摘要
目的:研究大鼠分别灌胃和静脉给药葫芦茶苷溶液后,血浆中的葫芦茶苷及其代谢产物。方法:雌性Sprague Dawley大鼠分别经灌胃(10mg/kg)和静脉注射(2mg/kg)给予葫芦茶苷溶液后,按设计的时间点从大鼠眼內眦静脉丛采集血液样品,置于肝素化的离心管中,低温离心得血浆。取5、15、30min和1、2h血浆样品各40μL,合并,混匀,加入600μL甲醇,漩涡振摇5min,13 000r/min离心10min,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。根据葫芦茶苷在血浆样品中的LC-MS/MS出峰情况,比较空白血浆样品与不同途径给药后相应的生物样品,结果:大鼠血浆中可以检测到葫芦茶苷原形物质,也可检测到脱糖代谢物对羟基桂皮酸。通过中性丢失扫描,确定对羟基桂皮酸再发生结合代谢形成硫酸酯化代谢物。同时,对羟基桂皮酸发生脱氢反应,形成对羟基苯丙酸。结论:给药后大鼠血浆中形成葫芦茶苷多种代谢物,对羟基桂皮酸可能是发挥药效活性的代谢产物。
Objective： To identify tadehaginoside and its metabolites in rat plasma after oral （p. o. ） or intravenous （i. v. ） administration of tadehaginoside solution to rats. Methods： The Sprague Dawley rats were randomly divided into two groups of three rats each to identify the metabolites of tadehaginoside after a single intravenous （i. v. ）dose （2 mg/kg） or per oral （p. o. ） dose （10 μ/kg） of tadehaginoside solution. Serial blood samples were collected and heparinized for plasma prepara- tion. For identification of tadehaginoside and its metabolites, the collected plasma samples at different time points （5, 15, 30 min and 1, 2 h） were pooled for each group and aliquots （200 μL each） were treated with 600 μL methanol and then mixed by vortex-shaking for 10 min and centrifuged at 18,140 g for 10 min. The upper supernatant was dried under a stream of N2. The residue was reconstituted in 50 μL of methanol, centrifuged ditto, and 10μL of the resulting supernatant were applied to LC-MS/MS analysis. The tadehaginoside and its metaholites were identified using liquid chromatography/tandem mass spectrome- try （LC-MS/MS） with selected reaction monitoring mode. Results： Compared with the LC-MS/MS spectrum of standard chemicals, tadehaginoside, p-coumaric acid and phloretic acid in rat plasma were identified. Meanwhile, the p- coumaric acid was sulfated to conjugated metabolite. Conclusion： Tadehaginoside and three metabolites were identified in rat plasma using LC-MS/MS. The p-eoumaric acid might be the active metabolite of tadehaginoside. Further research is needed to evaluate the activities of tadehaginoside and p-coumaric acid.
出处
《海南医学院学报》
CAS
2015年第2期145-147,151,共4页
Journal of Hainan Medical University
基金
国家自然科学基金资助项目(81202994
81460629)~~
