A new method for analysis of AZT-triphosphate and nucleotide-triphosphates.
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.
van Kampen
Jeroen J A
JJ
Department of Paediatrics, Sophia Children's Hospital, Erasmus MC, Rotterdam, The Netherlands.
Fraaij
Pieter L A
PL
Hira
Vishal
V
van Rossum
Annemarie M C
AM
Hartwig
Nico G
NG
de Groot
Ronald
R
Luider
Theo M
TM
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
D019380:Anti-HIV Agents; D054306:Dideoxynucleotides; D013942:Thymine Nucleotides; D015215:Zidovudine; C052873:zidovudine triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate