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A sensitive and group-specific monoclonal antibody-based indirect competitive ELISA for the determination of salbutamol in swine meat and liver samples

Abstract

Salbutamol (SAL), a kind of β-adrenergic receptor agonists (β-agonists) which can enhance the lean meat-to-fat ratio, has been inhibited as an additive used in animal feeds for livestock production in many countries due to its harmful effect to the consumers. Herein we presented a sensitive and group-specific ELISA based on monoclonal antibody (mAb) for the determination of SAL in swine meat and liver samples. The SAL derivative containing carboxylic group was prepared by directly reacting SAL with succinic anhydride, then the derivative was linked to carrier proteins. Mice immunized with SAL-bovine serum albumin (BSA) conjugate were utilized for mAb generation. The hybridoma clone (1C3) specifically secreting mAb against SAL with the higher sensitivity was successfully obtained. Under optimal conditions, the IC50 and LOD values of the mAb-based ELISA for SAL were found to be 0.47 ng mL-1 and 0.049 ng mL-1, respectively, indicating high sensitivity of the assay. The mAb displayed high cross-reactivity (CR) with four β-agonists, e.g. clenbuterol (55%), terbutaline (44%), brombuterol (33%) and cimbuterol (134%) because these compounds contain the same tert-butyl group on the nitrogen atom like SAL has. The CR of the mAb with other β-agonists including cimaterol, clorprenaline, isoprenaline, neophryn, ractopamine and phenylethylamine A was less than 2.2% due to relatively large structure difference comparing to that of SAL. Swine meat and liver samples were spiked with different content of SAL and detected by ELISA. The recovery rates were respectively found in the range of 76.4-107.6% and 71.8-106.8%, with coefficients of variation less than 20%. Spiked samples were also analyzed by HPLC. A good correlation between two methods was obtained. These results demonstrated that the proposed ELISA was a feasible quantitative/screening method for SAL analysis in swine meat and liver samples. In addition, the recoveries results of the β-agonists which displayed the high CR values with mAb in an additional spiking experiment revealed the ELISA could be also as an alternative method for multiple β-agonists detection.

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Supplementary files

Publication details

The article was received on 04 Jul 2017, accepted on 13 Sep 2017 and first published on 14 Sep 2017


Article type: Paper
DOI: 10.1039/C7AY01628J
Citation: Anal. Methods, 2017, Accepted Manuscript
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    A sensitive and group-specific monoclonal antibody-based indirect competitive ELISA for the determination of salbutamol in swine meat and liver samples

    A. Deng, Y. Yuan, Y. Zhao, K. Wu, H. Yang, K. Zhao and J. Li, Anal. Methods, 2017, Accepted Manuscript , DOI: 10.1039/C7AY01628J

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