蒺藜茎叶化学成分及含量测定研究

  本文关键词：蒺藜茎叶化学成分及含量测定研究，，由笔耕文化传播整理发布。

        蒺藜(Tribulus terrestris L.),为蒺藜科(Zygophylaceae)蒺藜属(Tribulus)植物。蒺藜茎叶又名蒺藜苗,始载于《本草纲目》,具有祛风、消肿作用,主治痈肿,疥癣,风痒,鼻塞等。蒺藜在我国具有悠久的药用历史,素有“草中名药”之称,其果实、花、根等也均可作为药用部位。蒺藜主要含有皂苷、黄酮、生物碱和多糖类等化合物,现代药理研究表明甾体皂苷和黄酮类成分为其主要活性成分,具有改善心脑血管、降血糖、增强性功能、抗衰老、调血脂、抑菌、抗肿癌等药理活性。本文首先通过查阅国内外大量文献,对蒺藜化学成分及药理作用研究现状进行了综述,共引用153篇文献。在此基础上,采用化学手段以及多种分离手段,包括萃取、大孔树脂、日本MCI树脂、羟丙基葡聚糖凝胶色(Sephadex LH20)及PHPLC等分离手段对蒺藜茎叶95%乙醇提取物部位进行了化学成分研究,共分离得到14个化合物,通过理化性质及UV、1hnmr、13cnmr、hmbc、hsqc等波谱技术鉴定出6个化合物,分别为异鼠李素-3-O-β-D-龙胆双糖苷(isorhamnetin-3-O-β-D-gentiobioside, JL-1),槲皮素-3-O-β-D-龙胆双糖苷(quercetin-3-O-β-D-gentiobioside, JL-2),山奈酚-3-O-β-D-龙胆双糖苷(kaemp ferol-3-O-β-D-gentiobioside, JL-3),异鼠李素-3-O-β-D-龙胆双糖基-7-O-β-D-吡喃葡萄糖苷(isorhamnetin-3-O-β-D-gentiobioside-7-0-P-D-glucoside, JL-4),山奈酚-3-O-β-D-龙胆双糖基-7-I-β-D-吡喃葡萄糖苷(kaempferol-3-O-β-D-gentiobioside-7-0-β-D-glucoside, JL-5), N-反式-对羟基苯乙基阿魏酰胺-4’-0-βD-葡萄糖苷(N-trans-ferulo yl tyramine-4’-O-β-D-glucopyranoside, JL-6)。化合物JL-5和JL-6中糖的连接方式首次明确,JL-6为首次从蒺藜科植物中分离得到。JL-1, JL-2及JL-4在蒺藜中含量较高,经检测纯度均达到99%以上,符合对照品的要求。另外,对蒺藜茎叶有效部位的质量控制方法进行了研究,采用HPLC-DAD法建立了蒺藜茎叶中含量较高的三种黄酮醇苷类化合物的含量测定方法。结果,异鼠李素-3-O-β-D-龙胆双糖基-7-O-β-D-葡萄糖苷、槲皮素-3-O-β-D-龙胆双糖苷及异鼠李素-3-O-β-D-龙胆双糖苷分别在0.56～14.00、3.24～129.60及0.92～23.00μg/mL范围内线性良好,其回归方程分别为Y=21388X-2087.8, r=0.9996; Y=34596X-37335, r=0.9998; Y=50667X+10439, r=0.9998。本文通过对蒺藜茎叶化学成分的研究,从分离出的黄酮化合物中,选择含量较高的三个黄酮醇类化合物作为对照品。利用反相HPLC法同时检测了异鼠李素-3-O-β-D-龙胆双糖基-7-O-β-D-葡萄糖苷、槲皮素-3-O-β-D-龙胆双糖苷及异鼠李素-3-O-βp-D-龙胆双糖苷的含量,该法具有简便、快速、稳定可靠、灵敏的特点,精密度及重现性良好,回收率符合要求,可为蒺藜或其制剂的质量控制和综合评价提供参考。

