臺灣博碩士論文加值系統

紅棗( Zizyphus jujuba )與胖大海( Sterculia lychnophora )為台灣常見的中藥材，常利用於藥膳食譜當中。本實驗的目的是探討不同溶液 ( 酸溶液、鹼溶液 & 水 ) 萃取粗多醣，在化學組成、抗氧化力及黏度特性的變化。並將粗多醣進一步經酸水解且利用膠體過濾層析的方法進行分離，以了解不同酸水解程度的紅棗與胖大海多醣之化學組成及結構上的差異，以及其多醣對腸道菌增生的影響。實驗結果顯示，以酸溶液及鹼溶液的粗多醣萃取率較高；利用鹼溶液萃取的紅棗與胖大海多醣具有較高的醣醛酸含量與較佳的抗氧化能力；粗多醣的黏度則以水溶液萃取的紅棗及胖大海多醣最高。在酸水解過後經膠體過濾層析可得到兩個分子量分布，取小分子多醣與未水解多醣進行化學組成、結構與腸道菌增生影響的比較。結果顯示，多醣分子量會隨著水解時間增加而有降低的趨勢；在化學組成比例的變化上，總醣含量皆因經過膠體過濾層析劃分後有顯著提高的趨勢，而醣醛酸含量則隨著水解時間增加而有降低的趨勢；針對腸道菌增生的影響，添加紅棗與胖大海的水解多醣，以水解2 hr及4 hr的小分子多醣濃度1mg / mL時，皆對Bifidobacteriun bifidum有增生的效果，其中以胖大海小分子多醣效果較佳。水解2 hr及4 hr的小分子紅棗多醣對Propionibacterium freudenreichii也有增生的效果，而胖大海則不明顯，推測兩者的差異可能與水解後多醣的醣組成及結構有關。

Zizyphus jujuba ( ZJ ) and Sterculia lychnophora ( SL ) are the traditional Chinese herbs using in the medicated food. The purposes of this study are to investipate the physicochemical properties and antioxidant capacity of polysaccharides of ZJ and SL. Furthermore, the effect of acid hydrolyzed polysaccharides on the growth of internal microflora. The crude polysaccharides were acid hydrolyzed by using diluted hydrochloride solution (2M) for 6hr. The hydrolytes were fractionated by using gel permeation chromatography (GPC) to obtain various molecular weight of polysaccharides. The results indicated that the extraction of acidic and basic solutions obtained higher yield of crude polysaccharides than hot water extraction. The crude polysaccharides from basic solution had the more uronic acid content and the better antioxidant capacity than the others. In the viscosity of crude polysaccharide solutions, water extracts indicated the highest viscosity among three crude polysaccharides in both ZJ and SL. Two major molecular weight distributions were observed as the acid hydrolytes were fractionated by using GPC. The total carbohydrate of hydrolytes was increased after GPC fractionation. However, the content of uronic acid in the acid hydrolyzed polysaccharides was decreased with progress of hydrolysis time. The addition of acid hydrolytes (both 2hr and 4hr) significantly increased the growth of Bifidobacteriun bifidum as the concentration of 1mg / mL was added, particularly in the fraction of low molecular weight of SL. Similarly, the acid hydrolytes (both 2hr and 4hr) of ZJ also significantly increased the growth of propionibacterium freudenreichii, but no significant effect was observed as SL hydrolytes was added. The differences between hydrolytes of SJ and SW could be caused by the different molecular structure of polysaccharide

