This paper focuses on the evaluation of water quality variations in Hirfanlı Water Reservoir, which is one of the most important water resources in Turkey, through EO-1 (Earth Observing-1) Advanced Land Imager (ALI) multispectral data and real-time field sampling. The study was materialized in 20 different sampling points during the overpass of the EO-1 ALI sensor over the study area. A multi-linear regression technique was used to explore the relationships between radiometrically corrected EO-1 ALI image data and water quality parameters: chlorophyll a, turbidity, and suspended solids. The retrieved and verified results show that the measured and estimated values of water quality parameters are in good agreement (R (2) >0.93). The resulting thematic maps derived from EO-1 multispectral data for chlorophyll a, turbidity, and suspended solids show the spatial distribution of the water quality parameters. The results indicate that the reservoir has average nutrient values. Furthermore, chlorophyll a, turbidity, and suspended solids values increased at the upstream reservoir and shallow coast of the Hirfanlı Water Reservoir.