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The microbial mediated process of denitrification is a major pathway for the removal of reactive nitrogen a pollutant in natural environments and waste treatment facilities. Denitrification potential, measured as denitrification enzyme activity (DEA), was quantified in novel short-term (4 h) anaerobic assays using a more sensitive and precise technique, membrane inlet mass spectrometry (MIMS) rather than the traditional headspace electron capture gas chromatography (GC-ECD) method. Using MIMS modifications made to the instrument and sample handling allowed for the simultaneous and direct measurement of reaction products nitrous oxide (N2O), a potent greenhouse gas, and the chemically unreactive dinitrogen (N2).