The Research Of Red Clover Isoflavones Transforming Into Genistein And Daidzein

Experiment studied the demethylation reaction of formononetin transforming into daidzein. The results show that the optimum conditions were KI-H3PO4 as the reaction medium, time was 4h, temperature was 100℃, liquid ratio was 1:15, saturatione was 75%, the yield was 87.03%. The optimum conditions were KBr-H3PO4 as the reaction medium, time was 5h, temperature was 120℃, liquid ratio was 1:45, saturation was 75%, the yield was 89.42%.Experiment studied the demethylation reaction of biochanin A transforming into genistein. The results show that the optimum conditions were KI-H₃PO₄ as the reaction medium, time was 5h, temperature was 100℃, liquid ratio was 1:45, saturatione was 75%, the yield was 88.51%. The optimum conditions were KBr-H₃PO₄ as the reaction medium, time was 5h, temperature was 120℃, liquid ratio was 1:45, saturation was 100%, the yield was 85.98%.The products were isolated and purified by TLC. The best separation condition was petroleum ether:ethyl acetate (1:1) as developing solvent. The products were isolated and purified by column chromatography, then the puritys of them were 98.26% and 98.51% ( detection was external standard method of HPLC ). The products were synthesized and characterized with the methods of IR spectra and NMR spectrum etc.Extracted red clover by ultrasonic vibration, which the amount of ethanol was 10 times (V/W), time was 90 min, temperature was 60℃, and extraction rate was 74.93%. Purified extracts by AB-8 macroporous resin, isolated the main components of formononetin and biochanin A, their purities were 71.52% and 80.74%. By demethylation methods respectively, production rates of daidzein and genistein were 68.33% and 65.01%.The antioxidant activity of transformation products of genistein and daidzein was studied by means of scavenging ability of superoxide free radicals, removing the ability of DPPH, anti-linoleic acid capacity and reducing power. The results showed that capacity of scavenging superoxide radical of them increased with increasing concentration. At the same concentration, the removal of genistein was higher than daidzein.The antibacterial activity of the transformation products of genistein and daidzein was studied by determinating the inhibition of Escherichia coli,Sporolactobacillus Kitahara,proteusbacillus vulgaris and Staphyloccocus aureus Rosenbach. The results showed that they had a weak inhibitory effect on tested bacterias in 0.030mg/mL, which minimum inhibitory concentration (MIC) of them was 0.030mg/mL. Both of them had antibacterial effects on test bacterias, with increasing the concentration, inhibitory effects were obvious. Their antimicrobial activity was that Staphyloccocus aureus Rosenbach> Escherichia coli> proteusbacillus vulgaris> Sporolactobacillus Kitahara and the inhibitory effect of Staphyloccocus aureus Rosenbach was the strongest.