Isolation and purification of isoflavones from soy flour by high-speed countercurrent chromatography

Degenhardt, A.; Winterhalter, P.

European Food Research and Technology 213(4/5): 277-280

2001


ISSN/ISBN: 1438-2377
DOI: 10.1007/s002170100386
Accession: 003485360

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Abstract
High-speed countercurrent chromatography (HSCCC) was applied to the isolation of isoflavones from soy on a preparative scale. Two different protocols were developed for the isolation of monoglucosylated/acetylated isoflavones as well as malonylated derivatives. Analysis of malonylated derivatives requires a quick and gentle extraction/separation procedure and the use of acid-free solvents for HSCCC. An appropriate solvent system for the isolation of 6"-O-malonylgenistin is the biphasic mixture of tert-butyl-methyl ether/n-butanol/acetonitrile/water (1/3/1/5). In the case of monoglucosylated/acetylated isoflavones, an enrichment and clean-up step (i.e., adsorption on XAD-7 resin) is preferably carried out prior to the HSCCC separation. The separation is then achieved by the application of tert-butyl-methyl ether/acetonitrile/water (2/2/3) and tert-butyl-methyl ether/acetonitrile/water (6/3/8) as solvent systems. Peak purity of the isolated compounds was checked by HPLC-MS and 1H-NMR spectroscopy. The isolated compounds can be used for studies on metabolism and bioavailability of isoflavone derivatives.

Isolation and purification of isoflavones from soy flour by high-speed countercurrent chromatography