Abstract: Preparative high speed counter-current chromatography (HSCCC) was applied for the isolation and purification of a diorcinol from the fungus Aspergillus versicolor DJ013 regulated by epigenetic manipulation for the first time using stepwise elution with the two-phase solvent system composed of petroleum ether-ether acetate-methanol-water at (4∶5∶4∶5,v/v).The preparative HSCCC separation was performed on crude sample yielding diorcinol in a one-step separation,with purities over 97% as determined by UPLC.The temperature was 25 ℃ and the revolution speed was 800 rpm with a flow rate of 5.0mL/min.The eluent was continuously monitored by a UV monitor at 220 nm.The structure of this compound was identified by ESI-MS and NMR.Compared with the 48-hour preparation period combined with column chromatography and HPLC,the high speed countercurrent chromatography only needs 55 min,and the efficiency is greatly improved.Our results demonstrated that HSCCC was an efficient technique to separate diorcinol,which provide an approach to solve the problem of their sample availability for drug development.

Key words: Aspergullus versicolor, diphenyl ether, high speed counter-current chromatography