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The objective of this study was to establish the clinical pharmacokinetic profile of 4 different opioid drugs (buprenorphine, butorphanol, hydromorphone, and morphine) in the domestic ferret (Mustela putorius furo). Twenty-four, approximately 1-year-old, male neutered purpose-bred domestic ferrets were used for this study. The ferrets were divided into 4 groups of 6, with a different opioid drug used for each group. A preopioid venous blood sample was obtained via cranial vena cava venipuncture. Following the initial blood collection, a single injection of opioid (hydromorphone 0.1 mg/kg, buprenorphine 0.04 mg/kg, butorphanol 0.3 mg/kg, and morphine 1 mg/kg) was given to each ferret, dependent on assigned drug group, intramuscularly (buprenorphine) or subcutaneously (hydromorphone, butorphanol, and morphine). Intramuscular injections were administered in the semimembranosis and semitendinosis muscles, whereas the subcutaneous injections were delivered in the intrascapular subcutaneous space. A venous blood sample was obtained at 5, 15, 30, 60, 120, 240, 360, 480, and 720 minutes postinjection from the ferrets in the buprenorphine, butorphanol, and hydromorphone groups. Mass spectrometry and liquid chromatography was performed to obtain plasma concentrations of the administered drugs. The mean maximum concentration of buprenorphine was 6.96 ng/mL, butorphanol was 48.6 ng/mL, and hydromorphone was 17.3 ng/mL. Maximum concentrations were achieved at a mean of 9 minutes after administration for buprenorphine, 13.3 minutes for butorphanol, and 8.33 minutes for hydromorphone. The mean half-life of buprenorphine was 219.1 minutes, butorphanol was 91.1 minutes, and hydromorphone was 24.7 minutes. Owing to severe complications arising within the morphine group, including hypersalivation and vomiting, the morphine study was discontinued prior to blood sample collection. Intramuscular injections of buprenorphine and subcutaneous injections of butorphanol or hydromorphone appeared to be well tolerated by all ferrets. The pharmacokinetics of buprenorphine, butorphanol, and hydromorphone of a single equipotent dose of each drug have been established through this research investigation and may be useful for further studies.