葡萄糖/尿素碳六十固定化酵素選擇性電極之研製與應用

Preparation and Application of Fullerene C60 Immobilized Enzyme Electrodes for Glucose and Urea

張祐穎(Yu-Ying Chang)

指導教授 ： 施正雄

國立臺灣師範大學/理學院/化學系/碩士(2002年)

摘要

中文摘要
本研究合成固定化碳六十/葡萄糖氧化酵素(C60 －Glucose Oxidase)和固定化碳六十/尿素水解酵素(C60-Urease)並分別應用在葡萄糖及尿素選擇性C60-cryptand[2,2] / PVC電極感測系統中，用以偵測待測溶液中的葡萄糖或尿素分子。在C60-cryptand[2,2]葡萄糖薄膜電極系統中，偵測葡萄糖分子被葡萄糖氧化酵素催化為葡萄糖酸在經解離得到的氫離子，而C60-cryptand[2,2]尿素選擇性薄膜電極中，偵測水溶液中尿素分子經尿素水解酵素催化所形成的銨根離子。實驗中將分別探討碳六十固定化酵素的含量、溫度、pH值的改變及干擾因子對電極電位變化的影響。在葡萄糖選擇性薄膜電極偵測系統中，固定化C60 －Glucose Oxidase在30~35oC的溫度下，有很好的催化效果，而最適pH值則在6~7之間，且發現當膜上所含C60 －Glucose Oxidase的量增加時，催化的效果越明顯。而pH值的效應上發現，當待測溶液的pH值在6~7之間時，有最好的催化效果。此選擇性電極在葡萄糖分子濃度為10-4~10-1 M之間，有52.5mV/decade的線性電極電位反應，其穩定性及再現性都不錯。在干擾係數的測定方面，整體的選擇性係數在10-2左右，如鹼金族、鹼土族及過渡金屬。在尿素選擇性薄膜電極偵測系統中，最適pH值也在6~7之間而最佳溫度為室溫。此電極在尿素分子濃度為10-4~10-1 M之間，有51.7mV/decade的線性電極電位反應。在干擾離子方面，大部分干擾五種的選擇性係數也在10-2左右，如鹼金族、鹼土族及過渡金屬離子。

關鍵字

葡萄糖 ； 尿素 ； 葡萄糖氧化酵素 ； 尿素水解酵素 ； 固定化酵素 ； 選擇性電極

並列摘要

Abstract
Immobilized C60 －Glucose Oxidase and C60-Urease were synthesized and applied in glucose and urea C60-cryptand[2,2] / PVC electrodes to detect glucose and urea in aqueous solutions. The C60-cryptand[2,2] glucose selective membrane electrode detection system to detect the H+ ion which is the product of the gluconic acid dissociation catalyzed by C60-glucose oxidase in another PVC membrane and C60-cryptand[2,2] urea selective membrane electrode detection system to detect the NH4+ which is the production of urea hydrolysis catalyzed by C60-urease in another PVC membrane. The effects of mass of immobilized enzymes, temperature, pH value and interference ions on the potential response were investigated and discussed. For glucose selective membrane electrode detection system, it was found that the catalytic ability of immobilized C60-glucose oxidase showed an optimum temperature at 30℃~35℃ and optimum pH between 6~7, the potential response showed increase with the increased mass of immobilized C60-glucose oxidase in PVC membrane. This electrode gave a linear response 52.3mV/decade within the concentration range in 10-4~10-1 M of glucose and showed selectivity coefficients around 10-2 for most interfering ions, e.g., alkali, alkaline earth and transition metal ions. For urea selective membrane electrode detection system, the optimum is at testing solution pH value 6~7, and optimum temperature at room temperature. This urea electrode also exhibited a linear response 51.7 mV/decade within the concentration range in 10-4~10-1 M of urea and also showed selectivity coefficients around 10-2 for most interfering ions, e.g., alkali, alkaline earth and transition metal ions.

並列關鍵字

glucose ； glucose oxidase ； urea ； urease ； immobilized enzyme ； selective electrode