Development and validation according to European Union Decision 2002/657/EC of an HPLC-DAD method for milk multi-residue analysis of penicillins and amphenicols based on dispersive extraction by QuEChERS in MSPD format

摘要

A precise and reliable method for milk residue analysis regarding five penicillins and three amphenicols by HPLC-diode array detection has been developed herein. The chromatographic separation was performed using a mobile phase of CH _3COONH _4 (0.05 M) and ACN delivered by gradient program on a Kinetex ~(a￠)-C _(18) core-shell, 2.6 μm column, starting at a volume ratio of 95:5 and ending at 60:40 after 17 min, remaining stable for 3 more min. A modified matrix solid phase dispersion procedure was applied for the extraction and clean-up procedure of antibiotics using a mixture of Strata by Phenomenex and QuEChERS as a sorbent. The method was validated at the respective 0.5 × MRL, MRL and 1.5 ? - MRL level for each compound. Results were quantitated against the internal standard paracetamol (2 ng/μL) according to the matrix-matched approach. The method was validated in line with the EC guidelines as cited in the Decision 2002/657/EC. The within-laboratory reproducibility, expressed as a RSD, never exceeded 16%. All decision limit (CCα) values lay in the range between 35.2 and 56.3 μg/kg and the corresponding results for detection capability (CCβ) were 39.9 and 61.9 μg/kg. Ruggedness was estimated according to the Youden approach.