Moretti Simone, Cruciani Gabriele, Romanelli Sara, Rossi Rosanna, Saluti Giorgio, Galarini Roberta
Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, Via G. Salvemini, 1, 06126, Perugia, Italy.
Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 10, 06123, Perugia, Italy.
J Mass Spectrom. 2016 Sep;51(9):792-804. doi: 10.1002/jms.3834.
A multiclass method for screening and confirmatory analysis of antimicrobial residues in milk has been developed and validated. Sixty-two antibiotics belonging to ten different drug families (amphenicols, cephalosporins, lincosamides, macrolides, penicillin, pleuromutilins, quinolones, rifamycins, sulfonamides and tetracyclines) have been included. After the addition of an aqueous solution of EDTA, the milk samples were extracted twice with acetonitrile, evaporated and dissolved in ammonium acetate. After centrifugation, 10 µl were analysed using LC-Q-Orbitrap operating in positive electrospray ionization mode. The method was validated in bovine milk in the range 2-150 µg kg(-1) for all antibiotics; for four compounds with maximum residue limits higher than 100 µg kg(-1) , the validation interval has been extended until 333 µg kg(-1) . The estimated performance characteristics were satisfactory complying with the requirements of Commission Decision 2002/657/EC. Good accuracies were obtained also taking advantage from the versatility of the hybrid mass analyser. Identification criteria were achieved verifying the mass accuracy and ion ratio of two ions, including the pseudomolecular one, where possible. Finally, the developed procedure was applied to 13 real cases of suspect milk samples (microbiological assay) confirming the presence of one or more antibiotics, although frequently, the maximum residue limits were not exceeded. The availability of rapid multiclass confirmatory methods can avoid wastes of suspect, but compliant, raw milk samples. Copyright © 2016 John Wiley & Sons, Ltd.
已开发并验证了一种用于牛奶中抗菌药物残留筛查和确证分析的多类别方法。该方法涵盖了属于十个不同药物家族的62种抗生素（氯霉素类、头孢菌素类、林可酰胺类、大环内酯类、青霉素类、截短侧耳素类、喹诺酮类、利福霉素类、磺胺类和四环素类）。加入乙二胺四乙酸（EDTA）水溶液后，牛奶样品用乙腈萃取两次，蒸发后溶解于醋酸铵中。离心后，取10μl样品采用正电喷雾电离模式运行的液相色谱 - 四极杆 - 轨道阱质谱仪（LC - Q - Orbitrap）进行分析。该方法在牛奶中对所有抗生素的验证范围为2 - 150μg kg⁻¹；对于四种最大残留限量高于100μg kg⁻¹的化合物，验证区间扩展至333μg kg⁻¹。估计的性能特征令人满意，符合欧盟委员会第2002/657/EC号决定的要求。利用混合质量分析仪的多功能性也获得了良好的准确度。通过验证两个离子（尽可能包括准分子离子）的质量准确度和离子比率来达到鉴定标准。最后，将所开发的程序应用于13个疑似牛奶样品的实际案例（微生物检测），证实存在一种或多种抗生素，不过通常最大残留限量未被超过。快速多类别确证方法的可用性可以避免浪费可疑但合格的原料奶样品。版权所有© 2016约翰威立父子有限公司。