摘要 I
Abstract II
目錄 IV
表目錄 VII
圖目錄 IX
第一章 前言 1
第二章 文獻回顧 3
ㄧ、紅棗 3
二、胖大海 4
三、多醣 5
四、萃取法 7
五、多醣的純化 ( Purification ) 10
六、劃分 ( Fractionation ) 16
七、多醣的流變特性 19
八、抗氧化能力 21
( 一 ) 自由基與活性氧 21
( 二 ) 抗氧化作用的機制 22
( 三 ) 多醣的抗氧化 24
第三章 材料與方法 31
一、實驗架構 31
二、材料 32
三、粗多醣製備 32
壹、紅棗與胖大海 35
一、一般成分分析 35
(一) 水分測定 35
(一) 粗蛋白測定 35
(二) 粗脂肪測定 35
(三) 灰份測定 35
(四) 粗纖維含量 35
(五) 無氮抽出物 36
貳、粗多醣 36
一、化學組成分析 36
(一) 總醣 (Dubois et al., 1956) 36
(二) 蛋白質 (Smith et al., 1985) 37
(三) 醣醛酸(Blumenkrantz & Asboe-Hansen, 1973) 39
(四) 中性單醣 (Kerherve et al., 1995) 41
二、抗氧化特性 43
(一) DPPH自由基清除能力 43
(二) 還原力 46
（三） 螯合亞鐵離子的能力 47
三、黏度特性 49
叁、酸水解多醣 50
一、化學組成分析 50
(一) 總醣 50
(二) 蛋白質 50
(三) 醣醛酸 50
(四) 中性單醣 50
二、結構和分子量測定 51
(一) 結構分析 51
(二) 分子量測定 52
三、腸道菌群增生的影響 53
肆、統計分析 57
第四章 結果與討論 58
一、基本成分分析 58
二、 多醣化學組成 63
(一) 總醣含量 63
(二)蛋白質含量 63
(三)醣醛酸含量 64
三、 抗氧化特性 70
(一) DPPH自由基清除能力 70
(二) 還原力 74
(三) 亞鐵離子螯合能力 78
四、濃度與溫度對黏度的影響 84
五、酸水解後多醣化學組成 88
(一) 總醣含量 88
(二)蛋白質含量 88
(三)醣醛酸含量 89
(四)中性單醣組成 92
六、多醣結構與分子量 95
(一)分子量大小 95
(二)多醣官能基 96
七、小分子多醣對腸道菌生長的影響 102
第五章、結論 115
參考文獻 116

丁克祥︰SOD生物醫學淺論。藝軒圖書出版社。台北，台灣（1996）。
王煥華：中華食物養生大全。廣東旅遊出版社( 2009 )。
王伯徹、陳啟楨、華傑：食藥用菇類的培養與應用。食品工業發展研究所。56-57（1998）。
行政院衛生署:台灣地區食品營養成份資料庫。行政院衛生署。台北，台灣 ( 1998 )。
吳豔、顧小紅、Steve W. Cui、湯堅。胖大海酸性多醣的分離純化及初步結構研究。食品研究與開發，7:95-98 (2006)。
李全順：β-胡蘿蔔素和維生素 E 對大鼠抗輻射作用的實驗研究。衛生職業教育。21（3）：135（2003）。
李向志：木耳、白木耳豐產技術。五洲出版社。台中。台灣（1989）。
沙玉清：泥沙運動學引論，陝西科學技術出版社。（1996）。
周鄭坤、周文明、賀明宇 & 吳小青。胖大海水溶性多醣的提取。西北林學院學報，21：129-131 ( 2006 )。
卓秋萍：枸杞多醣的抗氧化能力與益菌生功能的探討。靜宜大學食品營養研究所碩士論文。台中，台灣( 2008 )。
高曉燕、蘇又凡、李發美：牛膝中胡蘿蔔甙對成骨樣細胞的促進增殖作用　　　　　　　及其含量測定。承德醫學院學報（2003）。
徐美如：中草藥水萃物抗氧化及誘導肝癌細胞凋亡之研究。國立中興大學食品科學系碩士論文。台中，台灣（2006）。
馬文濤：維生素C對應激大鼠腦內超氧化物歧化酶含量的影響。中國心理衛生雜誌，12：809（2002）。
張為憲、李敏雄、呂政義、張永和、陳昭雄、孫璐西、陳怡宏、張基郁、顏國卿、林志城、林慶文。食品化學。華香圓出版社。台北，台灣（1995）。
陳倩琪、彭宜融、廖麗玲。基因體研究對乳酸菌機能保健品之產業發展潛力。食品生技，11：52-59 ( 2007 )。
陳國梁、張金文，陳宗禮、張向前:紅棗多醣提取分離工藝的優化。食品科學，27:531-534 (2006)。
黃惠宇：腸道系統守護神-乳酸菌。科學期刊，37(2)：104-107 ( 2007 )。
楊雲、李振國、孟江、魏江。大棗多醣的分離、純化及分子量的測定。 世界科學技術，5(3)︰53-55 ( 2003 )。
趙士豪，路新利，郭會燦：維生素E保健作用的研究進展。山西食品工業，1：32（2005）。
樊誠哲，陽明曾：木耳、銀耳、草菇。五洲出版社。台中。台灣（1986）。
