中国中药杂志

2020, v.45(16) 3952-3960

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鼠尾草酸在大鼠体内代谢产物的UHPLC-Q-Exactive MS鉴定研究
Analysis of carnosic acid metabolites in rats by UHPLC-Q-Exactive MS

娄天宇;马贝贝;梁耀月;王晨晓;刘金辉;李瑞吉;于尚玥;王志斌;
LOU Tian-yu;MA Bei-bei;LIANG Yao-yue;WANG Chen-xiao;LIU Jin-hui;LI Rui-ji;YU Shang-yue;WANG Zhi-bin;School of Chinese Pharmacy, Beijing University of Chinese Medicine;Beijing Institution of Tongrentang;

摘要(Abstract):

采用UHPLC-Q-Exactive MS对鼠尾草酸在大鼠体内的代谢产物进行分析鉴定。大鼠灌胃鼠尾草酸后,收集其血浆、尿液、粪便等生物样品,利用固相萃取法对生物样品进行前处理。采用Acquity UPLC BEH C_(18)色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸溶液(A)-乙腈(B)为流动相进行梯度洗脱。在正、负离子模式下对生物样品进行分析。根据精确相对分子质量、碎片离子信息及相关文献报道,共筛选和鉴定了28种鼠尾草酸的代谢产物。该研究发现,鼠尾草酸在大鼠体内的代谢途径主要为氧化、羟基化、甲基化、葡萄糖醛酸化、硫酸酯化、S-半胱氨酸结合、谷胱甘肽结合、脱甲基、脱羰基以及它们的复合反应等。应用UHPLC-Q-Exactive MS高效全面地阐明了鼠尾草酸在大鼠体内的代谢情况,可为明确其药效物质基础和代谢机制提供依据。
A method of ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field Obitrap high-resolution mass spectrometry(UHPLC-Q-Exactive MS) was established to comprehensively identify the metabolites of carnosic acid in rats. After oral gavage of carnosic acid CMC-Na suspension in rats, urine, plasma and feces samples were collected and pretreated by solid phase extraction(SPE). Acquity UPLC BEH C_(18 )column(2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid solution(A)-acetonitrile(B) as the mobile phase for the gradient elution. Biological samples were analyzed by quadrupole/electrostatic field Obitrap high-resolution mass spectrometry in positive and negative ion mode. Based on the accurate molecular mass, fragment ion information, and related literature reports, a total of 28 compounds(including carnosic acid) were finally identified in rat samples. As a result, the main metabolic pathways of carnosic acid in rats are oxidation, hydroxylation, methylation, glucuronide conjugation, sulfate conjugation, S-cysteine conjugation, glutathione conjugation, demethylation, decarbonylation and their composite reactions. The study showed that the metabolism of carnosic acid in rats could be efficiently and comprehensively clarified by using UHPLC-Q-Exactive MS, providing a reference for clarifying the material basis and metabolic mechanism of carnosic acid.

关键词(KeyWords): 鼠尾草酸;UHPLC-Q-Exactive MS;代谢产物;代谢途径
carnosic acid;UHPLC-Q-Exactive MS;metabolites;metabolic pathways

Abstract:

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基金项目(Foundation): 国家药典委员会项目(12);; 江苏省中药药效与安全性评价重点实验室项目(JKLPSE201816);; 中医脑病学山西省重点实验室(山西中医药大学)项目(CME-OP-2017002)

作者(Author): 娄天宇;马贝贝;梁耀月;王晨晓;刘金辉;李瑞吉;于尚玥;王志斌;
LOU Tian-yu;MA Bei-bei;LIANG Yao-yue;WANG Chen-xiao;LIU Jin-hui;LI Rui-ji;YU Shang-yue;WANG Zhi-bin;School of Chinese Pharmacy, Beijing University of Chinese Medicine;Beijing Institution of Tongrentang;

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