Enzymatic hydrolysis is one of the main steps in the processing of bioethanol from lignocellulosic raw materials. However, complete understanding of the underlying phenomena is still under development. Hence, this study has focused on validation of the inhibition effects in the cellulosic biomass hydrolysis employing a dynamic mathematical model. A systematic framework for parameter estimation is used for model validation, which helps overcome the problem of parameter correlation. Data sets obtained from carefully designed enzymatic cellulose and cellobiose hydrolysis experiments, were used for parameter estimation (calibration) and validation purposes. The model predictions using calibrated parameters have shown good agreement with the validation data sets, which provides credibility to the model structure and the parameter values.