    The plant Tribulus terrestris L., belonging to the family Zygophylianceae, have been used in traditional Chinese medicine against various diseases. The stems and leaves of Tribulus terrestris L., also known as "Jili-miao", only contained in the "Shen Nong’s Herbal Classic" are used for treating edema, abdominal distension and so on. As we all know, besides the magical therapeutic effect of the fruit of Tribulus terrestris L., its roots, flowers, stems and leaves can also be used as medicine parts. Tribulus terrestris L. has contained numerous compounds such as saponins, flavonoids, alkaioids and polysaccharides. Modern pharmacological studies have shown that steroidal saponins and flavonoids are mainly active constituents, which can improve cardiovascular diseases, lowering blood sugar, enhancing sexual function, anti-aging and lipids, anti-neoplasm pharmaceutical activity.Firstly, the paper summarizes the research situation of chemical constituents and p harmacological activities on the basis of the current domestic and international literatur es (153references were cited). By means of macroporous resin, MCI gel, the Sephade x LH-20, preparative HPLC separation method et al.14compounds were isolated from the95%EtOH extract of stems and leaves. Among them, on the basis of spectroscop y analysis, including UV,1HNMR,13CNMR, HMBC, HSQC and chemical methods,5compound structures were identified as isorhamnetin-3-O-β-D-gentiobioside (JL-1), querc etin-3-O-β-D-gentiobioside (JL-2), kaempferol-3-O-β-D-gentiobioside (JL-3), isorhamnetin-3-O-β-D-gentiobioside-7-O-β-D-glucoside(JL-4), kaempferol-3-O-β-D-gentiobioside-7-O-β-D-glucoside(JL-5), N-trans-feruloyl tyramine-4’-O-β-D-glucopyranoside(JL-6). Among the m, compound JL-6was the first reported in Zygophylaceae. Compounds JL-1, JL-2an d JL-4with the purity of more than99%were chosen as the reference substance. Esta blish a quantitative method for the determination of three flavonols in the effective of the stems and leaves of Tribulus terrestris by HPLC-ADA. The contents of index com pounds including isorhamnetin-3-O-β-D-gentiobioside (JL-1), quercetin-3-O-β-D-gentiobio side (JL-2) and isorhamnetin-3-O-β-D-gentiobioside-7-O-β-D-glucoside(JL-4) were deter mined by HPLC. The calibration curve is linear at ranges of0.56-14.00,3.24～129.60and0.92～23.00μg/mL respectively. Their regression equations are Y=21388X-2087.8, r=0.9996; Y=34596X-37335, r=0.9998; Y=50667X+10439, r=0.9998。In this paper, by the isolation and structural identification of flavonoids compound from the stems and leaves of Tribulus terrestris L, we confirmed higher content of th ree flavonol compounds as reference substance. Establish a method based on reversed phase HPLC was developed to simultaneously determine the content of isorhamnetin-3- O-β-D-gentiobioside, quercetin-3-O-β-D-gentiobioside and isorhamnetin-3-O-β-D-gentiobio side-7-O-P-β-glucoside. This method is considered to be simple, accurate, stable, good precision and reproducibility and is suitable for evaluating the quality of Tribulus terre stris L.

蒺藜茎叶化学成分及含量测定研究

中文摘要4-6ABSTRACT6-7前言8-10Abbreviations10-12第一部分 文献综述12-30    第一节 概述12    第二节 蒺藜化学成分研究进展12-20    第三节 蒺藜药理作用研究进展20-30第二部分 蒺藜茎叶实验研究部分30-68    第一章 蒺藜茎叶化学成分研究30-46        第一节 化学成分结构解析30-38        第二节 实验部分38-46    第二章 蒺藜茎叶黄酮对照品研究46-58        第一节 对照品结构鉴定46-47        第二节 对照品纯度测定47-57        第三节 结果与讨论57-58    第三章 HPLC法测定蒺藜茎叶中3个黄酮苷的含量58-68总结与讨论68-70主要创新点70-72参考文献72-82附图82-102致谢102-104个人简历104

  本文关键词：蒺藜茎叶化学成分及含量测定研究，由笔耕文化传播整理发布。