Authors
Siji Joseph, Aimei Zou, Chee Sian Gan, Limian Zhao, and Patrick Batoon Agilent Technologies, Inc.
A comprehensive LC/MS/MS workflow was developed for targeted screening or quantitation of 210 veterinary drug residues in animal muscle prepared for human consumption, with the intention to accelerate and simplify routine laboratory testing. The workflow ranged from sample preparation through chromatographic separation, MS detection, data processing and analysis, and report generation. The workflow performance was evaluated using three muscle matrices—chicken, pork, and beef— and was assessed on two different Agilent triple quadrupole LC/MS models (an Agilent 6470 and a 6495C triple quadrupole LC/MS). A simple sample preparation protocol using Agilent Captiva EMR—Lipid cartridges provided efficient extraction and matrix cleanup. A single chromatographic method using Agilent InfinityLab Poroshell 120 EC-C18 columns with a 13-minute method delivered acceptable separation and retention time distribution across the elution window for reliable triple quadrupole detection and data analysis.
Workflow performance was evaluated based on evaluation of limit of detection (LOD), limit of quantitation (LOQ), calibration curve linearity, accuracy, precision, and recovery, using matrix-matched spike samples for a range from 0.1 to 100 μg/L. Calibration curves were plotted from LOQ to 100 μg/L, where all analytes demonstrated linearity R2 >0.99. Instrument method accuracy values were within 73 to 113%. Target analytes response and retention time %RSD values were ≤19% and ≤0.28% respectively. Analyte recovery and reproducibility at three levels of fortified quality control (QC) samples—1, 10, and 25 μg/kg in meat—were used to validate the method applicability for confident routine screening of veterinary drugs. The recovery repeatability (intrabatch technical replicates) and recovery reproducibility (interbatch technical replicates) were calculated using QC samples, and the results were within acceptable limits of 20 and 32%, respectively.1 The workflow method performance results across the chicken, beef, and pork muscle matrices showed excellent overlap, and confirm the method applicability for routine multiresidue screening in various animal origin matrices.
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