水中氨氮的測定對養殖池水質管理非常重要，因水產動物有80% ~ 90%以上含氮廢物均是以氨氮形式排泄，而氨氮是造成魚類死亡的主要原因之一。靛藍法 ( indophenol blue ) 是目前水中氨氮測定方法中最廣被接受及使用者，此法係根據Berthelot 反應( Berthelot, 1859 )，其原理為：在鹼性環境 ( pH > 9.8 ) 及在sodium nitroprusside的催化下，氨經次氯酸氧化成氯化物後再與酚反應形成靛藍色的Indophenol化合物，此外為防止因高pH而產生金屬沉澱，故於反應中亦加入檸檬酸鈉來螯合這些金屬。
然而靛藍法因反應慢，呈色不穩定，且易受溫度、pH及鹽度干擾，常導致測定結果的準確度及精確度無法掌握。過去雖有不少學者提出改善修正的方法，但均侷限於自然水域的測定，對於應用在養殖上的測定問題卻少有探討及解決。養殖上氨氮測定的特色為：試水的鹽度變化可涵蓋整個海、淡水範圍，及氨氮的濃度高且範圍大。據經驗顯示，靛藍法因在海、淡水中的反應速率及吸光係數大不相同，因而在比較不同鹽度的水中氨濃度時，其結果常因鹽度及pH干擾及因稀釋的誤差而不易判讀，此問題在快速自動分析 ( 通常為不完全反應下之測定 ) 更因加上呈色反應慢之限制而更顯嚴重。
為有系統地探討靛藍法在養殖上的測定問題及謀求改善，本研究因此以Strickland and Parsons ( 1972 ) 法 ( SP法 ) 為基礎依序分三個部份進行。
第一部份實驗：比較海水及淡水在不同最終pH條件下對靛藍法吸收光譜的影響。以其經海、淡水實驗後最佳吸收波長及吸光值的主要因素是最終pH而非鹽度，且海、淡水均適用的最佳吸收波長為630nm，最佳pH為10.5。同時，由本實驗亦可發現海水中的鹽度干擾實際上是因反應的最終pH不同所造成的，且經由最佳模式化結果，顯示當pH在9.8~10.5時吸光係數與最終pH有良好的二次模式關係(R2>98%)，似可用來校正SP法測定結果因最終pH不同而造成的差異 ( 數學模式修正法 )。
第二部分：比較靛藍法在不同海水比例下的NaOH滴定曲線及緩衝容量，以探討海水鹽度對最終pH的影響，並求出不同海水比例在最佳pH下所需的NaOH最佳添加量。由結果顯示，NaOH滴定曲線隨海水比例不同而異，經比對結果與理論滴定曲線並無顯著差異。因此顯然可知，海水比例不同會影響靛藍法的最終pH，更確定鹽度干擾效應來自於最終pH不同。且由本實驗滴定曲線所求得之NaOH最佳添加量，似可用於試劑的調整以抵銷所謂的〝鹽度干擾〞的問題( 試劑修正法 )。
第三部份：以標準曲線及實際樣品比較SP法、數學模式修正法及試劑修正法對6種不同比例海水樣品的測定效果。由標準曲線結果顯示：海水比例會影響SP法的標準曲線斜率，對試劑修正法及數學模式修正法的標準曲線則無影響。經由養殖池實際樣品測定的驗證結果顯示：SP法的最終pH值隨鹽度增加而減少，試劑修正法的最終pH則不受鹽度影響而定數在10.5左右，數學模式修正法的最終pH值則固定在10.5。另經卡方試驗結果顯示數學模式修正法確實與試劑修正法的符合性較高。
由本研究結果可確實證明靛藍法測氨的鹽度干擾問題事實上是因反應最終pH不同所造成的。同時，亦可得知靛藍法最佳吸收波長為630nm，最佳pH為10.5。因此，若能如本研究的試劑修正法依試水滴定曲線調整試劑NaOH用量，將最終pH定數在10.5左右，即可去除鹽度干擾，進而得到不受海水或淡水影響的測定結果，且對高濃度試水的測定而言，亦不會因稀釋造成鹽度改變而帶來誤差。

第一章 前言 1
第二章 比較海水及淡水在不同最終pH條件下對靛藍法吸收光譜的影響 3
摘要 3
1.前言 4
2.材料與方法 6
3.結果 8
4.討論 10
第三章 比較靛藍法在不同海水比例下的NaOH滴定曲線及緩衝容量 12
摘要 12
1.前言 13
2.材料與方法 15
2.1實驗上的滴定曲線及緩衝容量 15
2.1.1試劑材料準備 15
2.2理論上的滴定曲線及緩衝容量 17
2.2.1理論上滴定弱酸模擬流程 17
2.2.2理論上的滴定曲線公式 18
2.3更改試劑於不同比例海水之配方及測氨步驟 23
2.3.1 NaOH的添加量 22
2.4卡方測驗之符合性測驗 23
3.結果 24
4.討論 26
第四章 以SP法、數學模式修正法、以及試劑修正法比較不同
鹽度之不同水源之氨濃度 31
摘要 31
1.前言 32
2.材料與方法 34
3.結果 39
4.討論 43
第五章 總結 46
參考文獻 47
表 53
圖 62
個人簡歷 77

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