Development of capillary electrophoretic method for nucleotide and coenzyme analysis in bacterial and stem cells
| Authors | |
|---|---|
| Year of publication | 2012 |
| Type | Article in Proceedings |
| Conference | CHEMICA 50S |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | capillary electrophoresis, nucleotides, coenzymes |
| Description | Metabolite profiling is one of the major approaches for metabolomics studies. It involves the identification and quantitation of a group of metabolites common to a specific pathway or chemical class. Nucleotides and coenzymes represent central energetic metabolites in the complex metabolic network. As a consequence, the quantitative determination of such important compounds is fundamental in many areas of biochemical research. The concentrations of the nucleotides and coenzymes in living cells are in the micromolar concentration range therefore highly sensitive separation techniques are required for their determination. A combination of capillary zone electrophoresis (CZE) with probably the most extensively used on-line preconcentration technique – field enhanced sample stacking – allows their analysis in different cell types. Appropriate conditions for selective separation of 17 energetically important metabolites (purine, pyrimidine nucleotides, adenine coenzymes and Acetyl CoA) were found. The most dominant parameter for the analysis was pH of the background electrolyte. The optimized methodology was applied on the cell-extract of bacterium Paracoccus denitrificans and stem cell extract. The described method is suitable for determination of large group of energetically important metabolites and its utilisation depends on character of real cell samples. |
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