Sixty seven serum samples collected from 43 European brown bears (Ursus arctos) from Croatia were tested for ≤31 serum chemistry parameters. Results were grouped and compared by bears origin (method of capture), sex, age, mass, and season sampled. Greatest differences were found between captive and free-living bears, and minor differences were found when sex, age, mass, or season of sampling were compared. Creatine kinase was significantly different among three categories of bears with the highest mean value of 924 IU/l in snare captured free-living bears compared to 67.8 IU/l in captive ones. Results of these tests provide reference values for European brown bears.