2S-hesperidin is a flavanone (flavonoid) found in high concentrations in citrus fruits. It has an antioxidant and anti-inflammatory effect, improving performance in animals. This study investigated the effects of chronic intake of an orange extract (2S-hesperidin) or placebo on aerobic-anaerobic and metabolic performance markers in amateur cyclists. A double-blind, randomized, placebo-controlled trial was carried out between late September and December 2018. Forty amateur cyclists were randomized into two groups: one taking 500mg/day 2S-hesperidin and other taking 500 mg/day placebo (microcellulose) for 8 weeks. All participants completed the study. Performance and metabolic aerobic-anaerobic markers were measured using incremental and rectangular tests by indirect calorimetry. The anaerobic power was determined using Wingate tests. After 8 weeks supplementation, there was a significant increase in the incremental test in estimated functional threshold power (FTP) (3.23%; p≤0.05) and maximum power (2.68%; p≤0.05) with 2S-Hesperdin compared to placebo. In the rectangular test, there was a significant decrease in VO2 (-8.26%; p≤0.01) and VO2R (-8.88%; p≤0.01) at VT2 in placebo; however, there were no significant differences between groups. In the Wingate test, there was a significant increase (p≤0.05) in peak and relative power in both groups, but without significant differences between groups. Supplementation with an orange extract (2S-hesperdin) 500mg/day improves estimated FTP and maximum power performance in amateur cyclists.