A Specific Radioimmunoassay for the Determination of Morphine-6-Glucuronide in Human Plasma

Abstract

A specific antiserum for morphine-6-glucuronide (M6G) has been raised in a rabbit in response to immunization with a novel hapten: Protein conjugate (N-aminobutylnormorphine-6-glucuronide-thyroglobulin). Cross-reactivity with morphine and structurally related compounds was found to be negligible as expected from the nature of this immunogen. Using this antiserum, a simple, rapid and robust radioimmunoassay (RIA) has been developed for determination of M6G in samples of human plasma. The assay has a sensitivity of 0·05 ng/mL using 100 μL sample volumes and affords complete recovery of M6G over the range 2–200 ng/mL. The presence of morphine or morphine-3-glucuronide at concentrations up to 100 times the levels of M6G did not result in any measurable interference. Close agreement was obtained between M6G results obtained using the RIA and a specific high-performance liquid chromatography assay. This RIA offers an attractive alternative to existing methods for the determination of M6G in human plasma and will facilitate further metabolic and pharmacokinetic studies of morphine and M6G in the clinical setting.