Analysis of serum parameters provides information about body condition, nutritional state, and health status of individuals/species, and has broad application in ecological research and veterinary diagnosis. This study establishes baseline values for serum chemistries of the Olrog's gull (Larus atlanticus). Glucose, urea, uric acid, total protein, globulin, albumin, cholesterol, triglyceride, calcium, and phosphorus concentrations were determined, as was the activity of the following enzymes: alkaline phosphatase, creatine phosphokinase, aspartate aminotransferase, and alanine aminotransferase. Thirty nonbreeding gulls (juvenile and subadult individuals) were captured and studied in Mar Chiquita Reserve (Buenos Aires, Argentina) during the wintering periods 2016 (n = 17) and 2017 (n = 13). In general terms, most values for the parameters reported were in line with those previously described for other seabirds. The year had a significant effect on several of the biochemical parameters evaluated, and the sex had a significant effect on the alkaline phosphatase and calcium. This study has defined the serum biochemical reference signatures for free-ranging Olrog's gulls during the nonbreeding period, and contributes to the knowledge of the overall health status of this threatened and endemic species.