Abstract: A method based on high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was developed for simultaneous determination of five yellow industrial dyes. The separations were performed with an Agilent ODS C18 column at a flow rate of 0.3 mL/min. The mobile phase was 5 mmol/L ammonium acetate (containing 0.1% formic acid)-acetonitrile (3:2, v/v). Under the optimized detection conditions, the linear ranges for Chrysoidine G and Basic Yellow 2 were 5.0~80.0 mg/L, and for Acid Orange I, Acid orangeII and Acid Yellow 36 were 10.0~160.0 μg/L. The limits of quantification for Chrysoidine G, Basic Yellow 2, Acid Orange I, Acid OrangeII and Acid Yellow 36 were 20, 20, 40, 40 and 40 ng/g, respectively. The relative standard deviations of reproducibility of this method for retention time and peak area were no more than 0.50% and 2.14%, respectively. The method was applied to determine the recoveries of the above five dyes in chicken, bean products and yellow croaker were between 79.8%~95.2% with satisfactory results.