Abstract: Several pre-workout supplements contain niacin, although the exercise performance effects of niacin are poorly understood. The purpose of the present study was to examine the performance effects of niacin versus caffeine as a pre-workout supplement. Twenty-five untrained males were recruited to complete three identical ramped aerobic cycling exercise trials. Participants were administered caffeine (CA) at 5 mg/kg body weight, 1000 mg niacin (NI), or a methylcelluloce placebo (PL) supplement prior to each trial. NI treatment induced significantly higher respiratory exchange ratio (RER) during exercise compared to the CA treatment, but not the PL treatment (PL=0.87±0.08, NI=0.91±0.08, CA=0.87±0.08; p=0.02). Similarly, exercise time to exhaustion (in minutes) was significantly different between the NI treatment and the CA treatment, but not the PL treatment (PL=27.45±4.47, NI=26.30±4.91, CA=28.76±4.86; p<0.01). Habitual caffeine use (p=0.16), habitual aerobic exercise (p=0.60), and habitual resistance exercise (p=0.10) did not significantly affect RER. Similarly, habitual caffeine use (p=0.72), habitual aerobic exercise (p=0.08), and habitual resistance exercise (p=0.39) did not significantly affect total work performed. The elevated RER and decreased time to exhaustion in the NI treatment suggests limited lipid availability during exercise and impaired exercise performance.