劉宜璠：探討紅棗萃取液對腸道菌增生能力及抗氧化和抗腫瘤活性的變化。 靜宜大學食品營養研究所碩士論文。台中，台灣( 2008 )。
賴慶亮譯，水野卓、川合正允：菇類化學.生化學。國立編譯館，台北，台灣（1997）。
羅凱、胡延章、黃小雲、楊東升︰紅棗中多醣提取及純化工藝的研究。安徽農業科學，34(22)︰5962-5963（2006）。
A.A.C.C. 2000. Approved methods of the A.A.C.C. 8 th ed. American Association of Cereal Chemists, St. Paul, MN.
Ballard, T. S., Mallikarjunan, P., Zhou, K. Q., & O’Keefe, S. ( 2010 ). Microwave-assisted extraction of phenolic antioxidant compounds from peanut skins. Food Chemistry, 120, 1185–1192.
Bisicchia R., Sozzi T., Bertelli M., Bonora R. & Zunin C. ( 1984 ). Bifidobacteria colonization in diversely-fed newborn infants. Pediatria Medicae Chirurgica 6(3), 395-400
Bitter, T., & Muir, H. M. ( 1962 ). A modified uronic acid carbazolee reaction. Analytical Biochemistry, 4, 330-334.
Blumenkrantz, N., & Asboe-Hansen, G. ( 1973 ). New method for quantitative determination of uronic acid. Analytical Biochemistry, 54, 484-489.
Bohn, J. A. & Bemiller, J. N. ( 1995 ). ( 1→3 )-β-Glucans as biological response modifiers: a review of structure-functional activity relationships. Carbohydrate Polymers, 28,3-14.
Bursal, E., & Gülçin, İ. ( 2011 ). Polyphenol contents and in vitro antioxidant activities of lyophilised aqueous extract of kiwifruit (Actinidia deliciosa). Food Research International, 44, (2011) 1482–1489.
Cadenas, E. ( 1989 ). Biochemistry of oxygen toxicity. Annual Review of Biochemistry, 58, 79-110.
Chang, S. C., Hsu, B. Y., Chen, B. H.( 2010 ). Structural characterization of polysaccharides from Zizyphus jujuba and evaluation of antioxidant activity. Biological Macromolecules, 47, 445-453.
Chen, Y., Cheng, P., Lin, C., Liao, H., Chen, Y., Chen, C. (2008). Polysaccharides from Antrodia camphorata mycelia extracts possess immunomodulatory activity and inhibits infection of Schistosoma mansoni. International Immunopharmacology, 8, 458–467.
Decker, E. A., Welch, B. ( 1990 ). Role of ferritin as a lipid oxidation catalyst in muscle food. Journal of Agricultural and Food Chemistry, 38, 674-677.
Ding, X., Tang, J., Cao, M., Guo, C. X., Zhang, X., Zhong, J., Zhang, J., Sun, Q., Feng, S., Yang, Z. R. & Zhao, J. ( 2010 ). Structure elucidation and antioxidant activity of a novel polysaccharide isolated from Tricholoma matsutake. Biological Macromolecules, 47, 271-275.
