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Detection of Di-Butyl Phthalate in Surface Water by Fluorescence Polarization Assay and Gas Chromatography -Mass Spectrometry
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This study developed a fast fluorescence-based immunoassay for detecting dibutyl phthalate (DBP) — a common plasticizer with known reproductive and developmental toxicity — in surface water. The method is 35 times more sensitive than previous approaches and was successfully validated on real water samples, offering a practical tool for monitoring this plastic-derived contaminant in the environment.
Dibutyl phthalate (DBP) is widely used as a plasticizer in the production of polymeric materials to give them flexibility, strength and extensibility. However, due to its negative impact on human health, in particular, reproductive function and fetal development, the content of DBP must be controlled in food and environmental objects. The present study aims to develop a sensitive, fast and simple polarization fluorescent immunoassay (FPIA) using monoclonal antibodies derived against DBP (MAb-DBP) for its detection in open waters. New conjugates of DBP with various fluorescein derivatives were obtained and characterized: 5-aminomethylfluorescein (AMF) and dichlorotriazinylaminofluorescein (DTAF). The advantages of using the DBP-AMF conjugate in the FPIA method are shown, the kinetics of binding of this drug with antibodies is studied, the analysis is optimized, and the concentration of monoclonal antibodies is selected for sensitive analysis - 16 nM. The calibration dependence of the fluorescence polarization signal for the detection of DBP was obtained. The observed IC50 (40 ng/mL) and LOD (10 ng/mL) values were improved by a factor of 35 for the previously described FPIA using polyclonal antibodies. This technique was tested by the added/found method, the percentage of DBP discovery in water ranged from 85 to 110%. Using the developed method, real water samples from Lake Onega were tested, and a good correlation was shown between the results of the determination of DBP by the FPIA method and GC-MS/MS. Thus, the FPIA method developed in this work can be used to determine DBP in open water reservoirs.
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Researchers developed a fluorescence polarization immunoassay for detecting dibutyl phthalate, a common plasticizer, in surface water samples. The study demonstrates that this method provides a rapid and sensitive way to monitor phthalate contamination in water, which is relevant given concerns about the health effects of plasticizer exposure.
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