BACKGROUND: Salivary proteomics technology can be used to evaluate the disease progression of Oral Submucous Fibrosis (OSMF), a morbid, crippling and premalignant condition of the oral mucosa, associated with the use of areca nut in various forms. AIMS: The objective of this preliminary study was to compare the proteomic profile of whole unstimulated saliva of subjects with OSMF with that of healthy volunteers to identify proteins, the levels of which were significantly altered between the two groups. SETTINGS AND DESIGN: Whole unstimulated saliva was obtained from 40 subjects with OSMF and age–sex matched healthy subjects METHODS AND MATERIALS: salivary proteins were separated using two–dimensional gel electrophoresis. Proteins, the levels of which were significantly different between the two groups, were identified by computer image analyses and subsequent MALDI–TOF–Mass Spectrometry. RESULTS: 184 protein spots were detected over control gel, while 169 on experimental gel. Significant no of protein spots were altered in both the groups, proteins which were found to be significantly different were immunoglobulin heavy chain variable region and methyl transferase. CONCLUSION: Comparison of the proteomic profile of whole unstimulated saliva of OSMF subjects with that of healthy control subjects revealed differential protein expression. The approach applied herein might be helpful in molecular insights of OSMF.