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研究生:張春單
研究生(外文):CHUN-TAN CHANG
論文名稱:以微透析/HPLC-UV測定廢水中Aniline和2-Chloroaniline及乳品中有機酸之研究
指導教授:鄭政峰鄭政峰引用關係
指導教授(外文):JEN-FON JEN
學位類別:碩士
校院名稱:國立中興大學
系所名稱:化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:97
中文關鍵詞:微透析技術高效能液相層析有機酸
外文關鍵詞:micro-dialysis2-chloroanilineanilineorganic acidshigh performance liquid chromatography
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本研究將微透析技術應用於環境分析及乳品中成分分析,以微透析作為樣品前處理步驟,配合高效能液相層析分離及紫外光偵測系統,同步分離與測定高分子工業廢水中aniline和2-chloroaniline及發酵乳品中有機酸。研究中依分析物之特性,探討樣品溶液中的pH值和鹽類濃度、透析灌注液中極性修飾劑濃度及流速等影響透析之因素,尋找最佳條件,配合HPLC/UV系統之分析測定。
研究結果顯示,對於高分子工業廢水中苯胺之測定以逆相 C-18為層析管柱,40 %甲醇水溶液(pH 7.0),流速為1ml / min之沖提液,可得最佳的分析結果。對於發酵乳品中有機酸之分析,則以同樣分離管,pH值2.9 的水為沖提液,在流速為0.6 ml / min沖提下,可得最佳的分析結果。由實際樣品分析結果顯示,以微透析連接高效能液相層析之技術來分析高分子工業廢水中aniline和2-chloroaniline及發酵乳品中有機酸,是個快速、簡單的方法。
Determination of aniline and 2-chloroaniline in polymer industrial wastewater and organic acids in fermentation products of milk are examined with high performance liquid chromatography on-lined micro-dialysis. The optimum conditions for obtaining dialysis efficiency such as flow-rate and polarity modifier in perfusion stream, pH and added-salt in sample solution, as well as chromatographic conditions were investigated.
Experimental results indicated that the aniline and 2-chloroaniline are well separated in an acceptable time on a reversed-phase C-18 column eluted with 40% aqueous methanol solution at pH 7.0 and 1.0 ml/min flow-rate. The organic acids (citric acid, lactic acid, acetic acid, and formic acid) can be well separated with water at pH 2.9 and 0.6 ml/min flow-rate. The proposed method provided a very simple procedure to determine aniline and 2-chloroaniline in wastewater and organic acids in fermentation products of milk. Application was illustrated by the analysis of aniline and 2-chloroaninine in wastewater released from a polymer factory and organic acids in two fermentation products.
目 錄
摘要‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧Ⅰ
英文摘要(Abstract)‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧Ⅱ
謝誌‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧Ⅲ
目錄‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧Ⅳ
圖次‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧Ⅷ
表次‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧IV
簡語表‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧IV
第一章 緒論‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧01
一、研究動機‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧01
二、Aniline和2-Chloroaniline之概述‧‧‧‧‧‧‧‧‧ ‧02
三、乳品中常見有機酸之概述‧‧‧‧‧‧‧‧‧‧‧‧‧‧05
四、微透析分析方法概述‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧07
A、微透析之理論介紹‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧08
B、微透析技術之應用‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧16
第二章 研究方法‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧17
一、實驗藥品、器材、儀器設備‧‧‧‧‧‧‧‧‧‧‧‧‧17
A.藥品‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧17
B.器材‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧18
C.儀器設備‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧19
二、藥品配製‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧20
三、儀器操作條件‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧30
A. 高效能液相層析儀 (HPLC) 系統‧‧‧‧‧‧‧‧‧‧‧30
B. 微透析 (microdialysis) 系統‧‧‧‧‧‧‧‧‧‧‧‧31
四、實驗過程‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧33
A. 透析薄膜條件的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧33
B. 液相層析方法的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧35
C. 微透析分析方法的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧36
五、校正曲線的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧38
六、偵測極限的測定‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧39
第三章 結果與討論‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧40
一、Aniline和2-Chloroaniline分析方法的建立‧‧‧‧‧‧40
A、液相層析方法的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧40
1.UV偵測波長的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧40
2.層析管柱的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧40
