臺灣博碩士論文加值系統

本研究以市售之烏龍茶為研究對象，探討其茶湯中各成分苦澀味成分如兒茶素類化合物、咖啡因等。甘味成分如胺基酸及茶胺酸及香氣成分等的分析。並嘗試以不同的水溫度（70、80及90℃）和時間（1、2及3 分鐘）萃取烏龍茶，探討其對苦味、澀味、香味及甘味成分之對溶出影響，並分析各種萃取條件所得茶湯中兒茶素類化合物、咖啡因、總胺基酸、茶胺酸及茶色等成分之變化，再依此實驗之結果選擇較佳沖泡條件，以求得茶湯中較多茶多酚含量、較多甘味成分之溶出、較多香氣成分含量及最少咖啡因含量。並以90℃水溫進行烏龍茶之不同沖泡次數對茶湯中成分之探討，其沖泡條件分別為第一泡至第五泡之沖泡時間分別為1、1.5、2、2.5及3分鐘，從第六泡至第十泡沖泡時間都維持3分鐘。同時並自茶湯中抽取香氣成分以GC-MS進行茶湯之香氣成分之定性及定量，並分析各泡茶湯中兒茶素類、咖啡因、總胺基酸及茶胺酸之含量變化可供作泡健康茶及製備飲料茶的依據。
茲將本試驗之結果歸納如下：
1、 不同萃取溫度與時間之茶湯，依所建立之分析結果顯示茶湯中各成分之含量都隨著萃取溫度增加及萃取時間加長而增加。茶湯中之總兒茶素類含量於70℃時1~3分鐘之萃取量為4.35∼7.31 mg/g、80℃為5.03∼7.92 mg/g、90℃為6.04∼10.27 mg/g。總胺基酸含量70℃為0.79∼1.20 mg/g、80℃為0.84∼2.86 mg/g、90℃為1.17∼3.15 mg/g。而咖啡因之含量70℃為0.41∼0.94 mg/g、80℃為0.60∼1.13 mg/g、90℃為0.80∼1.70 mg/g。
2、 烏龍茶在90℃下連續沖泡十次，其各茶湯中總兒茶素之含量分別為5.88、9.84、12.13、10.74、12.31、12.59、12.65、11.73、10.19及9.35 mg/g，累積10次茶湯中之兒茶素化合物量為茶葉中所含兒茶素的88%。
3、 甘味成分的胺基酸在第1泡及第2泡茶湯並不高，僅有1.0~1.2 mg/g，第3~5泡茶湯中分別含有約2.9、3.5、3.0 mg/g左右，第6泡以後到第10泡則仍維持在1.0 mg/g 左右，顯示第3~5泡之茶湯最具甘味。
4、 茶湯中咖啡因含量以第一次沖泡之茶湯量最少，約為0.7 mg/g，隨後2~10次茶湯中之量雖有增減，但均維持在1.5~2.0 mg/g上下。
5、 烏龍茶茶湯之香氣成分GC-MS分析結果共鑑定出36個化合物，如phenylethyl alcohol、methyl jasmonate及linaool及indole等。其主要香氣成分為具類葡萄柚香及溫和花香之indole。在第2泡時有最大之香氣含量約為2898 μg/mL，隨沖泡次數進行至第5泡其含量減為1296 μg/mL，至第10泡其含量僅剩186.5 μg/mL。
6、 茶湯中之DPPH捕捉自由基能力與茶湯中之兒茶素量呈正的相關。
7、 建立茶湯中各成分之分析方法以作為模式評估之參考依據。

The study focus on commercial oolong tea to investigate the content of its bitter-stringent components such as catechin and caffeine, the sweet components such as amino acids and theanine, and the fragrance components in tea. We attempted to infuse the tea leaves in water at different temperatures (70, 80 and 90℃) and infusion time (1, 2 and 3 min) to extract the oolong tea and investigated the effects of brewing conditions on dissolution of bitter, stringent, fragrant and sweet components in the tea infusions. Furthermore, the effects of various extraction conditions on content variations of tea infusions regarding catechin, caffeine, total amino acids, theanine, and tea color were examined. Based on the experimental results, the better brewing conditions were selected in order to obtain higher catechin content, greater sweet components, greater fragrance, and least caffeine content. We also examined number of oolong tea brewing using 90℃ water on the components of tea infusions. The brewing conditions for the 1st to 5th brewing were 1, 1.5, 2, 2.5 and 3 min, respectively; and from 6th to 10th brewing, 3 min intervals were applied. The fragrant components from the tea infusions were extracted and analyzed qualitatively and quantitatively using GC-MS. The catechin, caffeine, total amino acids, and theanine content variations could serve as criteria for preparing most healthful tea and beverage teas.

