Abstract: A high performance liquid chromatography method with fluorescence detector was developed for determination of anionic surfactant in vegetable oil and waste oil. The oil samples were dissolved with hexane and extracted with water. A Waters Atlantis T3 column was used with methanol as the mobile phase at a flow rate of 2.0 mL/min. The column temperature was 25℃ and the Fluorescence detection wavelength was λex/em=223/280 nm. Under the optimized conditions, the calibration curve shows good linearity in the range of 1~ 100 ug/mL of anionic surfactant, Y = 222626 X-329085 (r=0.9996，n=6). The recovery and the relative standard deviation of method were 87.56%~93.29% and 1.51%～2.63%，respectively. The method is simple and accurate, and can be used for the quality control of anionic surfactant in vegetable oil. We found that the content of anionic surfactant in common vegetable oil is lower detection limit. In order to prevent waste oil from mixing in vegetable oil, we suggest that the limit standard of anionic surfactant in vegetable oils must be developed.