Several studies have suggested that the most economical cadence in cycling increases with increasing workload. However, none of these studies have been able to demonstrate this relationship with experimental data. The purpose of this study was to test the hypothesis that the most economical cadence in elite cyclists increases with increasing workload and to explore the effect of cadence on performance. Six elite road cyclists performed submaximal and maximal tests at four different cadences (60, 80, 100 and 120 rpm) on separate days. Respiratory data was measured at 0, 50, 125, 200, 275 and 350 W during the submaximal test and at the end of the maximal test. The maximal test was carried out as an incremental test, conducted to reveal differences in maximal oxygen uptake and time to exhaustion (short-term performance) between cadences. The results showed that the lowest oxygen uptake, i.e. the best work economy, shifted from 60 rpm at 0 W to 80 rpm at 350 W ( P<0.05). No difference was found in maximal oxygen uptake among cadences ( P>0.05), while the best performance was attained at the same cadence that elicited the best work economy (80 rpm) at 350 W ( P<0.05). This study demonstrated that the most economical cadence increases with increasing workload in elite cyclists. It was further shown that work economy and performance are related during short efforts (approximately 5 min) over a wide range of cadences.