CONFERENCE PROCEEDING
New tracers for fluorescence polarization immunoassay of herbicide 4-chlorophenoxyacetic acid
1
Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia
2
A.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Moscow, Russia
Publication date: 2026-07-30
Public Health Toxicol 2026;6(Supplement 1):A10
KEYWORDS
ABSTRACT
Introduction:
Chlorophenoxy herbicides are a widely used class of agricultural compounds employed for weed control and as plant growth regulators. Among these, 4-chlorophenoxyacetic acid (4-D) is commonly utilized as both a herbicide and a growth regulator. However, herbicides are often applied at higher concentrations than necessary, increasing the risk of contamination in food products, which may pose hazards to human health and the environment. Therefore, developing a method to detect residual traces of 4-D is highly relevant. Fluorescence polarization immunoassay (FPIA) presents a promising solution for this purpose. The aim of this study was to synthesize tracers for 4-D detection and optimize FPIA conditions.
Methods:
Two tracers of 4-chlorophenoxyacetic acid were synthesized using the carbodiimide method, conjugating the compound with fluorescein derivatives: ethylene-diamine-fluorescein thiocarbamate (EDF) and 5-aminohexyl-amino-carbonyl-fluorescein (5AHCF). The tracers' structures were confirmed by mass spectrometry. Monoclonal antibodies (M1 and M2) against 4-D were sourced from China.
Results:
Both tracers yielded comparable results. However, further work proceeded with the 4D-EDF tracer and M1 monoclonal antibody. The FPIA for 4-D was optimized by determining the ideal concentrations of the tracer and M1 antibody, achieving a bound fraction of 0.5. The optimal concentrations were 2.5 nM for the tracer and 4 nM for the antibody. The assay demonstrated a detection limit of 1.2 ng/mL in aqueous solutions, with an analysis time of 15 minutes. Accuracy was confirmed via a spike-and-recovery test, yielding recovery rates between 95% and 115%. Parallel ELISA validation showed a relative standard deviation of 3%, confirming the reliability of 4-D detection in real water samples.
Conclusion:
The 4D-EDF tracer is suitable for detecting 4-D in water. FPIA offers several advantages, including rapid analysis, high sensitivity, and minimal sample preparation, making it a practical tool for environmental monitoring.
Chlorophenoxy herbicides are a widely used class of agricultural compounds employed for weed control and as plant growth regulators. Among these, 4-chlorophenoxyacetic acid (4-D) is commonly utilized as both a herbicide and a growth regulator. However, herbicides are often applied at higher concentrations than necessary, increasing the risk of contamination in food products, which may pose hazards to human health and the environment. Therefore, developing a method to detect residual traces of 4-D is highly relevant. Fluorescence polarization immunoassay (FPIA) presents a promising solution for this purpose. The aim of this study was to synthesize tracers for 4-D detection and optimize FPIA conditions.
Methods:
Two tracers of 4-chlorophenoxyacetic acid were synthesized using the carbodiimide method, conjugating the compound with fluorescein derivatives: ethylene-diamine-fluorescein thiocarbamate (EDF) and 5-aminohexyl-amino-carbonyl-fluorescein (5AHCF). The tracers' structures were confirmed by mass spectrometry. Monoclonal antibodies (M1 and M2) against 4-D were sourced from China.
Results:
Both tracers yielded comparable results. However, further work proceeded with the 4D-EDF tracer and M1 monoclonal antibody. The FPIA for 4-D was optimized by determining the ideal concentrations of the tracer and M1 antibody, achieving a bound fraction of 0.5. The optimal concentrations were 2.5 nM for the tracer and 4 nM for the antibody. The assay demonstrated a detection limit of 1.2 ng/mL in aqueous solutions, with an analysis time of 15 minutes. Accuracy was confirmed via a spike-and-recovery test, yielding recovery rates between 95% and 115%. Parallel ELISA validation showed a relative standard deviation of 3%, confirming the reliability of 4-D detection in real water samples.
Conclusion:
The 4D-EDF tracer is suitable for detecting 4-D in water. FPIA offers several advantages, including rapid analysis, high sensitivity, and minimal sample preparation, making it a practical tool for environmental monitoring.
CONFLICTS OF INTEREST
The authors declare that they have no conflicts of interest in the publication of this article. The authors have no conflicts of interest to report in this work. Abstract was not submitted elsewhere and published here firstly.
FUNDING
The study was supported by the Russian Science Foundation; grant # 24-43-00196 (https://rscf.ru/project/24-43-00196/).
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| eISSN: | 2732-8929 |
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