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<h4>Background</h4>Submaximal endurance exercise has been shown to cause elevated gastrointestinal permeability, injury, and inflammation, which may negatively impact athletic performance and recovery. Preclinical and some clinical studies suggest that flavonoids, a class of plant secondary metabolites, may regulate intestinal permeability and reduce chronic low-grade inflammation. Consequently, the purpose of this study was to determine the effects of supplemental flavonoid intake on intestinal health and cycling performance.<h4>Materials and methods</h4>A randomized, double-blind, placebo-controlled crossover trial was conducted with 12 cyclists (8 males and 4 females). Subjects consumed a dairy milk-based, high or low flavonoid (490 or 5 mg) pre-workout beverage daily for 15 days. At the end of each intervention, a submaximal cycling trial (45 min, 70% VO2max) was conducted in a controlled laboratory setting (23°C), followed by a 15-minute maximal effort time trial during which total work and distance were determined. Plasma samples were collected pre- and post-exercise (0h, 1h, and 4h post-exercise). The primary outcome was intestinal injury, assessed by within-subject comparison of plasma intestinal fatty acid-binding protein. Prior to study start, this trial was registered at ClinicalTrials.gov (NCT03427879).<h4>Results</h4>A significant time effect was observed for intestinal fatty acid binding protein and circulating cytokines (IL-6, IL-10, TNF-α). No differences were observed between the low and high flavonoid treatment for intestinal permeability or injury. The flavonoid treatment tended to increase cycling work output (p = 0.051), though no differences were observed for cadence or total distance.<h4>Discussion</h4>Sub-chronic supplementation with blueberry, cocoa, and green tea in a dairy-based pre-workout beverage did not alleviate exercise-induced intestinal injury during submaximal cycling, as compared to the control beverage (dairy-milk based with low flavonoid content).