RIKILT, Wageningen University and Research Centre, Akkermaalsbos 2, 6708WB, P.O. Box 230, 6700AE, Wageningen, The Netherlands,
Anal Bioanal Chem. 2013 Sep;405(24):7859-74. doi: 10.1007/s00216-013-6804-6. Epub 2013 Feb 22.
A comprehensive method for the quantitative residue analysis of trace levels of 22 ß-lactam antibiotics, including penicillins, cephalosporins, and carbapenems, in poultry muscle by liquid chromatography in combination with tandem mass spectrometric detection is reported. The samples analyzed for ß-lactam residues are hydrolyzed using piperidine in order to improve compound stability and to include the total residue content of the cephalosporin ceftifour. The reaction procedure was optimized using a full experimental design. Following detailed isotope labeling, tandem mass spectrometry studies and exact mass measurements using high-resolution mass spectrometry reaction schemes could be proposed for all ß-lactams studied. The main reaction occurring is the hydrolysis of the ß-lactam ring under formation of the piperidine substituted amide. For some ß-lactams, multiple isobaric hydrolysis reaction products are obtained, in accordance with expectations, but this did not hamper quantitative analysis. The final method was fully validated as a quantitative confirmatory residue analysis method according to Commission Decision 2002/657/EC and showed satisfactory quantitative performance for all compounds with trueness between 80 and 110% and within-laboratory reproducibility below 22% at target level, except for biapenem. For biapenem, the method proved to be suitable for qualitative analysis only.
本文报道了一种用于检测家禽肌肉中痕量 22 种β-内酰胺类抗生素(包括青霉素类、头孢菌素类和碳青霉烯类)残留的综合定量分析方法。该方法采用哌啶对样品进行水解,以提高化合物稳定性并包括头孢曲松的总残留含量。通过全因子实验设计对反应条件进行了优化。在详细的同位素标记、串联质谱研究和使用高分辨率质谱反应方案进行精确质量测量后,可以为所有研究的β-内酰胺类抗生素提出反应方案。主要反应是β-内酰胺环的水解,生成哌啶取代的酰胺。对于一些β-内酰胺类抗生素,根据预期得到了多种等质量的水解反应产物,但这并不妨碍定量分析。最终方法完全按照委员会决议 2002/657/EC 进行了定量确证残留分析方法验证,除比阿培南外,所有化合物的准确度在 80%至 110%之间,目标水平下的实验室内重复性低于 22%,均表现出满意的定量性能。对于比阿培南,该方法仅被证明适用于定性分析。