Abstract: In this study, nine varieties of walnut nuts were used as test materials to determine the amino acid composition and content, calculate their nutritional value scores, and conduct principal component analysis and cluster analysis to comprehensively evaluate the nutritional value of amino acids from multiple perspectives. The results showed that 17 kinds of amino acids were detected in different varieties of walnuts, with a total amount of 15.33~20.45 g/100 g. Flavor amino acids and medicinal amino acids accounted for 55.25%~58.04% and 63.56%~70.16% of the total amino acids, respectively. Various methods including amino acid scoring, chemical scoring and amino acid ratio coefficient all determined that lysine was the first limiting amino acid. The comprehensive performance and high scores of QK, XL3, XL, LG, and QX varieties were demonstrated by principal component analysis based on amino acid composition and content. Higher essential amino acid index and biological value, rich flavor and functional amino acid contents, and relatively good quality were observed in walnut varieties QK, QX, XL, XL3, and LG through cluster analysis based on nutritional evaluation. Comprehensive analysis showed that QK, XL and XL3 were excellent varieties with high nutritional value of amino acids and were suitable for promotion of planting and processing utilization.