Liquid chromatographic method for the determination of uridine in human serum

Zilly, Michael; Langmann, Peter; Winzer, Ralf; Benesic, Andreas; Schirmer, Diana; Walker, Ulrich A.; Klinker, Hartwig

doi:10.1016/j.jchromb.2004.01.018

Cited by 4 publications

(4 citation statements)

References 9 publications

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“…The success of a quantitative LC/MS/MS method for determination of decitabine is largely dependent on the sample isolation and cleanup. The Oasis MCX SPE cartridge (Waters Corp.) has been used to extract basic compounds and was selected for extraction of decitabine from plasma because of its selectivity 45. Recently, an Oasis MCX SPE column was used to extract 5‐azacytidine from human plasma and showed acceptable recovery and accuracy for its quantification 46.…”

Section: Resultsmentioning

confidence: 99%

“…The Oasis MCX SPE cartridge (Waters Corp.) has been used to extract basic compounds and was selected for extraction of decitabine from plasma because of its selectivity. 45 Recently, an Oasis MCX SPE column was used to extract 5-azacytidine from human plasma and showed acceptable recovery and accuracy for its quantification. 46 We have successfully used this extraction column for decitabine.…”

Section: Sample Extraction and Optimization Of Hplc Conditionsmentioning

confidence: 99%

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Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

Liu

Marcucci

Byrd

et al. 2006

Rapid Comm Mass Spectrometry

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Aberrant DNA methylation patterns resulting in gene transcriptional repression are observed in numerous cancers. Decitabine, a DNA methyltransferase inhibitor, is being clinically evaluated in patients with hematologic malignancies and solid tumors. Decitabine is rather unstable and decomposes to 1-beta-D-2'-deoxyribofuranosyl-3-guanylurea under basic conditions and several additional unknown products under neutral conditions. This has greatly limited application of pharmacokinetic assays to clinical development of decitabine. In this paper, a high-performance liquid chromatography/ultraviolet multi-stage mass spectrometry (HPLC-UV-MSn) study of the decomposition of decitabine in water and human plasma revealed that these previously unknown products are isomers of the intermediates formyl-1-beta-D-2'-deoxyribofuranosyl-3-guanylurea and 1-beta-D-2'-deoxyribofuranosyl-3-guanylurea. A HPLC tandem mass spectrometry (MS/MS) method for the determination of decitabine concentrations in human and rat plasma has been developed. This method was based on a specific fragmentation pathway of the molecular ion of decitabine at m/z 229 to generate a unique fragment ion at m/z 113 under collision-induced dissociation. Separation of decitabine and the stable internal standard dihydro-5-aza-cytidine from the endogenous interfering substance in plasma extract was carried out on a C18 Aquasil column under an isocratic elution with a mobile phase consisting of 5% water/acetonitrile and 10 mM ammonium formate. The detection of decitabine was via selected reaction monitoring (SRM, 229 > 113), and its ionization was enhanced by post-column addition of acetonitrile. Effects of sample preparation and handling parameters on the stability of decitabine were also evaluated in human plasma at various temperatures. The accuracy and precision of this assay showed a coefficient of variation of <15% over the range of 0.5-25 ng for rat plasma and 0.1-25 ng for human plasma injected on-column. Pharmacokinetics of decitabine in rats following intravenous doses of 1.0 and 5.0 mg/kg were characterized. In the rat, plasma concentration-time profiles were found to follow a biexponential decline and the pharmacokinetics was dose-independent. Application of this decitabine pharmacokinetic assay to human studies is therefore justified and ongoing.

