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

Abstract

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 850rpm, flow-rate was 1.5mL/min and temperature was 25°C. Under these conditions, 500mg of crude extract was introduced into the HSCCC system and a product of 9.7mg with purity over 94.8% was obtained in 260min, which was identified as chlorogenic acid with UV, LC–MS and 1H 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.