百蕊草有效成分提取工艺优化及生物活性检测

Optimization of the extraction of effective constituent of Thesium chinense Turcz and biological activity detection

为进一步拓展天然药物的应用领域,以百蕊草为原料,采用单因素和响应曲面法优化百蕊草有效成分提取工艺条件,测定提取物抗氧化性、生物安全性、抑菌性、抗炎性。结果表明:在乙醇浓度为65%、料液比为1∶40 g/mL、温度为65℃、超声时间为50 min的条件下,有效成分提取当量(以黄酮提取当量计)为28.2 mg/g;经萃取及层析纯化后的提取物在400μg/mL时对DPPH、ABTS自由基的清除率分别为93.5±0.31%、98.96±0.33%;对大肠杆菌和金黄色葡萄球菌的最低抑菌浓度为10~12.5 mg/mL;在1 mg/mL浓度范围内无细胞毒性,对巨噬细胞具有促增殖作用,能够抑制LPS诱导RAW264.7分泌NO,表现出良好的抗炎活性。研究结果说明提取物在1 mg/mL浓度范围内仍具有较好的生物活性,能为百蕊草活性成分的开发应用提供参考。

In order to expand the application of natural medicine,the extraction process of the effective components from Thesium chinense Turcz is optimized by single factors and the response surface methodology.The inoxidizability,biosafety,bacteriostasis and anti-inflammatory activity of the extract are determined.Results show that,the extraction equivalent of active ingredients(calculated by flavonoid extraction equivalent) is 28.2 mg/g under the conditions of an ethanol concentration of 65%,a solid-liquid ratio of 1∶40 g/mL,a temperature of 65 ℃ and an ultrasonic time of 50 min.The scavenging rates of free radicals of DPPH and ABTS are 93.5±0.31% and 98.96±0.33% respectively when the extract purified by extraction and chromatography is 400 μg/mL.The MICs for Escherichia coli and Staphylococcus aureus are 10~12.5 mg/mL.It has no cytotoxicity in the concentration range of 1 mg/mL,and facilitates the proliferation of macrophages.It can inhibit the secretion of NO by LPS-induced RAW264.7,and exhibits good anti-inflammatory activity.The results show that the extract has a good bioactivity in the concentration range of 1 mg/mL,which provides theoretical reference for the development and application of the active ingredients of Thesium chinense Turcz.