European Journal of Applied Physiology

, Volume 113, Issue 7, pp 1673–1684 | Cite as

Dietary nitrate supplementation improves team sport-specific intense intermittent exercise performance

  • Lee J. Wylie
  • Magni Mohr
  • Peter Krustrup
  • Sarah R. Jackman
  • Georgios Ermιdis
  • James Kelly
  • Matthew I. Black
  • Stephen J. Bailey
  • Anni Vanhatalo
  • Andrew M. JonesEmail author
Original Article


Recent studies have suggested that dietary inorganic nitrate (NO3 ) supplementation may improve muscle efficiency and endurance exercise tolerance but possible effects during team sport-specific intense intermittent exercise have not been examined. We hypothesized that NO3 supplementation would enhance high-intensity intermittent exercise performance. Fourteen male recreational team-sport players were assigned in a double-blind, randomized, crossover design to consume 490 mL of concentrated, nitrate-rich beetroot juice (BR) and nitrate-depleted placebo juice (PL) over ~30 h preceding the completion of a Yo–Yo intermittent recovery level 1 test (Yo–Yo IR1). Resting plasma nitrite concentration ([NO2 ]) was ~400 % greater in BR compared to PL. Plasma [NO2 ] declined by 20 % in PL (P < 0.05) and by 54 % in BR (P < 0.05) from pre-exercise to end-exercise. Performance in the Yo–Yo IR1 was 4.2 % greater (P < 0.05) with BR (1,704 ± 304 m) compared to PL (1,636 ± 288 m). Blood [lactate] was not different between BR and PL, but the mean blood [glucose] was lower (3.8 ± 0.8 vs. 4.2 ± 1.1 mM, P < 0.05) and the rise in plasma [K+] tended to be reduced in BR compared to PL (P = 0.08). These findings suggest that NO3 supplementation may promote NO production via the nitrate-nitrite-NO pathway and enhance Yo–Yo IR1 test performance, perhaps by facilitating greater muscle glucose uptake or by better maintaining muscle excitability. Dietary NO3 supplementation improves performance during intense intermittent exercise and may be a useful ergogenic aid for team sports players.


Repeated high-intensity exercise Nitric oxide Team sports Ergogenic aids 



The authors thank Jamie R Blackwell for advice and assistance with the plasma nitrate and nitrite analyses.


  1. Allen DG, Lamb GD, Westerblad H (2008) Skeletal muscle fatigue: cellular mechanisms. Physiol Rev 88:287–332PubMedCrossRefGoogle Scholar
  2. Allen JD, Stabler T, Kenjale A, Ham KL, Robbins JL, Duscha BD, Dobrosielski DA, Annex BH (2010) Plasma nitrite flux predicts exercise performance in peripheral arterial disease after 3 months of exercise training. Free Radic Biol Med 49:1138–1144PubMedCrossRefGoogle Scholar
  3. Atkins SJ (2006) Performance of the Yo–Yo intermittent recovery test by elite professional and semiprofessional rugby league players. J Strength Cond Res 20:222–225PubMedGoogle Scholar
  4. Bailey SJ, Winyard P, Vanhatalo A, Blackwell JR, DiMenna FJ, Wilkerson DP, Tarr J, Benjamin N, Jones AM (2009) Dietary nitrate supplementation reduces the O2 cost of low-intensity exercise and enhances tolerance to high-intensity exercise in humans. J Appl Physiol 107:1144–1155PubMedCrossRefGoogle Scholar
