A simple and rapid analytical method was developed for the simultaneous determination of albuterol, bisphenol A, metandren in culture water using ultra-performance liquid chromatography-electrospray tandem mass spectrometry. The hormone residues were extracted from the samples and cleaned-up by ASPE, then analyzed using LC-MS/MS in multiple reaction monitoring mode via positive electrospray ionization with BEHC18 column. The linearity of all hormone were good in the concentration range of 10-100 ug/L. The recoveries of all hormones were in the range of 80%-120%. And the limits of quantification were no more than 1 ug/L.