Abstract: In this study, 17 amino acids were analyzed by amino acid analyzer in 110 honey samples from three different monofloral honeys, i.e.acacia honey, linden honey and rape honey in China, and the honey variety discrimination was performed based on amino acid content combined with chemometrics. The results showed that the content of hydrolyzed amino acids was higher in rape honey than in acacia and linden honey, and the content of 16 amino acids showed significant differences among the honey varieties. The results of principal component analysis (PCA) revealed that the botanical origins of honey samples clustered according to their amino acid content. The results of least squares discriminant analysis (PLS-DA) showed that rape honey could be distinguished from acacia honey and linden honey. Linear discriminant analysis (LDA) showed that the overall discriminant rate of the three honeys was 92.7%, and the discriminant rate of rape honey was 92.3%. This study would provide data support and reference for the classification and identification of rape honey, acacia honey and linden honey.