, Volume 60, Issue 11, pp 657-661

First online:

Determination of Oxolinic Acid Residues in Gilthead Seabream (Sparus aurata) Muscle Tissue and Plasma by High-Performance Liquid Chromatography

  • A. E. TyrpenouAffiliated withLaboratory of High Performance Liquid Chromatography, Institute of Veterinary Research of Athens, National Agricultural Research Foundation Email author 
  • , G. RigosAffiliated withLaboratory of Fish Nutrition and Pathology, Hellenic Centre of Marine Research Agios Kosmas

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A high-performance liquid chromatographic method for the determination of oxolinic acid (OA) residues in muscle tissue and plasma of the cultured fish gilthead seabream (Sparus aurata L.), is described. OA was extracted with ethyl acetate and after centrifugation the combined extracts were evaporated. To the remaining residue 1 mL of the mobile phase was added and the extract was partitioned with n-pentane which then was rejected by aspiration. OA was chromatographed on a Zorbax®SB-C18 column at 50oC and detected by fluorescence detection at λex 327 nm and λem 369 nm. The mobile phase was a mixture of 0.1% trifluoroacetic acid (v/v) pH 2.0 and acetonitrile-methanol 3:2 (v/v) in a combination of 50:50 (v/v) and a flow rate of 1.0 mL min−1, delivered isocratically. Method mean recovery (R%) achieved was 73.7 ± 4.4% (mean ± SD) for blank fortified samples (n=4) range at 50, 100 and 200 μg kg−1 with a RSD=3.3%. The limit of detection (LOD) was 2.0 μg kg−1 oxolinic acid in muscle tissue and plasma and the limit of quantification (LOQ) was 5.0 μg kg−1. The method is fast and suitable to be used with safety and accuracy for the control of OA residues in cultured seabreams and a trained analyst could carry out ready for chromatography more than 50 samples per working day.


Column liquid chromatography Oxolinic acid residues Seabream Farmed fish