Abstract
For simultaneously monitoring two classes of organophosphorus pesticide residues in food samples, a polyclonal antibody against a dual-generic hapten immunogen, which was prepared by conjugating two generic haptens (O,O-dimethyl thiophosphate hapten and O,O-diethyl phosphate hapten) to the same carrier protein BSA, was raised and used to develop a broad-specificity competitive indirect enzyme-linked immunosorbent assay. With the optimized conditions, the developed immunoassay was able to detect 11 organothiophosphate pesticide and 5 organophosphate pesticide residues with detection limits of 0.018–0.135 μg/mL and 0.013–0.171 μg/mL, respectively. Recoveries of paraoxon-ethyl, fenamiphos, parathion-methyl, and fenthion from spiked samples ranged between 85.8 and 105.5 %. The results indicated that the immunoassay developed in this study is suitable for high-throughput screening of these low-molecular-weight contaminants in food samples.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alcocer MJ, Dillon PP, Manning BM, Doyen C, Lee HA, Daly SJ, O'Kennedy R, Morgan MR (2000) Use of phosphonic acid as a generic hapten in the production of broad specificity anti-organophosphate pesticide antibody. J Agric Food Chem 48:2228–2233
Alder L, Greulich K (2008) Fast multiresidue screening of 300 pesticides in water for human consumption by LC−MS/MS. Anal Bioanal Chem 391:183–197
Banks JN, Chaudhry MQ, Matthews WA, Haverly M, Watkins T, Northway BJ (1998) Production and characterization of polyclonal antibodies to the common moiety of some organophosphorous pesticides and development of a generic type ELISA. Food Agric Immunol 10:349–361
Ercegovich CD, Vallejo RP, Gettig RR, Woods L, Bogus ER, Mumma RO (1981) Development of a radioimmunoassay for parathion. J Agric Food Chem 29:559–563
Gui WJ, Liu YH, Wang CM, Liang X, Zhu GN (2009) Development of a direct competitive enzyme-linked immunosorbent assay for parathion residue in food samples. Anal Biochem 393:88–94
Hennion MC, Barcelo D (1998) Strengths and limitations of immunoassay for effective and efficient use for pesticide analysis in water samples: a review. Anal Chim Acta 362:3–34
Hernándze F, Sancho JV, Pozo OJ (2005) Critical review of theapplication of liquid chromatography/mass spectrometry to thedetermination of pesticide residues in biological samples. Anal Bioanal Chem 382:934–946
Jang MS, Lee SJ, Xue X, Kwon HM, Ra CS, Lee YT, Chung T (2002) Production and characterization of monoclonal antibodies to a generic hapten for class-specific determination of organophosphorus pesticides. Bull Korean Chem Soc 23:1116–1120
Jiang W, Wang Z, Beier RC, Jiang H, Wu Y, Shen J (2013) Simultaneous determination of 13 fluoroquinolone and 22 sulfonamide residues in milk by a dual-colorimetric enzyme-linked immunosorbent assay. Anal Chem 85:1995–1999
Johnson JC, Van Emon JM, Pullman DR, Keeper KR (1998) Development and evaluation of antisera for detection of the OO-diethyl phosphorothionate and phohosphorothionothiolate organophosphorus pesticides by immunoassay. J Agric Food Chem 46:3116–3123
Kim YJ, Cho YA, Lee HS, Lee YT, Gee SJ, Hammock BD (2003) Synthesis of haptens for immunoassay of organophosphorus pesticides and effect of heterology in hapten spacer arm length on immunoassay sensitivity. Anal Chim Acta 475:85–96
Kolosova AY, Park JH, Eremin SA, Kang SJ, Chung DH (2003) Fluorescence polarization immunoassay based on amonoclonal antibody for the detection of the organophosphorus pesticide parathion-methyl. J Agric Food Chem 51:1107–1114
Li J, Liu J, Zhang HC, Li H, Wang JP (2010) Broad specificity indirect competitive immunoassay for determination of nitrofurans in animal feeds. Anal Chim Acta 678:1–6
