臺灣博碩士論文加值系統

中藥黃耆為豆科植物黃耆Astragalus membranaceus (Fisch.)Bunge的乾燥根，具有抗病毒、抗腫瘤、免疫調節、促進機體代謝等功效，多醣為其主要活性成分之ㄧ。本研究之目的為探討黃耆水溶性多醣之特徵與組成。結果顯示，黃耆水溶性多醣中的類澱粉多醣，α-(1,4；1,6)-D-glucans，佔了黃耆水溶性多醣之87.9％，利用碘呈色反應呈現藍紫色，可利用α-amylase、amyloglucosidase兩澱粉酵素水解並去除。非澱粉多醣為不被α-amylase、amyloglucosidase水解之部分，佔了水溶性多醣之12.1％，可利用四級銨鹽(cetrl trimethyl ammonium bromide, CTAB)選擇性錯合沉澱反應劃分成中性與酸性多醣區分。非澱粉多醣含有42.1％之醛醣酸，中性醣組成主要有阿拉伯糖、半乳糖，葡萄糖、甘露糖與岩藻糖，相對莫耳百分比為47.5：33.6：8.9：8.3：1.7％。進一步利用四級銨鹽（CTAB，cetyl trimethyl ammonium bromide）帶正電荷之特性選擇性錯合沉澱具高醛糖酸含量之非澱粉多醣，當中有21.3％可被四級銨鹽錯合沉澱，稱之為酸性多醣區分，不被四級銨鹽錯合沉澱者稱之為中性多醣區分。酸性多醣區分其單醣組成主要有阿拉伯糖（29.8％）、半乳糖（21.3％）、半乳醣醛酸（37.7％）及葡萄醣醛酸（2.7％），重量平均分子量為130kDa。中性多醣區分之主要單醣組成有阿拉伯糖（54.4％）、半乳糖（34.3％）、甘露糖（9.7％）與葡萄糖（1.5％），重量平均分子量為189kDa，為一廣分佈之多醣。利用陰離子交換樹脂可將中性多醣區分劃分成三區塊。中性與酸性多醣區分利用periodate-thiobarbituric acid method和β-D-glucosyl Yariv antigen皆呈現正反應，證明了KDO (2-keto-3-deoxy-D-manno-octulosonic acid)和arabinogalactan Type Ⅱ之存在。結果顯示經此一流程可獲得高醛醣酸含量、具有果膠多醣特徵之黃耆多醣。

The dried root of Astragalus membranaceus (Fisch.) Bunge, also called Astragali radix, is an important traditional Chinese medicine. It has the functions of anti-virus, anti-tumor, and immuno-modulating activities. Polysaccharide is one of the active components of this herb. The focus of this study is to investigate the characteristics of water-soluble polysaccharides from A. membranaceus. The α-(1,4; 1,6)-D-glucans, the starch-like polysaccharides showing blue color under iodine-staining, in the hot-water extracted crude polysaccharides were digested and removed by α-amylase and amyloglucosidase digestion. The non-starch polysaccharides, the non-digestible portion, were fractionated by using selective precipitation with cetyl trimethyl ammonium bromide, CTAB, into acidic and neutral polysaccharides fractions. The α-(1,4; 1,6)-D-glucans was found to be the most abundant component in the crude polysaccharides and its content was 87.9％. The non-starch polysaccharides contained 42.1% of uronic acids and the sugar compositions of the neutral portion were arabinose, galactose, glucose, mannose and fucose in the molar ratio of 47.5: 33.6: 8.9: 8.3:1.7 %. The CTAB could react with the polysaccharides with high ratio of uronic acids to form precipitates and successfully separated non-starch polysaccharides into acidic and neutral polysaccharides fractions with ratio of 21.3% and 78.7%, respectively. The acidic polysaccharides fraction consisted of arabinose (29.8%), galactose (21.3%), galacturonic acid (37.7%), and glucuronic acid (2.7%). The weight-average molecular weight of acidic polysaccharides fraction was 130 kDa. The neutral polysaccharides fraction consisted of arabinose (54.4%), galactose (34.3%), mannose (9.7%) and glucose (1.5%). The weight-average molecular weight of neutral polysaccharides fraction was 189 kDa with wide range of distribution. This fraction can be further divided into three sub-fractions by anion- exchange chromatography. Both acidic and neutral polysaccharide fractions showed positive reaction with periodate-thiobarbituric acid and β-D-glucosyl Yariv antigen assays indicated the existing of KDO (2-keto-3-deoxy-D-manno-octulosonic acid) and branched (1,3; 1,6)-arabinogalactans. These results indicated existing of hairy domains of pectic polysaccharides in the acidic polysaccharides and low-uronic acid containing polysaccharides in neutral polysaccharides fraction of A. membranaceus.

