Development and Validation of an HPLC Method for the Determination of Seven Tetracycline Antibiotics Residues in Chicken Muscle and Egg Yolk According to 2002/657/EC

Abstract

Herein, an HPLC method with diode-array detection, at 355 nm, is described for the determination of seven tetracyclines in chicken muscle and egg yolk: minocycline, tetracycline, oxytetracycline, methacycline, demeclocycline, chlortetracycline, and doxycycline. The examined TCs were extracted from chicken muscle with 15% TFA and 0.4 M oxalate buffer (pH 4) and from egg yolk with 40% TFA and 0.4 M oxalate buffer (pH 4). LiChrolut and Nexus SPE cartridges were used for further purification of chicken tissue and egg yolk extracts, respectively, with 0.01 M C 2 H 2 O 4 /CH 3 CN/CH 3 OH (40:30:30 v/v/v) as elution solvent.The separation was achieved on a Kromasil ODS-3, 5 m, 250 × 4 mm, analytical column. The mobile phase, a mixture of 0.01 M oxalic acid and CH 3 CN, was delivered using a gradient program. The procedure was validated according to the Decision 2002/657/EC, determining selectivity, stability, decision limit, detection capability, accuracy, and precision. Overall recoveries ranged from 92-110.1% and 89-106% for chicken tissues and eggs, respectively. All RSD values were lower than 11%. The decision limits CC a in chicken tissues ranged from 102.98 to 109.11 g/kg, while detection capability CC b from 113.45 to 118.18 g/kg. Respective values in eggs were 206.53-214.60 g/kg for CC a and 216.21-228.97 g/kg for CC b .

Ultrasound-assisted dispersive liquid–liquid microextraction of tetracycline drugs from egg supplements before flow injection analysis coupled to a liquid waveguide capillary cell

“…A reference HPLC-DAD technique was used for comparison with the results obtained by the proposed methodology, as described by Konstantina et al [11]. After extraction of the tetracyclines from the egg samples, a solid-phase extraction purification procedure was performed.…”

“…The analytical techniques reported in the literature for the monitoring and determination of tetracycline antibiotics and other veterinary drugs in eggs are mostly chromatographic, including high performance liquid chromatography (HPLC) with ultraviolet (UV) [7,[11][12][13], fluorescence [14], or mass spectrometry (MS/MS) [1][2][3][4][15][16][17][18][19][20] detection, and ultrahigh-pressure liquid chromatography-tandem mass spectrometry (UHPL-MS/MS) [5,6].…”

“…The most important are solvent extraction [5,16], liquid-liquid extraction (LLE) [3], solidphase extraction (SPE) [1,2,4,5,11,14,15,17,19], ultrasonic-assisted extraction (UAE) [2], and pressurized liquid extraction (PLE) [6,12]. Furthermore, SPE modifications using molecularly imprinted solid-phase extraction (MISPE) [13,20] and magnetic molecularly imprinted polymer solidphase extraction (MIP-SPE) [18] have also been used as extraction methods.…”

Effect of probiotics on doxycycline disposition in gastro-intestinal tract of poultry

“…1308100010) preconditioned with 2 mL methanol and 2 mL water. Elution was performed with a mixture of methanol/acetonitrile/0.01M oxalic acid according to Nikolaidou et al (2008). The eluents were evaporated to dryness at 40 °С and the dry residues were reconstituted in 100 μL of methanol; then 20 μL were injected into the HPLC system (Nikolaidou et al, 2008).…”

“…Elution was performed with a mixture of methanol/acetonitrile/0.01M oxalic acid according to Nikolaidou et al (2008). The eluents were evaporated to dryness at 40 °С and the dry residues were reconstituted in 100 μL of methanol; then 20 μL were injected into the HPLC system (Nikolaidou et al, 2008). Standard curves were prepared with tissue samples from untreated animals and solutions of doxycycline with concentration 20, 10, 5, 2.5 and 1 μg/mL.…”