Development of an Immunoassay for the Residues of the Herbicide Bensulfuron-Methyl

• Published in: Journal of agricultural and food chemistry Vol. 50; no. 7; pp. 1791 - 1803
• Main Authors: Lee, Jae Koo, Ahn, Ki Chang, Park, Oee Sook, Ko, Yong Kwan, Kim, Dae-Whang
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 27.03.2002; Amer Chemical Soc
• Subjects: Agriculture; Agriculture, Multidisciplinary; Agronomy. Soil science and plant productions; Analysis methods; Animals; Antibody Formation; Antibody Specificity; Antigens - immunology; Applied sciences; Biological and medical sciences; Chemistry; Chemistry, Applied; Chromatography, High Pressure Liquid; Detergents; Enzyme-Linked Immunosorbent Assay - methods; Exact sciences and technology; Food industries; Food Science & Technology; Food toxicology; Fundamental and applied biological sciences. Psychology; Haptens - chemistry; Haptens - immunology; Hemocyanins - immunology; Herbicides - analysis; Hydrogen-Ion Concentration; Immune Sera - immunology; Immunoassay - methods; Life Sciences & Biomedicine; Magnetic Resonance Spectroscopy; Osmolar Concentration; Pesticide Residues - analysis; Physical Sciences; Pollution; Rabbits; Science & Technology; Sensitivity and Specificity; Soil and sediments pollution; Soil and water pollution; Soil science; Solvents; Sulfonylurea Compounds - analysis; Sulfonylurea Compounds - immunology; SULFONYLUREA HERBICIDES; CHLORSULFURON; ENZYME-IMMUNOASSAY; bensulfuron-methyl; LINKED-IMMUNOSORBENT-ASSAY; sulfonylurea herbicide; monitoring; GAS-CHROMATOGRAPHY; LIQUID-CHROMATOGRAPHY; CAPILLARY-ELECTROPHORESIS; ELISA; polyclonal antibodies; S-TRIAZINE HERBICIDES; METSULFURON-METHYL; WATER; Antibody; Pesticides; Herbicide; Hapten; Optimization; Tailings; Pollutant; Toxicology; Sulfonylureas; Detection threshold; Measurement method; Measurement accuracy; ELISA assay
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf011150b

To develop a competitive indirect enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of the sulfonylurea herbicide bensulfuron-methyl, seven structurally related haptens were synthesized. Four of them mimicking the target analyte were conjugated to keyhole limpet hemocyanin by the N-hydroxysuccinimide activated ester method to use as immunogens, and all of them were conjugated to bovine serum albumin to use as plate-coating antigens. Polyclonal antibodies raised in rabbits and the coating antigens were screened and selected for the assay in simple homologous and heterologous ELISA formats. Three sensitive heterologous ELISAs were selected and optimized, showing the average IC50 values of bensulfuron-methyl as low as 0.17, 0.09, and 0.09 ng/mL, the detection ranges of 0.04−0.60, 0.01−0.60, and 0.04−0.25 ng/mL, and the lowest detection limits of 0.03, 0.002, and 0.03 ng/mL, respectively. The cross-reactivities of other sulfonylurea herbicides and metabolites of bensulfuron-methyl to the antibodies were less than 15% in the two assays. Recoveries from the analyte-fortified water samples in assay I were in the range of 81−125% by simple dilution. The correlation between the ELISA and HPLC was 0.999 (n = 15) with a slope of 1.37 in the analysis of groundwater samples fortified with bensulfuron-methyl. The results obtained strongly indicate that the ELISA can be a highly sensitive and convenient tool for detecting bensulfuron-methyl residues in agricultural and environmental samples. Keywords: Bensulfuron-methyl; sulfonylurea herbicide; ELISA; polyclonal antibodies; monitoring