臺灣博碩士論文加值系統

本研究利用衍生化高效能液相層析法，對苯之暴露指標物trans, trans-muconic acid建立簡單靈敏之分析方法。本法利用2-(2-naphthoxy)ethyl 2-(piperidino)ethanesulfonate (NOEPES)為衍生試劑，以氰甲烷為反應溶媒，於碳酸鉀及18-crown-6 ether之催化下，將trans, trans-muconic acid衍生爲naphthoxyethyl衍生物，以2-isobutoxynaphthalene爲內部標準，利用逆相C-18層析配合85﹪（v/v）甲醇水溶液為移動相；以紫外光（226 nm）偵測。所得結果顯示本法靈敏性高trans, trans-muconic acid之定量範圍爲0.3 ~ 20μM，偵測極限0.03μM (S/N = 3；20μL)。
本研究對影響trans, trans-muconic acid衍生化之重要因素，包括試劑需要量、反應溶媒、鹼性催化劑、反應時間及溫度等，皆詳加探討。
本法將應用於尿液中微量trans, trans-muconic acid之分析，目前正進行尿液中待測物之萃取及衍生化HPLC-UV分析探討。

Analytical derivatization coupled with high-performance liquid chromatography is used for the analysis of trans, trans-muconic acid, a biomarker of benzene exposure. The method is based on the derivatization of trans, trans-muconic acid with 2-(2-naphthoxy) ethyl 2-(piperidino) ethanesulfonate (NOEPES) in acetonitrile, using potassium carbonate and 18-crown-6 ether as reaction activators. The resulting naphthoxyethyl derivative of trans, trans-muconic acid was separated on a C-18 column, using a mixed solvent of methanol : water (85：15, v/v) as the mobile phase. The separated trans, trans-muconic acid derivative was monitored with UV at 226 nm. 2-Isobutoxynaphthalene was used as an internal standard (I.S.), and the peak-area ratios of trans, trans-muconic acid derivative to I.S. were used for quantitative analysis.
The linear range of the method for the determination of trans, trans-muconic acid was over 0.3 ~ 20μM. The detection limit (signal to noise ratio = 3, injected amount 20μL) of trans, trans-muconic acid was about 0.03μM. Several parameters affecting the derivatization of trans, trans-muconic acid were discussed including reaction solvent, amount of derivatization reagent, reaction temperature, reaction time, and base catalyst. Application of the method to the analysis of trans, trans-muconic acid in urine is being investigated.

圖目錄------------------------------------------------ ⅲ
表目錄------------------------------------------------ ⅴ
中文摘要---------------------------------------------- ⅵ
英文摘要---------------------------------------------- ⅶ
第一章、緒論------------------------------------------ 1
第二章、實驗部分-------------------------------------- 8
一、試藥及材料------------------------------------ 8
二、儀器------------------------------------------ 9
三、衍生試劑之合成-------------------------------- 10
四、試藥溶液之配製-------------------------------- 12
五、高效能液相層析之分析條件---------------------- 15
六、trans, trans-muconic acid衍生物之合成與精製--- 16
七、trans, trans-muconic acid衍生反應條件之探討--- 18
1、衍生試劑需要量----------------------------- 19
2、鹼性催化劑篩選----------------------------- 20
3、碳酸鉀需要量------------------------------- 21
4、18-crown-6 ether需要量--------------------- 22
5、反應溫度及時間----------------------------- 23
八、分析法之選擇性-------------------------------- 24
九、衍生物安定性---------------------------------- 25
十、應用分析-------------------------------------- 26
(一)、尿液中trans, trans-muconic acid之萃取--- 26
(二)、萃取所得trans, trans-muconic acid之衍
生HPLC分析------------------------------ 27
第三章、結果與討論------------------------------------ 28
一、trans, trans-muconic acid衍生物之質譜分析----- 28
二、偵測波長的決定-------------------------------- 29
三、反應溶媒之選取-------------------------------- 30
四、水層與有機層的分離---------------------------- 30
五、trans, trans-muconic acid衍生物之HPLC紫外光
層析圖與過量衍生試劑之酸洗移除性-------------- 32
六、內部標準的選擇-------------------------------- 34
七、最適衍生反應條件之建立------------------------ 35
1、衍生試劑需要量----------------------------- 35
2、鹼性催化劑篩選----------------------------- 37
3、碳酸鉀需要量------------------------------- 39
4、18-crown-6 ether需要量--------------------- 41
5、反應溫度與反應時間------------------------- 43
八、分析法之選擇性-------------------------------- 47
九、建立trans, trans-muconic acid檢量線----------- 49
十、trans, trans-muconic acid衍生物安定性之探討--- 51
十一、應用分析-------------------------------------- 52
第四章、結論------------------------------------------ 57
第五章、參考文獻-------------------------------------- 58

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