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HPLC confirmatory method development for the determination of seven quinolones in salmon tissue (Salmo salar L.) validated according to the European Union Decision 2002/657/EC

Evaggelopoulou, Evaggelia N., Samanidou, Victoria F.
Food chemistry 2013 v.136 no.2 pp. 479-484
European Union, Salmo salar, antibiotics, buffers, ciprofloxacin, citrates, danofloxacin, detectors, enrofloxacin, flumequine, high performance liquid chromatography, methanol, nalidixic acid, oxolinic acid, salmon, sarafloxacin, solid phase extraction
A confirmatory high pressure liquid chromatographic method for the determination of seven quinolone antibiotics in tissue of Atlantic salmon (Salmo salar L.) was developed. Ciprofloxacin (CIP), danofloxacin (DAN), enrofloxacin (ENR), sarafloxacin (SAR), oxolinic acid (OXO), nalidixic acid (NAL) and flumequine (FLU) were separated on a Perfectsil ODS-2 120 (250mm×4mm, 5μm) column by gradient elution with a mobile phase consisting of 0.1% trifluoroacetic acid (pH=1), acetonitrile and methanol at 25°C within 22min. Analytes were monitored at 255nm (for the determination of OXO, NAL and FLU) and 275nm (for CIP, DAN, ENR and SAR) by means of photodiode array detector. Examined quinolones were isolated from salmon tissue by extraction with citrate buffer solution (pH=4.7) and purified by solid phase extraction using Oasis HLB (200mg/6mL) cartridges. The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability and sensitivity according to the European Union Decision 2002/657/EC. The accuracy of the method was additionally proved by its application to certified reference material of salmon tissue (BCR® 725).