| Summary: | Ilaprazole is a proton pump inhibitor used to treat digestive diseases. In this study, blood samples were collected after oral administration of ilaprazole and prepared by liquid–liquid extraction. The metabolites of ilaprazole were detected by liquid chromatography–high-resolution mass spectrometry (LC-HRMS) and LC-MS<sup>n</sup>. A total of twelve in vivo metabolites were detected in rat plasma and six new metabolites of ilaprazole, including one reductive metabolite with sulfide (<b>M3</b>), two hydroxylated metabolites with sulfoxide (<b>M7</b> and <b>M8</b>), and three oxidative metabolites with sulfone (<b>M9</b>, <b>M11</b>, and <b>M12</b>), were identified. The possible metabolic pathways of ilaprazole and the fragmentation behaviors of its metabolites were elucidated. The result of the in silico prediction indicates that all the new metabolites showed the potential ability to inhibit H<sup>+</sup>/K<sup>+</sup>-ATPase activity.
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