a State General Laboratory of Cyprus , Veterinary Drug Residues Laboratory , Nicosia , Cyprus.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2013;30(10):1728-32. doi: 10.1080/19440049.2013.814075. Epub 2013 Jul 22.
A limited number of substances are authorised for the treatment of bees. Maximum residue limits (MRLs) are set for tetracyclines in several matrices, but not for honey. Nevertheless, tetracycline antibiotics may be used in order to prevent bacterial diseases and the loss of honey bee populations. In this study, a sensitive multi-residue LC-MS/MS method was developed and optimised for the quantitative and qualitative determination of tetracycline residues in honey. Homogenisation of samples under acidic conditions was performed and solid-phase extraction was carried out. The eluate was evaporated under nitrogen and dissolved in an aqueous methanol solution prior to filtration. A mobile phase composed of acetic acid-water and acetic acid-acetonitrile was used. Separation of tetracycline, oxytetracycline, chlortetracycline and doxytetracycline was achieved by using gradient elution on a C18 chromatography column. The analytical method was validated according to Commission Decision 2002/657/EC by the analysis of spiked samples around the recommended concentration of 20 μg kg(-1) by EURL Guidance Paper, December 2007. A matrix effect was observed, so quantification was based on an external matrix calibration curve. Calculated decision limits (CCα) were lower than 10 μg kg(-1) for all tetracyclines. Good linearity, repeatability and within-laboratory reproducibility were achieved.
仅有少数物质被批准用于治疗蜜蜂。几种基质中的四环素都设定了最大残留限量（MRL），但蜂蜜中没有。尽管如此，为了预防蜜蜂细菌性疾病和蜂群的减少，仍可能使用四环素类抗生素。在这项研究中，开发并优化了一种用于定量和定性测定蜂蜜中四环素残留的灵敏多残留 LC-MS/MS 方法。在酸性条件下对样品进行均质化，并进行固相萃取。在氮气流下蒸发洗脱液，然后溶解在甲醇水溶液中，再进行过滤。使用由乙酸-水和乙酸-乙腈组成的流动相。在 C18 色谱柱上采用梯度洗脱，实现了四环素、土霉素、金霉素和强力霉素的分离。根据委员会 2002/657/EC 号决定，通过分析 2007 年 12 月 EURL 指导文件中推荐浓度 20μgkg-1 左右的加标样品，对分析方法进行了验证。观察到基质效应，因此定量基于外部基质校准曲线。所有四环素的计算决策限（CCα）均低于 10μgkg-1。达到了良好的线性、重复性和实验室内部再现性。
