The giant anteater (Myrmecophaga tridactyla) is classified as a vulnerable species on Brazil’s list of species at risk of extinction mainly due to deforestation and forest fires. This has contributed to a considerable increase in detailed clinical case records of the treatment of wild species at veterinary institutions. However, the paucity of serum biochemical profiles of healthy giant anteaters has made it difficult to evaluate these animals, preventing diagnosis, treatment and prognosis. The objective of this work was to collect data about the biochemical profile of healthy giant anteaters from the Brazilian Cerrado raised in captivity, in order to better understand the physiological characteristics inherent to this species. Eighteen analytes from 12 healthy giant anteaters were measured. The following means and standard deviations were found in the biochemical analyses: albumin 3.29±0.33g/dL, ALT 15.49±7.98 IU/L, amylase 1037.92±149.04 IU/L, AST 21, 12±7.50 IU/L, total cholesterol 62.79±20.08mg/dL, HDL cholesterol 14.73±4.98mg/dL, LDL cholesterol 26.60±11.05mg/dL, VLDL cholesterol 2.14±1.06mg/dL, CK 111.61±70.16 IU/L, creatinine 1.05±0.37mg/dL, iron 194.64±81.17μg/dL, GGT 65.18±54.57 IU/L, glucose 103.71±29.63mg/dL, globulins 2.76±0.36g/dL, lipase 28.80±5.11 IU/L,TSP 6.05±0.56g/dL, triglycerides 10.71±5.29mg/dL, and urea 53.46±18.28mg/dL. The values found in this study can be used as references for the laboratory evaluation of giant anteaters living in conditions similar to those of this study. This is one of the first reports of biochemical examinations on giant anteaters of the Cerrado biome.