Abstract: In order to understand the content and composition characteristics of amino acids in stems of flue-cured tobacco leaves, 20 kinds of amino acids in flue-cured tobacco stems and corresponding lamina samples including upper/middle/lower stalk position from 18 representative domestic areas, were analyzed. With statistical analysis, the differences of amino acid contents between tobacco stems and laminas were compared. The correlation between amino acid contents of tobacco stems and corresponding laminas was analyzed, and the content differences of amino acid among tobacco stems of different stalk position were investigated. The results showed that the total amount of amino acids in tobacco stems was slightly higher than that in laminas, and the contents of 11 amino acids such as isoleucine, tyrosine and aspartic acid in tobacco stems were significantly higher than those in corresponding laminas, while the contents of 4 amino acids such as tryptophan and histidine in tobacco stems were significantly lower than those in laminas; the variability of amino acid contents among different tobacco stems was greater than that among laminas; except valine, threonine and cystine, other amino acids in tobacco stems were significantly related to laminas positively; the amino acid contents of upper tobacco stems were higher than that of middle and lower stems. In summary, there was a positive correlation between amino acid content of tobacco stem and corresponding laminas, but there was significant difference in amino acid composition.