Objective To establish a detection method for 9 yellow colorants in fish tissues by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods Sample was extracted by ammonia methanol solution and formic acid water methanol solution, the supernatant was evaporated and redissolved, the fat was removed by n-hexane, determinated by UPLC-MS/MS method after centrifugation, and quantification was based on external calibration. Results The result showed good linearity in the ranges of 2-30 μg/kg for tartrazine, sunset yellow and quinoline yellow, 0.2-3.0 μg/kg for acid orange Ⅱ, basic orange 2, basic orange 21, basic orange 22, thioflavin T and auramine O, and the limits of detection were 2 μg/kg and 0.2 μg/kg. The recoveries were 80.03%-107.06% and the relative standard deviations(n=6)were 1.2%-6.6%. Conclusion The method can be applied to determine 9 yellow colorants in fish tissues.