Scope To develop a physiologically based kinetic (PBK) model that describes the absorption, distribution, metabolism, and excretion of hesperidin in humans, enabling the translation of in vitro concentration–response curves to in vivo dose–response curves. Methods and results The PBK model for hesperidin in humans was developed based on in vitro metabolic parameters. Hesperidin was predicted to mainly occur in the systemic circulation as different monoglucuronides. The plasma concentrations of hesperidin aglycone (hesperetin) was predicted to be <0.02 mg/L at an oral dose of 50 mg/kg bw. The developed PBK model allowed conversion of in vitro concentration–response curves for different effects to in vivo dose–response curves. The BMD05 (benchmark dose for 5% response) values for protein kinase A inhibition ranged between 135 and 529 mg/kg bw hesperidin, and for inhibition of endothelial cell migration and prostaglandin E2 and nitric oxide production ranged between 2.19 and 44 mg/kg bw hesperidin. These values are in line with reported human data showing in vivo effects by hesperidin and show that these effects may occur at Western dietary and supplementary intake of hesperidin. Conclusions The developed PBK model adequately predicts absorption, distribution, metabolism, and excretion of hesperidin in humans and allows to evaluate the human in vivo situation without the need for human intervention studies. Hesperetin and its metabolites have been reported to exert antiatherogenic and antiinflammatory activities based on in vitro data. The developed PBK model was used to facilitate the translation of these in vitro data to in vivo effective dose levels in humans. The values presented herein are in line with reported human oral dose levels showing in vivo effects by hesperidin and show that these effects may occur at Western dietary and supplementary intake of hesperidin.