二價銅對高效液相層析搭配2,4-二硝基苯肼管柱前衍生系統分析脂肪醛的影響與探討

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研究生：	葉雲臺
論文名稱：	二價銅對高效液相層析搭配2,4-二硝基苯肼管柱前衍生系統分析脂肪醛的影響與探討 Effects of Copper(II) on the Determination of Aliphatic Aldehydes by HPLC Using DNPH Precolunm Derivatization
指導教授：	吳劍侯
口試委員:
學位類別：	碩士 Master
系所名稱：	原子科學院 - 生醫工程與環境科學系 Department of Biomedical Engineering and Environmental Sciences
論文出版年：	2009
畢業學年度：	97
語文別：	中文
論文頁數：	43
中文關鍵詞：	二價銅、2,4-二硝基苯肼、斐林反應、醛
外文關鍵詞：	Cu(II), 2,4-dinitrophenylhydrazine(DNPH), Fehling reaction, aldehyde
相關次數：	點閱：89 下載：0
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利用2,4-二硝基苯肼(2,4-dinitrophenylhydrazine, DNPH)做為量測醛類的衍生試劑為最常見及廣泛應用的分析方法，通常在酸性催化衍生下以高效率液相層析儀 (high performance liquid chromatography, HPLC)搭配紫外光可見光偵測器(UV/Vis detector)量測在360 nm處DNPH衍生物(hydrazone)的光譜吸收；我們藉由探討不同溫度下衍生反應的平衡常數，及衍生最佳pH值與DNPH質子化的關係對此衍生反應的機制有更進一步的了解，發現此衍生反應在室溫下有最高的反應平衡常數，DNPH質子化的平衡常數K1= 46.8 M-1。過去的研究發現二價銅（Cu(II)）易與DNPH上的聯氨(Hydrazine)反應，進而造成分析方法的干擾，然而Cu(II)對此衍生反應的影響除了對DNPH造成分解外，Cu(II)與醛類的斐林反應(Fehling reaction)及Cu(II)對衍生物穩定性干擾，也都為影響此反應的因素，本篇研究目的為釐清Cu(II)對衍生反應造成的影響由何者所貢獻，並藉由加入EDTA(Ethylene dinitrilotrteaacetic acid) 做為Cu(II)的抑制劑，發現在加入適量的EDTA情況下可以有效降低Cu(II)對此反應的干擾。

目錄
中文摘要.........................................................................................................................I
英文摘要.......................................................................................................................II
謝誌..............................................................................................................................III
目錄..............................................................................................................................IV
圖目錄..........................................................................................................................VI
表目錄........................................................................................................................VII

第一章    前言................................................................................................................1
1.1    簡介..................................................................................................................1
1.2    研究目的及動機..............................................................................................2
第二章    文獻回顧......................................................................................................3
2.1    醛的測量方法….................................................................................………3
2.1.1.    直接進樣法....................................................………............................3
2.1.2.    衍生法........................ ...............................................................………3
2.2    銅的介紹............................ ....................................................…........………6
2.3    醛與DNPH........ ....................................................……....................………7
2.4    DNPH的反應............................ ....................................................…………9
2.5    DNPH的應用............................... ....................................................………11
2.6    斐林反應. ....................................................................................………12
第三章    實驗方法......................................................................................................13
3.1    實驗裝置.........................................................................................................13
3.1.1.    紫外光可見光譜儀（UV/Vis spectrometer）.....................................13
3.1.2.    高效率液相層析儀（HPLC）..................................... ..........................13
3.1.3.    其他儀器..............................................................................................14
3.2    實驗計算軟體（MINTEQ）............................................................................14
3.3    實驗藥品..................................... ...................................................................15
3.2.1.    藥品..................................... ... ............................................................15
3.2.2.    母液配置..................................... ............................ ...........................16
3.4    分析流程........................................ .......................... .....................................18
3.4.1.    以UV/Vis觀察DNPH及DNPH衍生物之光譜特性......................18
3.4.2.    醛的分析..................................... ........................................................18
3.4.3.    酸的分析................................. ...................... .....................................18
3.4.4.    斐林反應的測試..................................... .........................................19
3.4.5.    實驗流程圖..................................... ..................................... ..............19
第四章    結果與討論…..............................................................................................20
4.1    衍生反應的最佳化..................................... ........................ ..........................20
4.1.1.    HPLC分析條件..................................... .............................................20
4.1.2.    溫度對衍生反應的影響化................................... ..............................21
4.1.3.    反應平衡常數的探討..........................................................................23
4.1.4.    DNPH質子化與衍生pH值................................................................26
4.1.5.    Cu(II)及EDTA的影響.....................................................................29
4.1.6.    衍生條件最佳化..................................... .......................... .................32
4.1.7.    反應機制的探討...............................................................................33
4.2    斐林反應的測試..................................... ........... ......................................34
4.3    衍生物的穩定性測試..................................... ....... .......................................35
第五章    結論與展望..………………………………………..……………………..38
參考文獻......................................................................................................................39
附錄I    醛之檢量線............................................................ ........................................i
附錄II    各個醛在不同溫度下反應常數....................................................................ii
附錄III    各個醛在不同pH值下改變Cu(II)、EDTA濃度及衍生時間……….........iii

                                

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