臺灣博碩士論文加值系統

本研究在於開發一個新的分析技術，使用微透析前處理採樣技術，對分析物進行濃縮前處理，配合逆相離子對層析法進行分離，及紫外光/可見光偵測器偵測，對水樣中的除草劑巴拉刈和二刈，做定量及定性的分析。
研究中就影響逆相離子對層析分離之因素，如離子對試劑、鹽類的種類及濃度、層析管柱、動相流速。及影響微透析前處理濃縮的因素，如透析灌注液的種類、濃度、pH值、灌注流速及樣品溶液的pH值、透析膜的長度、攪拌速率等進行詳盡探討。實驗結果顯示，在微透析的部分，將樣品溶液的pH值調整為6，使用20cm的透析管，以較快的攪拌速率(刻度4)對樣品攪拌，利用0.01M的1-己烷磺酸鈉溶液(pH = 3)做為透析灌注液，在5 μl/min的灌注流速下，巴拉刈、二刈的濃縮，可達到91及88倍。在HPLC層析的部分，利用組成為0.01 M的1-己烷磺酸鈉、添加0.01 M的三乙基胺及0.3 M的溴化鉀，以磷酸調整為pH = 3的動相溶液，在流速1.0 ml/min下，可得到相當理想的分析時間(6.4 min)及分離效果(解析度為4.3)。並且，巴拉刈、二刈的偵測極限，直接注入進行層析為1.6、1.2 ppb，經微透析濃縮後的偵測極限為0.053、0.050 ppb。

The solid phase extraction is a conventional pre-treatment method for determination of paraquat and diquat in water matrix. However, it need complicated procedure and takes much time in order to concentrate trace compounds. The conventional HPLC methods for analyze those compounds waste a lot of ion pair reagent and suffer a worse separation.
In this study, a reversed-phase ion pair liquid chromatography coupling to microdialysis method has been developed to analyze paraquat and diquat in water matrix. Conditions for obtaining optimum dialysis efficiency such as flow-rate of perfusion stream, pH in sample solution and in perfusion stream, ion-pair reagent, added-salt in the sample solution, as well as chromatographic parameters were investigated.
The experiments results indicated that the dialysis achieved at sample matrix of pH 6, and the perfusion with 0.01M sodium 1-Hexanesilfonate (pH 3, flow rate 5μl/min) offered the optimum dialysis efficiency. A reverse-phase C-18 column was used to separate the paraquat and diquat with an eluent of 0.01M sodium 1-hexanesilfonate, 0.01M triethylamine and 0.3M potassium bromide at pH 3. The measurement of analytes method was carried out with an UV detector operated at 287nm.The enrichment reached more than 100 times with RSD less than 5.68%. The paraquat and diquat compounds in waste water were well-separated with a reversed-phase ion pair liquid chromatography coupling to micro-dialysis method. It indicated the proposed method, provided a very simple, fast and good recovery method for the determination of paraquat and diquat in water matrix.

謝誌 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ⅰ
摘要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ⅱ
英文摘要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ⅲ
目錄. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ⅳ
圖次. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ⅸ
表次. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ⅩⅡ
簡語表. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ⅩⅢ
第一章 緒論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
一、前言. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
二、巴拉刈、二刈簡介. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
(一) 用途. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
(二) 毒性. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
(三) 性質. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
(四) 刈類化合物的基本資料. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
(五) 巴拉刈中毒作用機制. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7三、文獻回顧─巴拉刈、二刈的分析方法. . . . . . . . . . . . . . . . . . . . . . . 8
(一) 分光光度計法. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
(二) 固相萃取與高效液相層析/紫外光偵測器法. . . . . . . . . . . . . 8
(三) 其他分析方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
1. SPE/HPLC/LC-MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 2. MEKC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
3. SPE/LC-MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
4. SPE/CE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
5. HPLC/UV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
四、研究目的 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
五、逆相離子對層析分析法的介紹. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
六、微透析分析技術的介紹. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
(一) 透析理論的介紹. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
(二) 微透析的回收率. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
(三) 微透析技術的應用. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
第二章 研究方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
一、實驗藥品、器材、儀器設備. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
(一) 藥品. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
(二) 器材. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
(三) 儀器設備. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
二、藥品配製. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
(一) HPLC移動相(mobile phase)溶液的配製. . . . . . . . . . . . . . . . 42
(二) 儲備溶液的配製. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
(三) 標準溶液的配製. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
(四) 透析灌注液的配製. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
三、儀器操作條件. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
1. 高效能液相層析儀系統. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
2. 微透析系統. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
四、分析系統的建立. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
(一) 離子對層析分析方法的建立. . . . . . . . . . . . . . . . . . . . . . . . . . 46
(二) 透析膜的條件. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
(三) 微透析分析方法的建立. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
(四) 校正曲線的建立 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
(五) 偵測極限的測定. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
第三章 結果與討論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
一、液相層析方法的建立. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
1. UV偵測波長的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
2. 層析管柱的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
3. 離子對試劑的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
4. 動相溶液pH值的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
5. 動相中所添加之鹽類種類的選擇. . . . . . . . . . . . . . . . . . . . . . 67
6. 動相中鹽類濃度的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
7. 動相中三乙基胺濃度的影響. . . . . . . . . . . . . . . . . . . . . . . . . . 75
8. 動相中離子對試劑濃度的影響. . . . . . . . . . . . . . . . . . . . . . . . 78
9. 動相流速的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
10. 層析方法的最佳條件. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
11. 層析方法的再現性. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
二、微透析方法的建立. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
1. 透析灌注液的選擇. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
2. 透析灌注液pH值的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
3. 透析灌注液濃度的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
4. 樣品pH值的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
5. 攪拌速率的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
6. 透析管長度的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
7. 透析灌注液流速的影響. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
8. 鹽類對透析結果的影響 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
9. 微透析的最佳化條件. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
三、校正取線的建立. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
四、真實樣品的分析. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112
第四章 結論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
第五章 參考文獻. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

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