目的:基于不确定度曲线建立丹参酮提取物中丹参酮ⅡA含量的近红外(NIR)定量分析方法。方法:采集丹参酮提取物近红外光谱,采用HPLC作为参考方法分析丹参酮提取物中丹参酮ⅡA的含量,建立丹参酮提取物中ⅡA含量和近红外光谱之间的偏最小二乘(PLS)定量模型。通过K-S(Kennard-Stone)方法划分校正集和校正测试集,比较不同的光谱预处理方法建立最优PLS定量回归模型。采用“6×3×3”验证实验得到验证数据,得出NIR分析方法的验证信息,并计算NIR定量分析方法的不确定度,构建不确定度曲线并评价所建立方法的有效性。结果:选择1std数据预处理方法来建立PLS定量回归模型,所建立的NIR定量分析方法真实性,精密度良好,丹参酮ⅡA含量的有效定量范围为2.04%~27.64%,分析结果准确有效。结论:不确定曲线策略可以同时提供NIR分析方法的验证信息和不确定度信息,能够降低分析方法的使用风险,保障NIR分析结果的准确性和可靠性。
Objective: To establish a near infrared (NIR) quantitative analysis method for testing tanshinone ⅡA content in tanshinone extract based on uncertainty profile. Methods: The near-infrared spectrum of tanshinone extract was collected, and the content of tanshinone ⅡA in tanshinone extract was analyzed by HPLC as a reference method. Partial least squares (PLS) quantitative model between the content of tanshinone ⅡA in tanshinone extract and the near-infrared spectrum was established. The calibration set and calibration test set were divided by K-S (Kennard-Stone) method, and the optimal PLS quantitative regression model was established by comparing different spectral pretreatment methods. The validation information of the NIR analysis method was obtained by the “6×3×3” validation experiments, the uncertainty of the NIR quantitative analysis method was calculated, the uncertainty profile was constructed to evaluate the effectiveness of the NIR method. Results: The quantitative regression model of PLS was established after the 1std data pretreatment method. The trueness and precision of the established NIR quantitative analysis method was good. The effective quantitative range of tanshinone ⅡA content was 2.04%-27.64%, and the analysis results were accurate and effective. Conclusion: The uncertainty profile strategy can provide the validation information and uncertainty information of NIR analysis method at the same time, reduce the risk of using the analysis method, and ensure the accuracy and reliability of NIR analysis results.
[1] Pharmaceutical Analytical Sciences Group (PASG). NIR Sub-Group Guidelines for the Development and Validation of Near-infrared Spectroscopic Methods in the Pharmaceutical Industry[S]. 2002
[2] GINOT YM, BERNARD-MOULIN P, CINIER R, et al. Quantitative analysis using near infrared spectroscopy, Application to tablet content uniformity, I. Methodology [J]. STP Pharm Pratiques, 2010, 20(2):117
[3] The European Agency for the Evaluation of Medicinal Products, Note for guidance on the use of near-infrared spectroscopy by the pharmaceutical industry and the data requirements for news submissions and variations[S]. 2003
[4] The European Agency for the Evaluation of Medicinal Products. Guideline on the Use of Near-Infrared Spectroscopy by the Pharmaceutical Industry and the Data Requirements for News Submissions and Variations, EMEA/CHMP/CVMP/QWP/17760/2009 Rev 2[S]. 2009
[5] BLANCO M, PEGUERO A. Analysis of pharmaceuticals by NIR spectroscopy without a reference method[J]. TrAC-Trends Anal Chem, 2010, 29(10):1127
[6] CORREDOR CC, BU D, BOTH D. Comparison of near infrared and microwave resonance sensors for at-line moisture determination in powders and tablets-science direct[J]. Anal Chim Acta, 2011, 696(1-2):84
[7] LIEW CV, KARANDE AD, HENG P. In-line quantification of drug and excipients in cohesive powder blends by near infrared spectroscopy [J]. Int J Pharm, 2010, 386(1-2):138
[8] HUBERT PH, NGUYEN-HUU JJ, BOULANGER B, et al. Harmonization of strategies for the validation of quantitative analytical procedures: a SFSTP proposal—part I[J]. J Pharm Biomed Anal, 2004, 36(3):579
