Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics

文献类型： 外文期刊

第一作者： Du Xin-wei

作者： Du Xin-wei;She Yong-xin;Zhao Feng-nian;Wang Jing;Wang Shan-shan;Jin Fen;Shao Hua;Jin Mao-jun;Zheng Lu-fei;Du Xin-wei;She Yong-xin;Zhao Feng-nian;Wang Jing;Wang Shan-shan;Jin Fen;Shao Hua;Jin Mao-jun;Zheng Lu-fei;Zhang Yan-xin;Liu Guang-yang

作者机构：

关键词： dummy molecularly imprinted polymers;melamine;fluorescent competitive assay;artificial antibody

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：2.848； 五年影响因子：2.979 ）

ISSN： 2095-3119

年卷期： 2016 年 15 卷 5 期

页码：

收录情况： SCI

摘要： A fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers (DMIPs) as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased binding of fluorescent substances to DMIPs due to increased concentrations of melamine in the solutions. DMIPs for melamine were synthesized under a hot water bath in the presence of the initiator azobisisobutyronitrile (AIBN) using 2,4-diamino-6-methyl-1,3,5-triazine (DAMT) as a dummy template, methacrylic acid (MAA) as a functional monomer, and ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent. The adsorption capacity and selectivity of DMIPs for melamine were evaluated by the isothermal adsorption curve and Scatchard analysis. The evaluation results showed that the synthesized DMIPs had specific recognition sites for melamine and the maximum adsorption amount was 1066.33 mu g g(-1). Later, 5-(4,6-dichlorotriazinyl) amino fluorescein (DTAF) with a triazine ring, which slightly resembles melamine, was selected as the fluorescent substance. The fluorescent competitive assay using DMIPs as the antibody mimics was finally established by selecting and optimizing the reaction solvents, DMIPs amount, DTAF concentration, and incubation time. The optimal detection system showed a linear response within range of 0.05-40 mg L-1 and the limit of detection (LOD) was 1.23 mu g L-1. It was successfully applied to the detection of melamine in spiked milk samples with satisfactory recoveries (71.9 to 86.3%). According to the comparative analysis, the result of optimized fluorescent competitive assay revealed excellent agreement with the HPLC-MS/MS result for melamine.

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