Development and application of a dispersive liquid–liquid microextraction method for the determination of tetracyclines in beef by liquid chromatography mass spectrometry

Abstract

A rapid, cost effective and environmentally friendly extraction method, based on dispersive liquid–liquid microextraction (DLLME) was developed for the determination of six tetracyclines in meat destined for human consumption. Meat extracts were analysed for tetracyclines using liquid chromatography tandem mass spectrometry (LC–MS/MS), a sensitive and selective analytical technique. Various factors influencing the pre-concentration of tetracyclines such as sample pH, type and volume of both disperser solvent and extraction solvent were optimized. Validation parameters such as calibration function, limit of detection (LOD), limit of quantification (LOQ), detection capability (CCα), decision limit (CCβ), accuracy and precision were established according to Commission Decision 2002/657/EC. Linearity in the range of 25–200µgkg−1was obtained with regression coefficients ranging from 0.9991 to 0.9998. Recoveries of spiked blank muscle samples at three levels (50, 100 and 150µgkg−1) ranged from 80% to 105% and reproducibility was between 2% and 7%. LODs and LOQs ranged from 2.2 to 3.6µgkg−1 and from 7.4 to 11.5µgkg−1 respectively while CCα ranged from 105 to 111µgkg−1 and CCβ ranged from 107 to 122µgkg−1. The proposed method compared well with the existing accepted dispersive solid phase extraction method and was successfully applied to the pre-concentration and determination of tetracyclines in meat samples. Eleven of the thirty bovine muscle samples obtained from local abattoirs and butcheries were found to contain residues of two tetracycline antibiotics (chlortetracycline and oxytetracycline), with oxytetracycline being the most detected. Concentration levels of the tetracycline residues detected in bovine muscle samples were lower (12.4 and 68.9µgkg−1) than the stipulated European Union maximum residue level (MRL) of 100µgkg−1, hence the meat was fit for human consumption. From this work it can be concluded that the DLLME is indeed a greener sample preparation method and could be used as an alternative to dispersive solid phase microextraction (dSPE).