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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝淑貞(Shu-Chen Hsieh) | |
dc.contributor.author | Yu-Hsiu Chang | en |
dc.contributor.author | 張羽秀 | zh_TW |
dc.date.accessioned | 2021-06-16T05:23:38Z | - |
dc.date.available | 2019-08-17 | |
dc.date.copyright | 2014-08-17 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-15 | |
dc.identifier.citation | 林孟儒 (2013) 中國橄欖甲醇萃取-乙酸乙酯區分層之促葡萄糖攝取功效探討 臺灣大學食品科技研究所碩士論文
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56327 | - |
dc.description.abstract | 中國橄欖(Canarium album)為橄欖科、橄欖屬。在中醫觀點中,因其具有抗菌、抗病毒、抗發炎與解毒之效果而被用以治療感染、痢疾及解蛇毒。本實驗室先前的研究發現中國橄欖甲醇萃取物正丁醇區分層((Methanol-butanol olive extract(MBO))在小鼠大腸癌細胞(CT26)中具有顯著抑制癌細胞增生之效果,因此將進一步利用人體細胞平台的檢測結果,分析主要抗癌細胞增生活性的成分,最後將討論其抗癌細胞增生的機制。先前文獻指出,許多天然物會影響粒線體活性,而影響依賴粒線體酵素活性的細胞存活率測試方法之吸光值(如:MTT assay);因此透過與IN Cell Analyzer 2200確認不同細胞存活率方法學的準確性,該儀器透過螢光染劑(DAPI)將細胞核中染色質染色後,由儀器計數得到準確細胞數。根據實驗結果,選擇MTT assay做為抗癌細胞增生活性測試的方法。樣品的部分,本實驗選用新竹縣寶山鄉種植之中國橄欖,收集水萃後之殘渣以甲醇萃取後減壓濃縮,再利用液-液萃取(liquid-liquid partition)的方式,以正己烷、乙酸乙酯及正丁醇等與水不互溶之有機溶劑,將此中國橄欖甲醇萃取物進行極性切割。再將中國橄欖甲醇-正丁醇區分層(MBO)在不同人類細胞平台進行活性測試,發現其在人類之多種大腸癌 (HCT116、HCT116 p53-/-、CaCO2)、舌癌(Ca922)、肺癌(A549)、血癌(HL-60)…等細胞中亦具有良好之抗癌細胞增生的效果。進一步將中國橄欖甲醇萃取物正丁醇區分層(MBO)利用酸、中、鹼三種不同pH值的緩衝溶液處理,藉此改變物質溶解度後,再以正丁醇反萃出不同酸鹼值的物質進行抗癌細胞增生活性篩選;發現中國橄欖甲醇-正丁醇區分層(MBO)經酸處理後具有最佳抗癌細胞增生效果。因此將此酸處理後的物質利用HP20疏水樹脂逆相管柱層析,以五種不同濃度的甲醇依序進行流洗,發現E4之次區分層保有最好的活性。而進一步以HPLC檢測,發現其含有高達72 %的鞣花酸。故推測鞣花酸為中國橄欖甲醇萃取物正丁醇區分層抗癌細胞增生之主要有效成分之一,至於其他波峰為何則需要進一步的鑑定分析。 | zh_TW |
dc.description.abstract | Chinese olive (Canarium album.), which belongs to family of Burseraceae and genus of Canarium, is a traditional Chinese medicine with antibiotic, anti-virus, anti-inflammatory, as well as detoxification effects, and is used for curing infection, malaria, venom poinsoning. Former results in our lab indicated that the methanol-butanol olive extraction (MBO) significantly repressed the viability of murine colon cancer cells (CT26); however, more investigations for verifying the bioactive compounds in human cells model are needed. In the present study, Chinese olives debris was collected after water extraction, and extracted again with methanol. The methanol extract of Chinese olives was then fractioned by liquid-liquid partition with hexane, ethyl acetate, and butanol. The MBO was collected for further anti-cancer analysis. On the other hand, previous studies in our lab or other lab shows that some phytochemicals increase the activity of enzymes in mitochondria, making mitochondria-dependent viability assays, such as MTT become imprecise. To ascertain the accuracy of methodology, comparison of WST-1 and MTT with the control of cells counting by IN Cell Analyzer 2200 were conducted, and MTT assay was chosen to be the optimal way to determine the cancer cell viability. Results showed that MBO possessed anti-proliferative capacity against several cancer cells including colon cancer cells of HCT116 and HCT116 p53-/-, tongue cancer cell (Ca922), lung cancer cells (A549), and blood cancer cells (HL-60). Besides, MBO didn’t repress the proliferation of normal cell, including Int407 and CaCO2. After processing MBO with acidic buffer solution, MBO-2B fraction possessed enhancing the anti-tumor ability. Furthermore, fractionation the MBO-2B by HP20 resin, obtaining E4 sub-fraction with enriched anti-proliferative effect. By HPLC, compound of Ellagic acid accounted for 72 % of this elution. Hence, Ellagic acid was believed as the functional compound for the antitumor property in Chinese olives. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T05:23:38Z (GMT). No. of bitstreams: 1 ntu-103-R01641037-1.pdf: 3271472 bytes, checksum: 32800e65c1cb3892602fd225f6cfb6e2 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 目錄
