The complex reactions controlling the formation of tobacco-specific nitrosamines (TSNA) in the curing of tobacco have caused difficulty in developing a curing procedure to decrease nitrosamine levels in tobacco. Previous studies have shown that there was a relationship between TSNA and alkaloid levels, nitrate levels or nitrite levels. Many of the early studies focussed only on the relationship of one plant constituent and TSNA accumulation. In a more recent integrated study, we have shown there was a relationship between nitrite and TSNA but there was a more positive relationship between the secondary amine pyridine alkaloid precursors and TSNA from the same air-cured lamina. Data from this most recent study show secondary amine alkaloids are generally in 10² to 10³fold excess of nitrite in cured leaf. This result implies that nitrite, but not secondary pyridine alkaloids, is limiting nitrosamine accumulation. To investigate the premise that nitrite limits nitrosamine accumulation, an aqueous solution of nitrite was added to a dark air-cured tobacco in such a manner the moisture content was increased to 40%. The head space and tobacco tissue were sampled at l, 2, 3, 5, 7, 14, 21, and 28 days. Oxides of nitrogen were measured in the head space and freeze dried leaf tissue were analyzed for individual TSNA, nitrite, nitrate and alkaloids. Nitrite concentration was maximum at the first sampling date and was at a minimum by day 14. Maximum rate of accumulation of TSNA occurred during the first day. The conclusion drawn from this study is nitrite levels are limiting nitrosamine accumulation.