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研究生:謝俊宇
研究生(外文):Chun-Yu Hsieh
論文名稱:衍生化氣相層析對甲苯之代謝物o-Cresol及Hippuricacid之分析及應用
論文名稱(外文):Analysis and Application of Toluene Metabolites:o-Cresol and Hippuric Acid
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
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:2002
畢業學年度:90
語文別:中文
論文頁數:67
中文關鍵詞:甲苯
外文關鍵詞:tolueneo-cresolhippuricacid
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甲苯(Toluene)目前在工業上是一個廣泛被使用的有機溶媒。目前用來評估甲苯暴露量的最好方法,就是監測人體尿液中甲苯代謝產物hippuric acid (HA)及o-cresol的濃度,因為此兩者與甲苯的暴露量有較明顯的相對關係。
本研究以親電子性衍生試劑2-(pentafluorophenoxy)ethyl- 2-(piperidino)ethanesulfonate (PFPES)及碳酸鉀與18-crown-6 ether為催化劑對甲苯代謝之生物指標物o-cresol及HA建立一種簡單及高感度分析法,此o-cresol及HA衍生物以氣相層析配合電子捕獲型偵測器(GC-ECD)進行微量分析。定量範圍為1.0~50 µM之o-cresol及10~200 µM之HA,最低檢測極限為o-cresol 0.1 µM(S/N = 3)(每1µL注入量含0.1 pmol)及HA 1.0 µM(S/N = 3)(每1µL注入量含1.0 pmol)。應用此方法分析尿液中之o-cresol及HA,目前仍在進行中。
Toluene is widely used as an organic solvent in industry. The best estimation for assessment of exposure to toluene is the quantitative determination of the metabolites, hippuric acid (HA) and o-cresol, excreted in urine, which show good correlation between the level of exposure and amount of metabolites excreted.
A highly sensitive method is described for the determination of o-cresol and HA, a biomarker of toluene exposure. The method is based on the derivatization of HA and o-cresol with an electrophoric reagent, 2-(pentafluorophenoxy)ethyl 2-(piperidino)ethanesulfonate, using potassium carbonate and 18-crown-6 ether as reaction activators. The resulting pentafluorophenoxy derivative of HA and o-cresol was analyzed by capillary GC with an electron-capture detector (ECD). The quantitation range of o-cresol and HA are 1.0~50 µM and 10~200 µM, respectively. The detection limit of o-cresol and HA are 0.1 µM and 1.0 µM, respectively (S/N = 3). Application of the method to the analysis of o-cresol and HA in urine is be investigated.
中文摘要…………………………………………………… iv
英文摘要…………………………………………………… v
壹、 緒論……………………………………………… 1
貳、 實驗部分………………………………………… 6
一、 壹、 試藥與材料……………………………………… 6
二、 貳、 試藥溶液之配製…………………………………A. o-Cresol試藥溶液之配製……………………B. Hippuric acid 試藥溶液之配製………….…..C. 硫酸溶液之配製……………………………..D. 尿液檢品之配製………………………….… 88101313
三、 肆、 儀器……………………………………………… 14
四、 氣相層析條件…………………………………… 15
五、 基本分析法之探討………………... ………….... 16
A. o-Cresol適當衍生條件之探討………………...... 16
A-1) 內部標準品之選擇………………………... 16
A-2) 反應系中有機溶媒之選擇………………... 17
A-3) 反應系中衍生試劑濃度之探討………….. 17
A-4) 反應系中鹼催化劑需要量之探討………... 18
A-5) 反應系中18-crown-6 ether濃度之探討…. 19
A-6) 反應系中反應溫度與時間之探討……….. 19
A-7) o-Cresol檢量線之製定法…………………. 19
A-8) o-Cresol衍生物安定性之探討……………. 20
A-9) o-Cresol衍生物之結構確認………………. 20
B. Hippuric acid適當衍生條件之探討……………. 21
B-1) 內部標準品之選擇………………………... 22
B-2) 反應系中有機溶媒之選擇………………... 23
B-3) 反應系中衍生試劑濃度之探討………….. 23
B-4) 反應系中鹼催化劑需要量之探討………... 23
B-5) 反應系中18-crown-6 ether濃度之探討…. 24
B-6) 反應系中反應溫度與時間之探討……….. 24
B-7) Hippuric acid檢量線之製定法…………….. 25
B-8) Hippuric acid衍生物安定性之探討………. 25
B-9) Hippuric acid衍生物之結構確認…………. 26
六、 應用分析法之探討…………………………….... 27
A) 選擇性之探討……………………………… 27
B) 尿液檢品之前處理探討…………………….. 1. o-Cresol…………………………………… 2. Hippuric acid……………………………… 272728
參、 結果與討論……………………………………… 30
一、 基本分析法之探討……………………………… 30
A. o-Cresol適當衍生條件之探討………………..... 30
A-1) 內部標準品的選擇………………………... 30
A-2) 反應系中有機溶媒之選擇………………... 32
A-3) 反應系中衍生試劑濃度之決定………….. 33
A-4) 反應系中鹼催化劑之決定………………... 34
A-5) 反應系中18-crown-6 ether濃度之決定…. 35
A-6) 反應系中反應溫度與時間之決定……….. 36
A-7) o-Cresol衍生物之GC-ECD層析圖………. 38
A-8) o-Cresol檢量線之製定法………...……….. 40
A-9) o-Cresol衍生物安定性之探討……………. 40
A-10) o-Cresol衍生物之結構確認……………… 41
B. Hippuric acid適當衍生條件之探討…………….. 46
B-1) 內部標準品的選擇………………………... 46
B-2) 反應系中有機溶媒之選擇………………... 48
B-3) 反應系中衍生試劑濃度之決定………….. 48
B-4) 反應系中鹼催化劑需要量之決定………... 49
B-5) 反應系中18-crown-6 ether濃度之決定…. 50
B-6) 反應系中反應溫度與時間之決定……….. 50
B-7) Hippuric acid衍生物之GC-ECD層析圖… 53
B-8) Hippuric acid安定性之探討……………….. 55
B-9) Hippuric acid檢量線之製定法…………….. 55
B-10) Hippuric acid衍生物及結構確認………… 56
二、 應用分析法之探討……………………………… 60
A) 選擇性之探討………………………………. 60
B) 尿液檢品之前處理探討…………………….. 1. o-Cresol…………………………………… 2. Hippuric acid……………………………… 636364
肆、 結論……………………………………………… 66
伍、 參考文獻………………………………………… 67
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