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研究生:林政葦
研究生(外文):Cheng-Wei Lin
論文名稱:衍生化氣相層析法對5-fluorouracil與其前驅藥tegafur之分析研究
論文名稱(外文):Determination of 5-fluorouracil and its prodrug tegafur by derivatization and gas chromatography
指導教授:柯黃盛柯黃盛引用關係
指導教授(外文):Hwang-Shang Kou
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:藥學研究所碩士班
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:90
中文關鍵詞:tegafur5-fluorouracil氣相層析衍生化
外文關鍵詞:derivatizationgas chromatography5-fluorouraciltegafur
相關次數:
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  • 收藏至我的研究室書目清單書目收藏:1
本研究利用衍生化反應配合氣相層析-電子捕獲型檢測法(gas chromatography-electron capture detection,GC-ECD),對於人體血漿中癌症化學治療劑5-fluorouracil (5-FU)與其前驅藥tegafur (TF;Ftorafur),建立一簡單且高感度之微量分析法。TF與5-FU以乙腈(acetonitrile)為反應溶媒,2,2’-dinitrobiphenyl為內部標準品, 18-crown-6 ether與碳酸鉀為催化劑,與酸可移除性衍生試劑2-(pentafluorophenoxy)ethyl 2-(piperidino)ethanesulfonate (PFPES)進行衍生反應,形成具有良好電子親和性之衍生物,所得之衍生物利用毛細管氣相層析儀來分離,以電子捕獲型檢測器 (ECD)進行偵測,而其偵測極限為:TF,1 �嵱;5-FU,0.1 �嵱 (S/N=3,注入體積1 �尳)。
影響TF與5-FU衍生化反應之重要因素,包括衍生試劑濃度、18-crown-6 ether濃度、鹼催化劑需要量、反應溫度及時間等,皆詳加探討。
本法應用於人體血漿中TF與5-FU之分析,僅使用少量血漿檢品 (200 μL),利用乙腈與乙酸乙酯四比六之比例萃取。可測定之線性範圍TF為5-250 �嵱 (1~50 μg /mL),5-FU為0.5-125 �嵱 (0.065~16.25 μg /mL),相關係數r皆大於0.995。其萃取相對回收率大於98.07 %,說明本法具有良好之定量可靠性,可應用於分析人體血漿上。
Analytical derivatization coupled with gas chromatography-electron capture detection (GC-ECD), a simple and sensitive method has been developed for the quantitative analysis of 5-fluorouracil (5-FU) and its prodrug tegafur (TF; Ftorafur), an chemotherapeutic antineoplastic agent, in human plasma. TF and 5-FU were derivatized with 2-(pentafluorophenoxy)ethyl 2-(piperidino)ethanesulfonate (PFPES) in a acetonitrile, using potassium carbonate and 18-crown-6 ether as reaction activators. The resulting derivatives of TF and 5-FU were analyzed by capillary gas chromatography with an electron-capture detector (GC-ECD). 2,2’-dinitrobiphenyl was used as an internal standard (I.S.), and the peak area ratios of TF and 5-FU derivatives to I.S. were used for quantitative analysis. The detection limit (signal to noise ratio = 3, injected volume 1 �尳) of TF and 5-FU was about 1 �嵱 and 0.1 �嵱.
Several parameters affecting the derivatization of TF and 5-FU such as the concentration of derivatizating agent, the concentration of 18-crown-6 ether, the amount of base, the reaction temperature and reaction time were discussed.
Application of the method to the analysis of TF and 5-FU in human plasma was studied. TF and 5-FU were extracted with acetonitrile and ethyl acetate (40:60, v/v) from 200 �尳 plasma. The linear range for the quantitation of TF and 5-FU in spiked plasma was 5-250 �嵱 �v1~50 μg/mL) and 0.5-125 �嵱�v0.065~16.25 μg/mL) with a correlation coefficient of 0.995. The relative recovery for TF and 5-FU in plasma was greater than 98.07 %. The method can be used for analysis of TF and 5-FU in human plasma.
中文摘要-------------------------------------------------------------------------- i
英文摘要-------------------------------------------------------------------------- ii
圖目錄----------------------------------------------------------------------------- vii
表目錄----------------------------------------------------------------------------- x

