Quercitrin, one of quercetin glycosides, has been demonstrated to be a powerful antioxidant in vitro. However, its bioavailability is not well defined. In the present study, we investigated the absorption and metabolism of quercitrin on a Caco-2 cell model. The concentrations of quercitrin, quercetin, isorhamnetin and tamarixetin in Caco-2 cells were quantified using a HPLC/MS method after incubation with quercitrin. Meanwhile, other possible metabolites of quercitrin were also detected. The results showed that quercitrin, quercetin, isorhamnetin and tamarixetin were measurable in Caco-2 cells after incubation with quercitrin. Their intracellular concentrations rose up to the peak value 60 min post incubation, and then decreased to the plateau gradually. Furthermore, quercetin monoglucuronide, quercetin trisulphate and qurecetin glucuronide sulfate were identified in the Caco-2 cells treated with quercitrin. It is concluded that quercitrin can be uptaken intact by Caco-2 cells and subsequently undergoes metabolic transformations, including methylation, glucuronidation and sulfation.