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目的:优选荆芥穗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:选择乙醇体积分数、提取时间及乙醇用量为自变量,橙皮苷、木犀草素、芹菜素提取量的总评“归一值”为因变量,通过星点试验优选提取工艺,运用DesignExpert 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测定橙皮苷、木犀草素、芹菜素含量,流动相乙腈(A)-0.4%磷酸溶液(B)梯度洗脱(0~20 min,5%~20%A;20~50 min,20%~30%;50~60 min,30%~5%A),检测波长283 nm。结果:最佳提取工艺为加15倍量73%乙醇回流提取2次,每次2 h;橙皮苷、木犀草素、芹菜素提取量分别为3.991,1.307,0.589 mg·g-1,总评“归一值”0.925 3,与预测值的偏差-2.14%。结论:优选的提取工艺简单易行、准确度高,适用于荆芥穗总黄酮的工业化生产。
Objective: To optimize the extraction process of flavonoids in Nepeta euphorbia, and to provide reference for the resource development of this herb. Methods: The volume fraction of ethanol, the extraction time and the amount of ethanol as independent variables, hesperidin, luteolin, apigenin total amount of extraction of “normalized value” as the dependent variable, through the star point optimization of the extraction process, the use of DesignExpert 8.0 software performs multivariate linear regression and binomial fitting on each level of the independent variables, and optimizes the extraction process and predictive analysis by using response surface methodology. The contents of hesperidin, luteolin and apigenin were determined by HPLC. The mobile phase consisted of gradient elution with acetonitrile (A) - 0.4% phosphoric acid solution (B) (0-20 min, 5-20% 20% ~ 30%; 50 ~ 60 min, 30% ~ 5% A), detection wavelength 283 nm. Results: The optimum extraction process was extraction with 15 times the amount of 73% ethanol twice for 2 h each time. The extraction rates of hesperidin, luteolin and apigenin were 3.991, 1.307 and 0.589 mg · g-1 respectively. “Normalized value” 0.925 3, -2.14% deviation from the predicted value. Conclusion: The optimal extraction process is simple, accurate and suitable for the industrial production of total flavonoids of Nepeta.