Benzo [a] pyrene and benzo [e] pyrene in food was extracted and purified by refrigerated centrifugation with dimethyl sulfoxide as solvent and analyzed by HPLC-fluorescence detection method. This method can replace the traditional solid-phase extraction column purification method, which is faster, more efficient and accurate in the detection of benzo [a] pyrene and benzo [e] pyrene content. Both target object showed a good linear relationship within range of 0.5～1000ng/mL, when addition was 1～100ng/g, the recovery rate was 89.0%～91.3%, and the relative standard deviation was 1.19%～1.29%; the detection limit was 0.01ng/mL, vegetable fat detection limit was 0.05ng/g, meat and animal fat detection limit was 0.02 ng/g. This method is faster and more convenient in the detection of benzo [a] pyrene and benzo [e] pyrene in animal, vegetable fat and meat products, which is suitable for batch detection.