The present study aimed to investigate the human serum proteome wide interaction of aluminium oxide nanoparticles (Al2O3-NPs). Methodologically, the specific interaction of serum proteins with Al2O3-NPs were explored through high resolution proteomic profiling. Here we employed LC-MS/MS based HRMS protocol to explore the list of interacted proteins. Further, novel human serum proteins interacted with Al2O3-NPs and its associated molecular pathways have been illustrated.