A method was developed for the determination of chloramphenicol in propolis by high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS).The flavonoid was removed with NaOH after the dissolution of the sample with methyl tert-butyl ether（MTBE).The solubility of chloramphenicol in MTBE was decreased with n-hexane，and then chloramphenicol was extracted with sodium acetate buffer.After the extraction was transferred to alkaline solution，ethyl acetate was used to extract chloramphenicol.The supernatant was evaporated,and re-dissolved with water for detection.The separation of chloramphenicol was performed on an Atlantis T3 column with methanol and water（65∶35，by volume) as mobile phases，and the detection of chloramphenicol was carried out by tandem mass spectrometry in negative electrospray ionization（ESI-) and multiple reaction monitoring（MRM) mode.The quantification was performed by the internal standard method.The calibration curve showed a good linearity in the range of 0.1-5.0 μg/L.The limits of quantitation（S/N≥10) were 0.3 μg/kg.The recoveries at three spiked levels were in the range of 97.3%-103%with relative standard deviations of 4.8%-6.4%.With the advantages of sensitivity，strong anti-interference capability，good recovery and repeatability，the method was suitable for the monitoring of veterinary residue analysis