红豆杉枝叶中总生物碱提取工艺及其抗肿瘤活性研究

本论文以红豆杉枝叶为原料,通过单因素试验和响应面法优化了红豆杉枝叶中总生物碱的酶与超声波辅助耦合提取工艺,并对粗总生物碱进行了体外抗肿瘤活性实验。研究结果表明:酶与超声波辅助耦合提取总生物碱的最佳工艺条件为原料量3.00 g,pH值为3的19%乙醇溶液作溶剂、酶质量浓度0.10 g/L、液料比12∶1（mL∶g）、酶解时间52 min、酶解温度50℃和超声波功率150 W,在此条件下提取1次的总生物碱得率为0.126%。响应面模型显著性检验发现模型的P<0.000 1,表明差异极显著,回归方程有效,模型具有统计学意义。粗总生物碱萃取物的体外抗肿瘤活性结果表明:红豆杉枝叶粗总生物碱萃取物对3种肿瘤细胞都有较好的抑制作用,特别是对人宫颈癌（Hela）细胞的抑制活性最强。

This paper takes the branches and leaves of Taxus chinensis as raw materials, optimizes the enzyme- and ultrasound-assisted coupled extraction process of total alkaloids from Taxus chinensis branches and leaves through single-factor experiments and response surface methodology, and performs in vitro antitumor activity experiments on the crude total alkaloids. The research findings indicate that the optimal process conditions for enzyme- and ultrasound-assisted coupled extraction of total alkaloids are as follows: raw material dosage of 3.00 g, 19% ethanol solution with pH 3 as the extraction solvent, enzyme mass concentration of 0.10 g/L, solid-liquid ratio of 12:1 (mL:g), enzymatic hydrolysis time of 52 min, enzymatic hydrolysis temperature of 50℃, and ultrasonic power of 150 W. Under these conditions, the extraction yield of total alkaloids from a single extraction reaches 0.126%. The significance test of the response surface model reveals that the P-value of the model is less than 0.0001, demonstrating an extremely significant difference, the regression equation is valid, and the model has statistical significance. The in vitro antitumor activity results of the crude total alkaloid extract show that the crude total alkaloid extract from Taxus chinensis branches and leaves exhibits good inhibitory effects on three types of tumor cells, with the strongest inhibitory activity against human cervical cancer (Hela) cells.