Development of Cu(II)/Cu(I)-induced quantum dot-mediated fluorescence immunoassay for the sensitive determination of eth
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ORIGINAL PAPER
Development of Cu(II)/Cu(I)-induced quantum dot-mediated fluorescence immunoassay for the sensitive determination of ethyl carbamate Kai Zhou 1,2 & Zhi-Long Wang 3 & Lin Luo 1 & Yong-Zhen Dong 3 & Jin-Yi Yang 1 & Hong-Tao Lei 1 & Hong Wang 1 & Yu-Dong Shen 1 & Zhen-Lin Xu 1 Received: 24 March 2020 / Accepted: 18 August 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A series of haptens were rationally designed for producing monoclonal antibodies specific for EC and a simple fluorescence immunoassay platform was developed for the sensitive determination of EC based on alkaline phosphatase (ALP)triggered Cu+ quenching of CdSe quantum dots (QDs). It was noted that Cd as a fluorescence substrate in CdSe QDs can be selectively substituted by Cu+ that resulted in a more significant fluorescence quenching in comparison with Cu2+. Meanwhile, because ALP catalyzed ascorbic acid phosphate and then assisted the transformation of Cu2+ to Cu+, the change in fluorescence intensity was found to be proportional to ALP concentration. After simple magnetic separation, the sensitivity and linear range of the established assay were improved approximately 53-fold and an order of magnitude, respectively, when compared with the conventional ELISA. The proposed platform was able to both amplify the signal and eliminate matrix interferences, making it a promising to determine EC as well as other contaminants in complex food matrix in a highly sensitive and simple manner. Keywords Ethyl carbamate . Monoclonal antibody . Fluorescence immunoassay . Quantum dots
Introduction Ethyl carbamate (EC) is a group 2A carcinogen extensively occurring in alcoholic beverages and is normally formed as a by-product during fermentation and sterilization process [1, 2]. Concerning the wide occurrence and potential toxicity to humans, many countries have established criteria for EC in Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04502-5) contains supplementary material, which is available to authorized users. * Zhen-Lin Xu [email protected] 1
Guangdong Provincial Key Laboratory of Food Quality and Safety, Guangdong Laboratory of Lingnan Modern Agriculture, College of Food Science, South China Agricultural University, Wushan Street No. 483, Guangzhou 510642, People’s Republic of China
2
College of Pharmacy and Life Science, Jiujiang University, Jiujiang 332000, People’s Republic of China
3
College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China
different beverages varying from 15 to 1000 μg/L. For example, the Canada government set the MRL for EC in wine, brand, distilled spirits, fortified spirits, and sake as 30, 400, 150, 100, and 200 μg/L [3]. Presently, the monitoring of EC and the prevention of its accumulation have caused widespread concern [4]. Therefore, it is of great value to develop a high-precision, low analytic limitation, easy-to-use detection method for EC supervision. Compared w
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