High-performance liquid chromatographic assay of isoflavonoids and coumestrol from human urine

A rapid, sensitive and precise diode-array reversed-phase HPLC method was developed for human urine analysis of the most common dietary isoflavones daidzein, genistein, formononetin and biochanin-A, their mammalian metabolites equol and O-desmethylangolensin, and of coumestrol, another commonly occurring phytoestrogen. Analytes were isolated and concentrated by solid-phase extraction and separated by HPLC followed by identification through retention times and UV scans, and in the case of coumestrol additionally by fluorometric response. This method was applied to monitor changes in urinary excretion of these analytes after challenge with soybeans and was evaluated for precision and recovery of analytes.