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Nontargeted Identification of Tracer Incorporation in High-Resolution Mass Spectrometry

  • Friederike Hoffmann
    Friederike Hoffmann
    Charité-Universitätsmedizin Berlin, Medical Department of Hematology, Oncology, and Tumor Immunology and Molekulares Krebsforschungszentrum (MKFZ), Augustenburger Platz 1, 13353 Berlin, Germany
  • Carsten Jaeger
    Carsten Jaeger
    Charité-Universitätsmedizin Berlin, Medical Department of Hematology, Oncology, and Tumor Immunology and Molekulares Krebsforschungszentrum (MKFZ), Augustenburger Platz 1, 13353 Berlin, Germany
    Berlin Institute of Health (BIH), Anna-Louisa-Karsch 2, 10178 Berlin, Germany
  • Animesh Bhattacharya
    Animesh Bhattacharya
    Charité-Universitätsmedizin Berlin, Medical Department of Hematology, Oncology, and Tumor Immunology and Molekulares Krebsforschungszentrum (MKFZ), Augustenburger Platz 1, 13353 Berlin, Germany
  • Clemens A. Schmitt
    Clemens A. Schmitt
    Charité-Universitätsmedizin Berlin, Medical Department of Hematology, Oncology, and Tumor Immunology and Molekulares Krebsforschungszentrum (MKFZ), Augustenburger Platz 1, 13353 Berlin, Germany
    Berlin Institute of Health (BIH), Anna-Louisa-Karsch 2, 10178 Berlin, Germany
    Max-Delbrück-Center for Molecular Medicine (MDC), Robert-Rössle-Straße 10, 13125 Berlin, Germany
  • , and 
  • Jan Lisec*
    Jan Lisec
    Federal Institute for Materials Research and Testing (BAM), Division 1.7 Analytical Chemistry, Richard-Willstätter-Straße 11, 12489 Berlin, Germany
    *E-mail: [email protected]. Fax: +49 (30) 8104-75891.
    More by Jan Lisec
Cite this: Anal. Chem. 2018, 90, 12, 7253–7260
Publication Date (Web):May 25, 2018
https://doi.org/10.1021/acs.analchem.8b00356
Copyright © 2018 American Chemical Society
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Supporting Info (8)»

Abstract

Abstract Image

“Fluxomics” refers to the systematic analysis of metabolic fluxes in a biological system and may uncover novel dynamic properties of metabolism that remain undetected in conventional metabolomic approaches. In labeling experiments, tracer molecules are used to track changes in the isotopologue distribution of metabolites, which allows one to estimate fluxes in the metabolic network. Because unidentified compounds cannot be mapped on pathways, they are often neglected in labeling experiments. However, using recent developments in de novo annotation may allow to harvest the information present in these compounds if they can be identified. Here, we present a novel tool (HiResTEC) to detect tracer incorporation in high-resolution mass spectrometry data sets. The software automatically extracts a comprehensive, nonredundant list of all compounds showing more than 1% tracer incorporation in a nontargeted fashion. We explain and show in an example data set how mass precision and other filter heuristics, calculated on the raw data, can efficiently be used to reduce redundancy and noninformative signals by 95%. Ultimately, this allows to quickly investigate any labeling experiment for a complete set of labeled compounds (here 149) with acceptable false positive rates. We further re-evaluate a published data set from liquid chromatography-electrospray ionization (LC-ESI) to demonstrate broad applicability of our tool and emphasize importance of quality control (QC) tests. HiResTEC is provided as a package in the open source software framework R and is freely available on CRAN.

Supporting Information

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The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.analchem.8b00356.

  • SI Table 1, evaluated candidate list of example data including annotation of likely False Positives due to inspection of QC plots from SI File 1 (XLSX)

  • SI Table 2, evaluated candidate list of reprocessed LC-MS data including annotation of likely false positives due to inspection of QC plots from SI File 3 (XLSX)

  • SI File 1, all QC plots for accepted candidates of ExampleData (PDF)

  • SI File 2, 100 randomly selected QC plots for rejected candidates of ExampleData (PDF)

  • SI File 3, all QC plots for accepted candidates of reanalyzed LC-MS data (geoRge) (PDF)

  • SI File 4, all QC plots for rejected candidates of reanalyzed LC-MS data (geoRge) (PDF)

  • SI File 5, all QC plots for accepted candidates of reanalyzed LC-MS data (de novo) (PDF)

  • SI Text, detailed information on QC filters and function parameters, GC-APCI processing and the mass drift phenomenon, and comparison of currently available tools for tracer incorporation detection (PDF)

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Cited By


This article is cited by 2 publications.

  1. Manohar C. Dange, Vivek Mishra, Bratati Mukherjee, Damini Jaiswal, Murtaza S. Merchant, Charulata B. Prasannan, Pramod P. Wangikar. Evaluation of freely available software tools for untargeted quantification of 13C isotopic enrichment in cellular metabolome from HR-LC/MS data. Metabolic Engineering Communications 2020, 10 , e00120. https://doi.org/10.1016/j.mec.2019.e00120
  2. Elizabeth M. Llufrio, Kevin Cho, Gary J. Patti. Systems-level analysis of isotopic labeling in untargeted metabolomic data by X13CMS. Nature Protocols 2019, 14 (7) , 1970-1990. https://doi.org/10.1038/s41596-019-0167-1

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