Abstract: This study used walnut male flowers collected from 11 distinct regions to analyze their amino acid composition and content using an automatic amino acid analyzer. Correlation analysis, principal component analysis (PCA) and cluster analysis were conducted for a comprehensive evaluation. The results indicated that all samples contained 16 amino acids, including 7 essential amino acids (lysine, methionine, threonine, isoleucine, leucine, valine, tyrosine), 2 semi-essential amino acids (histidine, arginine) and 7 non-essential amino acids (glycine, alanine, proline, serine, cystine, aspartic acid, glutamic acid). The total amino acid content across regions ranged from 10.63% to 16.78%. The sample from Hebei exhibited the highest tota lamino acid content (16.78%), as well as the highest levels of flavor-related and seasoning amino acids (26.88%) and medicinal amino acids (11.88%), suggesting that walnut male flowers from this region possessed considerable potential for application in food research and development. PCA revealed that the first three principal components collectively accounted for 96.51% of the total variance, effectively capturing the comprehensive amino acid profile. The top four regions based on comprehensive scores were Hebei, Henan, Jiangsu and Shandong. Cluster analysis grouped the 11 regions into three categories, consistent with the PCA findings, and accurately reflected the regional differences in amino acid composition, flavor characteristics and functional properties. These findings provide a valuable theoretical basis for selecting suitable planting regions and guiding the processing and utilization of walnut male flowers.