桂枝茯苓胶囊在原发性痛经模型大鼠血浆、胆汁、尿液和粪便中的代谢产物研究

目的:研究桂枝茯苓胶囊在原发性痛经模型大鼠体内的代谢产物。方法:基于超高效液相色谱-线性离子阱-静电轨道阱串联质谱联用(UHPLC-LTQ-Orbitrap-MS)方法,采用Waters ACQUITY UPLC BEH C₁₈色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-0.1%甲酸水为流动相,梯度洗脱,流速0.4 mL·min^-1,质谱采用电喷雾(ESI)离子源,以正负离子模式采集多级质谱碎片信息,对大鼠灌服桂枝茯苓胶囊混悬液后的血浆、胆汁、尿液和粪便进行分析,利用质量亏损过滤(MDF)技术进行代谢产物鉴定分析。结果:共鉴定出8个原形及50个代谢产物,在体内主要发生Ⅰ相代谢反应(脱羧、水解、脱羟基和脱水等)及Ⅱ相代谢反应(甲基化、乙酰化、硫酸化及葡萄糖醛酸化等)。结论:采用LC-MS方法鉴定了原发性痛经模型大鼠体内桂枝茯苓胶囊代谢产物,为该复方制剂的药物代谢动力学研究提供依据。

Analysis of metabolites in primary dysmenorrhea rats plasma,bile,urine and feces after administration of Guizhi Fuling capsules

Objective:To determine the metabolites in primary dysmenorrhea rats after administration of Guizhi Fuling capsules.Methods:The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric(UHPLC-LTQ-Orbitrap-MS)and mass defect filter techniques with electrospray ion(ESI)source were applied to collect multi-level mass spectrometry fragment information in both positive and negative ion modes.The separation was performed on a Waters ACQUITY UPLC BEH C18 column(100 mm×2.1 mm,1.7μm)with mobile phase of acetonitrile-water(containing 0.1%formic acid)by gradient elution at a flow rate of 0.4 mL·min^-1.After the administration of Guizhi Fuling capsules,the plasma,bile,urine and feces in rats were analyzed,and the metabolites were identified by mass loss filtration(MDF).Results:8 prototypes and 50 metabolites were determined.The in vivo main metabolic reactions were phaseⅠreaction(decarboxylation,hydrolysis,dehydroxylation and dehydration)and phaseⅡreaction(methylation,acetylation,sulfonation and glucuronidation).Conclusion:LC-MS isused to identify metabolites of Guizhi Fuling capsules in rats with primary dysmenorrhea,and this study provids a basis for pharmacokinetic studies of the compound preparation of traditional Chinese medicine.