A Study of the Effects of Exercise on the Urinary Metabolome Using Normalisation to Individual Metabolic Output

Abstract

Aerobic exercise, in spite of its multi-organ benefit and potent effect on the metabolome, has yet to be investigated comprehensively via an untargeted metabolomics technology. We conducted an exploratory untargeted liquid chromatography mass spectrometry study to investigate the effects of a one-h aerobic exercise session in the urine of three physically active males. Individual urine samples were collected over a 37-h protocol (two pre-exercise and eight post-exercise). Raw data were subjected to a variety of normalization techniques, with the most effective measure dividing each metabolite by the sum response of that metabolite for each individual across the 37-h protocol expressed as a percentage. This allowed the metabolite responses to be plotted on a normalised scale. Our results highlight significant metabolites located in the following systems: purine pathway, tryptophan metabolism, carnitine metabolism, cortisol metabolism, androgen metabolism, amino acid oxidation, as well as metabolites from the gastrointestinal microbiome. Many of the significant changes observed in our pilot investigation mirror previous research studies, of various methodological designs, published within the last 15 years, although they have never been reported at the same time in a single study.

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

“…Similar results were also observed in the analysis of HX and XA in plasma where an increase in concentrations of both HX and XA were observed post-exercise, with a peak increase in the levels of HX post exercise of nearly nine times the resting level (Hellsten-Westing et al 1994). HX has been suggested as an indicator of the effectiveness of a training regime (Zielinski et al 2011), and as a potential biomarker to normalise for metabolomic changes due to exercise, although further research is required (Daskalaki et al 2015(Daskalaki et al , 2014).…”

Commentary: Metabolomics-Based Studies Assessing Exercise-Induced Alterations of the Human Metabolome: A Systematic Review

“…• Standardization of participant's preparation prior to collection of biological samples at rest and pre-exercise, since postprandial time, diet composition, and time after the previous training session are likely to affect the metabolome (Daskalaki et al, 2015;Shrestha et al, 2015;Karimpour et al, 2016;Giskeødegård et al, 2019). In this sense, we suggest the collection of biological samples at rest after 10-12 h overnight fasting or 90-120 min after a standardized meal previous to an exercise session, which is expected to present reasonable stabilization of postprandial metabolism (Shrestha et al, 2015;Karimpour et al, 2016;Giskeødegård et al, 2019;Li-Gao et al, 2019).…”

A method for the analysis of sugars in biological systems using reductive amination in combination with hydrophilic interaction chromatography and high resolution mass spectrometry

“…Urine samples were available from a previous study [20] and were collected in this study with permission from the University ethics committee. Post-mortem brain samples were also available from a previous study [21] and were provided by from the Sudden Death Bank collection held in the MRC Edinburgh Brain and Tissue Banks.…”