The method for the determination of three chloramphenicols including chloramphenicol (CAP)，thiamphenicol (TAP) and florfenicol (FF) in aquatic products was developed by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Deuterium (d5-CAP) was used as internal standard instead of CAP, and was added to the sample before extraction with ethyl acetate. The sample was extracted with ethyl acetate to transfer CAP, TAP and FF into the organic phase. The extract liquid which consequently dried by blowing with nitrogen was dissolved with water and defatted with hexane. The mass spectrometer was operated in the negative ion mode using select reaction monitoring for qualitative and quantitative analysis of these compounds at the same time. The preliminary treatment was predigested，and no solid phase extraction (SPE) procedure was adopted. The advantages of the method are simple operation，less organic chemicals consumed，and shorter operation time. The limit of detection (LOD) for CAP was 0.01 μg/kg and 0.03 μg/kg for TAP and FF.