Only the dried stigma of the saffron, a flower deemed as the most valuable spice globally, is utilized for industrial production. Hence, there exists a growing interest in utilizing saffron floral bio‐residues. The anti‐hyperuricemic activity of a flavonoid extract from saffron floral bio‐residues was assessed in potassium oxonate‐induced hyperuricemia mice. In addition, an ultra‐high performance liquid chromatography‐triple quadrupole mass spectrometry method was established and validated to determine the pharmacokinetics of five main flavonoids and three phase‐II metabolites in rat plasma after oral administration of the flavonoid extract for the first time. Compared with pharmacokinetic parameters of kaempferol‐3‐O‐sophoroside, the most abundant flavonoid in the extract, and its aglycone kaempferol, we observed that coexisting compounds significantly reduced the absorption, accelerated the excretion of kaempferol‐3‐O‐sophoroside, while significantly increasing the absorption and prolonging the residence time of kaempferol in the flavonoid extract. These results suggest the promising potential of the flavonoid extract from saffron floral bio‐residues as an anti‐hyperuricemic agent. Kaempferol was absorbed in plasma at high concentrations owing to the biotransformation of kaempferol glycosides in vivo.
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