Forage Estrogens, Detection of Daizein, Formononetin, Genistein, and Biochanin A in Forages

Abstract

Daidzein, not previously reported in forages, was found in alfalfa, and ladino, red, and subterranean clovers. Formononetin and genistein, isolated earlier from red and subterranean clovers, have now also been found in alfalfa and ladino clover. Biochanin A, previously reported in red and subterranean clovers, was found in alfalfa but not in ladino clover. A combination of three sequential purifications on paper chromatograms followed by one on silicic acid chromatostrips was used to isolate the isoflavones. The identity of the compounds was confirmed by absorption spectra as well as by comparison of paper and silicic acid chromatograms of the isolated compounds with those of authentic samples of the isoflavones.NTEREST in naturally occurring forage

Significance of phenolic compounds in plant‐soil‐microbial systems

“…The isoflavonoids daidzein, formononetin, genistein, and biochanin A can be detected in forages using paper chromatography employing ethanol-chloroform (1:1), acetone-water (3:7), butanol-acetic acid-water (14:1:5), and many other solvent systems. 132 High pressure liquid chromatography (HPLC) isolation and gas chromatography mass spectrometry (GC-MS) identification of genistein and daidzein from soybean roots was reported 133 using methanolwater gradients. Separation of ferulic acid esters and phenolic acids on HPLC using a C18 column was achieved by using the acetonitrile-water (60:40) solvent system.…”

Isolation Techniques for Flavonoids

“…These flavonoids are generally chromatographed using less polar solvents than those required for the common flavones and flavonols. For example, the isoflavones daidzein, formononetin, genistein and biochanin A have been separated using chloroform-methanol (92 : 8, 3 : 1 and 1 : 1) (Guggolz et al, 1961;Beck and Knox, 1971) and ethyl acetate-light petroleum (3 : 1 and 1 : 1) (Guggolz et al, 1961). Isoflavones from the heartwood extracts of Cladrastis lutea have been chromatographed with chloroform-methanol (4 : 1) and ether (Shamma and Stiver, 1969).…”

The Oestrogenic Activity of Red Clover Isoflavones and Some of Their Degradation Products

“…Another closely related isoflavone, daidzein (IV), previously found in soya bean as the glucoside, has also recently been detected in red clover and other forages (Guggolz, Livingston & Bickoff, 1961;Wong, 1962 The oestrogenic activities of these isoflavones have been compared with that of diethylstilboestrol by Cheng, Yoder, Story & Burroughs (1954). They found daidzein to be somewhat more active than genistein and biochanin A, and formononetin to have a slight activity.…”

“…Also in view of the findings that formononetin and biochanin A are the predominant isoflavones in red clover (Guggolz et al 1961 ;Wong, 1962) the question of the activity, if any, of formononetin becomes of importance. In this paper the results of a com¬ parative study of the activity of the isoflavones at three dose levels using the mouse uterine weight bioassay are presented, together with the results of a comparison of the oestrogenic activity of a sample of red clover and biochanin A.…”