基于UHPLC-Q-Orbitrap HRMS技术的补骨脂素在大鼠体内代谢产物的鉴定及代谢途径的分析

Metabolites and metabolic pathway analysis of psoralen in rats based on UHPLC-Q-Orbitrap HRMS

目的采用UHPLC-Q-Orbitrap HRMS技术对补骨脂素在大鼠体内的代谢产物进行鉴定分析。方法将SD大鼠单次灌胃给予补骨脂素羧甲基纤维素钠混悬液后,收集其不同时间点的血浆以及24 h内的尿液、粪便。样品前处理后,采用Waters Acquity UPLCBEH C18色谱柱(100 mm×2.1 mm,1.7μm);以体积分数0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱。在电喷雾离子源(ESI源)正、负离子模式下分析生物样本。结果基于精确分子质量、碎片离子信息及对照品的特征性裂解规律,共分析鉴定出15个代谢产物。结果表明水合、还原、氧化及谷氨酰胺结合是补骨脂素在大鼠体内的主要代谢途径。结论通过液质联用技术探究了补骨脂素在大鼠体内的代谢产物及其代谢途径,为其药效学及毒理学研究提供了重要的理论依据,同时也为药物代谢鉴定提供一种综合研究方法。

Objective To study the bioactive components of psoralen in vivo,the metabolites of psoralen in plasma,urine and feces in rats were probed using UHPLC-Q-Orbitrap HRMS.Methods After SD rats were administrated with a single dose of psoralen CMC-Na suspension by gavage,the plasma,urine and feces samples were collected at different time points within 24 hours.After being pretreated,these samples were separated on a Waters Acquity UPLCBEH C18 column(100 mm×2.1 mm,1.7μm)by gradient elution with 0.1%formic acid solution(A)-acetonitrile(B)as the mobile phase.Furthermore,Q-Orbitrap HRMS under positive MS/dd-MS2 mode was applied to analyze the components in these samples.Results A total of 15 metabolites were identified based on the precise molecular weight,fragment ion information and the characteristics of fragmentation pattern of the reference substance.Itriguingly,our results also showed that reduction,oxidation and glutamine conjugation were the main metabolic pathways of psoralen in rats.Conclusion This study reveals 15 metabolites of psoralen and the related metabolic pathways in rats,which provides an important theoretical basis for further research on pharmacodynamics and toxicology.Meanwhile our study can offer a comprehensive research method for drug metabolism identification.