2007
Analysis of adenine nucleotides and nicotinamide coenzymes by CZE in combination with field enhanced stacking
MUSILOVÁ, Jindra, Igor KUČERA a Zdeněk GLATZZákladní údaje
Originální název
Analysis of adenine nucleotides and nicotinamide coenzymes by CZE in combination with field enhanced stacking
Název česky
Analýza adeninových nukleotidů a nikotinamidových koenzymů pomocí CZE
Autoři
Vydání
1. vyd. Brno, Book of Abstracts "CECE2007 4th International Interdisciplinary Meeting on Bioanalysis" od s. 44-44, 1 s. 2007
Nakladatel
Institute of Analytical Chemistry AS CR
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Česká republika
Utajení
není předmětem státního či obchodního tajemství
Organizační jednotka
Přírodovědecká fakulta
ISBN
978-80-254-0427-0
Klíčová slova anglicky
Metabolomics; Capillary Zone Electrophoresis; field enhanced stacking;
Příznaky
Mezinárodní význam
Změněno: 27. 6. 2008 10:32, Mgr. Jindra Smutná, Ph.D.
V originále
Being the intermediates of biochemical reactions, metabolites play a very important role in connecting the many different pathways that operate within living cell. Among them adenine nucleotides and nicotinamide coenzymes are central energetic metabolites in this complex metabolic network. Determination of these metabolites is thus important for metabolomic studies, because their pool analysis characterize the energetic state of the cell under a variety of physiological conditions during cell growth. A method for determination of adenine nucleotides (ATP, ADP, AMP) and nicotinamide coenzymes (NAD+, NADH, NADP+, NADPH) by CE was developed. As low concentrations of these metabolites are presented in the cellular extracts the on-line preconcentration technique field enhanced stacking was combined with capillary zone electrophoresis (CZE) to enhance the concentration sensitivities. The determination was performed in a 75 mm fused silica capillary using separation voltage 18 kV (positive polarity), temperature of capillary 20 oC and direct detection at 260 and 340 nm. 100 mM ammonium carbonate buffer (pH 9,6) was used as the background electrolyte, but addition of beta-cyclodextrin up to 5 mM concentration was required for better resolution of given analytes. Metabolites samples were dissolved in deionised water and injected into the capillary hydrodynamically with a pressure of 35 mbar for 20 s. Under these conditions, the detection limits were in the range of 300 to 400 nM at a signal-to-noise ratio (S/N) of 3. The optimized methodology was applied on the cell extract of Paracoccus denitrificans. The concentrations of given metabolites in the bacterial cells under different growth conditions were determined.
Česky
Byla vypracována metoda pro stanovení adeninových nukleotidů a nikotinamidových koenzymů pomocí CZE v kombinaci s technikou "field enhanced stacking". Metoda byla aplikována na buněčný extrakt bakterie Paracoccus denitrificans.
Návaznosti
| GA203/06/1179, projekt VaV |
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| LC06023, projekt VaV |
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| MSM0021622413, záměr |
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