当归、川芎和归芎药对中主要芳香酸类成分在血虚大鼠体内的代谢研究

Identification on the metabolites of main aromatic acids from Danggui,Chuanxiong and the Gui-Xiong herb pair in blood deficiency rats

目的:研究当归、川芎和当归-川芎药对(归芎药对)中阿魏酸、绿原酸和咖啡酸在血虚大鼠血浆、尿液和胆汁中的代谢产物。方法:采用乙酰苯肼和环磷酰胺联合复制大鼠血虚模型,血虚大鼠灌胃当归、川芎和归芎药对提取物后,采用UPLC-Q-TOF/MS联用技术,利用碰撞能量梯度(MSE)和质量亏损过滤(MDF)技术,对其血浆、尿液和胆汁中的代谢产物进行分析。结果:根据离子碎片和代谢途径,当归、川芎和归芎药对提取物中阿魏酸、绿原酸和咖啡酸在血虚大鼠血浆、尿液和胆汁中推测出原型、还原、水解、甲基化、羟基化、硫酸酯化、葡萄糖醛酸化、磺酸化、甘氨酸结合、乙酰化、半胱氨酸结合和谷胱甘肽结合等共25个代谢产物。结论:当归、川芎和归芎药对提取物在血虚大鼠体内存在多种Ⅰ相和Ⅱ相代谢产物,为阐明该药对及其组成药味在体内发挥疗效的功效物质基础和作用机制提供依据。

Objectively： To study the major metabolites of ferulic acid （FA）, chlorogenic acid （ChA） and caffeic acid （CA） in Danggui （DG）, Chuanxiong （CX） and the Gui-Xiong herb pair （GX） extracts in blood deficient rats plasma, urine and bile. Methods： The blood deficiency model of rats was reproduced by phenylhydrazine and cyclophosphamide. Bit-samples were analyzed and detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS）. The metabolites of FA, ChA and CA in blood deficient rats plasma, urine and bile were identified after oral administration of DG, CX and GX extracts using MSE and mass defect filter techniques. Results： On the basis of the fragment ion and metabolic pattern, 25 metabolites including parent, reduction, hydrolysis, methylation hydroxylation, sulfate conjugation, gucuronide conjugation, taurine conjugation, glycine conjugation, acetylation, cysteine conjugation, glutathione conjugation and other metabolites of FA, ChA and CA were tentatively identified. Conclusion： The major metabolite of phase I and phase II of FA, ChA and CA in blood deficient rats plasma, urine and bile from DG, CX and GX extracts can be identified and provided referential information for material foundation of medicinal effectiveness and action mechanism of DG, CX and GX.