Standard Deliberation

Uncertainty profile for validating the HPLC quantitative analysis method of rhubarb

Expand
  • 1. National Institutes for Food and Drug Control, Beijing 100050, China;
    2. School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou 510006, China;
    3. Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State Administration of Traditional Chinese Medicine, Guangzhou 510006, China;
    4. Guangdong Academies Traditional Chinese Medicine Quality Engineering Technology Research Center, Guangzhou 510006, China

Received date: 2021-10-26

  Online published: 2024-06-25

Abstract

Objective: To develop a validation method based on the uncertainty profile (UP) and β-content tolerance intervals (β-CTI), and apply the proposed method to validate the high performance liquid chromatography (HPLC) quantitative analysis for rhubarb. Methods: Taking the resolution of five chromatographic peaks of aloe-emodin, rhein, emodin, chrysophanol and physcion as the targets, the chromatographic parameters including the column temperature, and the mobile phase ratio were optimized by using the one-factor experimental design. Then the“3×5×4” full factorial design was used to obtain the validation data of the HPLC quantitative analysis. The experiment included 4 concentration levels, and each concentration level was performed in 5 replicates on 3 different days. The analysis of variance was performed on the validation data. The uncertainty of the HPLC quantitative analysis for aloe-emodin, rhein, emodin, chrysophanol and physcion at different concentrations was evaluated. Set±10% as the acceptable limits, the uncertainty profiles for five components in rhubarb were constructed to validate the HPLC quantitative analysis method. Results: With methanol and 0.1% phospho-ric acid solution (83∶17) as mobile phases, the resolution of the chromatographic peaks for the five components in rhubarb was greater than 1.5 at 25 ℃. The calibration curves of aloe-emodin, rhein, emodin, chrysophanol, and physcion displayed good linearity over the ranges of 0.013-50.65, 0.879-43.05, 1.016-50.8, 1.002-50.1, 1.023-51.15 μg·mL-1, respectively. The uncertainties of HPLC quantitative analysis for aloe-emodin, rhein, emodin, chrysophanol and physcion were within acceptable limits (±10%) over the ranges of 2.032-40.64,1.758-35.16, 2.032-40.26, 2.004-40.08 and 2.046-40.92 μg·mL-1, respectively. Conclusion: The UP method is easy to operate and can comprehensively and scientifically evaluate the accuracy of the analytical method. Application of the UP in the validation stage can control the risk of using the analytical method in routine phase.

Cite this article

DAI Sheng-yun, ZHAN Shu-yi, OU-YANG Xiao-jie, MA Shuang-cheng, ZHENG Jian, SUN Fei . Uncertainty profile for validating the HPLC quantitative analysis method of rhubarb[J]. Chinese Journal of Pharmaceutical Analysis, 2021 , 41(12) : 2202 -2210 . DOI: 10.16155/j.0254-1793.2021.12.19

References

[1] The International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). Harmonized Tripartite Guideline, Q2 (R1), Validation of Analytical Procedures: Text and Methodology, Current Step 4 Version[EB/OL].https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf
[2] HUBERT P, 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, 2004, 36(3): 579
[3] 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, 380:502
[4] GONZALEZ AG, HERRADOR MA. Accuracy profiles from uncertainty measurements[J].Talanta, 2006, 70:896
[5] MARINI RD, CHIAP P, BOULANGER B, et al. LC method for the determination of R-timolol in S-timolol maleate: validation of its ability to quantify and uncertainty assessment[J].Talanta, 2006, 68:1166
[6] FEINBERG M. Validation of analytical methods based on accuracy profiles[J].J Chromatogr A, 2007, 1158:174
[7] DAI SY, XU B, ZHANG Y, et al. Establishment and reliability evaluation of the design space for HPLC analysis of six alkaloids in Coptis chinensis (Huanglian) using Bayesian approach[J].Chin J Nat Med, 2016, 9(14): 697
[8] DAY SY, XU B, ZHANG Y, et al. Robust design space development for HPLC analysis of five chemical components in Panax notoginseng saponins[J].J Liq Chromatogr RT, 2016, 39(10): 504
[9] GONZALEZ AG, HERRADOR MA. A practical guide to analytical method validation, including measurement uncertainty and accuracy profiles[J].Trac-Trend Anal Chem, 2007, 26(3): 227
[10] SAFFAJ T, IHSSANE B. Uncertainty profiles for the validation of analytical methods[J].Talanta, 2011, 85(3): 1535
[11] HASNAA H, TAOUFIQ S, AIMIN T, et al. Full validation using β-content, γ-confidence tolerance interval: application for LC-MS/MS determination of doxycycline in human plasma[J].Chemometr Intell Lab, 2017, 168: 89
[12] BOUCHAFRA H, EI KARBANE M, AZOUGAGH M, et al. Application of a new strategy of validation based on "β,γ-content tolerance interval" for checking the chiral chromatography method for quantification of the chiral impurity of levofloxacin[J].J Brazil Chem Soc, 2015, 27(3): 500
[13] XUE Z, XU B, SHI X Y, et al. Overall uncertainty measurement for near infrared analysis of cryptotanshinone in tanshinone extract[J].Spectrochim Acta A, 2017, 170: 39
[14] RUDAZ S, FEINBERG M. From method validation to result assessment: established facts and pending questions[J].Trac-Trend Anal Chem, 2018, 105: 68
[15] 刘佳, 李莉.不同道地产区大黄功效组分含量的差异分析[J].中国现代中药, 2017, 19(5): 662
LIU J,LI L. Analysis on the difference of effective components of Rhubarb in different regions[J].Mod Chin Med,2017, 19(5): 662
[16] 吴丽, 徐春蕾, 季易彦, 等. 大黄及大黄蒽醌类有效成分对胰腺腺泡细胞损伤的保护作用[J].中药新药与临床药理, 2014, 25(5): 590
WU L, XU CL, JI YY,et al. Protective effects of rhubarb and its anthraquinones on pancreatic acinar cell injury[J].Tradit Chin Drug Res Clin Pharmacol, 2014, 25(5): 590
Outlines

/