Objective To explore the transmembrane absorption mechanism of quercetin based on sodium-dependent glucose transporter 1 (SGLT1) and glucose transporter 2 (GLUT2) in human intestinal epithelial cells (Caco-2) model. Methods Quercetin and buspirone (internal standard) were assessed by using the liquid chromatography-mass spectrometry (LC/MS) method. Transmembrane transport of quercetin was studied in a Caco-2 cell monolayer model, and the roles of SGLT1 and GLUT2 in this process were studied by using phloridzin (a potent inhibitor of SGLT1) and phloretin (a potent inhibitor of GLUT2). Quercetin was detected in receiver solution of the Caco-2 cell monolayer within 30~150 min. Results During 30~150min’s incubation, quercetin aglycon was detected among every group on the receiver side of transmembrane transport. The amount of quercetin in receiver solution increased at first and peaked at 120 min, and then decreased. Phloridzin exerted an obvious inhibitory effect on the transmembrane transport of quercetin, and meanwhile the phloretin significantly raised the amount of quercetin in the receiver side. Conclusions Quercetin was transported in a complete molecular form across the Caco-2 cell monolayer, and SGLT1 and GLUT2 might both be involved in its transmembrane transport simultaneously, but further research is needed to confirm the role of GLUT2 in the transmembrane transport of quercetin.