节点文献
中药大蓟中生物碱及多糖成分研究
Studies on Alkaloids and Polysaccharides from Cirsium Japonicum DC.
【作者】 黄新炜;
【导师】 段玉峰;
【作者基本信息】 陕西师范大学 , 有机化学, 2003, 硕士
【摘要】 大蓟为菊科(Compositae)管状花亚科蓟属(Cirsium Mill.)植物C.japonicum DC.的干燥地上部分或根,为我国传统中药,具有凉血止血、散瘀消肿之功效。 大蓟的化学成分复杂,具有广泛的药理作用和临床应用。现代化学和药理研究已部分阐明了其化学成分和临床应用之间的联系。例如,大蓟中的柳穿鱼叶苷为止血的有效成分;长链炔烯醇具有抗肿瘤作用,可作为治疗肝癌的依据。但是还有很多活性成分如抗菌、降压等成分未被找到,很多药理研究也仅停留在提取物阶段,因此对其化学成分和药理作用进行深入研究,进一步阐明其化学成分和临床应用之间的联系,对于指导临床用药和新药开发是非常必要的。目前,有关大蓟中生物碱及多糖的研究未见报道,因此本论文针对大蓟中的生物碱及多糖成分进行了研究。 本文第一部分为综述,首先是生物碱,包括生物碱的分类与分布、生物碱在植物化学分类上的意义、生物碱的生物活性、生物碱含量的测定方法、生物碱人工合成与结构的研究、细胞培养技术在生物碱生产中的应用以及生物碱的应用前景;其次,详细地论述了大蓟化学成分的研究现状、药理作用以及临床应用。 本文第二部分对大蓟中生物碱的提取、分离以及结构鉴定进行了研究。首先,通过生物碱与多糖的预试验,肯定了大蓟中既含生物碱,又含多糖。通过对大蓟中水溶性生物碱的检测,表明大蓟中不含水溶性生物碱,其中所含生物碱是亲脂性的,这样在进行生物碱提取时,可以不用考虑水溶性生物碱的影响。为了能够得到更多的总生物碱,便于进一步分离,,我们对生物碱三种不同提取方法,即水或酸水提取法、醇类溶剂提取法和脂溶性溶剂提取法的提取率进行了比较,其中以醇类溶剂提取法提取率最高,因此,我们选用95%乙醇作溶剂进行提取,醇提液浓缩所得浸膏用稀酸溶解,酸水液分别用氯仿、乙醚脱脂,氨水碱化,有机溶剂萃取,浓缩得总生物碱粗品。然后采用酸溶、碱化、有机溶剂萃取的方法对所得粗品进行了精制。采用重量法确定了大蓟中总碱含量的质量分数为0.02%。为了探求总碱中各单体完全分离的柱层析的最佳条件,以达到获得多个单体的目的,我们对总碱进行了薄层层析试验。根据薄层层析试验的结果,对总碱进行硅胶柱层析分离,分别用苯:乙酸乙酯:二乙胺=7:2:1和乙酸乙酯:氯仿:二乙胺=8:1:1进行洗脱,共分得两种生物碱单体CJ-Ⅰ和CJ-Ⅱ,用化学和光谱方法推断出CJ-Ⅰ可 能为去氢飞廉碱,CJ-11由于量太少,未能进行全面的结构鉴定。 本文第三部分对大蓟多糖的提取、纯化分离以及各级分分子量的测定进行了 研究。采用多糖传统的提取方法,即热水回流提取,水液浓缩后加乙醇析出沉淀, 再用95%乙醇和丙酮洗涤沉淀,真空干燥得多糖粗品KJ卜水提多糖中常含有蛋 白质及无机盐等大量小分子物质,这增加了多糖分离的难度,必须除去。我们采 用经典的Sevag法对粗糖进行了脱蛋白处理,用透析法除去小分子物质,效果良 好。经过脱蛋白和透析处理的多糖半纯品仍是混合物,需进一步按分子量的大小 分级。我们采用DEAE-纤维素柱层析法对多糖半纯品进行了分级,分别用水、 0.SMNaCI洗脱,分得水洗级分KJl)和盐洗级分oJ上),水洗级分较盐洗级分多。 采用重量法确定了样品中多糖的含量为3.6%,水洗级分的含量为59.4%,盐洗级 分的含量为 18.6%。最后采用凝胶过滤法测定了 CJ-l 和 CJ-2的分子量分别为 2石叫04,二24X105。 本文第四部分采用H。O。/Fe》体系法和APTEMED体系法对大蓟多糖进行了 抗氧化活性的研究,比较了CJ、CJI、CJ-2对OZ‘和·OH的清除率,得出CJ、 CJl、CJ毛对’OH都有一定的清除作用,且随着多糖浓度的增大,清除率升高。 其中CJ-1 的清除作用最强,在本实验所选取的浓度范围内,最高清除率可达 50.73%,CJ的清除作用较弱,而CJ-2的清除率随糖浓度的变化不是很明显;CJ 和 CJ刁对O。有一定的清除作用,且随着多糖浓度的增大,清除率升高。其中 CJl 的清除作用最强,在本实验所选取的浓度范围内,最高清除率可达67.35%,CJ 的清除作用较弱,而CJ-2对OZ’几乎没有清除作用。在进行抗氧化活性研究的同 时,采用滤纸片怯分别对CJ、CJ-1、CJ-2进行了体外抑菌活性的初步研究,所选 菌株为大肠杆菌和金黄色葡萄球菌。结果表明,CJ、CJq、CJ<分别显示程度不。等的抑菌作用。
【Abstract】 Cirsium japonicum DC., belonging to the Compositae family, is a common-used Chinese herbal drug. Having the efficacy of cooling the blood, hemostasis and detumescence, it has been mainly used to treat haematuria and carbuncle.Constituents of Cirsium japonicum DC. are complicated, and it has lots of pharmacological actions and clinical practices. Although modern chemical and pharmacological researches have partly elucidated the relation between chemical constituents and clinical practices, yet many active constituents, such as antibiosis and decompression, have not been found. Many pharmacological researches are only in use of the extract. So making a thorough research on its chemical constituents and pharmacological actions, elucidating the relation between chemical constituents and clinical practices further, are very necessary to guide clinical druging and develop new drugs. At present, there are no reports about alkaloids and polysaccharides from Cirsium japonicum DC.. So we studied alkaloids and polysaccharides from Cirsium japonicum DC..The first part of this thesis is a summary of alkaloids, including the classification