Development of a Direct Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Fluoroquinolone Residues in S
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Development of a Direct Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Fluoroquinolone Residues in Shrimp Sheryl A. Tittlemier & Jean-Marc Gélinas & Guy Dufresne & Mehul Haria & Jessica Querry & Chantal Cleroux & Cathie Ménard & Philippe Delahaut & Gurmit Singh & Nathalie Fischer-Durand & Samuel Benrejeb Godefroy
Received: 25 September 2007 / Accepted: 12 November 2007 / Published online: 29 December 2007 # Springer Science + Business Media, LLC 2007
Abstract Rabbit polyclonal antibodies were generated against norfloxacin, purified, and used as the basis of a direct competitive enzyme-linked immunosorbent assay for the screening of fluoroquinolones in shrimp. The developed method used a simple ethanol/acetic acid solvent extraction, which resulted in a 1.0-ng-norfloxacin/g limit of detection (based on the analysis of known negative and fortified shrimp samples). Norfloxacin extraction efficiencies were evaluated at two fortification concentrations and were greater than 70%, with an intra-assay variation less than 30%. The assay displayed greater than 10% cross reactivity against enro-, cipro-, sara-, and difloxacin. Incurred and known negative shrimp samples were analyzed and S. A. Tittlemier : J.-M. Gélinas : M. Haria : J. Querry : C. Cleroux : C. Ménard : G. Singh : S. B. Godefroy Bureau of Chemical Safety, Food Directorate, Health Canada, Ottawa, ON K1A 0L2, Canada G. Dufresne Quebec Region Laboratory, Health Canada, Longueuil, QC J4K 1C7, Canada P. Delahaut Laboratoire d’Hormonologie, Centre d’Economie Rurale, 8 Rue Point du Jour, 6900 Marloie, Belgium N. Fischer-Durand Laboratoire de Chimie et Biochimie des Complexes Moléculaires, Ecole Nationale Supérieure de Chimie de Paris, 75005 Paris, France S. A. Tittlemier (*) Food Research Division, Health Canada, Ottawa, ON K1A 0L2, Canada e-mail: [email protected]
compared to the results obtained from an independent liquid chromatography tandem mass spectrometric method. All three instances in which fluoroquinolones were present at concentrations near or above the assay limit of detection (1.0 to 17 ng/g) were identified as positive by the newly developed assay, demonstrating the usefulness of this assay as a screening tool. Keywords ELISA . Veterinary Drug . Antibiotic . Aquaculture . Food . Antibodies . Fluoroquinolones
Introduction Fluoroquinolones are a class of synthetic antibiotics with both human and veterinary medical uses. In Canada, only two fluoroquinolones are permitted for use in food animals. Danofloxacin and enrofloxacin are approved for use in beef cattle to treat bovine respiratory disease and have administrative maximum residue limits set at 0.4 μg/g in kidney and 0.02 μg/g in muscle, respectively. Fluoroquinolones are not approved for use in fish and shellfish in Canada, and the presence of these compounds in products imported from other countries into Canada is not allowed. Because there are indications that fluoroquinolones have been used in shrimp farming in the past (Saitanu et al. 1994, Holmstrom et al. 2003),
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