柴胡地上部分总黄酮的大孔吸附树脂分离纯化工艺及抑菌活性研究

Separation, purification and antibacterial activity of total flavonoids from the aboveground parts of Bupleurum chinense DC. using a macroporous adsorption resin

采用静态吸附-洗脱试验,考察了12种大孔吸附树脂对柴胡地上部分总黄酮的吸附和洗脱效果,从中筛选出最佳的树脂为NKA-II型大孔吸附树脂。利用动态吸附-洗脱试验,对NKA-II型吸附树脂分离纯化柴胡地上部分总黄酮的工艺条件进行了优化,得到最优的工艺参数为:树脂径高比为2∶7,上样质量浓度为0.08 g/mL(每毫升药液相当于生药0.08 g),上样流速为3.0 BV/h,上样量为16 BV,洗脱剂为70%乙醇,洗脱流速为2.5 BV/h,洗脱剂用量为10 BV。工艺放大验证结果表明,在最佳的分离纯化工艺条件下,吸附量、解吸率及干浸膏中总黄酮含量分别为80.46 mg/g、95.72%和65.71%,分离纯化的效果较好。测试了柴胡地上部分总黄酮经纯化后的抑菌活性,结果显示:总黄酮对2种细菌——表皮葡萄球菌(Staphylococcus epidermidis)和鼠伤寒沙门菌(Salmonella typhimurium)的抑菌活性较强,最低抑菌浓度(MIC)均为6.15 mg/mL;对2种真菌——球形马拉色菌(Malassezia globosa)和糠秕马拉色菌(Malassezia furfur)的抑菌活性较强,MIC分别为12.30 mg/mL和24.60 mg/mL。

Using static adsorption-elution tests, the adsorption and elution effects of 12 macroporous adsorption resins on total flavonoids from the aboveground parts of Bupleurum chinense have been investigated. NKA-II macroporous adsorption resin was found to have the best overall performance. Using dynamic adsorption-elution tests, the process conditions for the separation and purification of total flavonoids from the aboveground parts of Bupleurum chinense using NKA-II adsorption resin were optimized. The optimal process parameters were as follows: the ratio of diameter to height of the resin was 2∶7, the mass concentration of the sample was 0.08 g/mL(each milliliter of liquid is equivalent to 0.08 g of crude drug), the loading flow rate was 3.0 BV/h, the loading volume was 16 BV, the eluent was 70% ethanol, the elution flow rate was 2.5 BV/h, and the eluent dosage was 10 BV. The results of process scale-up showed that under the optimal separation and purification process conditions, the adsorption capacity, desorption rate and total flavonoid content in the dry extract were 80.46 mg/g, 95.72% and 65.71%, respectively, and the separation and purification effects were good. The antibacterial activity of the purified total flavonoids from the aboveground part of Bupleurum chinense was tested. The results showed that the total flavonoids had strong antibacterial activity against two bacteria, Staphylococcus epidermidis and Salmonella typhimurium, with a minimum inhibitory concentration(MIC) of 6.15 mg/mL in each case. The total flavonoids exhibited strong antifungal activity against two fungi, Malassezia globosa and Malassezia furfur, with MICs of 12.30 mg/mL and 24.60 mg/mL, respectively.