臺灣博碩士論文加值系統

兒茶素(catechins)是茶葉中最主要的多酚類，其含量不僅影響茶湯的口味與品質，在先前的研究報告中更證實茶多酚在生化上具有抗癌、抗氧化、抗發炎、抗菌等功能。因此本篇論文以逆相高效能液體色層分析法(reverse-phase HPLC)分析亞洲各區域的不同茶種其茶多酚含量。並以水及不同濃度的酒精來萃取茶葉，發現75 %的酒精可萃取出最大量的茶多酚含量，大約是水萃取的1.53~10.52倍。在本實驗中，二十九種來自中國大陸、日本及臺灣的綠茶、烏龍茶、紅茶及普洱茶以沸水或75 %的酒精萃取並分析，發現其兒茶素的含量依序為綠茶(老葉)>綠茶(嫩葉)及烏龍茶>紅茶及普洱茶。我們不只分析茶葉成品，並於茶葉改良茶中親自摘取新鮮茶葉分析，發現老葉(五~七葉)的兒茶素含量大多高於嫩葉(一心二葉)，但相反的，嫩葉的咖啡因含量卻明顯的高於老葉。然而在分析綠茶成品當中，似乎也有類似的趨勢，嫩葉綠茶較一般綠茶含有較高的咖啡因及較少的兒茶素。同一種的茶樹，其茶花的咖啡因含量較少而兒茶素的含量卻和老葉差不多。同一種茶樹以不同的發酵程度製成品來比較，其咖啡因的含量依序為紅茶>烏龍茶>綠茶>鮮葉；而兒茶素的含量則依序為綠茶>烏龍茶>鮮葉>紅茶。
另外，我們以六種茶的成品及同一茶種的三個部份（老葉、嫩葉及花）的萃取物來比較其生物活性。在抗氧化的實驗中，普洱茶保護DNA受到氧自由基攻擊的能力最好，而所有的茶及茶花萃取物皆有不錯的抗氧化能力，其依序為普洱茶>綠茶>紅茶>烏龍茶。而抗發炎反應的實驗中，仍是以普洱茶的能力最好，且所有的茶及茶花萃取物也都有不錯的抗發炎效果。再來比較其引發細胞凋亡的能力，發現綠茶的效果最好，其效果強弱依序為綠茶>烏龍茶>紅茶>普洱茶。其鮮葉中的老葉也有不錯的引發細胞淍亡能力（相當於烏龍茶）。值得一提的是，以75 %酒精來萃取茶葉及茶花則比以沸水萃取有較好的生物活性。

An isocratic HPLC procedure was developed for simultaneous determination of caffeine and six catechins in tea and tea flower samples. A baseline separation was achieved on a Comsmosil C18-MS packed column with a solvent mixture of methanol/doubly distilled water/formic acid (19.5:80.2:0.3 v/v/v) as mobile phase. Tea samples were extracted by 75 % ethanol could get most high levels of EGCG and total catechins. The contents of caffeine and catechins have been measured in infusions of range of green tea, oolong tea, black tea and pu-erh tea products sold and consumed in the China, Japan, and Taiwan. When Twenty-nine commercial tea samples extracted by boiling water or 75 % ethanol and were analyzed by the HPLC method, the levels of EGCG and total catechins in teas were in the order: green tea (ripe leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea in substance. The contents of caffeine and catechins also have been measured in fresh tea leaves and tea flowers from Ping-Lin or Tai-Dong tea improvement mill. The old leaves contain less caffeine but more EGCG and total catechins than young leaves. The tea flowers contain very low caffeine but high levels of EGCG and total catechins as well as the old leaves. To comparison of caffeine and catechins in the same tea but manufactured different level of fermentation, the fresh leaves contain low levels of caffeine, EGCG and catechins. The level of caffeine in different manufactured tea was in the order: black tea > oolong tea > green tea > fresh leaf, but the levels of EGCG and total catechins were in the order: green tea > oolong > fresh leaf > black.
In addition, we used six commercial tea and three parts of one tea tree species (including old, young leaves and flowers) samples extract to test the biological functions including antioxidative, anti-inflammatory activity, and induction of apoptosis. The pu-erh tea crude extracts protected the plasmid DNA damage by Fenton reaction as well as the control at a concentration 100 g/mL. The antioxidateve activity of tea crude extracts was in the order: pu-erh tea > green tea > black tea > oolong tea. All crude extracts were found to reduce NO generation in a concentration 50 g/mL. The pu-erh tea and black tea extracts have very strong activity to inhibition of LPS-induced NO generation. Analyzed the activity of crude extracts on induction of apoptosis by DNA fragmentation ladder and flow cytometry, the ability of tea extracts to induce HL-60 cells apoptosis were in the order: green tea > oolong > black tea > pu-erh tea. And the ability of fresh old leaf extracts to induce HL-60 cells apoptosis were as well as the oolong tea extracts. To deserve to be mentioned, all crude extracts extracted by 75 % ethanol have more strong biological functions than extracted by boiling water.

Title page…………………………………………………………………Ⅰ
Signature of thesis committee…………………………………………Ⅱ
A power of attorney………………………………………………………Ⅲ
Acknowledgments……………………………………………………………Ⅳ
Lists of contents…………………………………………………………Ⅴ
Lists of tables……………………………………………………………Ⅵ
Lists of figures………………………………………………………………………Ⅶ
Abstract in Chinese………………………………………………………1
Introduction in Chinese…………………………………………………2
Abstract……………………………………………………………………11
Introduction………………………………………………………………13
Materials and Methods……………………………………………………16
Results………………………………………………………………………21
Discussion…………………………………………………………………27
References…………………………………………………………………30
Figures and Tables………………………………………………………34

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