Analgesia is an important part of veterinary medicine, but until recently there have been limited studies on analgesic drugs in avian species. Tramadol represents an orally administered opioid drug that has shown analgesic potential in numerous species, including mammals, birds, and reptiles. The objective of this study was to determine the pharmacokinetic parameters of tramadol and its primary metabolite, O-desmethyltramadol (M1), after oral administration of tramadol hydrochloride (HCl) in African penguins (Spheniscus demersus). A dose of 10 mg/kg of tramadol HCl was administered orally to 15 birds, and blood was collected at various time points from 0 to 36 hr. Tramadol and M1 concentrations were determined and were consistent with therapeutic concentrations in humans through 12 hr in 9/15 birds for tramadol and 36 hr in 14/15 birds for M1. Based on these findings and a comparison with other avian studies, an oral dose of 10 mg/kg of tramadol once daily appears to be a promising analgesic option for African penguins.