The study was focused on optimization of methods for the determination of chemotherapeutics from the group of sulfonamides (sulfadiazine, sulfathiazole, sulfamethoxazole, sulfadoxin) in honey by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Three working processes were selected for the optimization: water, acid hydrolysis 1, acid hydrolysis 2. The aim of the study was to determine a suitable methodology in which extraction of selected sulfonamides would be acceptable in terms of their physico-chemical properties. The model study was performed on a set of twelve parallel determinations at concentrations of 0.5, 1 and 5 μg/L for each procedure. The gained results indicated following analytical recoveries: from 79 % to 121 % (water), from 88 % to 127 % (acid hydrolysis 1) and from 61 % to 95 % (acid hydrolysis 2). The repeatability, expressed as relative standard deviation (RSD) was from 7.73 % to 29.60 % (water), from 4.37 % to 14.74 % (acid hydrolysis 1) and from 13.96 % to 31.58 % (acid hydrolysis 2). Regarding the physico-chemical properties of sulfonamides, acid hydrolysis 2 was chosen as a suitable process.
