Application of high-speed counter-current chromatography coupled with high performance liquid chromatography for the separation and purification of Quercetin-3- O -sambubioside from the leaves of Nelumbo nucifera

Abstract

Quercetin-3-O-sambubioside [Quercetin-3-O-β-D-xylopyranosyl (1→2)-β-D-glucopyranoside] was separated and purified by semi-preparative high-speed counter-current chromatography (HSCCC) with a two-phase-solvent system composed of ethyl acetate-n-butanol–water (4∶1∶5, v/v) from the leaves of Nelumbo nucifera (Lotus). A total of 5.0mg of the targeted compound with a purity of 98.6% as determined by high performance liquid chromatography (HPLC) was obtained from 100m g of the crude extract cleaned up by AB-8 macroporous resin in a one-step separation. Quercetin-3-O-sambubioside was a novel flavonoid glycoside from the leaves of Nelumbo nucifera, and its chemical structure was identified by means of ESI-MS, 1D NMR and 2D NMR.