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A Homogeneous Fluorescence Polarization Immunoassay for the Determination of Cephalexin and Cefadroxil in Milk

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Abstract

A new monoclonal antibody (mAb) against cephalexin (CEX), named 1F3, was produced by immunizing mice with immunogen CEX–keyhole limpet hemocyanin. Two fluorescein-labeled tracers (CEX–fluorescein isothiocyanate (FITC) and cefadroxil (CDX)–FITC) were synthesized and purified by thin-layer chromatography. A homogeneous fluorescence polarization immunoassay (FPIA) based on the new mAb and the selected tracer for the simultaneous determination of CEX and CDX was developed and optimized. The optimized FPIA provided a limit of detection of 1.8 and 2.3 ng ml−1 for CEX and CDX and exhibited cross-reactivities of 100 and 63.16 % for CEX and CDX, respectively, and <1 % for other 11 cephalosporins. Recoveries from fortified milk were ranged from 82 to 125 % with coefficient of variations of 3.2–14.2 %. The results indicated that the homogeneous FPIA we developed was enough sensitive to rapidly monitor CEX and CDX in milk.

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Acknowledgments

This work is supported by the grants from Trans-Century Training Programme Foundation for the Talents by the Ministry of Education (NCET-12-0529), Key Projects in the National Science & Technology Pillar Program during the 12th Five-Year Plan Period (2011BAK10B01, 2012BAK17B00) and Sino-Russian International Scientific and Technological Cooperation (2011DFR30470).

Conflict of Interest

Jing Zhang has no competing financial interests. Zhanhui Wang has no competing financial interests. Tiejun Mi has no competing financial interests. Lequn Wenren has no competing financial interests. Kai Wen has no competing financial interests. Guidelines for the care and use of laboratory animals were followed.

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Authors and Affiliations

  1. College of Veterinary Medicine, China Agricultural University, Beijing, 100093, People’s Republic of China

    Jing Zhang, Zhanhui Wang, Tiejun Mi, Lequn Wenren & Kai Wen

  2. National Reference Laboratories for Veterinary Drug Residue, District Haidian, Beijing, 100193, People’s Republic of China

    Jing Zhang, Zhanhui Wang, Tiejun Mi, Lequn Wenren & Kai Wen

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  1. Jing Zhang
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  2. Zhanhui Wang
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  3. Tiejun Mi
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  5. Kai Wen
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Correspondence to Kai Wen.

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Zhang, J., Wang, Z., Mi, T. et al. A Homogeneous Fluorescence Polarization Immunoassay for the Determination of Cephalexin and Cefadroxil in Milk. Food Anal. Methods 7, 879–886 (2014). https://doi.org/10.1007/s12161-013-9695-4

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  • DOI: https://doi.org/10.1007/s12161-013-9695-4

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