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研究生:陳勇智
研究生(外文):Chen Yong Zhi
論文名稱:虎杖萃取液的抗氧化與抑制酪氨酸酵素之研究
論文名稱(外文):Study on Anti-oxidative and Anti-tyrosinase Activities of Polygonum Cuspidatum Extracts
指導教授:張耀南張耀南引用關係
指導教授(外文):Chang Yaw Nan
學位類別:碩士
校院名稱:大葉大學
系所名稱:生物產業科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:70
中文關鍵詞:虎杖白藜蘆醇抗氧化酪氨酸酵素
外文關鍵詞:Polygonum cuspidatumresveratrolanti-oxidative activityanti-tyrosinase
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本研究以 100% 純水、10% 與70% 乙醇分別萃取傳統中草藥虎杖中有效成分-白藜蘆醇,並進行萃取液中白藜蘆醇的定量分析,及其抗氧化與抑制酪氨酸酵素之評估。實驗結果顯示,以 70% 乙醇萃取所得冷凍乾燥萃取物含量 ( 30% 萃取率 ) 與白藜蘆醇含量 ( 1.57% 乾重 ) 為最高。在 DPPH 抗氧化試驗中,除了最佳抗氧化之冷凍乾燥 70% 乙醇萃取液之外,其它萃取液之抗氧化活性隨其添加濃度或白藜蘆醇含量之增加而先行增加,而後隨其濃度或含量增加而漸降低其活性,冷凍乾燥 70% 乙醇萃取液隨其添加濃度或白藜蘆醇含量之增加而先行略降低其抗氧化活性,而後有漸增加其活性之趨勢。在抑制酪氨酸酵素活性方面,除了熱烘乾燥 100% 純水與 10% 乙醇萃取液可維持 80%~90% 酪氨酸酵素相對活性外,仍以冷凍乾燥 70% 乙醇萃取液的抑制酪氨酸酵素效果為最佳,但其添加濃度必須超過1000g/mL或白藜蘆醇含量超過 15.7 g 時,其酪氨酸酵素相對活性將由原有 70% 逐漸下降,換言之,其抑制酪氨酸酵素效果則逐漸明顯而有效。由此可知,以 70% 乙醇萃取虎杖可得到最高冷凍乾燥萃取物含量與白藜蘆醇含量,其抗氧化活性與抑制酪氨酸酶功效為最佳。
The resveratrol contents, anti-oxidative ( DPPH-free-radical-scavenging effect ) and anti-tyrosinase activities of Polygonum cuspidatum ( one of Chinese traditional medicine ) extracts with 100% pure water ( Wes ) and 10% and 70% ethanol solutions ( 0.1 EtEs and 0.7 EtEs, respectively ) were investigated in this study. The maximum extract content ( 30% extraction yield ) and resveratrol ( 1.57% dried weight ) was obtained from the freeze-dried 0.7 EtEs of P. cuspidatum. Among the P. cuspidatum extracts, only the freeze-dried 0.7 EtEs had strong DPPH-scavenging effects or anti-oxidative activities, while the activities of the freeze-dried 0.7 EtEs first gradually decreased to a certain value as increasing their addition concentrations or resveratrol contents, and then increased as increasing the concentrations or contents. For the other P. cuspidatum extracts, the anti-oxidative activities first increased and then decreased to some certain values as increasing their addition concentrations or resveratrol contents. Except that the tyrosinase relative activities of the hot-dried WEs and 0.1 EtEs were kept at 80%~90%, the freeze-dried 0.7 EtEs also had stronger anti-tyrosinase activities than the others. However, the tyrosinase relative activities started decreasing from original 70% or the anti-tyrosinase activities became much significant and effective when the addition concentrations or resveratrol contents of the freeze-dried 0.7 EtEs were over 1000 g/mL or 15.7 g, respectively. In this study, only the freeze-dried 0.7 EtEs had the higher extract and resveratrol contents and stronger anti-oxidative anti-tyrosinase activities than the others.
目錄

