The aim of the study was to determine the degree of an organic substrate consumption in the denitrification process involving a biofilm in four-stages laboratory scale rotating biological contactor (RBC). The discs submergence was 40% of their diameter. Acetic acid, used as external carbon source, was fed to the fourth stage of RBC. Consumption of substrate was observed for 2 and 24 hours. For a shorter period there was the removal of 37.9 ± 1.8 mg N·m^-2, while using 499,9 ± 33.2 mg O₂·m^-2 of organic compounds. The prolongation of the experiment duration guaranteed higher efficiency of denitrification. The ratio of organic substrate used to the amount of nitrogen removed was 13: 1 and 21: 1 for 2 and 24 hours respectively.