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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 郭錦樺(Ching-Hua Kuo) | |
| dc.contributor.author | Sung-Jeng Tsai | en |
| dc.contributor.author | 蔡松政 | zh_TW |
| dc.date.accessioned | 2021-06-17T00:30:37Z | - |
| dc.date.available | 2014-03-02 | |
| dc.date.copyright | 2012-03-02 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-02-13 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66330 | - |
| dc.description.abstract | 脂肪酸是體內參與能量平衡、訊息傳遞與維持細胞膜結構的重要物質。近幾年脂質代謝體的快速發展，配合化學劑量法分析大量樣品，發現血漿中許多脂質與疾病具相關性。本研究建立氣相層析質譜儀分析血漿中脂肪酸代謝體圖譜的方法，並比較不同的衍生化試劑，希望提供快速、穩固的方法在以應用於血漿中脂肪酸代謝體分析。
本研究以Folch萃取方法萃取血漿中脂質，以血漿萃取物、25種脂肪酸標準品衍生物與37種酯化脂肪酸標準品，對不同層析管柱與溫度梯度進行最佳化與探討，之後以優化條件比較Acetyl chloride、Hydrochloric acid、Boron trifluoride、Sulfuric acid、Sodium methoxide等五種衍生化試劑的衍生化效果，針對各種衍生化方法所提供之訊號強度和一日內與異日間之再現性進行討論。最佳化層析條件以SLB-IL82作為層析管柱，配合溫度梯度，可在25分鐘內建立血漿中脂肪酸之代謝體圖譜。相對其它衍生化方法，Acetyl chloride可以提供較好的訊號強度、再現性及較低的試劑價格。以最佳化的分析條件及Acetyl chloride衍生化方法分析心跳停止復甦急救之大鼠血漿，主成份分析結果顯示本研究所建立的脂肪酸代謝體圖譜可成功區分急救前、急救當下與急救後一小時的大鼠血漿。本實驗開發出一個快速且具有穩定再現性的分析方法，可以應用在未來分析脂質代謝體中脂肪酸的變化。 | zh_TW |
| dc.description.abstract | Fatty acids play important roles in the energy balance and signal transduction in biological systems. Some fatty acids are important constituents of cell membrane. Recent development in Lipidomics with the advance in chemometric techniques discovered many fatty acids are associated with numerous diseases. Gas chromatography-mass spectrometry (GC-MS) is the most commonly used analytical tool for fatty acid analysis. We compared five frequently used derivatization methods, and optimized GC-MS conditions. This study aimed to establish a fast, economic and robust GC-MS method for metabolic profiling of fatty acid in plasma.
We used the Folch method to extract fatty acids in plasma. Plasma extract, 25 endogenous fatty acid and 37 fatty acid methyl esters were used to optimize gas chromatographic conditions. By using a SLB-IL82 column with optimized temperature gradient, metabolic profiles of fatty acid in plasma can be constructed within 25 minutes. We used the optimal GC-MS conditions to compare the performance of five derivatization methods, including Acetyl chloride, Hydrochloric acid, Boron trifluoride, Sulfuric acid and Sodium methoxide. The performances of the derivatization method were compared in terms of signal intensity, method precision and reagent cost. Compared to other derivatization methods, Acetyl chloride provided the highest singal intensity, best method precision with minimal reagent cost, and it was selected as the derivatization reagent for metabolic profiling of fatty acid in plasma. The developed method was applied to investigate fatty acid changes in post-cardiac arrest myocardial dysfunction rats. The plasma samples were collected at three time points before and after ischemia reperfusion, and they analyzed by the developed method. The results indicated the metabolic profiles of three time points can be distinquished in the scores plot of principal component analysis. The fast and robust method appears to be applicable to detect changes in fatty acids in plasma samples. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T00:30:37Z (GMT). No. of bitstreams: 1 ntu-101-R98423015-1.pdf: 873571 bytes, checksum: c2d8b66e4685087a74c90bf99c58225b (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | 目 錄
中文摘要………………………………………………………………………………I 英文摘要……………………………………………………………………………...II 內文目錄…………………………………………………………………………….III 內文目錄 壹、序論 1.1 脂質代謝體學------------------------------------------------------------------------1 1.2 脂質分類及命名法------------------------------------------------------------------1 1.3 脂質分析方法------------------------------------------------------------------------3 貳、引言-------------------------------------------------------------------------------------------4 2.1 脂肪酸在疾病的重要性------------------------------------------------------------4 2.2脂肪酸分類與命名方法-------------------------------------------------------------4 2.3 現有分析脂肪酸的方法------------------------------------------------------------5 