封面內頁
簽名頁
授權書 iii
中文摘要 iv
英文摘要 v
誌謝 vi
目錄 vii
圖目錄 ix
表目錄 xi

1. 前言 1
2. 文獻回顧 4
2.1 虎杖.. 4
2.2 白藜蘆醇 7
2.2.1 抗氧化活性 18
2.2.2 抗發炎及抗血小板凝集作用 19
2.2.3 抗癌作用 20
2.2.4調節血管舒張活性 21
2.2.5 具有雌性激素活性 22
2.2.6 抗衰老、抗疲勞作用 22
2.2.7 臨床應用 22
2.2.8 抑菌性 23
2.3自由基 23
3. 材料與方法 26
3.1實驗材料及方法 26
3.1.1實驗材料 26
3.1.2儀器設備 26
3.2虎杖萃取液的製備 27
3.3中草藥虎杖萃取液抗氧化能力測定 28
3.4中草藥虎杖抑制酪氨酸酵素的測定 29
3.5反式白藜蘆醇 HPLC 定量分析條件 32
4. 結果與討論 33
4.1虎杖萃取率 33
4.2虎杖抗氧化能力試驗 33
4.3抑制酪氨酸酵素能力測定 35
4.4 HPLC定量白藜蘆醇 37
5. 結論 50
參考文獻 52

圖目錄

圖 2.1 藥用植物圖鑑-虎杖 6
圖 2.2 反式白藜蘆醇結構式 9
圖 4.1 不同虎杖萃取液的 DPPH 清除率 34
圖 4.2 冷凍乾燥虎杖萃取物之酪氨酸酶相對活性 ( % ) 36
圖 4.3 熱烘乾燥虎杖萃取物之酪氨酸酶相對活性 ( % ) 36
圖 4.4 反式白藜蘆醇的分光光度 ( 200-500 nm ) 吸收光譜 38
圖 4.5 2.20 μg/mL 反式白藜蘆醇之 HPLC 分析圖 39
圖 4.6 1.10 μg/mL 反式白藜蘆醇之 HPLC 分析圖 39
圖 4.7 0.55 μg/mL 反式白藜蘆醇之 HPLC 分析圖 39
圖 4.8 0.2750 μg/mL 反式白藜蘆醇之 HPLC 分析圖 40
圖 4.9 0.1375 μg/mL 反式白藜蘆醇之 HPLC 分析圖 40
圖 4.10 反式白藜蘆醇的 HPLC 標準曲線 41
圖 4.11 150 μg/mL 冷凍乾燥虎杖水萃取物之 HPLC 分析圖 42
圖 4.12 150 μg/mL 冷凍乾燥虎杖 10% 乙醇萃取物之 HPLC 分析圖 42
圖 4.13 150 μg/mL 冷凍乾燥虎杖 70% 乙醇萃取物之 HPLC 分析圖 43
圖 4.14 150 μg/mL 熱烘乾燥虎杖水萃取物之 HPLC 分析圖 43
圖 4.15 150 μg/mL 熱烘乾燥虎杖 10% 乙醇萃取物之 HPLC 分析圖 44
圖 4.16 150 μg/mL 熱烘乾燥虎杖 70% 乙醇萃取物之 HPLC 分析圖 44
圖 4.17 750 μg/mL 冷凍乾燥虎杖水萃取物之 HPLC 分析圖 45
圖 4.18 750 μg/mL 冷凍乾燥虎杖水萃取物含反式白藜蘆醇 0.55 μg/mL 之 HPLC 分析圖 45
圖 4.19 750 μg/mL 熱烘乾燥虎杖水萃取物之 HPLC 分析圖 46
圖 4.20 750 μg/mL 熱烘乾燥虎杖水萃取物含反式白藜蘆醇 0.55 μg/mL 之 HPLC 分析圖 46
圖 4.21 虎杖萃取物中反式白藜蘆醇含量與 DPPH 清除率之關係圖 49
圖 4.22 虎杖萃取物中反式白藜蘆醇含量與酪氨酸酵素相對活性之關係圖 49

表目錄

表 2.1 不同紅酒商品反式白藜蘆醇含量 12
表 2.2 不同白酒商品反式白藜蘆醇含量 14
表 2.3 不同農業資源中反式白藜蘆醇含量 16
表 2.4 反式白藜蘆醇之主要生物活性性質 17
表 3.1 酪氨酸酶的試驗組合設計 31
表 4.1 不同虎杖萃取物濃度 ( μg/mL ) 中反式白藜蘆醇含量 ( μg ) 對照表 48
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