The results of the study are summarized below:
1. Based on the results of analysis, various components of the tea infusions obtained using different brewing temperatures and times tend to increase with brewing times. The total catechin content for teas brewed at 70℃ for 1~3 min was 4.35 to 7.31 mg/g; those brewed at 80℃ was 5.03 to 7.92 mg/g; and those brewed at 90℃ was 6.04 to 10.27 mg/g. The total amino acids content for teas brewed at 70℃ was 0.79 to 1.20 mg/g; those brewed at 80℃ was 1.17 to 3.15 mg/g; and those brewed at 90℃ was 1.17 to 3.15 mg/g. For caffeine content, teas brewed at 70℃ had 0.41 to 0.94 mg/L; those at 80℃had 0.60 gto 1.13 mg/g; and those at 90℃ had 0.80 to 1.70 mg/g.
2. Ten consecutive brewing at 90℃ gave tea liquid catechin content of 5.88, 9.84, 12.13, 10.74, 12.31, 12.59, 11.73, 10.19 and 9.35 mg/g, respectively. The cumulative catechin content of the 10 brewing accounted for 88% of all catechin in the tea leaves.
3. The sweet component, amino acids were relatively low in the 1st and 2nd infusions, containing 1.0 to 1.2 mg/g, respectively. In the 3rd to 5th infusion, they reached 2.9, 3.5 and 3.0 mg/g, respectively. In the 6th to 10th infusions, there were still ca. 1.0 mg/g content. Thus the 3rd to 5th infusions were particularly sweet.
4. There was the least caffeine content in the 1st infusion of the oolong tea, ca. 0.7 mg/g; in the 2nd to 10th infusions, caffeine content varied, but maintained a level of 1.5 to 2.0 mg/g.
5. Thrity-six compounds were identified by GC-MS for the fragrance of the tea infusions. These included phenylethyl alcohol, metyl jasmonate, linalool and indole etc. The main component was indole with fragrance akin to grapefruit and mild flora scent. The fragrances were at the maximum in the 2nd infusion reaching a content of 2898 μg/mL. These decreased to 1296 μg/mL in the 5th infusion and 186.5 μg/mL in the 10th.
6. In the tea infusions, the free radical scavenging capacity determined by DPPH assay showed a positive correlation with the catechin content.
7. In the study, we established the methods of analysis various components in tea infusions which could be used as a reference for model evaluations.

目錄
中文摘要 I
英文摘要 II
目錄 VI
圖目錄 VIII
表目錄 XI
壹、前言 1
貳、文獻整理 2
一、茶的成分 2
(一) 多酚類化合物 2
(二) 植物鹼 3
(三) 胺基酸 4
(四) 維生素 5
(五) 無機元素 5
(六) 揮發性香氣成分 6
二、兒茶素類之機能性介紹 6
(一) 抗氧化活性 6
(二) 抗菌性 7
(三) 抑制血壓上升效果 8
(四) 預防口腔疾病效果 8
(五) 兒茶素對血醣上升之抑制效果 9
三、茶葉中兒茶素類化合物、咖啡因及茶胺酸之分析 9
(一) 薄層層析法（TLC） 9
(二) 高效能液相層析法（HPLC） 10
(三) 高效能毛細管電泳分析方法（HPCE） 11
四、不同泡茶條件對茶湯中生理活性物質的影響 12
五、茶葉中揮發性香氣成分之分析 13
(一) 茶葉香氣成分之萃取法 13
1. 頂空間萃取法 13
2. 吹掃捕集法 14
3. 超臨界流體萃取 14
4. 固相微萃取 15
(二) 茶葉香氣成分之鑑定方法 15
1. 滯留時間指數 16
2. Mass –spectra library date 16
3. 標準品比對法 16
(三) 不同茶葉中揮發性香氣成分 17
1. 紅茶 17
2. 綠茶 18
3. 烏龍茶 18
六、抗氧化活性之探討 19
(一) 自由基定義 19
(二) 活性氧定義 19
(三) 抗氧化劑之作用與原理 20
1. 自由基終止型 20
2. 還原劑或氧原子清除劑者 21
3. 螫合劑型 21
4. 單重氧抑制劑 21

