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An enzyme-linked immunosorbent assay for the determination of chloramphenicol using a monoclonal antibody Application to residues in swine muscle tissue

Die Anwendung monoklonaler Antikörper bei der Bestimmung von Choramphenicol-Rückständen mit der ELISA-Methode Anwendung bei Rückständen im Schweinefleisch

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Zusammenfassung

Es wird eine kompetitive Methode „ELISA” (enzyme-linked immunosorbent assay) mit monoklonalen Antikörpern zur Bestimmung von Chloramphenicol-Rückständen (CAP) in Schweinefleisch beschrieben. Für Standardlösungen wurde als Nachweisgrenze 25 ng ml−1 festgestellt (=2.0 ng CAP). Die sehr große Spezifität dieser monoklonalen Antikörper für CAP drückt sich durch die unwichtigen Kreuzreaktionen mit anderen antimikrobiellen Substanzen und mit strukturell verwandten Verbindungen aus. Zur Probenaufarbeitung wurde das gleiche Schnellverfahren benutzt wie bei der Hochleistungs-Flüssigchromatographie. Rückstände > 5 μg kg−1 können im Schweinefleisch leicht bestimmt werden. Bei Zusatzmengen von 10, 15, 25 und 50 μg CAP pro kg sind die Wiederfindungsraten gut. Die Ergebnisse mit der ELISA-Methode wurden mit HPLC bestätigt.

Summary

A competitive enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody is described for the determination of chloramphenicol (CAP) residues in swine muscle tissue. The limit of detection of standard solutions is established to be 25 ng ml−1 = 2.0 ng CAP per well). The very high specificity of the monoclonal antibody for CAP is expressed by the insignificant cross-reactivity with other antimicrobial agents and with structurally related compounds. By means of the rapid sample preparation method described earlier for the CAP determination using high-performance liquid chromatography (HPLC), residue levels of CAP in swine muscle tissue above 5 μg kg−1can be easily quantitated. The muscle samples show good recovery percentages at 10–50 μg kg−1 spiking levels. The results obtained by the ELISA method were confirmed by HPLC.

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Authors and Affiliations

  1. Department of the Science of Food of Animal Origin, Section Chemistry, Faculty of Veterinary Medicine, The University of Utrecht, P. O. Box 80175, NL-3508 TD, Utrecht, The Netherlands

    Cornelis van de Water & Nel Haagsma

  2. Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, The University of Utrecht, The Netherlands

    Peter J. S. van Kooten & Willem van Eden

Authors
  1. Cornelis van de Water
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  2. Nel Haagsma
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  3. Peter J. S. van Kooten
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  4. Willem van Eden
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van de Water, C., Haagsma, N., van Kooten, P.J.S. et al. An enzyme-linked immunosorbent assay for the determination of chloramphenicol using a monoclonal antibody Application to residues in swine muscle tissue. Z Lebensm Unters Forch 185, 202–207 (1987). https://doi.org/10.1007/BF01042047

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  • DOI: https://doi.org/10.1007/BF01042047

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