Studies On The Key Enrichment Technoglogy Of Cordypin From Cordyceps Sinensis Fermentation Broth

Cordyceps militaris is a well-known traditional Chinese medicine which has the similar medicinal value as Cordyceps sinensis. Cordycepin is one of the major active compounds of C. militaris, which has a broad spectrum of biological activity, such as anti-virus and anti-tumor. Now, a large quantity of pure cordycepin is urgently needed for its further pharmacological studies and its medical products developments.Therefore, an efficient method for the enrichment and preparative separation of cordycepin from Cordyceps sinensis fermentation broth was developed.1, The cordycepin production of14different strains of C. militaris were investigated. Results indicated that the cordycepin production in fermentation broth of the C. militaris strain CM001was the highest by submerged culture. Furthermore, the effects of9different additives on cordycepin production by submerged culture of C.militaris CM001were investigated. The results showed that the production of cordycepin was increased by adenosine, adenine, glycine, guanine and alanine. Among them, adenine was the best addtive which resulted in the highest cordycepin production. One g/L adenine were added to the culture medium at the5th day of cultivation which resulted in a cordycepin production of220mg/L, exhibiting a10folds as high as that obtained in the basal medium culture.2,Macroporous adsorption resin was applied to the separation of cordycepin from C militaris fermentation broth. Adsorption and desorption characteristic of macroporous adsorption resins HPD-100,HPD-400,HPD-500,HPD-722,D101,AB-8,DM130for cordycepin in C. militaris fermentation broth were investigated. The results showed that HPD-100was the most appropriate resin for the separation of cordycepin in C. militaris fermentation broth. The conditions of separation and purification were primarily confirmed as follows:sample concentration was0.6mg/mL, sample loading amount was6BV, adsorption speed was3BV/h, desorption agent was25%ethanol, the volume of desorption agent was4BV and desorption speed was2BV/h. Sample was purified by HPD-100resin according to these technology parameters and the content of cordycepin could reach up to12.5%with8times as high as the fermentation broth dry powder.3,Application of high-speed counter-current chromatography (HSCCC) to the preparative purification of cordycepin from C.mililaris fermentation broth. The solvent systems for cordycepin separation were assessed and selected by partition coefficient and analytical high-speed counter-current chromatography.A high efficiency of HSCCC separation was achieved by a solvent system that consisted of ethyl acetate-n-butanol-0.5%ammonia water (2-3-5). Finally,43.8mg of cordycepin were obtained from400mg crude extracts in one-step separation with a purity of98.7%.