Analysis of adenine nucleotides and nicotinamide coenzymes by CZE in combination with field enhanced stacking
| Authors | |
|---|---|
| Year of publication | 2008 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | 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. They have important functions in the coordination of metabolic processes and control. Determination of these metabolites is thus significant for metabolomic studies, because their pool analysis characterize the energetic state of the cell. Changes in environmental conditions can affect the total concentration of nucleotides and coezymes and influence the metabolism at several levels. As the importance of metabolome analysis was recognized, several metabolite analysis methods by means of GC-MS, LC-MS, NMR and FT-ICRMS have been developed. In addition to these well established methods capillary electrophoresis (CE) is gaining the position in this field. 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 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. |
| Related projects: |