HPLC-QTrap-MS鉴定欧前胡素体内代谢物及肝微粒体孵化产物

Identification of Imperatorin and Its Metabolites in Vitro and in Vivo by HPLC-QTrap-MS Technology

目的采用液质联用技术鉴定大鼠灌胃欧前胡素后尿液、粪便、胆汁及肝微粒体孵化液中的代谢产物,并探讨体内代谢与肝微粒体转化的相关性。方法联合应用多离子监测-信息依赖-增强型子离子扫描(MRM/MIM-IDA-EPI)、母离子扫描-增强分辨率扫描-信息依赖-增强型子离子扫描(PREC-ER-IDA-EPI)和增强型子离子扫描(EPI)多种模式,依据PREC-ER-IDAEPI中的[M+H]+和[M+NH4]+离子判断代谢物的相对分子质量,结合3种模式的EPI信息对代谢物进行结构表征。结果共鉴定出尿样中32个代谢物,粪样中14个代谢物,胆汁中6个代谢物以及肝微粒体转化17个代谢物。结论欧前胡素主要由肝脏代谢并经肾脏排泄。体外肝微粒体孵化结果与体内代谢比较表明,欧前胡素体内外代谢具有较好的相关性。

OBJECTIVE To identify the metabolites of imperatorin in rat urine, feces and bile after oral administration as well as the transformation products of imperatorin after incubation with rat liver microsomes with HPLC-QTrap-MS technology. METHODS The combination use of HPLC-QTrap-MS scanning mode including muhiple ion monitoring-information dependent acquisition-enhanced product ion （MRM/MIM-IDA-EPI）mode, precursor scan-enhanced resolution-information dependent acquisition-enhanced product ion （PREC-ER-IDA-EPI） mode and enhanced product ion （EPI） mode were performed for the identification of the metabolites. Based on the simultaneous appearance of [ M ＋ H ] ＋ and [ M ＋ NH4 ] ~ in the spectrum of PREC, the molecular weight could be unambiguously identified. The structures of compounds were then identified by the fragment ions generated from these three modes. RESULTS With the HPLC-QTrap-MS method, 32 metabolites in urine sample, 14 metabolites in faces sample, 6 metabolites in bile sample and 17 transformation products from the rat liver microsomes sample were detected. CONCLUSION Imperatorin is metabolized mainly in liver and excreted through kidney. The metabolic profiles of imperatorin in vivo and in vitro have good correlation.