Abstract：
Combating antimicrobial resistance is a top priority worldwide involving a concerted action by several high-level institutions and organisations in the health sector. To ensure that a meaningful progress is achieved, several campaigns and political initiatives have been launched targeting the health professionals, the industry, the farmers, and the general public. The Regulation (EU) 2019/4 on medicated feed contains provisions for the limitation and control of the contamination of non-target compound feed with 24 antimicrobials. The purpose of this work was to develop a reliable and effective method for the determination of four aminoglycoside antibiotics (apramycin, paromomycin, tobramycin and neomycin) and spectinomycin in feed at cross-contamination level, where an absolute lack of suitable methods was identified. Four candidate methods described in the literature failed to provide adequate recoveries of all analytes. Therefore, an in-depth investigation was carried out to identify the bottleneck variable. The optimised method was then in-house validated and showed performance features appropriate for the intended purpose. The selected compounds could be analysed by LC-MS/MS in five animal feeds with LOQs between 2.6 and 9.2 μg kg-1 for the AGs and between 28 and 86 μg kg-1 for spectinomycin. Using isotopically labelled internal standards, the recovery rates varied from 63 % to 103 % and the intermediate precision (RSDip) varied from 1.1 % to 14 %. This work represents a step forward in the reliable determination of antibiotics in compound feed as the developed method has shown to be precise and sensitive. It is expected that this method gains wide acceptance and can supplement the legislation with effective control tools for antibiotic residues.
摘要：
与抗菌素耐药性作斗争是全球范围内的首要任务，涉及卫生部门的几个高级机构和组织的一致行动。为了确保取得有意义的进展，针对卫生专业人员发起了几项运动和政治倡议，行业,农民,和普通公众。关于含药饲料的法规（EU）2019/4包含限制和控制24种抗菌剂污染非目标复合饲料的规定。这项工作的目的是开发一种可靠有效的方法，用于测定四种氨基糖苷类抗生素（阿普霉素，巴龙霉素,饲料中的妥布霉素和新霉素)和壮观霉素处于交叉污染水平，确定绝对缺乏合适的方法。文献中描述的四种候选方法未能提供所有分析物的足够回收率。因此,进行了深入的调查，以确定瓶颈变量。然后对优化的方法进行内部验证，并显示出适合预期目的的性能特征。可以通过LC-MS/MS分析五种动物饲料中的所选化合物，其中AG的LOQ为2.6至9.2μgkg-1，壮观霉素的LOQ为28至86μgkg-1。使用同位素标记的内标，回收率从63%到103%不等，中间精密度(RSDip)从1.1%到14%不等。这项工作代表了在可靠测定复合饲料中抗生素方面迈出的一步，因为所开发的方法已证明是精确和灵敏的。预计该方法将获得广泛接受，并可以通过有效的抗生素残留控制工具来补充立法。
