Abstract
A simple and sensitive method is described for the determination of methylaltrexone, a quaternary narcotic antagonist, in discrete rat brain regions and serum. Separation and quantitation are performed by reversedphase high-performance liquid chromatography with coulometric electrochemical detection. Perchloric acid extracts of the brain tissue or serum were passed through a normal-phase solid-phase extraction CN column, and methylnaltreexone was subsequently eluted with a mixture of absolute ethanol and 25mM trifluoroacetic acid. Methylnaltrexone gave a linear response over the range of 12.5–100ng in 1.0ml for cerebellar homogenates and 25–200ng in 50μl for serum. The average betweenassay coefficients of variation for methylnaltrexone from 1.0ml of cerebellar homogenate and 50μl of serum were 5.2 and 3.6%, respectively, over the concentration range studied. The within-assay coefficients of variation at 12.5ng/ml cerebellar homogenate and 25ng/50μl for serum were 7.4 and 4.6%, respectively. Analytical recovery of methylanyltrexone, added to cerebellar homogenate and serum samples, were 85 and 89%, respectively. The method was applied to the analysis of methylnaltrexone in discrete rat brain regions and serum after intraperitoneal injection.
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