Application of a monoclonal-based immunoassay for the determination of imazalil in fruit juices

A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of imazalil (RS)-1-(β-allyloxy-2,4-dichlorophenylethyl)imidazole] in apple, tomato and orange juice samples. From an imazalil hapten, which mimics the analyte structure, several monoclonal antibodies were obtained. An ELISA in the conjugate-coated format was developed and optimized using the antibody showing the highest sensitivity. For standards, the detection limit of the ELISA was 0.2?nM (0.06?ng?ml^-1), with an I₅₀ value of 1.6?nM (0.5?ng?ml^-1). The study of the influence of matrices on assay reliability indicated that the ELISA could determine imazalil in fruit juices at the low ng?ml^-1 level simply by diluting the sample, without any clean-up or concentration step. Recovery and precision of the method were evaluated by spiking juice samples with imazalil in the 10-500?ng?ml^-1 range. The mean recovery from fruit juices was 97% and the mean coefficient of variation was ～20%. In addition to being precise and accurate, the method has proved to be simple and sensitive, with a quantification limit well below the maximum residue limits for imazalil in these matrices.