Isolation and Preparation of Myricitrin and Quercitrin from Hydrocotyle sibthorpioides Lam. by High-speed Counter-current Chromatography

High-speed counte-current chromatography(HSCCC)was developed for separation and purification of myricitrin and quercitrin from Hydrocotyle sibthorpioides Lam(HSL). The HSL ethanol extracts was extracted successively with petroleum ether,ethyl acetate and n-butanol. The ethyl acetate fraction was purified by Sephadex LH-20 elution with 75% methanol,and separated by high-speed countercurrent chromatography of n-hexane-n-butanol-water(1.75:1:1,v:v)two-phase solvent system,in which the upper phase as stationary phase and the lower phase was used as the mobile phase,at a flow rate of 2.0 mL/min.The rotation speed and detection wavelength were set at 900 r/min and 254 nm,respectively. The obtained product was subjected to high performance liquid phase detection and ¹H,¹³C NMR structural analysis. Results showed that,under this separation conditions,two products of 18.63 and 17.49 mg were obtained in one step,and the purity was 97.85% and 95.42%,respectivty,which were determined to be myricetin and quercetin by ¹H and ¹³C NMR.