黑果小檗总花色苷的提取工艺及其急毒实验研究

Study on extraction process and acute toxicity of total anthocyanins from Berberis heteropoda Schrenk

目的建立黑果小檗总花色苷的最佳提取工艺,考察黑果小檗总花色苷的急性毒性。方法采用酶-超声波辅助法,以提取率为指标考察酶解pH值、酶解温度、酶解时间、酶用量对黑果小檗总花色苷提取的影响。将60只昆明种小鼠随机分为空白组、16.000g/kg、12.800g/kg、10.240g/kg、8.192g/kg和6.554g/kg剂量组,每组10只,采用一次性灌胃并观察小鼠14d内急性毒性反应,以死亡率为指标计算半数致死量(LD50)。结果黑果小檗总花色苷的最佳提取工艺:果胶酶酶解液pH值为4,酶解温度50℃,酶解时间30min,酶用量1.5%,在此条件下黑果小檗总花色苷的提取率最高,为1.45%。黑果小檗总花色苷提取物对小鼠的急性经口LD50为10.929g/kg,其95%置信区间为9.679~12.431g/kg。结论本方法简便、可靠、高效,适用于黑果小檗总花色苷的提取,急毒实验表明,黑果小檗总花色苷具有一定毒性,为临床安全用药提供实验参考。

Objective To investigate the optimal extraction process of total anthocyanins from the Berberis heteropoda Schrenk and to test the acute toxicity. Methods The extraction rate was used as an indicator to investigate the effects of enzymatic hydrolysis pH, enzymatic hydrolysis temperature, enzymatic hydrolysis time and enzyme dosage on the extraction of total anthocyanins from the Berberis heteropoda Schrenk by enzyme-ultrasonic-assisted method. 60 Kunming mice were randomly divided into blank group, 16.000 g/kg, 12.800 g/kg, 10.240 g/kg, 8.192 g/kg and 6.554 g/kg, with 10 rats in each group. The acute toxic reaction was observed within 14 days after the Single intragastric administration, and the half lethal dose （LD 50 ） was calculated based on the mortality rate. Results The optimum extraction conditions were as follows： enzymolysis pH of 4, enzymolysis temperature 50℃, enzymolysis time 30 min, enzyme dosage 1.5%. Under the condition, the extraction rate of total anthocyanins was the highest, at 1.45%. The results of acute toxicity test showed that the acute oral LD 50 of the total anthocyanins extract of Berberis heteropoda Schrenk was 10.929 g/kg, and the 95% confidence interval was 9.679-12.431 g/kg. Conclusion／ The method is simple, reliable and efficient, and is suitable for the extraction of total anthocyanins from Berberis berghei. Acute toxicity text shows that the total anthocyanins of Berberis heteropoda Schrenk has certain toxicity, which could provide an experimental reference for clinical safety.