【作者】 王璐；

【导师】 姜宏梁；

【作者基本信息】 华中科技大学 ， 药物分析学， 2019， 硕士

【摘要】 中药泽泻(Alismatis Rhizoma)作为一味历史悠久的中药,在传统中医中常用于治疗肾炎水肿、小便不利、痰饮晕眩等症。在现代药理学研究中发现,泽泻中的三萜类成分具有明显的降血脂、抗动脉粥样硬化等作用。已有的安全性评价研究也已经证明泽泻肝肾毒性较小,适合于长期用药。然而,关于其体内代谢的研究还未见报道。因此,本课题运用超高效液相色谱串联质谱技术,基于泽泻三萜类化合物特征离子数据库,开发了一个快速、准确的鉴定体内代谢物的分析策略,对口服泽泻三萜标准品和泽泻提取物的大鼠主要进行了体内代谢研究,对泽泻三萜类成分的体内代谢产物进行了全面的表征。方法:1.泽泻三萜标准品在大鼠体内的代谢研究根据泽泻三萜类化合物在质谱裂解过程中的产生的特征离子,可将其分为三类,具有特征离子m/z383.3和339.3的I型泽泻三萜、具有特征离子m/z353.3的II型泽泻三萜及具有特征离子m/z381.3和339.3的III型泽泻三萜。选取alisol B-23acetate,alisol C-23 acetate及alisol F作为对应的代表性标准品进行体内代谢研究。基于UHPLC-QTOF-MS/MS技术,以0.1%甲酸水溶液和乙腈为洗脱溶剂,使用Ultimate-XB C18 column色谱柱及0.3m L/min的流速,在正离子模式下对口服泽泻标准品的大鼠血浆、尿液、粪便及胆汁样品进行全扫描并采集数据。2.泽泻提取物在大鼠体内的代谢研究基于泽泻三萜标准品的体内代谢研究基础,采用相同的分析方法对泽泻提取物的体内代谢过程进行研究。结果:1.在泽泻三萜标准品的体内代谢研究中,标准品alisol B-23 acetate在体内共发现了30种成分,包括原型化合物以及29个代谢产物;标准品alisol C-23 acetate在体内发现了原型化合物以及24种代谢产物;标准品alisol F在体内也发现了原型化合物及26种代谢物。2.对于泽泻提取物在大鼠体内的代谢研究,在血浆、尿液、粪便及胆汁样品中共鉴定出233种成分,其中包括原型化合物85种,代谢物148种。胆汁样品中检测到的代谢产物最多,有112个,血浆样品中最少,仅有28个。3.泽泻三萜类化合物的体内代谢过程中,加氢、氧化、硫酸化和葡萄糖醛酸结合是主要代谢途径。结论:本研究以泽泻三萜代表性的标准品和泽泻提取物为研究对象,运用液质联用技术分别对单一化合物和复杂中药成分的体内代谢产物进行了研究。本研究首次揭示了泽泻在体内的代谢产物及代谢过程,不仅为泽泻生物活性成分的研究提供了科学依据,而且对泽泻的药效学和药代动力学研究具有一定的参考价值。更多还原

【Abstract】 Alismatis Rhizoma(AR)is a traditional Chinese medicine with a long history and has been used for the treatment of nephritis edema,dysuria,and dizziness.In modern pharmacological study,it was found that the triterpenes in AR can lower serum lipids and reduce atherosclerosis.AR can be used for a long time because existing safety evaluation studies have shown that it has minimal kidney and liver toxicity.However,there are few reports about the metabolism of AR in vivo.Therefore,we developed an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method,which was based on the characteristic ions of established database of knwon triterpenes.It was used to analyze the major metabolites in rat after oral administration of three representative standards and Alismatis Rhizoma extracts(ARE).Method:1.Study on metabolism of three triterpenes standards of AR in vivoThe triterpenes of AR can be divided into three types according to their characteristic ions.Type I tritepenes represented by alisol B-23 acetate with characteristic ions of m/z 383.3 and 339.3,type II tritepenes represented by alisol C-23 acetate with characteristic ions of m/z 353.3 and type III tritepenes represented by alisol F with characteristic ions of m/z 381.3 and 339.3,respectively.Firstly,the plasma,bile,urine and feces samples of rats were collected after oral administration of the standards of triterpenes.Then,based on UHPLC-QTOF-MS/MS,the chromatography separation was performed on an Ultimate-XB C18 column with the mobile phase of 0.1% formic acid aqueous solution and acetonitrile,and the flow rate was 0.3 m L/min.Data acquisition was performed in positive ionization mode in mass spectrometry analysis.2.Study on metabolism of ARE in vivoBased on the in vivo metabolism of triterpene standards of AR,the in vivo metabolism of ARE was studied using the same method.Result:1.For the in vivo metabolism study of triterpene standards in AR,30 components including prototype compounds and 29 metabolites has been found for alisol B-23 acetate.24 metabolites except prototype compounds was found for alisol C-23 acetate.26 metabolites were found for alisol F except prototype.2.For the in vivo metabolism study of ARE,233 components,including 85 prototype compounds and 148 metabolites,were identified in plasma,urine,feces and bile samples.Most of metabolites were identified in bile samples,ang only 28 metabolites were identified in the plasma samples.3.For the in vivo metabolism study of triterpenes in AR,the major metabolic pathways include dehydrogenation,hydrogenation,oxidation,deacaylation,sulfation,glucuronidation and combined multiple-step metabolism.Conclusion:The metabolic profile of triterpenes from Alismatic Rhizoma in vivo was characterized by UHPLC-QTOF-MS/MS in this study.It is the first time to study the in vivo metabolism of AR,which not only provides a scientific basis for the study of the bioactive components,but also provides valuable information for the continuous study of the pharmacodynamics and pharmacokinetics.更多还原