Abstract
A method of using high-speed counter-current chromatography (HSCCC) was established for preparative isolation and purification of epothilones from strain S. cellulosum Fim0810 fermentation broth. Epothilone A and B components were successfully purified for the first time by HSCCC with a two-phase solvent system composed of n-hexane-ethyl–acetate–methanol–water (7:7:5:5 by volume). In a single operation, 100 mg of the crude extract was separated to yield 22.8 mg of Epothilone A and 38.4 mg of Epothilone B with the purity of 97.2% and 96.5%, respectively. The HSCCC fractions were determined by high-performance liquid chromatography (HPLC), UV, ESI–MS, and 1H NMR analysis. This technique should also be applicable to the preparation of epothilone analogs from Sorangium cellulosum fermentation broth.
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