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研究生:陳宜嫺
研究生(外文):I-Hsien
論文名稱:蔓越莓汁酚類化合物之安定性與乙酸乙酯萃取物及其區分物對於因藍光受損ARPE-19細胞之修復功效
論文名稱(外文):The phenolics stability in cranberry juice and the repairment of the ethyl acetate extract and its separated fractions on damaged ARPE-19 cells irradiated by blue light
指導教授:王進崑王進崑引用關係陳肅霖陳肅霖引用關係
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:營養學研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:74
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蔓越莓具有高抗氧化性及生物活性,包括抗癌、預防心血管疾病、預防泌尿道感染、預防胃潰瘍等功效,頗受消費者矚目。蔓越莓含有豐富的酚類化合物,本研究針對蔓越莓中酚類化合物之儲存安定性及其對黃斑部病變之影響進行探討。結果顯示,蔓越莓中的酚類化合物含量會隨儲存時間而下降,其中以游離態quercetin之安定性最佳,總類黃酮及前花青素則其次,在總酚類化合物之安定性最差。而在其乙酸乙酯萃取物中,以含縮合單寧區分物之抗氧化能力最好。在細胞實驗中,利用藍光(波長為450nm)照射誘使人類視網膜色素上皮細胞株(ARPE-19細胞)受損的試驗模式可知,蔓越莓中之縮合單寧區分物具有修復人類眼球上皮細胞損害的功效,而又以含寡縮合單寧區分物(oligomeric condensed tannin)之效果較佳。由本研究結果可知,蔓越莓可能具有清除自由基以及修復人類眼球上皮細胞損害的功效,進而預防因自由基所誘發之黃斑部病變之發生。

Cranberry shows high antioxidation and biological activities, including anticancer, prevention of cardiovascular disease, prevention of urinary tract infection, prevention of gastric ulcer and so on. Cranberry contains abundant phenolics. Thus this study was focused on the storage stability of phenolics in cranberry and its effect on age-related macular degeneration(AMD). The results showed that the phenolics in cranberry would reduced. The free quercetin showed the best stability. And the flavonoids and proanthocyanidins were the second. The total phenolics showed the worst stability. In the ethyl acetate extract of cranberry, the condensed tannin containing fraction showed the best antioxidant activity. In the cell experiments, the model of damaged ARPE-19 cells irradiated by blue light (450nm)showed that the condensed tannin containing fraction in cranberry could repair the damaged ARPE-19 cells irradiated by blue-light. Moreover, the oligomeric condensed tannin containing fraction showed the best efficiency. In conclusion, this study clearly showed that cranberry could scavenge free radicals and cause good efficiency on repairing ARPE-19 cells. This probably showed that cranberry could prevent the age-related
macular degeneration causing by free radicals.


目 錄
頁次
表次……………………………………………………………………………………I
圖次…………………………………………………………………………………II
中文摘要……………………………………………………………………………IV
英文摘要……………………………………………………………………………V
壹. 前言………………………………………………………………………………1
貳. 文獻探討……………………………………………………………2
一、 蔓越莓………………………………………………………………………2
(一) 蔓越莓的簡介………………………………………………… 2
(二) 蔓越莓的組成份………………………………………………2
(三) 蔓越莓的生理功效……………………………………………8
二、 酚類化合物………………………………………………………10
(一) 酚類化合物的定義與分類……………………………………10
(二) 酚類化合物的生理活性………………………………………18
(三) 酚類化合物與抗氧化…………………………………………18
(四) 酚類化合物與抗發炎…………………………………………18
三、 前花青素…………………………………………………………19
(一) 結構……………………………………………………………19
(二) 生理活性………………………………………………………20
1. 抗氧化………………………………………………………20
2. 抗癌…………………………………………………………20
3. 抑菌…………………………………………………………21
4. 預防白內障…………………………………………………21
四、 人類眼球視網膜色素上皮細胞…………………………………21
參. 研究目的………………………………………………………………… 24
肆. 實驗流程………………………………………………………………… 25
伍. 材料方法………………………………………………………………… 26
一、 試驗材料…………………………………………………………26
(一) 蔓越莓與蔓越莓濃縮原汁…………………………………26
(二) 蔓越莓汁儲存條件…………………………………………26
(三) 蔓越莓汁乙酸乙酯萃取物製備……………………………26
(四) 乙酸乙酯萃取物之區分……………………………………26
(五) 含縮合單寧區分物之再區分………………………………27
二、 試驗用藥…………………………………………………………27
三、 試驗儀器…………………………………………………………29
四、 試驗方法…………………………………………………………29
(一) 酚類化合物之含量分析……………………………………29
1. 總酚類化合物含量…………………………………………29
2. 總類黃酮化合物含量………………………………………30
3. 前花青素含量測定…………………………………………30
4. quercetin(free)含量的測定…………………………………30
5. 縮合單寧含量的測定………………………………………31
6. 單寧之確認…………………………………………………31
(二) 抗氧化活性試驗……………………………………………31
1. 總抗氧化能力………………………………………………31
2. 還原力之測定………………………………………………31
3. DPPH自由基清除能力測定…………………………………32
(三) 細胞培養……………………………………………………32
(四) 薄層層析法…………………………………………………34
(五) 酚類化合物之高效能液相層析……………………………34
陸. 統計分析……………………………………………………………………… 36
柒. 結果與討論……………………………………………………………… 37
一、 蔓越莓中酚類化合物之安定性試驗…………………………… 37
二、 蔓越莓濃縮汁中酚類化合物含量分析………………………… 42
三、 蔓越莓汁乙酸乙酯萃取物及其區分物中酚類化合物之分析………………………………………………………………………… 42
(一) 蔓越莓汁乙酸乙酯萃取物之製備…………………………42
(二) 乙酸乙酯萃取物之區分……………………………………42
(三) 酚類化合物之分析…………………………………………42
(四) 酚類化合物之HPLC分析…………………………………45
四、 蔓越莓汁乙酸乙酯萃取物及其區分物之生物活性探討…………………………………………………………………………45
(一) 抗氧化活性試驗……………………………………………45
(二) 蔓越莓汁乙酸乙酯萃取物對ARPE-19細胞增生之
影響…………………………………………………………52
(三) 蔓越莓乙酸乙酯萃取物之再區分物對受損ARPE-19
五、 細胞增生之影響……………………………………………57
(一) 含縮合單寧區分物之再區分………………………………57
(二) 含縮合單寧區分物之再區分物成份分析…………………59
(三) 含縮合單寧區分物之再區分物對ARPE-19細胞增
生之影響………………………………………………………62
捌. 結論…………………………………………………………………………… 65
玖. 參考文獻……………………………………………………………………… 66


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