Abstract
This chapter presents a simple, sensitive, and direct reverse phase (RP) liquid chromatography-mass spectrometry (LC-MS) method for the on-line identification and characterization of Ribavirin (1-β-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide, RTCONH 2 ) and its metabolites that does not require isolation and derivatization of individual metabolites. RTCONH 2 is a broad-spectrum antiviral agent is highly effective for the treatment of chronic hepatitis C viral infection when administered in combination with Intron A or PEG-Intron. Ribavirin and its metabolites are present as dephosphorylated moieties in plasma and excreta. Compounds that are phosphorylated intracellularly as with the case of ribavirin and its metabolites are hydrolyzed upon leaving the tissue and/or red blood cells. The method reported in the chapter provides on-line characterization of metabolites in biological samples using full scan LC-MS and LC-MS/MS techniques in conjunction with radiometric detection, thus accounting for all radiochromatographic peaks. This approach of simultaneous detection by a radiometric detector and LC-MS allows for semi-quantitative assessment and detection of all metabolites in a single experiment.
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