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Analytical Chemistry
Real-Time Monitoring of Tricarboxylic Acid Metabolites in Exhaled Breath

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      Real-Time Monitoring of Tricarboxylic Acid Metabolites in Exhaled Breath
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      • Alberto Tejero Rioseras
        Alberto Tejero Rioseras
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
        SEADM, S.L., 28036 Madrid, Spain
        Department of Analytical Chemistry, University of Cordoba, 14005 Cordoba, Spain
      • Kapil Dev Singh
        Kapil Dev Singh
        University Children’s Hospital Basel, University of Basel, 4056 Basel, Switzerland
        Department of Biomedical Engineering, University of Basel, 4123 Allschwil, Switzerland
      • Nora Nowak
        Nora Nowak
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
        More byNora Nowak
      • Martin T. Gaugg
        Martin T. Gaugg
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
      • Tobias Bruderer
        Tobias Bruderer
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
      • Renato Zenobi
        Renato Zenobi
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
      • Pablo M.-L. Sinues*
        Pablo M.-L. Sinues
        Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland
        University Children’s Hospital Basel, University of Basel, 4056 Basel, Switzerland
        Department of Biomedical Engineering, University of Basel, 4123 Allschwil, Switzerland
        *E-mail:[email protected]

      Analytical Chemistry

      Cite this:Anal. Chem.2018, 90, 11, 6453–6460
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      PublishedMay 16, 2018
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      Copyright © 2018 American Chemical Society

      Abstract

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      The tricarboxylic acid (TCA) cycle is one of the most important metabolic pathway for cellular respiration in aerobic organisms. It provides and collects intermediates for many other interconnecting pathways and acts as a hub connecting metabolism of carbohydrates, fatty acids, and amino acids. Alteration in intracellular levels of its intermediates has been linked with a wide range of illnesses ranging from cancer to cellular necrosis or liver cirrhosis. Therefore, there exists an intrinsic interest in monitoring such metabolites. Our goal in this study was to evaluate whether, at least the most volatile metabolites of the TCA cycle, could be detected in breathin vivo and in real time. We used secondary electrospray ionization coupled with high-resolution mass spectrometry (SESI-HRMS) to conduct this targeted analysis. We enrolled six healthy individuals who provided full exhalations into the SESI-HRMS system at different times during 3 days. For the first time, we observed exhaled compounds that appertain to the TCA cycle: fumaric, succinic, malic, keto-glutaric, oxaloacetic, and aconitic acids. We found high intraindividual variability and a significant overall difference between morning and afternoon levels for malic acid, oxaloacetic acid, and aconitic acid, supporting previous studies suggesting circadian fluctuations of these metabolites in humans. This study provides first evidence that TCA cycle could conveniently be monitored in breath, opening new opportunities to studyin vivo this important metabolic pathway.

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      Copyright © 2018 American Chemical Society

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      Supporting Information

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      The Supporting Information is available free of charge on theACS Publications website at DOI:10.1021/acs.analchem.7b04600.

      • Figure S1, schematic description of the experimental setup; Figure S2, total ion current (TIC) as a function of time during the exhalation maneuvers; Figure S3, signal intensities vs time during two consecutives exhalations from the same subject; Figure S4, SESI-MS/MS fragmentation spectra of citric acid standard and breath; Figure S5, TCA cycle metabolites fumaric acid, succinic acid, and alpha-ketoglutaric acid showed no significant time-of-day dependence in exhaled breath abundance; Table S1, lock masses employed in negative ion mode to ensure high mass accuracy during the experiments; Table S2, list of chemicals used for running MS/MS experiments and collision energies used to compare the fragmentation pattern of the standards and the exhaled breath; Table S3, estimation of gas-phase concentrations in breath for healthy humans; Table S4, retention times and fragmentation spectra data fromFigure3 for EBC; Table S5, mean values, mean differences, corresponding 95% confidence interval, andp-value resulting from running an unpairedt test to compare morning vs afternoon levels of the TCA cycle metabolites identified in exhaled breath (PDF)

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      Cited By

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      This article is cited by 53 publications.

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      Analytical Chemistry

      Cite this:Anal. Chem.2018, 90, 11, 6453–6460
      Click to copy citationCitation copied!
      PublishedMay 16, 2018
      Copyright © 2018 American Chemical Society

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