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Comparative changes in plasma, urine and sweat during endurance exercise using NMR-based metabolomics

William James O'Connor


Objective: The purpose of this study was to compare the relative metabolic profile of human biofluids before, during, and after strenuous endurance exercise. 

Method: Urine, blood, and sweat samples were collected from eleven healthy endurance trained cyclists at the beginning of, during, and immediately after a two hour cycle ergometer ride at 65% VO2peak. The exercised-induced metabolic changes in the sampled biofluids were followed using nuclear magnetic resonance (NMR) spectra recorded on a 700 MHz NMR spectrometer. Within the biofluids, group separation was accomplished based on relative changes in the distribution of metabolites based on their concentrations using a combination of principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Specific metabolites in the NMR spectra could be identified from their characteristic peak positions.

Results: Urine profiles showed significant pre- vs post-exercise differences based on relative metabolite concentration changes. The significant contributors to this difference were creatinine and acetoacetate, which increased following exercise, and glycine, citrate and alanine which decreased.  The differences seen in blood plasma were almost exclusively due to the substantial changes in glucose and lactate levels.  The most novel of the biofluids, sweat, interestingly showed pre-, mid-, and post- exercise differences due to significant increases in lactate and to a lesser extent, changes in pyruvate and glycerol.  In several instances plasma and sweat showed contrasting metabolite concentration changes across the exercise regimen.

Conclusion: The metabolic profile (metabolome) of plasma, sweat, and urine all show significant, yet differing responses to strenuous endurance exercise.


Biofluid, blood plasma, urine, NMR spectroscopy, metabolome

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AHLBORG, G., FELIG, P., HAGENFEL.L, HENDLER, R. & WAHREN, J. 1974. Substrate Turnover during Prolonged Exercise in Man - Splanchnic and Leg Metabolism of Glucose, Free Fatty-Acids, and Amino-Acids. Journal of Clinical Investigation, 53, 1080-1090. Available: ://A1974S546000015.

BUNTING, H., WISLOCKI, G. B. & DEMPSEY, E. W. 1948. The chemical histology of human eccrine and apocrine sweat glands. The Anatomical Record, 100, 61-77. Available: DOI 10.1002/ar.1091000106.

BUONO, M. J. 1999. Sweat ethanol concentrations are highly correlated with co-existing blood values in humans. Experimental Physiology, 84, 401-404. Available: ://000079657300013.

ESSÉN, B., HAGENFELDT, L. & KAIJSER, L. 1977. Utilization of blood-borne and intramuscular substrates during continuous and intermittent exercise in man. The Journal of Physiology, 265, 489-506. Available: http://jp.physoc.org/content/265/2/489.abstract.

FORTNEY, S. M., NADEL, E. R., WENGER, C. B. & BOVE, J. R. 1981. Effect of blood volume on sweating rate and body fluids in exercising humans. Journal of Applied Physiology, 51, 1594-1600. Available: http://jap.physiology.org/content/51/6/1594.abstract.

GORDON, R. S., THOMPSON, R. H., MUENZER, J. & THRASHER, D. 1971. Sweat Lactate in Man Is Derived from Blood Glucose. Journal of Applied Physiology, 31, 713-&. Available: ://A1971K827500014.

GREEN, J. M., PRITCHETT, R. C., CREWS, T. R., MCLESTER, J. R. & TUCKER, D. C. 2004. Sweat lactate response between males with high and low aerobic fitness. European Journal of Applied Physiology, 91, 1-6. Available: DOI DOI 10.1007/s00421-003-0968-2.

HARKER, M., COULSON, H., FAIRWEATHER, I., TAYLOR, D. & DAYKIN, C. A. 2006. Study of metabolite composition of eccrine sweat from healthy male and female human subjects by H-1 NMR spectroscopy. Metabolomics, 2, 105-112. Available: DOI DOI 10.1007/s11306-006-0024-4.

HAVEL, R. J., CARLSON, L. A., EKELUND, L.-G. & HOLMGREN, A. 1964. Turnover rate and oxidation of different free fatty acids in man during exercise. Journal of Applied Physiology, 19, 613-618. Available: http://jap.physiology.org/content/19/4/613.abstract.

HICKSON, R. C., RENNIE, M. J., CONLEE, R. K., WINDER, W. W. & HOLLOSZY, J. O. 1977. Effects of increased plasma fatty acids on glycogen utilization and endurance. Journal of Applied Physiology, 43, 829-833. Available: http://jap.physiology.org/content/43/5/829.abstract.

HOROWITZ, J. F. & KLEIN, S. 2000. Lipid metabolism during endurance exercise. The American Journal of Clinical Nutrition, 72, 558S-563S. Available: http://www.ajcn.org/content/72/2/558S.abstract.