參、材料與方法 22
一、實驗材料 22
(一) 樣品 22
(二) 標準品 22
(三) 化學藥品 22
(四) 層析管柱 23
(五) 儀器 23
二、實驗方法 24
(一) 茶葉中兒茶素類和咖啡因之萃取定量 24
(二) 茶葉之沖泡與茶湯之色澤分析 25
(三) 以HPLC法進行茶湯中成分之分析 25
(四) 以毛細管電泳進行茶湯中茶胺酸之分析 26
(五) 茶湯中揮發性成分分析 28
(六) 茶湯中總胺基酸含量測定 29
(七) 以不同次數沖泡烏龍茶茶葉之方法 31
(八) 茶湯之DPPH自由基清除能力之測定 31
肆、結果與討論 33
一、茶葉及茶湯中各成分之分析方法的確立 32
(一) 咖啡因與兒茶素類之分析 32
(二) 咖啡因與兒茶素類之檢量製作 32
(三) 胺基酸之分析 33
(四) 茶胺酸之分析 33
二、烏龍茶茶葉中咖啡因與兒茶素之總量鑑定及分析 34
三、不同萃取溫度與時間對烏龍茶茶湯成分的影響 35
(一) 萃取時間對兒茶素類和咖啡因之含量變化的影響 36
(二) 萃取溫度的影響 36
(三) 胺基酸之含量 37
(四) 萃取時間對茶湯中茶胺酸之含量的影響 38
(五)萃取溫度對胺基酸的影響 38 (六) 茶色 38
四、烏龍茶於不同的沖泡次數對茶湯成分之影響 39
(一) 兒茶素類和咖啡因之含量 39
(二) 總胺基酸與茶胺酸之含量 40
(三) 茶色 40
五、DPPH自由基清除能力 40
六、烏龍茶之揮發性香氣成分組成 41
(一) 香氣成分之描述 41
(二) 不同沖泡次數之茶湯中香氣成分之變化 41
伍、結論 41
陸、參考文獻 43







