Mukhametova Liliya I, Kolokolova Marya K, Shevchenko Ivan A, Tupertsev Boris S, Zherdev Anatoly V, Xu Chuanlai, Eremin Sergei A
Faculty of Chemistry, M.V. Lomonosov Moscow State University, Leninsky Gory 1/3, 119991 Moscow, Russia.
A. N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences; Leninsky Prospect 33, 119071 Moscow, Russia.
Biosensors (Basel). 2025 Jan 9;15(1):32. doi: 10.3390/bios15010032.
2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the popular herbicides that is widely used in agriculture and can be found in food and water. A rapid and sensitive fluorescence polarization immunoassay (FPIA) was proposed for the detection of 2,4-D in juice and water. New tracers, 2,4-D-buthylenediamin fluoresceinthiocarbamyl (2,4-D-BDF) and 2,4-D-glycine aminofluorescein (2,4-D-GAF), were obtained and characterized. Monoclonal antibodies (MAb) obtained against 2,4-D were used as a recognition reagent. The kinetics of the interaction of MAb and tracers were studied, and the kinetic parameters of their binding were calculated. High specificity of binding of tracers and MAb was shown. In this work, an approach was elaborated on to reduce the detection limit of 2,4-D by the FPIA method by changing the volume of the studied sample. The optimized FPIA in a competitive format was characterized by the LODs of 2,4-D 8 and 0.4 ng/mL and the working ranges 30-3000 ng/mL and 3-300 ng/mL for juice and water, respectively. The entire test cycle (from sample receipt to evaluation of the analysis results) took only 20 min. The test for the recovery of 2,4-D in juice and water gave values from 95 to 120%, which demonstrated the reliability of the herbicide determination in real samples.
2,4-二氯苯氧乙酸（2,4-D）是一种广泛应用于农业的常见除草剂，可在食品和水中检测到。本文提出了一种快速灵敏的荧光偏振免疫分析法（FPIA）用于检测果汁和水中的2,4-D。制备并表征了新型示踪剂2,4-二氯苯氧乙酸-丁二胺荧光硫代氨基甲酰（2,4-D-BDF）和2,4-二氯苯氧乙酸-甘氨酸氨基荧光素（2,4-D-GAF）。以抗2,4-D的单克隆抗体（MAb）作为识别试剂。研究了MAb与示踪剂相互作用的动力学，并计算了它们结合的动力学参数。结果表明示踪剂与MAb的结合具有高特异性。在本研究中，详细阐述了一种通过改变被测样品体积来降低FPIA法检测2,4-D检测限的方法。优化后的竞争型FPIA对果汁和水中2,4-D的检测限分别为8 ng/mL和0.4 ng/mL，工作范围分别为30-3000 ng/mL和3-300 ng/mL。整个测试周期（从样品接收至分析结果评估）仅需20分钟。果汁和水中2,4-D的回收率测试结果为95%至120%，这证明了该除草剂在实际样品中测定的可靠性。
