Magnetic-nanobead-based competitive enzyme-linked aptamer assay for the analysis of oxytetracycline in food
文献类型: 外文期刊
第一作者: Lu, Chunxia
作者: Lu, Chunxia;Tang, Zonggui;Kang, Lichao;Sun, Fengxia;Lu, Chunxia;Tang, Zonggui;Kang, Lichao;Sun, Fengxia;Liu, Changbin
作者机构:
关键词: Oxytetracycline;Aptamer;Nucleic acids;Enzyme-linked aptamer assay;Magnetic nanoparticles
期刊名称:ANALYTICAL AND BIOANALYTICAL CHEMISTRY ( 影响因子:4.142; 五年影响因子:3.863 )
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收录情况: SCI
摘要: This study presents a novel analytical method for the detection of oxytetracycline (OTC) in complex food matrices based on a direct competitive enzyme-linked aptamer assay and magnetic separation technology. In this protocol, free OTC competed with horseradish peroxidase labeled OTC (OTC-HRP) for binding to the OTC aptamer immobilized on magnetic beads. The parameters that can affect the response, such as avidin concentration, aptamer concentration, OTC-HRP concentration, incubation temperature, incubation time, blocking agent, and binding buffer, were optimized. Under the optimal conditions, the linear range for the OTC concentration detection is 0.5-100 ng mL(-1), with a concentration of OTC needed to obtain 50 % of the maximum signal of 14.47 ng mL(-1). The limit of detection and the limit of quantitation were 0.88 and 3.40 ng mL(-1), respectively. There was no obvious cross-reactivity with most of the tetracycline pesticides. The recovery rates ranged from 71.0 to 91.2 % for the food samples, including chicken, milk, and honey, and the relative standard deviation was less than 15.0 %. The proposed method was applied to measure OTC in real samples, and was validated using high-performance liquid chromatography. This method has the advantages of magnetic separation and the concentration effect of magnetic nanoparticles, the specificity of the aptamer, and the high-throughput of microtiter plates; it offers a promising approach for the screening of OTC because it is simple, rapid, highly sensitive, and has low cost.
分类号: O65
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