臺灣博碩士論文加值系統

為探討洛神葵及桑椹酒中花青素聚合物與其抗氧化能力之關係，本研究以UV/VIS測定花青素聚合物百分比，並利用清除DPPH自由基能力、還原力及Trolox當量之抗氧化能力(TEAC)三種抗氧化方法，分別就下列樣品加以測定。(1)洛神葵以不同溫度乾燥後，貯存於不同溫度下0~25週；(2)pH1及pH2之洛神葵花青素模擬系統於90℃下進行不同時間之加熱；(3)不同貯存時間之桑椹酒。結果顯示，以75℃乾燥之洛神葵，共呈色及聚合的現象較明顯，但其共呈色百分比卻隨著貯存溫度及時間之增加而下降。pH1及pH2之洛神葵花青素模擬系統，於90℃加熱後，發現兩者均無共呈色現象產生，且其花青素聚合物並不穩定，易受熱而裂解；在抗氧化能力分析方面，隨著加熱時間之增加，兩模擬系統之清除DPPH自由基能力上升，與經由膠過濾純化後收集之FractionⅡ之DPPH清除能力有相同變化趨勢，而隨著加熱時間增加，模擬系統的還原力下降與純化後收集之FractionⅢ有相同趨勢；但於TEAC測定上兩者間並無顯著差異。在不同貯存期之桑椹酒測定方面，貯存一年之桑椹酒其共呈色及聚合物百分比最高；在抗氧化能力方面，清除DPPH自由基能力隨著貯存期增長而增加，原因是來自於聚合物的增加；還原力方面，隨貯存期增長其花青素含量漸減，因而還原力逐漸下降；在TEAC方面，各貯存期的桑椹酒其TEAC皆已超出所能測到的值，且經膠體過濾後發現 FractionⅡ具有較強的TEAC。根據上述試驗結果，證明清除自由基能力可隨著聚合物增加而增強，因此我們推論花青素聚合物與抗氧化能力之間確實相關，而與DPPH清除能力呈現正相關。

In order to investigate the relationship between degree of polymerization and antioxidant capacity in anthocyanin of roselle and mulberry wine, UV/VIS was used to measure degree of polymerization of samples, and scavenging activity on α,α-β-picrylhydrazyl (DPPH) radical assay、ferric reducing ability of plasma (FRAP) assay and Trolox equivalent antioxidant capacity (TEAC) assay, were used to measure antioxidant capacity of the following samples. Ⅰ. roselles dried by different temperature, and stored 0~25weeks under different storage temperature; Ⅱ. roselle anthocyanin model systems (pH1 and pH2) heated at 90℃with different time; Ⅲ. mulberry wine with different storage time. Results showed that, for the roselle sample dried under 75℃, the copigmentation and polymerization appeared obviously, but the percentage of copigmentation was decreased by increasing storage temperature and time. Copigment was not evident in both pH1 and pH2 roselle anthocyanin model system after 90℃heating, and their polymeric anthocyanin was unstable and easy to be degraded by heating. As to antioxidiant activities, scavenging activity of DPPH
radical increased by increasing heating time and show the same tendency with fraction Ⅱ, which was collected through purified by gel chromatography. Contrastly, FRAP decreased by increasing the heating time. There was no significant difference between pH1 and pH2 roselle anthocyanin model system in all the DPPH, FRAP and TEAC assay. Among the samples of mulberry wine, the wine aged for one year showed the highest copigmentation and polymerization. Scavenging activity of DPPH radical was increased by increasing aging time, while FRAP showed opposite tendency. FractionⅡ purified from gel chromatography showed highest activity of TEAC . According to the results showed above, we may come to the conclusion that in Roselle and mulberry wine, scavenging activity of DPPH radical and FRAP were actually related to degree of polymerization of their anthocyanin.

