Comprehensive analysis of the intracellular metabolism of antiretroviral nucleosides and nucleotides using liquid chromatography–tandem mass spectrometry and method improvement by using ultra performance liquid chromatography

Coulier, L; Gerritsen, H.W.; Kampen, Jeroen J. A. van; Reedijk, Mariska L.; Luider, Theo M.; Osterhaus, Albert D. M. E.; Gruters, Rob A.; Brull, L

doi:10.1016/j.jchromb.2011.07.045

Cited by 22 publications

(23 citation statements)

References 18 publications

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“…Since DEA Considering the presence of negative charges on phosphate groups, the use of negative mode appeared to be particularly relevant for nucleoside triphosphates. As reported in the literature, in negative mode, nucleoside triphosphates fragmented in a most abundant product ion at m/z 159, corresponding to the pyrophosphate fragment [HP 2 O 6 ] − [3,10,30]. In positive mode, nucleoside triphosphates formed a unique fragment corresponding to their base, except for UTP and thymidine triphosphate (TTP).…”

Section: Mass Spectrometrymentioning

confidence: 84%

Fully validated assay for the quantification of endogenous nucleoside mono- and triphosphates using online extraction coupled with liquid chromatography–tandem mass spectrometry

et al. 2014

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An analytical method coupling online solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed to quantify 16 endogenous nucleoside mono- and triphosphates in cellular samples. Separation was achieved on a porous graphitic carbon (PGC) column without ion-pairing agent in the mobile phase. Low levels of the ion-pairing agent diethylamine (DEA) added to the reconstitution solution were necessary to prevent peak tailing of nucleoside triphosphates. The mass spectrometer, a triple quadrupole with an electrospray ionisation source, was operated in positive mode. Two multiple reaction monitoring (MRM) segments were programmed, each an internal standard. Extraction and separation of nucleoside mono- and triphosphates were obtained within 20 min. The total duration of a single run was 37 min. Calibration curves, performed with labelled nucleotides added to the sample matrix, ranged from 0.29 to 18.8 pmol injected for deoxyribonucleotides and from 3.9 to 3,156 pmol for ribonucleotides. Accuracy did not deviate more than -14.6 and 10.2 % from nominal values for all compounds at all levels. CV results were all lower than 17.0 % for the LLOQ level and 14.6 % for the other levels. Quality control (QC) samples were also in agreement with acceptance criteria, except for the lower QC of GMP. Ion suppression, matrix effect, extraction recoveries and stability were assessed. After validation, the method was applied to the evaluation of the effects of gemcitabine and hydroxyurea on nucleotide pools in Messa cells.

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Section: Mass Spectrometrymentioning

confidence: 84%

Fully validated assay for the quantification of endogenous nucleoside mono- and triphosphates using online extraction coupled with liquid chromatography–tandem mass spectrometry

et al. 2014

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“…Special attention for chromatographic separation was only needed to resolve ADP from dGDP and ATP from dGTP, as each pair of compounds have the same precursor and product ion (Table 2) [21,23]. Sufficient chromatographic resolution was achieved with the present method to quantify each metabolite (Figure 1), therefore, reducing the risk of inaccuracy due to metabolite cross-talk and in-source fragmentation.…”

Section: Resultsmentioning

confidence: 99%

Comprehensive quantitative analysis of purines and pyrimidines in the human malaria parasite using ion-pairing ultra-performance liquid chromatography–mass spectrometry

Laourdakis

Merino

Neilson

et al. 2014

Journal of Chromatography B

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Targeted metabolite profiling has aided in the understanding of a variety of biological processes in the malaria parasite as well as in drug discovery. A fast and sensitive analytical method, based on ion-pairing reversed phase ultra-high performance liquid chromatography tandem mass spectrometry (IP-RP-UPLC-MS/MS), was optimized for the simultaneous analysis of intracellular levels of 35 purine and pyrimidine nucleobases, nucleosides, and nucleotides. This analytical method allows for chromatographic separation of highly polar metabolites using reverse phase chemistry within 15 minutes. The analytical performance of the method was evaluated and successfully applied to the quantification of purines and pyrimidines in Plasmodium falciparum and its host cell, the human erythrocyte. In addition, this method can be customized to include other related metabolites such as NADPH and NADP, among others.

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“…[23]. Human urine control (Medidrug Basis-line U) was purchased from LGC Standards (Teddington, UK).…”

Section: Methodsmentioning

confidence: 99%

“…Although UHPLC is relatively new technique it has already found many applications, e.g. nucleosides [23,24]. Utilization of UHPLC provided better resolution and moreover the time of analysis was reduced in great extent (2-5 times).…”

Section: Introductionmentioning

confidence: 99%

A new way to fast and high resolution determination of modified nucleosides

Studzińska

Buszewski

2012

Journal of Chromatography B

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Comprehensive analysis of the intracellular metabolism of antiretroviral nucleosides and nucleotides using liquid chromatography–tandem mass spectrometry and method improvement by using ultra performance liquid chromatography

Cited by 22 publications

References 18 publications

Fully validated assay for the quantification of endogenous nucleoside mono- and triphosphates using online extraction coupled with liquid chromatography–tandem mass spectrometry

Fully validated assay for the quantification of endogenous nucleoside mono- and triphosphates using online extraction coupled with liquid chromatography–tandem mass spectrometry

Comprehensive quantitative analysis of purines and pyrimidines in the human malaria parasite using ion-pairing ultra-performance liquid chromatography–mass spectrometry

A new way to fast and high resolution determination of modified nucleosides

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