earticle

영어

To understand microbial metabolic functionality, it is important to focus on intracellular reaction kinetics. This requires the development of methodologies to measure intracellular reactant concentrations under in vivo conditions. The analysis of intracellular metabolites, nucleotides,
and cofactors has been attempted with a wide range of techniques, e.g., enzymatic assays, HPLC, GC-MS, LC-MS, and NMR. Although there are some limitations in using ESI LC-MS detection, it has a number of advantages compared to the other methods. Since glycolytic intermediates are especially difficult to analyze due to their polarity, structural similarity,
and noncharacteristic UV absorption. MS detection has not only the advantage of specific detection, but also that has m/z-specific separation. Here we propose a convenient and highly selective and sensitive LCMS/MS method using tributylamine as volatile ion pair reagent. The
method allows the combined separation and quantification of 9 metabolites from the most interesting biological classes in central carbon metabolism. This method can be successfully applied to identification and quantification of these intracellular metabolites in various cell
extract samples. In this study determination of intracellular metabolites in E. coli was given as an example.