Development of an enzyme-linked immunosorbent assay for screening contamination by chlorophenols in environmental waters

Abstract

The development of an immunoassay for screening contamination by chlorophenols is presented. Two haptens were synthesized and conjugated to immunizing proteins to raise rabbit polyclonal antibodies. The antibody-coated format (direct) gave better sensitivity than the conjugate-coated format (indirect) if 2,4,6-trichlorophenol is used as target analyte. The measurement range was 86.4 microg l(-1) to 0.7 microg l(-1), with an average I50 of 7.8 microg l(-1) and a detection limit of 0.2 microg l(-1). The assay detects the presence of trichloropyridinol and other chlorophenols such as di-, tetra- and pentachlorophenols constituting thus a suitable tool for the early warning of the presence of such family contaminants. The optimized method permits the detection of the most important chlorophenols in a fast and reproducible way using no more than one antibody and a single assay. The results achieved with water samples spiked with different chlorophenols fit with a multiple linear regression model when expressing the total concentration of chlorophenols as equivalent of 2,4,6-trichlorophenol (P < 0.01), demonstrating the usefulness of the assay as a screening tool to detect contamination by chlorophenols.