過去研究顯示，將豆類榨汁後所剩餘的廢棄物豆渣，含有高量膳食纖維、大豆胜肽及植物固醇等機能性物質，豆渣經水解酵素分解後，發現其胜肽具有抗氧化與抑制血管收縮素與抑制發炎的能力。以往的研究表明，豆渣可具有促進健康的功能性成分。因此，本研究的目的是調查豆渣的健康功能性，尤其是在抗氧化，抗發炎和抗疲勞的各方面。
本實驗第一階段主要針對黃豆渣及黑豆渣分解後的功能性進行討論，黃豆渣及黑豆渣進行酵素水解及經超臨界萃取所獲得之水解物進行抗氧化活性測定、血管收縮素轉化酶抑制活性之檢測及 RAW 264.7 巨噬細胞抗發炎功能之評估。第二階段便針對棒球運動員為對象，探討運動訓練期間豆渣的補充對疲勞指標與肌肉損傷指標的影響。
第一階段結果顯示，黃豆渣及黑豆渣進行酵素水解方式所獲得之水解物，皆具有抗氧化及抗發炎能力，經Alcalase分解黃豆渣水解蛋白萃取物能力較佳；而降低高血壓能力部份則以黑豆渣水解蛋白萃取物較佳。另一方面，由不同壓力萃取之高壓水相及低壓油相之黃豆渣與黑豆渣超臨界萃取物，皆具有抗發炎能力，經LPS誘導RAW264.7巨噬細胞產生其發炎物質中，抑制一氧化氮產生情形，以高壓水相豆渣超臨界萃取物抑制能力較佳，抑制IL-1β能力則無顯著差異，PGE2產生情形以油相黑豆渣超臨界萃取物抑制能力較佳。整體而言代表黃豆渣及黑豆渣具有機能性成分對於發炎能力具有抑制效果。
第二階段提供23名男性18-23歲棒球隊員在夏季進行為期8週運動訓練期間，食用黑豆渣及黃豆渣成分的餅乾後，血清總膽固醇，三酸甘油三酯，低密度脂蛋白膽固醇和γ–麩胺酸轉移酶在運動訓練過程中顯著下降。相關疲勞指標血氨及游離脂肪酸、肌酸激酶、肌紅蛋白及天門冬胺酸轉移酶與未運動比較下有顯著差異，初期訓練時，肌酸激酶、肌紅蛋白, 血氨及游離脂肪酸顯著增加，說明在運動訓練時發生疲勞及肌肉損傷，然而，經補充豆渣後，相關疲勞及肌肉損傷的標誌有顯著下降，表示豆渣可顯著減少肌肉損傷，並在運動過程中延緩疲勞，豆渣餅乾具有作為運動營養補充品的潛力。

Soybean has nutritional value and health functions. Okara, the soybean residue from soy milk production, also contains micronutrients and functional components such as isoflavones and phytosterols. Previous researches demonstrated that okara could be useful as a functional ingredient with health-promoting attributes. Therefore, the objective of this study was to investigate the health functionality of okara, especially in aspects of anti-oxidation, anti-inflammation, and anti-fatigue.
In the first stage of this study, auther evaluated and discussed 1) the functionality of soybean okara and black soybean okara; 2) the analysis of products of okara by the hydrolytic enzymes and supercritical fluid extraction; and 3) the ACE inhibitory activity and the anti-inflammation capability of LPS-induced RAW 264.7 macrophages. In the second stage of this study, auther studied the effect of okara supplementation on fatigue and muscle damage in university athletes over an exercise training season.
The results of this study showed that the extracts of both soybean okara and black soybean okara have the capability of anti-oxidation and anti-inflammation. Okara hydrolysates of soybean and black soybean okara by alcalase have a significant functionality, and hydrolysates of black soybean okara have better capability to ACE inhibitory activity. In addition, the extracts of soybean okara and black soybean okara produced by supercritical fluid extraction have better anti-inflammatory capability.
Finally, a total of 23 male university baseball players aged 18-23 years attending a summer intensive exercise training program completed an 8-week exercise training-okara intervention study. The results demonstrated significant differences in blood levels of ammonia, free fatty acids, creatine kinase, myoglobin, and aspartate transferase between baseline and exercise training, indicating that fatigue and muscle damage occurred during exercise training. However, those increased markers of fatigue and muscle damage were significantly decreased after the okara supplementation.

