野菊花有效组分的鉴定特征研究

【作者】 汪国鹏；

【导师】 张贵君；

【作者基本信息】 北京中医药大学 ， 中药学， 2006， 硕士

【摘要】 1.目的野菊花是菊科植物野菊(Chrysanthemum indicum L.)的干燥头状花序,其主要药理活性成分为总黄酮,具有清热解毒、降血压、抗病毒、抑制血小板凝聚等功能。野菊花是金菊粉针剂的主要药效成分,但是目前还不能说明其抗病毒的有效组分。本课题是旨在以总黄酮含量为指标找到野菊花的有效部位,然后对其有效部位的化学成分进行研究,从而为找到其药效组分奠定基础。2.方法及研究思路(1)研究方法1)采用正交试验的方法对野菊花总黄酮提取、富集工艺进行研究。2)采用现代色谱分离手段对有效组分的化合物进行分离、纯化。3)采用波谱和质谱法对化合物的结构进行鉴定。(2)技术路线野菊花水提取物富集总黄酮确定有效组分有效组分的化学单体分离、纯化化合物的结构鉴定3.内容第一部分:中药野菊花的化学成分研究、药理学研究进展综述第二部分:野菊花总黄酮富集工艺研究第三部分:野菊花有效部位化学组分的提取、分离和结构鉴定第四部分:总结与讨论4.结果(1)以野菊花总黄酮含量为指标,筛选其水提物的有效部位,最终确定的工艺流程为:野菊花用水提取3次,溶剂用量依次为10、10、10倍,提取温度100°C,提取时间为50,40,30min,水提液用80%乙醇醇沉后药液上大孔树脂柱,弃去水洗液和10%乙醇洗脱液,收集30%乙醇洗脱液,其中30%乙醇洗脱液就是最后我们得到的野菊花水提取物的有效部位。(2)利用现代色谱分离手段和波谱技术,对其有效部位进行化学成分研究,从中分离得到了13个单体化合物,利用波谱法和质谱法鉴定了其中11个化合物的结构,其中黄酮苷4个,即异槲皮苷(化合物3)、木犀草苷(化合物4)、秋英苷(化合物5)、蒙花苷(化合物6);游离黄酮3个,分别为圣草酚(化合物7)、木犀草素(化合物8)、槲皮素(化合物9);倍半萜化合物2个,为野菊花三醇A(化合物1)和野菊花四醇(化合物2);甾体1个,即胡萝卜苷(化合物11),还有一个为浙贝素(化合物10)。其中野菊花三醇A和野菊花四醇经文献检索为新化合物。3个为首次从该植物中分离得到,它们分别为异槲皮苷、圣草酚、浙贝素。5.结论及创新点(1)首次提出AB-8大孔树脂对野菊花的总黄酮进行富集的工艺。该工艺的优点是:大孔吸附树脂对总黄酮吸附效果较好,大孔树脂法富集野菊花总黄酮具有收率高,成本低,有机溶媒用量小,操作简便,工艺稳定,方法可行,可供大生产选用。(2)确定了野菊花水提取物药效组分中的11个化合物,初步确定其有效部位的药效组分的化合物特征为:黄酮类-萜类-木脂素衍生物类-甾体类,提出了其化学药效组分的质量评价的基本特征。(3)首次从野菊花中分离得到5个化合物,其中2个为新化合物,分别为:野菊花三醇A和野菊花四醇。6.讨论及展望野菊花水提取物中的药效组分大多数为黄酮类化合物,证明了其主要药效组分为黄酮类。有人从野菊花中分离得到过倍半萜类成分-野菊花醇,并且已经证明其有非常好的抗炎、抗病毒的药理活性,所以本次研究得到的新的倍半萜类成分野菊花三醇A和野菊花四醇是否具有抗病毒的活性,非常值得我们去进一步来研究。中药质量的评价应以组分去评价,这样可以客观地表述其本质特征。更多还原

