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Isolation and preparation of cordycepin from Cordyceps militaris fermentation broth by high speed counter-current chromatography
HU Xia1，2 XIE Hong-qi1，3 LUO Wei1，2 XIA Zhi-lan1，2 LIU Dong-bo1，2
1.College of Horticulture and Landscape，Hunan Agricultural University，Changsha 410128，China；
2.Sub-health Intervention Technology Laboratory，State Administration of Traditional Chinese Medicine，Changsha 410128，China；
3.Key Laboratory for Crop Germplasm Innovation and Utilization of Hunan Province，Changsha 410128，China.
Abstract：Objective To develop a new method for the isolation and preparation of cordycepin by high speed counter-current chromatography(HSCCC)with Cordyceps militaris fermentation broth as raw material.Methods The solvent systems for cordycepin separation were assessed and selected by HPLC partition coefficient method and analytical HSCCC.A solvent system that consisted of ethyl acetate- n-butanol-0.5%ammonia water(2∶3∶5)was applied to the separation.The upper phase was used as the stationary phase，while the lower phase was used as the mobile phase for the cordycepin separation by HSCCC.Results A high efficiency of HSCCC separation was achieved.Finally，cordycepin(43.8 mg)was obtained from 400 mg crude extract of C.militaris fermentation broth in one-step separation with a purity of 98.7%.Conclusion HSCCC is an efficient and simple method for the largescale preparation of cordycepin from C.militaris fermentation broth.
Keywords：cordycepin；high speed counter-current chromatography；Cordyceps militaris(Fr.)Link；partition coefficient；material basis