Yes
Range范圍	No	Demonstrated  by accuracy and precision	No	Demonstrated  by accuracy and precision
Intermediate  precision中間精密度	No	No	No	Yes
Solution  stabilityc 溶液穩(wěn)定性	Yes	Yes	Yes	Yes
Robustnessd 耐用性	No	No	Yese	Yese
a System  suitability may be used instead to demonstrate sensitivity. 系統(tǒng)適用性可代替靈敏度 b In  quantitative tests, quantitation limit may be demonstrated by accuracy  determination. 定量測定中，定量限可用準確度表示 c Solution  stability should be determined for the time line of the test. 溶液穩(wěn)定性可通過測定的時間段來確定 d Evaluation  of robustness should be considered during the development phase and depends  on the type of procedure under study. 耐用性應(yīng)在方法開發(fā)階段考慮并與所選的方法類型有關(guān) e The “Yes”  quotation here is intended to emphasize the importance of assessing the  robustness of the procedure before the implementation. “YES”用來強調(diào)在評估耐用性的重要性

 

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.

 

確認項目應(yīng)包括專屬性、檢測限、溶液穩(wěn)定性。

 

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

 

可能存在的未知溶劑的確認

 

Samples樣品

 

Prepare a reagent blank.配制空白溶劑

 

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.

 

對照品溶液（Standard solutions）按殘留溶劑467中Class 1 and Class 2 Residual Solvents（1、2類殘留溶劑）、Screening of Water-Soluble Articles（水溶性樣品）和Screening of Water-Insoluble Articles（水不溶性樣品）項下方法配制。

 

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.

 

加標溶液（Spiked sample solution）按殘留溶劑467中Class 1 and Class 2 Residual Solvents（1、2類殘留溶劑）、Screening of Water-Soluble Articles（水溶性樣品）和Screening of Water-Insoluble Articles（水不溶性樣品）項下方法配制。

 

Specificity 專屬性

 

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.

 

推薦接受標準：空白試劑不干擾對照品溶液中的任何色譜峰。乙腈與二氯甲烷的分離度（程序A中）或乙腈與順式-1，2-氯乙烯（程序B中）的分離度NLT1.0。

 

Detection Limit (see Table 1,Footnote a) 檢測限（見表1，腳注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.

 

推薦接受標準：對照品溶液和加標溶液中（根據(jù)實際含量校正）的每個溶劑的平均信噪比NLT3（至少測定3次）。

 

Solution Stability溶液穩(wěn)定性

 

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

 

推薦接受標準：整個測試過程中，檢測限應(yīng)符合要求。

 

VERIFICATION WHEN SOLVENTS LTBP ARE KNOWN

 

可能存在的已知溶劑的確認

 

Samples 樣品

 

Prepare a reagent blank. 配制空白溶劑

 

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).

 

對照品溶液（Standard solutions）和樣品溶液（Sample solution）根據(jù)可能存在溶劑的可能性按殘留溶劑467中Class 1 and Class 2 Residual Solvents（1、2類殘留溶劑）、Screening of Water-Soluble Articles（水溶性樣品）和Screening of Water-Insoluble Articles（水不溶性樣品）項下方法配制。

 

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

 

加標溶液（Spiked sample solution）按可能存在的溶劑（根據(jù)實際含量校正）定量測定程序C項下方法配制。

 

Specificity專屬性

 

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

 

推薦接受標準：空白溶液不干擾可能存在溶劑的出峰。

 

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.

 

對照品溶液及加標溶液中每個峰的分離度應(yīng)NLT1.0。如果兩個峰的分離度小于1.5，應(yīng)確認本方法能夠符合預(yù)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中對照品溶液中分離度小于1.0，應(yīng)使用程序B進行確認。

 

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.

 

推薦接受標準：對照溶液及加標溶液中（根據(jù)實際含量校正）每個溶劑峰的平均信噪比NLT3（至少測定3次）。

 

SolutionStability 溶液穩(wěn)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.

 

推薦接受標準（如果臨用新制，不需要考察）：在整個限度測定過程中，檢測限應(yīng)滿足要求；在整個定量測定過程中，定量限應(yīng)滿足要求。

 

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.]

 

備注：當采用殘留溶劑467中程序C時，確認項目應(yīng)包括專屬性，準確度（確定定量限），范圍，重復(fù)性和溶液穩(wěn)定性。

 

Samples樣品

 

Preparea reagent blank.配制空白溶劑

 

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.

 

對照品儲備溶液：配制包含可能存在的所有溶劑或根據(jù)程序A和程序B所確定的溶劑峰的溶液。方法見殘留溶劑467中Class 1 and Class 2Residual Solvents（1、2類殘留溶劑）、Quantitative Tests—Procedure C for water-soluble articles（水溶性樣品） or water-insoluble articles（水不溶性樣品）項下方法配制。

 

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. 

