Simultaneous determination of quinolones in livestock and marine products by high performance liquid chromatography

Abstract

A high performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of nalidixic acid, flumequine, oxolinic acid, piromidic acid, danofloxacin, enrofloxacin and sarafloxacin in chicken, pork and fish. These seven quinolone antibacterials were extracted with 0.3% metaphosphoric acid: acetonitrile (1:10, v/v), followed by a Bond Elut C18 cartridge clean up procedure. The HPLC separation was achieved on a Cosmosil 5C18-AR-II column (5 μm, 4.6 mm i.d. × 250 mm) with acetonitrile: 0.05M NaH2PO4 (pH 2.5)(35:65, v/v) containing 3.5 mM sodium dodecyl sulfate as a mobile phase, and detection was performed with photodiode array and fluorescence detector (by using wavelength programming). Good linearity was observed from the calibration plot at concentrations from 0.05 to 10.0 mg/mL (0.005-2.0 μg/mL for danofloxacin). Recovery studies of the analytes were performed at 0.01, 0.1, 0.4 and 2.0 ppm (0.001, 0.01, 0.04 and 0.20 ppm for danofloxacin) spiked levels. Average recoveries of low concentration ranged from 74.3 to 85.5% and those of the rest ranged from 80.1 to 99.9% with coefficients of variation less than 5.8%. The detection limits of quinolones were 0.01-0.04 ppm with photodiode array detection and 0.0006-0.05 ppm with fluorescence detection. The coefficients of variation of intra-day and inter-day assays were lower than 3.29% and 5.23%, respectively. These results indicated that the developed method had an acceptable precision. Using this method to detect quinolones in sixty three samples purchased from various markets in Taipei, we found that nine wu ku chicken muscles contained enrofloxacin residues ranged from 0.08 to 4.04 ppm, four wu ku chicken liver muscles contained enrofloxacin residues ranged from 0.01 to 0.27 ppm, and oxolinic acid residues in three sweet fish samples ranged from 0.13 to 0.35 ppm. The results indicated that 25.4% of samples violated the regulation set by the Department of Health. The enrofloxacin residue in wu ku chicken muscle is very stable even after a long period of refrigeration at 4°C.