Quantitative Determination of Diuron in Ground and Surface Water by Time-Resolved Fluoroimmunoassay: Seasonal Variations of Diuron, Carbofuran, and Paraquat in an Agricultural Area

The aim of this research is to develop an ultrasensitive time-resolved fluorescence immunoassay (TR-FIA) for herbicide diuron in water samples. This method appears to be a promising approach, instead of conventional analytical techniques, in the screening procedure of organic pollutants because it is simple, rapid, and specific, and it does not require sample preconcentration or cleanup. Lanthanide chelate used as label allows to achieve sensitivity even 10 times higher than most of the other techniques. It has been applied to monitoring diuron contamination in specimens collected along a year in an agricultural area. The water specimens were collected monthly from lake, well, and irrigation ditch in the agricultural area south of Milan. Assay was performed using diuron-specific polyclonal antibody raised in sheep; as fluorescent marker, we used rabbit antisheep IgG conjugated with a chelating molecule complexed with Eu3+. The compound 4-(3-(3,4-dichloro-phenyl)-1-methyl-ureido)-butyric acid (CPD) was synthesized and conjugated with bovine serum albumin (BSA) to prepare a solid phase. Sensitivity achieved was 20 ng L-1 below the European Community limits. Paraquat (PQ) and carbofuran (CF) presence in the same samples has been also evaluated in a similar way, using immunoassays with time-resolved revelation systems. Diuron concentration shows a peak coinciding with a peak of carbofuran during summer periods. The peak of diuron was 65 pg/mL in June and 180 pg/mL in September in ditch and lake water samples, respectively; carbofuran concentration was higher than diuron in all samples: a carbofuran peak was revealed in September and October resulting in 87 ng/mL. Herbicide paraquat was not detectable in any assayed sample.