呋喃它酮代谢物可视化凝胶柱快速免疫检测方法研究

Study of a Gel-based Immunoassay Method for the Rapid Visual Detection of Furaltadone Metabolites

摘要:

本研究建立了兽药呋喃它酮代谢物（Furaltadone metabolites，AMOZ）的免疫亲和凝胶柱检测新方法，并用于动物源性食品中呋喃它酮代谢物残留的可视化快速检测。整个检测在一个标准的1 mL固相萃取（SPE）柱中进行，柱中包含两层：一个结合了呋喃它酮代谢物特异性抗体的凝胶检测层和一个结合了辣根过氧化物酶（HRP）抗体的凝胶质控层。将样品提取液与酶标抗原预混合后加入到检测柱中，基于抗原抗体的竞争结合反应和辣根过氧化物酶的酶促反应，根据检测柱的检测层颜色的深浅和有无来定性，半定量检测呋喃它酮代谢物的含量。整个检测过程可以在10 min内完成，该凝胶检测柱的检测限为20 μg/L；牛肉、鲷鱼、鸡肉、虾肉、猪肉、鱿鱼、鸡肝和黄花鱼等动物源性食品中呋喃它酮代谢物的检测限为3 μg/kg。该方法准确度高、特异性好、检测步骤简单，适用于大量样品中呋喃它酮代谢物残留的快速检测。

Abstract:

A new method using an immuno-affinity column for the detection of metabolites of the veterinary drug furaltadone (AMOZ) was established in this study, and was applied for the rapid visual detection of AMOZ residues in animal-derived foods. Measurement was conducted in a standard solid phase extraction cartridge (1 mL) that consisted of a detection layer, containing anti-furaltadone metabolite antibody-coupled gel, and a quality control layer with anti-HRP antibody-coupled gel. The sample extract was pre-mixed with the enzyme-labeled antigen and the mixture was added to the detection column. Based on the direct competitive immuno-reaction and the HRP enzymatic reaction, qualitative analysis was performed according to the presence and strength of color, and the content of AMOZ was determined semi-quantitatively. The whole procedure was completed within 10 min, and the limit of detection of the gel-based detection column for the detection of furaltadone metabolite derivatives (NPAMOZ) was 20 μg/L. The visual detection limits of this method for the detection of AMOZ in animal-derived foods (beef, snapper, chicken, shrimp, pork, squid, chicken liver, and yellow croaker) were 3 μg/kg. This gel-based visual immunoassay method has high accuracy and good specificity and is easy to use; it is suitable for the rapid detection of AMOZ residues in a large number of animal-derived food samples.