臺灣博碩士論文加值系統

本篇研究目標為由天然物分離得到具酪胺酸酶抑制及抗氧化活性之化合物，預試驗發現天然物鴨舌癀（Phyla nodiflora）地上部之甲醇萃取物有總酚類含量及良好的抗酪胺酸酶及自由基清除之活性，進行成分分離後得到五個已知化合物，分別為eupafolin（1）、hispidulin（2）、larycitrin（3）、β-sitosterol（4）及stigmasterol（5）；將eupafolin（1）及hispidulin（2）進行抑制酪胺酸酶實驗，發現皆具有良好的抑制效果，其中針對較強力活性的eupafolin（1）（IC50 = 56.0 μM）進行酵素動力學實驗結果發現，eupafolin（1）為競爭型抑制酪胺酸酶；自由基清除能力試驗方面，將eupafolin（1）及hispidulin（2）進行1,1-diphenyl-2-picryhydrazyl（DPPH）、2,2''-azinobis-3-ethylbenzothiazoline-6-sulfonic acid（ABTS）及超氧陰離子（superoxide anion）自由基清除活性試驗，hispidulin（2）對於DPPH•及 ABTS•＋自由基有清除效果，而eupafolin（1）對三種自由基都有優良的清除活性，在黃嘌呤氧化酶酵素動力學實驗結果顯示，eupafolin（1）是以混合型抑制模式抑制黃嘌呤氧化酶。

The goal of our research is to conduct bioactive compounds with tyrosinase inhibitory and antioxidative activities from the aerial part of Phyla nodiflora. The methanolic extracts of Phyla nodiflora showed potential total phenolic contents, tyrosinase inhibitory and free radical scavenging effect. In our continuous research, five compounds including eupafolin (1), hispidulin (2), larycitrin (3), β-sitosterol (4) and stigmasterol (5) were isolated. In antityrosinase assay, both eupafolin (1) and hispidulin (2) had good activity. Eupafolin (1) showed stronger activity (IC50 = 56.0 μM). In kinetic experiment, we found that eupafolin (1) showed competitive inhibition mode with tyrosinase. In radical scavenging assays, hispidulin (2) had good effect on both 1,1-diphenyl-2-picryhydrazyl (DPPH) and 2,2''-azinobis-3-ethylbenzo-
thiazoline-6-sulfonic acid (ABTS) radicals while eupafolin (1) had very good scavenging effect on DPPH, ABTS and superoxide anion radicals. Furthermore, we realized that eupafolin (1) inhibits xanthine oxidase with a mixed-type inhibition mode.

目錄............................................................................................................i
圖目錄.......................................................................................................iv
表目錄...................................................................................................... vi
中文摘要....................................................................................................1
Abstract.......................................................................................................2
第一章 緒論................................................................................................3
第一節 研究背景.................................................................................3
第一項 馬鞭草科植物介紹..........................................................3
第二項 鴨舌癀植物介紹..............................................................5
第二節 動機與目的...........................................................................19
第二章 研究方法......................................................................................23
第一節 儀器與材料...........................................................................23
第二節 生物活性試驗篩選...............................................................29
第一項 總酚類含量之評估........................................................29
第二項 抑制酪胺酸酶活性試驗................................................31
第三項 酪胺酸酶酵素動力學試驗（Linweaver-Burk plots法）.33
第四項 自由基清除能力之評估................................................35


第五項 黃嘌呤氧化酶酵素動力學試驗
（Linweaver-Burk plots法）.........................................39
第三章 結果與討論..................................................................................41
第一節 鴨舌癀地上部各極性劃分層活性試驗結果與討論...........41
第一項 各極性劃分層之分離...................................................41
第二項 鴨舌癀萃取物總酚類含量試驗...................................43
第三項 鴨舌癀萃取物酪胺酸酶抑制活性試驗.......................44
第四項 鴨舌癀萃取物清除ABTS•＋自由基活性實驗結果...46
第五項 鴨舌癀萃取物清除DPPH•自由基活性實驗結果.....49
第六項 鴨舌癀萃取物清除超氧自由基活性實驗結果...........52
第二節 化合物活性試驗結果與討論…...........................................55
第一項 化合物之純化分離......................................................55
第二項 Eupafolin（1）之結構證明...........................................57
第三項 Hispidulin（2）之結構證明..........................................61
第四項 Larycitrin（3）之結構證明...........................................65
第五項 β-Sitosterol（4）及Stigmasterol（5）之結構證明...67
第六項 化合物酪胺酸酶抑制活性試驗結果與討論..............69
第七項 化合物清除ABTS•＋自由基活性實驗結果...............75
第八項 化合物清除DPPH•自由基活性實驗結果.................76
第九項 化合物清除超氧自由基活性實驗結果......................77
第四章 結論…………............................................................................82
第五章 化合物物理化學性質................................................................85
第六章 參考文獻......................................................................................89

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