金边瑞香活性成分提取分离及抗氧化活性研究
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摘要
金边瑞香(Daphne odora Var.marginata)系瑞香科瑞香属常绿小灌木,具有观赏价值以及多种利用价值,可作造纸原料,可提取芳香油。民间常用金边瑞香叶捣烂敷患处,治疗溃烂、无名肿毒、乳腺炎等;其花在白酒中浸泡后可用于治疗牙痛、风湿性关节炎以及因损伤引起的肿痛和各种无名疼痛等。近代医学研究表明,金边瑞香具有抑菌、抗炎、抗风湿、耐缺氧、抗脂质过氧化、延缓衰老和调节免疫等药理活性。本论文对金边瑞香的化学成分进行了初步研究,并采用各种化学及生物模型考察了其抗氧化活性,以期为金边瑞香的开发利用提供理论基础。本论文共分七个章节,主要内容如下:
第一章绪论部分,综述了近年来国内外研究体外抗氧化及清除自由基能力几种化学模型的原理及应用;概述了金边瑞香化学成分及药理活性研究进展。
第二章采用溶剂萃取法对全植株金边瑞香原料进行组分化提取。
第三章采用五种化学模型评价了金边瑞香四种提物清除超氧自由基、DPPH自由基、羟基自由基、还原铁离子以及鳌合亚铁离子的能力。测定了各组分的总黄酮含量,并探讨了总黄酮含量与各活性之间的相关性。
第四章通过薄层色谱对洗脱溶剂进行筛选,采用硅胶柱层析对活性较强的乙酸乙酯提取物进行分离,得到三个子组分,采用反相高效液相色谱法分别测定了金边瑞香乙酸乙酯组分以及三个子组分的化学组成及其含量。
第五章采用六种化学和生物模型评价了金边瑞香乙酸乙酯提取物及其子组分清除超氧自由基、DPPH自由基、羟基自由基、还原铁离子、鳌合亚铁离子的能力以及对羟基自由基引起的DNA损伤的保护作用。初步探讨了化学组成及含量与各活性之间的相关性。同时测定了各组分的总黄酮、总酚含量,并讨论了它们与各活性之间的相关性。
第六章采用Sephadex LH-20柱层析对活性较强的组分F2进行分离纯化,通过制备HPLC得到两个单体化合物。
第七章对整个实验结果进行了总结,对今后的实验和研究进行了展望。
Daphne odora var.marginata,family of Thymelaeaceae,is a famous ornamental flower and medical plant,also used as paper making and aromatic oil extracting.The pounded leaves could applied at the afflicated part,utilized as ulcerate,gall and mastitis treatment.After marinated in wine,the flowers obtain the ability to cure toothache,rheumatoid arthritis,gall caused by gall and verious ache.Recent pharmacological studies indicated that Daphne odora var.marginata had anti-lipid peroxidation,analgesic,immunoloregulation,anti-inflammation,antibacterial, anti-hypoxia and anti-tumor bioactivity.In this thesis,separation and purification of the chemical constituents and assaying about the antioxidant activity of Daphne odora var.marginata were studied.For the sake of founding the theoretical foundation for the depth development and utilization of Daphne odora var.marginata.The thesis were divided into six chapters,the main contents of each chapter was as follows:
Chapter 1.Introduction of the mechanism and application of five antioxidant and free radical scavenging assays;summarize about the chemical constituents and medical activiteies of daphne odora var.marginata.
Chapter 2.Extracting the crude plant of daphne odora var marginata by normal solvent with partition.
Chapter 3.Assayed the activities of daphne odora var.marginata by using superoxide radical scavenging,DPPH radical scavenging,hydroxide radical scavenging,ferrous ion chelating and ferric ion reducing.Measured the total flavonoid contents of each fraction,compared the activities with the total flavonoid contents.
Chapter 4.Seperation and purification of the acetic ether extract with high activity by silica gel column chromatography after choosing eluent by TLC,three subfractions were collected. The chemical constituents of EA and its subfractions were performed on RP-HPLC.
Chapter 5.Six chemical and biological assays were performed on evaluting the activities of EA and its three subfractions.They were superoxide radical scavenging,DPPH radical scavenging,hydroxide radical scavenging,ferrous ion chelating,ferric ion reducing,protective effect on damaged DNA caused by hydroxide radical.Investigated the coefficient between chemical constituents and contents and verious activities.The total flavonoid and phenolic contents were also measured,and their coefficient with different activities were estimated too.