Dubois, M., Gilles, K. A., Hamition, J. K., Robers, A. P., & Smith, F. ( 1956 ). Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28, 350-356.
Dziezak, J. D. ( 1986 ). Preservatives : antioxidant. Food Technology, 40, 94-102.
Ge,Y.,Duan,Y., Fang,G., Zhang,Y., &Wang, S ( 2009 ).Polysaccharides from fruit calyx of Physalis alkekengi var. francheti: Isolation, purification, structural features and antioxidants. Carbohydrate Polymers, 77, 188-193.
Gibson, G. R., ＆ Roberforid, M. B. ( 1995 ). Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. Journal of Nutrition, 125, 1401-1412.
Gibson, G. R. (1999). Dietary modulation of the human gut microflora using the prebiotics oligofructose and Inulin. American Society for Nutritional Sciences, 129, 1438-1441.
Gilliland, S.E., Nelson, C.R. and Maxwell, C. ( 1985 ). Assimilation of cholesterol by Lactobacillus acidophilus. Applied and Environmental Microbiology . 49, 377-381
Halliwell, B., Murcia, M. A., Chirico, S., ＆ Aruoma, O. I. ( 1995 ). Free radicals and antioxidants in food and in vivo: what they do and how to work. Critical Reviews in Food Science and Nutrition, 35(1-2), 7-20.
He, X. Y., Xu, J. F., Wang, Q. S., & Chen, Y. Y. (2010). Ultrasonic extraction and determination of polysaccharide in Radix Angelica sinensis. China Pharmaceuticals, 19, 34-35.
Heo, S. J., Park, E. J., Lee, K. W., & Jeon, Y. J. (2005). Antioxidant activities of enzymatic extracts from brown seaweeds. Bioresource Technology, 96, 1613–1623.
Hung, C. F., Hsu, B. Y., Chang, S. C., Chen, B. H. ( 2012 ). Antiproliferation of melanoma cells by polysaccharide isolated from Zizyphus jujube. Nutrition, 28, 98–105
Jacob, R. A. ( 1995 ). The integrated antioxidant system. Nutrition Research, 15, 755-766.
Jiang, J., Guo, Y. J., & Niu, A. J. ( 2009 ). Extraction, characterization of Angelica sinensis polysaccharides and modulatory effect of the polysaccharides and Tai Chi exercise on oxidative injury in middle-aged women subjects. Carbohydrate Polymers, 77, 384-388.
Karbownik, M., ＆ Lewinski, A. ( 2003 ). The role of oxidative stress in physiological and pathological processes in the thyroid gland; possible involvement in pineal-thyroid interactions. Neuroendocrinology Letters, 24, 293-303.
Kerherve, P., Charriere, B., & Gradel, F. ( 1995 ). Determination of marine monosaccharides by high-pH anion exchange chromatography with pulsed amperometric detection. Journal of Chromatography, 718, 283-289.
Lai, L.S., Chou, S.T., Chao, W.W. (2001). Studies on antioxidative activities of hsian-tsao (Mesona procumbens Hemsl) leaf gum. Journal of Agricultural and Food Chemistry, 49, 963-968.
Lee, J. S., Kwon, J. S., Yun, J. S., Pahk, J. W., Shin, W. C. & Lee, E. K. H. ( 2010 ). Structural characterization of immunostimulating polysaccharide. from cultured mycelia of Cordyceps militaris. Carbohydrate Polymers, 80, 1011-1017.
Li, J., Ding, S. D., Ding, X. L. (2007). Optimization of the ultrasonically assisted extraction of polysaccharides from Zizyphus jujuba cv. Jinsixiaozao. Food Engineering. 80, 176-183.
Li, J., Fan, L., Ding, S. ( 2011). Isolation, purification and structure of a new water-soluble polysaccharide from Zizyphus jujube cv. Jinsixiaozao. Carbohydrate Polymers. 83, 477-482.
