The use of veterinary drugs on animals, including bees, might result in the accumulation of residues in food items like honey. Because of its excellent sensitivity and selectivity, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is widely accepted as the most effective technology for identifying and assaying these residues in complex matrices. However, obstacles remain for the concurrent extraction and purification of residues with diverse physicochemical characteristics. This study aims to develop and validate a method for identifying and confirming antibiotic residues in honey, in accordance with European Regulation 2021/808/EC, which sets the requirements for analytical methods and the interpretation of the results. The validation findings reveal that the technique is particular, linear (with regression coefficients around 99%), fair (recovery rate between -30% and +20%) and faithful (consistent coefficients of variation). The decision limits (CCa) reached are less than the minimum performance requirements (MMPR). No residue was detected in the white samples, supporting the method’s specificity. Finally, the LC-MS/MS technique for detecting antibiotic residues in honey was verified and it meets Regulation 2021/808/EC requirements, ensuring trustworthy and accurate findings.