March C, Manclús J J, Jiménez Y, Arnau A, Montoya A
Instituto de Investigación e Innovación en Bioingeniería, Universidad Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain.
Talanta. 2009 May 15;78(3):827-33. doi: 10.1016/j.talanta.2008.12.058. Epub 2009 Jan 15.
A quartz crystal microbalance (QCM) immunosensor was developed for the determination of the insecticide carbaryl and 3,5,6-trichloro-2-pyridinol (TCP), the main metabolite of the insecticide chlorpyrifos and of the herbicide triclopyr. The detection was based on a competitive conjugate-immobilized immunoassay format using monoclonal antibodies (MAbs). Hapten conjugates were covalently immobilized, via thioctic acid self-assembled monolayer (SAM), onto the gold electrode sensitive surface of the quartz crystal. This covalent immobilization allowed the reusability of the modified electrode surface for at least one hundred and fifty assays without significant loss of sensitivity. The piezoimmunosensor showed detection limits (analyte concentrations producing 10% inhibition of the maximum signal) of 11 and 7 microg l(-1) for carbaryl and TCP, respectively. The sensitivity attained (I(50) value) was around 30 microg l(-1) for both compounds. Linear working ranges were 15-53 microg l(-1) for carbaryl and 13-83 microg l(-1) for TCP. Each complete assay cycle took 20 min. The good sensitivity, specificity, and reusability achieved, together with the short response time, allowed the application of this immunosensor to the determination of carbaryl and TCP in fruits and vegetables at European regulatory levels, with high precision and accuracy.
开发了一种石英晶体微天平(QCM)免疫传感器,用于测定杀虫剂西维因以及杀虫剂毒死蜱和除草剂三氯吡氧乙酸的主要代谢产物3,5,6-三氯-2-吡啶醇(TCP)。检测基于使用单克隆抗体(MAb)的竞争性共轭固定免疫分析形式。半抗原共轭物通过硫辛酸自组装单层(SAM)共价固定在石英晶体的金电极敏感表面上。这种共价固定使得修饰电极表面能够重复使用至少150次检测,且灵敏度无显著损失。该压电免疫传感器对西维因和TCP的检测限(产生最大信号10%抑制的分析物浓度)分别为11和7 μg l⁻¹。两种化合物的灵敏度(I(50)值)均约为30 μg l⁻¹。西维因的线性工作范围为15 - 53 μg l⁻¹,TCP的线性工作范围为13 - 83 μg l⁻¹。每个完整的检测周期耗时20分钟。所实现的良好灵敏度、特异性和可重复使用性,以及较短的响应时间,使得该免疫传感器能够高精度、准确地应用于欧洲监管水平下水果和蔬菜中西维因和TCP的测定。