Determination of Baclofen Residue in Muscle,Liver, Kidney and Fat of Swine by Liquid Chromatography-Tandem Mass Spectrometry

Baclofen was illegally used in veterinary clinical medicine as a growth-promoting agent. To date, few methods have been developed for the monitoring of baclofen in animal tissues. In this study, a sensitive and efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to identify and quantify baclofen in the muscle, liver, kidney, and fat of swine was developed and validated. Baclofen was extracted from tissues with ammonium acetate buffer (pH 5.2) and isolated with isopropyl-ethyl acetate (4:6, v/v). Then, a solid phase extraction using MCX cartridge was used to clean up the extracts. The elution was evaporated to dryness and reconstituted with water/methanol (90:10 v/v). All samples were determined by LC-MS/MS system through positive ionization in a multiple reaction monitoring (MRM) mode. The proposed method was validated by evaluation of specificity, linearity, recovery, accuracy, precision, LOD, and LOQ values according to Commission Decision 2002/657/EC. Estimated limit of quantification for baclofen in the muscle, liver, kidney, and fat of this method was 1.00 μg/kg, respectively. The mean intra- and inter-day assay accuracies fell within a range 88.5–93.9% and 86.2–93.2%, respectively. The mean intra- and inter-day precisions were 1.78 and 4.95% (RSD < 15%), respectively. The proposed method has proved to be suitable for accurate quantitative determination of baclofen for residue analysis.