Development Of RP-HPLC For Daidzein And Genistein And Detection Of Daidzein And Genistein Concentration In Organism And Egg Of Poultry

1.The development of HPLC for daidzein and genisteinA reverse-phase high performance liquid chromatographic method was established for the determination of daidzein and genistein. Waters Spherisorb ODS2 column (4.6mm i.d.× 150mm, 5μm) was used with a mixture of methanol-water (60:40,v/v) as the mobile phase and a detector set at 254nm. The calibration curve was linear(r=0.9984,p<0.01) in the range of 0.05μg/ml-20μg/ml of dadizein and was linear(r=0.9978,p<0.01) in the range of 0.05μg/ml-10μg/ml of genistein. The recoveries of dadidzein and genistein were 91.72%-98.56% and 96.80%-98.60% respectively with the intra-day and inter-day RSDs of 1.30%, 5.30% and 1.17%, 3.84%, respectively.2.Effect of daidzein preparation supplemented to basal diets on the performance and daidzein concentration in diet% sera and yolks of laying hens fed ad libitum29 forty-four-week-old laying hens were randomized into 2 groups: basal diet(n=13), basal diet+6mg/kg daidzein(n=16). The blood samples were collected in the 4th day. Compared with control, the feed intake was decreased and laying performance was improved; daidzein concentrations in sera and yolks of laying hens increased by 93.10%(p<0.05) and 68.01%(p<0.05) respectively. Daidzein concentration in feed stuff of laying hens was 18.37μg/g. It indicated that daidzein from feed stuff can be enriched in the yolk.3.Effect of daidzein, genistein and enzyme preparations supplemented to basal diets on the performance and daidzein, genistein concentration in diet, chyme, liver and muscle of chicks192 nine-day-old layer chicks were randomized into 6 groups: basal diet (C group), basal diet plus 3mg/kg daidzein(3Da group), basal diet plus 6mg/kg daidzein(6Da group), basal diet plus 0.05% enzyme preparation+6mg/kg daidzein (Da+E group), basal diet plus 0.05% enzyme preparation(E group) , basal diet plus 6mg/kg genistcin(Ge group). Compared with control, the experiment groups increased chicks body weight gain per day, the feed conversion rate, growth rate and weights of livers and legs. 3Da group increased weight of livers of the second week by 12.22%(p<0.05), 6Da group increased growth rates of the firstweek by 11.01%(p=0.0543), Da+E group increased weight of livers of the second week by 18.98%(p<0.01), Ge group respectively increased weight gain per day and weight of livers of the second week by 8.21%(p<0.1) and 22.56%(p<0.01). 3Da group respectively increased daidzein concentrations in the chyme and liver by 197.16%(p<0.01), 72.84%(p<0.1); 6Da group respectively increased daidzein concentrations in the chyme, liver and muscle by 288.15%(p<0.01),81.48%(p<0.01), 155.97%(p<0.01); Da+E group respectively increased daidzein concentrations in the chyme, liver and muscle by 249.29%(p<0.01), 127.16%(p<0.01), 82.09%(p<0.05), increased genistein concentrations in the chyme and liver by 106.76%(p<0.01),107.07%(p<0.01); E group increased daidzein concentration in muscle by373.88%(p<0.01) , daidzein and genistein concentrations in the chyme and liver had the tendency of increase, however had difference individually; Ge group respectively increased daidzein concentrations in the chyme, liver and muscle by315.64%(p<0.01),72.84%(p<0.05),64.18%(p<0.1),increased genistein concentrations in the chyme and liver by 138.13%(p<0.01),48.48%.In conclusion, daidzein, genistein and enzyme preparation increased chicks performances and daidzein and genistein concentration in the chyme, liver and muscle of chicks. To a certain degree, enzyme preparation can speed daidzein and genistein released from diets and improve daidzein and genistein's absorptivity.