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Large volumes of oil sands process-affected waters (OSPW) are produced during the extraction of bitumen from oil sand. There are approximately 10 9 m 3 of OSPW currently being stored in settling basins on oil sands mining sites in Northern Alberta. Developers plan to create artificial lakes with OSPW and it is expected that this water may eventually enter the environment. This study was conducted in order to determine if synchronous fluorescence spectroscopy (SFS) could detect OSPW contamination in water systems. Water samples collected from ponds containing OSPW and selected sites in the Alberta oil sands region were evaluated using SFS with an offset value of 18nm. OSPW ponds consistently displayed a minor peak at 282.5nm and a broad major peak ranging between 320 and 340nm. Water from reference sites within the oil sands region had little fluorescence at 282.5nm but greater fluorescence beyond 345nm. Naphthenic acids are the major toxic component of OSPW. Both a commercial naphthenic acid and a naphthenic acid extract prepared from OSPW had similar fluorescent spectra with peaks at 280nm and 320nm and minor shoulders at approximately 303 and 331nm. The presence of aromatic acids closely associated with the naphthenic acids may be responsible for unique fluorescence at 320–340nm. SFS is proposed to be a simple and fast method to monitor the release of OSPW into ground and surface waters in the oil sands region.