The purpose of this research is to develop an UPLC/QTOFMS method for the rapid and simultaneous determination of six estrogens (bis-phenol A, 17α-ethinylestradiol, 17β-estradiol, estrone, dienestrol, estriol) in animal-derived food. The method aimed to be simple to operate with good sensitivity. Sample preparation involved matrix solid-phase dispersion extraction (MSPD) using acetonitrile in C18 and Florisil packings. The pretreatment method of MSPD was optimized to eliminate the complications of the classical pretreatment. Estrogens were qualitatively and quantitatively analyzed using a C18 reversed-phase UPLC/QTOFMS in target MS/MS mode. The six estrogens were successfully separated within 4 minutes with the optimized method of one simple elution with water/acetonitrile 52/48 (v/v). The limits of detection were 0.11–0.81 μg L−1, based on an S/N = 3. The limits of quantification were 0.61–2.20 μg L−1, based on an S/N = 10. The recoveries were determined to be in the range of 59.6–128.5%, with a relative standard deviation (RSD) between 1.2% and 8.0%. Milk and meat samples were analyzed, and the presence of the six estrogens was determined. The detected rate of real samples shows that the high-fat food tends to be the source of estrogens among animal-divided food. An UPLC/QTOFMS method was developed to determine six estrogens in animal-derived food and was successfully applied to real samples, and it showed good sensitivity and simple operation.
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