Dai Sheng-Yun, Xu Bing, Zhang Yi, Li Jian-Yu, Sun Fei, Shi Xin-Yuan, Qiao Yan-Jiang
Research Center of Traditional Chinese Medicine Information Engineering, Beijing University of Chinese Medicine, Beijing 100029, China.
Research Center of Traditional Chinese Medicine Information Engineering, Beijing University of Chinese Medicine, Beijing 100029, China; Engineering Research Center of Key Technologies for Chinese Medicine Production and New Drug Development, Ministry of Education, Beijing 100029, China.
Chin J Nat Med. 2016 Sep;14(9):697-708. doi: 10.1016/S1875-5364(16)30083-8.
Coptis chinensis (Huanglian) is a commonly used traditional Chinese medicine (TCM) herb and alkaloids are the most important chemical constituents in it. In the present study, an isocratic reverse phase high performance liquid chromatography (RP-HPLC) method allowing the separation of six alkaloids in Huanglian was for the first time developed under the quality by design (QbD) principles. First, five chromatographic parameters were identified to construct a Plackett-Burman experimental design. The critical resolution, analysis time, and peak width were responses modeled by multivariate linear regression. The results showed that the percentage of acetonitrile, concentration of sodium dodecyl sulfate, and concentration of potassium phosphate monobasic were statistically significant parameters (P < 0.05). Then, the Box-Behnken experimental design was applied to further evaluate the interactions between the three parameters on selected responses. Full quadratic models were built and used to establish the analytical design space. Moreover, the reliability of design space was estimated by the Bayesian posterior predictive distribution. The optimal separation was predicted at 40% acetonitrile, 1.7 g·mL(-1) of sodium dodecyl sulfate and 0.03 mol·mL(-1) of potassium phosphate monobasic. Finally, the accuracy profile methodology was used to validate the established HPLC method. The results demonstrated that the QbD concept could be efficiently used to develop a robust RP-HPLC analytical method for Huanglian.
黄连是一种常用的传统中药，生物碱是其中最重要的化学成分。在本研究中，首次基于质量源于设计（QbD）原则开发了一种等度反相高效液相色谱（RP-HPLC）方法，用于分离黄连中的六种生物碱。首先，确定了五个色谱参数以构建Plackett-Burman实验设计。通过多元线性回归对关键分辨率、分析时间和峰宽进行响应建模。结果表明，乙腈百分比、十二烷基硫酸钠浓度和磷酸二氢钾浓度是具有统计学意义的参数（P < 0.05）。然后，应用Box-Behnken实验设计进一步评估这三个参数对选定响应的相互作用。建立了完全二次模型并用于建立分析设计空间。此外，通过贝叶斯后验预测分布估计设计空间的可靠性。预测在40%乙腈、1.7 g·mL⁻¹十二烷基硫酸钠和0.03 mol·mL⁻¹磷酸二氢钾条件下可实现最佳分离。最后，采用准确度轮廓法对建立的HPLC方法进行验证。结果表明，QbD概念可有效地用于开发一种稳健的黄连RP-HPLC分析方法。
