高速逆流色谱法分离灵芝中的灵芝烯酸B

Separation of ganoderenic acid B from Ganoderma lingzhi by high-speed countercurrent chromatography

本实验建立一种快速从灵芝子实体中制备灵芝烯酸B的方法。以沪农灵芝1号子实体乙醇提取物为原料,经过D101大孔树脂粗分,用60%乙醇洗脱后,采用高速逆流色谱对获得的灵芝酸性三萜进行分离制备。确定最佳溶剂体系为正己烷-乙酸乙酯-甲醇-水(V/V/V/V,5:5:2:9),上相作为固定相,下相作为流动相,单因素法和正交实验设计确定高速逆流色谱法分离灵芝烯酸B的最佳工艺为:流速2.0mL/min,转速900r/min,一次上样量为500mg时,制备得到灵芝烯酸B化合物得率为(9.07±0.16)%,纯度为(85.04±3.45)%。用质谱、核磁对得到的灵芝烯酸B进行了结构鉴定。此法操作简单高效,为大量制备灵芝烯酸B提供了参考。

A rapid method for isolation of ganoderenic acid B from fruiting bodies of Ganoderma lingzhi is described.The extract of ganoderma triterpenoid acids(GTA)was obtained from G.lingzhi Hunong No.1 by using D101 macroporous resin as filler and the extract was eluted by 60%ethanol.The separation of obtained GTA sample was performed by high-speed countercurrent chromatography(HSCCC).The optimal solvent system was determined to be hexane-ethyl acetate-methanol-water(V/V/V/V,5:5:2:9)with the upper-layer as stationary phase and the sub-layer as mobile phase.The single-factor experiments and orthogonal experiments were carried out to determine the optimal processing for the preparation of ganoderenic acid B.Flow rate of 2.0 mL/min,rotational speed of 900 r/min,and loading quantity of 500 mg were proved to be optimal for the preparation process,and the yield of ganoderenic acid B reached(9.07±0.16)%and the purity reached(85.04±3.45)%.The structure of ganoderenic acid B was identified by nuclear magnetic resonance(NMR)and mass spectroscopy.This method is simple and efficient,and suitable to large scale preparation of ganoderenic acid B.