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Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Difenoconazole Residues in Fruits and Vegetables

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Abstract

An indirect, competitive enzyme-linked immunosorbent assay (ic-ELISA) for the detection of difenoconazole was developed. Two haptens were designed and successfully synthesized. Hapten 1 had a particular moiety of difenoconazole, while hapten 2 had its full structure. The polyclonal antibodies against hapten-protein conjugates were prepared by immunizing rabbits. After optimization of the conditions, the detection limit (IC15) and sensitivity (IC50) were 4.58 and 29.10 μg L−1, respectively. The cross-reactivities of the antibody with 11 triazole fungicides were all less than 0.1%, which showed that the antibody had excellent specificity. The recoveries of difenoconazole from the spiked samples ranged from 89.70 to 102.31% with good accuracy. The matrix effect was easily removed using a simple, rapid, and efficient extraction method on fruits and vegetables. The detection limit was all 229 μg kg−1 in fruits and vegetables. To validate the ic-ELISA, samples were spiked with difenoconazole at three different concentrations and simultaneously analyzed using high-performance liquid chromatography (HPLC). The results showed a good correlation between the ic-ELISA and HPLC data (R 2 = 0.9970). As a result, the developed immunosorbent assay is suitable for the quantitative determination of difenoconazole in fruits and vegetables.

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Authors and Affiliations

  1. Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, Tianjin Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin, 300457, China

    Bing Liu, Jiuhui Feng, Xiao Sun, Wei Sheng, Yan Zhang & Shuo Wang

Authors
  1. Bing Liu
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  2. Jiuhui Feng
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  3. Xiao Sun
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  4. Wei Sheng
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  5. Yan Zhang
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  6. Shuo Wang
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Corresponding authors

Correspondence to Bing Liu or Shuo Wang.

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Funding

This study was funded by the National Natural Science Foundation of China (Project No. 31301462) and the Ministry of Science and International Science and Technology Cooperation Program of China (Project No. 2014DFR30350).

Conflict of Interest

Bing Liu declares that she has no conflict of interest. Jiuhui Feng declares that she has no conflict of interest. Xiao Sun declares that she has no conflict of interest. Wei Sheng declares that she has no conflict of interest. Yan Zhang declares that she has no conflict of interest. Shuo Wang declares that he has no conflict of interest.

Ethical Approval

All applicable international, national, and institutional guidelines for the care and use of laboratory animals were followed. This article does not contain any studies with human participants performed by any of the authors.

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Liu, B., Feng, J., Sun, X. et al. Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Difenoconazole Residues in Fruits and Vegetables. Food Anal. Methods 11, 119–127 (2018). https://doi.org/10.1007/s12161-017-0983-2

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  • DOI: https://doi.org/10.1007/s12161-017-0983-2

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