噻虫胺残留时间分辨荧光免疫分析研究..docVIP

噻虫胺残留时间分辨荧光免疫分析研究 李明，盛恩泽，袁玉龙，刘晓凤，施海燕，华修德，王鸣华* （南京农业大学植物保护学院农药科学系，江苏 南京210095） 摘要：本研究以铕离子（Eu3+）作为荧光标记物对羊抗兔IgG进行标记，建立了一种基于多克隆抗体高灵敏检测农业样品中噻虫胺残留的间接竞争时间分辨荧光免疫分析方法（TRFIA）。在最佳优化条件下，建立了噻虫胺的TRFIA标准曲线，抑制中浓度（IC50）为2.07μg/L，最低检测浓度（LOD，IC10）为0.0220 μg/L。建立的TRFIA除了对呋虫胺有较大交叉反应外（9.7%），与噻虫胺结构类似化合物没有明显的交叉反应。水、甘蓝和大米中的平均添加回收率为74.1-115.9%，相对标准偏差为3.7-11.7%。真实样品的检测结果表明TRFIA与气相色谱法具有高度的相关性（R2 = 0.9902）。研究说明建立的TRFIA能够高灵敏地检测农产品中噻虫胺的残留。 关键词：噻虫胺；铕；多克隆抗体；时间分辨荧光免疫分析方法 Sensitive time-resolved fluoroimmunoassay for quantitative determination clothianidin LI Ming, SHENG Enze, YUAN long, LIU Xiaofeng, SHI Haiyan, HUA Xiude, WANG Minghua* (Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China) Abstract: Europium (Eu3+)-labeled antibody was used as a fluorescent label to develop a highly sensitive indirect competitive time-resolved fluoroimmunoassay (TRFIA) for determination of clothianidin residues based on polyclonal antibody in agricultural samples. Under optimal conditions, the half-maximal inhibition concentration (IC50) and the limit of detection (LOD, IC10) of clothianidin were 2.07 and 0.0220 μg/L for the TRFIA, respectively. The TRFIA was no obvious cross-reactivities with the analogues of clothianidin except for dinotefuran (9.7%). Analysis of clothianidin-fortified samples (water, cabbage and rice) by the TRFIA showed average recoveries from 74.1-115.9% with relative standard deviations of 3.7-11.7%. The results of TRFIA for the authentic samples were largely consistent with gas chromatography (R2 = 0.9902). The optimised TRFIA might become a sensitive and satisfactory analytical method for the quantitative monitoring of clothianidin residues in agricultural produces. Keywords: clothianidin; europium; plyclonal antibody; time-resolved fluoroimmunoassay 噻虫胺（clothianidin）是含噻唑环的新型烟碱类杀虫剂，以其新颖的作用机制、广谱高效的杀虫活性、灵活的使用方法，被广泛应用于防治危害水稻、蔬菜、果园、茶叶等作物的半翅目、鞘翅目、双翅目和某些鳞翅目害虫[1]。随着人们对环境和食品安全关注程度增强[2]，对噻虫胺在农产品和环境中残留检测方法的研究也广泛开展。我国尚未制订噻虫胺的允许残留量标准，