壹、前言 4
1.1研究動機 4
1.2研究目的 5
貳、主要材料文獻整理 6
1.黃豆文獻回顧 6
1-1黃豆簡介 6
1-2黃豆成分分析 6
2.黑豆文獻回顧 8
2-1黑豆簡介 8
2-2黑豆成分分析 9
3.豆渣文獻回顧 9
3-1豆渣簡介 9
3-2豆渣之成分分析 9
3-3豆渣之用途 10
3-4豆渣加工食品 11
3-5豆渣之功能性分析 12
參、主題一：豆渣水解蛋白抗氧化、降血壓及抗發炎功能性 15
1.實驗動機與目的 16
2.實驗材料與方法 18
2-1.實驗架構 18
2-2.實驗材料 19
2-3.試驗藥品 19
2-4.儀器設備 22
2-5.實驗方法 22
3.結果 28
4.討論 40
5.結論 42
6.參考文獻 43
肆、主題二：豆渣超臨界萃取物抗發炎功能性 46
1.實驗動機與目的 47
2.實驗材料與方法 50
2-1.實驗架構 50
2-3.實驗材料 51
2-3.試驗藥品 52
2-5.實驗方法 54
3.結果 59
3-1.豆渣超臨界萃取萃取物 成分分析 59
3-2.豆渣超臨界萃取萃取物 抗發炎功能性 60
4.討論 78
6.結論 80
7.參考文獻 81
伍、主題三：豆渣的保健機能評估：人體試驗探討抗發炎、抗疲勞情形 83
1.實驗動機與目的 84
2.實驗材料與方法 86
2-1.實驗架構 86
2-2.實驗材料 87
2-2-1.受試者與實驗設計 87
2-2-2.餅乾製作 87
2-2-3.運動訓練 88
2-2-4.實驗方法 88
2-2-5.運動員營養問卷 89
2-2-6.血液生化值（實驗樣品及方法） 89
2-2-7.統計分析 90
3.結果 90
3-1.豆渣成分分析及胺基酸分析 90
3-2.豆渣餅乾成分及品評結果 90
3-3.運動員基本資料分析 92
3-4.運動員疲勞因子檢測結果 92
4.討論 101
5.結論 106
6.參考文獻 107

Aluko RE. (2008) Journal of AOAC International. Determination of nutritional and bioactive properties of peptides in enzymatic pea, chickpea, and mung bean protein hydrolysates. 91(4), 947-56.
Atsushi Y, Yoko T, Tetsuo T. (2002) Food Science and Technology Research. Antioxidative Activity of Peptides Prepared from Okara Protein. 8 (4), 357–359.
Burns J, Gardner P. T, O’Neil J, Crawford S, Morecroft I, McPhail D. B, Lister C, Matthews D, MacLean M. R, Lean M. E. J, Duthie G. G, Crozier A. (2000) J. Agric. Food Chem. Relationship among antioxidant activity, vasodilation capacity and phenolic contents of red wine. 48(4), 220–230.
Chen H. M, Muramoto K, Yamauchi F. (1995) Journal of Agricultural and Food Chemistry. Structural analysis of antioxidative peptides from soybean β-conglycinin. 43: 574-578.
Cushman D. W, Cheung H. S, (1971) Biochemical Pharmacology. Spectrometric assay and properties of angiotensin-converting enzyme of rabbit lung. 20: 1637-1648.
Furuta S, Takahashi M, Takahata Y, Nishiba Y, Oki T, Masuda M. (2003) Food Sci. Radical-scavenging activities of soybean cultivars with black seed coats. Technol Res 9:73–75.
Huo R, Zhou O. L, Wang B. X, Tashiro S. I, Onoder S, Ikejima T. (2004) Acta Pharmacologica Sinica. Diosgenin induces apoptosis in HeLa cells via activation of caspase pathway. 25 (8), 1077-1082.
Jahan-mihan A, Smith CE, Hamedani A, Anderson GH.(2011) Nutrition Research. Soy protein-based compared with casein-based diets fed during pregnancy and lactation increase food intake and characteristics of metabolic syndrome less in female than male rat offspring. 31(8), 644-51.