Li, S. P., & Tsim, K. W. K. (2004). The biological and pharmacological properties of Cordyceps sinensis, a traditional Chinese medicine that has broad clinical applications. In L. Packer, C. N. Ong, & B. Halliwell (Eds.), Herbal and traditional medicine: Molecular aspects of health (pp. 657–683). New York: Marcel Dekker.
Lia, G., Kimd, D. H., Kimb, T. D., Parka, B. J., Parka, H. D., Parkb, J. I., (2004). Protein-bound polysaccharide from Phellinus linteus induces G2/M phase arrest and apoptosis in SW480 human colon cancer cells. Cancer Letters, 216, 175–181.
Liu, P. J., Hsieh, W. T., Huang, S. H., Liao, H. F., & Chiang, B. H. (2010). Hematopoietic effect of water-soluble polysaccharides from Angelica sinensis on mice with acute blood loss. Experimental Hematology, 38, 437-445.
Ma, Z., Wang, J., Zhang, L., Zhang, Y. & Ding, K. ( 2010 ). Evaluation of water soluble b-D-glucan from Auricularia auricular-judae as potential anti-tumor agent. Carbohydrate Polymers, 80, 977-983.
Mazumder, S., Morvan, C., Thakur, S., & Ray, B. J. ( 2004 ). Cell Wall Polysaccharides from Chalkumra (Benincasa hispida) Fruit. Part I. Isolation and Characterization of Pectins. Journal of Agriculture and Food Chemistry, 52, 3556–3562.
Moore, W.E.C & Holdeman, L.V. ( 1974 ). Human fecal flora:the normal flora of 20 japancess-Hawaiians. Applied Microbiology. 27, 961.
Nie, S. P., & Xie, M. Y. (2011). A review on the isolation and structure of tea polysaccharides and their bioactivities. Food Hydrocolloids, 25, 144-149.
Okazaki, M., Koda, H., Izumi, R., Fujikawa, S. & Matsumoto, N. ( 1991 ). In vitro digestibility and in vivo utilization of xylobiose. Journal of Japan Society Nutrition and Food Science, 44, 41-44.
Prasad, K. N., Hao, J., Shi, J., Liu, T., Li, J., Wei, X., & Qiu, S. ( 2009 ). Antioxidant and anticancer activities of high pressure-assisted extract of longan (Dimocarpus longan Lour.) fruit pericarp. Innovative Food Science and Emerging Technologies, 10, 413-419.
Richards, D. M., Dean, R. T., & Jessup, W. ( 1988 ). Membrane protein are critical targets in free radical mediated cytolysis. Biochinica et Biophysica Acta, 946, 281-288.
Saarela M., Mogensen G., Fondèn R., Mättö J. and Tinna M.S. ( 2000 ). Probiotic bacteria: safety, functional and technological properties. Journal of Biotechnol. 84, 197-215.
Salminen, S. & Ouwehand, A. C. ( 2004 ). Probiotics. Biochemistry and food Chemistry, 18, 299-313.
Smith, P. K., Kroho R. I. Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., & Klenk, D. C. ( 1985 ). Measurement of protein using bicinchoninic acid. Analytical Biochemistry, 150, 76-85.
Somboonpanyakul, P., Wang, Q., Cui, W., Barbut, B., ＆ Jantawat, P. ( 2006 ). Malva nut gum. (Part I): Extraction and physicochemical characterization. Carbohydrate Polymers, 64, 247-253
Sun, Y. X., Li, T. & Liu, J. ( 2010a ). Structural characterization and hydroxyl radicals scavenging capacity of a polysaccharide from the fruiting bodies of Auricularia polytricha. Carbohydrate Polymers, 80, 377-380.
Sun, Y. X., & Liu, J. C. (2008). Structural characterization of a water-soluble polysaccharide from the Roots of Codonopsis pilosula and its immunity activity. International Journal of Biological Macromolecules, 43, 279–282.
