Abstract. The relationship between antecedent flow conditions and nitrate concentrations was explored at eight sites in the 2.9 million square kilometers (km2) Mississippi River basin, USA. Antecedent flow conditions were quantified as the ratio between the mean daily flow of the previous year and the mean daily flow from the period of record (Qratio), and the Qratio was statistically related to nitrate anomalies (the unexplained variability in nitrate concentration after filtering out season, long-term trend, and contemporaneous flow effects) at each site. Nitrate anomaly and Qratio were negatively related at three of the four major tributary sites and upstream in the Mississippi River, indicating that when mean daily streamflow during the previous year was lower than average, nitrate concentrations were higher than expected. The strength of these relationships increased when data were subdivided by contemporaneous flow conditions. Five of the eight sites had significant negative relationships (p ≤ 0.05) at high or moderately high contemporaneous flows, suggesting nitrate that accumulates in these basins during a drought is flushed during subsequent high flows. At half of the sites, when mean daily flow during the previous year was 50 percent lower than average, nitrate concentration can be from 9 to 27 percent higher than nitrate concentrations that follow a year with average mean daily flow. Conversely, nitrate concentration can be from 8 to 21 percent lower than expected when flow during the previous year was 50 percent higher than average. Previously documented for small, relatively homogenous basins, our results suggest that relationships between antecedent flows and nitrate concentrations are also observable at a regional scale. Relationships were not observed (using all contemporaneous flow data together) for basins larger than 1 million km2, suggesting that above this limit the overall size and diversity within these basins may necessitate the use of more complicated statistical approaches or that there may be no discernible basin-wide relationship with antecedent flow. The relationships between nitrate concentration and Qratio identified in this study serve as the basis for future studies that can better define specific hydrologic processes occurring during and after a drought (or high flow period) which influence nitrate concentration, such as the duration or magnitude of low flows, and the timing of low and high flows.