A highly sensitive high-performance liquid chromatography–mass spectrometry method for quantification of fludarabine triphosphate in leukemic cells

Kalhorn, Thomas F.; Ren, Aaron G.; Slattery, John T.; Wang, Joanne

doi:10.1016/j.jchromb.2005.03.034

Cited by 17 publications

(22 citation statements)

References 15 publications

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“…Moreover, these active metabolites are determined at the target site. For this reason, nucleotide analogs active against hepatitis, acquired immunodeficiency syndrome (AIDS), and leukemia have been measured (both in vitro and in vivo) in hepatocytes, peripheral blood mononuclear cells (PBMCs), and leukemic blasts, respectively (Claire, 2000;Ray et al, 2004;Kalhorn et al, 2005). Matrices associated with toxicity have also been analyzed, such as erythrocytes in ribavirin therapy (Yeh et al, 2007).…”

Section: A Matrixmentioning

confidence: 99%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

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Nucleoside analogs are widely used in anti-cancer, anti-(retro)viral, and immunosuppressive therapy. Nucleosides are prodrugs that require intracellular activation to mono-, di-, and finally triphosphates. Monitoring of these intracellular nucleotides is important to understand their pharmacology. The relatively involatile salts and ion-pairing agents traditionally used for the separation of these ionic analytes limit the applicability of mass spectrometry (MS) for detection. Both indirect and direct methods have been developed to circumvent this apparent incompatibility. Indirect methods consist of de-phosphorylation of the nucleotides into nucleosides before the actual analysis. Various direct approaches have been developed, ranging from the use of relatively volatile or very low levels of regular ion-pairing agents, hydrophilic interaction chromatography (HILIC), weak anion-exchange, or porous graphitic carbon columns to capillary electrophoresis and matrix-assisted light desorption--time of flight (MALDI-TOF) MS. In this review we present an overview of the publications describing the quantitative analysis of therapeutic intracellular nucleotide analogs using MS. The focus is on the different approaches for their direct analysis. We conclude that despite the technical hurdles, several useful MS-compatible chromatographic approaches have been developed, enabling the use of the excellent selectivity and sensitivity of MS for the quantitative analysis of intracellular nucleotides.

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Section: A Matrixmentioning

confidence: 99%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

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“…The triphosphate metabolite of fludarabine, F-ara-ATP, was extracted from the biological matrix (cells, PBMC, lymphocytes) by protein precipitation with PCA and neutralization with KOH [101,102] or KHCO 3 [103,104]. It was shown that these compounds were stable during at least 12 h in the neutralized supernatant [103].…”

Section: Sample Pre-treatmentmentioning

confidence: 99%

“…It was shown that these compounds were stable during at least 12 h in the neutralized supernatant [103].…”

Section: Sample Pre-treatmentmentioning

confidence: 99%

“…Like other nucleosidic analog triphosphates or endogenous nucleoside triphosphates, the assays for the determination of Fara-phosphates were based on IP-HPLC [103] or IE-HPLC [101,102] or RP-HPLC [104]. The literature showed that UV detection lacked of sensitivity and specificity and required a delicate and reproducible chromatographic separation corresponding to long run times [101,102] since the synthetic nucleosides and their phosphorylated metabolites had similar UV properties to those of endogenous nucleosides and nucleotides.…”

Section: High Performance Liquid Chromatography and Detectionmentioning

confidence: 99%

“…According to these studies, the wavelength used for fludarabine triphosphate was 261 nm [101] or 246 nm [102] and for fludarabine monophosphate 272 nm [105]. Mass spectrometry, operating in the negative ionisation mode, appeared to be the detection mode of choice [103,104].…”

Section: High Performance Liquid Chromatography and Detectionmentioning

confidence: 99%

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

Cohen

Jordheim

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et al. 2010

Journal of Chromatography B

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Integrin repertoire on myogenic cells changes during the course of primary myogenesis in the mouse

Cachaço

Pereira

Pardal

et al. 2005

Developmental Dynamics

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Here, we determined ␣ subunit expression patterns at embryonic day (E) 11.5-E14.5. Differentiated myotomal myocytes express all ␣ subunits studied. As the muscle masses form both in trunk (E12.5) and limbs (E11.5-E12.5), laminin receptors ␣6␤1 and ␣7␤1 are undetectable, and an assembled laminin matrix is absent. Instead ␣1␤1, ␣4␤1, ␣5␤1, and an ␣v-containing integrin are expressed and unassembled laminin and fibronectin are abundant around myogenic cells. At E13.5-E14.5, ␣6␤1 and ␣7␤1 are expressed, and a laminin matrix forms around individual myotubes. Thus, myogenic cells change their integrin expression pattern during the course of primary myogenesis in the mouse, suggesting different roles for fibronectin-and laminin-containing matrices in this process.

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A highly sensitive high-performance liquid chromatography–mass spectrometry method for quantification of fludarabine triphosphate in leukemic cells

Cited by 17 publications

References 15 publications

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

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

Integrin repertoire on myogenic cells changes during the course of primary myogenesis in the mouse

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