Abstract: It is of great significance to search for advanced glycation end products (AGEs) inhibitors with high activity and low side effects from food-borne natural products for the prevention and improvement of related diseases. To elucidate the inhibitory effect of tectoridin on the generation of advanced glycation end products (AGEs), the glycation system of bovine serum protein (BSA)-methylglyoxal (MGO) was established, the effects of tectoridin on the AGEs inhibition, glycation degree, protein oxidation products, protein cross-linking and MGO trapping were investigated by spectroscopy, SDS-PAGE, HPLC, LTQ-Orbitrap MS, and molecular docking. The results showed that tectoridin increased the content of lysine and thiol, reduced the contents of protein carbonylation and protein oxidation products, alleviated the cross-linking structure of BSA. The mono-MGO adduct of tectoridin was identified in the incubation of MGO with tectoridin. Molecular docking results showed that hydrogen bonding and hydrophobic interactions were the major driving forces for stabilizing tectoridin-BSA complexes. Tectoridin was a promising natural AGEs inhibitor. Mechanism analysis revealed that tectoridin alleviated AGEs formation by trapping MGO to form mono-tectoridin-MGO complexes and forming stable tectoridin-BSA complexes. These findings would provide the theoretical basis for the development of tectoridin as a novel, natural and effective inhibitor of AGEs.