Sheng En Ze, Tan Yu Ting, Lu Yu Xiao, Xiao Yue, Li Zhen Xi
Jiangsu Collaborative Innovation Center of Biomedical Functional Materials and Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China.
Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
Front Chem. 2021 Jan 7;8:621925. doi: 10.3389/fchem.2020.621925. eCollection 2020.
The direct and indirect competition time-resolved fluorescence immunoassays (dc-TRFIA, ic-TRFIA) were established by combining the autofluorescence properties of lanthanide europium (Eu) with the monoclonal antibody of oxyfluorfen. The purified Eu antibody was optimized and the conditions such as the working concentration of the Eu antibody, monoclonal antibody, and working buffer were optimized. In the optimal condition, the IC of dc-TRFIA was 10.27 ng/mL, the lowest detection limit IC was 0.071 ng/mL, the detection range (IC-IC) was 0.071-1074.3 ng/mL, and the detection range (IC-IC) and IC of ic-TRFIA were 0.024-504.6 and 2.76 ng/mL, respectively. The comparison showed that the sensitivity and detection limit of ic-TRFIA were superior to dc-TRFIA. The cross reaction (CR) tests showed that the CR with other oxyfluorfen structure analogs was <0.02%, except that there was a certain CR with the benzofluorfen (CR = 11.58) and the bifenox (CR = 8.23%). The average recoveries of ic-TRFIA were 74.6-108.3%, and the RSDs were between 2.1 and 10.9%, in the addition recovery test with five substrates. The results of the correlation test with the real samples of GC-ECD showed that they were highly correlated ( = 0.975x - 0.4446, = 0.9901), which proved that the TRFIA method established in this study had high reliability and accuracy and could be used in environment and agricultural products for rapid detection of oxyfluorfen residues.
通过将镧系元素铕(Eu)的自身荧光特性与乙氧氟草醚单克隆抗体相结合,建立了直接竞争和间接竞争时间分辨荧光免疫分析法(dc - TRFIA、ic - TRFIA)。对纯化后的Eu抗体进行了优化,并对Eu抗体、单克隆抗体的工作浓度以及工作缓冲液等条件进行了优化。在最佳条件下,dc - TRFIA的IC为10.27 ng/mL,最低检测限IC为0.071 ng/mL,检测范围(IC - IC)为0.071 - 1074.3 ng/mL,ic - TRFIA 的检测范围(IC - IC)和IC分别为0.024 - 504.6和2.76 ng/mL。比较结果表明,ic - TRFIA的灵敏度和检测限优于dc - TRFIA。交叉反应(CR)试验表明,除了与乙羧氟草醚(CR = 11.58)和除草醚(CR = 8.23%)有一定交叉反应外,与其他乙氧氟草醚结构类似物的交叉反应均<0.02%。在对5种底物的加标回收试验中,ic - TRFIA的平均回收率为74.6 - 108.3%,相对标准偏差在2.1%至10.9%之间。与GC - ECD实际样品的相关性测试结果表明,二者具有高度相关性( = 0.975x - 0.4446, = 0.9901),这证明本研究建立的TRFIA方法具有高可靠性和准确性,可用于环境和农产品中乙氧氟草醚残留的快速检测。