Abstract The objective of this study was to evaluate NBPT, zeolite and humic acid in different concentrations as inhibitors of the activity of the enzyme urease. The activity of the urease enzyme was quantified by the methods proposed by May and Douglas (1976) and Witte and Medina-Escobar (2001). For this reason, two experiments were conducted in a completely randomized design in split plots, in which the incubation periods are the plots and the fertilizers with the inhibitors at the different concentrations are the subplots with three replications. Fifteen fertilizers based on NBPT-coated urea, humic acid and zeolite were used in different concentrations. The two methods of analysis used in the experiment confirmed the efficiency of NBPT in inhibiting the enzymatic action even at low concentration, with 0.02% (m/m) being sufficient to inhibit urease, with no difference between inhibitor concentrations. The addition of humic acid reduced the activity of urease after 36 hours of reaction when the activity of the enzyme was evaluated by the method proposed by May and Douglas (1976). Zeolite did not influence the activity of the urease enzyme when analyzed by the May and Douglas method (1976).