Objective To investigate the oxidation of acenaphthene (Ace), a polycyclic aromatic hydrocarbon (PAH) with a saturated C-C bond by ozone and to characterize the intermediate products of ozonation. Methods Ozone was generated from filtered dry oxygen by an ozone generator and continually bubbled into a reactor containing 1g/L Ace dissolved in an acetonitrile/water solvent mixture (90/10, v/v) at a rate of 0.5 mg/s. HPLC was used to analyze the Ace concentration. Total organic carbon (TOC) was used to measure the amount of water soluble organic compounds. GC-MS was used to identify the ozonized products. Oxygen uptake rate (OUR) of activated sludge was used to characterize the biodegradability of ozonized products. Results During the ozonation process, Ace was degraded, new organic compounds were produced and these intermediate products were difficult mineralize by ozone, with increasing TOC of soluble organics. The ozonized products were degraded by activated sludge more easily than Ace. Conclusion Ozonation decomposes the Ace and improves its biodegradability. The ozonation combined with biological treatment is probably an efficient and economical way to mineralize acenaphthene in wastewater.