BACKGROUND Recently, it has been an increasing concern on the bioactivation and adverse reactions associated with consumption of herbal and nature products. 7-Ethoxycoumarin is one of coumarin family compounds, but little information is available regarding its potential reactive metabolites. METHOD 7-ethoxylcoumarin was incubated individually with human, monkey, dog and rat hepatocytes for 2 hr, metabolites were detected, identified and characterized using high resolution liquid chromagraphy - tandem mass spectrometry. RESULTS Twenty-eight metabolites (M1 - M28) were detected and identified. O-deethylation, glucuronidation, sulfation, oxygenation, oxidative ring-opening, hydrogenation, glutathionation, dehydrogenation, cysteination, glucosidation, methylation, and hydrolysis were observed. At least sixteen metabolites not reported previously, were newly identified. M1 (O-deethylation, mono-oxygenation and glucuronidation), M3 (O-deethylation and glucuronidation), M5 (hydrolysis and mono-oxygenation), M14 (O-deethylation), M16 (hydrolysis), M22 (oxidative ring-opening and oxygenation) and M27 (monooxygenation) exhibited high mass spectrometric responses in human hepatocytes. M3, M5, M8, M13 (mono-oxygenation), M14, M16, M18 (O-deethylation and sulfation), M22 and M27 exhibited high mass spectrometric responses in monkey hepatocytes. M14, M16, M18, M20 (glutathionation and dehydrogenation) and M27 exhibited high mass spectrometric responses in dog hepatocytes. M1 (Odeethylation, mono-oxygenation and glucuronidation), M3, M5, M13, M14, M16, M17 (cysteination), M18, M20, and M22 exhibited high mass spectrometric responses in rat hepatocytes. CONCLUSION Most of new metabolites via oxidative ring-opening and glutathionation were identified. Species differences in metabolism of 7-ethoxylcoumarin in hepatocytes were observed. The analysis of metabolites suggests that 7-ethoxylcoumarin may undergo 3,4-epoxidation responsible for formation of glutathione and its derived cysteine conjugates, carboxylic acid and its glucuronides, glucosides and sulfate.

中文翻译：

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高分辨率LC-MS / MS对人，猴，狗和大鼠肝细胞中7-乙氧基香豆素的体外药物代谢研究

背景技术近来，人们越来越关注与食用草药和天然产物有关的生物活化和不良反应。7-乙氧基香豆素是香豆素家族化合物之一，但关于其潜在的反应性代谢产物的信息很少。方法将7-乙氧基香豆素与人，猴，狗和大鼠的肝细胞分别孵育2小时，并使用高分辨率液相色谱-串联质谱法检测，鉴定和鉴定代谢产物。结果检测并鉴定了28种代谢物（M1-M28）。观察到O-脱乙基，葡糖醛酸化，硫酸化，氧合，氧化性开环，氢化，谷胱甘肽化，脱氢，半胱氨酸化，葡萄糖苷化，甲基化和水解。至少十六种代谢物以前未报道过，被新发现。M1（O-脱乙基，单加氧和葡糖醛酸化），M3（O-脱乙基和葡糖醛酸化），M5（水解和单加氧），M14（O-脱乙基），M16（水解），M22（氧化性开环和氧合）和M27（单氧合）在人肝细胞中表现出高质谱响应。M3，M5，M8，M13（单加氧），M14，M16，M18（O-脱乙基和硫酸化），M22和M27在猴肝细胞中表现出较高的质谱响应。M14，M16，M18，M20（谷胱甘肽化和脱氢）和M27在犬肝细胞中表现出高质谱响应。M1（去乙基化，单加氧和葡萄糖醛酸化），M3，M5，M13，M14，M16，M17（半胱氨酸化），M18，M20和M22在大鼠肝细胞中表现出高质谱响应。结论鉴定了大多数通过氧化开环和谷胱甘肽氧化作用的新代谢产物。观察到肝细胞中7-乙氧基香豆素代谢的物种差异。代谢物的分析表明7-乙氧基香豆素可能会经历3,4-环氧化作用，从而导致谷胱甘肽及其衍生的半胱氨酸结合物，羧酸及其葡糖醛酸，葡萄糖苷和硫酸盐的形成。