Background: With its complex pathogenesis and high mortality, acute lung injury (ALI) is closely associated with a poor prognosis in critically ill patients. The present study used network pharmacology to investigate the mass spectrometry (MS) behaviors and molecular mechanisms of Amadori compounds acting on ALI and diabetes.Methods: Two typical Amadori compounds (Fru-Trp and Glc(α1,4)-Fru-Trp) were used to study the fragmentation mechanisms by tandem MS in negative and positive-ion modes. The potential targets of Amadori compounds on ALI were screened from a public database. R package was used for the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and the protein-protein interactions (PPIs) of the targets were evaluated by Search Tool for the Retrieval of Interacting Genes (STRING). Finally, molecular docking was implemented to analyze the binding energies and action modes of the target compounds through the online tool CB-Dock.Results: In relation the common precursor ions of the Amadori compounds, the sodium adducting cations provided the most abundant fragmentation information in c for analyzing their chemical structures. 103 and 109 intersecting targets of glucose-Amadori and maltose-Amadori, respectively, were separately identified in the ALI database, and significant overlaps between the GO terms and KEGG pathways were noted. Three shared hub targets (i.e., vascular endothelial growth factor A, caspase-3, and proto-oncogene tyrosine-protein kinase) were selected from the 2 PPI networks by STRING. The molecular docking results showed that the Amadori compounds had good binding potential to the hub target proteins, and that Amadori compounds had lower binding energies and more stable conformations than their corresponding carbohydrates.Conclusions: As the endogenous compounds in diabetes, Amadori compounds may act on and activate a wide range of protein receptors, which may also produce bi-directional regulatory effects that influence ALI progression. Thus, the risk factors of diabetes in the progression of ALI should be carefully assessed, and the molecular mechanisms of Amadori compounds in animal models for ALI should be further verified.