Accumulation of nitrosamines in tobacco and conditions which influence their formation have been of recent interest. A study has been completed to determine if there are differences in tobacco-specific nitrosamines (TSNA) levels from different tobacco varieties. Three tobacco varieties, KY 14, G28, and KY 171, which represents burley, flue-cured and dark air-cured tobacco, respectively, were grown at Lexington, Kentucky in 1988 and 1989 using cultural practices for the production of burley tobacco. The tobaccos were stalk cut and air-cured in both controlled curing (24.degree./70% RH) and in a conventional curing barn. At end of curing, three replicate samples were taken for top, middle and bottom of the plant for each variety. The leaves were separated into lamina and mid-vein, ground and stored at -40.degree.C until analyses. The tobacco samples were analyzed for individual alkaloids, nitrate-nitrogen, nitrite-nitrogen and the individual tobacco-specific nitrosamines. Analyses of the data indicate there is significant differences in nitrosamines and specific tobacco varieties, whereas there was no stalk position effect. There was a positive correlation between the concentration of nitrite and accumulation of tobacco-specific nitrosamines. These data indicate that accumulation of tobacco-specific nitrosamines was dependent on the accumulation of nitrite during curing and storage of tobacco.