Box-Behnken设计-效应面法优化白花蛇舌草总黄酮的提取工艺

目的: 优选白花蛇舌草黄酮类成分的提取工艺,为该药材的资源开发提供参考。方法: 以乙醇体积分数、乙醇用量及提取时间为自变量,芦丁、异槲皮苷、槲皮苷提取量的总评"归一值"为因变量,通过Design-Expert 8.0软件对自变量各水平进行多元线性回归和二项式拟合,利用效应面法优选提取工艺并进行预测分析。采用 HPLC-DAD测定芦丁、异槲皮苷、槲皮苷含量,色谱条件为流动相乙腈(A)-0.05%磷酸溶液(B)梯度洗脱,检测波长254 nm。结果: 最佳提取工艺为加14倍量73%乙醇回流提取2次,每次1.5 h;芦丁、异槲皮苷、槲皮苷平均提取量分别为2.529,0.435,0.215 mg·g^-1,总评"归一值"实测值0.922 7与预测值偏差-1.24%。结论: 优化的提取工艺稳定可行,预测准确度高,为白花蛇舌草总黄酮的制剂开发提供参考。

Optimization of Extraction Process of Total Flavonoids from Oldenlandia diffusa by Box-Behnken Design-response Surface Methodology

Objective: To provide a reference for resource development of Oldenlandia diffusa by optimize extraction process of total flavonoids from this medicinal herb. Method: Extraction time, ethanol concentration and amount were selected as independent variables, overall desirability of contents of rutin, isoquercitrin and quercitrin as dependent variable, Box-Behnken design-response surface methodology was used to optimize extraction technology.HPLC-DAD was adopted to determine contents of rutin, isoquercitrin and quercitrin with mobile phase of acetonitrile (A)-0.05% phosphoric acid solution (B) for gradient elution and detection wavelength of 254 nm. Result: Optimal extraction process was as following:extracted 2 times with 14-fold the amount of 73% ethanol for 1.5 h per time.Average extracting amounts of rutin, isoquercitrin and quercitrin were 2.529, 0.435, 0.215 mg·g^-1, respectively.Deviation between the measured and predicted value of overall desirability was -1.24%. Conclusion: This extraction process is stable and feasible with high prediction accuracy, which can provide a reference for preparations development of total flavonoids from O. diffusa.