Liquid chromatography–tandem mass spectrometry assays for intracellular deoxyribonucleotide triphosphate competitors of nucleoside antiretrovirals

• DOI: 10.1016/s1570-0232(03)00099-0
• Publication Date: 2003-05-13T01:09:12Z
• Authors: Gaëlle Henneré; François Becher; Alain Pruvost; Cécile Goujard; Jacques Grassi; Henri Benech
• Keywords: Nucleoside triphosphate; Deoxyribonucleotides; Deoxyribonucleotides; Ribonucleotide reductase; Deoxyribonucleosides; Ribonucleotide

This study was aimed to apply an LC-MS-MS method previously developed for intracellular nucleoside reverse transcriptase inhibitors-triphosphate (NRTI-TPs) to the determination of natural deoxyribonucleotides (dNTPs) in human peripheral blood mononuclear cells. The LC-MS-MS method was directly used in assay of dATP and dTTP. Interferences by ribonucleotides (rNTPs) prevented direct application to the two other analytes: dGTP and dCTP. A periodate oxidation procedure was therefore optimized to remove rNTPs from the cell medium in order to quantitate dCTP and dGTP. The determination of the intracellular ratio of NRTI-TP/dNTP in HIV-infected patients now involves use of the same chromatographic system for simultaneous assay of several analytes.