Introduction: The effect of branched-chain amino acids (BCAA) or glucose supplementation on exercise performance were investigated in the normal atmospheric pressure with normoxia condition in previous studies. No studies had been investigated the effect of BCAA and glucose supplementation before exercise on exercise performance in normal atmospheric pressure with hypoxia condition. The purpose of this study was to examine the effect of ingestion of glucose and BCAA before exercise on endurance performance and physiological response in an acute hypoxia condition. Methods: Seven healthy college male subjects completed 4 experimental trials in a random order. They ingested either 12 g of BCAAs (BCAA trial), 12g branched-chain amino acids plus 75g glucose (BCAA+GLU trial), 75g glucose (GLU trial) or placebo (PLA trial) 1 hour before exercise in a hypoxia chamber. Ten minutes before exercise, the subjects were asked to enter the hypoxia chamber for adaption the hypoxia environment. Subjects exercised at 50% V02max on cycle erogmeter for 90 minutes. After rest for 1 minute, the exercise intensity then increased to the intensity of 85% V02max till exhaustion. Results: The results showed that there were no differences on exercise performance between 4 trials (85% V02max to fatigue: BCAA 353.4 ± 84.2 seconds, BCAA+GLU 340.9 ± 90 seconds, GLU 407 ± 77.3 seconds, and PLA 291.6 ± 42.9). The ratio of plasma tryptophan (TRP) to BCAA concentrations were significantly lower in BCAA and BCAA+GLU trials than GLU and PLA trials (p < 0.008). Plasma insulin concentrations were significant higher in BCAA+GLU compared to BCAA (p = 0.008) and PLA (p = 0.004). Plasma glucose concentrations were significant higher in BCAA+GLU than that of BCAA (p = 0.02). There were no differences in plasma NH3 concentration, fat oxidation rates and carbohydrate oxidation rates during exercise between 4 trials. Conclusion: The current results suggested ingestion carbohydrate and BCAAs did not alter endurance performance in acute hypoxia environment.