Metabolomic analysis of easily accessible biofluids has provided numerous biomarkers in urine and blood for biodosimetric purposes. In this pilot study we assessed saliva for its utility in biodosimetry using a mouse model. Mice were exposed to 0.5, 3 and 8 Gy total-body gamma irradiation and saliva was collected on day 1 and 7 postirradiation. Global metabolomic profiling was conducted through liquid chromatography mass spectrometry and metabolites were positively identified using tandem mass spectrometry. Multivariate data analysis revealed distinct metabolic profiles for all groups at day 1, whereas at day 7 the two lower dose profiles appeared to have minimal differences. Metabolites that were identified include amino acids and fatty acids, and intermediates of the nicotinate and nicotinamide metabolism. The specificity and sensitivity of the radiation signature, as expected, was higher for the 8 Gy dose at both time points, as determined through generation of receiver operating characteristic curves. To the best of our knowledge, this is the first metabolomics study in saliva of irradiated mice to demonstrate the utility of this biofluid as a potential matrix for identification of radiation and dose-specific biomarkers.