A derivative spectrofluorimetric method for simultaneous determination of enrofloxacin (EFX) and ciprofloxacin (CFX) in cow milk was developed. The sample preparation was based on rotating disk sorptive extraction (RDSE) using Oasis HLB® as extraction phase and followed by desorption of the analytes with methanol. The final extracts were evaluated by derivative spectrofluorimetry.          An analyte-free cow milk was used as blank sample which was spiked with a known amount of analytes for the study of variables. The sample was treated with trichloroacetic acid in order to precipitate proteins. Furthermore, the extraction was carried out in the presence of Mc Ilvaine-EDTA 0.3M buffer to avoid any type of interference from the calcium ion. The variables involved in the RDSE process were pH and extraction time.          Spectral variables were also optimized and analytical signals were evaluated at 417nm and 438nm for EFX and CFX, respectively.          The limit of detection (LOD) and limit of quantification (LOQ) were 2.98 - 9.04 µg · L-1 and 2.56 - 7.75 µg · L-1, for EFX and CFX respectively. The precision levels, expressed as relative standard deviation, for EFX and CFX were 3.2% and 3.0%, respectively. The recoveries were between 98.8% to 100.5%.          Commercial milks were analyzed and both antibiotics shown concentrations below to LOD in the batches analyzed. Finally, the proposed method offers advantages in terms of simplicity, efficiency and cost. Besides being friendly with the environment.