Kim H. J. Tsoy I. Park J. M, Chung J. I, Shin S. C, Chang K. C. (2006) Federation of European Biochemical Societies. Anthocyanins from soybean seed coat inhibit the expression of TNF-R-induced genes associated with ischemia/reperfusion in endothelial cell by NF-κB-dependent pathway and reduce rat myocardial damages incurred by ischemia and reperfusion in vivo. 580:1391–1397.
Kitts DD, Weiler K. (2003) Curr Pharm Des. Bioactive proteins and peptides from food sources. Applications of bioprocesses used in isolation and recovery. 9(16), 1309–23.
Kong X, Guo M, Hua Y, Cao D, Zhang C. (2008 ) Bioresour Technol. Dec . Enzymatic preparation of immunomodulating hydrolysates from soy proteins 99(18), 8873-9.
Kwon S. H, Ahn I. S, Kim S. O, Kong C. S, Chung H. Y, Do M. S, Park K. Y. (2007) J. Med. Food Anti-obesity and hypolipidemic effects of black soybean anthocyanins. 10: 552–556.
Lee C. H, Yang L, Xu J. Z, Yeung S. Y. V, Huang Y, Chen Z. Y. (2005) Food Chemistry. Relative antioxidant activity of soybean isoflavones and their glycosides. 90: 735-741.
Mensor L. L. (2001) Phytotherapy Research. Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. 15: 127-130.
Miller L. N, Rice-Evans C. A, Davis M. J, Gopinathan V, Milner A. (1993) Clinical Sience. A novel method for measuring antioxidant status in premature neonates. 84: 407-412.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans, C. (1999) Free Radical Biology & Medicine. Antioxidant activity applying an improved ABTS radical cation decolorization assay. 26: 1231-1237.
Shih P. H, Yeh C. T, Yen G. C. (2007) J. Agric. Food Chem. Anthocyanins induce the activation of phase II enzymes through the antioxidant response element pathway against oxidative stress-induced apoptosis. 55: 9427-9435.
Tamura Y, Takenaka T. (1999) Nippon Shokuhin Kogyo Gakkaishi . Antioxidative activity of water soluble extracts from okara fermented with Bacillus natto and Rizopus oligosporus. 46: 561-569.
Tsuruki T, Yoshikawa M. (2004) Biosci Biotechnol Biochem. Design of soymetide-4 derivatives to potentiate the anti-alopecia effect. 68: 1139–41.
Wang W, Shen S, Chen Q, Tanga B, He G, Ruan H, Das UN. (2008) Curr Pharm Biotechnol. Hydrolyzates of silkworm pupae (Bombyx mori) protein is a new source of angiotensin I-converting enzyme inhibitory peptides (ACEIP). 9(4),307-14.
Wu J, Ding X. (2001) Journal of Agricultural and Food Chemistry. Hypotensive and physiological effect of angiotensin coverting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats. 49: 501-506.
Xiang Zhen K, Ming Ming G, Yu Fei H, Dong C, Cai Meng Z. (2008) Bioresource Technology. Enzymatic preparation of immunomodulating hydrolysates from soy proteins. 99: 8873–8879.
Xu B, Chang S. (2007) J. Food Sci. A comparative study on phenolic profiles and antioxidant activities of legumes as affected by extraction solvents. 72: S159-S166.
施明智，(2002) ，豆渣中蛋白質之最適萃取條件，台灣農業化學與食品科學40:93-100。
楊開聰，(2002)。以Bacillus natto發酵黑豆製造高血纖維蛋白水解活性及高抗氧化力之納豆製品。九十一年度國科會專題研究計畫成果報告。
Boersma B. J, Barnes S, Kirk M, Wang C. C, Smith M, Kim H, Xu J, Patel R, Darley-Usmar V. M. (2001) Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. Soy isoflavonoids and cancer - metabolism at the target site. 480-481:121-127.
Chan Y. C, Wu C. C, Chan K. C, Lin Y. G, Liao J. W, Wang M. F, Chang Y. H, Jeng K. C. (2009) International Journal of Nanomedicine. Nanonized black soybean enhances immune response in senescence-accelerated mice. 4:27-35.
Choung M. G, Baek I. Y, Kang S. T, Han W. Y, Shin D. C, Moon H. P, Kang K. H. (2001) Journal of Agricultural and Food Chemistry. Isolation and determination of anthocyanins in seed coats of black soybean (Glycine max (L.) Merr.). 49(12), 5848-51.
