臺灣博碩士論文加值系統

摘要
本研究主要探討十種花草茶原料 (馬鞭草、香蜂草、歐蓍草、牛膝草、鼠尾草、蕁麻葉、忘憂草、柚子葉、檸檬葉、天仙果) 的熱水萃取物之抗氧化能力比較分析。首先測量總多酚含量和類黃酮總量來評估熱水萃物的整體抗氧化成分含量，接著以體外抗氧化實驗 (DPPH 和 ABTS 自由基清除能力、還原力、亞鐵離子螯合力和總抗氧化力) 比較十種熱水萃取物的抗氧化能力，並進一步用 HPLC 分析抗氧化成分。
在總多酚含量方面，香蜂草總酚量 (388.51 mg/g) 遠高於其他熱水萃取物，其次為牛膝草 (164.85 mg/g) 和鼠尾草 (149.34 mg/g) 而蕁麻葉最低 (20.52 mg/g)，類黃酮含量則是以忘憂草 (41.99 mg/g)、馬鞭草 (37.61 mg/g) 和牛膝草 (36.47 mg/g) 最高，而檸檬葉 (9.59 mg/g) 最低。在體外抗氧化能力方面，香蜂草、牛膝草與鼠尾草有最佳的 DPPH 自由基清除能力與還原力。亞鐵離子螯合力方面則是以蕁麻葉、歐蓍草和馬鞭草有最佳效果。ABTS自由清除能力則是香蜂草、牛膝草和忘憂草最佳。總抗氧化力方面，香蜂草、忘憂草、馬鞭草效果最強。 總多酚含量與四種試驗有較高的相關性 (DPPH 自由基清除能力、還原力、ABTS 自由基清除能力和總抗氧化力)。抑制酪胺酸酶活性方面，只有天仙果和柚子葉具抑制效果。抑制黃嘌呤氧化酶活性方面，香蜂草在濃度 0.5 mg/ml 時有最佳效果 (55.7%)。
HPLC 分析方面，選擇六種抗氧化效果較佳的花草茶熱水萃物 (馬鞭草、香蜂草、歐蓍草、牛膝草、鼠尾草和忘憂草) 進行分析，定量出三種化合物 (咖啡酸、迷迭香酸和木犀草素)，總量介於 0.83-63.66 mg/g 之間。

關鍵字：抗氧化、HPLC、花草茶、萃取物。

Abstract
This study focused on antioxidant activity of hot water extract for ten kinds of herbal tea (Verbena officinalis, Melissa officinalis, Achillea millefolium, Hyssopus officinalis, Salvia officinalis, Urtica dioica, Hemerocallis fulva, Citrus maxima, Citrus limon, Ficus formosana ). First, total polyphenol content and total flavonoids were measured to evaluate the antioxidant components of hot water extracted material. Followed by in vitro experiments including DPPH and ABTS radicals scavenging activity, reducing and chelating power, and total antioxidant activity were selected to investigate antioxidant capacities. Furthermore, HPLC was used to identify antioxidant compound..
The amount of total polyphenol content in Melissa officinalis (388.51 mg/g) was much higher than other hotwater extracts, followed by Hyssopus officinalis (164.85 mg/g) and Salvia officinalis (149.34 mg/g). Urtica dioica was the lowest (20.52 mg/g). The total flavonoid contents were abundant in Hemerocallis fulva (41.99 mg/g), Verbena officinalis (37.61mg/g) and Hyssopus officinalis (36.47 mg/g). Citrus limon was the lowest (9.59 mg/g). In terms of antioxidant capacity, Melissa officinalis, Hyssopus officinalis and Salvia officinalis had the highest DPPH radical scavenging capacity and reducing power. For ABTS radical scavenging capacity, Melissa officinalis, Hyssopus officinalis and Hemerocallis fulva had the highest capacity. For chelating effect, Urtica dioica, Achillea millefolium and Verbena officinalis had the highest effect. For total antioxidant capacity, Melissa officinalis, Hemerocallis fulva and Verbena officinalis had the best capacity. Total polyphenol contents correlated well with four antioxidant activities (DPPH radical scavenging capacity, reducing power, ABTS radical scavenging capacity, and total antioxidant capacity). Only Citrus maxima and Ficus formosana had the inhibitory effect of the tyrosinase activity. The Melissa officinalis had the best results at 0.5 mg/ml (55.7%) against xanthine oxidase activity.
The six kinds of hot water extracts of herbal tea (Verbena officinalis, Melissa officinalis, Achillea millefolium, Hyssopus officinalis, Salvia officinalis and Hemerocallis fulva) which have better antioxidant activities were selected for HPLC analysis. Three compounds (caffeic acid, rosmarinic acid and luteolin) were found to be present in the extract. The total amount of these compounds in samples was 0.83-63.66 mg/g.

Keywords: antioxidative, HPLC, herbal tea, extract

目錄
壹、 前言 1
貳、 文獻回顧 2
一、 十種花草茶植物介紹 2
(一) 馬鞭草 2
(二) 香蜂草 2
(三) 歐蓍草 3
(四) 牛膝草 3
(五) 鼠尾草 4
(六) 蕁麻葉 4
(七) 忘憂草 4
(八) 柚子葉 5
(九) 檸檬葉 5
(十) 天仙果 5
二、 活性氧與自由基 6
(一) 活性氧與自由基的形成 6
(二) 氧化壓力所造成的危害 6
(三) 常見的自由基與活性氧 7
三、 活性氧的防禦 8
(一) 抗氧化防禦機制 8
(二) 抗氧化劑作用機制 9
(三) 抗氧化酵素 10
(四) DNA 修復 12
四、 高效能液相層析法 (High performance liquid chromatography, HPLC) 13
參、 實驗設計 14
肆、 材料與方法 15
一、 實驗材料 15
(一) 樣品萃取之材料 15
(二) 抗氧化成分定量分析之材料 15
(三) 抗氧化能力測試之材料 15
(四) HPLC之材料 17
二、 實驗方法 17
(一) 樣品萃取 17
(二) 抗氧化成分定量分析 17
(三) 抗氧化能力測試 18
(四) HPLC 21
(五) 統計分析 22
伍、 結果與討論 23
一、 花草茶熱水萃取物萃取率 23
二、 總多酚含量 23
三、 總類黃酮含量 23
四、 DPPH 自由基清除能力 24
五、 亞鐵離子螯合能力 24
六、 還原力 25
七、 ABTS 自由基清除能力 25
八、 總抗氧化力 26
九、 總多酚含量與不同抗氧化活性的相關性 26
十、 DPPH 自由基清除能力與其它抗氧化能力的相關性 27
十一、 酪胺酸酶抑制活性測定 27
十二、 黃嘌呤氧化酶抑制活性 28
十三、 HPLC 分析 28
陸、 結論 30
柒、 參考文獻 31
捌、 圖 38
玖、 表 61
壹拾、 附錄 65

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