An optimized method for the determination of benzopyrene in vegetable oil by molecular imprinting solid phase extraction with ultra-high performance liquid chromatography was established. During the experiment, through the investigation of different solid-phase extraction purification efficiency, optimal detection wavelength, mobile phase system and chromatographic column use, the results showed that the molecularly imprinted solid phase extraction purification column was more suitable for pretreatment extraction and purification of vegetable oil. The optimal detection the excitation wavelength and the emission wavelength were 365 nm and 410 nm, respectively. Under these conditions, the benzopyrene chromatographic peak had high response value and signal-to-noise ratio. Under the mobile phase system of 90% acetonitrile-10% water, the target peak had a good separation effect. Using 150 mm short columns can effectively improve work efficiency. Benzopyrene had a good linear relationship in the concentration range of 0.3~20 µg/L, and the correlation coefficient was greater than 0.999 9. The detection limit and quantitative limit of benzopyrene determination method were 0.1 µg/kg and 0.3 µg/kg, respectively. The average recoveries of three added concentrations were 87.6%~95.9%, and the relative standard deviations were 1.08%~2.16%. In the actual sample detection, the detection rate of benzopyrene, the exceeding rate and the qualified rate were 39.0%, 2.4%, and was 97.6%, respectively. This optimization method is sensitive, convenient, rapid and accurate, and can be used for batch detection of benzopyrene in vegetable oil and analysis of pollution monitoring.