Liquid chromatographic methods for the determination of endogenous nucleotides and nucleotide analogs used in cancer therapy: A review

Cohen, Sabine; Jordheim, Lars Petter; Megherbi, Mehdi; Dumontet, Charles; Guitton, Jérôme

doi:10.1016/j.jchromb.2010.05.016

Cited by 53 publications

(53 citation statements)

References 121 publications

(519 reference statements)

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“…Therefore, many chromatographic methods based on anion exchange or ion-pairing (IP) mechanisms have been developed over the last years [2][3][4][5]. In IP chromatography, stationary phases are usually C18 or porous graphitic carbon (PGC).…”

Section: Introductionmentioning

confidence: 99%

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: Introductionmentioning

confidence: 99%

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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“…In addition to being central building blocks for DNA and RNA [1], they also act as co-substrates in enzymatic reactions, play crucial roles in activating metabolites, and serve as signal mediators [2]. Furthermore, several intermediates of the purine biosynthesis pathway exhibit additional characteristics-ranging from flavor-enhancing qualities (inosine monophosphate (IMP) and guanosine monophosphate (GMP) [3,4]) to drug-assisted therapy (e.g., inosine and purine analogues [5,6]). These properties make them attractive targets for the pharmaceutical and biotechnological industries.…”

Section: Introductionmentioning

confidence: 99%

“…The latter has been used for many years with a broad application range in nucleotide analysis [2,5,11,[16][17][18][19]. The addition of cationic reagents such as alkylamines leads to formation of adducts between negatively charged nucleotides and positively charged ion pair reagent, increasing the retention factors for these compounds and thus improving retention.…”

Section: Introductionmentioning

confidence: 99%

Quantitation of intracellular purine intermediates in different Corynebacteria using electrospray LC-MS/MS

et al. 2012

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Intermediates of the purine biosynthesis pathway play key roles in cellular metabolism including nucleic acid synthesis and signal mediation. In addition, they are also of major interest to the biotechnological industry as several intermediates either possess flavor-enhancing characteristics or are applied in medical therapy. In this study, we have developed an analytical method for quantitation of 12 intermediates from the purine biosynthesis pathway including important nucleotides and their corresponding nucleosides and nucleobases. The approach comprised a single-step acidic extraction/quenching procedure, followed by quantitative electrospray LC-MS/MS analysis. The assay was validated in terms of accuracy, precision, reproducibility, and applicability for complex biological matrices. The method was subsequently applied for determination of free intracellular pool sizes of purine biosynthetic pathway intermediates in the two Gram-positive bacteria Corynebacterium glutamicum and Corynebacterium ammoniagenes. Importantly, no ion pair reagents were applied in this approach as usually required for liquid chromatography analysis of large classes of diverse metabolites.

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“…16,19 The extraction was generally performed at a low temperature; however, there were no effects on the recovery rate using a precooled solvent to -30 C (data not shown). Based on our investigations, the addition of ACN:water (70:30) extraction was found to be the best extraction technique.…”

Section: Resultsmentioning

confidence: 95%

“…16 These biochemicals have now been measured in a variety of systems, including cultured cells, [17][18][19] isolated tissues, 15 food substances, [12][13][14][20][21][22][23][24][25] fungi, 26 and biological samples. 11 Previously, we also reported the use of the HPLC method for determining the purine content of food, 14 beer, and beer-like alcoholic beverages, 12 whereas LC-MS was suitable for the simultaneous analysis of 23 species of purine bases, pyrimidines, nucleosides, and nucleotides.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Intra- and Extracellular Levels of Purine Bases, Nucleosides, and Nucleotides in HepG2 Cells by High-performance Liquid Chromatography

2015

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To evaluate cellular uptake and purine transport, we developed a high-performance liquid chromatography method for intra-and extracellular purine quantification. Our aim was to develop an effective method for simultaneously quantifying the substrate and metabolites with high sensitivity. C18 columns from different manufacturers were tested for simultaneous quantification of 22 different purine bases, nucleosides, and nucleotides. We used a YMC-Triart C18 column. The analysis conditions, including extraction solutions for the cells and cell culture medium, were optimized to achieve good quantification. Linearity, accuracy, determination limits, and recovery were assessed and showed good performance. The developed HPLC method was successfully applied to the qualitative analysis of 22 different intra-and extracellular purines, demonstrating that it is useful for studying the overall pattern of purine metabolism. This method could also be useful for evaluating metabolic dynamics of purines under a variety of stimulatory conditions of culture cells.

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Liquid chromatographic methods for the determination of endogenous nucleotides and nucleotide analogs used in cancer therapy: A review

Cited by 53 publications

References 121 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

Quantitation of intracellular purine intermediates in different Corynebacteria using electrospray LC-MS/MS

Analysis of Intra- and Extracellular Levels of Purine Bases, Nucleosides, and Nucleotides in HepG2 Cells by High-performance Liquid Chromatography

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