Gallo D, Ferlini C, Fabrizi M, Prislei S, Scambia G. (2006) Carcinogenesis. Lack of stimulatory activity of a phytoestrogen-containing soy extract on the growth of breast cancer tumors in mice. 27(7), 1404-9.
Gudbrandsen OA, Wergedahl H, Berge RK. (2009) Nutrition. A casein diet added isoflavone-enriched soy protein favorably affects biomarkers of steatohepatitis in obese Zucker rats. 25(5), 574-80.
Huo R, Zhou O. L, Wang B. X, Tashiro S. I, Onoder S, Ikejima T. (2004) Acta Pharmacologica Sinica. Diosgenin induces apoptosis in HeLa cells via activation of caspase pathway. 25 (8), 1077-1082.
Jung K. O, Park S. Y, Park K. Y. (2006) Nutrition. Longer aging time increases the anticancer and antimetastatic properties of doenjang. 22(5):539-45.
Kim J, Kwon C. (2001) Nutrition Research. Estimated dietary isoflavone intake of Korean population based on National Nutrition Survey. 21(7), 947-953.
Moreno J. J, Carbonell T, Sánchez T, Miret S, Mitjavila M. T. (2001) The journal of nutrition. Olive oil decreases both oxidative stress and the production of arachidonic acid metabolites by the prostaglandin G/H synthase pathway in rat macrophages. 131(8), 2145-9.
Quitain A. T, Oro K, Katoh S, Moriyoshi T. (2006) Bioresour Technol. Recovery of oil components of okara by ethanol-modified supercritical carbon dioxide extraction. Sep;97(13):1509-14.
Raso G. M, Meli R, Di Carlo G, Pacilio M, Di Carlo R. (2001) Life Sciences. Inhibition of inducible nitric oxide synthase and cyclooxygenase-2 expression by flavonoids in macrophage J774A.1. 68(8), 921-31.
Shih P. H, Yeh C. T, Yen G. C. (2007) Journal of Agricultural and Food Chemistry. Anthocyanins induce the activation of phase II enzymes through the antioxidant response element pathway against oxidative stress-induced apoptosis. 55(23), 9427-35.
Takahashi R, Ohmori R, Kiyose C, Momiyama Y, Ohsuzu F, Kondo K. (2005) Journal of Agricultural and Food Chemistry. Antioxidant activities of black and yellow soybeans against low density lipoprotein oxidation. 53(11), 4578-82.
Wang B. F, Wang J. S, Lu J. F, Kao T. H, Chen B. H. (2009) Journal of Agricultural and Food Chemistry. Antiproliferation effect and mechanism of prostate cancer cell lines as affected by isoflavones from soybean cake. 57(6), 2221-32.
Xu B. J, Yuan S. H, Chang S. K. (2007) Journal of Food Science. Comparative studies on the antioxidant activities of nine common food legumes against copper-induced human low-density lipoprotein oxidation in vitro. 72(7), S522-7.
Yan S. R, Joseph R. R, Wang J, Stadnyk A. W. (2006) The Journal of Immunology. Differential pattern of inflammatory molecule regulation in intestinal epithelial cells stimulated with IL-1. 177(8), 5604-11.
Yen G. C, Lai H. H. (2003) Journal of Agricultural and Food Chemistry. Inhibition of reactive nitrogen species effects in vitro and in vivo by isoflavones and soy-based food extracts. 51(27), 7892-7900.
Atsushi Y, Yoko T, Tetsuo T. (2002) Food Sci. Antioxidative activity of peptides prepared from okara protein. 8(4), 357–359.
Banister E. W, Allen M. E, Mekjavic I. B, Singh A. K, Legge B, Mutch J. C (1983). European Journal of Applied Physiology. The time course of ammonia and lactate accumulation in blood during bicycle exercise. 51: 195–202.
Banister E. W, Cameron B. J. C (1990) International Journal of Sports Medicine. Exercise-induced hyperammonemia: peripheral and central effects. 11(2), S129-42.
Blomstrand E, Saltin B (2001) American journal of physiology-endocrinology and metabolism. BCAA intake affects protein metabolism in muscle after but not during exercise in humans. 281(2), E365-74.
Brancaccio P, Lippi G, Maffulli N (2010) Clinical Chemistry and Laboratory Medicine. Biochemical markers of muscular damage. 48(6), 757-767.
