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@proceedings{788057,
author = {Musilová, Jindra and Sedláček, Vojtěch and Pilátová, Kateřina and Kučera, Igor and Glatz, Zdeněk},
booktitle = {16th International Symposium on Capillary Electroseparation Techniques},
keywords = {metabolome analysis; extraction; nucleotide; coenzyme},
language = {eng},
title = {Development of quantitative sample preparation protocol for targeted metabolome analysis of adenine nucleotides and coenzymes by means of field enhanced sample stacking-capillary zone electrophoresis combination},
year = {2008}
}
TY - CONF
ID - 788057
AU - Musilová, Jindra - Sedláček, Vojtěch - Pilátová, Kateřina - Kučera, Igor - Glatz, Zdeněk
PY - 2008
TI - Development of quantitative sample preparation protocol for targeted metabolome analysis of adenine nucleotides and coenzymes by means of field enhanced sample stacking-capillary zone electrophoresis combination
KW - metabolome analysis
KW - extraction
KW - nucleotide
KW - coenzyme
N2 - Investigations in microbial metabolomics require reliable and reproducible analyses of the metabolites in the cell across a broad dynamic range of concentrations and from a range of different chemical functionalities. The development of robust a consistent experimental protocol for all steps in the procedure ranging from biomass cultivation, metabolism quenching, metabolites extraction to the quantitative analysis is required. Recently the selective and sensitive method for targeted metabolome analysis of adenine nucleotides and coenzymes based on field enhanced sample stacking-capillary zone electrophoresis combination has been developed. However such analytical method needs also appropriate methodology to isolate and to concentrate the target compounds from the biological matrix with minimal losses and this was the main aim of this work. Aqueous solutions containing organic solvents, such as methanol, ethanol, acetonitril and others, are widely used for extraction of intracellular metabolites from bacterial cells, because destabilize the cell wall and cell membrane proteins and lipids forming pores on the cell envelopes. In this consequence 20 - 100% ethanol, methanol, acetonitril, mixtures acetonitril/methanol/water (40:40:20), mixtures of acidic (0.1 M formic acid-containing) organic solvents and mixtures of basic (0.1 M NH4OH-containing) organic solvents were tested as an extraction media for adenine nucleotides (ATP, ADP, AMP) and coenzymes (NAD+, NADH, NADP+ and NADPH) from bacterium Paracoccus denitrificans and quantitative yields of metabolites were measured by capillary zone electrophoresis with UV detection in combination with field enhanced sample stacking and compared each other.
ER -
MUSILOVÁ, Jindra, Vojtěch SEDLÁČEK, Kateřina PILÁTOVÁ, Igor KUČERA a Zdeněk GLATZ. Development of quantitative sample preparation protocol for targeted metabolome analysis of adenine nucleotides and coenzymes by means of field enhanced sample stacking-capillary zone electrophoresis combination. Online. In \textit{16th International Symposium on Capillary Electroseparation Techniques}. 2008, [citováno 2024-04-23]
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