[Identification of icaritin metabolites in rat plasma by UPLC-QTOF-MS]

To identify metabolites of icaritin in rat plasma by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS).

Acetonitrile was employed to precipitate protein in rat plasma. The separation was carried out on a BEH C18 column (2.1 mm x 50 mm,1.7 microm) using a gradient mobile phase of acetonitrile, water and ammonium formate. High resolution mass spectrometry was applied for qualitative analysis in positive and negative ion modes, respectively. The metabolites were predicted by the PALLAS software and identified in the UNIFI database.

Twenty-four metabolites of icaritin in rat plasma which were generated by reactions of hydroxylation, hydrogenation, epoxidation, acetylation, demethylation, dehydroxylation, sulfation conjugation, glucuronidation and glycosylation, were identified.

A rapid and efficient UPLC-QTOF-MS method is established for the identification of icaritin metabolites in rat plasma, which provided support for the preclinical safety evaluation of icaritin.