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研究生:黃詩愉
研究生(外文):Shin-yu Huang
論文名稱:納紅豆之安全性評估及對腸道菌相的影響
論文名稱(外文):Safety evaluation of natto-red bean and its effects on the intestinal microflora
指導教授:鍾雲琴
指導教授(外文):Yun-Chin Chung
學位類別:碩士
校院名稱:靜宜大學
系所名稱:食品營養研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:90
中文關鍵詞:腸道菌相安全性評估納紅豆
外文關鍵詞:intestinal microfloranatto-red beanSafety evaluation
相關次數:
  • 被引用被引用:3
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本研究室之前的研究結果顯示,紅豆經Bacillus subtilis發酵後,其50 % 乙醇萃取物之抗氧化能力及總多酚化合物較未發酵之紅豆佳;亦有研究指出,以B. subtilis發酵之黃豆具有改善腸道功能之效果。本實驗以B. subtilis發酵紅豆製成納紅豆,找尋其最佳發酵條件,並以SD (Sprague-Dawely) 大鼠作為動物模式探討其安全性及對動物腸道菌相之影響。納紅豆之最適發酵條件乃利用DPPH.(α,α-diphenyl-βpricrylhydrazyl)自由基清除能力及溶解血纖維蛋白能力評估之;納紅豆之安全性評估項目則包括Ames 試驗、微核試驗、14天單劑量毒性試驗 (急毒性) 及28天動物餵食毒性試驗 (亞毒性)。腸道菌相所分析的菌種包括Lactobacillus spp.、Bifidobacteria spp.、 Escherichia coli及Clostridium perfringens。結果顯示,納紅豆之最佳發酵條件為,以高雄五號做為基質並添加1 % 葡萄糖於30℃下發酵96小時。體外及體內基因毒性試驗 (Amse 試驗及微核試驗) 顯示納紅豆水萃液不具基因毒性及致突變性。動物餵食實驗結果證實納紅豆不具急毒性及亞急性,其無毒性劑量(no-observed-adverse-effect level, NOAEL) 超過1 g/kg BW。餵食納紅豆對於動物腸道中Lactobacillus spp.、Bifidobacteria spp. 、 E.coli及C. perfringens等菌量並無影響。 綜合上述,納紅豆對大鼠為安全性食品,但不具益菌生之特性。
Our previous study showed that the antioxidant capacity and total polyphenol content of 50% ethanol extract from red bean fermented by Bacillus subtilis were greater than that of from unfermented red beans. On the other hand, research showed that soybean fermented by B. subtilis could improve the intestinal functions. This study aimed to establish the optimum conditions for the B. subtilis-fermented red bean (called natto-red bean), and to evaluate the safety of natto-red bean and its effect on intestinal microflora using the Sprague-Dawely (SD) rats as a animal model. The α-diphenyl-βpricrylhydrazyl(DPPH)radicals scavenging effect and fibrinolytic activity were used as the index of fermentation result, and the Ames test, micronucleus test, 14-day feeding toxicity study (acute toxicity) and 28-day feeding toxicity study (subacute toxicity) were performed for safety evaluation. Intestinal microflora, such as Lactobacillus spp., Bifidobacteria spp., Escherichia coli, and Clostridium perfringens were determined as well. The results of this study showed that the optimum conditions for the production of Natto-red bean were to ferment Kaohsiung NO.5 red bean (Phaseolus radiatus L. var. aurea Prain) with addition of 1% Glucose using Bacillus subtilis at 30℃for 96 hr. The genotoxicity tests including in vitro and in vivo assays, Ames test and micronucleus test, respectively, showed the water extract of Natto-red bean had no genotoxicity. Animal feeding experiments, confirmed by 14-day feeding toxicity study and 28-day feeding toxicity study, showed that the natto-red bean had no toxicity, and the no-observed-adverse-effect level (NOAEL) was greater than 1 g / kg BW. The natto-red bean had no effect on intestinal microflora inculding Lactobacillus spp., Bifidobacteria spp., E. coli, and C. perfringens. In conclusion, the natto-red bean had no toxicity, and had no prebiotic effect either in rats.
壹、 文獻回顧 3
1. 紅豆的保健功效 3
1.1 簡介 3
1.2 紅豆生理活性 3
2. 納豆的保健功效 4
2.1 納豆菌 4
2.2 納豆之營養成分 5
2.3 納豆之生理活性 5
3. 致突變之機制探討 11
3.1 食品中之致突變物 11
3.1.1 天然毒素 12
3.1.2 食品添加物 13
3.1.3 加工或烹煮過程中產生之致突變物質 13
3.2 致突變作用機制 14
3.2.1 直接型致突變劑 14
3.2.2 間接型致突變劑 15
3.2.3 氧化型致突變劑 15
4. 食品安全性評估 16
4.1 微生物基因突變分析 (Ames test) 18
4.2 單一劑量口服毒性試驗 (Acute oral toxicity limit test) 22
4.3 28天餵食毒性試驗 (28-day feeding toxicity study) 22
4.4 微核試驗 22
5. 腸道菌與人體健康之關係 25
貳、 材 料 與 方 法 28
1. 實驗架構 28
2. 實驗材料 29
2.1 化學藥品 29
2.2 儀器 30
2.3 實驗動物 31
3. 實驗方法 31
3.1 納紅豆製備: 31
3.1.1 樣品: 31
3.1.2 菌種: 32
3.1.3 步驟: 32
3.2 最適發酵條件: 32
3.2.1 添加葡萄糖的影響 32
3.2.2 納紅豆水萃液製備: 32
3.2.3 DPPH.(Α,Α-DIPHENYL-ΒPRICRYLHYDRAZYL)清除能力: 33
3.2.4 血纖維蛋白平板試驗: 33
3.3 微生物基因毒性分析: 34
3.4 微核試驗: 40
3.5 單一劑量口服毒性試驗 (ACUTE ORAL TOXICITY LIMIT TEST): 41
3.6 28天餵食毒性試驗 (28-DAY FEEDING TOXICITY STUDY): 42
3.7 腸內菌相評估: 43
3.7.1 藥品配製: 43
3.7.2 實驗步驟: 46
3.7.3 計算公式如下: 47
參、 結果與討論 48
肆、 結論 77
伍、 參考文獻 78
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