Purpose: Hydrogen-rich water (HRW) has been shown to have an antifatigue effect. This study assessed up-hill running performance, as well as physiological and perceptual responses after supplementation with 1680 mL HRW between 24 h and 40 min before running, in athletes of heterogeneous running ability. Methods: Sixteen males (mean [SD] age 31.6 [8.6] y, VO2max 57.2 [8.9] mL·kg−1·min−1, body fat 13.4% [4.4%]) participated in this study. Using a randomized, double-blind, placebo-controlled crossover design, participants consumed either HRW or placebo prior to performing two 4.2-km up-hill races separated by a week. Race time (RT), average race heart rate, and immediately postrace rating of perceived exertion were assessed. Results: After analysis of data for all runners, HRW effect was unclear (−10 to 7 s, 90% confidence interval) for RT, likely trivial for heart rate (−2 to 3 beats·min−1), and likely trivial for postrace rating of perceived exertion (−0.1 to 1.0). A possible negative correlation was found between RT differences and average RT (r = −.79 to −.15). HRW for the 4 slowest runners (RT = 1490  s) likely improved the RT (−36 to −3 s), whereas for the 4 fastest runners (RT = 1069  s) the performance effect of HRW was unclear (−10 to 26 s). Conclusions: HRW intake had an unclear antifatigue effect on performance in terms of mean group values. However, it appears that the magnitude of the antifatigue effect of HRW on performance depends on individual running ability.