Determination of Xylazine and 2,6‐Xylidine in Animal Tissues by Liquid Chromatography—Tandem Mass Spectrometry

Abstract Xylazine is a potent α2‐adrenergic agonist used in veterinary medicine for sedation, analgesia, muscle relaxation, and so on. Its residue in animal‐derived food may cause the food safety problem. Moreover, the metabolite 2,6‐xylidine was reported to be a genotoxic and carcinogenic compound. Therefore, it is necessary to develop a high sensitive method for analyzing xylazine and metabolite residue in animal products. Here, we described a LC‐MS/MS method for simultaneous determination of xylazine and 2,6‐xylidine in 4 animal tissues: liver, meat, kidney, and fat. The samples were extracted by acetonitrile, and further clean up by hexane. The analysis was performed on a C18 reversed‐phase column and API 5000 Triple Quadrupole mass spectrometry with positive electrospray ionization interface operating in the multiple‐reaction monitoring mode. For all of the investigated sample matrix, the limit of detection (limit of quantitation) for xylazine and 2,6‐xylidine were 0.06 (0.2) and 1.5 (5) μg/kg, respectively, the recoveries were between 63.5% and 90.8%. The precision was within the range of required criteria for method development. The presented method is sensitive and reproducible, and thus suitable for accurate quantification of xylazine and metabolite residue in animal‐derived food products.