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研究生：	陳瑞慈 chen, Ray-Tzu
論文名稱：	非洲白蔘生物活性物質之純化與鑑定 Purification and Characterization of Bioactive Components from Mondia whitei
指導教授：	黃福永 Huang, Fu-Yung
學位類別：	碩士 Master
系所名稱：	理學院 - 化學系 Department of Chemistry
論文出版年：	2009
畢業學年度：	97
語文別：	中文
論文頁數：	82
中文關鍵詞：	美白、抗氧化、抗菌、非洲白蔘
外文關鍵詞：	Mondia Whitei, antibacterial tests, tyrosine inhibitor activity, antioxidant activity
相關次數：	點閱：159 下載：0
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非洲白蔘萃取物目前被用來測試生理活性的都是水萃物，為了瞭解生理活性的有效成份及其作用機制，成份之分離與鑑定是必要的。發現二氧化碳超臨界萃取可萃出具有辛辣味的成份。
將非洲白蔘的超臨界流體萃取液進行微生物試驗，利用Disc diffusion method測試，發現對黴菌(mold)及三種酵母菌(yeast)，Aspergillus niger 16404、Malassezia furfur 12078與Candida albicans 10231，有很好的抑制效果。對Malassezia furfur 12078進行活性測試各個分離程序，予以追蹤活性存在的部分，以活性為指標的分離程序(activity guided fractionation)，分離非洲白蔘活性成分。分離出純物質，純化之白色針狀固體經鑑定後，結構為2-Hydroxy-4-methoxybenzaldehyde和4-Methoxybenzaldehyde，對Malassezia furfur 12078抗菌效果良好。
對DPPH自由基清除能力測定結果以超臨界流體萃取物最好，經過半製備型高效能層析管柱進行分離，纯化出一白色結晶物，鑑定結構為4-Hydroxy-2-methoxybenzaldehyde，對DPPH自由基清除能力為86.09 %(1 mg/ml)，IC50為0.406 mg/ml。由不同溶劑萃取之非洲白蔘萃取物經酪胺酸酶抑制率力測定結果，證實其二氯甲烷萃取物最具活性。

So far, most bioactivity assay for Mondia Whitei were the aqueous and methanol extracts. In this study, CO2 super fluid critical (SFC) extraction has been employed and found that the extract contains spicy flavor. This extract was purified and identified for further bioassay. Three compounds, 2-Hydroxy-4-methoxybenzaldehyde, 4-Methoxybenzaldehyde, and 4-Hydro- xy-2-methoxybenzaldehyde, were identified.
The antibacterial tests for compounds, 2-Hydroxy-4-methoxybenza- ldehyde and 4-Methoxybenzaldehyde, against mold and three types yeast, Aspergillus niger 16404, Malassezia furfur 12078 and Candida albicans 10231, showed good activity, especially Malassezia furfur 12078.
Antioxidant activity against DPPH showed that crude CO2 SFC extract has better activity than crude queous or methanol extract. Among them, 4-Hydroxy-2-methoxybenzaldehyde showed 86.09 % (1 mg/ml) activity and the dose for IC50 is 0.406 mg/ml. With the concentration of 50 mg/ml for six extracts by H2O, Methanol, Ethyl acetate, CH2Cl2, n-Hexane, and CO2 SFC, for Tyrosine inhibitor activity, it was found that CH2Cl2 extract shows best result.

中文摘要………………………………………………………...I
Abstract……………………………………………………....II
致謝…………….......................…………………III
總目錄…………………………....................……….V
表目錄…………………………………………………..……VIII
圖目錄…………………………………………………...…….IX
第一章 緒論………………………………………………………1
1 非洲白蔘(Mondia Whitei)簡介………….……….………1
2 非洲白蔘(Mondia Whitei)文獻回顧..………..........5
2.1 非洲白蔘(Mondia Whitei)化學成分.….…......5
2.2 非洲白蔘(Mondia Whitei)藥理測.……….......6
2.3 非洲白蔘(Mondia Whitei)安全性評…..........8
3 非洲白蔘萃取物成份之分離概述.……..…………....…9
3.1 超臨界流體(SFC)的簡介…………………….....12
3.2 採用超臨界流體(SFC)優點、原理及技術的應用.14
3.3 超臨界萃取(SFC)的技術原理.................16
4 抗菌活性……………………………...……….........18
4.1 菌種簡介………………..…………..………....18
5 抗氧化能力…….................…………………….24
6 美白……………..……………………................28
6.1 黑色素形成和酪胺酸?活性作用的關係..….....28
6.2 酪胺酸?.……..............................28
6.3 還原劑對色素生成的影響....................30
第二章 實驗方法………………………….................31
1 實驗用試劑、儀器設備…….…………….…….……….31
2 藥材萃取…………………………………………………..33
3 抗菌測試…………………………...…………….……..35
3.1菌株之活化………………………….......………35
3.2抗菌測試………………………………...........35
3.3正相矽膠柱層析分離抗菌物質……….……..……36
4 光譜之測定………………………………………..……..37
4.1 NMR光譜之測定……………………………….…..37
4.2 IR光譜之測定:紅外線光譜 (Infrared Spectra)…..37
5 抗氧化功能測試…………………………........……..38
5.1 清除DPPH自由基能力之測定………….........38
5.2  HPLC層析分離抗氧化物質……………….…....39
6 對酪胺酸?活性抑制之測定……………...………..…..40
第三章 結果與討論…………………………………….......41
1 不同溶劑黃芩萃取物…………………..……..........41
2超臨界流體萃取最佳化條件…………………...........42
3 有機溶劑萃取之HPLC成份分析之結果與討論…...…...43
4 抗菌………………………………………………….…….46
4.1 抗菌活性測定……………...…………………...46
4.2 活性成分之分離與純化…………………………..50
4.3 抗菌活性成分(Mal-U)結構之鑑定………..…...51
4.4 抗菌活性成分(Mal-D)結構之鑑定……………….55
5 抗氧化……………………………………………………..59
5.1 抗氧化活性測定…………………………………..59
5.2 活性成分之分離與純化……………….………...62
5.3 抗氧化活性物質結構鑑定…………….……...…67
6  美白………………………...…………………………..71
6.1美白活性測定……….……………………………..71
第四章 結論…………………………………………………...74
參考文獻……………………………….………………….....76
                                    

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