臺灣博碩士論文加值系統

摘要
本研究主要是探討綠藻水萃物之抗氧化性質，以更進一步了解綠藻之機能性。實驗藉由試驗設計方式針對加熱溫度和時間兩個因素進行水萃取，以體外化學分析測定抗氧化能力，最後以反應曲面進行統計分析，決定具有較佳抗氧化性之最適萃取條件。結果顯示，綠藻水萃物1-14組於濃度10 mg/mL時皆可清除DPPH自由基達80%以上，以反應曲面進行統計分析，較高的反應速率常數k值分別出現在較低萃取溫度40-50℃(0.3 mL/mg)之間和100℃(0.3 mL/mg)兩個區域，時間的影響上則是低溫不顯著，高溫100℃短時間較佳。螯合亞鐵能力方面皆隨水萃物濃度增加而增加，反應數率常數k值隨著萃取溫度的上升而增加，在100℃時最高(0.14 mL/mg)而時間則以短時間約1小時可獲較大k值。還原力皆隨萃取物濃度升高而增加，結果分別在100℃、1小時和40℃、5小時可獲得較高之k值0.135 mL/mg。不同萃取條件所得總酚類含量在80-100℃之間及萃取時間約4小時，可獲得總酚量約2.95 mg/g。水萃取率則隨加熱溫度的上升可獲得較高的萃取率約18.0%，而時間上則是在3小時以內具有較高的萃取率。綜合以上分析結果，決定綠藻較佳抗氧化效果之萃取條件為高溫100℃和1小時為最適條件組合。
另外在酚類化合物分析上，比對層析圖各個尖峰面積大小與抗氧化間之相關性，以Pearson Correlation分析結果顯示層析沖提時間在7.51分鐘之尖峰面積與DPPH清除能力有顯著正相關r = 0.733 (p＜0.01)，而在此沖提時間下之紫外光譜圖於219和274 nm有最大吸收波長，推測可能具有酚類化合物。綠藻水萃物以100℃和1小時萃取之萃取物，單糖組成主要以葡萄糖(52.37±2.82)和核糖(25.89±0.89)為主(以莫耳百分率表示)，約佔77%左右。其分子量分佈可分成兩種分子量族群F1(24.97±0.6 kDa)和F2 (0.95±0.09 kDa)。

Abstract
This study was to investigate the antioxidant activities of Chlorella water extracts with various extraction conditions by the central composite design, where the independent variables were extraction temperature and time. The data were statistically analyzed with response surface method to find the optimal extraction condition for a better antioxidant activity. The results of antioxidation measurements indicated that all extracts showed a good DPPH-radical scavenging ability above 80% at 10 mg/mL. A greater scavenging rate constant presented at about 4050oC and 100oC according to the response surface analysis. The higher the temperature, the shorter was the period. The Fe2+-chelating ability increased with increasing the extract concentration. The rate constant k increased with the elevated temperature and maximized at 100oC for 1 hr (0.14 mL/mg). The Fe2+-reducing power rose with the increased extract concentration and maximized up to 0.135 mL/mg at 100oC for 1 hr and 40oC for 5 hr. The total phenol contents of the extracts at 80-100oC for about 4 hr were about 2.95 mg/g. The yield of water extracts increased to 18.0% with increasing temperature and for within 3 hr. Generally, the optimal extraction condition for Chlorella extracts with a higher antioxidant activity was 100oC and 1 hr.
The results of HPLC-PDA analysis indicate that the 75% ethanol soluble of Chlorella water extracts exhibited a signal at 7.51 min and absorbance peaks at 219 and 274 nm, point to the possible presence of phenolic compounds. The monosaccharide compositions of Chlorella water extract at 100℃ for 1 hr were mainly composed of glucose (52.37±2.82%) and ribose (25.89±0.89%), totally for about 77%, and having a bimolecular distribution of F1 (24.97±0.6 kDa) and F2 (0.95±0.09 kDa).

目錄
中文摘要……………………………………………………………………..Ⅰ
英文摘要……………………………………………………………………..Ⅱ
目錄…………………………………………………………………………..Ⅲ
表目錄………………………………………………………………………..Ⅶ
圖目錄………………………………………………………………………..Ⅸ
壹、前言………………………………………………………………………1
貳、文獻回顧………………………………………………………….……...2
一. 綠藻之分類………………………………………………….………2
二. 綠藻之生活習性與培養方式………………………………..………4
三. 綠藻之成分組成…………………………………………..…………6
1.營養成分組成…………………………………………...……………6
2.多醣及細胞壁組成分組成………………………………………….10
四. 綠藻之保健功效研究………………………………………….…....11
1. 提升抗氧化能力……………………………………………….…....11
2. 脂質代謝影響………………………………………………….……11
3. 綠藻對免疫系統的功能促進………………………………….…....13
4.幫助體內排除戴奧辛毒物………………………………………….14
5.綠藻對高血壓患者的影響………………………………………….14
五. 綠藻在工業之應用……………………………………………….....15
六. 抗氧化劑作用機制……….…………………………………….……15
6.1抗氧化劑之作用機制……………………………………………….15
6.2 抗氧化劑之介紹……………………………………………………16
6.2.1人工合成抗氧化劑……………………………………………...16
6.2.2天然抗氧化劑…………………………………………………...18
七.實驗設計…………………………………………………………...….27
叁、研究目的…………………………………………………………………29
肆、材料與方法………………………………………………………………30
一、實驗流程…………………………………………………………...30
二、樣品製備……………………………………………………………31
三、化學藥品…………………………………………………………….31
四、綠藻水萃取物之萃取方式………………………………………….32
五、化學分析…………………………………………………………….32
六、綠藻水萃物抗氧化分析…………………………………………….35
6.1 捕捉1,1-二苯基-2-苦味………………………………………..35
6.2 螯合亞鐵離子之測定…………………………………………..36
6.3 還原力測定……………………………………………………..36
6.4 酚類化合物測定(HPLC-PDA)…………………………………38
6.5萃取物對清除自由基效力及還原力反應模式 ...……………………………………………………………38
七、分子性質與單糖組成測定………………………………………....40
7.1分子性質之測定……………………….……………………….40
7.2 單醣組成分析………………………….……………………….40
八、統計分析……………………………………………………………42
伍、結果與討論………………………………………………………………43
1、萃取時間與溫度對綠藻水萃物萃取率之影響……………………….43
2、 萃取時間與溫度對綠藻水萃物總酚類化合物(total phenolic compound)含量之影響…………………………………………………………….43
3、萃取條件對綠藻萃取物清除DPPH自由基之影響………………….46
4、不同萃取條件之萃取物之螯合亞鐵離子能力……………………….49
5、不同萃取條件綠藻萃取物之還原力………………………………….49
6、酚類化合物分析(HPLC-PDA)…………………………………………61
7、綠藻萃取物分子性質………………………………………………….65
陸、結論………………………………………………………………………69
柒、參考文獻…………………………………………………………………70

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