Liquid chromatography-tandem mass spectrometry assays for intracellular deoxyribonucleotide triphosphate competitors of nucleoside antiretrovirals.
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.
CEA, Service de Pharmacologie et d'Immunologie, DSV/DRM, CEA/Saclay, 91191 Gif-Sur-Yvette, Cedex, France.
Gaëlle
G
Henneré
François
F
Becher
Alain
A
Pruvost
Cécile
C
Goujard
Jacques
J
Grassi
Henri
H
Benech
D002853:Chromatography, Liquid / Q000379:methods*; D003854:Deoxyribonucleotides / Q000032:analysis*; D015658:HIV Infections / Q000188:drug therapy; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000032:analysis* / Q000627:therapeutic use
D003854:Deoxyribonucleotides; D018894:Reverse Transcriptase Inhibitors