Extraction and purification of flavanone glycosides and kaemferol glycosides from defatted Camellia oleifera seeds by salting-out using hydrophilic isopropanol

Abstract

The purpose of this research was to investigate a salting-out procedure for isolating four flavonoid glycosides from defatted Camellia oleifera seeds. The procedure included extraction with 80% methanol, methanol removal and addition of an equal amount of hydrophilic isopropanol and salt to separate the isopropanol fraction from the water layer. Using successive column chromatography, kaemferol-3-O-[2-O-beta-D-glucopyransoyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 1), kaemferol-3-O-[2-O-beta-D-xylopyranosyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 2), naringenin-7-O-[beta-D-xylopyranosyl(1 -> 6)][beta-D-glucopyranosyl(1 -> 3)-alpha-L-rhamnopyranosyl(1 -> 2)]-beta-D-glucopyranoside (compound 3) and naringenin-7-O-beta-D-xylopyranosyl(1 -> 6)-beta-D-glucopyranoside (compound 4) were obtained. The structure of compound 3, a new flavanone glycoside, was analyzed using UV, FT-IR, (1)H NMR, (13)C NMR and HR-FAB-MS. Quantification using high-performance liquid chromatography (HPLC) demonstrated that defatted C. oleifera seed cake contains 7.92, 17.7, 2.23 and 1.06 mg/g of the four compounds. The extraction efficiencies of the four compounds were increased by 23.17%, 7.59%, 48.67% and 47.22% from those obtained with n-butanol partition extraction. This new extraction technique is simpler, faster and less expensive than the traditional extraction method. Accordingly, this salting-out method can potentially replace the existing one. (C) 2009 Elsevier B.V. All rights reserved.