Development of an enzyme-linked immunosorbent assay for a broad spectrum triazole fungicide: Hexaconazole

Polyclonal antibodies were produced against 5-(2,4-dichlorophenyl)-5-hydroxy-6-(1H-1,2,4-triazol-1-yl)hexanoic acid (hexaconazole-hapten) conjugated to bovine serum albumin. Using these antibodies, an enzyme-linked immunosorbent assay was developed for detecting and quantifying hexaconazole and its metabolites. This assay was sensitive and had a linear range from 1 to 60 ng/mL. The IC50 was 8 ng/mL and the detection limit 0.1 ng/mL. The polyclonal antibodies recognized hexaconazole and its acid and alcohol metabolites. They cross-reacted with penconazole (46%) and propiconazole (43%), but not with diclobutrazol, imazalil, paclobutrazol, or triadimefon. Using this assay, the concentration of hexaconazole in a commercial fungicide was determined.