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Serum albumin is the most abundant protein in the circulatory system. It plays an important role in the transport and deposition of many molecules in the body. The 7-O-β-d-glucopyranosyl-6-(3-methylbut-2-enyl)-5,4′-dihydroxyflavonol (PF) is a glycosylated flavonoid isolated from Ouratea hexasperma (Ochnaceae) branches. It shows important biological activities, such as cytotoxic, antimicrobial and antitumor. The aim of this study was to examine the interaction of bovine serum albumin (BSA) with PF in a PBS buffer solution (pH = 7.4) at 296 K, 303 K and 310 K, by spectroscopic techniques (UV–vis, circular dichroism, FTIR-ATR, steady-state, time-resolved, synchronous and 3D fluorescence) and molecular docking. The experimental results showed that the intrinsic fluorescence quenching of BSA induced by PF resulted in an association BSA:PF in the ground state (k_q 10¹² M⁻¹ s⁻¹). The association is moderate …