PREPARATIVE SEPARATION OF HIGH-PURITY CORDYCEPIN FROM CORDYCEPS MILITARIS(L.) LINK BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY

A high-speed counter-current chromatography (HSCCC) technique in a preparative scale has been applied to separate and purify cordycepin from the extract of Cordyceps militaris(L.) Link by a one-step separation. A high efficiency of HSCCC separation was achieved on a two-phase solvent system of n-hexane-n-butanol-methanol-water (23:80:30:155, v/v/v/v) by eluting the lower mobile phase at a flow rate of 2mL/min under a revolution speed of 850rpm. HSCCC separation of 216.2mg crude sample (contained cordycepin at 44.7% purity after 732 cation-exchange resin clean-up) yielded 64.8mg cordycepin with purity of 98.9% and 91.7% recovery. Identification of the target compound was performed by UV, IR, MS, 1H NMR, and 13C NMR.