Ascending to high-altitude of non-high-altitude natives showed rapidly declined physical ability, but some with high-altitude adaptability were still able to maintain a high level. Our goal was to find urine biomarkers that could be used to assess people for high-altitude hypoxia adaptation with high stamina. Blood routine tests were done for HH (High-altitude with high stamina) and HL (High-altitude with low stamina) groups at high altitude and at sea level. By contrasting urine peptidome profiling between the HH and HL groups, urine biomarkers were discovered. Blood routine tests showed that WBC, LYM, and PLT concentrations were considerably greater in HH than in HL. Four proteins have significantly different levels of expression between HH and HL, according to urine peptidome profiling. Expression of these proteins were tested by ELISA in HH and HL samples. Our study provided a novel approach for identifying potential biomarkers for screening people who can adapt to high altitudes with high stamina.