【Abstract】 1.ObjectivesFlos Chrysanthemi Indici is dried capitulum of Chrysanthemum indicum L.. Total flavonoids as pharmacological active constituents have many activitives such as removing toxic heat、lowering the blood pressure、antivirus and Restraining platelet aggregation. Flos Chrysanthemi Indici is leading medicable ingredient of Jing Ju compound preparation,But we don’t know active chemical components of Flos Chrysanthemi Indici on antivirus by far. On the basis of the research on the chemical constituents of the effective fraction ,The purpose of our study is to establish the base for finding out a contact with chemical constituents and pharmacological activity further.2.Methods(1)Research method1) Study on the technics of extraction and concentration of total flavoninds with orthogonal test method.2) Study on the isolation and purification of chemical components by a few column chromatogram.3) Study on the structures of compounds on the basis of modern spectrum and MS method.(2) Technique lineWater extraction Concertrating the total flavoninds Confirming the effective fraction Isolating the chemical compounds from the effective fraction Identifying the structures of compounds.3.ContentsOur studies included four parts as follows: (1)The documents about the present research situation in Chinese medicine Flos Chrysanthemi Indici were summed up and analyzed.A review on the research progress of Flos Chrysanthemi Indici in chemical constituents and paharmacological activities was presented.(2) Study on the technics of extraction and concertration of total flavoninds.(3) The systemic study on the chemical constituents of the effective fraction.(4)Summarise the researching result、the academic meaning and valule of the researching results.4.Results(1) Taking the content of total flavonoids as observation index, we searched the effective fraction of water extract from Flos Chrysanthemi Indici .we have studied the extracti on processs by the L9(34) orthogonal design. The best technology is water as the extra cting solvent ,repeat 3 times, every time add 10、10、10 times water respectively and the extracting lasts 50,40,30min each time respectively.The liquid water-extration and alcohol-precipitation(80%ethanol) of Flos Chrysanthemi Indici was treated with a column of macroporous resin and eluted with distilled water、10%ethanol and 30%ethanol.The total flavonoids were enriched and purified in the eluent of 30%ethanol.(2) The chemical constituents of the effective fraction were investigated intensively. 13 compounds were isolated and purified by silica gel,polyamide,sephadex LH-20 column chromatogram. On basis of their spectral data,11of them were elucidate ed as chrysantetriolA,chrysanthetetrol,isoquercitrin,luteolin-7-O-β-D-glucoside, cosmosiin, linarin, eriodictyol, luteolin, quercetin, zhebeiresinol, daucosterol. Among them, chrysantetriolA and chrysanthetetrol are new compounds , isoquercitrin、eriodictyol and zhebeiresinol were obtained from Flos Chrysanthemi Indici for the fist time.5.Conlusions and Creative points(1) The enrichment and purification procedures of total flavonoids from Flos Chrysanthemi Indici with AB-8 macroporous resin absorption were established for the first timeThe concertrating processs have many virtues: with good adsorbability and small deadsorbability as well as low cost, little quantity organic solvent, the process is scientific and simple,the method is stabilization and fesible.(2) We have identified 11compounds from water extraction of flos chrysanthemi indici, and established their constituents characteristic preliminarily in sorts :flavonoids- terpenoids-lignans-steroides.On base of it we can supply the evidences for the quality moniter and control of flos chrysanthemi indici with basic feature.(3) Four compounds were obtained form flos chrysanthemi indici for the fist time ,Among them, chrysantetriolA and chrysanthetetrol are new compounds , isoquercitrin and zhebeiresinol were obtained for the fist time.6.Discussion and Expectation:The most of constituents that we have obtained from water extraction of flos chrysanthemi indici are flavonoids,so it’s believed that total flavonoids are the active components in the flos chrysanthemi indici are flavonoids.Chrysanthemol which had been obtained from flos chrysanthemi indici previously showed strong anti-inflammatory activity in mice,so it’s worth searching anti-inflammatory activity of chrysantetriolA and chrysanthetetrol as new eudesmane-type sesquiterpenes further.To clarify one’s esessential characteristic we should evaluate the quality of Chinese traditional medicine with chemical constituents.更多还原