UGT-mediated metabolism plays a dominant role in the pharmacokinetic behavior and the disposition of morusin in vivo and in vitro.

•Developed a sensitive and reliable LC–MS/MS method to determine the pharmacokinetic behaviors of morusin and its four metabolites (three UGT conjugates and one CYP oxidative product) in rat plasma.•The absorption and disposition characteristics of morusin in rat intestine and bile in situ demonstrated the enterohepatic recycling may be the main reason for the double peaks of its pharmacokinetic curve.•UGT-mediated glucuronidation plays a dominant role in morusin metabolism using human recombinant CYPs, UGTs, as well as liver and intestinal microsomes.