绿原酸在肝微粒体和肠道菌群中的体外代谢研究

In vitro research on metabolism of chlorogenic acid in liver microsomes and intestinal flora

中文关键词:  绿原酸  肝微粒体  肠道菌群  代谢途径  超高效液相色谱-四极杆飞行时间质谱法

英文关键词:chlorogenic acid  liver microsome  intestinal flora  metabolic pathway  ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry

中文摘要:

      目的 研究绿原酸在大鼠肝微粒体和肠道菌群中的代谢产物及代谢途径。方法 采用超高效液相色谱-串联四极杆飞行时间质谱法(ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry, UPLC-Q-TOF/MS)系统分析了肝微粒体和肠道细菌试液中绿原酸及其代谢物。色谱柱为Waters ACQUITY UPLC BEH C18 (2.1 mm×100 mm, 1.7 μm), 流动相为乙腈和0.1% (V/V)甲酸水溶液梯度洗脱系统, 流速为0.35 mL/min, 离子源为电喷雾离子源(electrospray ion source, ESI), 以负离子模式进行检测。采用Masslynx软件通过保留时间与一、二级质谱信息, 进行成分分析。结果 大鼠肝微粒体温孵样品中有绿原酸原型成分和奎宁酸、咖啡酸2种代谢产物; 粪便温孵样品中有绿原酸原型成分和奎宁酸、3-羟基苯丙酸、二氢咖啡酸、二氢阿魏酸、咖啡酸、O-甲基绿原酸6种代谢产物。结论 UPLC-Q-TOF/MS技术鉴定绿原酸及其代谢产物, 操作便捷高效, 结果准确全面, 绿原酸在体外代谢的主要途径为氢化、甲基化、水解、去羟基化等。

英文摘要:

      Objective To study the metabolic pathway of chlorogenic acid in rat liver microsomes and intestinal flora by component analysis of chlorogenic acid and its metabolites. Methods Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) with negative electrospray ion source (ESI-) was applied to systemically analyze chlorogenic acid and its metabolite in the liver microsomes and intestinal bacteria test solution. The conditions were as follows: The chromatographic column was Waters ACQUITY UPLC BEH C18 (2.1 mm×100 mm, 1.7 μm), the mobile phase was acetonitrile-0.1% formic acid (V/V) aqueous solution (gradient elution), the flow rate was 0.35 mL/min. Masslynx software was used to analyze the components by retention time and primary and secondary mass spectrometry. Results Chlorogenic acid and 2 kinds of metabolites (quinic acid, caffeic acid) were detected in the incubated samples of liver microparticles. Chlorogenic acid and 6 kinds of metabolites (quinic acid, 3-hydroxyphenylpropionic acid, dihydrocaffeic acid, dihydroferulic acid, caffeic acid, and O-methyl- chlorogenic acid) were detected in fecal incubation samples. Conclusion UPLC-Q-TOF/MS is a convenient and efficient method for the identification of chlorogenic acid and its metabolites. The main metabolic pathways of chlorogenic acid in vitro are hydrogenation, methylation, hydrolysis and dehydroxylation, et c.