Quercetin is one of the most commonly occurring flavonoids in nature. Although, quercetin and its metabolites express negligible fluorescence, the albumin-bound form of quercetin has a strong fluorescence property. Considering the structural variance of different albumins, we hypothesized that the fluorescence of albumin complexes of quercetin and its metabolites may vary significantly. Therefore, in this study the fluorescence enhancement of quercetin and some of its major metabolites in the presence of bovine (BSA), human (HSA), porcine (PSA), and rat serum albumins (RSA) were investigated by steady-state fluorescence spectroscopy in PBS buffer (pH 7.4). Among the tested quercetin metabolites, significant fluorescence signal was shown by albumin complexes of quercetin, isorhamnetin, and quercetin-3’-sulfate, while other metabolites (tamarixetin, quercetin-3-glucuronide, and isorhamnetin-3-glucuronide) expressed negligible fluorescence. BSA was the most potent enhancer of quercetin-3’-sulfate but it showed poor effects regarding other flavonoids. The strongest enhancement of isorhamnetin was caused by HSA, while it was less effective enhancer of quercetin and quercetin-3’-sulfate. PSA showed a strong fluorescence enhancement of quercetin and quercetin-3’-sulfate but it was poorly effective regarding isorhamnetin. RSA was the most potent enhancer of quercetin but it caused only a weak enhancement of isorhamnetin and quercetin-3’-sulfate. Large changes of the pH (such as pH 5.0 and pH 10.0) almost completely abolished the fluorescence signals of the complexes. Nevertheless, slight decrease (pH 7.0) reduced and slight increase (pH 7.8) generally enhanced the fluorescence of flavonoid-albumin complexes (only exceptions were quercetin-PSA and quercetin-RSA). Complex formations were also investigated by fluorescence quenching studies. Based on our results, the formations of quercetin-BSA, quercetin-HSA, isorhamnetin-BSA, isorhamnetin-HSA, isorhamnetin-PSA, and quercetin-3’-sulfate – HSA complexes followed 1:1 stoichiometry, while the presence of a secondary binding site of flavonoids was assumed regarding other tested albumin complexes. Our study highlights that albumins can induce significantly different fluorescence enhancement of flavonoids, and even the stoichiometry of flavonoid-albumin complexes may differ.
- fluorescence spectroscopy