Immunoreagent generation and competitive assay development for cyprodinil analysis

Cyprodinil is an anilinopyrimidine fungicide applied worldwide for the prevention and treatment of highly destructive plant diseases in a large variety of crops, including cereals, fruits, and vegetables. This paper describes the development of the first reported immunoassays for cyprodinil. Two original haptens have been synthesized and conjugated to different carrier proteins, and polyclonal antibodies have been produced. Moreover, competitive enzyme-linked immunosorbent assays have been developed and characterized for the analysis of this widely used pesticide. The influence of organic solvents and buffer conditions over the assay analytical parameters was studied. The IC(50) values of the optimized immunoassays were 1.6 and 2.8 ng/mL for the direct and indirect formats, respectively. Quantitative recoveries were found using spiked apple and grape juice samples after a simple direct dilution, and a limit of quantification of 20 ng/mL for both fruit matrices was achieved. These immunoreagents could be very valuable for the sensitive, straightforward, and rapid monitoring of cyprodinil residues in foodstuffs.