Abstract: Methyl syringate, as a characteristic marker of specific honey varieties, can bind to human serum albumin (HSA) after entering the human body and achieve systemic transport through blood circulation, but their interaction mechanism still lacks systematic research. To address this, the methods mentioned by the title were employed to systematically investigate the fluorescence quenching characteristics, types of intermolecular forces, energy transfer mechanisms, and specific binding patterns between methyl syringate and HSA. As found by the results of fluorescence spectrometry, ultraviolet-visible absorption spectrometry and thermodynamic analysis, methyl syringate was bound to HSA with interaction distance of 5.82 nm through hydrogen bonds, van der Waals forces, and hydrophobic interaction forces, triggering non-radiative energy transfer that led to dynamic fluorescence quenching of HSA accompanied by a red shift in the maximum emission wavelength. As shown by the results of synchronous fluorescence spectrometry analysis, as the concentration of methyl syringate increased, the characteristic fluorescence intensities of tryptophan (Trp) residue and tyrosine (Tyr) residue gradually decreased, indicating decreased hydrophobicity decrease in the HSA microenvironment, the increase of peptide chain extension, and conformational changes in the protein. As demonstrated by the molecular docking model, methyl syringate could enter the binding pocket of HSA and form hydrogen bonds with amino acid residues, such as Tyr residue Tyr-138 and histidine (His) residue His-146, and this specific binding pattern might serve as the structural basis for the aforementioned phenomena. The dynamic interaction mechanism between methyl syringate and HSA was elucidated in this study, which could provide an important theoretical foundation for deeper understanding of the in vivo transport processes and potential physiological functions of honey´s characteristic components.