(464e) Non-Targeted Metabolomics for the Elucidation of Metabolic Pathways

Systems level tools for the quantitative analysis of metabolic networks are required to engineer metabolism for biomedical and industrial applications. While current metabolomics techniques enable high-throughput quantification of metabolites, these methods provide minimal information on the rates and connectivity of metabolic pathways. Here we present a new method, Non-targeted Tracer Fate Detection (NTFD), that expands upon the concept of metabolomics to solve the above problems. Through the combined use of stable isotope tracers and chromatography coupled to mass spectrometry, our computational analysis enables the quantitative detection of all measurable metabolites derived from a specific labeled compound. Without a priori knowledge of a reaction network or compound library, NTFD provides information about relative flux magnitudes into each metabolite pool by determining the mass isotopomer distribution for all labeled compounds. We demonstrate the utility of this method by identifying fluxes and pathways specifically upregulated in tumor cells. This system adds a new dimension to the metabolomics tool box and provides a framework for global the analysis of metabolic fluxes.