Copper Oxide Nanoparticle-Based Immunosensor for Zearalenone Analysis by Combining Automated Sample Pre-Processing and High-Throughput Terminal Detection.

Zhihong Xuan,Jin Ye,Li Li,Hongmei Liu,Songxue Wang,Yanxiang Wu

doi:10.3390/s21196538

Zhihong Xuan, Jin Ye + Show 4 more

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https://doi.org/10.3390/s21196538

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Abstract

A rapid and high-throughput fluorescence detection method for zearalenone (ZEN) based on a CuO nanoparticle (NP)-assisted signal amplification immunosensor was developed using an automated sample pretreatment and signal conversion system. CuO NPs with high stability and biocompatibility were used as carriers to immobilize anti-ZEN antibodies. The obtained CuO NP-anti-ZEN can maintain the ability to recognize target toxins and act as both a signal source and carrier to achieve signal conversion using automated equipment. In this process, target toxin detection is indirectly transformed to Cu2+ detection because of the large number of Cu2+ ions released from CuO NPs under acidic conditions. Finally, a simple and high-throughput fluorescence assay based on a fluorescent tripeptide molecule was employed to detect Cu2+, using a multifunctional microporous plate detector. A good linear relationship was observed between the fluorescence signal and the logarithm of ZEN concentration in the range of 16.0–1600.0 μg/kg. Additionally, excellent accuracy with a high recovery yield of 99.2–104.9% was obtained, which was concordant with the results obtained from LC-MS/MS of naturally contaminated samples. The CuO NP-based assay is a powerful and efficient screening tool for ZEN detection and can easily be modified to detect other mycotoxins.

Highlights

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by several species of Fusarium, F. graminearum, F. culmorum, F. equiseti, and F. verticillioides [1]
We developed a CuO NP-based immunosensor for ZEN detection by combining automated sample preprocessing with high-throughput terminal detection
The principle of the CuO NP-based immunosensor for ZEN detection is shown in Scheme 1

Summary

Introduction

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by several species of Fusarium, F. graminearum, F. culmorum, F. equiseti, and F. verticillioides [1]. ZEN and its derivatives have been widely detected in agricultural products, wheat, corn, barley, soybeans, and oats [2]. To protect humans from ZEN exposure and reduce economic losses, national and international agencies adopted regulations for the control of ZEN contamination. The maximum residue level of 350 μg/kg in unprocessed corn was set by the European Union (Commission Regulation, EC No 1126/2007). China has set a maximum level of 60 μg/kg for ZEN in cereal and cereal-based products (National Food Safety Standard GB2761-2017). Based on the important effects of ZEN on human and animal health, monitoring of ZEN in food and feed is a global priority for protecting customers’ health and facilitating international food and feed trade

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