“NANO BASED ENHANCEMENT AND RP-HPLC QUANTIFICATION OF DAIDZEIN, GENISTEIN AND FORMONONETIN IN CAYRATIA TRIFOLIA”- A BONE-HEALING PLANT OF SEMI-ARID LANDS

A fast, sensitive and precise method is presented for the efficient extraction and quantification of daidzein, genistein and formononetin (Phytoestrogens) in in vivo and in vitro samples from Cayratia trifolia by diode array reversed phase (RP) Agilent 1200 infinity High Performance Liquid Chromatography (HPLC) system using phytoestrogen as external standard. Samples were eluted using a C-18 reverse phase column under gradient (0.025% Ortho-phosphoric acid in Milli-Q: Acetonitrile) condition in less than 30 min. Standard daidzein, genistein and formononetin were eluted at retention time (Rt) 14.222, 19.289 and 23.722 min, respectively. Among all samples (conforming these phytoestrogens), callus showed the maximum amount of all phytoestrogens especially formononetin (2.654×10-1 mg/ml) while stem showed formononetin (3.84 ×10-1 mg/ml) only. The present study also demonstrates an effect of nano-ZnO and Fe3O4 particles on the growth of in vitro grown callus. It was found that at certain optimum concentration, 0.025 ml/l and 0.01 ml/l respectively displayed good growth over control, and beyond that, retardation in growth was observed. The result indicated that the method is suitable for determination of phytoestrogens in Cayratia trifolia using advanced efficient 1200 infinity RP-HPLC. This is the first report of isolation, quantification and enhancement of phytoestrogens from Cayratia trifolia by RP-HPLC.