Despite efforts to reduce nonpoint source contamination of water resources, agriculture and urbanization remain primary sources that impair surface water quality. The Scioto River in Ohio, USA has headwaters in land dominated by primarily row-crop agriculture, then flows through metropolitan Columbus, the 14th largest city in the U.S. The river serves as a major drinking water source for the city and is dammed to form two reservoirs. The Scioto River was sampled for major ions, nutrients, and select trace elements weekly to monthly at five locations along changing agricultural and urban land use. Dissolved concentrations of many analytes were highest at the most upstream agricultural location, decreased to downtown Columbus, and increased at the location furthest downstream in agricultural land. The same trend was observed in discharge-weighted yields. Because much of the study area is underlain by carbonate bedrock, correlations with Ca2+ were likely to indicate a primarily geogenic source of the solute. Elements with moderate to strong positive correlations with Ca2+ included SO42−, Ni2+, Rb+, Mo6+, and Ba2+. Inorganic N species, emanating primarily from an agricultural source, had negative relationships with Ca2+. Concentration-discharge relationships revealed chemostatic to weak dilution of analytes positively correlated with Ca2+, suggesting a geogenic source of these elements that was weakly diluted with increasing discharge. Cu2+ showed a positive concentration-discharge slope, suggesting an anthropogenic input, but a defined source was not identified. Dams on the Scioto River that form drinking water reservoirs dampened chemical correlations and concentration-discharge relationships. Both agricultural and urban sources influence the river's chemistry based on hydroclimatic conditions and land use. Higher frequency sampling of the Scioto River during high flows allowed for a more nuanced understanding of sources of solutes to the river compared to previous work. Continued water quality monitoring is imperative to maintain and protect water quality resources, especially as urban populations are predicted to grow significantly in the next century.