Abstract: OBJECTIVE To investigate the metabolic regularity of ampelopisn in rat liver microsomes and the effect of pharmaceutical excipients on its metabolism. METHODS The UPLC-MS/MS method was established to screen metabolic conditions in vitro and evaluate metabolic patterns by detecting the residual concentration of ampelopsin in the metabolic reaction system. RESULTS Ampelopisn metabolism was affected by incubation time, liver microsome concentration and initial ampelopisn concentration. Rat hepatocyte pigment P450 subenzyme CYP3A, CYP1A1/2 and CYP2E1 played a major role in serpentine metabolism. The inhibitory effect of pharmaceutical excipients on ampelopsin metabolism was dose-dependent manner. The metabolic kinetics studies revealed that Cremophor RH40, Tween 80, PVP K30, HPBCD and F68 exhibited significant inhibition metabolism in a mixed competition. CONCLUSION These pharmaceutical excipients are expected to improve the oral bioavailability of ampelopsin by inhibiting metabolism.