Carretero Vanesa, Blasco Cristina, Picó Yolanda
Laboratori de Bromatologia i Toxicologia, Facultat de Farmàcia, Universitat de València, Av. Vicent Andrés Estellés s/n, 46100 Burjassot, València, Spain.
J Chromatogr A. 2008 Oct 31;1209(1-2):162-73. doi: 10.1016/j.chroma.2008.09.011. Epub 2008 Sep 7.
A multi-residue method using pressurized liquid extraction (PLE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for determining trace levels of 31 antimicrobials, including beta-lactams, lincosamides, macrolides, quinolones, sulfonamides, tetracyclines, nitroimidazoles and trimethoprim. The extraction method required pre-homogeneization of the meat with EDTA-washed sand and subsequent one-static-cycle extraction for 10 min with 40 ml of water at 1500 psi and 70 degrees C. The effect of operation temperature, pressure, flush volume, and static cycles on PLE performance was studied. Average recoveries ranged from 75 to 99% with relative standard deviations <18%. The method was validated according to the European Union requirements (2002/657/EC). In addition to the quality parameters included in that decision, the limits of detection (LODs) and quantification (LOQs) were determined. The use of LC-MS/MS provided LODs (between 3 and 15 microg kg(-1)) and LOQs (between 10 and 50 microg kg(-1)), by far lower than half of their maximum residue limits (MRLs) (between 50 and 1200 microg kg(-1)). Confirmation of the presence of any of the studied compounds was accomplished in 1h after sample receipt. This methodology has been successfully applied to the analysis of cattle and pig tissue samples from local markets and slaughterhouses of the Valencian Community (Spain). The results showed the presence of some antimicrobials at different concentrations. Quinolones and tetracyclines were the antimicrobials most detected in cattle and pig samples, respectively. Sulfonamides were also frequently detected in both types of samples.
已开发出一种采用加压液体萃取（PLE）和液相色谱 - 串联质谱（LC-MS/MS）的多残留方法，用于测定31种抗菌药物的痕量水平，这些抗菌药物包括β-内酰胺类、林可酰胺类、大环内酯类、喹诺酮类、磺胺类、四环素类、硝基咪唑类和甲氧苄啶。萃取方法要求将肉与经乙二胺四乙酸（EDTA）洗涤的沙子进行预均质化处理，随后在1500磅力/平方英寸和70摄氏度下用40毫升水进行一次静态循环萃取10分钟。研究了操作温度、压力、冲洗体积和静态循环对PLE性能的影响。平均回收率在75%至99%之间，相对标准偏差<18%。该方法根据欧盟要求（2002/657/EC）进行了验证。除了该决议中包含的质量参数外，还测定了检测限（LOD）和定量限（LOQ）。使用LC-MS/MS提供的检测限（在3至15微克/千克之间）和定量限（在10至50微克/千克之间），远低于其最大残留限量（MRL）（在50至1200微克/千克之间）的一半。在收到样品后1小时内完成了对任何一种所研究化合物存在情况的确认。该方法已成功应用于分析来自西班牙巴伦西亚自治区当地市场和屠宰场的牛和猪组织样本。结果表明存在不同浓度的一些抗菌药物。喹诺酮类和四环素类分别是在牛和猪样本中检测到最多的抗菌药物。磺胺类在这两种类型的样本中也经常被检测到。
