We determined if high cadences, during a prolonged cycling protocol with varying intensities (similar to race situations) decrease performance compared to cycling at a lower, more energetically optimal, cadence. Eight healthy, competitive male road cyclists (35 ± 2 yr) cycled for 180 min at either 80 or 100 rpm (randomized) with varying intensities of power outputs corresponding to 50, 65 and 80% of VO₂max. At the end of this cycling period, participants completed a ramped exercise test to exhaustion at their preferred cadence (90 ± 7 rpm). There were no cadence differences in blood glucose, respiratory exchange ratio or rate of perceived exertion. Heart Rate, VO₂ and blood lactate were higher at 100 rpm vs. 80 rpm. The total energy cost while cycling during the 65% and 80% VO₂max intervals at 100 rpm (15.2 ± 2.7 and 19.1 ± 2.5 kcal∙min^-1, respectively) were higher than at 80 rpm (14.3 ± 2.7 and 18.3± 2.2 kcal∙min^-1, respectively) (p < 0.05). Gross efficiency was higher at 80 rpm vs. 100 rpm during both the 65% (22.8 ± 1.0 vs. 21.3 ± 4.5%) and the 80% (23.1 vs. 22.1 ± 0.9%) exercise intensities (P< 0.05). Maximal power during the performance test (362 ± 38 watts) was greater at 80 rpm than 100 rpm (327 ± 27 watts) (p < 0.05). Findings suggest that in conditions simulating those seen during prolonged competitive cycling, higher cadences (i.e., 100 vs. 80 rpm) are less efficient, resulting in greater energy expenditure and reduced peak power output during maximal performance.