圖目錄
圖一、茶葉之多酚類化合物之化學結構 54
圖二、甲基黃嘌呤化合物之化學結構 55
圖三、茶葉中茶胺酸之化學結構 56
圖四、茶葉中兒茶素抑制高血壓自發症鼠血壓上升的作用 57
圖五、茶葉中兒茶素對人體血糖值上升的抑制效果 58
圖六、烏龍茶茶葉香氣之萃取流程圖 59
圖七、以HPLC製作咖啡因和兒茶素衍生物之檢量線 60
圖八、以分光光度計製作總胺基酸之檢量線 61
圖九、以毛細管電泳製作茶胺酸之檢量線 62
圖十、烏龍茶逾70浸泡1分鐘茶湯之液相層析圖 63
圖十一、烏龍茶於70℃浸泡2分鐘茶湯之液相層析圖 64
圖十二、烏龍茶於70℃浸泡3分鐘茶湯之液相層析圖 65
圖十三、烏龍茶於80℃浸泡1分鐘茶湯之液相層析圖 66
圖十四、烏龍茶於80℃浸泡2分鐘茶湯之液相層析圖 67
圖十五、烏龍茶於80℃浸泡3分鐘茶湯之液相層析圖 68
圖十六、烏龍茶於90℃浸泡1分鐘茶湯之液相層析圖 69
圖十七、烏龍茶於90℃浸泡2分鐘茶湯之液相層析圖 70
圖十八、烏龍茶於90℃浸泡3分鐘茶湯之液相層析圖 71
圖十九、烏龍茶於70 ℃萃取1、2及3分鐘之兒茶素及咖啡因含量 72
圖二十、烏龍茶於80 ℃萃取1、2及3分鐘之兒茶素及咖啡因含量 73
圖二十一、烏龍茶於90 ℃萃取1、2及3分鐘之兒茶素及咖啡因含量
74
圖二十二、不同的萃取條件對烏龍茶湯中之兒茶素含量之影響 75
圖二十三、不同的萃取條件對烏龍茶湯中之咖啡因含量之影響 76
圖二十五、以茶於70℃浸泡1分鐘之烏龍茶湯於毛細管電泳圖 77
圖二十六、以茶於70℃浸泡2分鐘之烏龍茶湯於毛細管電泳圖 78
圖二十七、以茶於70℃浸泡3分鐘之烏龍茶湯於毛細管電泳圖 79
圖二十八、以茶於80℃浸泡1分鐘之烏龍茶湯於毛細管電泳圖 80
圖二十九、以茶於80℃浸泡2分鐘之烏龍茶湯於毛細管電泳圖 81
圖三十、以茶於80℃浸泡3分鐘之烏龍茶湯於毛細管電泳圖 82
圖三十一、以茶於90℃浸泡1分鐘之烏龍茶湯於毛細管電泳圖 83
圖三十二、以茶於90℃浸泡2分鐘之烏龍茶湯於毛細管電泳圖 84
圖三十三、以茶於90℃浸泡3分鐘之烏龍茶湯於毛細管電泳圖 85
圖三十四、第1泡之茶湯之烏龍茶湯之液相層析圖 86
圖三十五、第2泡之茶湯之烏龍茶湯之液相層析圖 87
圖三十六、第3泡之茶湯之烏龍茶湯之液相層析圖 88
圖三十七、第4泡之茶湯之烏龍茶湯之液相層析圖 89
圖三十八、第5泡之茶湯之烏龍茶湯之液相層析圖 90
圖三十九、第6泡之茶湯之烏龍茶湯之液相層析圖 91
圖四十、第7泡之茶湯之烏龍茶湯之液相層析圖 92
圖四十一、第8泡之茶湯之烏龍茶湯之液相層析圖 93
圖四十二、第9泡之茶湯之烏龍茶湯之液相層析圖 94
圖四十三、第10泡之茶湯之烏龍茶湯之液相層析圖 95
圖四十四、不同沖泡次數之烏龍茶湯中各兒茶素之含量 96
圖四十五、不同沖泡次數之烏龍茶湯中各兒茶素含量之影響 97圖四十六、沖泡時間對烏龍茶茶湯中兒茶素含量之影響 98
圖四十七、不同沖泡次數之烏龍茶湯中咖啡因含量之影響 99
圖四十八、沖泡時間對烏龍茶茶湯中咖啡因含量之影響 100
圖四十九、第1泡之烏龍茶湯之毛細管電泳圖 101
圖五十、第2泡之烏龍茶湯之毛細管電泳圖 102
圖五十一、第3泡之烏龍茶湯之毛細管電泳圖 103
圖五十二、第4泡之烏龍茶湯之毛細管電泳圖 104
圖五十三、第5泡之烏龍茶湯之毛細管電泳圖 105
圖五十四、第6泡之烏龍茶湯之毛細管電泳圖 106
圖五十五、第7泡之烏龍茶湯之毛細管電泳圖 107
圖五十六、第8泡之烏龍茶湯之毛細管電泳圖 108
圖五十七、第9泡之烏龍茶湯之毛細管電泳圖 109
圖五十八、第10泡之烏龍茶湯之毛細管電泳圖 110
圖五十九、不同沖泡次數之烏龍茶湯中總胺基酸與茶胺酸含量之影響 111
圖六十、不同沖泡次數之茶湯清除DPPH自由基之能力 112
圖六十一、兒茶素含量與DPPH自由基清除能力之相關性 113
圖六十二、第1泡之烏龍茶湯之GC-MS層析圖譜 114
圖六十三、第2泡之烏龍茶湯之GC-MS層析圖譜 115
圖六十四、第3泡之烏龍茶湯之GC-MS層析圖譜 116
圖六十五、第5泡之烏龍茶湯之GC-MS層析圖譜 117
圖六十六、第10泡之烏龍茶湯之GC-MS層析圖譜 118




表目錄
表一、茶葉和茶湯中的一般化學組成分 119
表二、兒茶素類對食物中毒細菌、乳酸菌、雙叉乳酸桿菌的抗菌活性 120
表三、茶葉多酚類對口腔齲菌之抑制情形 121
表四、烏龍茶以1：1（v/v）之乙醇水溶液萃取後之兒茶素類和咖啡因之含量 122
表五、不同萃取溫度與時間對烏龍茶胺基酸與茶胺酸含量之影響 123
表六、不同的萃取溫度與時間對茶湯色澤的影響 124
表七、不同的萃取次數對烏龍茶茶湯色澤的影響 125
表八、烏龍茶於不同沖泡次數之茶湯香氣成分組成 126
表九、烏龍茶於不同沖泡次數之茶湯香氣成分分類 127
附表一、不同的萃取溫度與時間對茶湯中兒茶素類成分的影響 128
附表二、不同沖泡次數對烏龍茶茶湯成分的影響 129

陸、參考文獻
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白禎芳（1993）沖泡條件對包種茶茶湯品質之影響。國立中興大學食品科學研究所碩士論文。
石正玲、朱延和（2001）化學。毛細管電泳於生物分子辦識及組合化學之應用。59：391-398。
李敏雄、陳漢龍、林基增、阮逸明（1984）烏龍茶之香氣成分及其品質。食品科學，11：126-133。
阮逸明（1991）茶葉可溶分及主要化學成分萃取之研究。臺灣茶葉研究彙報，10：89-108。
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