Rapid separation and purification of two C25 steroids with bicyclic [4.4.1] A/B rings from the marine fungus Aspergillus sp. LS116 by high-speed counter-current chromatography in stepwise elution mode.

Two C25 steroids with bicyclic [4.4.1] A/B rings were successfully separated from the marine fungus Aspergillus sp. LS116 by a two-step high-speed counter current chromatography (HSCCC). Petroleum ether/ethyl acetate/methanol/water (5.5:11:5:7, v/v) and petroleum ether/ethyl acetate/methanol/water (5:6:5:7, v/v) were selected as two optimum two-phase systems to purify two C25 steroids, neocyclocitrinol B (1) and threo-23-O-methylneocyclocitrinol (2). The purity of two compounds was over 94%. Their structures were determined by comprehensive spectroscopic techniques. This is the first report about rapid separation and identification of C25 steroids with bicyclic [4.4.1] A/B rings by HSCCC.