The lack of diagnosis at its early and treatable stage is the major challenge associated with tobacco-associated oral diseases including oral cancer. Thus, it is highly desirable to delineate the molecular basis of tobacco-associated pathogenesis in the oral cavity to discover a sensitive, robust, and selective analytical tool for tobacco-related oral health. Mucins (MUC) are a family of high-molecular-weight O-linked glycoproteins, which act as the first barrier in case of any external insult to the oral cavity. To explore whether long-term habituation of high doses of tobacco influences oral MUC status, secretory MUC and its saccharides were determined from the saliva collected from bidi smokers, smokeless tobacco (gutkha and tobacco powder) habituates, and healthy, nonsmoking individuals. Interestingly, a notable reduction in the secretory MUC content was noticed in the saliva from tobacco cohorts. The duration of tobacco exposure has been identified as the main factor influencing oral mucin secretion. Specifically, the aberrant terminal glycosylation of MUC emerges as the promising diagnostic tool, to trace the molecular changes associated with tobacco use in the oral cavity.