Background: Interleukin 8 (IL-8) or CX-CL8 is a powerful chemoattractant chemokine molecule and an inflammatory and angiogenesis-promoting cytokine, which acts as a biomarker in saliva in patients with tobacco users, oral cancers, and potentially malignant disorders. Aims and Objective: To evaluate and compare salivary IL-8 levels in patients with tobacco users, oral cancer, and potentially malignant disorders and healthy individuals using ELISA (enzyme-linked immunosorbent assay). Materials and Method: Eighty samples were grouped into four sub-groups, with 20 samples in each group: Group I: 20 oral cancer patients; Group II: 20 potentially malignant disorders; Group III: 20 tobacco users; Group IV: 20 control subjects. A sample of unstimulated saliva of 5 mL was collected from the selected individuals based on the exclusion and inclusion criteria. Saliva was centrifuged at 3500 revolutions per minute (rpm), and ELISA for IL-8 was carried out in saliva, stored at -4°C. Results: IBM Statistical Package for Social Sciences (SPSS) for Windows 11, Version 23.0 (Armonk, New York: IBM Corp) software was used for the analysis of data. Using one-way analysis of variance and Tukey's test for post-hoc analysis, the significant difference in the multi-variate analysis was determined. The mean value of salivary IL-8 levels in Groups I, II, II, and IV was 1055.7, 1074.0, 857.9, and 781.7, respectively. The P value between the groups is 0.77 (p > 0.05), which is statistically not significant. Conclusion: This study found that tobacco users have greater salivary IL-8 levels than individuals with potentially malignant diseases and oral malignancies and healthy patients. © 2023 Elsevier B.V., All rights reserved.