Simultaneous determination of phytoestrogens in dietary supplements by high-performance liquid chromatography

Abstract

The bio-active phytoestrogen compounds (puerarin, daidzin, glycitin, genistin, miroestrol, daidzein, glycitein, genistein) in dietary supplements were extracted by ultrasonic method with methanol solvent in 40oC. The analysis procedure was carried out on HPLC Alliance e2695 (Waters) system, with RP-C18 Reliant (25 mm × 4.6 mm; 5µm) column, at 30oC. Phytoestrogens were separated by using gradient elution with a mobile phase consisting 0.1% (v/v) phosphoric aqueous solution and methanol in 45 minutes. The method was validated by determining its specificity, linear range, limit of detection, limit of quantification, repeatability and accuracy. This method was applied succesfully to determine content of Phytoestrogens in commercial dietary supplement products.