目的：研究大鼠灌胃缩泉胶囊后益智中二芳基庚烷类成分(益智酮甲、益智酮乙、益智醇)在大鼠胆汁中的物质形式。方法雄性Sprague Dawley大鼠胆管插管手术完毕经灌胃给予缩泉胶囊(1.64 g/kg)后收集0~4 h胆汁。同体积(100μl)的两份胆汁样品分别经蛋白沉淀和β-葡萄糖醛酸水解酶(1000 U/ml，pH为5的乙酸钠缓冲液)处理后，所得样品利用液相色谱-质谱联用仪(LC-MS/MS)进行分析。结果在大鼠胆汁中几乎检测不到益智二芳基庚烷类成分的原形物质形式，而是以葡萄糖醛酸结合物的形式存在。这些二相代谢物经酶水解处理后代谢物的色谱峰消失，释放出二芳基庚烷类成分的原形物质。结论大鼠灌胃给药后经肠道吸收的二芳基庚烷类成分在胆汁中形成葡萄糖醛酸结合产物，酶水解可以释放出原形物质形式。本研究可以帮助我们理解这类物质的体内转运过程，也为后期的定量分析工作奠定基础。%Objective To identify the diarylheptanoids (yakuchinone A, yakuchinone B and oxyphyllacinol) and their metabolites in rat bile after oral administration of Suoquan capsules. Methods Cannulas were surgically in-serted into the bile duct of male Sprague-Dawley rats (n=3) under anesthesia (pentobarbital, 50 mg/kg, i.p.) and bile was collected for 4 h after oral administration of Suoquan capsules (1.64 g/kg, suspended in 0.5%w/v sodium carboxy-methycellulose). After methanol precipitation or β-glucuronidase (1 000 units/ml in pH 5 acetate buffer) treatment, these chemicals and their phase Ⅱ metabolites in rat bile samples were analyzed via liquid chromatography/tandem mass spectrometry (LC-MS/MS) with selected reaction monitoring mode. Results The parent compounds could not be measured in rat bile sample treated by methanol precipitation. On the contrast, glucuronidated metabolites were recovered in rat bile. After enzymatic hydrolysis treatment viaβ-glucuronidase, the corresponding parent chemicals were librated from their conjugated forms. Conclusion After oral administration, the absorbed diarylheptanoids were partly biotransformed to glucuronide conjugates and excreted into rat bile. This study is helpful for better un-derstanding of these diarylheptanoids’fates in rat and quantitative analysis of these compounds in rat bile samples in the future.
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