Development of an enzyme-linked immuno-sorbent assay (ELISA) method for carbofuran residues.
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.
Yang
Jinyi
J
College of Food Science, South China Agricultural University, Guangzhou 510642, PR China. yjy361@163.com
Wang
Hong
H
Jiang
Yueming
Y
Sun
Yuanming
Y
Pan
Ke
K
Lei
Hongtao
H
Wu
Qing
Q
Shen
Yudong
Y
Xiao
Zhili
Z
Xu
Zhenlin
Z
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
D000911:Antibodies, Monoclonal; D000941:Antigens; D006241:Haptens; D010573:Pesticide Residues; D012987:Soil; D014867:Water; D006735:Horseradish Peroxidase; D002235:Carbofuran