Manclús Juan J, Moreno María J, Plana Emma, Montoya Angel
Instituto de Investigación e Innovación en Bioingeniería, Universidad Politécnica de Valencia, 46022 Valencia, Spain.
J Agric Food Chem. 2008 Oct 8;56(19):8793-800. doi: 10.1021/jf801187b. Epub 2008 Sep 11.
Enzyme-linked immunosorbent assays (ELISAs) based on monoclonal antibodies for the detection of triazole fungicides have been developed. With this aim, hapten-protein conjugates, containing the common triazole and chlorinated aromatic moieties, were prepared. From mice immunized with these conjugates, several monoclonal antibodies (MAbs) with the ability to sensitively bind several triazoles with different specificity were obtained. Both analyte- and class-specific ELISAs were developed. The hexaconazole-specific immunoassay can determine this fungicide with a limit of detection of 0.3 mug/L in standard buffer. The so-called triazole-specific immunoassay allowed for the detection of tetraconazole, penconazole, cyproconazole, and myclobutanil, with limits of detection in the 0.1-0.7 mug/L range. These immunoassays were applied to the determination of triazoles in spiked fruit juices. Samples were adequately diluted to minimize the matrix effects. Coefficients of variation were below 30%, and recoveries ranged from 62 to 135%. Therefore, the developed immunoassays can determine triazole fungicides in fruit juices down to the maximum residue limits currently legislated, without any sample treatment other than dilution.
已开发出基于单克隆抗体的酶联免疫吸附测定法（ELISA）用于检测三唑类杀菌剂。为此，制备了含有常见三唑和氯化芳香部分的半抗原-蛋白质缀合物。用这些缀合物免疫小鼠后，获得了几种能够以不同特异性灵敏结合多种三唑的单克隆抗体（MAb）。同时开发了分析物特异性和类别特异性的ELISA。己唑醇特异性免疫测定法在标准缓冲液中检测该杀菌剂的检测限为0.3μg/L。所谓的三唑类特异性免疫测定法可检测戊唑醇、戊菌唑、环丙唑醇和腈菌唑，检测限在0.1-0.7μg/L范围内。这些免疫测定法用于测定加标果汁中的三唑类。将样品充分稀释以尽量减少基质效应。变异系数低于30%，回收率在62%至135%之间。因此，所开发的免疫测定法无需除稀释以外的任何样品处理，即可测定果汁中低至目前法定最大残留限量的三唑类杀菌剂。
