A method for the determination of four polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]anthracene, chrysene, benzo[b]fluoranthene and benzo[a]pyrene in olive oil by high performance liquid chromatography-fluorescent detection (HPLC-FLD) was developed. The olive oil samples were diluted with isopropanol and cleaned-up by a solid phase extraction cartridge with π-π specificity. Critical isomers were separated by gradient elution with water-acetonitrile on an Agilent ZORBAX Eclipse PAH column (100 mm×2.1 mm, 1.8 μ m). The calibration was performed by the use of matrix-matched calibration standards. The results showed that there were good linear relationships for the four PAHs in the range of 2.4-40 μ g/L with correlation coefficients of 0.9990-0.9999. The limits of quantification (LOQs) ranged from 0.147 to 0.413 μ g/L, and the recoveries of the spiked samples ranged from 95.5% to 103.2%. The intra-day and inter-day precisions were in the ranges of 0.10%-1.69% and 2.48%-2.93% (n=5), respectively. The method has the characteristics of high sensitivity, low detection limit and good precision, and is suitable for rapid and accurate quantitative detection of the four PAHs in olive oil.