BackgroundOral levothyroxine (l‐T4) supplementation is commonly used to treat hypothyroid dogs.ObjectivesInvestigate the plasma profile and pharmacokinetics of total thyroxine (tT4) after PO administration of a l‐T4 solution and its clinical efficacy in hypothyroid dogs.AnimalsTen dogs with naturally occurring hypothyroidism.MethodsAfter hypothyroidism diagnosis and supplementation with l‐T4 solution PO q24h at 20 μg/kg BW for minimum 4 weeks, the plasma profile and pharmacokinetics of tT4 were determined over 34 hours and the clinical condition of the dogs was evaluated.ResultsBefore dosing for pharmacokinetic evaluation, mean tT4 concentration was 23 ± 9 nmol/L. l‐T4 was absorbed rapidly (t max, 5 hours), reaching a mean maximal tT4 concentration of 56 ± 11 nmol/L. The apparent terminal half‐life was 11.8 hours. Clinical signs of hypothyroidism improved or resolved in all dogs after 4 weeks of treatment. The dosage of 20 μg/kg PO q24h was judged appropriate in 5 dogs, and 4 dogs required slight increases (9–16%). Twice daily treatment, with a 30% increase in dosage, was necessary for 1 dog.Conclusions and Clinical ImportanceThe pharmacokinetics of l‐T4 in hypothyroid dogs was similar to that reported in healthy euthyroid dogs. Clinical and hormonal responses to l‐T4 solution were rapid in all dogs. The starting dosage of 20 μg/kg PO q24h was suitable for maintenance supplementation in 50% of the dogs, minor dosage modification was required in 4 other dogs, and treatment q12h was required in 1 dog.