臺灣博碩士論文加值系統

黑豆（Black Soybean）屬於豆科（Fabacae或Leguminosae），
蝶形花亞科（Papilionaceae），大豆屬（Glycine max L. Merril），黑皮種，依其子葉顏色可分為青仁黑豆與黃仁黑豆兩種。本研究以青仁黑豆為原料，以六種加熱處理，包括130℃烘烤 (3或5分鐘)、100 ℃培炒 (8或10分鐘) 及微波中火加熱 (1或2分鐘)，分析黑豆水萃取物抗氧化物質濃度（包括總類黃酮含量、總酚類含量及總花青素含量）和抗氧化活性 (包括清除DPPH自由基能力、亞鐵離子螯合能力及還原力)。結果顯示，以微波2分鐘和烘烤5分鐘處理組，黑豆水萃取物總酚類含量較高（分別為5.90±0.22和5.68±0.08 mg gallic acid / g）；總類黃酮則以烘烤5分鐘處理組最高（39.67±0.27 mg quercetin / g）；微波2分鐘及烘烤5分鐘處理組則含較多總花青素（分別為5.66±0.55和5.23±0.08 mg / g）。以烘烤、培炒和微波三種不同的加熱處理黑豆後發現，130℃烘烤組之黑豆水萃取物具最高含量之抗氧化物質，故進一步以130℃烘烤3分鐘和5分鐘之黑豆分別進行抗氧化活性分析。結果顯示，黑豆經130℃烘烤5分鐘後其抗氧化活性較130℃烘烤3分鐘者佳。黑豆酵素分析方面，130℃烘烤5分鐘之黑豆比未加熱的黑豆有較低的過氧化酶活性卻有較高的多酚氧化酶活性。綜合以上結果，採用130℃烘烤3分鐘和5分鐘之黑豆以100℃的熱水浸泡20分鐘製備黑豆茶湯並進一步評估茶湯品質，分析項目包括：混濁度、色澤及官能品評。與未預熱處理的黑豆茶湯相比，烘烤過程使得黑豆茶湯之 L*、 a* 及 b* 值提升。烘烤5分鐘之黑豆茶湯的 L*、 a* 及 b* 值比未預熱處理之黑豆泡製的茶湯較高，但較市售預熱處理之黑豆泡製的茶湯低。並且130℃烘烤5分鐘泡製的茶湯較烘烤3分鐘之茶湯顏色顯得深濃。130℃烘烤5分鐘黑豆之茶湯於所有感官品評項目 (顏色、口感、香氣、風味及整體接受度) 分數最高，顯示本研究所開發之黑豆茶湯受品評人員接受度高。鑒於130℃烘烤5分鐘黑豆之茶湯高抗氧化活性，本研究以動物模式探討黑豆茶湯對化學性肝損傷之保護能力。
Sprague-Dawely (SD) 大鼠以隨機方式分為控制組、高濃度黑豆茶湯（1000 mg/kg）組、四氯化碳（Carbon tetrachloride, CCl4）組、CCl4＋silymarin（200 mg/kg）組、CCl4＋低濃度黑豆茶湯（100 mg/kg）組及CCl4＋高濃度黑豆茶湯（1000 mg/kg）組，實驗為期九週。與控制組比較，CCl4誘發肝損傷組血清中Glutamate oxaloacetate transaminase（GOT）及Glutamate pyruvate transaminase（GPT）值明顯升高（p<0.05），而肝臟中抗氧化物質還原型麩胱甘肽（GSH）和氧化型麩胱甘肽（GSSG）濃度和抗氧化酵素Superoxide dismutase（SOD）、Glutathione peroxidase（GPx）、Glutathione reductase（GR）及Catalase（CAT）活性則顯著降低 (p < 0.05)，而給予肝損傷治療用藥silymarin、低和高濃度黑豆茶湯之SD大鼠其血清中GOT及GPT值較CCl4組顯著下降，且肝臟中GSH和GSSG濃度、SOD、GPx、GR及CAT活性顯著高於CCl4組。另未以CCl4誘發肝損傷之大鼠給予高濃度黑豆茶湯後，可顯著提升其肝臟中抗氧化物質及SOD、GPx和GR活性。
肝臟病理變化結果顯示CCl4組肝臟中心帶周圍之肝細胞脂質沉積及結締組織增生，且形成嚴重脂肪肝進而導致肝纖維化之發生；而給予silymarin治療或補充黑豆茶湯可降低脂質堆積且無肝纖維化之發生。動物實驗結果顯示，130℃烘烤之黑豆藉其高抗氧化活性，降低四氯化碳造成之肝損傷進而達到護肝效果。因此，本研究製備之黑豆茶湯，確實值得開發成護肝保健產品。

