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基于超高效液相色谱-线性离子阱-静电场轨道阱质谱技术对大鼠体内丹酚酸A与丹酚酸B代谢过程的分析与比较

[Analysis and comparison of metabolic processes of salvianolic acid A and salvianolic acid B in rats based on UHPLC-LTQ-Orbitrap MS technology].

作者信息

Ma Bei-Bei, Lou Tian-Yu, Liang Yao-Yue, Wang Ting-Ting, Li Rui-Ji, Liu Jin-Hui, Wang Chen-Xiao, Yu Shang-Yue, Guo Yu-Dong, Wang Jing, Wang Zhi-Bin

机构信息

School of Chinese Pharmacy, Beijing University of Chinese Medicine Beijing 102488, China.

Beijing Institute for Drug Control Beijing 102200, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2021 May;46(9):2276-2286. doi: 10.19540/j.cnki.cjcmm.20200820.201.

Abstract

The metabolites of salvianolic acid A and salvianolic acid B in rats were analyzed and compared by ultra-high-perfor-mance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS). After the rats were administrated by gavage, plasma at different time points and urine within 24 hours were collected to be treated by solid phase extraction(SPE), then they were gradient eluted by Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) and 0.1% formic acid solution(A)-acetonitrile(B) mobile phase system, and finally all biological samples of rats were analyzed under negative ion scanning mode. By obtaining the accurate relative molecular mass and multi-level mass spectrometry information of metabolites, combined with the characteristic cleavage law of the reference standard and literature reports, a total of 30 metabolites, including salvianolic acid A and B, were identified. Among them, there were 24 metabolites derived from salvianolic acid A, with the main metabolic pathways including ester bond cleavage, dehydroxylation, decarboxylation, hydrogenation, methylation, hydroxylation, sulfonation, glucuronidation, and their multiple reactions. There were 15 metabolites of salvianolic acid B, and the main biotransformation pathways were five-membered ring cracking, ester bond cleavage, decarboxylation, dehydroxylation, hydrogenation, methylation, sulfonation, glucuronidation, and their compound reactions. In this study, the cross-metabolic profile of salvianolic acid A and B was elucidated completely, which would provide reference for further studies on the basis of pharmacodynamic substances and the exploration of pharmacological mechanism.

摘要

采用超高效液相色谱-线性离子阱-轨道阱质谱联用技术(UHPLC-LTQ-Orbitrap MS)对大鼠体内丹酚酸A和丹酚酸B的代谢产物进行分析比较。大鼠灌胃给药后,收集不同时间点的血浆及24小时内的尿液,经固相萃取(SPE)处理,然后采用Acquity UPLC BEH C₁₈柱(2.1 mm×100 mm,1.7 μm)和0.1%甲酸溶液(A)-乙腈(B)流动相体系进行梯度洗脱,最后在负离子扫描模式下对大鼠所有生物样品进行分析。通过获取代谢产物的精确相对分子质量和多级质谱信息,结合对照品的特征裂解规律及文献报道,共鉴定出包括丹酚酸A和B在内的30种代谢产物。其中,丹酚酸A的代谢产物有24种,主要代谢途径包括酯键裂解、脱羟基、脱羧、氢化、甲基化、羟基化、磺化、葡萄糖醛酸化及其多重反应。丹酚酸B的代谢产物有15种,主要生物转化途径为五元环开裂、酯键裂解、脱羧、脱羟基、氢化、甲基化、磺化、葡萄糖醛酸化及其复合反应。本研究全面阐明了丹酚酸A和B的交叉代谢谱,为进一步开展药效物质基础及药理机制探索研究提供参考。

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