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研究生:林蔚任
研究生(外文):Wei-Ren Lin
論文名稱:由草莓粕製備鞣花酸濃縮物之方法及鞣花酸濃縮物之安全性評估
論文名稱(外文):Preparation of ellagic acid concentrate from strawberry pomaces and safety evaluation thereof
指導教授:蘇南維蘇南維引用關係
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:農業化學研究所
學門:農業科學學門
學類:農業化學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:76
中文關鍵詞:草莓粕鞣花酸鞣花酸濃縮物安氏試驗法L5178Y TK分析法
外文關鍵詞:strawberry pomacesellagic acidellagic acid concentrateAMES testL5178Y TK assay
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草莓粕是在榨草莓汁以釀製草莓酒時產生的副產物,先前已建立由草莓粕中回收鞣花酸 (ellagic acid, EA)的方法。本研究目的為簡化回收鞣花酸濃縮物之流程,並以安氏試驗法(AMES test)及L5178Y TK分析法來評估鞣花酸標準品 (Ellagic acid standard, EAS),及鞣花酸濃縮物(Ellagic acid concentrate, EAC)的安全性。在溶劑萃取鞣花酸方面,結果發現在甲醇、乙酸乙酯、丙酮和95%乙醇等四種溶劑中,EA萃取回收率以甲醇最佳約67%,其次為丙酮,約53%,再次是95%乙醇,約32%,而乙酸乙酯無法萃出EA。以甲醇及丙酮萃取2次後,再以乙酸乙酯處理萃取物以去除雜質,結果顯示EA幾乎不損失,且EA濃度度由原酸水解後草莓粕原料的0.4%提高至9.6%,乙酸乙酯用量也較原方法減少。在安全性評估方面,安氏試驗結果顯示不論添加S9與否,EAS與EAC對Salmonella typhimurium的五株菌(TA100, TA98, TA102, TA1535和TA97)都沒有顯著的致突變性(p<0.05)。EAS對L5178Y細胞之50%致死率為20 ppm,而EAC在最高濃度(10 ppm)時仍不具任何細胞毒性。EAS及EAC相較於溶劑對照組,突變頻率(mutation frequency)無顯著的差異(p<0.05)。
Strawberry pomaces are the byproducts of brewing strawberry wine. In our previous study, a ellagic acid (EA) recovery process from the strawberry pomaces had well been established. The aim of this study, we try to simplify the EA recovery process from strawberry pomaces and evaluate the safety of products, ellagic acid concentrate (EAC), and ellagic acid standard (EAS) by AMES test and L5178Y TK assay. The result of modified processes for recovering ellagic acid was indicated that a more efficient method has been reached by changing the extraction steps according to the properties of solvents. The ellagic acid content of final products (EAC) was 9.6% and ellagic acid was almost fully recovered from raw materials. The results of safety evaluation indicate that neither EAS nor EAC reveal any evidence of mutagenicity sn Salmonella typhimurium five test strains (TA98, TA100, TA102, TA1535 and TA97) either in the presence or absence of S9 activation. EAS showed cytotoxicity at 20 ppm and EAC cause no cytotoxicity at highest concentration (10 ppm). EAS and EAC had no significant mutagenicity compared with negative control of DMSO solvent (p<0.05).
目次………………………………….…………………………………………………..1
圖次……………………………………………………………………………………...3
表次……………………………………………………………………………………...4
中文摘要………………………………………………………………………………...5
英文摘要Abstract……………………………………………………………………….6
第一章、 研究動機…………………………………………………………………….7
第二章、 文獻整理…………………………………………………………………….9
壹、 自然界中的鞣花酸…………………………………………………………...9
貳、 鞣花酸之基本性質………………………………………………………….11
參、 鞣花酸之製備……………………………………………………………….16
肆、 鞣花酸之生理活性………………………………………………………….18
I. 抗氧化與抗癌活性…………………………………………………….18
II. 抗致突變性及抗腫瘤發生….…………………………………………20
III. 鞣花酸之美白功效…………………………………………………….21
IV. 鞣花酸之其他生理活性…………………………………………….....22
伍、 鞣花酸之生物可利用率及生物體內之代謝……………………………….23
陸、 鞣花酸之安全性…………………………………………………………….24
柒、 安全性評估方法…………………………………………………………….25
捌、 安氏試驗法 AMES test……………………………………………………..26
玖、 L5178Y TK分析法……………………………………………………….....28
第三章、 材料方法與結果...........................................................................................30
第一部份:鞣花酸濃縮物製程之改進.................................................................30
壹、 實驗架構…………………………………………………………………….30
貳、 實驗材料與器材…………………………………………………………….31
參、 實驗步驟…………………………………………………………………….32
肆、 實驗結果與討論…………………………………………………………….35
第二部份:鞣花酸標準品(EAS)與鞣花酸濃縮物(EAC)之安全性評估.............45
壹、 實驗架構………………………………………………………………….....45
貳、 安氏試驗法.....................................................................................................46
I. 實驗材料與器材….……………………………………………………46
II. 實驗步驟……………………………………………………………….52
III. 實驗結果與討論……………………………………………………….55
參、 L5178Y TK分析法.........................................................................................60
I. 實驗材料與器材……………………………………………………….60
II. 實驗步驟……………………………………………………………….61
III. 實驗結果與討論…….…………………………………………………65
第四章、 結論...............................................................................................................70
第五章、 參考文獻.......................................................................................................71
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