The intake of isoflavones and the resulting equol contents of both plasma and milk of the same red clover-fed cows are reported for the first time in cyclic change-over design study. Cows were fed four different red clover silages and two timothy -meadow fescue silages as controls. The red clover silages contained daidzein, formononetin, biochanin A and genistein, whereas the timothy -meadow fescue silages contained no isoflavones. We found a strong association (y ¼ 0·071x þ 2·75, R 2 0·71) between the formononetin intake (x) and equol concentration (y) in the plasma, while the formononetin intake and milk equol concentration were weakly associated (y ¼ 0·0035x þ 0·358, R 2 0·20). This means that a small part of the total formononetin in the silage is secreted into milk as equol. The mean equol contents in plasma and milk of cows fed red clover silage diets were in the range of 4·6 -8·4 mg/l and 458 -643 mg/l, respectively, while the respective values for the control diets were in the range of 0·8 -1·5 mg/l and 171 -287 mg/l. We showed that shorter growing periods of red clover resulted in higher silage formononetin contents and plasma and milk equol contents, suggesting that the equol content of milk can be manipulated by varying the harvesting strategy of red clover. We conclude that milk equol is derived from the formononetin of red clover silage and that milk from red clover-fed cows can be considered as a source of equol in human nutrition.
The isoflavonoids, equol, formononetin, daidzein, genistein, biochanin A, and O-demethylangolensin (O-DMA), were analyzed from commercial cartons of skimmed Finnish milk by HPLC-diode array detector (DAD)-FL. We found 411 +/- 65 ng/mL of equol and traces of formononetin and daidzein in organic skimmed milk whereas conventionally produced milk contained 62 +/- 16 ng/mL of equol and no formononetin or daidzein.
This study is a summary of Finnish investigations of the phytoestrogen content of legume plants, red clover, white clover, alfalfa, and goat's rue. In addition to the chemical analyses, biological studies were performed. Uterine weight of immature rats was used as an indicator of the estrogenic effect of the fodder used. All red clover varieties studied contained estrogenic isoflavones, especially formononetin and biochanin-A. The phytoestrogen content varied from 1.0% to 2.5% of dry matter. The biological study of white clover showed a clear estrogenic effect not visible through chemical analysis. Alfalfa contains small quantities of formononetin and biochanin-A, but 25-65 ppm coumestrol in dry matter. The estrogenic effect of alfalfa was obvious in the biological study. Goat's rue did not contain any known phytoestrogens, and the biological study was completely negative.
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