Abstract The aim of this study was to develop a simple high-performance liquid chromatography (HPLC) with UV detection method, for the determination of four sulfonamides in chicken muscle tissue. The sulfonamides were extracted with acetonitrile, acetone and dichloromethane. Separation was carried out on an C18 analytical column, using as mobile phase a mixture of 6‰ di-sodium hydrogen phosphate and methanol. The analytes were detected by UV, in one run. Calibration curves were linear with very good correlation coefficients for concentration ranging from 30μg kg-1 to 150μg kg-1. The limits of detection (LOD) for sulfonamides ranged from 6.5 to 0.14 μg kg-1. The recovery for spiked chicken muscle with 50-150 μg kg-1 ranged more than 70%. The relative standard deviations (RSDs) of the sulfonamides for six measurements at 50 μg kg-1, 100 μg kg-1 and 150 μg kg-1 were less then 15%. These parameters met the EU criteria for method validation. The results were confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using multiple reacting monitoring, as operating mode. Confirmation require the retention times of the analytes to be within ±2.5% of the retention times of the standards, presence of the parent ion and two characteristic fragment ions (product ions) per analyte, as well as the relative ion abundance ratios of the fragment ions shall correspond to ratios obtained for the standards, within permitted limits . The transition of two common product ions at m/z 155.7 and 107.5 were monitored for all sulfonamides. Each of the analytes, in all tested samples met the confirmation criteria. Thus, it was demonstrated the applicability of the HPLC-UV method for routine analysis of chicken muscle tissue.