螺虫乙酯是一种杀虫杀螨剂,它通过抑制乙酰辅酶A羧化酶活性,阻断害虫正常的能量代谢,最终致其死亡。由于其作用高效,被广泛应用于防治果蔬中的刺吸口器类害虫和害螨。然而,螺虫乙酯对人和动物有毒性。该研究旨在通过制备螺虫乙酯的单克隆抗体并建立酶联免疫分析方法实现蔬菜、水果和环境样品中螺虫乙酯残留的快速检测。首先设计合成了一种新型的螺虫乙酯半抗原,通过活泼酯法偶联牛血清白蛋白得到免疫原,免疫小鼠筛选得到特异性识别螺虫乙酯和B-enol的单克隆抗体(mAb 3D11G7)。在优化的条件下,建立间接竞争酶联免疫分析方法(indirect homologous competitive enzyme-linked immunosorbent assay,icELISA),其半数抑制质量浓度(half maximal inhibitory concentration,IC50)值为2.1 μg/L,最佳工作范围为0.5～8.6 μg/L。选择河水、土壤、番茄和柑橘基质进行加标回收实验,其平均加标回收率为72.9%～110.1%。该研究建立的icELISA与超高效液相色谱-串联质谱测定柑橘样品结果的相关性R2为0.965 2,即两种方法的检测结果具有一致性。以上结果表明,该研究开发的icELISA能够用于监测蔬菜、水果和环境样品中螺虫乙酯的残留。
Spirotetramat, a new type of insecticide which can destroy the energy metabolism of pests by acting as an acetyl-CoA carboxylase inhibitor, is widely used in the prevention of sucking pests on vegetables and fruits. However, spirotetramat can cause eye impairment and skin sensitization for animals and human beings. The aim of this study was to achieve rapid detection of spirotetramat residues in vegetables, fruits and environmental samples by preparing monoclonal antibody of spirotetramat and establish an enzyme-linked immunoassay. The spirotetramat hapten was conjugated to BSA to prepare immunogen by the active ester method. One monoclonal antibody (mAb 3D11G7) against both spirotetramat and its metabolite B-enol was prepared. Under optimized conditions, an indirect homologous competitive enzyme-linked immunosorbent assay (icELISA) method was developed. The inhibitor median concentration (IC50) value was 2.1 μg/L and the working range was 0.5-8.6 μg/L. The recoveries of spirotetramat detected in water from Yangtze River, soil, tomato and citrus by icELISA method were ranged from 72.9% to 110.1%. The concentrations of spirotetramat in citrus samples detected by icELISA was in accordance with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) (R2=0.965 2). All these results showed that the developed icELISA method can be used to monitor spirotetramat residue in vegetables, fruits and environmental samples.
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