UHPLC-Q-Orbitrap HRMS分析吉马酮大鼠体内代谢产物及代谢途径

目的 研究吉马酮在大鼠体内的代谢产物，探讨其可能的代谢途径。方法 采用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱技术（UHPLC-Q-Orbitrap HRMS）对灌胃给予吉马酮（10 mg·kg^-1）后大鼠的生物样品（血浆、尿液和粪便）进行分析，色谱柱采用Waters ACQUITY UPLC^® BEH C₁₈（2.1 mm×50 mm，1.7 μm），流动相乙腈-0.1%甲酸水溶液，梯度洗脱，Q-Exactive高分辨质谱仪采用HESI离子源，辅助气温度300℃，离子传输管温度320℃，喷雾电压为+3.50 kV或者-2.80 kV，扫描方式Full MS/dd-MS²。分析比对给药组和空白组中高分辨质谱提供的准分子离子峰和碎片离子信息，使用Xcalibur 3.0、Mass Frontier 2.0和Compound Discovery 2.1软件进行数据处理，筛选并鉴定其可能的代谢产物。结果 从大鼠生物样品中共筛选鉴定了包括原型在内的23个代谢产物，推断吉马酮在大鼠体内发生的代谢类型主要有脱水反应、脱饱和反应、氧化反应、葡萄糖胺结合反应、葡萄糖醛酸化反应以及相关的复合反应等。结论 本研究较为详细地阐明了吉马酮在大鼠体内的代谢情况，剖析了其代谢途径，可为其进一步的药效学、药动学及毒理学研究提供理论依据。

Metabolites and Metabolic Pathway Analysis of Germacrone in Rats by UHPLC-Q-Orbitrap HRMS

OBJECTIVE To study the metabolites of germacrone in rats and explore its possible metabolic pathway. METHODS The biological samples of rats(plasma, urine and feces) were analyzed after intragastric administration germacrone(10 mg·kg^-1) by UHPLC-Q-Orbitrap HRMS. The chromatographic Waters ACQUITY UPLC^® BEH C₁₈(2.1 mm×50 mm, 1.7 μm) column was used and the mobile phase was acetonitrile-0.1% formic acid aqueous solution with gradient elution. The Q-Exactive high-resolution mass spectrometer equipped HESI ion source was adopted. Auxiliary temperature and ion transport tube temperature were set at 300℃ and 320℃. Spray voltage was +3.50 kV or -2.80 kV and scanning mode was Full MS/dd-MS². The quasi-molecular ion peaks and fragment ion information provided by high resolution mass spectrometry in administration group and blank group were analyzed and compared. Xcalibur 3.0, Mass Frontier 2.0 and Compound Discovery 2.1 software were used for data processing to screen and identify potential metabolites. RESULTS Twenty three protocompound and metabolites were screened and identified in the biological samples of rats, and the main metabolic reactions of germacrone in rats were dehydration reaction, desaturation reaction, oxidation reaction, glucosamine binding reaction, glucosaldehyde acidification reaction and related complex reaction, etc. CONCLUSION In this study, the metabolic process of germacrone in rats is elucidated in detail, and its metabolic pathway is analyzed, which can provide theoretical basis for the further study of pharmacodynamics, pharmacokinetics and toxicology of germacrone.