Heat-stabilized protein is an important index to measure the quality of malt. In order to explore the content of heat-stabilized protein, the composition characteristics of protein Z and the influencing conditions in barley grains and malt, three hulless barleys (Liulengzi changmang baiqingke, Qingpi qingke and 9820) and one hulled barley (Gairdner ) were used to analyze and identify the heat-stabilized protein of hulless barley grains and malt. Then the content of the heat-stabilized protein of hulless barley and the composition characteristics of protein Z were studied in different ecological environments. The barley lines with excellent brewing quality were selected. The results showed that the optimal barley germination conditions (20℃, 72h and PH 5) enabled the accumulation of heat-stabilized protein and protein Z in germination and kinling. The hulless barleys, which were planted and harvested at three locations (Xining, Huangyuan and Haiyan) with different climatic conditions, were tested for the content of grain. A lowest bud heat-stabilized protein, but the highest heat-stabilized protein and protein Z after kinling, were observed from the grains in Xining. Meanwhile, fifteen excellent resources with high content of heat-stable protein were identified by testing 150 hulless barley germplasm resources. Thus, the results gained from this study will provide theoretical basis in breeding for hulless malting barley varieties with the qualified indexes.