Development and validation of a streamlined method designed to detect residues of 62 veterinary drugs in bovine kidney using ultra‐high performance liquid chromatography – tandem mass spectrometry

Abstract

In the USA, the US Department of Agriculture's Food Safety and Inspection Service (FSIS) conducts the National Residue Program designed to monitor veterinary drug and other chemical residues in beef and other slaughtered food animals. Currently, FSIS uses a 7-plate bioassay in the laboratory to screen for antimicrobial drugs in bovine kidneys from those animals tested positive by inspectors in the slaughter establishments. The microbial inhibition bioassay has several limitations in terms of monitoring scope, sensitivity, selectivity, and analysis time. Ultra-high performance liquid chromatography -tandem mass spectrometry (UHPLC-MS/MS) has many advantages over the bioassay for this application, and this study was designed to develop, evaluate, and validate a fast UHPLC-MS/MS method for antibiotics and other high-priority veterinary drugs in bovine kidney. Five existing multi-class, multi-residue methods from the literature were tested and compared, and each performed similarly. Experiments with incurred samples demonstrated that a 5-min shake of 2 g homogenized kidney with 10 ml of 4/1 (v/v) acetonitrile/water followed by simultaneous clean-up of the initial extract with 0.5 g C18 and 10 ml hexane gave a fast, simple, and effective sample preparation method for the <10 min UHPLC-MS/MS analysis. An extensive 5-day validation process demonstrated that the final method could be used to acceptably screen for 54 of the 62 drugs tested, and 50 of those met qualitative MS identification criteria. Quantification was not needed in the application, but the method gave ≥70% recoveries and ≤25% reproducibilities for 30 of the drugs. Published 2012. This article is a U.S. Government work and is in the public domain of the USA.

Identification in residue analysis based on liquid chromatography with tandem mass spectrometry: Experimental evidence to update performance criteria

“…Only when one or both product ions are in the low S/N range (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15), higher deviations are more frequently observed. Based on the 95th percentile of the RSDs obtained for the reference ion ratios, deviations up to AE45% (relative) can be expected.…”

“…Operation in multiple-reaction monitoring (MRM) mode permits the measurement of multiple analytes (typically more than 150) in a single analytical run and thus it is not surprising that LC-MS/MS has become the method of choice for routine pesticide multi-residue analysis [1,2]. This trend is also observed in the field of veterinary drugs [3][4][5], mycotoxins [6][7][8], and in food safety and quality analysis in general [9,10]. Similar developments have taken place in other analytical domains including environmental analysis [11], forensic and clinical toxicology [12,13], and in human sports doping [14,15].…”

Evaluation of a multi‐class, multi‐residue liquid chromatography – tandem mass spectrometry method for analysis of 120 veterinary drugs in bovine kidney

“…Sulfonamide and tetracycline recoveries were not significantly influenced by the presence or absence of formic acid. Given the importance of b-lactams in monitoring schemes, [1] the absence of formic acid …”

“…Current monitoring process and plans for improvement have been described elsewhere. [1] Methods for residue monitoring have generally evolved over the years from single-analyte methods, to multi-analyte methods. In recent years, multi-class multi-residue methods (MMMs) have been introduced to further increase monitoring efficiency.…”

“…Typically, such methods utilize liquid chromatography-tandem mass spectrometry (LC-MS/MS) due to the exceptional selectivity and sensitivity of the modern instrumentation. For example, LC-MS/MS has been demonstrated to perform well in the analysis of 62 drugs in bovine kidney, [1] 27 drug analytes from 5 classes in milk, [2] 41 drugs from 7 classes in eggs, [3] and 24-53 analytes from 4-8 classes in animal tissues. [4][5][6][7] LC-MS/MS is capable only of monitoring targeted analytes, and a few newer methods have been reported for even larger numbers of targeted analytes (≥100) from 9-12 classes in milk or animal tissues coupling LC to time of flight mass spectrometry (TOF-MS), or ultra high performance liquid chromatography (UHPLC) with high resolution MS, [8][9][10] with the added potential to find non-targeted chemical contaminants.…”