Immunochemical rapid determination of quinoxyfen, a priority hazardous pollutant

In 2013, quinoxyfen was included in the list of priority hazard pollutants of the European Water Framework Directive due to its toxicity to aquatic organisms. However, few analytical methods for the analysis of this fungicide have been reported and no rapid immunochemical methods have been published so far. In the present study, immunoreagents for quinoxyfen analysis were generated for the first time and an enzyme-linked immunosorbent assay was developed. Two carboxylated derivatives of quinoxyfen were designed on the basis of the minimum energy conformation of the target compound. Active esters of those novel compounds were prepared using N,N'-disuccinimidyl carbonate, and purified for covalent coupling to proteins. Matrix-assisted laser desorption mass spectrometry of the prepared bioconjugates showed optimum hapten-to-protein molar ratios. Moreover, high-affinity antibodies specific of quinoxyfen were raised. As proof of concept, an immunoassay was evaluated using a heterologous conjugate, which afforded sensitivity values in the low nanomolar range. Moreover, excellent recoveries and coefficients of variation were obtained from the analysis of environmental water samples fortified with quinoxyfen. A limit of quantification of 60 μg/L was determined. The prepared bioconjugates and antibodies could be valuable immunoreagents for the development of a variety of rapid immunosensors for quinoxyfen determination in environmental samples.