A multi-residue method was developed for the simultaneous determination of 8 estrogens,5 androgens,6 progesterones,8 glucocorticoids and 3 chloramphenicols in fish using QuEChERS sample preparation and liquid chromatography-tandem mass spectrometry（LC-MS/MS）.The homogenized sample was dispersed by water,then the analytes were extracted with acetonitrile.The separation were performed on a ZORBAX Extend-C18 column（100 mm×2.1 mm，3.5 μm） by gradient elution after dispersive-solid-phase extraction（d-SPE） purification.The qualitative and quantitative analysis of 30 analytes were operated by electrospray ionization mass spectrometry under the positive or negative mode using multiple reaction monitoring（MRM）.Acetonitrile and water were used as the mobile phase in the positive mode.Acetonitrile and 0.1% ammonia hydroxide were used as the mobile phase in the negative mode.The correlation coefficients of calibration curves were over 0.99 in the corresponding concentration range.The average recoveries of the 30 analytes at three spiked concentration levels varied from 63% to 118% with relative standard deviations（RSDs） of 3.8%-18.2%.The limits of detection（LOD,S/N≥3） and quantitation（LOQ,S/N≥10） were 0.03-1.6 μg/kg and 0.1-5.0 μg/kg,respectively.The real sample tests showed that the proposed method was simple,rapid,sensitive,reliable and cost-effective,and was suitable for the simultaneous determination of hormones and chloramphenicols in fish samples.