We have developed a new method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) for analysis of zidovudine-triphosphate and (deoxy)nucleotide-triphosphates, which ultimately can be used for nucleoside reverse transcriptase inhibitor (NRTI) treatment monitoring in HIV-1 infected children and adults. Four different matrices were compared for sensitivity and reproducibility of zidovudine-triphosphate detection and anthranilic acid mixed with nicotinic acid (AA/NA) was selected as most suitable matrix. Solutions of zidovudine-triphosphate, ATP, and dGTP were detected up to 0.5fmol per sample. Furthermore, intracellular zidovudine-triphosphate, ATP, and dGTP were detected in peripheral blood mononuclear cells (PBMCs). Zidovudine-triphosphate, ATP, and dGTP yield identical mass spectra, however MALDI-TOF post-source decay analysis can be used for discrimination between these compounds. We conclude that this method based on MALDI-TOF MS can be used for analysis of intracellular zidovudine-triphosphate and (deoxy)nucleotide-triphosphates in PBMCs.

Additional Metadata
Keywords Children, HIV-1/AIDS, Intracellular, MALDI, NRTI, Nucleotide, Triphosphate
Persistent URL dx.doi.org/10.1016/j.bbrc.2004.01.032, hdl.handle.net/1765/67239
Journal Biochemical and Biophysical Research Communications
Citation
van Kampen, J.J.A, Fraaij, P.L.A, Hira, V, van Rossum, A.M.C, Hartwig, N.G, de Groot, R, & Luider, T.M. (2004). A new method for analysis of AZT-triphosphate and nucleotide-triphosphates. Biochemical and Biophysical Research Communications, 315(1), 151–159. doi:10.1016/j.bbrc.2004.01.032