 

樣品儲備液：按殘留溶劑467中Class 1 and Class 2 Residual Solvents（1、2類殘留溶劑）、Screening of Water-Soluble Articles（水溶性樣品）和Screening of Water-Insoluble Articles（水不溶性樣品）項下方法配制。

 

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. 

 

加標溶液：殘留溶劑467中Class 1 and Class 2 Residual Solvents（1、2類殘留溶劑）、Quantitative Tests—Procedure C使用對照品儲備液配制。

 

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.]

 

加標溶液A,B,C等等：采用樣品基質(zhì)和可能存在的溶劑或根據(jù)程序A和程序B確定的溶液配制不少于3個濃度水平，至少覆蓋50%~150%范圍的溶液，每個濃度水平配制3份。

 

Specificity專屬性

 

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.

 

推薦接受標準：對照品溶液及加標溶液中各個峰的分離度應(yīng)NLT1.0。如果兩個峰的分離度小于1.5，應(yīng)確認本方法能夠符合預(yù)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.

 

如果對照品溶液中存在分離度小于1.0的情況，應(yīng)采用程序B方法進行確認。

 

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.

 

推薦接受標準：對照溶液及加標溶液中（根據(jù)實際含量校正）每個溶劑峰的平均信噪比NLT10（至少測定3次），或定量限可通過準確度證明。

 

Accuracy準確度

 

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.]

 

推薦接受標準：加標溶液A,B,C相對于加標溶液的平均回收率應(yīng)在理論含量的80%~120%范圍內(nèi)。（備注：各溶劑回收率應(yīng)根據(jù)實際含量校正）

 

Precision/Repeatability精密度/重復(fù)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.

 

采用同一批樣品，按照準確度項下的規(guī)定配制至少6份加標溶液，或者配制9份加標溶液A,B,C.等等。

 

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

 

推薦接受標準：各個存在溶劑的相對標準偏差應(yīng)NMT20%。

 

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

 

溶液穩(wěn)定性：根據(jù)溶液的測試時間來表示。

 

Recommended acceptance criteria: NMT 20% variation

 

推薦接受標準：變化率NMT20%。

 

VALIDATION OF ALTERNATIVE PROCEDURES

 

替代方法的驗證

 

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?.

 

色譜法可替代方法的驗證應(yīng)符合下文標準，非色譜替代方法被驗證時，驗證項目應(yīng)該闡明，有關(guān)更多信息請參閱<1225>藥典方法驗證程序。

 

Limit Procedures限度測定

 

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.

 

驗證項目應(yīng)包括專屬性，檢測限，溶液穩(wěn)定性。接受標準同可能存在的已知溶劑的確認，見上文。

 

Quantitative Procedures定量測定

 

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

 

驗證項目應(yīng)包括專屬性，線性，范圍，定量限，準確度，重復(fù)性，中間精密度和溶液穩(wěn)定性。

 

Samples樣品

 

Preparea reagent blank.配制空白溶劑。

 

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.]

 

加標溶液P, Q, R等：采用樣品基質(zhì)和可能存在的溶劑配制不少于5個濃度水平，至少覆蓋目標范圍（備注：結(jié)果應(yīng)根據(jù)實際含量校正）。

 

Specificity專屬性

 

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.

 

推薦接受標準：分析程序必須有能力對預(yù)期存在的組分進行明確的評估。

 

Linearity and Range 線性和范圍

 

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.

 

推薦接受標準：通過對加標溶液 P, Q, R等進行線性回歸分析，測定的相關(guān)系數(shù)r2應(yīng)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.

 

推薦接受標準：標準溶液及加標溶液中（根據(jù)實際含量校正后）每個溶劑至少3次測定的信噪比的平均值不小于10。定量限可通過準確度證明。

 

Accuracy準確度

 

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.]

 

推薦接受標準：每個加標溶液的平均回收率應(yīng)在80%~120%范圍內(nèi)。（備注：各溶劑回收率應(yīng)根據(jù)實際含量校正）

 

Repeatability重復(fù)性

 

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

 

采用同一批樣品，按照準確度項下的規(guī)定配制至少6份加標溶液。

 

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

 

推薦接受標準：各個存在溶劑的相對標準偏差應(yīng)NMT20%。

 

Intermediate Precision中間精密度

 

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

 

至少兩個獨立不同情況下進行另一份重復(fù)性測試，例如不同天，和/或不同儀器，和/或不同分析員。

 

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

 

推薦接受標準：中間精密度測試應(yīng)表明方法能夠達到預(yù)期的使用目的。

 

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

 

溶液穩(wěn)定性：整個測試過程中，對照品溶液和加標溶液應(yīng)穩(wěn)定。

 

Recommended acceptance criteria: NMT 20% variation (USP41)

 

推薦接受標準：變化率NMT20%（USP41）。