and distribution of alkaloids, the significance of alkaloids in phytochemical classification, the bioactivities of alkaloids, the assaying methods of alkaloid’s content, the research on artificial synthesis and structure of alkaloids, the application of cell culture technology in alkaloid’s production and the prospect of alkaloid’s application. Next we discussed the present situation of chemical constituents, pharmacological actions and clinical practices of Cirsium japonicum DC..In the second part, we studied the extraction, isolation and structure identification of alkaloids. Above all, we confirmed that Cirsium japonicum DC. contained alkaloids and polysaccharides by pretest. Through the detection of water soluble alkaloids, we knew that alkaloids from Cirsium japonicum DC. is not water soluble, but liposoluble. So during the extraction, we needn’t consider the influence of water soluble alkaloids. In order to get more total alkaloids for isolation further, we compared the extraction rate of three different methods-water or acid water extraction, alcoholic solventextraction and liposoluble solvent extraction. The rate of alcoholic solvent extraction is the highest, so we selected 95%EtOH as solvent to extract. The alcoholic extract concentrated under reduced pressure, dissolved by diluted acid, degreased with CHC13 and Et2O, alkalified with NH3·H2O, extracted by organic solvent. At last we got coarse alkaloids after concentration. Then we refined it through acid dissolution alkalization and extraction by organic solvent. Next we determined the content of total alkaloids is 0.02% by gravimetric analysis. In order to search for the optimum isolated conditions of every monomer, we did TLC tests to total alkaloids. According to the results of tests, we isolated total alkaloids by silica gel column chromatography, respectively used benzoLacetidin:diethylamine=7:2:1 and acetidinxhloroformrdiethylamine-8:1:1 to elute and got two alkaloid monomers: CJ- I and CJ- II. Through chemical and spectral methods, we knew that CJ- I may be acanthoine. CJ- II was not identified because of its little quantity.In the third part, we studied the extraction, decontamination, isolation and formula weight assaying of polysaccharides from Cirsium japonicum DC.. The polysaccharides were extracted with hot water, precipitated by alcohol, scrubbed precipitant with 95%EtOH and acetone, then vacuum dried to get coarse polysaccharides (CJ). There are lots of impurities in water extracted polysaccharides, such as protein and inorganic salt. Their existences added difficulties of polysaccharide’s isolation. So we must get rid of them. Protein in the precipitants was removed by Sevag method. Inorganic salt was removed by dialysis method. The effects are good. CJ was still a mixture after deprotein and dialysis. It needed isolating further according to formula weight. The products were isolated with DEAE-cel
【Key words】 Cirsium japonicum DC.; alkaloids; polysaccharides; antioxygenic activities; bacteriostatic activities;
- 【网络出版投稿人】 陕西师范大学 【网络出版年期】2004年 01期
- 【分类号】TQ461
- 【被引频次】10
- 【下载频次】708