title: A three-minute method for high-throughput quantitative metabolomics and quantitative tracing experiments of central carbon and nitrogen pathways.
abstract: BACKGROUND
  The implementation of mass spectrometry (MS)-based metabolomics is advancing many areas of biomedical research. The time associated with traditional chromatographic methods for resolving metabolites prior to mass analysis has limited the potential to perform large-scale, highly powered metabolomics studies and clinical applications.
  
  
  METHODS
  Here we describe a three-minute method for the rapid profiling of central metabolic pathways through UHPLC/MS, tracing experiments in vitro and in vivo, and targeted quantification of compounds of interest using spiked in heavy labeled standards.
  
  
  RESULTS
  This method has shown to be linear, reproducible, selective, sensitive, and robust for the semi-targeted analysis of central carbon and nitrogen metabolism. Isotopically labeled internal standards are used for absolute quantitation of steady-state metabolite levels and de novo synthesized metabolites in tracing studies. We further propose exploratory applications to biofluids, cell and tissue extracts derived from relevant biomedical/clinical samples.
  
  
  CONCLUSIONS
  While limited to the analysis of central carbon and nitrogen metabolism, this method enables the analysis of hundreds of samples per day derived from diverse biological matrices. This approach makes it possible to analyze samples from large patient populations for translational research, personalized medicine, and clinical metabolomics applications. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, 12801 East 17th Ave, Aurora, CO, 80045, USA.
authors.forename: Travis
authors.initials: T
authors.lastname: Nemkov
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, 12801 East 17th Ave, Aurora, CO, 80045, USA.
authors.forename: Kirk C
authors.initials: KC
authors.lastname: Hansen
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, 12801 East 17th Ave, Aurora, CO, 80045, USA.
authors.forename: Angelo
authors.identifier: http://orcid.org/0000-0002-2258-6490
authors.initials: A
authors.lastname: D'Alessandro
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D001826:Body Fluids / Q000737:chemistry; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D002244:Carbon / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D013058:Mass Spectrometry / Q000379:methods; D055432:Metabolomics / Q000379:methods*; D009584:Nitrogen / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D002244:Carbon; D009584:Nitrogen