This study aimed to measure phytoestrogen contents in health functional foods (HFFs) to alleviate menopausal symptoms that are commercially available in South Korea. A sensitive and selective method using gas chromatography–mass spectrometry (GC-MS) for the monitoring of 21 phytoestrogens was developed and fully validated. Phytoestrogens were extracted by liquid–liquid extraction using methyl tert-butylether as the solvent. Food samples were enzymatically hydrolyzed with β-glucuronidase/arylsulfatase. Before GC-MS, the analytes were derivatized to their trimethylsilyl derivatives by incubation with N,O-bis(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane for 30 min at 80°C. The calibration curves showed good linearity (r 2 > 0.99) in the range of interest for each analyte. The limits of detection and quantification of the 21 phytoestrogens were in the ranges of 0.5–20 and 1–50 ng/mL, respectively. The intra- and inter-day precisions were all below 15.7%. These multivariate procedures were efficient in determining the optimal conditions using peak heights as responses.
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