Analysis of nucleotide pools in human lymphoma cells by capillary electrophoresis

Shao, Xiaowen; O’Neill, Kim L.; Zhao, Zhiwei; Anderson, Sean C.; Malik, Abdul; Lee, Milton

doi:10.1016/0021-9673(94)85144-1

Cited by 28 publications

(27 citation statements)

References 25 publications

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“…Due to the biochemical significance of such compounds, several papers have been published on their determination in different biological sources using CE. Shao et al [19] (5), NADPH (6) and ATP (7). Concentration of individual standard was 10 lM.…”

Section: Cze Methods Developmentmentioning

confidence: 99%

Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans

Musilová

Sedláček

Kučera

et al. 2009

J of Separation Science

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The main aim of this work was to demonstrate the applicability of capillary zone electrophoresis in combination with field enhanced sample stacking in targeted metabolome analyses of adenine nucleotides--AMP, ADP, ATP, coenzymes NAD(+), NADP(+) and their reduced forms in Paracoccus denitrificans. Sodium carbonate/hydrogencarbonate buffer (100 mM, pH 9.6) with the addition of beta-CD at a concentration of 10 mM was found to be an effective BGE for their separation within 20 min. Besides this, special attention was paid to the development of the procedure for the extraction of specific metabolites from the bacterium P. denitrificans. This procedure was not only optimised to achieve the highest metabolite yields but also to obtain a sample that was fully compatible with the online preconcetration strategy used. The developed methodology was finally applied in a study of the bacterium P. denitrificans at various stages of the active respiratory chain.

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Section: Cze Methods Developmentmentioning

confidence: 99%

Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans

Musilová

Sedláček

Kučera

et al. 2009

J of Separation Science

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“…Various chromatographic approaches have been proposed, including zwitterion and anion-exchange chromatography by HPLC (9 -12), a combination of techniques (13), or more recently CE (1,14,15). Ionpair chromatography on reversed phase was quite successful at separating NTP (1,16) or phosphorylated anticancer or antiviral nucleoside analogues (17)(18)(19)(20).…”

mentioning

confidence: 99%

Simultaneous Determination of Deoxyribonucleoside in the Presence of Ribonucleoside Triphosphates in Human Carcinoma Cells by High-Performance Liquid Chromatography

Décosterd

Cottin

Chen

et al. 1999

Analytical Biochemistry

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“…It has a high mass sensitivity in relation to the small (typically nanoliter) injection volume used. However, because of the short optical path length within the detection cell, the lowest detectable concentration in CE with UV absorption detection is in the 1-10-mM range [23,39,40]. This concentration sensitivity is lower than with HPLC.…”

Section: Resultsmentioning

confidence: 89%

A highly sensitive CE‐UV method with dynamic coating of silica‐fused capillaries for monitoring of nucleotide pyrophosphatase/phosphodiesterase reactions

et al. 2008

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A new highly sensitive capillary electrophoresis (CE) method applying dynamic coating and on-line stacking for the monitoring of nucleotide pyrophosphatases/phosphodiesterases (NPPs) and the screening of inhibitors was developed. NPP1 and NPP3 are membrane glycoproteins that catalyze the hydrolysis nucleotides, e.g. convert adenosine 5'-triphosphate to adenosine 5'-monophosphate (AMP) and pyrophosphate. Enzymatic reactions were performed and directly subjected to CE analysis. Since the enzymatic activity was low, standard methods were insufficient. The detection of nanomolar AMP and other nucleotides could be achieved by field-enhanced sample injection and the addition of polybrene to the running buffer. The polycationic polymer caused a dynamic coating of the silica-fused capillary, resulting in a reversed electroosmotic flow. The nucleotides migrated in the direction of the electroosmotic flow, whereas the positively charged polybrene molecules moved in the opposite direction, resulting in a narrow sample zone over a long injection time. Using this on-line sensitivity enhancement technique, a more than 70-fold enrichment was achieved for AMP (limit of detection, 46 nM) along with a short migration time (5 min) without compromising separation efficiency and peak shape. The optimized CE conditions were as follows: fused-silica capillary (30 cm effective lengthx75 mum), electrokinetic injection for 60 s, 50 mM phosphate buffer pH 6.5, 0.002% polybrene, constant current of -60 microA, UV detection at 210 nm, uridine 5'-monophosphate as the internal standard. The new method was used to study enzyme kinetics and inhibitors. It opens an easy way to determine the activities of slowly metabolizing enzymes such as NPPs, which are of considerable interest as novel drug targets.

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Analysis of nucleotide pools in human lymphoma cells by capillary electrophoresis

Cited by 28 publications

References 25 publications

Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans

Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans

Simultaneous Determination of Deoxyribonucleoside in the Presence of Ribonucleoside Triphosphates in Human Carcinoma Cells by High-Performance Liquid Chromatography

A highly sensitive CE‐UV method with dynamic coating of silica‐fused capillaries for monitoring of nucleotide pyrophosphatase/phosphodiesterase reactions

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