Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans
| Authors | |
|---|---|
| Year of publication | 2009 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Separation Science |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | Adenine Nucleotides and Coenzymes; Capillary zone electrophoresis; Field enhanced sample stacking; Metabolomics; Paracoccus denitrificans |
| Description | 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. 100 mM sodium carbonate/hydrogencarbonate buffer (pH 9.6) with the addition of beta-cyclodextrin at a concentration of 10 mM was found to be an effective background electrolyte for their separation within 20 minutes. Besides this, special attention was paid to the development of the procedure for the extraction of specific metabolites from the bacterium Paracoccus 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 on-line 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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