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Abstract
A high performance liquid chromatographic method for the determination of chloramphenicol residues in muscle tissue of the cultured fish gilthead seabream (Sparus aurata L.) was developed. Chloramphenicol is extracted with ethyl acetate and after centrifugation and solvent evaporation the oily extract is partitioned between 3% sodium chloride solution and n-pentane, and chloramphenicol is extracted back into ethyl acetate. After evaporation to near dryness, the residue is dissolved in n-hexane and is cleaned up on a Silica gel SPE mini column. Chloramphenicol was analyzed on a ZORBAX SB - C18 column at a temperature of 50°C, with the mobile phase being methanol:water 30 + 70 v/v delivered isocratically. Detection was performed using a Photo Diode Array detector monitored at λmax ∼ 278 nm. The mean recovery (R%) achieved was 88.62 ± 9.65% for a range of 10, 25, 50, 100, 200 μg/kg blank fortified samples (n = 4). The limit of detection (LOD) was 1.87 ng corresponding to 5 μg/kg chloramphenicol in muscle and the limit of quantification (LOQ) was 10 μg/kg.