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研究生:王志玄
研究生(外文):Jhih-Syang Wang
論文名稱:黑豆的抗氧化能力研究
論文名稱(外文):The antioxidant capacity of black soybean
指導教授:梁致遠梁致遠引用關係吳慧中吳慧中引用關係
指導教授(外文):Ji-Yuan LiangHui-Chung,Wu
學位類別:碩士
校院名稱:銘傳大學
系所名稱:生物科技學系碩士班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:83
中文關鍵詞:超氧化物歧化酶自由基還原力黑豆福林試劑
外文關鍵詞:black soybeanreducing powerFolin-Ciocalteu methodSuperoxide Dismutase (SOD)free radicals
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以黑豆為研究對象,探討黑豆的種仁及種皮的抗氧化能力,以作為保健食品開發的基礎。Folin-Ciocalteu方法常作為總多酚的測試方法,以沒食子酸作為總多酚標準品,沒食子酸先加強鹼的方法,會產生新的化合物,影響Folin-Ciocalteu方法的測試結果。利用不同的pH值以及不同濃度的乙醇做萃取,藉由總多酚的測定,找出最適萃取條件。以pH3 、70%乙醇萃取黑豆之總多酚為57.22±3.94 mg GAE/mL效果最好,因此作為黑豆萃取的最佳條件做後續抗氧化能力測試,無論是多酚類含量、還原力、清除DPPH自由基能力及SOD活性,黑豆種皮都較黑豆仁、黃豆皮、黃豆仁為高。以E.coli(pGEM-7zf(-))抽出的質體DNA,添加核黃素並照藍光,產生的超氧自由基會破壞DNA超螺旋結構,添加10,000μg/mL濃度的黑豆皮萃取液,則能保護質體DNA的超螺旋結構免於破壞。經發芽的黑豆,黑豆種皮的多酚類含量、還原力、清除DPPH自由基能力及SOD活性,隨著發芽時間愈長其抗氧化能力逐漸降低。
The goal of this study was to explore the antioxidative capacity of both seed kernel and seed coat of black beans, forming the basis of future healthy food development. Folin-Ciocalteuz method has been routinely used to determine the cuntent of total polyphenols, with gallic acid being need as the standard, However, upon reaction with strong base, Gallic acid can produce new compounds and this way affect the testing results of Folin-Ciocalteu method. Using different pH values and extraction at different concentrations by ethanol, the optimum extraction condition was established as shown by the highest content of total polyphenols determined by extraction 70% ethanol at pH 3.0,total polyphenols of black beans of 57.22±3.94 mg GAE/mL were shown to be the highest content. Therefore this condition can be used for the determination of black bean antioxidative capacity, due to optimal conditions of extraction and applied in follow-up tests. In terms of polyphenols contents, reducing power, DPPH radical scavenging and SOD activity, seed coat of black bean was shown to be higher than seed kernel of black beans, seed coat and seed kernel of soybean beans. To extract plasmid DNA(pGEM-7zf (-)),we added riboflavin and shined blue-ray, this generated superoxide free radical and damaged the structure of plasmid DNA supercoiling. We added black bean seed coat extracts concentrations of 10,000 μ g/mL to protect the integrity of plasmid DNA. Polyphenols contents, reducing power, DPPH radical scavenging ability and SOD activity of seed coatwere found to decrease along with the antioxidative capacity throughtout the sprouting of black beans.
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摘要
Abstract
壹、 第一節 鹼催化對Folin-Ciocalteu試劑檢測總多酚含量的影響
一、 前言
二、 材料與方法
1. 藥品
2. 儀器
3. 實驗方法
三、 實驗結果
1. 鹼對沒食子酸吸收光譜的影響
2. 添加試劑的順序對沒食子酸製作檢量線的影響
3. 以兒茶素加酸的方式,產生鍵反應,驗證Folin-Ciocalten 試劑與碳酸鈉的順序對沒食子酸量測的影響
四、 討論
五、 參考文獻
貳、 第二節 黑豆種皮及種仁抗氧化能力的研究
一、 前言
二、 材料與方法
1. 黑豆來源
2. 藥品
3. 儀器
4. 菌株
5. 實驗方法
三、 實驗結果
1. 乙醇最適合的萃取條件
2. 黑豆皮的總多酚、 清除DPPH自由基能力、還原力以及SOD活性測試皆優於黑豆仁、黃豆仁及黃豆皮。
3. 黑豆皮對plasmid DNA具保護作用
四、 討論
五、 參考文獻
參、 第三節 發芽對黑豆抗氧化能力的影響
一、 前言
二、 材料與方法
1. 黑豆
2. 藥品
3. 儀器
4. 實驗方法
三、 實驗結果
四、 討論
五、 參考文獻
肆、 第四節結論
附錄
壹、第一節 鹼催化對Folin-Ciocalteu試劑檢測總多酚含量的影響
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2.張清安、范學輝、張志琪、馬學燕、陳瑜、王軍:沙苑子提取物中多酚含量的測定。食品科學2010,31:178-181。
3.Alonso Borbalán ÁM, Zorro L, Guillén DA, Barroso CG: Study of the polyphenol content of red and white grape varieties by liquid chromatography–mass spectrometry and its relationship to antioxidant power, J Chromatogr A 2003, 1012: 31–38.
4.Ao C, Li A, Elzaawely AA, Xuan TD, Tawata S: Evaluation of antioxidant and antibacterial activities of Ficus microcarpa L. fil. Extract, Food Control 2008, 19: 940-948.
