In order to investigate the potential risk of anesthetic application in fish transportation, the method was developed to determine residue of eugenol in water. The target compound in water sample was extracted using ethyl acetate, and then conducting the qualitative and quantitative determination by using gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). As shown, within 1.00~200.00 μg / L concentration range of the standard curve equation y = 14.65 x - 7.23, correlation coefficient (r) is 0.999 4, the linear fit is good. While spiked concentration from 5.00 to 100.00 μg/L, the recoveries of eugenol ranged from 98.6% to 104%, and the relative standard deviation (RSD) were ranged from 2.46% to 5.48%. The limit of detection (LOD) and limit of quantification (LOQ) values were 0.21 and 0.68 μg/L, respectively. It can be concluded that the method is accurate, simple, fast and qualified to detect eugenol residue in water during fish transporting.［Chinese Fishery Quality and Standards, 2014, 4(4):49-55］