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Separation and purification of ergosterol and stigmasterol in Anoectochilus roxburghii (wall) Lindl by high‐speed counter‐current chromatography
Author(s) -
Huang Liying,
Cao Yangyuan,
Xu Hui,
Chen Guonan
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000577
Subject(s) - stigmasterol , ergosterol , chromatography , chemistry , countercurrent chromatography , sterol , high performance liquid chromatography , biochemistry , cholesterol
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.