华中枸骨叶总黄酮的纯化及其抑菌活性研究

以吸附率、解吸率、回收率为考察指标,在单因素实验基础上,采用正交设计优化D101大孔树脂纯化华中枸骨叶总黄酮的工艺条件。同时采用牛津杯法,考察纯化前后的提取物对常见细菌的体外抑菌作用。D101大孔吸附树脂纯化华中枸骨叶总黄酮的最佳工艺条件为:总黄酮质量浓度为0.735 mg/mL,pH为1.93,3 BV上样液,径高比1:8.5,上样速度6 BV/h,上样完毕后,静置30 min;洗脱时,先用3 BV去离子水,再用70%乙醇8 BV,洗脱流速6 BV/h,得到总黄酮回收率为91.06%,总黄酮质量分数从48.52%增长到78.26%。体外抑菌结果显示:华中枸骨叶总黄酮纯化物对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌、宋内志贺菌的抑菌圈分别为14.07、17.15、7.12、7.57 mm。除大肠埃希菌外,对于其他3种菌的抑菌效果均是粗提物好于纯化物。

Purification and Antibacterial Activity of Total Flavonoids from the Leaves of Ilex centrochinensis S. Y. Hu

Used the adsorption rate,desorption rate and recovery rate as indexes,on the basis of single factor experiment,orthogonal design was used to optimize the technological conditions of purifying total flavonoids from leaves of Ilex centrochinensis S. Y. Hu by D101 macroporous resin. The antibacterial activity in vitro of total crude and purified flavonoids was studied by oxford cup test. The optimum technological conditions for purifying total flavonoids from leaves of Ilex centrochinensis S. Y. Hu by D101 macroporous resin are as follows:Total flavonoids concentration was 0.735 mg/mL,pH was 1.93,sample volume was 3 BV,diameter-height ratio was 1:8.5,adsorption velocity was 6 BV/h. It was stand still for 30 minutes before desorption,then 3 BV deionized water was used to remove impurities. Desorption ethanol(8 BV)concentration was 70%,desorption velocity was 6 BV/h. The recovery rate of total flavonoids was 91.06%,the mass fraction of total flavonoids was increased from 48.52% to 78.26%. The results of bacteriostasis in vitro showed that:The antimicrobial circles of total flavonoids from leaves of Ilex centrochinensis S. Y. Hu against Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Shigella sonnei were 14.07,17.15,7.12 and 7.57 mm,respectively. Except Escherichia coli,the antibacterial effect against the other 3 kind of bacteria of crude extract was better than the purified producton.