{"title":"开发和优化用于检测阿斯巴甜的安培双酶生物传感器","authors":"Tanaporn Tangtawewipat, S. Thanachasai","doi":"10.12982/cmjs.2023.064","DOIUrl":null,"url":null,"abstract":"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.","PeriodicalId":9884,"journal":{"name":"Chiang Mai Journal of Science","volume":null,"pages":null},"PeriodicalIF":0.6000,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development and Optimization of an Amperometric Bi-enzyme Biosensor for Aspartame Determination\",\"authors\":\"Tanaporn Tangtawewipat, S. Thanachasai\",\"doi\":\"10.12982/cmjs.2023.064\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"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.\",\"PeriodicalId\":9884,\"journal\":{\"name\":\"Chiang Mai Journal of Science\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.6000,\"publicationDate\":\"2023-11-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Chiang Mai Journal of Science\",\"FirstCategoryId\":\"103\",\"ListUrlMain\":\"https://doi.org/10.12982/cmjs.2023.064\",\"RegionNum\":4,\"RegionCategory\":\"综合性期刊\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MULTIDISCIPLINARY SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chiang Mai Journal of Science","FirstCategoryId":"103","ListUrlMain":"https://doi.org/10.12982/cmjs.2023.064","RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MULTIDISCIPLINARY SCIENCES","Score":null,"Total":0}
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.
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.