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Rapid Confirmatory Method for the Determination of Danofloxacin and N-Desmethyl Danofloxacin in European Seabass by UPLC-PDA

[ Vol. 14 , Issue. 1 ]


Sofia C. Vardali, Victoria F. Samanidou* and Yannis P. Kotzamanis   Pages 68 - 74 ( 7 )


Background: Danofloxacin is a synthetic antibacterial agent with broad spectrum antibacterial and antimycoplasmal activity which is widely used in aquaculture.

Methods: A UPLC (Ultra Performance Liquid Chromatography) method was developed and validated for the determination of danofloxacin (Dano) and its major metabolite N-desmethyl danofloxacin (Ndesmethyl Dano) in muscle plus skin tissue of European seabass (Dicentrarchus labrax) for the first time. For the separation an analytical column UPLC BEH C18 2.1 × 5 mm (1.7 µm) was used with an isocratic mobile phase consisting of MeOH-water acidified with formic acid (0.01%) (18:82; v/v) and flow rate of 0.3 mL/min within 5.5 min. The detection was performed at 275 nm using a photodiode array detector for both compounds. Examined fluoroquinolones were isolated from seabass muscle and skin tissue by extraction with acidic ACN and further purification with the QuEChERS methodology.

Results: Recovery rates from muscle and skin tissue ranged between 90.2 and 101.2% for both compounds. The detection limit of the method was estimated at 13.73 µg/kg for Dano, 18.32 µg/kg for Ndesmethyl Dano, while the limits of quantification were 41.62 and 55.52 µg/kg, respectively. 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.

Conclusion: The new developed and validated method can be readily applied to aqua cultured fish after dietary administration of danofloxacin.


Danofloxacin, metabolite, fish, seabass, residue analysis, QuEChERS, UPLC-PDA.


Institute of Biological Marine Resources, Hellenic Center of Marine Research, Agios Kosmas, Hellenikon, 16777 Athens, Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki. GR-54 124 Thessaloniki, Institute of Biological Marine Resources, Hellenic Center of Marine Research, Agios Kosmas, Hellenikon, 16777 Athens

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