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Issue 11, 2001
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A new electrochemical enzyme-linked immunosorbent assay for the screening of macrolide antibiotic residues in bovine meat

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Abstract

A new sensitive electrochemical enzyme-linked immunosorbent assay (ELISA) for the detection of two macrolides (erythromycin and tylosin) in bovine muscle was developed, using the mouse monoclonal antibodies anti-erythromycin and anti-tylosin. The competitive indirect assay was performed using an erythromycin (or tylosin)–BSA conjugate as a coating molecule; after competition between free and coated analytes for the antibodies, the activity of the horseradish peroxidase-labelled antiglobulins was measured electrochemically using 3,3′,5,5′-tetramethylbenzidine (TMB) as substrate. The detection limit of the assay was 0.4 ng ml−1 for erythromycin and 4.0 ng ml−1 for tylosin, while the sensitivity (25% inhibition concentration) was 1.4 ng ml−1 for erythromycin and 13.0 ng ml−1 for tylosin. The specificity of the assay was assessed by studying the cross-reactivity of various macrolides other than erythromycin and tylosin. The results indicate that the monoclonal antibodies anti-erythromycin and anti-tylosin can readily distinguish the target compound from other macrolides, with the exception of roxithromycin, a semisynthetic macrolide antibiotic derived from erythromycin. Fortified and real samples were analysed by the developed ELISA method and results confirmed by micro-LC-MS-MS using an atmospheric pressure ionisation (API) source and an ionspray (IS) interface. The latter provides unequivocal identification and quantification of the analytes at the level of interest. The ELISA assay showed precision (RSD) values ranging from 6.3 to 11.4% for erythromycin and from 7.5 to 12.6% for tylosin; the accuracy (relative error, RE) ranged from −16.0 to −9.8% and from −9.5 to 8.0% for erythromycin and tylosin, respectively. All results obtained demonstrate that the electrochemical ELISA is a suitable method for a sensitive, simple, rapid and reliable screening of the two macrolides in animal tissues.

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Publication details

The article was received on 05 Jun 2001, accepted on 09 Sep 2001 and first published on 22 Oct 2001


Article type: Paper
DOI: 10.1039/B104939A
Citation: Analyst, 2001,126, 1942-1946
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    A new electrochemical enzyme-linked immunosorbent assay for the screening of macrolide antibiotic residues in bovine meat

    R. Draisci, F. delli Quadri, L. Achene, G. Volpe, L. Palleschi and G. Palleschi, Analyst, 2001, 126, 1942
    DOI: 10.1039/B104939A

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