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Updated: Jan 18, 2026

Assessing Energy Substrate Oxidation In Vitro with 14CO2 Trapping
Published on: March 23, 2022
Substrate Oxidation Does Not Influence Middle Distance Running Performance: A Randomized Controlled Crossover Trial
Alex Buga1, Jeffrey D Buxton2, Emma Plank2
1Department of Human Sciences, The Ohio State University, Columbus, OH 43210, USA.
Objective:
Recent work has challenged the notion that preferred substrate oxidation is a key determinant of exercise performance. This investigation tested middle-distance running performance, in the fed state, to control for glycogen and exercise-induced hypoglycemia (EIH) confounders.
Methods:
In a randomized crossover fashion, all while controlling dietary intake, activity, and body weight, recreational distance runners completed either a 5K (n = 15; VO2max: 58.3 ± 6.2 mL/kg/min) or a 10K (n = 15; VO2max: 54.51 ± 5.9 mL/kg/min) middle-distance run after consuming isocaloric low-carbohydrate high-fat (LCHF) and high-carbohydrate low-fat (HCLF) pre-exercise meals. Time trial (TT) performance (sec), carbohydrate/fat substrate oxidation, blood metabolites, heart rate (HR), ratings of perceived exertion (RPE), and subjective fullness and thirst were measured throughout.
Results:
LCHF pre-exercise nutrition reliably altered substrate oxidation and metabolite profiles compared to HCLF, evidenced by significant increases in fat oxidation (77% higher) and reductions in RER (5% lower), with corresponding shifts in carbohydrate oxidation. Despite distinct preferred substrate oxidation profiles during exercise, the 5 and 10 km TT performances were similar between conditions (p = 0.646/p = 0.118). RER was significantly lower (p = 0.002) after the LCHF condition compared to HCLF. Capillary R-βHB increased modestly after LCHF, while blood glucose increased after HCLF only. The LCHF meal was 35% more filling than the HCLF meal. Preferred substrate oxidation did not significantly modulate middle-distance running performance.
Conclusion:
This work supports recent findings that substrate oxidation is not a primary determinant of aerobic performance, as previously conceived.
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