title: Development of an enzyme-linked immuno-sorbent assay (ELISA) method for carbofuran residues.
abstract: The haptens 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)carbonyl]-amino]butanoic acid (BFNB) and 6-[((2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)-carbonylamino]hexanoic acid (BFNH) were synthesized and then used to develop a rapid,specific and sensitive ELISA method to determine residues of the pesticide carbofuran in a variety of matrices. A hybridoma cell line (5D3) producing anti-carbofuran monoclonal antibodies (MAbs) was also established. Based on the MAbs in combination with the heterologous hapten BFNH coupled to either horseradish peroxidase (HRP) or ovalbumin(OVA), four ELISAs (formats I-IV) for the quantification of carbofuran were developed and compared. Among them, the optimized format II (the conjugate-coated direct competitive ELISA) showed the best characteristics, with an IC50 value of 18.49 ng/mL, a limit of detection of 0.11 ng/mL and the shortest assay time (1 h). This ELISA method was then applied to the determinations of carbofuran in environmental water, soil and food samples. The relative standard deviations (R.S.D.s) ranged from 1.8% to 21.3% and the mean recoveries were 104.6%, 108.3%, 106.3% and 100.1% for water, soil, lettuce and cabbage, respectively. Thus, the ELISA method of format II exhibited the potential to develop commercial ELISA kits for a rapid detection of carbofuran for human health and environmental safety.
authors.affiliation: College of Food Science, South China Agricultural University, Guangzhou 510642, PR China. yjy361@163.com
authors.forename: Jinyi
authors.initials: J
authors.lastname: Yang
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Yueming
authors.initials: Y
authors.lastname: Jiang
authors.forename: Yuanming
authors.initials: Y
authors.lastname: Sun
authors.forename: Ke
authors.initials: K
authors.lastname: Pan
authors.forename: Hongtao
authors.initials: H
authors.lastname: Lei
authors.forename: Qing
authors.initials: Q
authors.lastname: Wu
authors.forename: Yudong
authors.initials: Y
authors.lastname: Shen
authors.forename: Zhili
authors.initials: Z
authors.lastname: Xiao
authors.forename: Zhenlin
authors.initials: Z
authors.lastname: Xu
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification; D000941:Antigens / Q000737:chemistry; D001937:Brassica / Q000737:chemistry; D002235:Carbofuran / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006241:Haptens / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000378:metabolism; D018545:Lactuca / Q000737:chemistry; D010573:Pesticide Residues / Q000032:analysis*; D012987:Soil / Q000032:analysis; D014867:Water / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D006241:Haptens; D010573:Pesticide Residues; D012987:Soil; D014867:Water; D006735:Horseradish Peroxidase; D002235:Carbofuran