The activated sludge models (ASMs) commonly used by the InternationalWater Association (IWA) task group are based on chemical oxygen demand (COD) fractionations. However, the proper evaluation of COD fractions, which is crucial for modelling and especially oxygen uptake rate (OUR) predictions, is still under debate. The biodegradation of particulate COD is initiated by the hydrolysis process,which is an integral part of anASM. This concept has remained in use for over 30 years. The aim of this study was to verify an alternative, more complex, modified (Activated Sludge Model No 2d) ASM2d for modelling the OUR variations and novel procedure for the estimation of a particulate COD fraction through the implementation of the GPS-X software (Hydromantis Environmental Software Solutions, Inc., Hamilton, ON, Canada) in advanced computer simulations. In comparison to the original ASM2d, the modified model more accurately predicted the OUR behavior of real settled wastewater (SWW) samples and SWWafter coagulation-flocculation (C-F). The mean absolute relative deviations (MARDs) in OUR were 11.3-29.5% and 18.9-45.8% (original ASM2d) vs. 9.7-15.8% and 11.8-30.3% (modified ASM2d) for the SWWand the C-F samples, respectively. Moreover, the impact of the COD fraction forms and molecules size on the hydrolysis process rate was developed by integrated OUR batch tests in activated sludge modelling. © 2020 by the authors.