【作者】 陆英；

【导师】 刘仲华；

【作者基本信息】 湖南农业大学 ， 茶学， 2012， 博士

【摘要】 为进一步研究植物中化学成分及其生理活性,本论文对紫锥菊多种化学成分进行制备分离、活性筛选及紫甘薯中主要化学成分的制备分离,取得主要结果如下：1、采用高速逆流色谱与制备液相色谱技术制备分离得到紫锥菊中13个高纯度化合物,经鉴定,3个为新化合物：2-O-异阿魏酰基酒石酸、2-O-咖啡酰基-3-O-异阿魏酰酒石酸、及2,3-O-双异阿魏酰基酒石酸。2、采用高速逆流色谱技术从国产紫甘薯中分离制备得到4个化合物,经质谱、核磁共振、紫外等技术进行结构鉴定,分别为3-O-{6-O-(E)-咖啡酰-2-O-[6-O-(E)-对羟基苯甲酰-β-D-吡喃葡糖基]-β-D-吡喃葡糖苷}-[5-O-(β-D-吡喃葡糖苷)]芍药素、3-O-{6-O-(E)-咖啡酰-2-O-[6-O-(E)-阿魏酰-β-D-吡喃葡糖基]-β-D-吡喃葡糖苷}-[5-O-(β-D-吡喃葡糖苷)]芍药素、3-O-咖啡酰基奎宁酸,3,5-O-二咖啡酰基奎宁酸。3、采用流感病毒神经氨酸酶抑制模型研究了紫锥菊单体化合物及不同规格提取物对流感病毒A/PR/8/34(H1N1)神经氨酸酶的抑制作用。结果表明,7个肉桂酸类化合物均显示出显著抑制作用,其中菊苣酸的抑制作用最强(IC50=5.68±0.19μg/mL),表明紫锥菊具有潜在的抗病毒活性。4、采用磺酰罗丹明B染色法研究了紫锥菊单体化合物及不同规格提取物对人肝癌BEL-7402及人乳腺癌MCF-7细胞株的抑制作用。结果表明,肉桂酸类化合物及烷基酰胺单体化合物对肿瘤细胞抑制作用较弱,但烷基酰胺类混合物对两种肿瘤细胞的抑制作用较强,半抑制浓度IC50分别为0.05mg/mL、0.03mg/mL。5、采用趋化因子受体拮抗剂筛选模型研究了紫锥菊单体化合物及不同规格提取物对趋化因子受体CXCR4、CCR5的拮抗作用。结果表明,各样品仅具有较弱的拮抗作用,表明紫锥菊对HIV病毒侵入宿主细胞时的抑制作用不明显。6、研究了紫锥菊单体化合物及不同规格提取物对金黄色葡萄球菌、大肠杆菌、沙门氏菌、溶血性链球菌的抑制作用。结果表明各样品均无抑制作用。7、对紫锥菊肉桂酸类化合物进行体外抗氧化作用及其构效关系研究,结果表明化合物抗氧化性强弱基本表现为：菊苣酸、2-O-咖啡酰基酒石酸、2-O-咖啡酰基-3-O-阿魏酰酒石酸、2,3-O-双异阿魏酰基酒石酸、2-O-异阿魏酰基酒石酸；体外抗氧化活性大小与咖啡酸苯环上的酚羟基位置、数目相关,其中对位酚羟基的活性最强,邻位羟基较弱；对DNA损伤均具有显著保护作用,但构效关系不明显。8、建立了HPLC-PDA方法同时检测紫锥菊中7个肉桂酸及5个烷基酰胺类化合物的方法,通过梯度洗脱于60min内可在C18反相色谱柱上完全分离,样品预处理简单,可作为紫锥菊E.purpurea中活性成分的标准检测方法。更多还原

【Abstract】 This study has two parts which one deals mainly with chemical, bioactive investigation and HPLC detection method on Echinacea purpurea (L.) Moench and another deals with chemical on Purple sweet potato, the main results are as follows:1、Set up High-speed countercurrent chromatography and preparative HPLC methods to separate cinnamic acids and alkylamides in Echinacea purpurea, fifteen high-purity compounds were got from Echinacea purpurea and thirteen structures were identified by MS, NMR, UV, three compounds such as2-O-isoferuloyltartaric acid,2-O-caffeoyl-3-O-isoferuloyltartaric acid,2,3-di-O-isoferuloyltartaric acid were new compounds.2、Set up a High-speed countercurrent chromatography mothed to separate four compounds from purple sweet potato, the structure of them identified by MS, NMR, UV, the resuls showed two of them were acylated anthocyanins and anther were chlorogenic acid and ischlorogenic acid A.3、The model of inhibition influenza virus neuraminidase enzyme has been used to study the compounds in Echinacea purpurea on the neuraminidase of influenza virus A/PR/8/34(H1N1). The results showed that cinnamic acids have significant inhibitions, cinnamic acid compound3was the strongest and IC50values was5.68±0.19μg/mL. Suggesting that one pathway of antiviral activity of Echinacea purpurea was the inhibition of virus neuraminidase enzyme.4、Sulfonyl rhodamine B staining method has been used to study the compounds of Echinacea purpurea on BEL-7402human hepatoma and MCF-7human breast cancer cell lines. The results showed that cinnamic acids and alkylamide compounds have weak inhibition on tumor cell, while the mixture of alkylamides were strong inhibition compare with purity compound, IC50values were0.05mg/mL and0.03mg/mL respectively.5、The model of antagonism has been used to study the compounds of Echinacea purpurea on chemokine receptor CXCR4and CCR5. The results showed that all the samples only have a weak antagonism, suggesting it was not obviously that Echinacea against HIV in the infection of host cells.6^Study the inhibition of compounds of Echinacea purpure on four kinds of bacteria. The results showed there were no inhibition on bacteria such as Staphylococcus aureus, Escherichia coli, Salmonella and hemolytic streptococcus.7、Chemical method have been used to study antioxidant activities of cinnamic acid derivatives of Echinacea purpure, the results showed that the antioxidant of compounds were as follows:Cichoric acid,2-caffeoyltartaric acid,2-caffeoyl-3-feruloyltartaric acid,2,3-diisferuloyltartaric acid,2-isferuloyltartaric acid; Structure-activity relationship showed:active groups of antioxidantactive was the phenolic hydroxyl on benzene of caffeic acide groups, and the bit phenolic hydroxyl was the strongest activity and adjacent was weak, but the protective effect of DNA damage does not exist this relationship, indicating that the protective effect has different action mechanism.8、Simultaneous analysis7cinnamic acid compounds and5alkylamide compounds in Echinacea purpure by HPLC-PDA with gradient elution on a C18reverse phase column in60minutes without the pretreatment of samples, it was suitable as standard analytical method to detect the active ingredients in Echinacea purpure.更多还原