葛根芩连汤中葛根素、黄芩苷、小檗碱与甘草酸组合在大鼠体内代谢产物研究

Studies on Metabolites of Main Components of Gegen Qinlian Decoction in Rats

摘要:

目的: 研究葛根芩连汤中各味药材的主要成分葛根素、黄芩苷、小檗碱与甘草酸联合给药后在大鼠体内的代谢产物化学结构。 方法: 健康大鼠灌胃给药,收集0~24 h的尿样,尿样经C₁₈固相小柱分离纯化后,经UPLC-Q-TOF-MS分析鉴定其中的代谢产物。代谢物的结构鉴定主要依据各代谢物与对照品的一级质谱电离规律、裂解规律间的关联性和相关文献的数据。 结果: 灌胃给药(葛根素、小檗碱、黄芩苷和甘草酸)后,在大鼠尿液中鉴定了16个代谢产物的化学结构,其中6个来自葛根素,分别为葛根素、大豆苷、大豆苷元、二氢大豆苷元、葛根素硫酸酯结合产物和葛根素葡萄糖醛酸结合产物;2个来自黄芩苷,分别为baicalin-6-O-β-glucose-7-O-β-glucopyranuronoside,baicalin-6-O-β-glucopyranuronoside-7-O-β-glucopyranuronoside;8个来自小檗碱,分别为thalifendine,berberrubine,demethyleneberberine,jatrorrhizine,demethyleneberberine葡萄糖醛酸结合物,jatrorrhizine葡萄糖醛酸结合物,berberrubine-9-O-β-D-glucuronide,demethyleneberberine-2,3-O-β-D-glucuronide。 结论: 主要成分联合给药后,代谢产物主要以苷元或以葡萄糖苷、葡萄糖醛酸结合物为主,也有以原型排出;甘草酸生物利用度低,有明显的首过效应,且经尿液排泄量极少,未检出代谢产物。

Abstract:

Objective: To study the metabolism of combination adiminastration of main components of Gegen Qinlian decoction in rats. Method: The urine samples were collected 0-24 h after ig main components of Gegen Qinlian decoction,then the samples were purified through C₁₈ solid-phase extration cartridge. The purified samples were analyzed by UPLC-Q-TOF-MS. Identification and structural elucidation of the metabolites were performed by comparing the changes in molecular masses, retention times and full scan metabolisms(MS) spectra with those of the parent drug,and the relative data of authentic reference. Result: Sixteen metabolism were identified as puerarin,daidzin,daidzein,dihydrodaidzein, puerarin 4'-O-β-sulfate, puerarin-7-O-glucuronide, baicalin-6-O-β-glucose-7-O-β-glucopyranuronoside, baicalin-6-O-β-glucopyranuronoside-7-O-β-glucopyranuro-noside, thalifendine, berberrubine,demethyleneberberine,jatrorrhizine,demethyleneberberine-glucuronide,jatrorrhizine-glucuronide, berberrubine-9-O-β-D-glucuronide,demethyleneberberine-2,3-O-β-D-glucuronide. Conclusion: Oxidative demethylenation and subsequent glucuronidation are the major metabolic pathways of combination adiminastration of main components of Gegen Qinlian decoction, some components were mainly excreted unmetabolized in the urine; because of lower bioavailability,first-pass effect in intestine of glycyrrhizic acid,and only a minor part eliminated by urine,the metabolisms of it were not detected.