3.動相的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧42
B、微透析系統方法之建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧46
1.樣品溶液pH值的影響‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧46
2. 基質效應的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧48
3.透析灌柱液中Methanol濃度的影響‧‧‧‧‧‧‧‧‧‧‧50
4. 透析流速的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧50
5. 透析暨層析方法再現性的測試‧‧‧‧‧‧‧‧‧‧‧‧‧54
C、校正曲線的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧54
D、透析回收率的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧59
E、真實樣品的分析‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧59
三、四種有機酸分析方法的建立‧‧‧‧‧‧‧‧‧‧‧‧‧62
A. 液相層析分析方法的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧62
1.UV偵測波長的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧62
2.層析管柱的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧62
3.動相的選擇‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧64
B、微透析系統方法之建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧66
1. 樣品溶液pH值的影響‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧66
2. 基質效應的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧68
3. 透析流速的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧68
4. 透析暨層析方法再現性的測試‧‧‧‧‧‧‧‧‧‧‧‧72
C、校正曲線的建立‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧72
D、透析回收率的探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧79
E、真實樣品的分析‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧79
第四章 結論‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧86
第五章 參考文獻‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧87
圖 次
圖(一)各種透析薄膜式樣圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧09
圖(二)醋酸纖維之化學結構式‧‧‧‧‧‧‧‧‧‧‧‧‧12
圖(三)儀器裝置圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧21
圖(四)中空透析薄膜製作完成圖‧‧‧‧‧‧‧‧‧‧‧34
圖 (五) Aniline、2-chloroaniline之UV吸收光譜‧‧‧‧41
圖(六)沖提液中不同pH值對aniline、2-chloroaniline吸收面積之關係圖‧‧‧‧‧‧‧‧‧‧‧‧‧ ‧‧‧‧‧‧‧‧‧‧‧‧43
圖(七)沖提液中CH3OH濃度對滯留時間之關係‧‧‧‧‧‧44
圖(八)各2 ppm Aniline和2-Chloroaniline標準溶液之層析圖譜‧45
圖(九)樣品溶液中不同pH值對透析回收率之探討‧‧‧‧‧47
圖(十)樣品溶液中添加不同濃度的NaCl對透析回收率之探討‧49
圖(十一)灌注液中CH3OH濃度對回收率之探討‧‧‧‧‧‧51
圖(十二)灌注液中不同流速對透析回收率之探討‧‧‧‧‧52
圖(十三)Aniline直接注入HPLC系統之校正曲線圖‧‧‧‧56
圖(十四)2-Chloroaniline直接注入HPLC系統之校正曲線圖‧56
圖(十五)Aniline以10ml/min透析流速連線方式進入HPLC系統之校正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧57
圖(十六)2-Chloroaniline以10ml/min透析流速連線方式進入HPLC系統之校正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧57
圖(十七)Aniline和2-chloroaniline各2ppm之真實樣品層析圖‧60
圖(十八)有機酸之UV吸收光譜圖‧‧‧‧‧‧‧‧‧‧‧‧63
圖(十九)各100 ppm的有機酸標準溶液之層析圖譜‧‧‧‧65
圖(二十)樣品溶液中不同pH值對透析面積之探討‧‧‧‧‧67
圖(二十一)樣品溶液中添加不同濃度的NaCl對透析面積之探討‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧69
圖(二十二)灌注液中不同流速對透析面積之探討‧‧‧‧‧70
圖(二十三)Acetic acid直接注入HPLC系統之校正曲線圖‧‧‧75
圖(二十四)Acetic acid以10ml/min透析流速連線方式進入HPLC系統之校正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧75
圖(二十五)Lactic acid直接注入HPLC系統之校正曲線圖‧‧76
圖(二十六)Lactic acid以10ml/min透析流速連線方式進入HPLC系統之校正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧76
圖(二十七)Formic acid直接注入HPLC系統之校正曲線圖‧77
圖(二十八)Formic acid以10ml/min透析流速連線方式進入HPLC系統之校正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧77
圖(二十九)Citric acid直接注入HPLC系統之校正曲線圖‧‧78
圖(三十)Citric acid以10ml/min透析流速連線方式進入HPLC系統之校
正曲線圖‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧78
圖(三十一)真實樣品之HPLC層析圖(a)優酪乳(b)養樂多‧82
圖(三十二)養樂多在不同時間的微透析系統下之濃度變化關係圖‧84
圖(三十三)優酪乳在不同時間的微透析系統下之濃度變化關係圖‧85
表 次
表(一)Aniline的基本基料‧‧‧‧‧‧‧‧‧‧‧‧‧‧03
表(二)2-Chloroaniline的基本基料‧‧‧‧‧‧‧‧‧‧‧04
表(三)四種有機酸的基本基料‧‧‧‧‧‧‧‧‧‧‧‧‧06
表(四)各種透析薄膜材質的種類‧‧‧‧‧‧‧‧‧‧‧‧13
表(五)苯胺HPLC系統之最佳層析操作條件‧‧‧‧‧‧‧‧46
表(六)苯胺透析系統之最佳操作條件‧‧‧‧‧‧‧‧‧‧53
表(七)Aniline透析暨層析方法再現性測試‧‧‧‧‧‧‧55
表(八)2-Chloroaniline透析暨層析方法再現性測試‧‧‧55
表(九)Aniline和2-chloroaniline之檢量線參數‧‧‧‧‧ 58
表(十)真實樣品之分析‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧61
表(十一)有機酸HPLC系統之最佳層析操作條件‧‧‧‧‧‧64
表(十二)有機酸透析系統之最佳操作條件‧‧‧‧‧‧‧‧71
表(十三)Formic acid透析暨層析方法再現性測試‧‧‧‧73
表(十四)Lactic acid透析暨層析方法再現性測試‧‧‧‧73
表(十五)Acetic acid透析暨層析方法再現性測試‧‧‧‧74
表(十六)Citric acid透析暨層析方法再現性測試‧‧‧‧74
表(十七)四種有機酸之檢量線參數‧‧‧‧‧‧‧‧‧‧‧ 80
表(十八)真實樣品之分析結果‧‧‧‧‧‧‧‧‧‧‧‧‧81
表(十九)養樂多之分析‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧83
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