海星中具有生物活性总皂苷的分离纯化

Separation and purification of bioactive asterosaponins from starfish

目的利用稻瘟霉生物活性成分跟踪筛选模型,对海星中皂苷进行提取和分离纯化。方法用65%EtOH对粉碎的海星(干)进行回流提取,减压浓缩后均匀分散于水中,用石油醚和正丁醇进行萃取,浓缩正丁醇得海星总皂苷,对其进行稻瘟霉生物活性测试。海星总皂苷粗提物通过正相硅胶、反相硅胶及葡聚糖凝胶LH-20柱层析进行分离纯化。用薄层层析法(TLC)检测把相同的洗脱液收集在一起,得到所需要的有效部位。结果与结论通过稻瘟霉生物活性筛选知海星乙醇粗提物、海星总皂苷均具有抗有丝分裂活性,而石油醚粗提物没有显著的活性。海星总皂苷通过正相硅胶和反相硅胶柱层析分离得到D1～D66个含有海星皂苷的组分,其中D1、D3、D4、D5、D6在TLC上呈现出单一斑点。D2进一步通过葡聚糖凝胶LH-20柱层析进行分离,得到收率约为81.2%的组分a,收率约为3%的组份b。

Objective To extract and isolate asterosaponins with biological activity from starfish.Methods Extracted with 65% ethanol from the dry starfish cuted into pieces and followed by filtration.The extracts were evaporated at low pressure and scattered in water,then extracted with petroleum ether and n-butyl alcohol.The n-butyl alcohol extracts were evaporated to gain asterosaponins.Conidia of Pyricularia oryzae for the screening of bioactive constituents from starfish was used.The asterosaponins were subjected to different chromatographic methods including silica gel column,ODS RP-18.By TLC,collected the same eluent to abtain the different components.Results and Conclusion It was found that ethanol crude extracts and n-butyl alcohol extracts had antimitotic constituents,however petroleum ether extracts had no significant activity.Six components were achieved by silica gel column and ODS RP-18,named D1~D6.Among them,D1,D3,D4,D5,D6 present single spot by TLC.D2 was further separated by sephadex LH-20 gel chromatography,and gained a（81.2%） and b（3%）.