We hypothesized that acute dietary nitrate (NO3-) provided as concentrated beetroot juice supplement would improve endurance running performance of well-trained runners in normobaric hypoxia. Ten male runners (mean (SD): sea level V̇O2max 66 (7) mL.kg-1.min-1, 10 km personal best 36 (2) min) completed incremental exercise to exhaustion at 4000 m and a 10 km treadmill time trial at 2500 m simulated altitude on separate days, after supplementation with ~7 mmol NO3- and a placebo, 2.5 h before exercise. Oxygen cost, arterial oxygen saturation, heart rate and ratings of perceived exertion (RPE) were determined during the incremental exercise test. Differences between treatments were determined using means [95% confidence intervals], paired sample t-tests and a probability of individual response analysis. NO3- supplementation increased plasma [nitrite] (NO3-, 473 (226) nM vs. placebo, 61 (37) nM, P <0.001) but did not alter time to exhaustion during the incremental test (NO3-, 402 (80) s vs. placebo 393 (62) s, P = 0.5) or time to complete the 10 km time trial (NO3-, 2862 (233) s vs. placebo, 2874 (265) s, P = 0.6). Further, no practically meaningful beneficial effect on time trial performance was observed as the 11 [-60 to 38] s improvement was less than the a priori determined minimum important difference (51 s), and only three runners experienced a ´likely, probable´ performance improvement. NO3- also did not alter oxygen cost, arterial oxygen saturation, heart rate or RPE. Acute dietary NO3- supplementation did not consistently enhance running performance of well-trained athletes in normobaric hypoxia.