Chapter 6.Isolation and purification the active fraction F2 by sephadex LH-20 column chromatography,two active compounds obtained.It need a furder structural identification and bioactive detection.
Chapter 7.Summarized the entire experimental results.Prospected the experiment and research in the future.
第一章绪论部分,综述了近年来国内外研究体外抗氧化及清除自由基能力几种化学模型的原理及应用;概述了金边瑞香化学成分及药理活性研究进展。
第二章采用溶剂萃取法对全植株金边瑞香原料进行组分化提取。
第三章采用五种化学模型评价了金边瑞香四种提物清除超氧自由基、DPPH自由基、羟基自由基、还原铁离子以及鳌合亚铁离子的能力。测定了各组分的总黄酮含量,并探讨了总黄酮含量与各活性之间的相关性。
第四章通过薄层色谱对洗脱溶剂进行筛选,采用硅胶柱层析对活性较强的乙酸乙酯提取物进行分离,得到三个子组分,采用反相高效液相色谱法分别测定了金边瑞香乙酸乙酯组分以及三个子组分的化学组成及其含量。
第五章采用六种化学和生物模型评价了金边瑞香乙酸乙酯提取物及其子组分清除超氧自由基、DPPH自由基、羟基自由基、还原铁离子、鳌合亚铁离子的能力以及对羟基自由基引起的DNA损伤的保护作用。初步探讨了化学组成及含量与各活性之间的相关性。同时测定了各组分的总黄酮、总酚含量,并讨论了它们与各活性之间的相关性。
第六章采用Sephadex LH-20柱层析对活性较强的组分F2进行分离纯化,通过制备HPLC得到两个单体化合物。
第七章对整个实验结果进行了总结,对今后的实验和研究进行了展望。
Daphne odora var.marginata,family of Thymelaeaceae,is a famous ornamental flower and medical plant,also used as paper making and aromatic oil extracting.The pounded leaves could applied at the afflicated part,utilized as ulcerate,gall and mastitis treatment.After marinated in wine,the flowers obtain the ability to cure toothache,rheumatoid arthritis,gall caused by gall and verious ache.Recent pharmacological studies indicated that Daphne odora var.marginata had anti-lipid peroxidation,analgesic,immunoloregulation,anti-inflammation,antibacterial, anti-hypoxia and anti-tumor bioactivity.In this thesis,separation and purification of the chemical constituents and assaying about the antioxidant activity of Daphne odora var.marginata were studied.For the sake of founding the theoretical foundation for the depth development and utilization of Daphne odora var.marginata.The thesis were divided into six chapters,the main contents of each chapter was as follows:
Chapter 1.Introduction of the mechanism and application of five antioxidant and free radical scavenging assays;summarize about the chemical constituents and medical activiteies of daphne odora var.marginata.
Chapter 2.Extracting the crude plant of daphne odora var marginata by normal solvent with partition.
Chapter 3.Assayed the activities of daphne odora var.marginata by using superoxide radical scavenging,DPPH radical scavenging,hydroxide radical scavenging,ferrous ion chelating and ferric ion reducing.Measured the total flavonoid contents of each fraction,compared the activities with the total flavonoid contents.
Chapter 4.Seperation and purification of the acetic ether extract with high activity by silica gel column chromatography after choosing eluent by TLC,three subfractions were collected. The chemical constituents of EA and its subfractions were performed on RP-HPLC.
Chapter 5.Six chemical and biological assays were performed on evaluting the activities of EA and its three subfractions.They were superoxide radical scavenging,DPPH radical scavenging,hydroxide radical scavenging,ferrous ion chelating,ferric ion reducing,protective effect on damaged DNA caused by hydroxide radical.Investigated the coefficient between chemical constituents and contents and verious activities.The total flavonoid and phenolic contents were also measured,and their coefficient with different activities were estimated too.
Chapter 6.Isolation and purification the active fraction F2 by sephadex LH-20 column chromatography,two active compounds obtained.It need a furder structural identification and bioactive detection.
Chapter 7.Summarized the entire experimental results.Prospected the experiment and research in the future.
引文
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