Methadone: radioimmunoassay and pharmacokinetics in the rat

A radioimmunoassay for the quantitation of methadone in biofluids is described. The antiserum was prepared by using an albumin conjugate of N-methyl-N-(1-methyl-3,3-diphenyl-4-oxohexyl)aminoethanol succinate. By employing tritium-labeled dl-methadone as the radioligand and a sample volume of 0.05 ml, the method has a lower limit of sensitivity of 3 ng/ml. The ability of the antiserum to detect methadone was not influenced by the presence of the metabolites of methadone or members of the methadol series. Morphine, codeine, levorphanol, meperidine, l, alpha-acetylmethadol and d-propoxyphene do not cross-react. After the i.v. administration of 0.90 or 1.5 mg/kg of methadone to male rats, plasma methadone levels decline biexponentially and elimination is independent of dose. The mean volume of distribution is 7.58 +/- 0.87 liters/kg; the mean elimination T 1/2 is 88.6 +/- 1.9 min and the plasma clearance is 59.3 +/- 1.4 ml/min/kg. These results demonstrate that methadone is eliminated by the rat much more rapidly than previously suspected.