River water has been suggested as a potential source of nitrous oxide (N2O), which is a greenhouse gas that is accumulating rapidly in the troposphere and which is a precursor to stratospheric NOx that depletes ozone. Previous studies on freshwater N2O sources have specifically examined estuaries where sedimentary N2O production might be important and a few points near anthropogenic nitrogen sources such as agricultural or municipal wastewater areas. Here we present the first observation of a temporal and horizontal distribution of N2O and its isotopomers between the midstream and estuary of an urban river. Surface water was supersaturated (100–6800%) with N2O at all stations during the study period. The average or maximum saturation value was greater than described in most previous reports. High N2O concentrations were observed near sewage plants and the unique signature of isotopomer ratios implied direct N2O addition from the plants. The isotopomer ratios also suggested N2O production/consumption at the sediment-water interface. Fluxes and isotopomer ratios of N2O, from the river to the atmosphere, estimated from our observations, indicated that the urban river is indeed a source of atmospheric N2O and that its production could be distinguished from other natural or anthropogenic sources using isotopomer ratios. Copyright © 2009 John Wiley & Sons, Ltd.