Brancaccio P, Maffulli N, Limongelli F M (2007) British Medical Bulletin. Creatine kinase monitoring in sport medicine. 81-82:209-230.
Chaouloff F (1997) Medicine & Science in Sports & Exercise. Effects of acute physical exercise on central serotonergic systems. 29(1): 58-62.
Chen H. M, Muramoto K, Yamauchi F (1995) Agricultural and Food Chemistry. Structural analysis of antioxidative peptides from soybean β-conglycinin. 43: 574-578.
Davis J. M, Bailey S. P, Woods J. A, Galiano F. J, Hamilton M. T, Bartoli W P (1992) European Journal of Applied Physiology and Occupational Physiology. Effects of carbohydrate feedings on plasma free tryptophan and branched-chain amino acids during prolonged cycling. 65(6), 513-519.
Desmond K. O’Toole. (1999) Journal of Agricultural and Food Chemistry. Characteristics and use of okara, the soybean residue from soy milk productions –A review. 47(2), 363-371.
Dort J, Sirois A, Leblanc N, Côté C.H, Jacques H. (2012) Applied Physiology, Nutrition, and Metabolism. Beneficial effects of cod protein on skeletal muscle repair following injury. 37(3): 489-98.
Hsiu-Fen Yang, Tim K. Tso, Chen-Chih Huang, Wen-Nan Huang.(2015) Journal of Food and Nutrition Research. Okara Supplementation Relieves Fatigue and Muscle Damage that Occur during Exercise Training. 3(5), 352-356.
Katayama M, Wilson L A. (2008) Journal Food Sci. Utilization of okara, a byproduct from soymilk production, through the development of soy-based snack food. 73(3): S152-157.
Matsumoto K, Watanabe Y, Yokoyama S. (2007) Biosci Biotechnol Biochem. Okara, soybean residue, prevents obesity in a diet-induced murine obesity model. 71(3): 720-7.
Negro M, Giardina S, Marzani B, Marzatico F. (2008) The Journal of sports medicine and physical fitness. Branched-chain amino acid supplementation does not enhance athletic performance but affects muscle recovery and the immune system. 48(3): 347-351.
Nosaka K, Newton M. (2002) Medicine & Science in Sports & Exercise. Concentric or eccentric training effect on eccentric exercise-induced muscle damage. 34(1): 63-9.
Pedersen B. K, Ostrowski K, Rohde T, Bruunsgaard H. (1998) Canadian Journal of Physiology and Pharmacology. The cytokine response to strenuous exercise.76(5): 505-511.
Sola-Penna M (2008) International Union of Biochemistry and Molecular Biology. Metabolic regulation by lactate. 60(9): 605-608.
Strobel N. A, Peake J. M, Matsumoto A, Marsh S. A, Coombes J. S, Wadley G. D (2010) Medicine and Science in Sports and Exercise. Antioxidant Supplementation Reduces Skeletal Muscle Mitochondrial Biogenesis. 43(6): 1017-1024.
Tadano T, Yokoya K, Hirokawa R. (1981) Bull. Coll. Agr. Vet. Med. Studies on pneumatic.Drying of okara.38, 149-154.
Lemes F, Lima FM, de Almeida AP, Ramalho Ade F, Reis SR, Michelotto LF, Amaya-Farfán J, Carneiro EM, Boschero AC, Latorraca MQ, Veloso RV.(2014) International Journal of Food Sciences and Nutrition. Nutritional recovery with okara diet prevented hypercholesterolemia, hepatic steatosis and glucose intolerance. 65(6): 745-53.
Nakazato K, Ochi E, Waga T (2010) Molecular Nutrition & Food Research. Dietary apple polyphenols have preventive effects against lengthening contraction-induced muscle injuries. 54(3):364-72.
Trombold J. R, Barnes J. N, Critchley L, Coyle E. F. (2010) Medicine and Science in Sports and Exercise. Ellagitannin consumption improves strength recovery 2-3 d after eccentric exercise. 42(3):493-8.
Wu J, Ding X (2001) Journal of Agricultural and Food Chemistry. Hypotensive and physiological effect of angiotensin coverting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats. 49, 501-506.
Wigernæs I, Høstmark A. T, Strømme S. B, Kierulf P, Birkeland K (2001) European Journal of Applied Physiology. Active recovery and post-exercise white blood cell count, free fatty acids, and hormones in endurance athletes. 84(4):358-66