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研究生:徐偉勛
研究生(外文):Wei-Hsun Hsu
論文名稱:以頂空固相微萃取法配合離子阱氣相層析質譜儀偵測尿液中的克倫特羅
論文名稱(外文):Determination of Clenbuterol in Urine by Head-space Solid Phase Microextraction and Gas Chromatograph-Ion Trap Mass Spectrometer
指導教授:黃賢達黃賢達引用關係
指導教授(外文):Shang-Da Huang
學位類別:碩士
校院名稱:國立清華大學
系所名稱:化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:71
中文關鍵詞:氣相層析質譜儀衍生化克倫特羅
外文關鍵詞:GC-MSDerivativesClenbuterol
相關次數:
  • 被引用被引用:4
  • 點閱點閱:176
  • 評分評分:
  • 下載下載:35
  • 收藏至我的研究室書目清單書目收藏:0
本論文主要是利用頂空固相微萃取法配合氣相離子阱層析質譜儀(HSSPME-GC/ITMS)並使用衍生化法來分析人體尿液中的一種合成代謝劑:克倫特羅(Clenbuterol)。

固相微萃取法是近年來所使用的一種前處理技術,它可以一次做好採樣、萃取、濃縮以及樣品導入的步驟,讓我們在做前處理時能更方便以及快速。而本論文的目的就是在利用這種技術再配合氣相層析質譜儀分析人體尿液中克倫特羅的含量。這篇所探討的重心在各種影響萃取效率的變因做探討,包括;萃取所用的纖維種類、待測物的萃取溫度、萃取時間與冷卻系統、脫附溫度和脫附時間、溶液的攪拌速率與酸鹼度、樣品中的離子強度等八項。以求得最佳的實驗條件,再用這條件(纖維:PA、待測物萃取時間、溫度:90分鐘、90℃、待測物上端溫度5℃、衍生劑萃取時間、溫度:5分鐘、70℃、樣品攪拌速率:300rpm、樣品酸鹼值:12、離子強度:添加30%的氯化鈉、脫附時間、溫度:3分鐘、260℃)來應用在尿液中。

而本論文最後是使用PA纖維來做偵測。由實驗結果可以發現其偵測極限可達到ng/mL以下的層級,且在1~1000 ng/mL的濃度範圍下能擁有相當高的線性相關係數(0.9990)。而且精密度可以在4%以下。可證實此方法用於偵測克倫特羅的可行性。
In this thesis, we develop a method of using headspace solid-phase microextraction (HS-SPME) combined with gas chromatograph-ion trap mass spectrometer (GC-ITMS) to detect a β-agonist : clenbuterol.

We discuss and optimize eight parameters of extraction and desertion procedure, including types of fibers, extraction temperature, extraction time, desorption temperature, desorption time, stir rate, ionic strength, pH of sample and cooling system.

The method show good linearity over the concentration range 1–1000 ng/mL and gives detection limits below ng/mL level. Precision is also good (3.89%). Finally, the method is successfully applied to analyze clenbuterol in urine.
一、緒論..............................................1
1-1前言...............................................1
1-2合成代謝劑.........................................3
1-3固相微萃取法.......................................4
1-3-1固相微萃取裝置演進...............................4
1-3-2固相微萃取的實驗步驟.............................4
1-3-3固相微萃取的原理.................................4
1-3-4操作模式.........................................6
1-3-5固相微萃取的應用.................................7
1-4衍生化法...........................................9
1-5 Clenbuterol的檢測方法............................10
1-5-1酵素免疫連結吸附分析法..........................10
1-5-2高效能液相層析法................................10
1-5-3氣相層析法......................................11
1-6研究目的..........................................13
二、實驗部分.........................................14
2-1實驗藥品..........................................14
2-2實驗裝置..........................................14
2-3實驗儀器..........................................15
2-4標準溶液..........................................16
2-5真實樣品..........................................17
2-6實驗步驟..........................................17
三、結果與討論.......................................20
3-1各種變因之探討....................................20
3-1-1衍生反應........................................20
3-1-2纖維種類........................................20
3-1-3萃取時間........................................22
3-1-4攪拌速率........................................23
3-1-5脫附時間........................................24
3-1-6脫附溫度........................................25
3-1-7酸鹼值..........................................25
3-1-8離子強度........................................26
3-1-9萃取溫度與冷卻系統..............................28
3-1-10最佳化條件.....................................30
3-2線性、精密度及方法偵測極限........................31
3-3真實樣品的測試....................................32
四、結論.............................................33
五、參考文獻.........................................34

表一、Clenbuterol的性質..............................42
表二、HMDS的性質.....................................43
表三、固相微萃取纖維之調態與操作溫度條件.............44
表四、商業化固相微萃取纖維的比較.....................45
表五、吸附型與吸收型靜相纖維的比較...................46
表六、最佳化條件選擇結果.............................47
表七、線性係數、精密度及方法偵測極限.................48
表八、實驗結果與相關文獻的比較.......................49

圖一、Clenbuterol、HMDS、Clenbuterol-TMS的結購.......50
圖二、Pawliszyn實驗室最早設計的固相微萃取裝置圖......51
圖三、商業化的手動固相微萃取裝置圖...................52
圖四、固相微萃取裝置的操作流程圖.....................53
圖五、三種不同的固相微萃取操作模式...................54
圖六、直接注射1μl ,1 ppm的已衍生完的Clenbuterol-TMS溶液所得到的層析圖...........................................55
圖七、Clenbuterol-TMS 的質譜圖.......................56
圖八、Clenbuterol-TMS經EI撞擊後可能分裂的六離子......57
圖九、固相微萃取裝置架設圖...........................58
圖十、直接注射1μl ,2 ppm的標準Clenbuterol溶液所得到的層析圖...................................................59
圖十一、各種纖維之性質差異...........................60
圖十二、纖維種類與萃取效率之關係圖...................61
圖十三、萃取時間與萃取效率之關係圖...................62
圖十四、攪拌速率與萃取效率之關係圖...................63
圖十五、脫附時間與萃取效率之關係圖...................64
圖十六、脫附溫度與萃取效率之關係圖...................65
圖十七、酸鹼值與萃取效率之關係圖.....................66
圖十八、鹽類濃度與萃取效率之關係圖...................67
圖十九、萃取溫度與萃取效率之關係圖...................68
圖二十、冷卻系統的存在與否與萃取效率之關係圖.........69
圖二十一、以最佳化條件萃取真實尿液中所得到的層析圖...70
圖二十二、Clenbuterol-TMS 在標準樣品與真實人體尿液中質譜圖的比較.................................................71
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