High performance liquid chromatography determination of dextromethorphan and its metabolites in urine using...

A high performance liquid chromatography assay coupled with fluorescence detection was developed for the determination of dextromethorphan and its metabolites in urine. The products and the internal standard, pholcodine, were separated on an Alltima C18, 5 microns column (250 x 4.6 mm), using a mobile phase containing sodium dodecyl sulphate (1 mM) in a mixture of acetonitrile-sodium dihydrogen phosphate (0.01 M) 40.5:59.5, v/v) (pH* = 2.5). A novel solid-phase extraction procedure with strong cation exchange, non end-capped, Isolute SCX cartridges allows good recovery of the products (mean 85% or more). For all analytes, the assay is sensitive (LOQ 25 ng ml-1, using 200 microliters urine), reproducible (RSD < 15%) and accurate (< 15% deviation of the nominal value) over the range evaluated. This method can be used to measure dextromethorphan and its metabolites to phenotype individuals as poor or extensive metabolizers of drugs metabolized by CYP2D6.