Abstract: A method for detecting the estrogens estriol, 17β-estradiol, ethinyl estradiol, and bisphenol A in livestock dung was established by the combination of ultrasonic extraction (UE), solid phase extraction (SPE) purification, and high performance liquid chromatography (HPLC) with fluorescence detector (FLD). The dung samples were extracted with ethyl acetate ultrasonication for 30 min, and purified with C₁₈ solid phase extraction column and related solvents. The test four estrogens in the dung samples were isolated with Inertsil ODS-SP-C₁₈ reversedphase columns (150 mm×4.6 mm, 5 μm), and the isolated estrogens were detected with HPLC/FLD. The mobile phase of HPLC for the detection was methanol/acetonitrile/water (volume ratio of 20:30:50), with a flow rate of 0.8 mL·min^-1. The excitation and emission wavelengths of FLD were 280 and 310 nm, respectively, the HPLC column temperature was 40 ℃, and the injection volume was 20 μL. Good linearity (correlation coefficient greater than 0.9995) was observed by the HPLC/FLD detection when the test four estrogens concentrations were in the range of 1.00-1000.00 μg·L^-1. The detection limit of estriol, bisphenol A, 17β-estradiol, and ethinyl estradiol was 3.35, 5.01, 2.13, and 1.12 μg·kg^-1, respectively. When the added estrogens concentrations of pig, cow, and chicken dung samples were 0.05, 0.40, and, 1.00 μg·kg^-1, the average recovery of the four estrogens was 75.1%-91.1%, 78.4%-117.0%, and 78.6%-97.8%, respectively, with the relatively standard deviations (RSD, n=6) all less than 6%. By adopting the established SPEHPLC/FLD method to detect the estrogens in real pig, cow, and chicken dung samples from parts of the largescale livestock raising farms in Nanjing of East China, the detection reproducibility was high, and the detection limit was low, being available and effective for the detection of the estrogens in livestock dung.