臺灣博碩士論文加值系統

酸柑茶為客家一種獨特飲品，具茶香及柑橘香並帶有些許澀感及苦感，其結合茶葉(Camellia sinensis)與虎頭柑(Citrus aurantium L. cv. Hutou Gan)以填塞、蒸煮、緊壓、乾燥與重覆等步驟製成，長久流傳酸柑茶具有止咳化痰之效。本篇研究之目的在於利用抗氧化、抗發炎及抗菌測試，並與其它茶類如紅茶、綠茶及普洱茶比較，評估酸柑茶之保健功效，了解酸柑茶作為保健飲品之潛力，賦與酸柑茶額外價值。

本實驗所用的抗氧化評估方法為DPPH自由基清除能力、ABTS自由基清除能力、FRAP鐵離子氧化還原能力測定、MICA亞鐵離子螯合能力、Folin-Ciocalteu reagent reaction 總酚含量及Flavonoid content assay總類黃酮含量測試；抗發炎以巨噬細胞NO產生量及存活性做評估；抗菌以抑菌圈紙錠擴散法及96孔盤微量稀釋測定法對金黃色葡球菌、大腸桿菌、沙門氏菌及李斯特菌作測試，並計算其最小抑菌濃度。

實驗結果顯示茶多酚及柑橘類黃酮與抗氧化有很大相關性，因此含量較低的酸柑茶於抗氧化機能性較一般茶類差，如綠茶、烏龍茶與紅茶，但抗發炎機能性及抗菌機能性卻較一般茶葉好。推測酸柑茶在熱加工過程中，可將兒茶素、柑橘類黃酮等聚合成未知大分子物質。酸柑茶於高濃度1000 μg/ml下，不僅促進小鼠巨噬細胞Raw 264.7的存活率，還具降低發炎物質一氧化氮的生成，並且可有效抑制對兒茶素EGCG較耐受性較高的革蘭氏陰性菌。

Tsuan-kan tea is a unique tea product of Hakka area in Taiwan. It combines Hutou Gan (Citrus aurantium L. cv. Hutou Gan) with tea and is produced by special manufacture procedures including steaming, baking and tying. Tsuan-kan teas are rich in aroma and with astringent and slightly bitter tastes. Antiinflammatory and reduce-phlegm formula of Tsuan-kan tea for the Hakka ancestors has been hand down for a long time. They claim that the tea can smooth redness symptoms of upper respiratory tract infection.

In this study, Anti-oxidation experiment was performed with electron and radical scavenging assay, DPPH free radical bleaching assay, ABTS free radical bleaching assay, Trolox equivalent antioxidant capacity assay, ferric reducing antioxidant power assay and metal ion chelating capacity assay. The anti-inflammatory effects of Tsuan-kan tea were investigated in vitro with LPS-induced RAW264.7 cells as the model. Antibacteria experiment with inhibition zone method was conducted by using common pathogens such as Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Salmonella typhimurium as indicators.

Result show that citrus flavonoids and polyphenols have great relevance to antioxidant, therefore lower content of tsuan-kan teas worse than the commercial tea (green tea, oolong tea and black tea), but the anti-inflammatory and antimicrobial activities is better than commercial tea. It suggested that the catechins or citrus flavonoids of Tsuan-kan tea can be polymerized to unknown substance during the thermal processing. Tsuan-kan teas (1000 μg/ml) is not only to promote the survival of Raw 264.7 cell in high concentration, but lower levels of inflammatory substances of nitric oxide, and can effectively suppress Gram-negative bacteria which are higher tolerance to catechin (EGCE).

謝致辭 I
摘要 II
Abstract III
目次 IV
圖目次 VIII
表目次 IX
縮寫對照表 X
壹、前言 1
貳、文獻回顧 2
一、酸柑茶由來 2
二、酸柑茶加工方法 3
三、酸柑茶泡茶方法及味道 4
三、酸柑茶於茶葉分類中之類別 5
四、酸柑茶茶湯基礎成分 5
(一) 茶葉 5
(二) 柑橘 6
五、酸柑茶茶湯機能性 8
(一) 抗氧化機能性 8
1. DPPH自由基清除能力 13
2. ABTS自由基清除能力 13
3. FRAP鐵離子氧化還原能力 13
4. MICA亞鐵離子螯合能力 14
5. Folin-Ciocalteu reagent reaction 總酚含量 14
6. Flavonoid content assay總類黃酮含量 14
(二) 抗發炎機能性 15
1. NO抗發炎測定實驗 16
(三) 抗菌機能性 16
肆、材料與方法 19
一、實驗材料 19
(一) 試驗用菌種 19
(二) 茶樣品 19
(三) 藥品與培養基 19
(四) 儀器 21
(五) 電腦套裝軟體 22
二、實驗架構 23
三、實驗方法 24
(一) 萃取方法 24
(二) 物理化學特性測定 24
1. pH值 24
2. 糖度 24
3. 導電度 24
4. 可溶性固體 24
5. 色澤分析 25
(三) 抗氧化能力測定 25
1. DPPH自由基清除能力測定法 25
2. ABTS自由基清除能力測定法 26
3. 鐵離子氧化還原能力測定(Ferric reducing antioxidant power, FRAP) 26
4. 金屬離子螯合能力(Metal ion chelating ability, MICA) 27
5. Folin-Ciocalteu reagent reaction 28
6. 總類黃酮測定法 28
(四) 抗發炎能力測定 29
1. 巨噬細胞NO測定法 29
2. MTT存活率測定法 30
(五) 抗菌能力測定 31
1. 抑菌圈紙錠擴散法(disc diffusion method) 31
2. 96孔盤微量稀釋測定法(96-microwell dilution method) 31
(六) 統計分析 32
伍、結果與討論 32
一、物理化學特性分析 33
(一) pH值 33
(二) 糖度 33
(三) 導電度 34
(四) 可溶性固體 34
(五) 色澤分析 35
(二) ABTS自由基清除能力結果 39
(三) 鐵離子氧化還原能力(FRAP)結果 40
(四) 金屬離子螯合能力(Metal ion chelating ability) 41
(五) 總酚含量 52
(六) 總類黃酮含量 56
(七) 抗氧化成分與抗氧化能力之相關性 60
三、抗發炎特性分析 63
四、抗菌特性分析 68
(一) 紙錠擴散法 68
(二) 96孔盤微量稀釋法之培養終點判定 71
(三) 96孔盤微量稀釋法 73
1. 抗金黃色葡萄球菌 73
2. 抗大腸桿菌 73
3. 抗沙門氏菌 73
4. 抗李斯特菌 74
陸、結論 81
柒、未來展望 82
捌、參考文獻 83

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