中草药 2023年9月 第54卷 第17期

Abstract: Objective  To identify the metabolites of 5,7-dimethoxycoumarin and 6,7-dimethoxycoumarin in plasma, tissues (heart,  liver, spleen, lung, kidney, stomach and small intestine), urine and feces in rats by using hybrid quadrupole-orbitrap high resolution  mass spectrometry (UHPLC-Q-Orbitrap HRMS). Methods  SD male rats were ig 5,7-dimethoxycoumarin and 6,7 dimethoxycoumarin (100 mg/kg), the plasma, tissue, urine, and fecal samples were collected. After sample pretreatment, a Thermo  Scientific AccucoreT M C18 column (100 mm × 3 mm, 2.6 μm) was used, the gradient elution was performed with 0.1% formic acid- 0.1% formic acid acetonitrileas the mobile phase. The mass spectrometry data of biological samples were acquired in positive ion mode  with an electrospray ionization source (ESI source). Xcalibur was used to collect data, and the compound metabolite prediction template  of Compound Discoverer 3.1 software was used for analysis, and the metabolites were identified in combination with relevant literature  and Mass Frontier 7.0 software. Results  A total of 33 5,7-dimethoxycoumarin metabolites and 21 6,7-dimethoxycoumarin  metabolites were detected in biological samples from rats. A total of 8, 28, 23, 2, 10, 3, 10, 13, 7, 7 metabolites of 5,7 dimethoxycoumarin and 7, 19, 14, 7, 7, 4, 7, 13, 3, 8 metabolites of 6,7-dimethoxycoumarin were identified in plasma, urine, feces,  heart, liver, spleen, lung, kidney, stomach and small intestine, respectively. Conclusion  A total of 33 metabolites of 5,7 dimethoxycoumarin and 21 metabolites of 6,7-dimethoxycoumarin were identified by liquid chromatography-mass spectrometry. The metabolism of the two coumarins in rats was elucidated through demethylation, hydroxylation, hydrolysis, reduction, methylation,  sulfation and glucuronidation. The metabolic characteristics of the two coumarins were compared and analyzed. This study laid a  foundation for further study on the mechanism and pathway of action of the two coumarin in traditional Chinese medicine, and also  provided a comprehensive research method for drug metabolism identification. 
Key words: UHPLC-Q-Orbitrap HRMS; 5,7-dimethoxycoumarin; 6,7-dimethoxycoumarin; metabolites; metabolic pathway

本研究中对照品：5,7-二甲氧基香豆素（批号 1211325801）、6,7-二甲氧基香豆素（批号 1211201901）均购自于四川恒诚致远生物科技有限公司（Naturewill biotechnology Co., Ltd.）