Identification of isoflavonoids in beer

The isoflavonoids, genistein (4',5,7-trihydroxyisoflavone), biochanin A (5,7-dihydroxy-4'-methoxyisoflavone), daidzein (4',7-dihydroxyisoflavone), and formononetin (7-hydroxy-4'-methoxyisoflavone) are supposed to be health-promoting dietary factors of plant origin. They are particularly abundant in seeds and other parts of many plant species belonging to Leguminosae. The most popular source of isoflavonoids in human diet is soy. Here, evidence is presented that isoflavonoids are regularly found in beer. Diethyl ether extracts of beer were fractionated on thin-layer chromatography-silica, (straight phase) and rechromatographed using a reversed phase high-performance liquid chromatography octadecylsilica column. The fractions were analyzed by two recently developed radioimmunoassays, the first of them being specific for diadzein/formononetin and the second one specific for genistein/biochanin A. The immunoreactivity was found only in fractions with the mobility corresponding to the positions of standards on control chromatograms. Additionally, 26 samples of bottled beer were analyzed for isoflavonoid content using the combination of reversed phase high-performance liquid chromatography and radioimmunoassay. The sum of the four isoflavonoids ranged from 1.26 to 29 nmol/L in individual beers. Formononetin was the major isoflavonoid (0.19-14.99 nmol/L), whereas the concentration of daidzein was several times lower (0.08-2.5 nmol/L). Genistein and biochanin A concentrations were comparable, ranging from 0.169-6.74 nmol/L and from 0.820-4.84 nmol/L for genistein and biochanin A, respectively. It is concluded that beer contains significant amounts of biologically active isoflavonoid phytoestrogens.