Abstract: A better understanding of the effects of stalk cutting time on free amino acid is necessary to further improve the leaf quality of burley tobacco. The objective of this study was to investigate whether there were differences in the free amino acid contests of three stalk leaves among the harvest time of 3, 4, 5 and 6 weeks after topping in burley tobacco. The results showed that out of 16 free amino acid types, three free amino acids, Glu, Asp, Pro, had a ratio of 63.60% to 74.33% to total amino acids. The total contents of other 13 types were lower, about 25.67% to 36.40%. Significant differences were observed in the content of free amino acids among different stalk leaves. The highest and lowest values were found in the harvest time of 3 and 5 weeks after topping in lugs, respectively. Although it had the lowest contents of free amino acids in the treatments of the harvest time of 6 weeks after topping in both cutters and leaves, the highest values were observed in the treatments of the harvest time of 4 and 5 weeks after topping, respectively. Average across three stalk leaves, it was greatest and lowest in the treatments of the harvest time of 4 and 5 weeks after topping, respectively. The impact of stalk cutting time on free amino acids was greatest in lugs, followed by leaves and cutters. Stalk-cut period also had a higher effect on the contents of ser, his, ile than those of phe, glu and FAA. It was concluded that the best harvest time to obtain good quality in burley tobacco was 3, 4 and 5 weeks after topping for lugs, cutters and leaves, respectively.