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Section: Resultsmentioning

confidence: 99%

Section: Sample Extraction and Optimization Of Hplc Conditionsmentioning

confidence: 99%

Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

Liu

Marcucci

Byrd

et al. 2006

Rapid Comm Mass Spectrometry

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“…RNA catabolism yields UMP and CMP, which can be converted into the corresponding NTPs via the successive action of CMPK1 and NDPK. With a plasma concentration of ~5 μM, uridine is the dominant circulatory nucleoside in mammals [10]; the plasma concentrations of all other pyrimidine nucleosides are at least an order of magnitude lower [11], and are therefore insufficient to support cellular demands of the corresponding nucleotides via direct salvage. Uridine/cytidine kinase (UCK) converts transported pyrimidine nucleosides into the corresponding NMPs, which can be further phosphorylated and modified as discussed above.…”

Section: Pyrimidine Nucleotide Biosynthesis Through De Novo and Salvamentioning

confidence: 99%

Human pyrimidine nucleotide biosynthesis as a target for antiviral chemotherapy

Ökesli

Khosla

Bassik

2017

Current Opinion in Biotechnology

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The development of broad-spectrum, host-acting antiviral therapies remains an important but elusive goal in anti-infective drug discovery. To replicate efficiently, viruses not only depend on their hosts for an adequate supply of pyrimidine nucleotides, but also up-regulate pyrimidine nucleotide biosynthesis in infected cells. In this review, we outline our understanding of mammalian de novo and salvage metabolic pathways for pyrimidine nucleotide biosynthesis. The available spectrum of experimental and FDA-approved drugs that modulate individual steps in these metabolic pathways is also summarized. The logic of a host-acting combination antiviral therapy comprised of inhibitors of dihydroorotate dehydrogenase and uridine/cytidine kinase is discussed.

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“…Anion gap was calculated from the electrolyte values using the formula ([Na + ] + [K + ]) -( [Cl -] + [HCO 3 -]), all units expressed in mmol/l. Serum uridine concentrations were measured using high performance liquid chromatography, as described in [30]. CD4…”

Section: Analytical Proceduresmentioning

confidence: 99%

Uridine Supplementation for the treatment of Antiretroviral Therapy-Associated Lipoatrophy: A Randomized, Double-Blind, Placebo-Controlled Trial

et al. 2007

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Background Highly active antiretroviral therapy (HAART) is associated with loss of subcutaneous fat (lipoatrophy) presumably due to mitochondrial toxicity of nucleoside reverse transcriptase inhibitors. In vitro, uridine abrogates thymidine analogue-induced toxicity in adipocytes. Methods A total of 20 patients with HAART-associated lipoatrophy were randomized to receive either a dietary uridine supplement (36 g three times a day for 10 consecutive days/month) or placebo, for 3 months. Body composition was measured using dual energy X-ray absorptiometry, magnetic resonance imaging and proton spectroscopy. Data are mean ± standard error of mean. Results The mean increases in limb fat (880 ±140 versus 230 ±270 g; P<0.05), intra-abdominal fat (210 ±80 versus -80 ±70 cm3; P<0.05) and total body fat (1,920 ±240 versus 240 ±520 g; P<0.01) were significantly greater in the uridine than in the placebo group. Within the uridine group, the changes from baseline to 3 months were statistically significant in total limb fat ( P<0.001), intra-abdominal fat ( P<0.05) and total body fat ( P<0.001). The proportion of limb fat to total fat increased from 18% to 25% ( P<0.05) in the uridine group. Liver fat content and lean body mass remained unchanged in both groups. High-density lipoprotein-cholesterol concentrations decreased in the uridine and increased in the placebo group, whereas fasting serum insulin concentrations did not change. Uridine supplementation was well tolerated and the virological effect of HAART was not affected. Conclusion Uridine supplementation significantly and predominantly increased subcutaneous fat mass in lipoatrophic HIV-infected patients during unchanged HAART.

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Liquid chromatographic method for the determination of uridine in human serum

Cited by 4 publications

References 9 publications

Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

Human pyrimidine nucleotide biosynthesis as a target for antiviral chemotherapy

Uridine Supplementation for the treatment of Antiretroviral Therapy-Associated Lipoatrophy: A Randomized, Double-Blind, Placebo-Controlled Trial

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