參、研究背景-------------------------------------------------------------------------------------7 3.1脂肪酸的重要性----------------------------------------------------------------------7 3.2 主要分析平台比較------------------------------------------------------------------7 3.3氣相層析分析方法開發重點--------------------------------------------------------8 肆、研究目的-----------------------------------------------------------------------------------12 伍、實驗部份----------------------------------------------------------------------------------13 5.1試劑------------------------------------------------------------------------------------13 5.2儀器------------------------------------------------------------------------------------13 5.3 GC-MS分析條件-------------------------------------------------------------------14 5.4標準品溶液製備---------------------------------------------------------------------14 5.5空白血漿製備------------------------------------------------------------------------15 5.6檢品溶液製備------------------------------------------------------------------------15 5.7衍生化方法---------------------------------------------------------------------------15 5.8 大鼠心跳停止並急救復甦的動物急救研究----------------------------------18 陸、結果 6.1 脂肪酸的萃取-----------------------------------------------------------------------18 6.2 GC-MS分析方法的最佳化-------------------------------------------------------20 6.3 衍生化方法的選擇-----------------------------------------------------------------23 6.4 分析方法的應用--------------------------------------------------------------------25 6.5 結論與未來展望--------------------------------------------------------------------26 柒、參考文獻-----------------------------------------------------------------------------------27 圖目錄 Fig 1. 不同管柱對於市售37酯化脂肪酸的層析圖-------------------------------------32 Fig 2. 三種升溫條件對(A)血漿樣品(B) 37個酯化脂肪酸與(C)25種脂肪酸標準品GC-MS圖譜------------------------------------------------------------------------------------33 Fig 3. 四種衍生化方法對26個內生性脂肪酸標準品訊號強度比較(n=3)---------36 Fig 4. 五種衍生化試劑對血漿樣品衍生化之訊號強度比較(n=3)-------------------37 Fig 5. 大鼠急救前、急救當下與急救後一小時之PCA scores plot -----------------38 Fig 6. 急救前、急救當下與急救後一小時之Heatmap -------------------------------39 Fig 7. 25個具有統計上顯著意義脂肪酸之箱形圖(boxplots)-------------------------41 表目錄 Table 1. 現有脂質資料庫-------------------------------------------------------------------42 Table 2. 脂肪酸編號與常用命名----------------------------------------------------------43 Table 3. 脂肪酸酸性與鹼性衍生試劑優缺比較----------------------------------------45 Table 4. 五種衍生化方法耗費時間與金錢比較----------------------------------------45 Table 5. 五種衍生化試劑優缺點比較----------------------------------------------------46 Table 6. 各種萃取方法的比較-------------------------------------------------------------48 Table 7. 二十六種脂肪酸標準品一日之內重覆性-------------------------------------49 Table 8. 脂肪酸定量離子、滯留時間與定性離子-------------------------------------50 Table 9. 五種衍生試劑衍生血漿樣品一日之內重覆性-------------------------------52 Table 10. 五種衍生化試劑衍生血漿樣品一日內相對標準偏差範圍的比較------53 Table 11. Acetyl chloride 與Boron trifluoride 衍生26種脂肪酸的標準品於異日間之再現性----------------------------------------------------------------------------------------54 Table 12. Acetyl chloride 與Boron trifluoride 兩衍生方法於血漿樣品一日內與異日間之再現性----------------------------------------------------------------------------------55 Table 13. Acetyl chloride 與Boron trifluoride 兩種衍生化方法衍生血漿樣品相對標準偏差範圍的比較-------------------------------------------------------------------------56 Table 14. 三個時間點中具有顯著差異的脂肪酸---------------------------------------57 | |
| dc.language.iso | zh-TW | |
| dc.title | 建立氣相層析質譜儀分析血漿中脂肪酸代謝體圖譜之方法 | zh_TW |
| dc.title | Development of a gas chromatography-mass spectrometry method for metabolic profiling of fatty acid in plasma | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇慧敏(HM Su),吳信隆(HL Wu) | |
| dc.subject.keyword | 脂肪酸,氣相層析質譜儀,SLB-IL82,脂質萃取,代謝體圖譜, | zh_TW |
| dc.subject.keyword | fatty acids,GC-MS,SLB-IL82,lipid extraction,metabolic profile, | en |
| dc.relation.page | 59 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-02-13 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 藥學研究所 | zh_TW |
| 顯示於系所單位： | 藥學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-101-1.pdf 目前未授權公開取用 | 853.1 kB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。