School of Chemistry and Chemical Engineering, Central South University, Changsha, China.
J Sep Sci. 2013 Oct;36(20):3338-44. doi: 10.1002/jssc.201300653. Epub 2013 Sep 1.
An off-line 2D high-speed counter-current chromatography technique in preparative scale has been successfully applied to separate and purify the main compounds from the ethyl acetate extract of Desmodium styracifolium. A two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water at an optimized volume ratio of 1:2:1:2 v/v/v/v was used. Conventional high-speed counter-current chromatography was used as the first dimension, and the upper phase of the solvent system was used as the stationary phase in the head-to-tail elution mode at a flow rate of 2.0 mL/min and a rotation speed of 900 rpm. Recycling high-speed counter-current chromatography served as the second dimension to separate an impure fraction of the first dimension. A total of four well-separated substances including vanillic acid (1), β-sitosterol (2), formononetin (3), and aromadendrin (4) were obtained, and their purities and structures were identified by HPLC-MS and (1) H NMR spectroscopy. The results illustrated that off-line 2D high-speed counter-current chromatography is an effective way to isolate compounds in complex samples.
离线二维高速逆流色谱技术已成功应用于从 Desmodium styracifolium 的乙酸乙酯提取物中分离和纯化主要化合物。使用由正己烷/乙酸乙酯/甲醇/水组成的两相溶剂系统，优化的体积比为 1:2:1:2 v/v/v/v。采用常规高速逆流色谱作为第一维，在流速为 2.0 mL/min 和转速为 900 rpm 的情况下，以头到尾洗脱模式使用溶剂系统的上相作为固定相。回收高速逆流色谱作为二维分离第一维的不纯部分。总共分离得到四种分离良好的物质，包括香草酸（1）、β-谷甾醇（2）、芒柄花素（3）和阿马里新（4），并通过 HPLC-MS 和 1 H NMR 光谱鉴定了它们的纯度和结构。结果表明，离线二维高速逆流色谱是一种从复杂样品中分离化合物的有效方法。
