A new liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed to confirm chloramphenicol (CAP) residues in foods of animal origin and in urine samples, which were earlier found positive under the screening analysis, performed by competitive enzyme-linked immunoassay (ELISA) technique. The developed LC-MS/MS method was applied to four non-compliant samples from 2008 to 2012; giving concentrations of CAP residues from 1.18 to 3.68 μg kg−1. All samples, qualified positive by ELISA, were confirmed with the LC-MS/MS technique and found to be non-compliant. The effectiveness of the confirmatory method was proven by participating in a successful proficiency test in year 2010. Both LC-MS/MS and ELISA methods were validated according to the European Union 2002/657/EC decision. The decision limit of the confirmatory method was determined as 0.02 μg kg−1 for CAP in each validated matrix, while the detection capability of the screening test was 0.15 μg kg−1.