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The determination of salivary oxypurines before and after exercise by combined liquid chromatography-field asymmetric waveform ion mobility spectrometry-time-of-flight mass spectrometry

  • Original Research
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International Journal for Ion Mobility Spectrometry

Abstract

A method combining field asymmetric waveform ion mobility spectrometry with liquid chromatography-mass spectrometry (LC-FAIMS-MS) has been developed for the analysis of the oxypurine compounds hypoxanthine (HX) and xanthine (XA) in saliva. Separation of the oxypurines from interfering matrix components was investigated using FAIMS-MS. The selected FAIMS parameters were then applied to the rapid LC-FAIMS-MS analysis of HX and XA using a short chromatographic separation method (7 min). A comparison of the LC-MS method with and without FAIMS applied, resulted in improved discrimination from saliva matrix interferences and improved chromatographic peak integration for both HX and XA using a FAIMS separation. A quantitative evaluation of the LC-FAIMS-MS method was performed giving limits of detection of 2.0 ng mL−1 for HX and 1.8 ng mL−1 for XA, and limits of quantification of 6.6 ng mL−1 for HX and 6.0 ng mL−1 for XA. The developed LC-FAIMS-MS method was applied to the targeted analysis of the oxypurine metabolites in saliva collected from healthy male athletes (n = 11) before and after exercise designed to induce oxidative stress; post-exercise collection time-points included immediately after exercise, one hour and twenty-four hours’ post-exercise. The salivary concentrations of both HX and XA were lower after physical exercise, compared to the pre-exercise (rest) concentrations and returned to approximately pre-exercise levels after twenty-four hours. The method reported has the potential for monitoring the salivary oxypurines, HX and XA, as biomarkers of oxidative stress and in other clinical applications.

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Acknowledgements

The authors thank Owlstone Ltd. and Loughborough University for financial support, and Owlstone Ltd. and Agilent Technologies for the provision of instrumentation.

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Authors and Affiliations

  1. Centre for Analytical Science, Department of Chemistry, Loughborough University, Epinal Way, Loughborough, Leicestershire, LE11 3TU, UK

    Kayleigh L. Arthur, Matthew A. Turner, James C. Reynolds & Colin S. Creaser

  2. Translational Chemical Biology research group, SSEHS, Loughborough University, Epinal Way, Loughborough, Leicestershire, LE11 3TU, UK

    Lynsey S. Wilson & Martin R. Lindley

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  1. Kayleigh L. Arthur
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  2. Lynsey S. Wilson
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  3. Matthew A. Turner
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  4. Martin R. Lindley
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  5. James C. Reynolds
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  6. Colin S. Creaser
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Correspondence to James C. Reynolds or Colin S. Creaser.

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Arthur, K.L., Wilson, L.S., Turner, M.A. et al. The determination of salivary oxypurines before and after exercise by combined liquid chromatography-field asymmetric waveform ion mobility spectrometry-time-of-flight mass spectrometry. Int. J. Ion Mobil. Spec. 21, 87–95 (2018). https://doi.org/10.1007/s12127-018-0232-4

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  • DOI: https://doi.org/10.1007/s12127-018-0232-4

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