第一章、緒論---------------------------------------------------------------------- 1

第二章、實驗部分---------------------------------------------------------------- 8
一、試藥及材料------------------------------------------------------------- 8
二、儀器---------------------------------------------------------------------- 10
三、衍生試劑2-(pentafluorophenoxy)ethyl 2-(piperidino)ethanesulfonate (PFPES)之合成--------------
12
四、試藥溶液之配製------------------------------------------------------ 15
五、氣相層析法之分析條件-------------------------------------------- 19
六、GC-MS之分析條件-------------------------------------------------- 20
七、Tegafur與5-fluorouracil衍生物之合成-------------------------- 21
八、Tegafur與5-fluorouracil之基本分析探討------------------------- 23
I. 適當衍生反應條件之探討-------------------------------------- 23
1. 衍生試劑濃度之探討-------------------------------------- 24
2. 反應溫度及時間之探討----------------------------------- 25
3. 18-Crown-6 ether濃度之探討----------------------------- 25
4. 鹼催化劑種類及需要量之探討-------------------------- 26
II. 檢量線之探討---------------------------------------------------- 27
III. 衍生物之安定性探討------------------------------------------ 28
九、Tegafur與5-fluorouracil之應用分析探討----------------------- 29
I. 血漿樣品萃取之探討-------------------------------------------- 29
1. 血漿樣品之前處理----------------------------------------- 29
2. 血漿樣品之萃取及衍生步驟----------------------------- 29
3. 萃取溶媒比例之探討-------------------------------------- 31
4. 萃取溶媒體積之探討-------------------------------------- 32
II.血漿樣品之適當衍生反應條件探討-------------------------- 33
1. 血漿樣品中衍生試劑濃度之探討----------------------- 33
2. 血漿樣品中反應溫度及時間之探討-------------------- 34
3. 血漿樣品中18-crown-6 ether濃度之探討------------- 35
4. 血漿樣品中鹼催化劑需要量之探討-------------------- 36
III. Tegafur與5-fluorouracil檢量線之探討及其精密度、準
確度之探討-------------------------------------------------------
37
IV. 人體血漿中萃取回收率之探討------------------------------ 39
V. 人體血漿中衍生物之安定性探討---------------------------- 40
VI. 人體血漿中選擇性之探討------------------------------------ 41

第三章、結果與討論------------------------------------------------------------- 42

一、Tegafur與5-fluorouracil衍生物之質譜分析-------------------- 42
二、反應溶媒之選擇------------------------------------------------------- 44
三、內部標準品的選擇--------------------------------------------------- 45
四、基本分析最適當衍生反應條件之建立--------------------------- 47
I. 適當衍生反應條件之建立-------------------------------------- 47
1. 衍生試劑濃度之決定-------------------------------------- 47
2. 反應溫度與時間之決定----------------------------------- 48
3. 18-Crown-6 ether濃度之決定----------------------------- 49
4. 鹼催化劑種類及需要量之決定 ------------------------ 50
II. 檢量線之建立---------------------------------------------------- 53
1. 衍生物之安定性-------------------------------------------- 53
2. 檢量線之建立----------------------------------------------- 54
五、應用分析最適衍生反應條件之建立------------------------------- 56
I. 適當衍生反應條件之建立-------------------------------------- 56
1. 萃取溶媒比例之決定-------------------------------------- 56
2. 萃取溶媒體積之決定-------------------------------------- 57
3. 衍生試劑濃度之決定-------------------------------------- 58
4. 反應溫度與時間之決定----------------------------------- 59
5. 18-Crown-6 ether濃度之決定----------------------------- 60
6. 碳酸鉀需要量之決定----------------------------------- 61
II. 檢量線之建立及其精密度準確度之測定------------------- 64
1. 衍生物之安定性-------------------------------------------- 64
2. 檢量線之建立----------------------------------------------- 65
3. 精密度及準確度之測定----------------------------------- 66
III. 人體血漿中tegafur與5-fluorouravil之萃取回收率------ 67
IV. 選擇性之試驗法------------------------------------------------ 68

第四章、結論---------------------------------------------------------------------- 69

第五章、參考文獻---------------------------------------------------------------- 70
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