The N2O and N2 fluxes emitted from a temperate UK grassland soil after fertiliser application (equivalent to 25 and 75 kg N ha−1) were simultaneously measured, using a new automated soil incubation system, which replaces soil atmosphere (N2 dominated) with a He+O2 mixture. Dual isotope and isotopomer ratios of the emitted N2O were also determined. Total N2O and N2 fluxes were significantly lower (P < 0.001) in the control (0 kg N) than in the 25 and 75 kg N treatments. The total N2O flux was significantly higher (P < 0.001) in the 75 kg N than in the 25 kg N treatment. The general patterns of N2O and N2 fluxes were similar for both fertiliser treatments. The total gaseous N loss in the control treatment was nearly all N2, whereas in the fertiliser treatment more N2O than N2 was emitted from the soil. The ratio N2O/N2 fluxes as measured during the experiment suggested three phases in N2O production, in phase 1 nitrification > denitrification, in phase 2 denitrification > nitrification, and in phase 3 denitrification (and total denitrification) ≫ nitrification. Dual δ15N and δ18O isotope and isotopomer (δ15Nα and δ15Nβ) value ratios of emitted N2O also pointed towards an increasing dominance of the production of N2O by denitrification and total denitrification. The site preference value from the soil-emitted N2O was lower than the troposphere value. This confirmed that the enhanced troposphere N2O site preference could result from back injection of N2O from the stratosphere. The measurements of N2O/N2 flux ratio and the isotopic content of emitted N2O pointed, independently, to similar temporal trends in N2O production processes after fertiliser application to grassland soil. This confirmed that both measurements are suitable diagnostic tools to study the N2O production process in soils. Copyright © 2003 John Wiley & Sons, Ltd.