Flavonol–serum albumin complexation. Two-electron oxidation of flavonols and their complexes with serum albumin

Abstract

Quercetin (3,3′,4′,5,7-pentahydroxyflavone) and quercetin derivatives (3-methylquercetin, isoquercitrin, rutin) are strong polyphenolic antioxidants abundant in plants and in the human diet. Recent investigations have shown that significant concentrations of albumin-bound quercetin conjugates are present in the plasma of humans fed a quercetin-rich diet.

In this work, binding of quercetin and quercetin glycosides to bovine serum albumin (BSA) is quantitatively investigated by fluorescence spectroscopy. The strong fluorescence enhancement of quercetin upon binding points to the fact that a significant fraction of quercetin adopts a pyrylium-like structure in the complex. On the other hand, the observation of a very efficient quenching of tryptophan fluorescence by quercetin is consistent with a binding occurring in the IIA domain.

Flavonoid-derived quinones may be formed upon quenching of reactive oxygen species by flavonoids (antioxidant activity). In this work, the quinones are conveniently formed upon periodate oxidation of the selected flavonoids in methanol and in aqueous buffers with and without BSA. A kinetic investigation by UV–visible spectroscopy shows that albumin-bound flavonoids are oxidized as quickly as free flavonoids. Interestingly, the quercetin quinone, which is merely detectable in the absence of BSA because of fast solvent addition, is efficiently stabilized in the complex by charge transfer interactions (pH 9). No evidence for quercetin–BSA conjugates could be found, thus showing that water addition (and subsequent degradation) remains the sole significant pathway of quinone transformation in the complex.