This study investigated relative magnitude of hydrolysis and ammonification by separate analysis of ammonia release and nitrification mechanisms. A peptone mixture was used as substrate in two parallel experiments seeded with nitrifying biomass conducted with and without nitrification inhibitor. Results were evaluated by means of model analysis of the ammonia and the oxygen uptake rate (OUR) profiles. A dual hydrolysis mechanism with maximum rate coefficients of 6.3 and 0.5/day characterized the peptone mixture and a kinetic balance was established for the ammonia release mechanism with a corresponding ammonification rate of 0.08 m(3)/g COD day. The experiments also showed a low soluble ammonia nitrogen generation that was rapidly depleted, confirming the existence of ammonification. These rate coefficients were verified using model calibration of the OUR profile related to simultaneous carbon removal and nitrification. Results indicated that ammonification would not be rate limiting for wastewaters such as domestic sewage, with lower hydrolysis kinetics. (C) 2012 Elsevier Ltd. All rights reserved.