开发和优化用于检测阿斯巴甜的安培双酶生物传感器

IF 0.6 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES Chiang Mai Journal of Science Pub Date : 2023-11-30 DOI:10.12982/cmjs.2023.064
Tanaporn Tangtawewipat, S. Thanachasai
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引用次数: 0

摘要

开发了一种基于α-糜蛋白酶(CHY)和酒精氧化酶(AOX)的安培双酶生物传感器,用于在流动注射分析(FIA)系统中检测阿斯巴甜。利用吸附技术将 CHY 和 AOX 分别固定在微珠载体上。阿斯巴甜在 CHY 催化下裂解产生甲醇,甲醇在 AOX 催化下转化为甲醛和过氧化氢。形成的过氧化氢通过铂电极进行安培检测。在酶固定和操作条件方面对生物传感器的性能进行了优化。最合适的酶固定条件为 250 U/mL CHY 和 100 U/mL AOX,固定时间为 60 分钟。所开发的天冬酰胺生物传感器的最佳操作条件是:流速为 0.5 mL/min,pH 值为 8.0,对 Ag/AgCl 的施加电位为 +0.70 V。在最佳条件下,所开发的生物传感器在阿斯巴甜浓度为 0.15-1.0 mM 的范围内呈线性响应(测定系数 (R2) = 0.9941),灵敏度为 5.52 μA/mM-cm2,检测限为 0.10 mM。所开发的生物传感器成功地应用于商业样品中阿斯巴甜的测定。
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Development and Optimization of an Amperometric Bi-enzyme Biosensor for Aspartame Determination
An amperometric bi-enzyme biosensor based on α-chymotrypsin (CHY) and alcohol oxidase (AOX) was developed for detection of aspartame in a flow injection analysis (FIA) system. The CHY and AOX were separately immobilized on bead supports using an adsorption technique. The cleavage of aspartame was catalyzed by CHY to produce methanol that was converted by AOX to formaldehyde and hydrogen peroxide. The formed hydrogen peroxide was detected amperometrically at a platinum electrode. The biosensor performance was optimized with respect to enzyme immobilization and operating conditions. The most suitable conditions for enzyme immobilization were 250 U/mL CHY and 100 U/mL AOX with 60 min immobilization time. The optimal conditions for operating the developed aspartame biosensor were a flow rate of 0.5 mL/min and pH 8.0 at an applied potential of +0.70 V versus Ag/AgCl. Under the optimal conditions, the developed biosensor showed a linear response over the aspartame concentration range 0.15–1.0 mM (coefficient of determination (R2) = 0.9941) with a sensitivity of 5.52 μA/mM·cm2 and a detection limit of 0.10 mM. The developed biosensor was successfully applied in the determination of aspartame in commercial samples.
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来源期刊
Chiang Mai Journal of Science
Chiang Mai Journal of Science MULTIDISCIPLINARY SCIENCES-
CiteScore
1.00
自引率
25.00%
发文量
103
审稿时长
3 months
期刊介绍: The Chiang Mai Journal of Science is an international English language peer-reviewed journal which is published in open access electronic format 6 times a year in January, March, May, July, September and November by the Faculty of Science, Chiang Mai University. Manuscripts in most areas of science are welcomed except in areas such as agriculture, engineering and medical science which are outside the scope of the Journal. Currently, we focus on manuscripts in biology, chemistry, physics, materials science and environmental science. Papers in mathematics statistics and computer science are also included but should be of an applied nature rather than purely theoretical. Manuscripts describing experiments on humans or animals are required to provide proof that all experiments have been carried out according to the ethical regulations of the respective institutional and/or governmental authorities and this should be clearly stated in the manuscript itself. The Editor reserves the right to reject manuscripts that fail to do so.
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