臺灣博碩士論文加值系統

蝶豆花(Butterfly pea)，學名為Clitoria ternatea，花瓣中富含醯基花青素(飛燕草素)、酚酸及類黃酮等，具有高抗氧化活性，是一種非常有潛力的機能性作物。根據文獻，糖化酵素(α-glucosidase及α-amylase)是人體參與碳水化合物(雙糖、多糖及澱粉)代謝時的作用酵素，藉由糖化酵素的抑制，能有助於減少葡萄糖之吸收，來降低餐後血糖，達到降血糖之功效，其中，多酚類化合物被認為是具有抗醣化能力的天然來源。然而，本土蝶豆花在抗糖化能力方面的相關研究甚少，故本研究利用屏東地區栽種之蝶豆花為原料，進行糖化酵素抑制能力之探討，以不同濃度乙醇萃取，並分別進行抗糖化能力(α-glucosidase及α-amylase)、抗氧化成分(總酚、總類黃酮及總花青素)、抗氧化能力(DPPH、FRAP)等分析。最後則利用蝶豆花進行發酵，來探討抗糖化能力及抗氧化力，並分析在發酵過程中SOD-like活性及多酚類化合物含量之變化。
結果顯示，50% EtOH蝶豆花萃取液具有最高的α-glucosidase抑制率(57.65%)，蝶豆花水萃液則具有最高的α-amylase抑制率(70.89%)，而在抗氧化成分及能力方面，總酚、總類黃酮及總花青素最高含量分別為3097、813及1035 mg/100g d.w.，並且皆有不錯的DPPH自由基清除能力及FARP三價鐵還原能力。經由HPLC分析多酚類化合物後發現，蝶豆花中具有抗糖化能力的主要酚類化合物包含Gallic acid、Chlorogenic acid及Ferulic acid，而類黃酮化合物則為Morin、Rutin、Vitexin及Quercetin，對於抗糖化能力具有相當大的潛力。
而發酵過後，皆能提升糖化酵素抑制能力，其中，α-glucosidase與α-amylase的抑制率最高分別達到67%與90%。而抗氧化活性如SOD-like、DPPH及FRAP分別上升1.5、1.13及1.11倍，此外，HPLC分析顯示，大部分多酚類化合物皆有提升，各約提高1.18至4.39倍不等。且經由統計分析結果發現，蝶豆花抗糖化能力與總酚、總類黃酮及SOD-like活性具有高度相關，其中，具有高度相關的多酚類化合物包含Kaempferol、Quercetin、Epicatechin、Rutin及Morin等。
而在最後利用田口法所得的最適蝶豆花酵素飲料配方之儲藏安定性試驗，其分析項目包含SOD-like、總酚含量及總類黃酮含量，由實驗結果可以得知，個別分析項目殘留80%時，在5℃中各可以存放107.40、135.40及103.82週，皆具有相當不錯的儲藏安定性。
綜合上述結論，蝶豆花是一種具有抗醣化潛力之天然食材，其中富含許多機能性成分及高抗氧化力，且在經過發酵的加工方式後，能夠釋放出更多多酚類化合物來提高其蝶豆花抗糖化之能力以及抗氧化力，在儲藏安定性試驗中也具有良好的保存能力。對於未來蝶豆花天然來源抗糖化能力之產品開發，可以得到完善的利用，並增加其在保健領域方面之利用性，來提高蝶豆花經濟價值。

Butterfly pea is one kind of functional plants. Its petals are rich in acylated anthocyanin, which derived from delphinidin, and in phenolic acids and flavonoids as antioxidant. From the previous studies, metabolism of carbohydrates(starch oligosaccharides and disaccharides) in human body was carried on by α-glucosidase and α-amylase. If glucose absorptions and postprandial blood glucose was inhibited can be decreased the α-glucosidase and α-amylase activity. In addition, polyphenolic compounds are considered to be the effective natural source of antiglycative ability. However, very few studies about antiglycative ability of local butterfly pea effect can be found. So in this study, different solvent were used to extract local butterfly pea and analyze their antiglycative ability (α-amylase and α-glucosidase). Antioxidant composition (total phenolic, total flavonoid and total anthocyanin) and antioxidant capacity(DPPH, FRAP) was also be compared. Finally, fermentation was used to increase the antiglycative ability and the change of antioxidant activity, SOD-like activity and polyphenols content during the fermentation were investigated.
The result showed that 50% EtOH extract is the best for α-glucosidase inhibition(57.65%) and water extract is the best for α-amylase inhibition (70.89%). The content of total phenol, total flavonoid and total anthocyanin may reach 3097, 813, and 1035 mg/100g respectively. We also found that gallic acid、chlorogenic acid and ferulic acid were the major phenolic acid ; morin、rutin、vitexin and quercetin were the major flavonoids in butterfly pea by HPLC analysis.
After fermentation, the antiglycative ability can effectively increase to 67% and 90% respectively. In the part of antioxidant activity, significanly increase in SOD-like activity, DPPH sceavenging and FRAP was found. According to HPLC analysis, individual polyphenol increased around 1.18 to 4.39 times when compared with the sample without fermentation. Statistical analysis showed that the antiglycative ability was highly correlated with total phenol followed by the total flavonoids. The kaempferol、quercetin、epicatechin、rutin and morin were highly correlated with antiglycative ability especially.
The last stage used Taguchi method to produce a product, butterfly pea enzyme beverage, And use this product to storage stability test. Analysis item include SOD-like、total phenolic and total flavonoid. Result show that butterfly pea enzyme beverage at 5℃ can be stored for 107.40、135.40 and 103.82 weeks respectively.
Base on the above results, the butterfly pea is a natural food with antiglycative ability, which is rich in functional compound and high antioxidant. After fermentation processing, it can release more polyphenolic compounds to enhance antiglycative ability and antioxidant capacity. Butterfly pea also showed great storage stability. For the future, development of the natural source to antiglycative ability from butterfly pea, can be fully utilized and development in the field of health and enhanced economic value.

