OBJECTIVE: To establish a reference interval for amino acid profiles for healthy red wolves (Canis rufus). METHODS: Heparinized plasma of 48 red wolves was collected between August 2023 and April 2024 and sent to the University of California-Davis Amino Acid Laboratory for analysis. Reference intervals were created using the published American Society for Veterinary Clinical Pathology reference interval guidelines. Data were analyzed via Gaussian data distribution, and parametric statistical methods were used to produce a 90% CI of reference limits. The means of the red wolf intervals created were compared to those of the domestic dog using a z test. RESULTS: Reference intervals were created for red wolves (n = 48). Upon completion of the z test, 11 of 21 amino acids were found to be statistically significantly different compared to those of the domestic dog. CONCLUSIONS: A reference interval was created for red wolves. The red wolf amino acid profiles are different than those of the domestic dog, with 52% (11/21) of the profiles being statistically different. CLINICAL RELEVANCE: Red wolf amino acid profiles should not be compared to those of the domestic dog reference intervals due to the significant difference between profiles.