Glycerin is applied to cigarette tobacco at levels in the range of about 1-5% to improve moisture holding characteristics of tobacco and act as a surface active agent for flavor application. Neat material pyrolysis studies, smoke chemistry and biological activity studies (bacterial mutagenicity, cytotoxicity, in vivo micronucleus, and sub-chronic rodent inhalation) with mainstream smoke, or mainstream smoke preparations from cigarettes containing various target levels (5%, 10%, and 15%) of the glycerin were performed to provide data for an assessment of the use of glycerin as a cigarette tobacco ingredient. The actual levels of glycerin in the respective test cigarettes were determined to be 3.2%, 6.2% and 8.4% after cigarette production. At simulated tobacco burning temperatures up to 900 degrees C, neat glycerin did not pyrolyze extensively suggesting that glycerin would transfer intact to mainstream smoke (smoke was not analyzed for glycerin in this study). On a tar basis, nicotine in smoke was significantly decreased at 10% and 15% glycerin while water was increased at all addition levels. Addition of 10% or 15% glycerin also resulted in a statistically significant increase in acrolein (9%) and a decrease in acetaldehyde, propionaldehyde, aromatic amines, nitrogen oxides, tobacco specific nitrosamines, and phenols. Addition of 5% glycerin produced the same decrease in smoke constituents as the 10% and 15% groups but there was no concomitant increase in acrolein. Biological tests indicated no relevant differences in the genotoxic or cytotoxic potential of either mainstream smoke (or smoke preparations) from cigarettes with added glycerin compared to control cigarettes. Cigarette smoke atmosphere dilution, coupled with the lower nicotine delivery in the test cigarettes containing glycerin resulted in a lower nicotine delivery to the glycerin cigarette smoke exposed rats of the 90-day inhalation study. Smoke atmosphere acrolein was also reduced in a concentration-related manner. Incorporation of glycerin at target levels up to 15% did not produce any adverse effects in rats exposed for 90-days. The major observation in the study was a reduced biological activity of the smoke as indicated by a reduction in the severity and/or incidence of focal macrophage accumulation in the lungs and goblet cell hyperplasia/hypertrophy in the nose (level 1), and goblet cell staining depletion in the nose (level 1). The results of these studies with glycerin applied to cigarette tobacco suggest that adding glycerin to cigarette tobacco at typical use levels does not adversely alter the smoke chemistry or biological effects normally associated with exposure to mainstream cigarette smoke.