中文摘要 I
Abstract III
誌謝 V
目錄 VI
圖索引 X
表索引 XII
一、前言 1
二、文獻回顧 2
2.1 蝶豆之介紹 2
2.1.1蝶豆機能性成分 2
2.1.2蝶豆藥理活性 5
2.2 多酚類化合物 5
2.2.1酚酸(Phenolic acids) 6
2.2.1.1羥基苯甲酸(Hydroxybenzoic acids) 6
2.2.1.2羥基肉桂酸 (hydroxycinnamic acids) 8
2.2.2類黃酮(flavonoids) 10
2.2.2.1黃酮(Flavones) 12
2.2.2.2黃酮醇(Flavanonols) 12
2.2.2.3黃烷酮(Flavanones) 12
2.2.2.4黃烷醇(Flavanols) 13
2.2.2.5異黃酮(Isoflavones) 13
2.2.3花青素(Anthocyanins) 13
2.2.3.1 花青素結構及生理活性 14
2.2.3.2醯基花青素(Acylated anthocyanin) 17
2.3糖化酵素 18
2.3.1 α-葡萄糖苷酶(α-Glucosidase) 18
2.3.2 α-澱粉酶(α-Amylase) 18
2.3.3抗糖化能力與調控血糖 19
2.3.4天然物抑制糖化酵素 20
2.3.4.1各類多酚類化合物抗糖化能力之IC50 22
2.3.4.2酚酸抗糖化能力之機制 24
2.3.4.3類黃酮羥基抗糖化能力之機制 27
2.6發酵之應用 32
2.6.1發酵對於抗氧化之影響 32
2.6.2超氧歧化酶(SOD) 33
2.6.3超氧歧化酶類似物(SOD-like)應用 35
三、材料與方法 36
3.1試驗材料 36
3.2酵素製備 36
3.3試驗藥品 36
3.4試驗儀器 37
3.5試驗設計 39
3.5.1不同部位及溶劑對於蝶豆抗氧化及抑制糖化酵素活性之影響 39
3.5.2不同比例之蝶豆花在發酵過程對於抗糖化能力及抗氧化力之變化 39
3.5.3蝶豆花酵素最適配方之試驗 40
3.6試驗方法 44
3.6.2糖化酵素抑制率分析 44
3.6.2.1 α-glucosidase 抑制率測定 44
3.6.2.2 α-amylase抑制率測定 44
3.6.3抗氧化成分分析 45
3.6.3.1總多酚含量測定 45
3.6.3.2總類黃酮含量測定 45
3.6.3.4總花青素含量測定 46
3.6.3.3多酚類化合物定性及定量分析 47
3.6.4抗氧化能力分析 49
3.6.4.1 DPPH自由基清除能力測定 49
3.6.4.2 三價鐵還原能力(Frric iron reducing power)測定 49
3.6.4.3清除超氧陰離子能力(SOD-like)測定 50
3.6.5品質分析 50
3.6.5.1色澤分析(Hunter L a b) 50
3.6.5.2可溶性固形物(Total soluble solids, °Brix) 51
3.6.5.3 pH值 51
3.6.5.4酒精度 51
3.6.6田口法(Taguchi method) 51
3.6.7官能品評試驗 51
3.6.8統計分析 54
四、結果與討論 55
4.1. 蝶豆不同部位及溶劑對於糖化酵素活性之分析 55
4.1.1 蝶豆不同部位α-Glucosidase及α-Amylase抑制率 55
4.1.2不同萃取溶劑對於蝶豆花抑制α-Glucosidase及α-Amylase之分析 55
4.1.2.1 α-Glucosidase抑制率 57
4.1.2.2 α-Amylase抑制率 57
4.2蝶豆花不同溶劑萃取液抗氧化成分之分析 60
4.2.1總酚含量(Total phenolic content) 60
4.2.2總類黃酮含量(Total flavonoid content) 63
4.2.3多酚類化合物之定性及定量分析 64
4.2.4總花青素含量(Total anthocyanin content) 70
4.3蝶豆花不同溶劑萃取液抗氧化能力之分析 73
4.3.1 DPPH自由基清除能力 73
4.3.2 FRAP三價鐵還原能力 75
4.4發酵過程中不同比例蝶豆花對於抗糖化能力之影響 77
4.4.1 α-Glucosidase抑制率 77
4.4.2 α-Amylase抑制率 79
4.5發酵過程中不同比例蝶豆花對於抗氧化力之影響 81
4.5.1 SOD-like活性分析 81
4.5.2 DPPH自由基清除能力 84
4.5.3 FRAP三價鐵還原能力 86
4.6發酵過程中不同比例蝶豆花對於抗氧成分之影響 88
4.6.1總酚含量 88
4.6.2總類黃酮含量 91
4.6.3總花青素含量 93
4.6.3多酚化合物之定性及定量 95
4.7相關性分析 99
4.7.1發酵過程糖化酵素抑制活性與抗氧化力之相關性 99
4.7.2發酵過程糖化酵素抑制活性與多酚類化合物之相關性 99
4.8蝶豆花發酵過程中品質分析 102
4.8.1色澤分析 102
4.8.2 pH值 103
4.8.3酒精度 103
4.8.4可溶性固形物°Brix 106
4.9蝶豆花酵素飲料製作 106
4.9.1田口法分析 106
五、結論 111
參考文獻 113
附錄一 132
作者簡介 133

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