Abstract: An efficient method was established for the separation and purification of naphthoquinones from Eleutherinemericana. The naphthols was extracted with 95% ethanol from E.mericana and then enriched by extracting with ethyl acetate.The crude naphthol extract were separated by High-speed Counter-current Chromatography (HSCCC) with petroleum ether-ethyl acetate-methanol-water (6:4:5:5,v/v) as the solvent system.The revolution speed was 850 rpm with a flow rate of 2.0 mL/min.The eluent was continuously monitored by a UV monitor at 254 nm.Finally,87 mg of eleutherin and 35 mg of isoeleutherin were obtained from 200 mg crude extract with purities of 97.3%and 98.6% by HPLC analysis,respectively.Their structures were identified by mass spectrometry (MS) and nuclear magnetic resonance spectrum (NMR).The result showed that this method was simple,efficient and suitable for the isolation and separation of naphthols from E.mericana.

Key words: High-speed Counter-current Chromatography, Eleutherine mericana, eleutherin, isoeleutherin