Phospholipid-linked glycation has been implicated to play an important role in lipid peroxidation associated with several conditions, such as normal aging and diabetes. In this study, we established a convenient method for the preparation of a high-purity, Amadori-glycated phosphatidylethanolamine (Amadori-PE) standard in a methanol system. We evaluated the prooxidant effect of the Amadori-PE standard. When Amadori-PE was incubated in a micellar and liposome system in the presence of metal ion, a remarkable increase of lipid peroxidation was observed in both systems. In addition, the Amadori-PE-induced lipid peroxidation was effectively inhibited by superoxide dismutase, mannitol, catalase, and EDTA. These results indicate that Amadori-PE is a considerable risk factor that may cause membrane lipid peroxidation in the pathogenesis of diabetes and aging.