Ultrasound-assisted dispersive extraction for the high pressure liquid chromatographic determination of tetracyclines residues in milk with diode array detection

Abstract

Ultrasound assisted matrix solid phase dispersive extraction was applied for the selective isolation and clean-up of tetracyclines (oxytetracycline, tetracycline, epi-chlorotetracycline, chlorotetracycline and doxycycline) from milk. Target analytes were determined by an accurate and sensitive chromatographic analytical method, which was validated to meet the European Legislation criteria. The separation was performed on a LiChroCART-LiChrospher® 100 RP-18 (5μm, 250×4mm) analytical column, operated at ambient temperature, followed by diode array detection.Validation included investigation of linearity, selectivity, stability, limits of detection and quantitation, decision limit, detection capability, trueness, precision and ruggedness according to the Youden’s approach. Limits of quantitation of examined tetracyclines were from 14.5 to 56.6μg/kg significantly lower than respective Maximum Residue Limits, whereas recoveries ranged from 82.0% to 108%. The applicability of the method was evaluated using milk samples purchased from local market. Accuracy of the method was additionally proved by analysis of bovine milk certified reference material (BCR®-492).