Extraction and quantification of daidzein and genistein in food

A simple analytical method has been developed for routine quantification of a broad range of concentrations of the isoflavones daidzein and genistein in food. The synthetic glucosides daidzin and genistin were used as internal standards, combined with each food prior to extraction. The recovery of the aglycones daidzein and genistein from these internal standards were used to ensure the completeness of the extraction and aid quantification of isoflavones from the food. Hydrolytic enzymes from Aspergillus niger were used, in aqueous buffer, to liberate daidzein and genistein from their respective glycosides. The aglycone isoflavones were partitioned from the aqueous buffer into ethyl acetate. After evaporation of the ethyl acetate under nitrogen, the isoflavones were derivatized with N-tert-(butyldimethylsilyl)-N-methyltrifluoroacetamide and quantified by comparison with authentic synthetic standards using gas chromatography-mass spectrometry in selected ion mode. The isoflavone content of a stock soy flour was determined, using 36 separate assays, to be 1.05 mg daidzein and 1.11 mg genistein per gram of freeze-dried food, and the interassay coefficient of variation was 2.7 and 4.7, respectively.