口委審定書………………….………………………………………………………….I 謝誌………………….………………………………………………………………….II 摘要…………………………………………………………………………………….IV Abstract…………………………………..…………………………………………….V 目錄…………………………………………………………..……………….……….VI 圖目錄……………………………………………………………………..………… VIII 表目錄……………………………………………………………………..………… IX 縮寫表……………………………………………………………………..………… X 第一章、前言 1 第二章、文獻回顧 2 第一節 、中國橄欖(Chinese olive) 2 一、 中國橄欖背景介紹 2 二、 中國橄欖的成分 2 三、 中國橄欖的生理功能 3 第二節 、癌症 5 一、 癌細胞的特色 5 二、 癌症的發展 7 三、 癌症與化學預防 8 第三節 、單寧 9 一、單寧(Tannin)的介紹 9 二、 單寧的萃取方法 16 三、 常見的單寧定量分析方法 17 四、 單寧的生物可利用性 18 五、 單寧發揮生理活性的作用方式 19 六、 單寧的生理功能 20 第四節 、細胞存活率的方法學 23 一、 常見細胞存活率測試方法學整理 23 二、 依賴細胞酵素代謝活性的WST-1 assay及MTT assay 24 三、 植化素對依賴粒腺體酵素活性測定細胞存活率方法學(MTT)的影響 27 第三章 實驗動機與目的 28 一、實驗動機 28 二、實驗目的 28 第四章 材料與方法 30 第一節 、篩選中國橄欖甲醇萃取物正丁醇區分層中具抗癌細胞增生的活性成分 30 第二節 、細胞存活率平台挑選 32 第三節 、中國橄欖萃取及化學分析 34 第五章 結果與討論 41 第一節 、中國橄欖甲醇萃取物正丁醇區分層之品質管控(Quality control) 41 第二節 、細胞存活率(cell viability)方法學比較 46 第三節 、中國橄欖甲醇萃取物正丁醇區分層(MBO)之抗癌細胞增生活性 50 第四節 、中國橄欖甲醇萃取物正丁醇區分層(MBO)之抗癌細胞增生活性成分之化學分析 54 第六章 結論 70 第七章 參考文獻 71 圖目錄 圖 一、縮合型單寧基本結構 12 圖 二、縮合型單寧的分類:依單體間鍵結型式 12 圖 三、縮合型單寧的分類:依氫氧基位置 12 圖 四、水解型單寧的生合成機制 14 圖 五、沒食子酸(Gallic acid) 15 圖 六、鞣花酸(Ellagic acid) 15 圖 七、MTT、WST-1之結構與其在細胞代謝的位置示意圖 26 圖 八、MBO之HPLC圖譜分析 43 圖 九、沒食子酸(Gallic acid)之HPLC標準曲線 45 圖 十、鞣花酸 ( Ellagic acid ) 之HPLC標準曲線 45 圖 十一、利用中國橄欖甲醇萃取物正丁醇區分層測試各種細胞存活率方法學的精確度分析 49 圖 十二、中國橄欖甲醇萃取物正丁醇區分層(MBO)抗不同癌細胞株增生效果 52 圖 十三、中國橄欖甲醇萃取物正丁醇區分層(MBO)抗正常細胞增生效果 53 圖 十四、中國橄欖甲醇萃取物正丁醇區分層(MBO)調酸中鹼處理後對HCT116人類大腸癌細胞株抗增生效果評估 57 圖 十五、中國橄欖甲醇-正丁醇區分層(MBO)調酸中鹼處理後對HCT116人類大腸癌細胞株抗增生效果評估(修正版) 59 圖 十六、酸處理之中國橄欖甲醇萃取物正丁醇區分層(MBO-2B)經HP20管柱層析之次區分層對HCT116人類大腸癌細胞株抗增生能力 64 圖 十七、酸處理MBO之HPLC圖譜 65 圖 十八、E2、E3、E4之HPLC圖譜 66 圖 十九、E2、E3、E4以PVPP處理之HPLC圖譜 67 圖 二十、E2、E3、E4以酸水解處理之HPLC圖譜 68 表目錄 表 一、中國橄欖核三大營養素含量(g / 100g 乾重) 3 表 二、單寧的分類及其單體結構 10 表 三、中國橄欖甲醇萃取物及其液-液萃取區分層萃取率 42 表 四、中國橄欖甲醇萃取物正丁醇區分層(MBO)之指標成分定量 44 表 五、中國橄欖甲醇萃取物正丁醇區分層(MBO)對不同癌細胞株之半數致死濃度(IC50) 51 表 六、中國橄欖甲醇-正丁醇區分層(MBO)調酸、中、鹼處理之半數致死濃度(IC50)及產率 56 表 七、中國橄欖甲醇萃取物正丁醇區分層(MBO)調酸中鹼處理之半數致死濃度IC50及產率(修正版) 58 表 八、酸處理之中國橄欖甲醇萃取物正丁醇區分層(MBO-2B)經HP20管柱層析之次區分層之半數致死濃度(IC50)及萃取率 63 表 九、鞣花酸在不同流洗條件下的產率、純度及半數致死濃度(IC50) 69 | |
dc.language.iso | zh-TW | |
dc.title | 以活性導向方式分離中國橄欖果實內具抑制癌細胞增生之成分 | zh_TW |
dc.title | Bioassay-guided Isolation of Active Components from the Fruit of Canarium album toward Inhibition of Cancer Cell Growth | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 蘇南維(Nan-Wei Su) | |
dc.contributor.oralexamcommittee | 潘敏雄(Min-Hsiung Pan),郭靜娟(Ching-Chuan Kuo) | |
dc.subject.keyword | 中國橄欖,抑制癌細胞增生,鞣花酸,中國橄欖甲醇萃取物正丁醇區分層,液液萃取, | zh_TW |
dc.subject.keyword | Chinese olives,anti-proliferative,ellagic acid,Methanol-butanol olive extract, | en |
dc.relation.page | 77 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-08-15 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 食品科技研究所 | zh_TW |
顯示於系所單位: | 食品科技研究所 |
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