Black soybean (Glycine max L. Merril) has black seedcoat and its cotyledon color is cyan or yellow. In this study, cyan cotyledon black soybean was used as raw material. Six different pre-heating methods including baked at 130℃for 3 or 5 minutes, stir-fried at 100℃for 8 or 10 minutes and microwave heated at medium power for 1 or 2 minutes were applied to evaluate the effects of the pre-heating methods on the antioxidants concentrations (including the total flavonoid content, total phenolic content and total anthocyanins content) and antioxidant activities (including DPPH radical scavenging activity, ferrous ion chelating power and reducing power) in the water extracts from black soybean. The results indicated that microwave heated for 2 minutes and baked at 130℃ for 5 minutes groups had high total phenolic content (5.90 ± 0.22 and 5.68 ± 0.08 mg gallic acid / g, respectively). The baked at 130℃ for 5 minutes group had high total flavonoid content (39.67 ± 0.27 mg quercetin / g). Moreover, the microwave heated for 2 minutes and baked at 130℃ for 5 minutes groups had high total anthocyanins content (5.66 ± 0.55 and 5.23 ± 0.08 mg / g, respectively). In the six different pre-heating methods, it was found that black soybean baked at 130℃ have the highest antioxidants levels, for this reason, the subsequent analysis of antioxidant activity was focus on the water extract from black soybean baked at 130℃ for 3 and 5 minutes. The results showed that water extract from black soybean baked at 130℃for 5 minutes had better antioxidant activity than water extract from black soybean baked at130℃ for 3 minutes. Black soybean baked at 130℃ for 5 minutes had lower peroxidase but higher polyphenol oxidase activities than unheated black soybean. Black soybean tea was prepared from black soybean baked at 130℃ for 5 minutes and 3 minutes by the immerge of hot water at 100℃ for 20 minutes, and its color and sensory properties were evaluated. Compared to unheated black soybean, the data showed that baking process increased L*, a* and b* values of black soybean tea. L*, a* and b* values of black soybean tea baked at 130℃ were higher than black soybean tea made from unheated black soybean but lower than black soybean tea made from commercial pre-heated black soybean. Black soybean tea baked at 130℃ for 5 minutes have darker colors than baked at 130℃ for 3 minutes. Black soybean tea made from black soybean baked at 130℃ for 5 minutes had the highest scores in all sensory properties (color, Mouth - feel, aroma, flavor and overall acceptability). Since the water extract from black soybean baked at 130℃ for 5 minutes had high antioxidant activity, the hepatoprotective effect of black soybean tea baked at 130℃ for 5 minutes against carbon tetrachloride (CCl4)-induced hepatotoxicity was evaluated using animal model.
Sprague-Dawely rats were randomly divided into control group, high concentrations of black soybean tea (1000 mg / kg) group, carbon tetrachloride (CCl4) group, CCl4 + silymarin (200 mg / kg) group, CCl4 + low concentrations of black soybean tea (100 mg / kg) group and CCl4 + high concentration of black soybean tea (1000 mg / kg) group, the experimental period of nine weeks. Compared to control group, CCl4 group increased the activities of serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), but decreased the glutathione (GSH) and oxidized glutathione (GSSG) concentration, as well as the antioxidant enzymes activities including superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and catalase (CAT) in the liver. Treatment with silymarin, or low and high concentrations of black soybean tea could significantly decrease the serum GOT and GPT levels but enhance the GSH and GSSG levels, as well as increase the activities of SOD, GPx, GR and CAT in the liver of CCl4–treated rats. Treatment with high concentrations of black soybean tea alone without CCl4 significantly improved antioxidants levels and SOD, GPx and GR activities in the liver.
Changes of results the pathologic indicated that lipid accumulation and connective tissues proliferation surround the hepatocytes of central zone was found in CCl4-treated rats. The hepatic toxicity caused by CCl4 resulted in serious fatty liver and liver fibrosis in CCl4-treated rats. However, the supplement of silymarin or black soybean tea could reduce the accumulation of lipid in the liver and no occurrence of hepatic fibrosis. Animal experiments confirmed that black soybean tea baked at 130℃ for 5 minutes declined carbon tetrachloride induced liver injury due to the ability to enhance antioxidant levels and antioxidant enzymes activities in the tested animals. Thus, black soybean tea prepared in this study is worth of developing into health drink products to improve liver function.

中文摘要……………………………………………………………………………………………… I
英文摘要…………………………………………………………………………………………… IV
序言………………………………………………………………………………………………… VIII
目次……………………………………………………………………………………………………… X
表次…………………………………………………………………………………………………… XII
圖次………………………………………………………………………………………………… XIII
壹、文獻回顧……………………………………………………………………………………… 1
一、黑豆 ………………………………………………………………………………………… 1
二、自由基與活性氧 ……………………………………………………………………… 3
三、肝臟 ………………………………………………………………………………………… 11
四、水飛薊 …………………………………………………………………………………… 22
貳、材料與方法………………………………………………………………………………… 25
參、結果與討論………………………………………………………………………………… 49
一、預熱處理對黑豆水萃取物抗氧化能力之影響………………………… 49
二、預熱處理黑豆多酚氧化酶及過氧化酶活性…………………………… 51
三、預熱處理黑豆茶湯品質評估…………………………………………………… 52
四、預熱處理黑豆茶湯之護肝作用……………………………………………… 55
肆、結論………………………………………………………………………………………62
伍、表………………………………………………………………………………………………… 63
陸、圖………………………………………………………………………………………………… 70
柒、參考文獻……………………………………………………………………………………… 89

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