Sun, Z., W., Zhang, L., X., Zhang, B., & Niu, T., G. (2010b). Structural characterisation and antioxidant properties of polysaccharides from the fruiting bodies of Russula virescens. Food Chemistry, 118, 675-680
Tannock, G. W. ( 1990 ). The microbiology of lacbotacilli inhabiting the gastrointestinal tract. In: advances in microbial ecology, Marshall, K. C.,Ed. ( pp. 147-171). New York, Plenum Press .
Valko, M., Lebifritz, D., Moncol, J., Cronin, M., Mazur, M., & Telser, J. ( 2007 ). Free radicals and antioxidants in normal physiological functions and human disease. The International Journal of Biochemistry and Cell Biology, 39, 44-84.
Wang, K. P., Zeng, F., Liu, J. Y., Guo, D., & Zhang, Y. ( 2011 ). Inhibitory effect of polysaccharides isolated from Angelica sinensis on hepcidin expression. Journal of Ethnopharmacology, 134, 944-948.
Wang, Q., & Fang, Y. ( 2002 ). Analysis of sugers in traditional Chinese drugs. Journal of chromatography B, 812, 219-231
Wang, Y., Wei, X. & Jin, Z. ( 2009 ). Structure analysis of a neutral polysaccharide isolated from green tea. Food Research International. 42, 739-745.
Whistler R L. ( 1973 ). Factors influencing gum costs and applications. In：Whistler R L. Industrial Gums, Polysaccharides and Their Derivatives. ( pp.6-7 ). New York, Academic Press.
Wu, Y., Cui, S. W., Tang, J. & Gu, X. ( 2007a ). Optimization of extraction process of crude polysaccharides from boat-fruited sterculia seeds by response surface methodology. Food Chemistry, 105, 1599-1605.
Wu, Y., Cui, S. W., Tang, J., Wang, Q. & Gu, X. ( 2007b ). Preparation, partial characterization and bioactivity of water-soluble polysaccharides from boat-fruited sterculia seed. Carbohydrate Polymers, 70, 437-443.
Yamaguchi, T., Takamura, H., Mattoba, T., & Terao, J. ( 1998 ). HPLC method for evaluation of the free radical-scavenging activity of foods by using 1,1-diphenyl-2-picrylhydrazyl. Bioscience, Biotechnology, and Biochemistry, 62(6), 1201-1204.
Yan, J. K., Li, L., Wang, Z. M., Lenug, P. H., Wang, W. Q. & Wu, J. Y. ( 2009 ). Acidic degradation and enhanced antioxidant activities of exopolysaccharides from Cordyceps sinensis mycelial culture. Food Chemistry, 117, 641–646.
Yang, X., Zhao, Y., Li, G., Wang, Z., & Lv, Y. ( 2008 ). Chemical composition and immuno-stimulating properties of polysaccharide biological response modifier isolated from Radix Angelica sinensis. Food Chemistry, 106, 269-276.
Yen, G. C., & Chen, H. Y. ( 1995 ). Antioxidant activity of various tea extracts in relation to their antimutagenicity. Journal of Agricultural and Food Chemitry, 43,27-32
Yuan, J. F., Zhang, Z. Q., Fan, Z. C., ＆ Tang, J. X. ( 2008 ). Antioxidant effects and cytotoxicity of three purified polysaccharides from Ligusticum chuanxiong Hort. Carbohydrate Polymers, 4, 822-827.
Yuan, F., Yu, R., Yin, Y., Shen, J., Dong, Q., Zhong, L. & Song, L. ( 2010 ). Structure characterization and antioxidant activity of a novel polysaccharide isolated from Ginkgo biloba. Biological Macromolecules, 46, 436-439.
Zhang, M., Cui, S. W.,Cheung, P. C. K., & Wang, Q.( 2007 ). Antitumor polysaccharides from mushrooms: a review on their isolation process, structural characteristics and antitumor activity. Trends in Food Science and Technology, 18, 4-19.