Preparative isolation and puriﬁcation of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high-speed counter-current chromatography

Mei Yang; Xin-Jun Xu; Chun-Yan Xie; Jie-Yun Huang; Zhi-Sheng Xie; De-Po Yang

doi:10.1016/j.jpha.2012.03.004

Preparative isolation and puriﬁcation of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high-speed counter-current chromatography

J Pharm Anal. 2012 Aug;2(4):258-263. doi: 10.1016/j.jpha.2012.03.004. Epub 2012 Mar 14.

Authors

Mei Yang^{1

2}, Xin-Jun Xu^{1

2}, Chun-Yan Xie^{1

2}, Jie-Yun Huang^{1

2}, Zhi-Sheng Xie^{1

2}, De-Po Yang^{1

2}

Affiliations

¹ School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou 510006, China.
² Guangdong Technology Research Center for Advanced Chinese Medicine, Guangzhou 510006, China.

Abstract

An efﬁcient method for the isolation and puriﬁcation of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high speed counter-current chromatography (HSCCC) was established in this paper. The ether extracts of Radix Eupatorii Chinensis were puriﬁed by HSCCC with a solvent system of hexyl hydride-ethyl acetate-methanol-water (1:2:1:2, v/v/v/v). The upper phase was used as the stationary phase and the lower phase as the mobile phase. About 8.4 mg of 12,13-dihydroxyeuparin was obtained from 200 mg of ether extracts from Radix Eupatorii Chinensis in one-step HSCCC separation, with the purity of 96.71%, as determined by HPLC. After methanol-water recrystallization, the purity of 12,13-dihydroxyeuparin reached 99.83%. Such a simple and effective method was fairly useful to prepare pure compound as reference substances for related study on Radix Eupatorii Chinensis.

Keywords: 12,13-dihydroxyeuparin; High speed counter-current chromatography (HSCCC); Radix Eupatorii Chinensis.