Background The widespread use of smokeless tobacco, particularly in India, is a major public health concern, contributing to increased oral cancer cases. Tobacco pouch keratosis, an early sign of oral mucosal damage, is commonly observed in regular smokeless tobacco users. This study examines the impact of smokeless tobacco on salivary antioxidant systems, comparing individuals with and without tobacco pouch keratosis to healthy controls. Materials and methods A total of 63 participants were divided into three groups: smokeless tobacco users without oral lesions (group 1, n=23), users with tobacco pouch keratosis (group 2, n=20), and healthy controls (group 3, n=20). Salivary parameters, including flow rate, pH, and total antioxidant capacity (measured by the ferric reducing antioxidant power [FRAP] assay), were analyzed. A descriptive analysis was conducted for both continuous and categorical variables. For inferential statistics, the Mann-Whitney U test was used to compare the duration and frequency of tobacco use. A one-way ANOVA, followed by Tukey's post-hoc test, was applied to compare the mean salivary flow rate and salivary pH levels. Spearman's correlation test was employed to assess the relationship between total antioxidant levels and various parameters. The level of statistical significance was set at < 0.05. Results The mean duration of tobacco use was 6.30 years in group 1 and 18.10 years in group 2 (p=0.001). The mean salivary flow rates were 0.272 ± 0.137 mL/min (group 1), 0.159 ± 0.094 mL/min (group 2), and 0.277 ± 0.192 mL/min (group 3), with a p-value of 0.02. The mean pH levels were 6.999 ± 0.322 (group 1), 6.650 ± 0.355 (group 2), and 6.943 ± 0.360 (group 3), with a value of 0.004. Group 3 had significantly higher total salivary antioxidant levels (=0.001). Conclusions Smokeless tobacco use, particularly in individuals with tobacco pouch keratosis, is associated with reduced salivary flow rate, lower (more acidic) pH, and decreased total antioxidant levels. There is a strong negative correlation between salivary antioxidant levels and the duration and frequency of smokeless tobacco use.