Simultaneous Determination of Isoflavones and Bisphenol A in Rat Serum by High-performance Liquid Chromatography Coupled with Coulometric Array Detection

A method for simultaneous detection and quantification is presented to determine the presence of isoflavones and bisphenol A in a biological sample. A coulometric array detector was used with reversed-phase high-performance liquid chromatography (HPLC). Daidzein (1), glycitein (2), genistein (3) and their glucoside conjugates, daidzin (4), glycitin (5) and genistin (6), were measured as phytochemicals. Also assayed here was equol (7), a metabolite from compound 1, and bisphenol A (8), an industrial chemical that acts as an endocrine disrupter. All chemicals were simultaneously detected by using a 600-mV single detection voltage with high efficacy. A mixture of 1, 3and 8was orally administered to rats, and the levels of these three chemicals in the serum were clearly increased after a 4 kU β-glucuronidase treatment. The levels of compounds 1and 3in the serum were detected at 1665 and 2040 ng/ml, while 8was at a low level of 417 ng/ml. Compound 7in the serum was not detected until after enzymatic hydrolysis (72 ng/ml). These results suggest that this analytical method would be useful for metabolic and pharmacokinetic studies on isoflavones and bisphenol A.