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An Integrated Workflow for Qualitative Flux Analysis by Accurate Mass LC/MS
Metabolomics measures the abundance of metabolites, however, it lacks dynamic information. Increases in metabolite abundance can result from either increased production or decreased consumption and significant changes in flux through a pathway may not result in altered metabolite abundance. Stable isotope label tracing (qualitative flux analysis) has tremendous potential to help address these situations, and allow a deeper understanding of biological systems. In qualitative flux analysis, a stable isotope tracer (typically containing 13C, 15N, or 2H) is introduced into the biological system, and results in changes in the isotopic pattern (isotopologue distribution) of downstream metabolites. The approach is particularly useful in cell culture models that are amenable to the introduction of stable isotope tracers. Analysis of the resulting labeling pattern and kinetics of tracer incorporation provides insights into enzyme function, pathway dependence, and the effect of changes in gene expression or protein function.