In order to explore the differences in the quality of hulless barley (Hordeum vulgare L. var. nudum Hook.f.) varieties in the Qinghai-Tibet Plateau, establish the quality evaluation index system of hulless barley, and regionalize the ecological adaptability of hulless barley quality, we used 15 main hulless barley varieties in the Qinghai-Tibet Plateau as experimental materials and measured thirteen quality indexes such as total starch, crude protein, ether extract and β-glucan. One-way analysis of variance, correlation analysis, principal component analysis and cluster analysis were used to analyze the measured indexes. The results showed that: (1) among the 13 quality indexes, 11 indexes had significant differences among the 15 tested varieties, and 12 indexes had significant differences in the 8 planting zones. There was a significant positive correlation between the total flavonoid content and the total polyphenol content, and the β-glucan content. Based on the performance of different varieties in different ecological regions, the contents of amylopectin, amylose, total flavonoids, β-glucan and total polyphenols were the greatest influence on the comprehensive quality of hulless barley. (2) The 15 tested varieties were divided into 4 categories by cluster analysis, and the 8 regions were divided into 3 ecological zones. The varieties with better quality in each ecological zone were Kunlun 18, Kunlun 19, Kunlun 15, Ganqing 8 and Ganqing 4. (3) These results showed that 13 indicators could be converted into 5 principal components, and the cumulative variance contribution rates were 85.16% and 94.40%, respectively. The variety with the highest comprehensive evaluation score was Kunlun 18, and the area with the best quality was Guinan, Qinghai province.