The Minnesota River, 10 major tributaries, and 21 springs were sampled to determine the sources and transport of sediment, nutrients, and oxygen- demanding substances. The study was part of a four-year assessment of non-point source pollution in the Minnesota River Basin. Runoff from tributary watersheds was identified as the primary source of suspended sediment and nutrients in the Minnesota River mainstem. Suspended-sediment, phosphorus, and nitrate concentrations were elevated in all major tributaries during runoff, but tributaries in the south-central and eastern part of the basin produce the highest annual loading to the mainstem because of higher annual precipitation and runoff in that part of the basin. Particle-size analyses showed that most of the suspended sediment in transport consisted of silt- and clay-size material. Phosphorus enrichment was indicated throughout the mainstem by total phosphorus concentrations that ranged from 0.04 to 0.48 mg/L with a median value of 0.22 mg/L, and an interquartile range of 0.15 to 0.29 mg/L. Nitrate concentrations periodically exceeded drinking water standards in tributaries draining the south-central and eastern part of the basin. Oxygen demand was most elevated during periods of summer low flow. Correlations between levels of biochemical oxygen demand and levels of algal productivity suggest that algal biomass comprises much of the oxygen-demanding material in the mainstem. Transport of sediment, nutrients, and organic carbon within the mainstem was found to be conservative, with nearly all tributary inputs being transported downstream. Uptake and utilization of nitrate and orthophosphorus was indicated during low flow, but at normal and high flow, inputs of these constituents greatly exceeded biological utilization.