To establish an improved solid phase extraction-high performance liquid chromatography method with fluorescence determination for the detection of benzopyrene (BP) in cooking oil.Methods Cooking oil samples were prepared by neutral alumina solid phase extraction (SPE) column. This study investigated the effect of grease removal and recovery of BP as a function of water content in the neutral alumina packing. Neutral alumina packing with 6% water content was found to be the best solid phase extraction packing for BP pre-treatment. In the HPLC method, the HPLC column used was Ultimate XB-C₁₈ (250 mm×4.6 mm, 5 μm) column from Welch Materials, Inc. The mobile phase was acetonitrile-water (88∶12, V/V) with a flow rate of 1.0 ml/min, the column temperature was 30 ℃, and the injection volume was 10 μl.Results The limit of determination was 0.5 ng/g (S/N=3), the linear range was 0.5-100 ng/ml, the recoveries were 91.2%-101.0% and the relative standard deviations were 2.2%-2.9%.Conclusion The method is highly sensitive with good selectivity and accurate quantification, which is suitable for BP determination in cooking oil.