臺灣博碩士論文加值系統

在台灣田野裡，一種快速蔓生的雜草，扛板歸 (Polygonum perfoliatum) ，曾被發現其具有增加白血球及血小板的功用，但對於其有效成分卻不清楚。近年來一些學者的研究報告指出人體內過多的自由基與諸多疾病有直接或間接有關。因此本研究專以扛板歸為材料，探討甲醇萃取物中有效清除自由基的化合物及其生物活性。從研究結果顯示扛板歸全草萃取物經分配層析後共取得乙酸乙酯層、正丁醇層及水層，其中以乙酸乙酯層和正丁醇層對清除自由基的效果最好而乙酸乙酯層經純化後共得九種化合物，再經DPPH自由基測試後，共得兩種有效成分，分別為α-tocopherol和methyl trans-ferulate, 其在207 μg/ml的濃度時，清除自由基的能力分別為96 % 和94 %，在另一項植物相剋作用潛能研究的試驗中，利用紙色層分析法和高效能液相層析法從扛板歸水溶萃取液中之乙醚層得知至少有九種酚酸化合物，其中已鑑得有caffeic acid、p-coumaric acid、p-hydroxybenzoic acid、protocatechuic acid 及vanillic acid,此類酚酸化合物均是已知者，其植物相剋作用的潛能已被證實。然而，在溫室盆栽實驗中，扛板歸粉末與土壤混合後，其比例在1 % 至4 % 間，卻發現扛板歸粉末，具有促進鬼針草 ( Bidens pilosa ) 生長的效果，此顯示扛板歸亦具植物相生的功能。
上述結果顯示扛板歸具有抗氧化能力及具植物相生相剋之潛能，此天然物有待進一步深入探討。

n the field of Taiwan, there is a vine-type weed, Polygonum perfoliatum , which has been found to have a function of raising white blood cells and platelet counts. But its effective composition is still unknown. Recently the researcher reported that the excess free radicals can directly or indirectly cause many diseases of human beings. For this purpose, Polygonum perfoliatum was chosen as material to study the components efficacy of scavenger DPPH free radical and its biological activity of natural products isolated from the methanol extract. By means of chromatography, the ethylacetate layer, n-butanol layer and water layer were obtained. Among which the ethylacetate layer, and n-butanol layer revealed a better result in the efficacy of scavenging radicals, while the ethylacetate layer was further purified into 9 components, each of them also passed through the test of scavenging DPPH free radicals. At the concentration of 207 μg/ml, only two purified components of α-tocopherol and methyl trans-ferulate were confirmed with the efficacy of scavenging DPPH free radicals with 96 % and 94 %, respectively. On the other hand, the allelopathic experiments was also carried out by using the compounds isolated from water extracts of the weed. The allelopathic components were isolated from the ether layer of water extracts and 9 phenolic acids were identified by paper and high performance liquid chromatographies. These compounds include caffeic acid, p-coumaric acid, p-hydroxybenzoic acid, protocatechuic acid, and vanillic acid, which have been confirmed to be allelopathic. In addition, in a greenhouse pot experiments, the powder of the dry weed was mixed with soil by a ratio of 1 %、2 %、3 %、and 4 % to that the test of Bidens pilosa, the results revealed a stimulative effect.
From the above results it is concluded that the weed, Polygonum perfoliatum, has the potential of biological activities, including antioxidation and allelopathy. The responsible natural products should be further studied hereafter.

中文摘要......................................................................................................... I
英文摘要..........................................................................................................II
誌謝............................ ....................................................................................IV
目錄..................................................................................................................V
圖表目錄………………………………………………………………..…VIII
壹、前言.............................................................................................................1
貳、文獻整理....................................................................................................4
一、何謂自由基.........................................................................................4
(一) 自由基的由來及影響..................................................................4
(二) 自由基對生物影響......................................................................4
(三) 自由基的清除與抗氧化劑作用..................................................5
1. 酵素系統..................................................................................5
2. 非酵素系統..............................................................................7
二、扛板歸研究簡介.............................................................................10
三、植物相剋作用潛能………………………………….…...….…....13
(一) 植物相剋作用............................................................................13
1. 自毒作用................................................................................13
2. 毒他作用................................................................................13
　　 (二) 植物相剋物質..........................................................................14
　　 (三) 相剋物質的釋出......................................................................14
　　 (四) 相剋物質對作物的影響..........................................................15
　　 (五) 相剋作用在農業上的應用......................................................15
叁、材料與方法.............................................................................................17
一、扛板歸中化合物分離與純化.......................................................17
(一) 實驗藥品與儀器......................................................................17
1. 實驗材料來源........................................................................17
2. 化學藥品................................................................................17
3. 儀器........................................................................................17

(二) 實驗方法....................................................................................18
1. 材料製備................................................................................18
2. 甲醇萃取物的純化與分析....................................................18
3. 正丁醇萃取物分離................................................................19
二、扛板歸萃取物之清除自由基實驗.................................................20
(一) 實驗藥品與儀器........................................................................20
1. 材料來源................................................................................20
2. 化學藥品................................................................................20
3. 儀器........................................................................................20
(二) 實驗方法....................................................................................20
1. 清除DPPH自由基活性實驗...............................................20
2. 抗氧化活性的測定...............................................................20
三、扛板歸全草生物分析....................................................................22
(一) 實驗藥品與儀器.......................................................................22
1. 材料來源...............................................................................22
2. 化學藥品...............................................................................22
3. 儀器.......................................................................................22
(二) 實驗方法...................................................................................22
1. 水溶萃取液的製備...............................................................22
2. 萃取液的種子生物分析.......................................................23
3. 酚類化合物的萃取法...........................................................23
4. 濾紙色層分析法...................................................................23
5. 濾紙色層之種子生物分析...................................................24
6. 溫室盆栽實驗.......................................................................24
肆、結果與討論............................................................................................25
一、扛板歸全草甲醇萃取物乙酸乙酯層之成分分離…....................25
(一) 扛板歸全草甲醇萃取物乙酸乙酯層分離結果.......................25
(二) 化合物摘要...............................................................................26
(三) 化合物之分析與鑑定...............................................................30
二、扛板歸抗氧化能力之測定結果....................................................65
(一) 清除DPPH自由基能力測定...................................................65
1. 不同溶劑萃取法其各分配層析區分物清除DPPH
自由基效果比較...................................................................65
2. 扛板歸甲醇萃取各極性區分物清除DPPH自由基效果
比較.........................................................................................67
3. 自扛板歸乙酸乙酯層中分離純化出之化合物清除
DPPH自由基測試................................................................69
(二) 抑制脂質過氧化活性測定結果................................................73
1. 扛板歸不同溶劑萃取法其各分配層析區分物抑制脂質
過氧化之效果........................................................................ 73
2. 扛板歸甲醇萃取純化之化合物抑制脂質過氧化之
效果.........................................................................................75
三、水溶萃取液pH值、滲透壓、濾紙層析以及種子生物
分析.................................................................................................77
(一) 水溶萃取液pH值之比較.........................................................77
(二) 水溶萃取液滲透壓值之比較....................................................78
(三) 水溶萃取液之生物分析結果....................................................79
(四) 濾紙色層結果之種子生物分析................................................82
(五) 酚類化合物的萃取與偵測........................................................84
(六) 溫室盆栽實驗............................................................................89
伍、結論.........................................................................................................93
參考文獻.........................................................................................................94
作者簡介.......................................................................................................103

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