响应面法优化黄金茶总黄酮提取工艺及对胰蛋白酶活性影响的研究

Optimization of Extraction Process of Total Flavonoids from Chimonanthus salicifolius S. Y. Hu. Leaves by Response Surface Methodology and Its Effect on Trypsin Activity

摘要:

以黄金茶为原材料，采用超声辅助法提取黄金茶的总黄酮，研究了超声时间、超声功率、乙醇体积分数、料液比对得率的影响。在此基础上，采用Box-Benhnken中心组合法进行4因素3水平试验设计，以黄金茶总黄酮得率为响应值，进行响应面分析，并初步探究经聚酰胺树脂纯化的黄酮对胰蛋白酶活性的影响。得到最优工艺条件为：超声时间30 min、超声功率57.59 W、乙醇体积分数52.03%、料液比1:23.85。在此工艺下总黄酮的得率为19.38%±0.12%，实际值与预测值之间的相对误差仅为1.9%，变异系数仅为0.6%，重复性比较好。同时建立了乙醇溶液提取黄金茶总黄酮的二次数学模型，对目标产物提取具有良好的预测作用。在此工艺条件下得到的总黄酮经聚酰胺树脂纯化，30%的乙醇洗脱浓缩。纯化后的黄酮对胰蛋白酶的半数抑制浓度（IC50）为1.36 mg/mL。

Abstract:

Using Chimonanthus salicifolius S. Y. Hu. leaves as raw material, the total flavonoids of Chimonanthus salicifolius S. Y. Hu. leaves were extracted by ultrasonic-assisted method. The effects of ultrasonic time, ultrasonic power, ethanol volume fraction, solid-liquid ratio on the yield were investigated. On the basis of single factor tests, Box-Benhnken center combination experiment was used to carry out four factors and three levels experiment design. The yield of total flavonoids of Chimonanthus salicifolius S. Y. Hu. leaves was used as the response value for the response surface analysis, and the effect of the flavonoids purified from polyamide resin on the trypsin activity was explored. The optimal process conditions were as follows: ultrasonic time 30 min, ultrasonic power 57.59 W, ethanol concentration 52.03% and solid-liquid ratio 1: 23.85. Under these conditions, the yield of total flavonoids was 19.38% ± 0.12%. The relative error between the actual value and the predicted value was only 1.9%, and the coefficient of variation was only 0.6%. The extraction method also showed good repeatability. A second mathematical model of total flavonoids extracted from ethanol solution was established, which has a good predictive value for the extraction of the target product. The total flavonoids obtained were purified by polyamide resin and concentrated by elution with 30% ethanol. The half-inhibitory concentration value (IC50 value) of the purified flavonoids on trypsin was 1.36 mg/mL.