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β-激动剂快速检测技术研究:基于二聚体人工抗原的多残留荧光免疫层析法

Research on Rapid Detection Technology for -Agonists: Multi-Residue Fluorescence Immunochromatography Based on Dimeric Artificial Antigen.

作者信息

Liu Miaomiao, Ma Biao, Wang Yaping, Chen Erjing, Li Jiali, Zhang Mingzhou

机构信息

Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, China.

出版信息

Foods. 2022 Mar 18;11(6):863. doi: 10.3390/foods11060863.

Abstract

To detect two types of β2-agonist residues at the same time, we coupled two haptens of clenbuterol (CLE) and ractopamine (RAC) to the same carrier protein through diazotization to prepare dimeric artificial antigen, and a fluorescent lateral flow immunoassay method based on europium nanoparticles (EuNP-FLFIA) was established by combining polyclonal antibodies with europium nanoparticles to form probes. Under optimized conditions, the EuNP-FLFIA could simultaneously detect eight aniline-type and one phenol-type β2-agonists, and the limits of detection (LOD) were 0.11−0.19 ng/mL and 0.12 ng/mL, respectively. The recovery rate of this method was 84.00−114.00%. This method was verified by liquid chromatography−tandem mass spectrometry (LC-MS/MS), and the test results were consistent (R2 > 0.98). Therefore, the method established in this study could be used as a high-throughput screening for the efficient and sensitive detection of β2-agonists in food.

摘要

为了同时检测两种β2-激动剂残留,我们通过重氮化反应将两种克伦特罗(CLE)和莱克多巴胺(RAC)半抗原偶联到同一载体蛋白上,制备了二聚体人工抗原,并通过将多克隆抗体与铕纳米颗粒结合形成探针,建立了基于铕纳米颗粒的荧光侧向流动免疫分析方法(EuNP-FLFIA)。在优化条件下,EuNP-FLFIA能够同时检测8种苯胺型和1种酚型β2-激动剂,检测限(LOD)分别为0.11−0.19 ng/mL和0.12 ng/mL。该方法的回收率为84.00−114.00%。该方法通过液相色谱-串联质谱(LC-MS/MS)进行验证,测试结果一致(R2>0.98)。因此,本研究建立的方法可作为一种高通量筛选方法,用于高效、灵敏地检测食品中的β2-激动剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb47/8949518/4a592b7ebff4/foods-11-00863-g001.jpg

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