  5. Bailey SJ, Fulford J, Vanhatalo A, Winyard PG, Blackwell JR, DiMenna FJ, Wilkerson DP, Benjamin N, Jones AM (2010) Dietary nitrate supplementation enhances muscle contractile efficiency during knee-extensor exercise in humans. J Appl Physiol 109:135–148PubMedCrossRefGoogle Scholar
  6. Bailey SJ, Vanhatalo A, Winyard PG, Jones AM (2011) The nitrate-nitrite-nitric oxide pathway: its role in human exercise physiology. Eur J Sport Sci 12:309–320CrossRefGoogle Scholar
  7. Bangsbo J, Mohr M (2012) Fitness testing in football. Bangsbosport ISBN 978-87-994880-0-1, Copenhagen, DenmarkGoogle Scholar
  8. Bangsbo J, Krustrup P, González-Alonso J, Saltin B (2001) ATP production and efficiency of human skeletal muscle during intense exercise: effect of previous exercise. Am J Physiol Endocrinol Metab 280:E956–E964PubMedGoogle Scholar
  9. Bangsbo J, Iaia FM, Krustrup P (2007) Metabolic response and fatigue in soccer. Int J Sports Physiol Perform 2:111–127PubMedGoogle Scholar
  10. Bangsbo J, Iaia FM, Krustrup P (2008) The Yo–Yo intermittent recovery test: a useful tool for evaluation of physical performance in intermittent sports. Sports Med 38:37–51PubMedCrossRefGoogle Scholar
  11. Behnke BJ, McDonough P, Padilla DJ, Musch TI, Poole DC (2003) Oxygen exchange profile in rat muscles of contrasting fibre types. J Physiol 549:597–605PubMedCrossRefGoogle Scholar
  12. Bescós R, Ferrer-Roca V, Galilea PA, Roig A, Drobnic F, Sureda A, Martorell M, Cordova A, Tur JA, Pons A (2012) Sodium nitrate supplementation does not enhance performance of endurance athletes. Med Sci Sports Exerc 44:2400–2409Google Scholar
  13. Bond H, Morton L, Braakhuis AJ (2012) Dietary nitrate supplementation improves rowing performance in well-trained rowers. Int J Sport Nutr Exerc Metab 22:251–256PubMedGoogle Scholar
  14. Bradley PS, Mohr M, Bendiksen M, Randers MB, Flindt M, Barnes C, Hood P, Gomez A, Andersen JL, Di Mascio M, Bangsbo J, Krustrup P (2011) Sub-maximal and maximal Yo–Yo intermittent endurance test level 2: heart rate response, reproducibility and application to elite soccer. Eur J Appl Physiol 111:969–978PubMedCrossRefGoogle Scholar
  15. Burnley M, Jones AM (2007) Oxygen uptake kinetics as determinant of sports performance. Eur J Sport Sci 7:63–79CrossRefGoogle Scholar
  16. Cermak NM, Gibala MJ, van Loon LJ (2012a) Nitrate supplementation’s improvement of 10-km time-trial performance in trained cyclists. Int J Sport Nutr Exerc Metab 22:64–71PubMedGoogle Scholar
  17. Cermak NM, Res P, Stinkens R, Lundberg JO, Gibala MJ, van Loon L JC (2012b) No improvement in endurance performance following a single dose of beetroot juice. Int J Sport Nutr Exerc Metab 22:470–478Google Scholar
  18. Christensen PM, Nyberg M, Bangsbo J (2013) Influence of nitrate supplementation on VO(2) kinetics and endurance of elite cyclists. Scand J Med Sci Sports 23:e21–e31Google Scholar
  19. Colliander EB, Dudley GA, Tesch PA (1988) Skeletal muscle fiber type composition and performance during repeated bouts of maximal, concentric contractions. Eur J Appl Physiol Occup Physiol 58:81–86PubMedCrossRefGoogle Scholar
  20. Cosby K, Partovi KS, Crawford JH, Patel RP, Reiter CD, Martyr S, Yang BK, Waclawiw MA, Zalos G, Xu X, Huang KT, Shields H, Kim-Shapiro DB, Schechter AN, Cannon RO 3rd, Gladwin MT (2003) Nitrite reduction to nitric oxide by deoxyhemoglobin vasodilates the human circulation. Nat Med 9:1498–1505PubMedCrossRefGoogle Scholar
  21. Dreissigacker U, Wendt M, Wittke T, Tsikas D, Maassen N (2010) Positive correlation between plasma nitrite and performance during high-intensive exercise but not oxidative stress in healthy men. Nitric Oxide 23:128–135PubMedCrossRefGoogle Scholar
  22. Ferguson SK, Hirai DM, Copp SW, Holdsworth CT, Allen JD, Jones AM, Musch TI, Poole DC (2013) Impact of dietary nitrate supplementation via beetroot juice on exercising muscle vascular control in rats. J Physiol 591:547–557Google Scholar
  23. Gladwin MT, Shelhamer JH, Schechter AN, Pease-Fye ME, Waclawiw MA, Panza JA, Ognibene FP, Cannon RO 3rd (2000) Role of circulating nitrite and S-nitrosohemoglobin in the regulation of regional blood flow in humans. Proc Natl Acad Sci USA 97:11482–11487PubMedCrossRefGoogle Scholar
  24. Govoni M, Jansson EA, Weitzberg E, Lundberg JO (2008) The increase in plasma nitrite after a dietary nitrate load is markedly attenuated by an antibacterial mouthwash. Nitric Oxide 19:333–337PubMedCrossRefGoogle Scholar
  25. Greenhaff PL, Nevill ME, Soderlund K, Bodin K, Boobis LH, Williams C, Hultman E (1994) The metabolic responses of human type I and II muscle fibres during maximal treadmill sprinting. J Physiol 478:149–155PubMedGoogle Scholar
  26. Hernández A, Schiffer TA, Ivarsson N, Cheng AJ, Bruton JD, Lundberg JO, Weitzberg E, Westerblad H (2012) Dietary nitrate increases tetanic [Ca2+]i and contractile force in mouse fast-twitch muscle. J Physiol 590:3575–3583PubMedCrossRefGoogle Scholar
  27. Hogan MC, Richardson RS, Haseler LJ (1999) Human muscle performance and PCr hydrolysis with varied inspired oxygen fractions: a 31P-MRS study. J Appl Physiol 86:1367–1373PubMedGoogle Scholar
  28. Hord NG, Tang Y, Bryan NS (2009) Food sources of nitrates and nitrites: the physiologic context for potential health benefits. Am J Clin Nutr 90:1–10PubMedCrossRefGoogle Scholar
  29. Iaia FM, Thomassen M, Kolding H, Gunnarsson T, Wendell J, Rostgaard T, Nordsborg N, Krustrup P, Nybo L, Hellsten Y, Bangsbo J (2008) Reduced volume but increased training intensity elevates muscle Na+–K+ pump alpha1-subunit and NHE1 expression as well as short-term work capacity in humans. Am J Physiol Regul Integr Comp Physiol 294:R966–R974PubMedCrossRefGoogle Scholar
  30. Ingebrigtsen J, Bendiksen M, Randers MB, Castagna C, Krustrup P, Holtermann A (2012) Yo–Yo IR2 testing of elite and sub-elite soccer players: performance, heart rate response and correlations to other interval tests. J Sports Sci 30:1337–1345PubMedCrossRefGoogle Scholar
  31. Kapil V, Milsom AB, Okorie M, Maleki-Toyserkani S, Akram F, Rehman F, Arghandawi S, Pearl V, Benjamin N, Loukogeorgakis S, Macallister R, Hobbs AJ, Webb AJ, Ahluwalia A (2010) Inorganic nitrate supplementation lowers blood pressure in humans: role for nitrite-derived NO. Hypertension 56:274–281PubMedCrossRefGoogle Scholar
  32. Kenjale AA, Ham KL, Stabler T, Robbins JL, Johnson JL, Vanbruggen M, Privette G, Yim E, Kraus WE, Allen JD (2011) Dietary nitrate supplementation enhances exercise performance in peripheral arterial disease. J Appl Physiol 110:1582–1591PubMedCrossRefGoogle Scholar
  33. Krustrup P, Mohr M, Amstrup T, Rysgaard T, Johansen J, Steensberg A, Pedersen PK, Bangsbo J (2003) The Yo–Yo intermittent recovery test: physiological response, reliability, and validity. Med Sci Sports Exerc 35:697–705PubMedCrossRefGoogle Scholar
  34. Krustrup P, Söderlund K, Mohr M, Bangsbo J (2004) The slow component of oxygen uptake during intense, sub-maximal exercise in man is associated with additional fibre recruitment. Pflugers Arch 447:855–866PubMedCrossRefGoogle Scholar
  35. Krustrup P, Mohr M, Steensberg A, Bencke J, Kjaer M, Bangsbo J (2006) Muscle and blood metabolites during a soccer game: implications for sprint performance. Med Sci Sports Exerc 38:1165–1174PubMedCrossRefGoogle Scholar
  36. Krustrup P, Söderlund K, Relu MU, Ferguson RA, Bangsbo J (2009) Heterogeneous recruitment of quadriceps muscle portions and fibre types during moderate intensity knee-extensor exercise: effect of thigh occlusion. Scand J Med Sci Sports 19:576–584PubMedCrossRefGoogle Scholar
  37. Lansley KE, Winyard PG, Bailey SJ, Vanhatalo A, Wilkerson DP, Blackwell JR, Gilchrist M, Benjamin N, Jones AM (2011a) Acute dietary nitrate supplementation improves cycling time trial performance. Med Sci Sports Exerc 43:1125–1131PubMedCrossRefGoogle Scholar
  38. Lansley KE, Winyard PG, Fulford J, Vanhatalo A, Bailey SJ, Blackwell JR, DiMenna FJ, Gilchrist M, Benjamin N, Jones AM (2011b) Dietary nitrate supplementation reduces the O2 cost of walking and running: a placebo-controlled study. J Appl Physiol 110:591–600PubMedCrossRefGoogle Scholar
  39. Larsen FJ, Weitzberg E, Lundberg JO, Ekblom B (2007) Effects of dietary nitrate on oxygen cost during exercise. Acta Physiol (Oxf) 91:59–66CrossRefGoogle Scholar
  40. Larsen FJ, Weitzberg E, Lundberg JO, Ekblom B (2010) Dietary nitrate reduces maximal oxygen consumption while maintaining work performance in maximal exercise. Free Radic Biol Med 48:342–347PubMedCrossRefGoogle Scholar
  41. Larsen FJ, Schiffer TA, Borniquel S, Sahlin K, Ekblom B, Lundberg JO, Weitzberg E (2011) Dietary inorganic nitrate improves mitochondrial efficiency in humans. Cell Metab 13:149–159PubMedCrossRefGoogle Scholar
  42. Liu TH, Wu CL, Chiang CW, Lo YW, Tseng HF, Chang CK (2009) No effect of short-term arginine supplementation on nitric oxide production, metabolism and performance in intermittent exercise in athletes. J Nutr Biochem 20:462–468PubMedCrossRefGoogle Scholar
  43. Lundberg JO, Weitzberg E (2010) NO-synthase independent NO generation in mammals. Biochem Biophys Res Commu 396:39–45CrossRefGoogle Scholar
  44. Lundberg JO, Carlström M, Larsen FJ, Weitzberg E (2011) Roles of dietary inorganic nitrate in cardiovascular health and disease. Cardiovasc Res 89:525–532PubMedCrossRefGoogle Scholar
  45. Masschelein E, Van Thienen R, Wang X, Van Schepdael A, Thomis M, Hespel P (2012) Dietary nitrate improves muscle but not cerebral oxygenation status during exercise in hypoxia. J Appl Physiol 113:736–745PubMedCrossRefGoogle Scholar
  46. McDonough P, Behnke BJ, Padilla DJ, Musch TI, Poole DC (2005) Control of microvascular oxygen pressures in rat muscles comprised of different fibre types. J Physiol 563:903–913PubMedCrossRefGoogle Scholar
  47. McKenna MJ, Bangsbo J, Renaud JM (2008) Muscle K+, Na+, and Cl disturbances and Na+–K+ pump inactivation: implications for fatigue. J Appl Physiol 104:288–295PubMedCrossRefGoogle Scholar
  48. Merry TL, Lynch GS, McConell GK (2010) Downstream mechanisms of nitric oxide-mediated skeletal muscle glucose uptake during contraction. Am J Physiol Regul Integr Comp Physiol 299:R1656–R1665PubMedCrossRefGoogle Scholar
  49. Mohr M, Krustrup P, Bangsbo J (2003) Match performance of high standard soccer players with special reference to development of fatigue. J Sports Sci 21:519–528PubMedCrossRefGoogle Scholar
  50. Mohr M, Krustrup P, Nielsen JJ, Nybo L, Rasmussen MK, Juel C, Bangsbo J (2007) Effect of two different intense training regimens on skeletal muscle ion transport proteins and fatigue development. Am J Physiol Regul Integr Comp Physiol 292:R1594–R1602PubMedCrossRefGoogle Scholar
  51. Mohr M, Nordsborg N, Nielsen JJ, Pedersen LD, Fischer C, Krustrup P, Bangsbo J (2004) Potassium kinetics in human muscle interstitium during repeated intense exercise in relation to fatigue. Pflugers Arch 448:452–456PubMedCrossRefGoogle Scholar
  52. Mohr M, Nielsen JJ, Bangsbo J (2011) Caffeine intake improves intense intermittent exercise performance and reduces muscle interstitial potassium accumulation. J Appl Physiol 111:1372–1379PubMedCrossRefGoogle Scholar
  53. Murphy M, Eliot K, Heuertz RM, Weiss E (2012) Whole beetroot consumption acutely improves running performance. J Acad Nutr Diet 112:548–552PubMedCrossRefGoogle Scholar
  54. Nielsen OB, de Paoli FV (2007) Regulation of Na+–K+ homeostasis and excitability in contracting muscles: implications for fatigue. Appl Physiol Nutr Metab 32:974–984PubMedCrossRefGoogle Scholar
  55. Nielsen JJ, Kristensen M, Hellsten Y, Bangsbo J, Juel C (2003) Localization and function of ATP-sensitive potassium channels in human skeletal muscle. Am J Physiol Regul Integr Comp Physiol 284:R558–R563PubMedGoogle Scholar
  56. Nordsborg N, Mohr M, Pedersen LD, Nielsen JJ, Langberg H, Bangsbo J (2003) Muscle interstitial potassium kinetics during intense exhaustive exercise: effect of previous arm exercise. Am J Physiol Regul Integr Comp Physiol 285:R143–R148PubMedGoogle Scholar
  57. Paganini AT, Foley JM, Meyer RA (1997) Linear dependence of muscle phosphocreatine kinetics on oxidative capacity. Am J Physiol 272:C501–C510PubMedGoogle Scholar
  58. Peacock O, Tjønna AE, James P, Wisløff U, Welde B, Böhlke N, Smith A, Stokes K, Cook C, Sandbakk O (2012) Dietary nitrate does not enhance running performance in elite cross-country skiers. Med Sci Sports Exerc 44:2213–2219PubMedCrossRefGoogle Scholar
  59. Poveda JJ, Riestra A, Salas E, Cagigas ML, López-somoza C, Amado JA, Berrazueta JR (1997) Contribution of nitric oxide to exercise-induced changes in healthy volunteers: effects of acute exercise and long-term physical training. Eur J Clin Invest 27:967–971Google Scholar
  60. Rampinini E, Sassi A, Azzalin A, Castagna C, Menaspà P, Carlomagno D, Impellizzeri FM (2010) Physiological determinants of Yo–Yo intermittent recovery tests in male soccer players. Eur J Appl Physiol 108:401–409PubMedCrossRefGoogle Scholar
  61. Richardson RS, Leigh JS, Wagner PD, Noyszewski EA (1999) Cellular PO2 as a determinant of maximal mitochondrial O2 consumption in trained human skeletal muscle. J Appl Physiol 87:325–331PubMedGoogle Scholar
  62. Sobko T, Marcus C, Govoni M, Kamiya S (2010) Dietary nitrate in Japanese traditional foods lowers diastolic blood pressure in healthy volunteers. Nitric Oxide 22:136–140PubMedCrossRefGoogle Scholar
  63. Stamler JS, Meissner G (2001) Physiology of nitric oxide in skeletal muscle. Physiol Rev 81:209–237PubMedGoogle Scholar
  64. Totzeck M, Hendgen-Cotta UB, Rammos C, Frommke LM, Knackstedt C, Predel HG, Kelm M, Rassaf T (2012) Higher endogenous nitrite levels are associated with superior exercise capacity in highly trained athletes. Nitric Oxide 27:75–81PubMedCrossRefGoogle Scholar
  65. Tsintzas K, Williams C (1998) Human muscle glycogen metabolism during exercise. Effect of carbohydrate supplementation. Sports Med 25:7–23PubMedCrossRefGoogle Scholar
  66. Vanhatalo A, Bailey SJ, Blackwell JR, DiMenna FJ, Pavey TG, Wilkerson DP, Benjamin N, Winyard PG, Jones AM (2010a) Acute and chronic effects of dietary nitrate supplementation on blood pressure and the physiological responses to moderate-intensity and incremental exercise. Am J Physiol Regul Integr Comp Physiol 299:R1121–R1131PubMedCrossRefGoogle Scholar
  67. Vanhatalo A, Fulford J, DiMenna FJ, Jones AM (2010b) Influence of hyperoxia on muscle metabolic responses and the power-duration relationship during severe-intensity exercise in humans: a 31P magnetic resonance spectroscopy study. Exp Physiol 95:528–540PubMedCrossRefGoogle Scholar
  68. Vanhatalo A, Fulford J, Bailey SJ, Blackwell JR, Winyard PG, Jones AM (2011) Dietary nitrate reduces muscle metabolic perturbation and improves exercise tolerance in hypoxia. J Physiol 589:5517–5528PubMedGoogle Scholar
  69. Veale JP, Pearce AJ, Carlson JS (2010) The Yo–Yo intermittent recovery test (level 1) to discriminate elite junior Australian football players. J Sci Med Sport 13:329–331PubMedCrossRefGoogle Scholar
  70. Vernillo G, Silvestri A, Torre AL (2012) The Yo–Yo intermittent recovery test in junior basketball players according to performance level and age group. J Strength Cond Res 26:2490–2494PubMedCrossRefGoogle Scholar
  71. Wilkerson DP, Hayward GM, Bailey SJ, Vanhatalo A, Blackwell JR, Jones AM (2012) Influence of acute dietary nitrate supplementation on 50 mile time trial performance in well-trained cyclists. Eur J Appl Physiol 112:4127–4134Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Lee J. Wylie
    • 1
  • Magni Mohr
    • 1
  • Peter Krustrup
    • 1
  • Sarah R. Jackman
    • 1
  • Georgios Ermιdis
    • 1
  • James Kelly
    • 1
  • Matthew I. Black
    • 1
  • Stephen J. Bailey
    • 1
  • Anni Vanhatalo
    • 1
  • Andrew M. Jones
    • 1
    Email author
  1. 1.Sport and Health Sciences, College of Life and Environmental Sciences, University of ExeterExeterUK

Personalised recommendations