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This study aimed to investigate carbohydrate (CHO) mouth rinse response on neuromuscular activity, fuel oxidation rates, and cycling performance with different initial levels of endogenous CHO availability.In a double-blind, randomized placebo-controlled design, eight males completed six experimental mouth rinse trials: CHO (6.4% maltodextrin) or placebo solution in a fed state (FED), 12-h fasted state (FAST), or a combined exercise-depleted muscle glycogen and 12-h fasted state (DEP). Trials consisted of 30-min cycling at 90% of gas exchange threshold, followed by a 20-km cycling time trial. Plasma lactate, plasma glucose, oxygen uptake, and EMG activity were measured, and CHO and fat oxidation rates were calculated.CHO mouth rinse maintained higher plasma glucose levels as the constant load exercise progressed (P = 0.023). The reduced EMG activity in the DEP condition with the placebo during constant load exercise was ameliorated with CHO mouth rinse (P < 0.01). Furthermore, the power output and the EMG activity throughout the 20-km time trial were reduced in the DEP condition with placebo but were both restored with CHO mouth rinse (P < 0.05). Time trial performance was only improved with CHO in the DEP compared with the corresponding placebo (P < 0.05), and no differences between supplements were observed in the FED or FAST states. Analyses of the qualitative inference showed “benefit very likely” of CHO mouth rinse on exercise performance in DEP, “possibly benefit” in FAST, and “negligible or trivial” in FED. CHO mouth rinse had no effect on CHO and fat oxidation rates in either exercise mode.The CHO mouth rinse influences exercise performance when endogenous CHO availability is low, and an enhanced central motor drive is potentially the main influencing mechanism.