5.Chen HY , Yen GC: Antioxidant activity and free radical-scavenging capacity of extracts from guava (Psidium guajava L.) leaves, Food Chem 2007, 101: 686-694
6.Chen HY, Lin YH, Hsieh CL: Evaluation of antioxidant activity of aqueous extract of some selected nutraceutical herbs, Food Chem 2007, 104: 1418–1424.
7.Číž M, Čížová H, Denev P, Kratchanova M, Slavov A, Lojek A: Different methods for control and comparison of the antioxidant properties of vegetables, Food Control 2010, 21: 518–523
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11.Faller ALK, Fialho E: The antioxidant capacity and polyphenol content of organic and conventional retail vegetables after domestic cooking, Food Research International 2009,42: 210–215.
12.Friedman M, Jurgens HS: Effect of pH on the stability of plant phenolic compounds, J Agric Food Chem 2000: 48: 2101-2110.
13.Folin O, Denis W: A Colormetric method for the determination of phenols ( and phenol derivates) in urine. J Bio Chem 1915, ⅩⅩⅡ, No 2.
14.Gahler S, Otto K, Bohm V : Alterations of Vitamin C, toal phenolics, and antioxidant capacity as affected by processing tomatoes to different products, J Agric Food Chem 2003, 51:7962-7698.
15.Huang D, Ou B, Prior R: The Chemistry behind antioxidant capacity assays, J. Agric Food Chem 2005, 53: 1841-1856
16.Hussein AMS, Shedeed NA, Abdel-Kalek HH, Shams El-Dinb MHA: Antioxidative, Antibacterial and Antifungal Activities of Tea Infusions from Berry Leaves, Carob and Doum, Pol J Food Nutr Sci 2011, 61: 201-209.
17.Katsube T, Tabata H, Ohta Y,Yamasaki Y, Anuurade E, Shiwakuk K, Yamane Y: Screening for antioxidant activity in edible plant products: comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin-Ciocalteu assay, J Agric Food Chem 2004, 52: 2391–2396.
18.Lee MH, Jiang CB, Juan SH, Lin RD, Hou WC: Antioxidant and heme oxygenase-1 (HO-1)-induced effects of selected Taiwanese plants, Fitoterapia 2006, 77: 109–115
19.Maksimović Z, Dorđe M, Kovačević: Polyphenol contents and antioxidant activity of Maydis stigma extracts, Bioresour Technol 2005, 96: 873–877.
20.Pinelo M, Rubilar M, Sineiro J, Núñez MJ: Extraction of antioxidant phenolics from almond hulls (Prunus amygdalus) and pine sawdust (Pinus pinaster), Food Chem 2004, 85: 267–273.
21.Sachan NK, Arif M, Zaman K, Kumar Y: Anti-inflammatory, analgestic and antioxidant potential of the stem bark of spondlas mangifera willd. Arch Biol Sci Belgrade 2011, 63: 413-419.
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貳、第二節 黑豆種皮及種仁抗氧化能力的研究
1.徐金瑞、張名位、劉興華、張瑞芬、孫玲:黑大豆種皮花色苷體外抗氧化活性研究。營養學報,2007,29:54-57
2.連大進:台灣黑豆的利用與生產展望。農業世界,1995,147:9-42
3.叢建民:黑豆的營養成分分析研究。食品工業科技,2008,4:262-268
4.Beauchamp C, Fridovich I: Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 1971, 44: 276-287.
5.Choung MG, Baek IY, Kang ST, Hang WY, Shin DC, Moon HP, Kang KH: Isolation and Determination of Anthocyanins in Seed Coats of Black Soybean (Glycine max (L.) Merr.). J. Agric. Food Chem. 2001, 49: 5848-5851.
6.Duh PD, Yen GC, Yen WJ, Chang LW: Antioxidant effects of water extracts fromb Barley (Hordeum vulgare L.) prepared under different roasting temperatures. J Agric Food Chem 2001, 49 , 1455–1463
7.Gahler S, Otto K: Bo¨hm V: Alterations of vitamin C, total phenolics, and antioxidant capacity as affected by processing tomatoes to different products. J. Agric. Food Chem 2003,51, 7962-7968.
8.Hou WC, Lin RD, Cheng KT, Hung YT, Cho CH, Chen CH, Hwang SY, Lee M H: Free radical-scavenging activity of Taiwanese native plants. Phytomedicine 2003, 10, 170–175.
9.Takanori T, Kaoru S, Katsumi O, Shunro K, Toshihiko O: Inhibition of lipidperoxidation and the active oxygen radical scavenging effect of anthocyanin pigments isolated from Phaseolus vulgaris L. Biochemical Pharmacology 1996, 52: 1033-1039.
10.Xu JR, Zhang MW, Liu XH, Liu ZX, Zhang RF, Sun L, Qiu LJ: Correlation between antioxidation and the content of total phenolics and anthocyanin in black soybean accessions. Agricultural Sciences in China 2007, 6: 150-158.
參、第三節 發芽對黑豆抗氧化能力的影響
1.Beauchamp C, Fridovich I: Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 1971, 44: 276-287.
2.Duh PD, Yen GC, Yen WJ, Chang LW: Antioxidant effects of water extracts fromb Barley (Hordeum vulgare L.) prepared under different roasting temperatures. J Agric Food Chem 2001, 49 , 1455–1463
3.Gahler S, Otto K: Bo¨hm V: Alterations of vitamin C, total phenolics, and antioxidant capacity as affected by processing tomatoes to different products. J. Agric. Food Chem 2003,51, 7962-7968.
4.Hou WC, Lin RD, Cheng KT, Hung YT, Cho CH, Chen CH, Hwang SY, Lee M H: Free radical-scavenging activity of Taiwanese native plants. Phytomedicine 2003, 10, 170–175.
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