HURLEY, B. F., NEMETH, P. M., MARTIN, W. H., HAGBERG, J. M., DALSKY, G. P. & HOLLOSZY, J. O. 1986. Muscle triglyceride utilization during exercise: effect of training. Journal of Applied Physiology, 60, 562-567. Available: http://jap.physiology.org/content/60/2/562.abstract.

JUEL, C. & HALESTRAP, A. P. 1999. Lactate transport in skeletal muscle — role and regulation of the monocarboxylate transporter. The Journal of Physiology, 517, 633-642. Available: DOI 10.1111/j.1469-7793.1999.0633s.x.

KUHL, J., MORITZ, T., WAGNER, H., STENLUND, H., LUNDGREN, K., BAVENHOLM, P., EFENDIC, S., NORSTEDT, G. & TOLLET-EGNELL, P. 2008. Metabolomics as a tool to evaluate exercise-induced improvements in insulin sensitivity. Metabolomics, 4, 273-282. Available: DOI DOI 10.1007/s11306-008-0118-2.

LIN, R.-Y., VERA, J. C., CHAGANTI, R. S. K. & GOLDE, D. W. 1998. Human Monocarboxylate Transporter 2 (MCT2) Is a High Affinity Pyruvate Transporter. Journal of Biological Chemistry, 273, 28959-28965. Available: DOI 10.1074/jbc.273.44.28959.

LINDON, J. C., NICHOLSON, J. K., HOLMES, E. & EVERETT, J. R. 2000. Metabonomics: Metabolic processes studied by NMR spectroscopy of biofluids. Concepts in Magnetic Resonance, 12, 289-320. Available: ://000088864500003.

LLOYD, D. P. C. 1959. Secretion and Reabsorption in Sweat Glands. Proceedings of the National Academy of Sciences of the United States of America, 45, 405-409. Available: ://A1959WJ53300023.

ORMSBEE, M. J., THYFAULT, J. P., JOHNSON, E. A., KRAUS, R. M., CHOI, M. D. & HICKNER, R. C. 2007. Fat metabolism and acute resistance exercise in trained men. Journal of Applied Physiology, 102, 1767-1772. Available: DOI 10.1152/japplphysiol.00704.2006.

PECHLIVANIS, A., KOSTIDIS, S., SARASLANIDIS, P., PETRIDOU, A., TSALIS, G., MOUGIOS, V., GIKA, H. G., MIKROS, E. & THEODORIDIS, G. A. 2010. H-1 NMR-Based Metabonomic Investigation of the Effect of Two Different Exercise Sessions on the Metabolic Fingerprint of Human Urine. Journal of Proteome Research, 9, 6405-6416. Available: DOI Doi 10.1021/Pr100684t.

REFSUM, H. E. & STRÖMME, S. B. 1974. Urea and Creatinine Production and Excretion in Urine during and after Prolonged Heavy Exercise. Scandinavian Journal of Clinical & Laboratory Investigation,

, 247-254. Available: DOI doi:10.1080/00365517409082493.

SAWKA, M. N., MONTAIN, S. J. & LATZKA, W. A. 2001. Hydration effects on thermoregulation and performance in the heat. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 128, 679-690. Available: DOI http://dx.doi.org/10.1016/S1095-6433(01)00274-4.

SHIBASAKI, M. & CRANDALL, C. G. 2010. Mechanisms and controllers of eccrine sweating in humans. Front Biosci (Schol Ed), 2, 685-96. Available: http://www.ncbi.nlm.nih.gov/pubmed/20036977.

TAYLOR, R. P., POLLIACK, A. A. & BADER, D. L. 1994. The analysis of metabolites in human sweat: analytical methods and potential application to investigation of pressure ischaemia of soft tissues. Ann Clin Biochem, 31 ( Pt 1), 18-24. Available: http://www.ncbi.nlm.nih.gov/pubmed/8154848.

WOLFE, S., CAGE, G., EPSTEIN, M., TICE, L., MILLER, H. & GORDON, R. S. 1970. Metabolic Studies of Isolated Human Eccrine Sweat Glands. Journal of Clinical Investigation, 49, 1880-&. Available: ://A1970H417300013.

YAN, B., A, J. Y., WANG, G. J., LU, H. L., HUANG, X. P., LIU, Y., ZHA, W. B., HAO, H. P., ZHANG, Y., LIU, L. S., GU, S. H., HUANG, Q., ZHENG, Y. T. & SUN, J. G. 2009. Metabolomic investigation into variation of endogenous metabolites in professional athletes subject to strength-endurance training. Journal of Applied Physiology, 106, 531-538. Available: DOI DOI 10.1152/japplphysiol.90816.2008.

ZEUTHEN, T. & KLAERKE, D. A. 1999. Transport of Water and Glycerol in Aquaporin 3 Is Gated by H+. Journal of Biological Chemistry, 274, 21631-21636. Available: DOI 10.1074/jbc.274.31.21631.

DOI: http://dx.doi.org/10.18103/mra.v2i3.255


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