大鼠灌胃缩泉方制剂后尿、粪中的益智活性成分及代谢物的测定

摘要

Objective To identify the flavonoids (tectochrysin, izalpinin, chrysin, kaempferide and apigen-in-4',7-dimethylether) and their metabolites, as well as diarylheptanoids (yakuchinone A, yakuchinone B and oxy-phyllacinol) and their metabolites, in rat urine and feces after oral administration of Suoquan capsules or pills. Methods After male Sprague Dawleyoral rats were administratered Suoquan pills (3.54 g/kg) and Suoquan capsules (3.77 g/kg), they were placed into the metabolism of the cage. Their urine and dung were collected 12 h later. The dung was weighed and added 9 volumes of deionized water and then homogenized at high speed divided method. 100μl homogenate of urine and dung were taken into centrifuge tube, with 300μl methanol added, shaked 5 min by vortex and centrifuged 10 min at 13 000 r/min. The supernatant were drawed and dried under N2. The residue was reconstitut-ed using 50μl methanol liquid, shaked and centrifugated again. 40μl supernatant was drawed and analyzed by liguid chromatography-tandem mass spectrometry (LC-MS/MS). Results According to the peak LC-MS/MS case in the bi-ological sample of the mentioned control group, comparing the urine and dungs sample before and after the adminis-trated Suoquan pills and Suoquan capsules, we found that the main substances detected in urine and dungs were proto-type form and were not detected the phaseⅡmetabolites, such as glucuronide metabolites, in rats after intragastric ad-ministration. Conclusion Nootkatone, puzzle ketone A and Young shoots flavin and other ingredients are detected in the urine of rats, which may be the active constituents in regulating the bladder and bladder function.%目的：研究大鼠灌胃缩泉胶囊和缩泉丸后，诺卡酮、益智黄酮(杨芽黄素、伊砂黄素、白杨素、山奈素和芹菜素-4'，7-二甲氧醚)和益智中二芳基庚烷类成分(益智酮甲、益智酮乙、益智醇)在尿和粪中的物质形式。方法雄性Sprague Dawley经灌胃给予缩泉胶囊(3.54 g/kg)和缩泉丸(3.77 g/kg)后置于代谢鼠笼中，收集12 h的尿液和粪球。粪球称重后加入9倍体积的去离子水，高速匀浆。分别取100μl尿液和粪球匀浆液于离心管中，加入300μl甲醇，漩涡振摇5 min，13000次/min下离心10 min，取上清液，在N2下吹干，残渣用50μl甲醇复溶，再次振摇和离心，取上清液40μl，利用液相色谱-串联质谱联用仪(LC-MS/MS)进行分析。结果根据上述对照品在生物样品中的LC-MS/MS出峰情况，通过比较空白尿、粪样品与给药后相应的尿粪样品，研究发现，大鼠灌胃后，在尿液和粪便中主要检测到原形物质形式，未检测到葡萄糖醛酸结合物等二相代谢产物。结论大鼠尿液中检测到的诺卡酮、益智酮甲以及杨芽黄素等成分，可能是直接作用于膀胱而发挥调节膀胱功能的活性成分。