[Material basis of Wuzhuyu Decoction in treatment of chronic migraine by UPLC-Q-Exactive-MS/MS]

The classical prescription Wuzhuyu Decoction(WZYD) has demonstrated distinctive efficacy in alleviating migraine and improving gastrointestinal symptoms. However, its pharmacodynamic material basis remains insufficiently elucidated. In this study, a chronic migraine rat model was established using inflammatory soup induction. Following successful model establishment and observed therapeutic efficacy of WZYD, samples were collected from the WZYD treatment group after the final administration. These included drug-containing plasma, cerebrospinal fluid, bile, urine, feces, as well as in vitro incubation samples of WZYD with liver homogenate and fecal microbiota. Ultra-high performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry(UPLC-Q-Exactive-MS/MS) was employed to comprehensively analyze the in vivo and in vitro metabolic profiles of WZYD. A total of 169 compounds were identified or tentatively characterized, comprising 46 prototype compounds and 123 metabolites. Specifically, 18 prototype and 48 metabolized alkaloids, 6 prototype and 20 metabolized limonoids, 7 prototype and 18 metabolized ginsenosides, 3 prototype and 17 metabolized gingerols, 5 prototype and 11 metabolized flavonoids, and 7 prototype and 9 metabolized organic acids were detected. In summary, this study provided a systematic characterization of the in vivo and in vitro metabolic constituents of WZYD in the context of chronic migraine treatment, thereby elucidating its pharmacodynamic material basis and offering valuable insight for the further development and utilization of this classical prescription.