title: A new method for analysis of AZT-triphosphate and nucleotide-triphosphates.
abstract: 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.
authors.affiliation: Department of Paediatrics, Sophia Children's Hospital, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Jeroen J A
authors.initials: JJ
authors.lastname: van Kampen
authors.forename: Pieter L A
authors.initials: PL
authors.lastname: Fraaij
authors.forename: Vishal
authors.initials: V
authors.lastname: Hira
authors.forename: Annemarie M C
authors.initials: AM
authors.lastname: van Rossum
authors.forename: Nico G
authors.initials: NG
authors.lastname: Hartwig
authors.forename: Ronald
authors.initials: R
authors.lastname: de Groot
authors.forename: Theo M
authors.initials: TM
authors.lastname: Luider
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D019380:Anti-HIV Agents / Q000032:analysis* / Q000737:chemistry; D054306:Dideoxynucleotides; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000032:analysis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013942:Thymine Nucleotides / Q000032:analysis* / Q000737:chemistry; D015215:Zidovudine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000737:chemistry
chemical_list: D019380:Anti-HIV Agents; D054306:Dideoxynucleotides; D013942:Thymine Nucleotides; D015215:Zidovudine; C052873:zidovudine triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate