¹⁵ N-labeled dietary nitrate supplementation increases human skeletal muscle nitrate concentration and improves muscle torque production

Aim: Dietary nitrate (NO₃ ^- ) supplementation increases nitric oxide bioavailability and can enhance exercise performance. We investigated the distribution and metabolic fate of ingested NO₃ ^- at rest and during exercise with a focus on skeletal muscle.

Methods: In a randomized, crossover study, 10 healthy volunteers consumed 12.8 mmol ¹⁵ N-labeled potassium nitrate (K¹⁵ NO₃ ; NIT) or potassium chloride placebo (PLA). Muscle biopsies were taken at baseline, at 1- and 3-h post-supplement ingestion, and immediately following the completion of 60 maximal intermittent contractions of the knee extensors. Muscle, plasma, saliva, and urine samples were analyzed using chemiluminescence to determine absolute [NO₃ ^- ] and [NO₂ ^- ], and by mass spectrometry to determine the proportion of NO₃ ^- and NO₂ ^- that was ¹⁵ N-labeled.

Results: Neither muscle [NO₃ ^- ] nor [NO₂ ^- ] were altered by PLA. Following NIT, muscle [NO₃ ^- ] (but not [NO₂ ^- ]) was elevated at 1-h (from ~35 to 147 nmol/g, p < 0.001) and 3-h, with almost all of the increase being ¹⁵ N-labeled. There was a significant reduction in ¹⁵ N-labeled muscle [NO₃ ^- ] from pre- to post-exercise. Relative to PLA, mean muscle torque production was ~7% greater during the first 18 contractions following NIT. This improvement in torque was correlated with the pre-exercise ¹⁵ N-labeled muscle [NO₃ ^- ] and the magnitude of decline in ¹⁵ N-labeled muscle [NO₃ ^- ] during exercise (r = 0.66 and r = 0.62, respectively; p < 0.01).

Conclusion: This study shows, for the first time, that skeletal muscle rapidly takes up dietary NO₃ ^- , the elevated muscle [NO₃ ^- ] following NO₃ ^- ingestion declines during exercise, and muscle NO₃ ^- dynamics are associated with enhanced torque production during maximal intermittent muscle contractions.