Abstract: A high performance liquid chromatography tandem mass spectrometric (HPLC-MS/MS) method was established to simultaneously detect colistin sulfuric (CS), bacitracin (BTC) and virginiamycin M₁ (VGMM₁) in aquatic products. We extracted the samples with methanol-0.1% formicacid(V : V=2 : 5), removed their protein with 4% TCA acetonitril, removed fat with acetonitrile saturated n-hexane, and then cleaned them up with OASiS HLB solid phase extraction (SPE) column. Next, we detected the analytes by HPLC-MS/MS under the selected reaction monitoring mode and quantified them by external standard method. Results show that CS and BTC have good linearity between the peak areas in concentrations ranged 0.01~10.00 mg · L^-1, VGMM₁ as well in concentrations ranged 0.002~2.000 mg · L^-1, and the correlation coefficients (R²) are all more than 0.995. The detection limits of CS, BTC and VGMM₁ are 10 μg · kg^-1, 10 μg · kg^-1and 2 μg · kg^-1 respectively, and the quantification limits are 20 μg · kg^-1, 20 μg · kg^-1 and 4 μg · kg^-1respectively. Three samples in three different concentration levels are chosen for recovery, getting a 72.3%~103.9% average recovery rate, and a relative standard deviations varying from 1.10%~10.92%. In conclusion, method proposed has advantages of simplicity, sensitivity and repeatability, which provide technical support for the detection of these 3 kinds of drug residues in aquatic products.