Optimization of high-speed counter-current chromatography for separation of polyphenols from the extract of hawthorn (Crataegus laevigata) with response surface methodology

Separation and purification of polyphenols from the extract of hawthorn (Crataegus laevigata) was investigated with high-speed counter-current chromatography (HSCCC). Solvent systems were examined according to partition coefficients of target components and n-butanol-ethyl acetate-water (1:1:2, v/v/v) system was chosen. The critical experimental parameters of HSCCC were optimized and determined with response surface methodology (RSM) as following: Revolution speed was 850 rpm, flow-rate was 1.5 mL/min and temperature was 25 degrees C. Under these conditions, 500 mg of crude extract was introduced into the HSCCC system and a product of 9.7 mg with purity over 94.8% was obtained in 260 min, which was identified as chlorogenic acid with UV, LC-MS and H-1 NMR. The result proved that RSM is an efficient method to optimize HSCCC conditions, and HSCCC could be a very effective approach to separate and purify chlorogenic acid from natural source. (C) 2010 Elsevier B.V. All rights reserved.