Staphylococcus aureus is considered to be an osmo-tolerant bacterium (Shebuski et al., 2000; Vilhelmsson and Miller, 2002a, b), however, little is understood about the mechanisms conferring high salt tolerance by the bacteria. Our recent study investigated amino acid metabolites and proteomic responses of S. aureus when incubated in a minimal medium supplemented with 5% NaCl. The results indicated significant alterations in amino acid concentrations and protein regulations (Alreshidi et al., 2019). However, to date this experimental design has yet to be adjusted to investigate nucleotides, nucleosides, organic acids and sugar alcohol levels in response to the conditions of low temperature and/or high NaCl. Therefore, this study aims to characterize the metabolic changes (specifically in reference to carbohydrates; pyrimidine and purine bases; nucleosides and nucleotides) that occur as a result of low temperature (4°C) and elevated sodium chloride (5%) conditions. It is hypothesized that the exposure of S. aureus to conditions of low temperature and elevated NaCl induce significant alterations in its metabolic profiles.