This work helps to identify the source of pollution in water and characterize the water quality which is essential to water management for sustainable development. Therefore, the main objective of this work is to evaluate the spatial distribution of the water quality of Ratuwa river and its tributaries. The water samples were collected from six discrete sampling locations and fifteen parameters were tested using respective well-calibrated equipment and standard APHA methods. The physicochemical analysis, water quality index, and correlation matrix method were employed to evaluate the spatial variation of the water quality of Ratuwa river. Turbidity was the most polluting factor in river water. The results showed the spatial variation of the water quality index (WQI) from 39.3 to 70.5, which fell in the range of “good” to “poor” water quality status. None of the water samples was either “excellent” or “unsuitable for drinking.” The water quality was “Poor” upstream and downstream of Ratuwa river due to the high value of turbidity. Chaju river was found to have unpolluted whereas Dipeni river was slightly polluted due to domestic and municipal wastes. Hence, the deterioration of water quality can be attributed to natural and anthropogenic sources.