Saliva is an accessible biofluid with growing diagnostic potential, yet the influence of collection methods on its proteome is not well understood. This study compared the proteomic profiles of stimulated and unstimulated whole human saliva. Saliva samples were collected from five healthy adult males under unstimulated conditions and following citric acid stimulation, then analyzed using 2D liquid chromatography–mass spectrometry. A total of 509 non-redundant proteins were identified, with 288 proteins compared across conditions. Thirty-six proteins showed significant changes in relative abundance after stimulation. These findings demonstrate that salivary stimulation alters the whole saliva proteome, with important implications for saliva-based biomarker discovery and diagnostics.
