Separation of Steroidal Constituents of Oviductus Ranae by One-Step Method High-Speed Counter-Current Chromatography

Abstract

The dichloromethane extracts of Oviductus Ranae were treated simply and two steroidal compounds were prepared by means of high-speed counter-current chromatography (HSCCC) directly. The two-phase solvent system was composed of petroleum ether–ethyl acetate–butanol–methanol–water (3.5:0.3:0.5:2.5:0.4, v/v/v/v/v). The upper phase was used as stationary phase and the lower phase was used as mobile phase. The dichloromethane extracts of Oviductus Ranae were separated and purified by utilizing the solvent system above. High-performance liquid chromatography analysis showed that the purity of each of the two compounds had reached 95%. Their chemical structures were confirmed as 7-hydroxycholesterol and 7-ketocholesterol according to the ¹H-NMR and ¹³C-NMR data in literature. The high-speed counter-current chromatography proposed in this paper can be used to prepare the main steroidal components in Oviductus Ranae. The method also has advantages in simplicity, speed, and efficiency, which can be applied to the preparation of the high purity 7-hydroxycholesterol and 7-ketocholesterol. In this study, HSCCC was used to separate the substances in Oviductus Ranae for the first time, which provides a foundation for the separation of Oviductus Ranae by means of HSCCC.

Keywords:

• 7-hydroxycholesterol
• 7-ketocholesterol
• HSCCC
• Oviductus Ranae

Acknowledgments

Many thanks to Changchun University of Chinese Medicine and Jilin University for providing the research offices.