高速逆流色谱分离纯化阿尔泰金莲花中的活性成分

摘要

目的 研究阿尔泰金莲花乙醇提取物的抗氧化活性,通过高速逆流色谱分离其活性成分.方法 采用回流提取法,料液比为1:20,70％乙醇100℃回流提取2次,每次1 h,过滤,合并滤液得到阿尔泰金莲花乙醇提取物,以Vc为阳性对照,测定阿尔泰金莲花乙醇提取物对DPPH自由基清除作用.采用高速逆流色谱法,以乙酸乙酯-正丁醇-水(2:1:3,V:V:V)为溶剂系统,上相为固定相,下相为流动相,分离纯化阿尔泰金莲花中的活性成分,并对其活性成分进行质谱分析.结果 阿尔泰金莲花乙醇提取物具有较好的DPPH清除能力,从中分离纯化得到3个化合物,分别为荭草素2″-O-b-L-半乳糖苷(Ⅰ)、槲皮素-3-O-b-D-吡喃木糖-(1→2)-b-D-吡喃葡萄糖苷(Ⅱ)、荭草素(Ⅲ).结论 阿尔泰金莲花乙醇提取物具有较好的抗氧化活性,化合物Ⅰ和Ⅱ均为首次从该植物中分离得到.%Objective To investigate the antioxidant effect of ethanol extraction from Trollius altaicus C.A.Mey; and to isolate the constituents by high-speed counter-current chromatography(HSCCC).Methods The extract from Trollius altaicus C.A.Mey.by refluxing extract method,the ethanol concentration was 70％,solid-liquid ratio was 1:20 and the extraction time was 1.5 h for 2 times in the 100℃.Antioxidant activities were measured by DPPH free radicals scavenging experiments,the results were compared to the Vc which was used as positive control group.Active constituents were separated by HSCCC.Solvent system of ethyl acetate: n-butanol: water(2:1:3)was employed,the upper phase was stationary phase,the upper phase as stationary phase,the lower phase as mobile phase.And the chemical composition was determined by MS.Results The extracts from Trollius altaicus C.A.Mey.showed good DPPH free radicals scavenging ability.Three compounds were separated from T.altaicus as follows: orientin-2″-O-β-L-galactopyranosylide(Ⅰ),quercetin-3-O-β-D-xylopyranosyl(1→2)-β-D-glucopyranoside(Ⅱ)and oientin(Ⅲ).Conclusion Trollius altaicus C.A.Mey.was shown to have good antioxidant activities,compoundsⅡand Ⅲ were for the first time separated from this plant.