Simultaneous Determination of Tetracyclines and Fluoroquinolones in Poultry Eggs by UPLC Integrated with Dual-Channel-Fluorescence Detection Method

Abstract

An innovative, rapid and stable method for simultaneous determination of three tetracycline (oxytetracycline, tetracycline and doxycycline) and two fluoroquinolone (ciprofloxacin and enrofloxacin) residues in poultry eggs by ultra-high performance liquid chromatography-fluorescence detection (UPLC-FLD) was established and optimized. The samples were homogenized and extracted with acetonitrile/ultrapure water (90:10, v/v) and then purified by solid-phase extraction (SPE). LC separation was achieved on an ACQUITY UPLC BEH C18 column (1.7 µm, 2.1 mm × 100 mm), and the mobile phase was composed of acetonitrile and a 0.1 mol/L malonic acid solution containing 50 mmol/L magnesium chloride (the pH was adjusted to 5.5 with ammonia). When the five target drugs were spiked at the limit of quantification, 0.5 times the maximum residue limit (MRL), 1.0 MRL and 2.0 MRL, the recoveries were above 83.5% and the precision ranged from 1.99% to 6.24%. These figures of merit complied with the parameter validation regulations of the EU and U.S. FDA. The limits of detection and quantifications of the targets were 0.1-13.4 µg/kg and 0.3-40.1 µg/kg, respectively. The proposed method was easily extended to quantitative analyses of target drug residues in 85 egg samples, thus demonstrating its reliability and applicability.

Keywords: UPLC-FLD; dual-channel; fluoroquinolones; poultry eggs; tetracyclines.

Figure 1

Chromatograms of the mobile phase (a), 200 µg/kg OTC, TC and DOX standards (b), blank blended whole chicken egg (c) and blank blended whole chicken egg spiked with 200 µg/kg OTC, TC and DOX standards (d) from channel A.

Figure 2

Chromatograms of the mobile phase (a), 50 µg/kg CIP and 25 µg/kg ENR standards (b), blank blended whole chicken egg (c) and blank blended whole chicken egg spiked with 50 µg/kg CIP and 25 µg/kg ENR standards (d) from channel B.

Figure 3

Chromatograms of the mobile phase (a), 200 µg/kg OTC, TC and DOX standards (b), blank chicken albumen (c) and blank chicken albumen spiked with 200 µg/kg OTC, TC and DOX standards (d) from channel A.

Figure 4

Chromatograms of the mobile phase (a), 50 µg/kg CIP and 25 µg/kg ENR standards (b), blank chicken albumen (c) and blank chicken albumen spiked with 50 µg/kg CIP and 25 µg/kg ENR standards (d) from channel B.

Figure 5

Chromatograms of the mobile phase (a), 200 µg/kg OTC, TC and DOX standards (b), blank chicken yolk (c) and blank chicken yolk spiked with 200 µg/kg OTC, TC and DOX standards (d) from channel A.

Figure 6

Chromatograms of the mobile phase (a), 50 µg/kg CIP and 25 µg/kg ENR standards (b), blank chicken yolk (c) and blank chicken yolk spiked with 50 µg/kg CIP and 25 µg/kg ENR standards (d) from channel B.