A novel fluorescent reagent o-vanillin-8-aminoquinoline (OVAQ) was synthesized, and its infrared spectrum, elemental analysis and acid-base dissociation constants were obtained. The fluorescent reaction of this reagent with Cr (III) was studied. In acetonitrile-water (1:1, (v/v)) medium of pH 6.00, Cr (III) could react with fluorescent reagent OVAQ (λex/em=280/314 nm) to form a 1:1 non-fluorescent complex. The linear range of the spectrofluorimetric method proposed was from 8.2 to 130g l−1, and the detection limit was 2.5g l−1. The interferences of 25 foreign ions were also studied. This method could be easily performed and was successfully applied to the determination of Cr(III) and total chromium in domestic and industrial waste water samples.