A novel method for the rapid and sensitive analysis of 1-hydroxypyrene (1-OHP) in human urine has been developed that uses a resonance light scattering (RLS) technique. The assay was based on the interaction of ethyl violet (EV) with 1-hydroxypyrene to form an ion-associate complex, which resulted in the enhancement of RLS intensity and the appearance of new RLS spectra. In the presence of anionic surfactant, the maximum RLS peak of the system was located at 396 nm at pH 8.0. Under the optimum conditions, it was found that the enhanced RLS intensity was directly proportional to the concentration of 1-hydroxypyrene in the range of 4.0 -982 μg l -1 . The detection limit was 1.2 μg l -1 and the recoveries of 1-hydroxypyrene were 92.8 -102.3% (n = 6). The proposed method was successfully applied to the analysis of human urine samples. The results of 1-hydroxypyrene were in agreement with those obtained by the method of highperformance liquid chromatography.