Abstract: A method for analysis of chloramphenicol residue in bee pollen was developed. Sample was extracted using accelerated solvent extraction with ethyl acetate as extraction reagent and purified by Silica solid phase.Mass spectrometer was operated in the negative ion mode using select reaction monitoring. Isotope internal standard was used for quantitative analysis. The linearity of the calibration curve was from 0.5 to 10 ng/m L with correlation coefficient was 0.9936. Recovery rates for chloramphenicol in a negative bee pollen sample spiked at three levels were between 88.3% and 110.0%,with RSD range of 5.3% ⁸.9%. The limit of detection in the method was 0.1 μg/kg. This method proved to be of high efficiency,small amount of reagent,good selectivity and high sensitivity.