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研究生:趙桂蓉
研究生(外文):Guei-Rung Chao
論文名稱:木耳類之抗氧化性質及其多醣理化分析
論文名稱(外文):Antioxidant Properties and Polysaccharide Physicochemical Analysis of Ear Mushrooms
指導教授:毛正倫毛正倫引用關係
指導教授(外文):Jeng-Leun Mau, Ph.D.
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:125
中文關鍵詞:木耳抗氧化性質多醣體
外文關鍵詞:Wood earsantioxidant propertiespolysaccharides
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木耳是我國普遍的食用菌之一。本省目前已商業化生產的木耳共有五種,包括黑耳、紅耳、斤耳、雪耳及白木耳。菇類之子實體和菌絲體可用來做為食品、調味料、特殊化學物質的生產(如水溶性多醣、維生素、有機酸、核苷酸和酵素)及醫療用物質生產(如抗生素、抗癌藥物和抗病藥物)。此外,菇類亦可提供抗氧化物質,但木耳類之抗氧化性質方面,在台灣並無學者研究木耳類之生理活性。故本研究則以木耳類子實體作為樣品,進行抗氧化性質評估並萃取多醣體,探討其多醣體之組成與結構特性。
木耳類甲醇萃取物抗氧化性質之測定,抗氧化活性方面在1,3-二乙基-2-硫巴比妥酸法於濃度1.0~5.0 mg/mL時,油脂氧化率38.6~74.6%,而共軛雙烯法在濃度5.0 mg/mL時,紅耳、斤耳及雪耳具較佳的抗氧化力。還原力測定在濃度5.0 mg/mL時雪耳>黑耳≈ 紅耳≈ 斤耳>白木耳。捕捉1,1-二苯基-2-苦味基團(DPPH·)法中除了白木耳外其他均具有較佳的捕捉能力。木耳類捕捉羥自由基(hydroxyl radical)之能力不佳但具有良好之亞鐵離子螯合能力。在天然抗氧化成分方面,木耳類之甲醇萃取物中發現含有抗壞血酸、生育醇和總酚類化合物,但沒有偵測到b-胡蘿蔔素的存在,而總抗氧化成分含量為黑耳15.69 mg/g、紅耳30.09 mg/g、斤耳27.83 mg/g、雪耳49.17 mg/g、白木耳31.70 mg/g。
木耳類子實體之多醣萃取物分析,水萃取及鹼萃取多醣之總糖含量在74~92%,多醣體之單糖組成以木糖、甘露糖及醛糖酸為主,且有些含有葡萄糖之中性單糖組成。木耳類水萃取及鹼萃取多醣體經膠體過濾層析後得知其皆含有大於2 ×106 Da之大分子。以核磁共振氫光譜判定多醣結構具有RCH3-、RCH2-、CH3-CO-、CH2O-官能基存在,且可能有b-D-葡聚糖的化學位移。紅外線光譜上具有糖類官能基之吸光特性,得知水萃取多醣體具有-OH基、C-H基、C=O基、C=C基、C-O基、-CH-O-CH-基之吸收帶出現,而鹼萃取多醣體之差異是缺少C=O基、C-O基之吸收帶。多醣之示差熱掃瞄熱分析儀測定,結果發現多醣粉末屬於吸熱反應,除白木耳的水萃取及鹼萃取多醣各有一明顯的波峰外其他木耳類皆沒有明顯的吸熱峰,而其理化特性分析需更進一步探討。由以上所得結果及X光繞射儀分析結晶構形,得知木耳類之多醣體皆可能有b-D-葡聚糖之結構,但其明確機制仍有待進一步純化大分子多醣體後,再加以探討。
綜合本研究之結果可知木耳類皆含有菇類主要成分之碳水化合物,且其甲醇萃取物皆具有良好的抗氧化性,而水萃取及鹼萃取兩種多醣皆含有大分子之多醣體,並以相似單糖組成存在於其子實體中,經光譜分析後皆得知其含有可能具生理活性之b-D-葡聚糖。
Wood ears are common edible fungi in China. Five kinds of wood ears are already commercially available in Taiwan, including black, red, jin, snow and silver ears. Mushroom fruit bodies and mycelia can be used as foods and food flavoring, to produce specifically chemical and medical substances, such as water soluble polysaccharides, vitamins, organic acids, nucleotides and enzymes, as well as to pharmaceuticals such as antibiotics, anticancer and antidisease drugs. In addition, mushrooms can supply antioxidant substances. However, so far no complete information is available for the antioxidant properties of wood ears in Taiwan. However, studies on the physiologically effects of wood ears have not been conducted.
This research used wood ears as samples to study the proximate composition, antioxidant properties of methanolic extracts, the monosaccharide constituents and the structural analysis of polysaccharides. Their structures were elucidated by spectroscopic analysis and comparison with known compounds.
For all methanolic extracts from ear mushrooms, the antioxidant activity in the 1,3-diethyl-2-thiobarbituric acid method were moderate (38.6~74.6%) at 1.0 to 5.0 mg/mL. Methanolic extracts for red, jin and snow ears showed excellent antioxidant activity in the conjugated diene method at 5.0 mg/mL. At 5.0 mg/mL, reducing powers in the descending order of sonw>black ≈ red ≈ jin>silver ears. The scavenging effect of methanolic extracts from ear mushrooms on 1,1-diphenyl-2-picryl hydrazyl radicals (DPPH·) was excellent except for that from silver ears. Ear mushrooms were not good scavengers for hydroxyl free radicals but good chelators for ferrous ions. Naturally occurring antioxidants, including ascorbic acid, tocopherols and total polyphenols, were found in methanolic extracts. However, b-carotene was not detected. Total antioxidant components were 15.69, 30.09, 27.83, 49.17, and 31.70 mg/g for black, red, jin, snow, and silver ears, respectively.
