Quantitative HPLC analysis of isoflavonoids and bioassays of the farm-grow White Kwao Krua Pueraria mirifica

HPLC isoflavonoid analysis of 5 clones of Pueraria mirifica tubers in a field trial at Ratchaburi province collected in 3 seasons revealed that the 5 isoflavonoids; puerarin, daidzin, genistin, daidzein and genistein were varied. Correlation analysis of the amount of rain and temperature change with tuberous isoflavonoid contents indicated the correlation between genistin, the ratio of glycoside/aglycoside, the ratio of aglycoside and glycoside/puerarin with the rainfall amount in rainy season and winter. In rainy season, the ratio of aglycoside/glycoside was negatively correlated with the rainfall amount. Whereas, temperature change had no correlation with isoflavonoid contents. We can conclude that the rainfall amount had influence on isoflavonoid storage in P. mirifica tubers. The 5 plants exhibited different proliferation effects on MCF-7. PM-I, PM-II and PM-V collected in summer and PM-III collected in winter exhibited proliferation while PM-II collected in winter exhibited antiproliferation effect. MCF-7 proliferation assay was much influenced by the presence of S9 mixture which could increase the proliferation response. The studies in ovariectomized rats treated with 100 and 1,000 mg/kg body weight (BW) of plant powder dissolved in 0.7 ml of distilled water, compared to rats fed with 0.7 ml of distilled water only and rats injected with a single dose of 17β-estradiol. The experiments were set into 3 phases, pre-treatment period for 14 days, treatment period for 14 days and post-treatment period for 7 days, with 5 parameters. From the first day of appearance of vaginal cornified cell in the treatment period and the total day of appearance of cornified cells in the treatment period and post-treatment period including, it was found that the estrogenic activity of P. mirifica was a dose dependent. The vaginal cornification in rats treated with1000 mg/kg BW occurred faster and longer than those of rats treated with 100 mg/kg BW. Differentiation of vaginal cells after P. mirifica treatment agreed with the increased uterine weight. From the 5 analyzed parameters, Samples collected in summer and winter showed stronger estrogenic activity than in rainy season. PM-III showed stronger estrogenic activity than other clones. Isoflavonoid activity, especially aglycoside (daidzein and genistein) and glycoside (daidzin and genistin) in differ seasonal collected P. mirifica had influence on estrogenic activities. The conclusion from this study is the physical factors and plant genetics exhibited influences on the plant estrogenic activities. Therefore, the right harvest season and plant clone selection for high estrogenic activity could be applied as parameters to establish high quality raw materials for industry demand.