中文摘要…………………………………………………………...I
英文摘要………………………………………………………….III
誌謝……………………………………………………………….Ⅴ
目錄……………………………………………………………..VⅡ
圖索引…………………………………………………………..XⅢ
表索引…………………………………………………………..XⅥ
壹、前言……………………………………………………………1
貳、文獻回顧……………………………………………………...4
一、花青素之介紹…………………………………………...4
(一)花青素的結構……………………..………………..5
(二)影響花青素穩定之因子..…………………..………7
1.花青素本身結構…………………………………7
2.溫度………………………………………………7
3.光…………………………………………………7
4.氧氣……………………………..………………..8
5.醣類…………….……………..………………….8
6.pH值…………..…………………………………8
7.醯基化………….………..……………………….9
二、花青素的共呈色與聚合………………………………..10
(一)共呈色(Copigmentation)的發現…..……..…..……10
(二)共呈色的定義和特性……………………………..10
(三)共呈色作用的種類和發生機制……………..……11
(四)影響花青素之共呈色作用的因子……..…………13
1.花青素的種類……………………….…….……13
2.花青素的濃度…………………………..………13
3.共呈色分子的種類………………….….………15
4.共呈色分子的濃度……………………….…….15
5.pH值……………………………………………15
6.金屬離子的影響……………………………..…16
(五)聚合作用(Polymerization) ……………………..…17
三、自由基的定義和種類及與人類疾病的關係………….20
(一)自由基的定義和種類……………………………..20
1.超氧自由基(Superoxide radical)：O2-‧….……...20
2.過氧化氫(Hydrogen peroxide)：H2O2..................21
3.羥基自由基(Hydroxyl radical)：‧OH…………..21
4.單重態氧(Singlet oxygen)：1O2……………...…22
5.脂質過氧化物(Lipid hydroperoxide)：ROC‧…..22
(二)自由基與人類疾病的關係………..……...……….22
四、抗氧化劑的定義及其作用機制………….……………24
(一)抗氧化劑的定義………………………………..…24
(二)抗氧化劑的作用機制………………………..……24
1.抑制型(Inhibitor) …………………………...….24
2.清除型(Scavenging agent)……………….……..25
3.還原型(Reducing agent)………………………..25
4.螯合型(Chelator)……………...………………..26
五、酚類與抗氧化活性……………………………………...27
參、材料與方法………………………………………………….30
一、試驗材料………………………………………………...30
(一)原料…………………………………………………30
(二)化學試藥………………….………………………...30
(三)儀器設備……………………………………………31
二、試驗設計………………..…………………………….…32
三、試驗方法……………….………………………………..37
(一)乾燥洛神葵花青素聚合物百分比與抗氧化能力關係之探
討……………..….………………………37
1.乾燥洛神葵樣品製備……………………………37
2.顏色品質測定……………………………………37
3.Copigment和Monomeric及Polymeric花青素之測
定………………………………38
4. DPPH自由基清除能力測定……………………38
(二)洛神葵花青素模擬系統於不同加熱時間下花青素
聚合物百分比與抗氧化能力關係之探討………...39
1.洛神葵花青素模擬系統之製備…………………39
2.顏色品質測定………..…………………..………39
3. Copigment和Monomeric及Polymeric花青素之
測定……………………………………………...40
4.抗氧化活性測定…………………………………40
(1)DPPH自由基清除能力測定………………...40
(2)還原力之測定…………………………….….40
(3)Trolox當量之抗氧化能力測定……………...40
5.不同花青素聚合物之分離純化..………………..41
(1)液相層析法…………………………………..41
(2)高效能液相層析儀分析酚類化合物………..42
(三)桑椹酒於不同貯存時間下花青素聚合物百分比
與抗氧化能力關係之探討……………..…..….…..43
1.桑椹花青素模擬系統之製備…………………..43
2.桑椹酒一般性質分析…………..……………....43
3.桑椹酒顏色品質測定……………..………..…..44
4.花青素之Copigment和Monomer及Polymer
之測定………………………………………….45
5.抗氧化活性測定……………..…………………45
6.不同花青素聚合物之分離純化………………..45
(四)統計分析…………………………………………..45
肆、結果與討論………………………………………………….46
一、乾燥洛神葵於貯存期間花青素聚合物百分比與抗氧化能力之變
化…………………………………………….46
(一)乾燥及貯存溫度對乾燥洛神葵在貯存期間花青素聚合物百分比之影
響…………………………….46
(二)乾燥洛神葵花青素聚合物百分比與抗氧化活性之相關
性…………………………………………….48
二、洛神葵花青素模擬系統於加熱過程中其聚合物百分比與抗氧化能力之
變化………………………………….53
(一)加熱時間對不同pH值之洛神葵花青素模擬系統顏色品質之影
響…………………………………….53
(二)加熱時間對不同pH值之洛神葵花青素聚合物百分比之影
響………………………………………….59
(三)加熱時間對洛神葵花青素抗氧化能力之影響..…61
(四)洛神葵花青素經加熱及膠體過濾後各區分顏色品質之變
化………………………………………….66
(五)洛神葵花青素模擬系統經加熱及膠體過濾區分後各區分物抗氧化能力
之變化…………………….72
(六)乙醛對洛神葵花青素共呈色的影響……………..75
三、不同貯存時間之桑椹酒其聚合物百分比與抗氧化能力之變
化………………………………………………….82
(一)貯存時間對桑椹酒品質之影響…………………..82
(二)貯存時間對桑椹酒聚合物百分比之影響………..87
(三)貯存時間對桑椹酒抗氧化能力之影響…………..89
(四)不同貯存時間之桑椹酒經膠體過濾純化後各區分顏色品質之變
化…………………………………...93
(五)不同貯存時間之桑椹酒經膠體過濾處理後各區分之抗氧化能力之變
化………………………….…100
(六)桑椹酒經膠體過濾後各區分之酚類化合物……105
伍、結論………………………………………………………….109
陸、參考文獻…………………………………………………….111

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