Black highland barley is rich in a variety of nutrients and bioactive ingredients, and hence of important research significance and development value. This work investigated the fermentation characteristics of yeast in the wort having different proportions of black barley malt and barley malt (10:0, 8:2, 6:4, 4:6, 2:8, and 0:10 m/m; referred to as HA, HB, HC, HD, HE and HF, respectively) and the nutritional composition, quality, and sensory characteristics of the resulting beer. The results indicated that the addition of black barley malt significantly decreased the pH value, fermentability, and ethanol yield (P<0.05) thereby affecting yeast fermentation performance. An increase in the proportion of black barley malt resulted in an increase, in the β-glucan and anthocyanin contents, turbidity, and chroma of the resulting beer while the polyphenol content increased up to 105.31 mg/L in HB beer sample. The HE and HD beer samples exhibited high foaming ability , while the HB, HC, HD, and HE beer samples demonstrated good foam retention capacity. A total of 54 types of flavor substances were detected from the six beer samples. The addition of black barley malt caused a reduction in the type and concentration of alcohols while increasing the ester compounds. Around 32 esters were identified in the HB and HC beer samples. PCA analysis revealed significant differences in the flavor components of the different samples indicating that the addition of black barley malt could significantly improve the flavor composition of beer. The HB and HC beer samples obtained higher scores in sensory analysis. In summary, addition of black barley malt at 20% to 40% (m/m), resulted in beer with excellent quality, flavor, and sensory characteristics.