One-quarter of anthropogenically produced nitrous oxide (N2O) comes from rivers and estuaries. Countries reporting N2O fluxes from aquatic surfaces under the United Nations Framework Convention on Climate Change typically estimate anthropogenic inorganic nitrogen loading and assume a fraction becomes N2O. However, several studies have not confirmed a linear relationship between dissolved nitrate (NO3-) and river N2O fluxes. We apply recursive partitioning analysis to examine the relationships between N2O flux and NO3-, dissolved oxygen (DO), temperature, land use and surficial geology in the Grand River, Canada, a seventh-order river in an agricultural catchment with substantial urban population. Results suggest that N2O flux is high when hypoxia exists. Temperature, not NO3-, was the primary correlate of N2O flux when hypoxia does not occur suggesting NO3- is not limiting N2O production and further increases in NO3- may not lead to comparable increases in N2O flux. This work indicates that a linear relationship between NO3- and N2O is unlikely to exist in most agricultural and urban impacted river systems. Most N2O is produced during hypoxia so quantifying the extent of hypoxia is a necessary first step to quantifying N2O fluxes in lotic systems. Predicted increases in riverine hypoxia via eutrophication and increased temperature due to climate change may drive nonlinear increases in N2O production.