In ear mushrooms, the total sugar contents of water extract and alkaline extract polysaccharides were 74~92%. The compositions of neutral monosaccharide were the major have xylose, mannose, glucuronic acid and some ears have glucose. Ear mushrooms of water extract and alkaline extract polysaccharides was separated using gel filtration, and showed a molecular weight of about 2×106 Da. Their structure were elucidated by 1H-NMR spectra of water extract and alkaline extract polysaccharides showed that they contained RCH3, RCH2, CH3-CO- and CH2O-. That can be have a b-D-glucan chain. IR spectroscopy showed that water extract polysaccharides contained OH, C-H, C=O, C=C, C-O, CH-O-CH-, but that alkaline extract polysaccharides not have C=O, C-O.
The thermal behaviors of water extract and alkaline extract polysaccharides were examined by DSC, except for silver ear showed one endotherm but the anther not have. It could be study their physiological activities, advance analysis. The research and X-ray can be have a b-D-glucan chain. For the application in healthy food, further research on the mechanism of polysaccharide properties is in progress.
表次……………………………………………………………………ix
圖次……………………………………………………………….……x
前言…………………………………………………………………….1
文獻整理…………………………………………………………….3
一、木耳之介紹……...……………….……………………………….….3
二、菇類的機能性………...………………..………………………….…6
三、抗氧化性質……………………………...………………………….9
四、菇類多醣…………..………………………………...……………...21
材料與方法…………………………………………………..……27
一、實驗材料……………...………………………………….…………27
1. 木耳….…………..…..………………………………………..27
2. 藥品…………………………………….……………………..27
二、實驗方法………..………………………………………….……….28
(一)木耳類甲醇萃取物之抗氧化性質分析….…………….….….28
1.木耳類子實體粉末製備…………………………...……….28
2.水分之測定………………………………………...……….28
3.樣品萃取……………………………………………………30
4. 抗氧化力之測定……………………………………………30
(1) 1,3-二乙基-2-硫巴比妥酸(DETBA)法之測定……..…30
(2) 共軛雙烯(Conjugated diene)法之測定………….……30
5.還原力之測定………………………………….….…….….31
6. 捕捉1,1-二苯基-2-苦味基團(DPPH·)能力之測定……31
7. 捕捉羥自由基能力之測定.….….….….……..…..…..…….32
8. 螯合亞鐵離子能力之測定…………………………………32
(二)木耳類甲醇萃取物之抗氧化成分分析………………….…...32
1.總酚類化合物測定…………………………………………32
2.抗壞血酸含量測定…………………………………………33
3.b-胡蘿蔔素測定……………………………………………33
4.生育醇測定…………………………………………………34
(三)木耳類多醣組成份之分析………….……...…………………34
1. 木耳類子實體多醣之製備………………...……………….34
2. 木耳類多醣組成分之分析………………………………….35
a. 水分之測定……………………………………………….35
b. 灰分之測定……………………………………………….35
c. 總醣含量之測定…………………….……………………35
d. 微量蛋白之測定………………….………………………36
e. 含氮量之測定…………………………………………….36
f. 醛醣酸含量之測定……………………………………….36
g. 中性單糖組成之測定……………………….……………37
(四)木耳類多醣理化之分析……….……….....…………………..37
1. 分子量之測定……………………………………………….37
2. 結構分析…………………………………………………….38
a. 核磁共振-氫光譜(1H-NMR)之分析……………...……..38
b. 紅外線(IR)光譜之分析…………….……………………39
c. X光繞射(X-ray)之分析…………………....…………..39
d. 示差掃描熱分析……………………..…………….……..39
三、統計分析………………………………………...………………….39
結果與討論………………………………………………………..40
一、木耳類甲醇萃取物之抗氧化性質……………..………...….…..…40
1. 木耳類之含水率測定…………………………………..………40
2. 木耳類甲醇萃取物的萃取率…………………………..………40
3. 木耳類甲醇萃取物之抗氧化力………………………..………40
4. 木耳類甲醇萃取物之還原力……………………………..……49
5. 木耳類甲醇萃取物捕捉1,1-二苯基-2-苦味基團之能力…..52
6. 木耳類甲醇萃取物捕捉羥自由基之能力……………….….…55
7. 木耳類甲醇萃取物螯合亞鐵離子之能力………………..……56
二、 木耳類甲醇萃取物之抗氧化成分分析……….…….……..……..62
三、 木耳類多醣組成份之分析……………….…..……….…..………65
1. 木耳類多醣體萃取率………...…………………….…………..65
2. 木耳類多醣體之成分分析…...……………….………………..67
3. 木耳類多醣之元素分析………………...…………………...…67
4. 木耳類中性單糖組成分分析………………….…………….…71
四、 木耳類多醣分子量測定及其理化分析...…...……….…...……..74
結論……………………………………………………...…..…..…..109
參考文獻……………………………………...…………...………110
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