Liang Y, Liu XJ, Liu Y, Yu XY, Fan MT (2008) Synthesis of three haptens for the class-specific immunoassay of O, O-dimethyl organophosphorus pesticides and effect of hapten heterology on immunoassay sensitivity. Anal Chim Acta 615:174–183
Liu Y, Lou Y, Xu D, Qian G, Zhang Q, Wu R, Hu B, Liu F (2009) Production and characterization of monoclonal antibody for class-specificdetermination of OO-dimethyl organophosphorus pesticides and effect of heterologous coating antigens on immunoassay sensitivity. Microchem J 93:36–42
Lu HT, Le JC, Cheng CS (2005) Measurement method for the hapten coupling ratio of clenbuterol hydrochloride. China Feed 7:33–35
Lu C, Barr DB, Pearson MA, Waller LA (2008) Dietary intake and its contribution to longitudinal organophosphorus pesticide exposure in urban/suburban children. Environ Health Perspect 116:537–542
Mitchell BL, Billingsley BG, Logue BA (2013) Rapid quantification of dimethyl methylphosphonate from activated carbon particles by static headspace gas chromatography mass spectrometry. J Chromatogr A1293:120–125
Piao YZ, Kim YJ, Kim YA, Lee HS, Hammock BD, Lee YT (2009) Development of ELISAs for the class-specific determination of organophosphorus pesticides. J Agric Food Chem 57:10004–10013
Pinacho DG, Sánchez-Baeza F, Marco M (2012) Molecular modeling assisted hapten design to produce broad selectivity antibodies for fluoroquinolone antibiotics. Anal Chem 84:4527–4534
Wang ST, Gui WJ, Guo YR, Zhu GN (2007a) Preparation of a multi-hapten antigen and broad specificity polyclonal antibodies for a multiple pesticide immunoassay. Anal Chim Acta 587:287–292
Wang Z, Zhu Y, Ding S, He F, Beier RC, Li J, Jiang H, Feng C, Wan Y, Zhang S, Kai Z, Yang X, Shen J (2007b) Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds. Anal Chem 79:4471–4483
Xu ZL, Shen YD, Zheng WX, Beier RC, Xie GM, Dong JX, Yang JY, Wang H, Lei HT, She ZG, Sun YM (2010) Broad-specificity immunoassay for O, O-diethyl organophosphorus pesticides: application of molecular modeling to improve assay sensitivity and study antibody recognition. Anal Chem 82:9314–9321
Yang ZY, Kolosova AY, Shim WB, Chung DH (2006) Development of monoclonal antibodies against pirimiphos-methyl and their application to IC-ELISA. J Agric Food Chem 54:4551–4556
Yuan M, Liu B, Liu EM, Sheng W, Zhang Y, Crossan A, Kennedy I, Wang S (2011) Immunoassay for phenylurea herbicides: application of molecular modeling and quantitative structure-activity relationship analysis on an antigen-antibody interaction study. Anal Chem 83:4767–4774
Zhao Q, Lu Q, Yu QW, Feng YQ (2013) Dispersive microextraction based on “magnetic water” coupled to gas chromatography/mass spectrometry for the fast determination of organophosphorus pesticides in cold-pressed vegetable oils. J Agric Food Chem 61:5397–5403
Acknowledgments
This study was supported by the National Natural Science Foundation of China (grant numbers 30972050 and 31271873), the Natural Science Foundation of Shandong province (grant number ZR2010CM015), the Genetically Modified Organisms Breeding Major Projects (grant number 2009ZX08012-003B), and the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry.
Conflict of Interest
Yanling Li declares that she has no conflict of interest. Fengchun Zhao declares that he has no conflict of interest. Longyu Zhao declares that she has no conflict of interest. Zhengyou Yang declares that he has no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, Y., Zhao, F., Zhao, L. et al. Development of a Broad-Specificity Immunoassay for Determination of Organophosphorus Pesticides Using Dual-Generic Hapten Antigens. Food Anal. Methods 8, 420–427 (2015). https://doi.org/10.1007/s12161-014-9906-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-014-9906-7