Optimization and Validation of an Indirect Competitive ELISA for Detecting Enrofloxacin Residue

Abstract:

This article aimed to optimize a polyclonal antibody based indirect competitive ELISA (icELISA) and compare between the icELSIA method and the LC-MS technology for the determination of enrofloxacin (ENR) residue. Based on the square matrix titration, linear range of the icELISA was from 0.006 to 31.5 ng/mL, with LOD and IC50 values of 0.003 ng/mL and 0.45 ng/mL, respectively. After optimization, 0.03 mol/L of HCl was used in the assay buffer and this ELISA system can tolerate acetonitrile not higher than 10%. The recoveries of ENR were in the range of 98.3-127.5%, 85.7-112.5% and 97.4-103.8% for chicken muscle, liver and kidney. The correlation coefficients (R2) of the ELISA and LC-MS data were 0.9472 in muscle, 0.9843 in liver, and 0.9382 in kidney, respectively.

• Periodical: Advanced Materials Research (Volumes 424-425)
• Pages: 603-606
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.424-425.603
• Online since: January 2012
• Authors: Shi Xiu Wu, Jin Qing Jiang
• Keywords: Chicken, Enrofloxacin, Indirect Competitive ELISA, Optimization

References

[1] G. Yang, B. Lin, Z. Zeng, et al. in: Multiresidue determination of eleven quinolones in milk by liquid chromatography with fluorescence detection [J]. J AOAC Int, 2005, 88(6): 1688-1694.

DOI: 10.1093/jaoac/88.6.1688

[2] H. Yan, H. Wang, X. Qin, et al. in: Ultrasound-assisted dispersive liquid–liquid microextraction for determination of fluoroquinolones in pharmaceutical wastewater [J]. J Pharm Biomed Anal., 2011, 54(1): 53-57.

DOI: 10.1016/j.jpba.2010.08.007

[3] S. Zhao, H. Jiang, X. Li, et al. in: Simultaneous determination of trace levels of 10 quinolones in swine, chicken, and shrimp muscle tissues using HPLC with programmable fluorescence detection [J]. J Agric Food Chem, 2007, 55(10): 3829-3834.

DOI: 10.1021/jf0635309

[4] B. Delepine, D. Hurtaud-Pessel and P. Sanders, in: Simultaneous determination of six quinolones in pig muscle by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [J]. Analyst, 1998, 123(12): 2743-2747.

DOI: 10.1039/a804911d

[5] G. Dufresne, A. Fouquet and D. Forsyth, in: Multiresidue determination of quinolone and fluoroquinolone antibiotics in fish and shrimp by liquid chromatography/tandem mass spectrometry [J]. J AOAC Int, 2007, 90(2): 604-612.

DOI: 10.1093/jaoac/90.2.604

[6] J.Q. Jiang, H.T. Zhang, G.L. Li, et al. in: Preparation of anti-nortestosterone antibodies and development of an indirect heterologous competitive enzyme-Linked immunosorbent assay to detect nortestosterone residues in animal urine [J]. Analytical letters, 2011, 44(14): 2373-2393.

DOI: 10.1080/00032719.2010.551694