Preparative Isolation of Antioxidant Flavonoids from Small Black Soybeans by Centrifugal Partition Chromatography and Sequential Solid-Phase Extraction

Title
Preparative Isolation of Antioxidant Flavonoids from Small Black Soybeans by Centrifugal Partition Chromatography and Sequential Solid-Phase Extraction
Authors
전제승강석우엄병헌김철영
Keywords
dual-mode CPC; Rhynchosia volubilis; epicatechin; isoflavonoids; solid-phase extraction
Issue Date
2014-11
Publisher
Separation science and technology
Citation
VOL 49, NO 17, 2756-2764
Abstract
Unlike other members of the leguminous family that are generally used as food, Rhynchosia volubilis, a small soybean with a black seed coat, is used as a folk medicine in Korea. Using an on-line high performance liquid chromatography-2,2ʹ-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation screening system, one peak in the ethyl acetate fraction exhibited outstanding antioxidant capacity. To purify and determine the antioxidant compounds associated with this peak, we developed a centrifugal partition chromatography (CPC) method. The ethyl acetate fraction (500 mg) was subjected to CPC using a two-phase solvent system of n-hexane–ethyl acetate–ethanol–water (0.1:8:2:7, v/v) with a dual-mode CPC (descending-ascending order) in a bid to separate glycitin (3.1 mg), daidzin (14.4 mg), 6ʹʹ-O-acetyl genistin (6.9 mg), 6ʹʹ-O-acetyl daidzin (5.3 mg), and a mixture of epicatechin and genistin. The mixture was further separated through solid-phase extraction to yield epicatechin (6.8 mg) and genistin (23.5 mg), respectively. Structural identification of the R. volubilis flavonoids was performed by NMR and ESI-MS.
URI
http://pubs.kist.re.kr/handle/201004/48774
ISSN
01496395
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KIST Publication > Article
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