[9] HUBERT PH, NGUYEN-HUU JJ, BOULANGER B, et al. Harmonization of strategies for the validation of quantitative analytical procedures A SFSTP proposal-Part II[J]. J Pharm Biomed Anal, 2007, 45(1):70
[10] HUBERT PH, NGUYEN-HUU JJ, BOULANGER B, et al. Harmonization of strategies for the validation of quantitative analytical procedures A SFSTP proposal-Part Ⅲ[J]. J Pharm Biomed Anal, 2007, 45(1):82
[11] WU ZS, XU B, DU M, et al. Validation of a NIR quantification method for the determination of chlorogenic acid in Lonicera japonica solution in ethanol precipitation process[J]. J Pharm Biomed Anal, 2012, 62(1):1
[12] MANTANUS J, ROZET E, BUTSELE KV, et al. Near infrared and Raman spectroscopy as Process Analytical Technology tools for the manufacturing of silicone-based drug reservoirs [J]. Anal Chim Acta, 2011, 699(1):96
[13] XUE Z, XU B, YANG C, et al. Method validation for the analysis of licorice acid in the blending process by near infrared diffuse reflectance spectroscopy[J]. Anal Methods, 2015, 7(14):5830
[14] ISO/DTS 21748 2017 Guidance for the use of repeatability, reproducibility and trueness estimates in measurement uncertainty evaluation [S]. 2017
[15] Eurachem Guide: Quantifying Uncertainty in Analytical Measurement[S]. 2000
[16] ISO. Guide to the Expression of Uncertainty in Measurement (GUM)[S]. 1995
[17] EA-4/16 EA Guidelines on the Expression of Uncertainty in Quantitative Testing[S]. 2004
[18] BARWICK VJ, ELLISON LR. VAM Project 3.2.1,development and harmonization of measurement uncertainty principles. Part D: Protocol uncertainty for evaluation from validation data[S]. 2000
[19] FEINBERG M, BOULANGER B, DEWE W, et al. New advances in method validation and measurement uncertainty aimed at improving the quality of chemical data[J]. Anal Bioanal Chem, 2004, 380(3):502
[20] SAFFAJ T, IHSSANE B, JHILAL F, et al. An overall uncertainty approach for the validation of analytical separation methods[J]. Analyst, 2013, 138:4677
[21] SAFFAJ T, IHSSANE B. Uncertainty profiles for the validation of analytical methods[J]. Talanta, 2011, 85(3):1535
[22] XUE Z, XU B, SHI XY, et al. Overall uncertainty measurement for near infrared analysis of cryptotanshinone in tanshinone extract[J]. Spectrochim Acta A Mol Biomol Spectrosc, 2017, 170: 39
[23] MEE R. β-expectation and β-content tolerance limits for balanced one-way ANOVA random model[J]. Technometrics, 1984, 26(3):251
[24] MEE R, OWEN DB. Improved factors for one-sided tolerance limits for balanced one-way ANOVA random model[J]. J Am Stat Assoc, 1983, 78(384):901
[25] TSUI KW, WEERAHANDI S. Generalized p-values in significance testing of hypotheses in the presence of nuisance parameters[J]. J Am Stat Assoc,1989, 84(406):602
[26] WEERAHANDI S. Generalized confidence intervals[J]. J Am Stat Assoc,1993, 88: 899
[27] HOFFMAN D, KRINGLE R. Two-sided tolerance intervals for balanced and unbalanced random effects models[J]. J Biopharm Stat, 2005, 15(2):283
[28] KENNARD RW, STONE LA. Computer aided design of experiments[J]. Technometrics, 1969,11:137
[29] Food and Drug Administration. Guidance for Industry, Bioanalytical Methods Validation[S]. 2001
[30] HOFFMAN D, KRINGLE R. A total error approach for the validation of quantitative analytical methods[J]. Pharm Res, 2007, 24(6):1157
[31] HOFFMAN D. Statistical considerations for assessment of bioanalytical incurred sample reproducibility[J]. AAPS, 2009, 11(3):570
[32] International Conference on Harmonization (ICH), Validation of analytical procedures: text and methodology, Q2 (R1). US, 1994-10-27 [EB/OL]. (2005-11) [2017-01-04]. http://www.ich. org/fileadmin/Public_Web_Site/ICH Products/Guidelines/Quality/Q2 R1/Step 4/Q2 R1 Guideline. pdf
