A reversed-phase isocratic elution high-performance liquid chromatography method coupled with fluorescence detection has been developed to determine urea concentration via online postcolumn derivatization. Swimming pool water samples were filtered through 0.20 μm syringe filters. When the temperature of reaction coil was 40°C, urea was derivatized well with xanthydrol methanol solution (0.1 g/L) containing 0.50% hydrochloric acid with a flow rate of 0.20 mL/min. Successful separation was achieved by using Shim-pack VP-ODS C18 (250 mm × 4.6 mm, 5 μm) column, with a mobile phase containing phosphoric acid solution (0.01 mol/L) at a flow rate of 0.80 mL/min. Retention time and external standard method were used for qualitative and quantitative urea analysis, respectively. Under the established conditions, the limit of detection, linear range, correlation coefficient, recovery and relative standard deviation was 0.09 mg/L, 1.0-100.0 mg/L, 0.9998, 87.0-105.3% and 0.95-4.8%, respectively. Ammonia, thiourea and trichloroisocyanuric acid did not interfere with urea analysis. The method showed satisfactory results with high precision, accuracy, recovery, as well as sensitivity, for the determination of urea in swimming pool water.