Abstract
A reliable and sensitive method for quantification of daidzein and genistein by HPLC with coulometric electrode array detection is presented using bisphenol A as internal standard. Acid hydrolysis during extraction of foods allows the quantitative determination of total phytoestrogens as aglycones. The substances are separated on a reversed phase column (Hypersil® Elite C-18), eluted with methanol/acetonitrile/50 mM sodium acetate pH 4.8 (40/ 20/40, v/v/v) and detected in a coulometric electrode array detector using an oxidative detection mode (+390 to +810 mV in increments of 60 mV, vs palladium reference electrode). Phytoestrogen levels from several food items were determined. High levels of daidzein (819 mg/kg) and genistein (960 mg/kg) were found in soy products whereas biochanin A could not be detected in any of these food samples. The recovery depends on the kind of food and was found to be between 72 and 94% for daidzein and genistein.
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