Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, and Department of Chemistry, Fuzhou University, Fuzhou, Fujian, PR China.
J Sep Sci. 2011 Feb;34(4):385-92. doi: 10.1002/jssc.201000577. Epub 2011 Jan 21.
Ergosterol and stigmasterol are the most common phytosterols in the traditional Chinese medicine. They are two major sterol compounds in Anoectochilus roxburghii (wall) Lindl (A. roxburghii) and have been proved to have many important biological activities. A method by using high-speed counter-current chromatography (HSCCC) has been successfully developed for separation and purification of ergosterol and stigmasterol in A. roxburghii simultaneously in this paper. The optimum conditions used in this method were as follows: The two-phase solvent system consisted of n-hexane-ethylacetate-butanol-methanol-water (3.5:0.3:0.5:2.5:0.3, v/v); the rotation speed was 900 rpm; the flow rate of the lower phase was 1.5 mL/min. About 36.5 mg of ergosterol and 43.6 mg of stigmasterol were obtained from 100 g of A. roxburghii. The purity of ergosterol and stigmasterol was examined to be 92.0 and 95.5%, respectively, by using HPLC. The chemical structures of these components were identified by UV spectra, FT-IR, MS, (1) H-NMR and (13) C-NMR. The results demonstrated that high-speed counter-current chromatography was a feasible method to separate and purify ergosterol and stigmasterol from the herb. This separation and purification method was more effective than many other conventional techniques.
麦角甾醇和豆甾醇是中药中最常见的植物甾醇。它们是铁皮石斛（Anoectochilus roxburghii (wall) Lindl）中两种主要的甾醇化合物，已被证明具有许多重要的生物活性。本文采用高速逆流色谱（HSCCC）成功地开发了一种同时分离和纯化铁皮石斛中麦角甾醇和豆甾醇的方法。该方法的最佳条件如下：两相溶剂系统由正己烷-乙酸乙酯-正丁醇-甲醇-水（3.5:0.3:0.5:2.5:0.3，v/v）组成；转速为 900 rpm；下相流速为 1.5 mL/min。从 100 g 铁皮石斛中可得到约 36.5 mg 的麦角甾醇和 43.6 mg 的豆甾醇。采用 HPLC 检测，麦角甾醇和豆甾醇的纯度分别为 92.0%和 95.5%。通过 UV 光谱、FT-IR、MS、(1)H-NMR 和 (13)C-NMR 鉴定了这些成分的化学结构。结果表明，高速逆流色谱是一种从该草药中分离和纯化麦角甾醇和豆甾醇的可行方法。这种分离和纯化方法比许多其他常规技术更有效。
