The aim of this study was to find the best design that is suitable for optimizing the recovery of the representatives of the 2nd, 3rd and 4th generation of fluoroquinolones. The following designs were applied: Central Composite Design, Box−Behnken Design and Doehlert Design. The recovery, which was a dependent variable, was estimated for liquid−liquid extraction. The time of shaking, pH, and the volume of the extracting agent (dichloromethane) were the independent variables. All results underwent the statistical analysis (ANOVA), which indicated Central Composite Design as the best model for evaluation of the recovery. For each analyte, an equation was generated that enabled to estimate the theoretical value for the applied conditions. The graphs for these equations were provided by the Response Surface Methodology. The statistical analysis also estimated the most significant factors that have an impact on the liquid−liquid extraction, which occurred to be pH for ciprofloxacin and moxifloxacin and the volume of an extracting solvent for levofloxacin.