The development of a real-time monitoring tool for the estimation of water quality is essential for efficient management of river pollution in urban areas. The Gap River in Korea is a typical urban river, which is affected by the effluent of a wastewater treatment plant (WWTP) and various anthropogenic activities. In this study, fluorescence excitation-emission matrices (EEM) with parallel factor analysis (PARAFAC) and UV absorption values at 220 nm and 254 nm were applied to evaluate the estimation capabilities for biochemical oxygen demand (BOD), chemical oxygen demand (COD), and total nitrogen (TN) concentrations of the river samples. Three components were successfully identified by the PARAFAC modeling from the fluorescence EEM data, in which each fluorophore group represents microbial humic-like (C1), terrestrial humic-like organic substances (C2), and protein-like organic substances (C3), and UV absorption indices (UV220
), and the score values of the three PARAFAC components were selected as the estimation parameters for the nitrogen and the organic pollution of the river samples. Among the selected indices, UV220
, C3 and C1 exhibited the highest correlation coefficients with BOD, COD, and TN concentrations, respectively. Multiple regression analysis using UV220
and C3 demonstrated the enhancement of the prediction capability for TN.