Agilent Technologies, 2850 Centerville Road, Wilmington, DE 19808, USA.
Agilent Technologies, 2850 Centerville Road, Wilmington, DE 19808, USA.
J Chromatogr A. 2018 May 11;1549:14-24. doi: 10.1016/j.chroma.2018.03.033. Epub 2018 Mar 17.
This study presents the development and validation of a quantitation method for the analysis of multi-class, multi-residue veterinary drugs using lipid removal cleanup cartridges, enhanced matrix removal lipid (EMR-Lipid), for different meat matrices by liquid chromatography tandem mass spectrometry detection. Meat samples were extracted using a two-step solid-liquid extraction followed by pass-through sample cleanup. The method was optimized based on the buffer and solvent composition, solvent additive additions, and EMR-Lipid cartridge cleanup. The developed method was then validated in five meat matrices, porcine muscle, bovine muscle, bovine liver, bovine kidney and chicken liver to evaluate the method performance characteristics, such as absolute recoveries and precision at three spiking levels, calibration curve linearity, limit of quantitation (LOQ) and matrix effect. The results showed that >90% of veterinary drug analytes achieved satisfactory recovery results of 60-120%. Over 97% analytes achieved excellent reproducibility results (relative standard deviation (RSD) < 20%), and the LOQs were 1-5 μg/kg in the evaluated meat matrices. The matrix co-extractive removal efficiency by weight provided by EMR-lipid cartridge cleanup was 42-58% in samples. The post column infusion study showed that the matrix ion suppression was reduced for samples with the EMR-Lipid cartridge cleanup. The reduced matrix ion suppression effect was also confirmed with <15% frequency of compounds with significant quantitative ion suppression (>30%) for all tested veterinary drugs in all of meat matrices. The results showed that the two-step solid-liquid extraction provides efficient extraction for the entire spectrum of veterinary drugs, including the difficult classes such as tetracyclines, beta-lactams etc. EMR-Lipid cartridges after extraction provided efficient sample cleanup with easy streamlined protocol and minimal impacts on analytes recovery, improving method reliability and consistency.
本研究开发并验证了一种使用脂质去除净化管（EMR-Lipid），通过液相色谱串联质谱检测，对不同肉类基质中的多类多残留兽药进行定量分析的方法。采用两步固相-液相提取，然后进行穿透式样品净化，对肉样进行提取。该方法基于缓冲液和溶剂组成、溶剂添加剂添加以及 EMR-Lipid 净化管进行优化。然后，在五种肉类基质（猪肌肉、牛肌肉、牛肝、牛肾和鸡肝）中对开发的方法进行验证，以评估方法性能特征，如三个加标水平的绝对回收率和精密度、校准曲线线性、定量限（LOQ）和基质效应。结果表明，>90%的兽药分析物的回收率达到 60-120%的满意水平。超过 97%的分析物表现出良好的重现性（相对标准偏差（RSD）<20%），在评估的肉类基质中的 LOQ 为 1-5μg/kg。EMR-Lipid 净化管净化时，基质共提取去除效率为 42-58%。柱后注入研究表明，EMR-Lipid 净化管可减少样品中的基质离子抑制。在所有肉类基质中，所有测试的兽药中，具有显著定量离子抑制（>30%）的化合物频率<15%，也证实了这种基质离子抑制效应的降低。结果表明，两步固相-液相提取为包括四环素、β-内酰胺等困难类别在内的整个兽药谱提供了有效的提取。提取后的 EMR-Lipid 净化管可提供高效的样品净化，具有简单的流线型方案，对分析物回收率的影响最小，从而提高方法的可靠性和一致性。
