银杏外种皮银杏酸的大孔树脂及高速逆流色谱联合分离

Separation and purification for ginkgolic acid from Ginkgo biloba L. sarcotesta by macroporous adsorption resin and high-speed countercurrent chromatography

银杏外种皮中含有丰富的银杏酚酸类物质,提取后采用大孔树脂和高速逆流色谱联合分离制备高纯度银杏酸单体组分,可以明显提高其利用价值。试验表明,最佳纯化条件为:DM130大孔树脂为吸附树脂,60 mL银杏酸提取液动态吸附2 h,100 mL 75%乙醇溶液动态解吸2 h,银杏酸的纯度可提高到40.18%;以正己烷:乙酸乙酯:甲醇:水(5:1:5:1,v/v)为溶剂体系,上相为固定相、下相为流动相,流速2 mL/min,转速800 r/min,高速逆流色谱进一步分离得到3个色谱峰。液相色谱分析表明,3个色谱峰的主要成分分别为C13:0、C15:1和C17:1,面积百分比分别达到了81.5%、95.1%和98.3%,具有良好的单体组分开发利用前景。结果表明大孔树脂联合高速逆流色谱可有效地分离纯化银杏酸组分。

There were rich in ginkgo acids in Ginkgo biloba L.sarcotesta,and the high purity ginkgolic acid monomer components were prepared by high-speed countercurrent chromatography(HSCCC)on the basis of macroporous adsorption resin purification,which can obviously improve its utilization value.The optimum process conditions of macroporous resin to purify ginkgolic acid were studied through orthogonal experiment on the basis of single factor experiments.The results showed that the purity of gingko acid could be increased to 40.18%.And the optimal conditions were identified as the follows,DM130 macroporous resin as excellent adsorption resin of ginkgo acid;dynamic adsorption time for 2 hours with ginkgo acid extract 60 mL;dynamic desorption time for 2 hours with 75%ethanol as the desorption reagent 100 mL.Three chromatographic peaks were isolated by high-speed countercurrent chromatography for further purification,and the separation conditions were identified as the follows,n-hexane:ethyl acetate:methanol:water(5:1:5:1,v/v)as solvent system,the above phase as stationary phase,the lower phase as mobile phase,the flow rate as 2 mL/min,and the speed as 800 r/min.The results of HPLC analysis showed that the main components of the three peaks were C13:0,C15:1 and C17:1,respectively,with an area percentage of 81.5%,95.1%and 98.3%,showing a good prospect for the development and utilization of monomer components.The results showed that macroporous resin combined with high speed countercurrent chromatography could effectively separate and purify ginkgo acid.