A better understanding of amino acids (AA) characteristics is beneficial for breeding, processing and storage of sugarcane. The differences and classification in AA of 10 different sugarcane varieties (S1-S10) were investigated in this study. Varieties affected components and concentrations of free AA (p < 0.05) significantly. The total AA of sugarcanes ranged from 51.44 to 781.61 mg/L, while the content of essential amino acids (EAA) was in the range of 0.72–60.98 mg/L. Glu (glutamic acid) was the most abundant free amino acids in all varieties of sugarcane except S4. Even though S6 had the highest concentration of medicinal AA (781.61 mg/L), the highest proportion of medicinal AA in total AA was found in S5 (81.76%). S6 got the highest comprehensive score in amino acids based on the results of Principal Component Analysis (PCA). The cluster analysis roughly divided the sugarcanes into 2 groups. Group Ⅰ including S1, S6, S2 and S5 got positive scores in PCA might had great edible and medicinal development and utilization value.