中文摘要.....................................Ι
abstract.........................................Ⅱ
總目錄.........................................Ⅲ
圖目錄.........................................Ⅵ
表目錄.........................................Ⅶ
壹、前言.................................................1
貳、文獻回顧.............................................................2
一、黃耆介紹..................................................2
(一) 型態與分佈.............................................2
(二) 栽培環境與生理特性................................................2
(三) 活性成分...............................................................3
1. 多醣類..........................................3
2. 皂苷類..........................................4
(四) 生理活性.......................................................4
1. 抗病毒.................................................................5
2. 抗腫瘤..................................5
3. 清除自由基能力..................5
4. 對免疫系統之作用..............5
5. 造血及心血管系統保護作用..............6
6. 肝臟保護作用........................................................................7
7. 促進機體代謝.................................7
8. 其他.....................................................................7
二、果膠多醣（pectic polysaccharide）...............................8
(一) Homogalacturonan（HG）......................................8
(二) Xylogalacturonan（XGA）......................................8
(三) RhamnogalacturonanⅠ（RGⅠ）.................................9
(四) RhamnogalacturonanⅡ（RGⅡ）...................................9
三、具活性成分之果膠多醣...........................................14
(一) Arabinogalactan TypeⅡ（AGⅡ）............................14
(二) RhamnogalacturonanⅠ（RGⅠ）................................15
參、材料與方法...................................................19
一、實驗材料.............................................................19
二、實驗藥品與試劑..........................................................19
(一) 化學試劑................................................19
(二) 標準品....................................................19
(三) 酵素.....................................20
三、樣品製備方式...............................................20
(一) 熱水萃取物之製備........................................20
(二) 水溶性粗多醣之製備.....................................20
(三) 非澱粉多醣...................................20
(四) 陰離子交換樹脂分析水溶性粗多醣成分.....................20
(五) CTAB沉澱酸性多醣區分與中性多醣區分.....................21
四、分析方法................................................21
(一) 水分含量測定...............................................21
(二) 固形物含量測定..........................21
(三) 碳水化合物含量測定............................21
(四) 醛醣酸含量測定................................22
(五) KDO含量測定........................................22
(六) 蛋白質含量測定................................22
(七) 單醣組成測定（TFA hydrolysis）.......................22
(八) 單醣組成測定（methanolysis）.................................23
(九) 分子量測定....................................................24
(十) β-D-glucosyl Yariv antigen test...........................24
五、實驗流程.........................................................26
肆、結果與討論..........................................27
一、水萃物與水溶性粗多醣之組成分析...................27
(一) 黃耆熱水萃取物與水溶性粗多醣組成分析..............27
(二) 水溶性粗多醣分子量分佈........................28
(三) 水溶性粗多醣組成分析.................................29
(四) 水溶性粗多醣各成分之單醣組成分析......................30
二、水溶性粗多醣之劃分..........................................33
(一) α-(1,4；1,6)-D-葡聚醣之水解及非澱粉多醣之分離...................33
(二) 中性及酸性多醣區分之劃分..................................37
1. 中性多醣區分.......................................39
甲、多醣組成分析...............................39
乙、分子量分佈................................................40
丙、單醣組成分析.......................................45
2. 酸性多醣區分...................................................46
甲、多醣組成分析...................................................................46
乙、分子量分佈.......................................................................47
丙、單醣組成分析...................................................................48
三、多醣結構偵測....................................................49
(一) KDO含量測定..........................................49
(二) β-D-glucosyl Yariv antigen test....................50
伍、結論..........................................52
陸、參考文獻..................................53
柒、附錄..........................................62

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