In this study, we implemented a highly robust workflow for O-phosphopeptides enrichment utilizing titanium-based immobilized metal affinity chromatography (Ti-IMAC). By integrating an offline reversed-phase prefractionation strategy and a combination of Electron-Transfer/Higher-Energy Collision Dissociation (EThcD) and Higher-Energy Collision Dissociation (HCD) mass spectrometry fragmentation techniques, we established the most comprehensive phosphoproteomic dataset for this species to our knowledge.