Saliva is increasingly being targeted for metabolic studies due to its non-invasive collection methods. Tracing levels of certain metabolites within biofluids can provide indications for a myriad of physiolog-ical conditions. This study was performed on a panel of eight analytes found in saliva that have shownassociations with physiological conditions of human performance, such as stress, inflammation, and circa-dian rhythm. This dual polarity liquid chromatography tandem mass spectrometric (LC-MS/MS) method was developed to accommodate a diverse group of analytes including steroids, alkaloids, and neurotrans-mitters. Samples collected during field exercises from soldiers were compared to those of civilians andbaseline levels of each of these compounds was determined in saliva. Although most analytes showed no significant differences between the two populations, relative cortisol levels were higher for soldiersthan for civilians. This developed dual polarity LC-MS/MS method can be applied to very diverse groupsof salivary analytes simultaneously.