梅晓丹;王喻淇;王子健;赵文靖;尚展鹏;乔延江;张加余;
MEI Xiao-dan;WANG Yu-qi;WANG Zi-jian;ZHAO Wen-jing;SHANG Zhan-peng;QIAO Yan-jiang;ZHANG Jia-yu;School of Chinese Pharmacy, Beijing University of Chinese Medicine;Beijing Institution of Chinese Medicine, Beijing University of Chinese Medicine;

• 1:北京中医药大学中药学院
• 2:北京中医药大学北京中医药研究院

摘要(Abstract)：

应用UHPLC-LTQ-Orbitrap技术对丹参素在大鼠血浆和尿液中的代谢产物进行鉴定。SD大鼠灌胃给药丹参素CMC-Na混悬液后,收集其血浆和尿液,应用固相萃取法进行样品前处理。采用ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱。在电喷雾负离子模式下采集生物样品的质谱数据。根据精确相对分子质量以及多级质谱裂解信息,结合对照品及文献报道结果比对,最终鉴定了丹参素以及21个丹参素Ⅰ相和Ⅱ相代谢产物,同时分析了丹参素主要的体内代谢反应,如脱水、甲基化、葡萄糖醛酸化、硫酸酯化、去羟基化以及相关的复合反应等。该文应用高分辨液质联用技术快速阐明了丹参素在大鼠体内的代谢产物,为今后药效物质基础和作用机制研究奠定了基础。
To identify the metabolites of Danshensu in plasma and urine in rats by using UHPLC-LTQ-Orbitrap method. After oral gavage of Danshensu CMC-Na suspension in SD rats, urine and plasma samples were collected and processed by solid phase extraction. ACQUITY UPLC BEH C_(18) column(2.1 mm ×100 mm, 1.7 μm) was utilized, with 0.1% formic acid(A)-acetonitrile(B) solution as the mobile phase for gradient elution. Negative electrospray ion mode based data-acquisition method was established to collect the mass spectrometry data of biological samples. As a result, Danshensu and 21 Danshensu Ⅰ phase and Ⅱ phase metabolites were finally identified according to the accurate mass measurements, mass fragmentation behaviors and comparing with the reference standards. The main metabolic pathways included dehydration, methylation, glucuronide conjugation, sulfate conjugation and their composite reactions. Consequently, our study expounded metabolites of Danshensu in rats based on UHPLC-LTQ-Orbitrap method and provided a reference for further researches on therapeutic material basis and mechanism of Danshensu.

关键词(KeyWords)： 丹参素; UHPLC-LTQ-Orbitrap;代谢产物;代谢途径
Danshensu;UHPLC-LTQ-Orbitrap;metabolites;metabolic pathways

Abstract:

Keywords:

基金项目(Foundation): 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);; 江苏省中药药效与安全性评价重点实验室项目(JKLPSE201816);; 2017年北京市科技新星与领军人才培养专项(Z171100001117029);; 国家自然科学基金项目(81430094)

作者(Authors): 梅晓丹;王喻淇;王子健;赵文靖;尚展鹏;乔延江;张加余;
MEI Xiao-dan;WANG Yu-qi;WANG Zi-jian;ZHAO Wen-jing;SHANG Zhan-peng;QIAO Yan-jiang;ZHANG Jia-yu;School of Chinese Pharmacy, Beijing University of Chinese Medicine;Beijing Institution of Chinese Medicine, Beijing University of Chinese Medicine;

DOI: 10.19540/j.cnki.cjcmm.20180709.010

参考文献(References)：

• [1] 中国药典. 一部[S]. 2015.
• [2] 刘艾林,李铭源,王一涛,等.丹参药理学活性物质基础研究现状[J].中国药学杂志,2007(9):641.
• [3] 胡天鑫,文爱东,朱艳荣,等. 丹参素、羟基红花黄色素A单用及合用对大鼠心肌缺血/再灌注损伤的保护作用[J].中国现代中药,2015,17(1):15.
• [4] 白清.丹皮酚联合丹参素对糖尿病大鼠的内皮功能及血液流变学的影响[J].中国实验方剂学杂志,2015,21(3):110.
• [5] 封亮,张明华,顾俊菲,等.基于组分结构理论的丹参滴注液的多维结构过程动态质量控制研究[J].中国中药杂志,2013,38(21):3622.
• [6] 俞翔.丹参提取和醇沉过程的质量控制技术研究[D].杭州:浙江大学, 2008.
• [7] 贾海红.丹七片等中药复方制剂的质量控制方法与研究[D].石家庄:河北医科大学, 2015.
• [8] Wang X, Li W, Ma X, et al. Identification of a major metabolite of Danshensu in rat urine and simultaneous determination of Danshensu and its metabolite in plasma: application to a pharmacokinetic study in rats [J]. Drug Test Anal, 2015, 7(8): 727.
• [9] Gu J F, Feng L, Zhang M H, et al. New metabolite profiles of Danshensu in rats by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry [J]. J Chromatogra B, 2014, 955: 20.
• [10] Zhang J Y, Wang Z J, Zhang Q, et al. Rapid screening and identification of target constituents using full scan-parent ions list-dynamic exclusion acquisition coupled to diagnostic product ions analysis on a hybrid LTQ-Orbitrap mass spectrometer [J]. Talanta, 2014, 124: 111.
• [11] Zhang J Y, Wang Z J, Li Y, et al. A strategy for comprehensive identification of sequential constituents using ultra-high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometer, application study on chlorogenic acids in Flos Lonicerae Japonicae [J]. Talanta, 2016,147(62):16.
• [12] Zhang J Y, Cai W, Zhou Y, et al. Profiling and identification of the metabolites of baicalin and study on their tissue distribution in rats by ultra-high-performance liquid chromatography with linear ion trap-Orbitrap mass spectrometer [J]. J Chromatogr B, 2015, 985: 91.
• [13] 刘思燚,张秀平,尚展鹏,等. UHPLC-LTQ-Orbitrap鉴定苦碟子注射液化学成分及其在大鼠血浆中的移行成分[J].中国中药杂志,2016,41(12):2235.
• [14] 王子健,刘颖,刘思燚,等.UPLC-HRMSn结合高能诱导裂解快速鉴定麦冬中高异黄酮类成分[J].质谱学报,2016,37(6):481.
• [15] 尚展鹏,王菲,王子健,等.UHPLC-LTQ-Orbitrap分析硫磺熏蒸麦冬中高异黄酮硫酸酯和亚硫酸酯[J].质谱学报,2018,39(2):180.
• [16] Shang Z P, Wang F, Dai S Y, et al. Profiling and identification of (-)-epicatechin metabolites in rats using ultra-high performance liquid chromatography coupled with linear trap-orbitrap mass spectrometer[J]. Drug Test Anal, 2017,9(8):1224.
• [17] 赵文靖,黄漠然,尚展鹏,等.丹参酮Ⅰ和丹参酮ⅡA在大鼠体内的代谢物研究[J].中国中药杂志,2018,43(1):174.
• [18] 蔡伟.苦碟子倍半萜内酯类似物靶向分离及体内代谢研究[D].北京:北京中医药大学,2015.
• [19] Zhang Z C, Xu M, Sun S F, et al. Metabolic analysis of four phenolic acids in rat by liquid chromatography-tandem mass spectrometry[J]. J Chromatogr B, 2008,871(1):7.
• [20] 魏华.创新药物丹参素钠的体内分析及其代谢研究[D].上海:第二军医大学,2010.