Simple and efficient UPLC-ESI-MS/MS method for multi-residue analysis of 14 coccidiostatic agents in poultry liver and muscle tissues

Abstract

Simple, rapid, efficient and cost-effective UPLC-ESI-MS/MS confirmatory method for the simultaneous determination of 14 coccidiostatic agents (salinomycin, monensin, lasalocid, halofuginone, robenidine, nicarbazin as 4,4′-dinitrocarbanilide, narasin, maduramycin, decoquinate, diclazuril, semduramycin, clopidol, amprolium and toltrazuril as parent drug and as its metabolites toltrazuril sulfone and toltrazuril sulfoxide) in chicken and turkey meat and liver was developed, validated and approved in routine laboratory practice. Main steps of sample preparation are as follows: homogenization, ultrasonication with acetonitrile and buffer solution, centrifugation, extraction by organic solvents mixture containing ethyl acetate and dichloromethane, evaporation and defatting of reconstituted residue by hexane. The technique is capable for multi-analyte determination of coccidiostats with LODs of 1–3 μg kg−1 depending on analyte using matrix-fortified calibrations. Procedure's adequacy was evaluated according to Commission Implementing Regulation (EU) 2021/808. Proposed method's efficiency is provided by the rational selection of extractants and ultracentrifugation at last stage of sample preparation. The developed technique is suitable to be used, when applying external standards and internal standards approaches. Time-consuming and costly procedures with ion suppressive desalting agents or SPE purification, which are common for coccidiostats assay, are not involved in the elaborated method, being method's main benefit comparing with known multi-residue LC-MS/MS techniques for coccidiostats analysis. This method is suitable for the control of safety parameters of poultry meat and liver as well as for establishing withdrawal periods of veterinary drugs based on coccidiostatic agents.