The impact of a realistic error in the column hold-up time on the determination of the adsorption isotherm model was systematically investigated. Frontal analysis and the inverse method were used for the accurate determination of the adsorption isotherm. The true retention times of the breakthrough curves were used with a known hold-up time as reference. The adsorption isotherms were calculated using the same procedure that is used for real experimental adsorption isotherms, where the true hold-up time is unknown. The raw data were analyzed with calculations of adsorption energy distributions (AEDs), Scatchard plots, fitting to different rival adsorption models and finally their ability to predict true profiles. The results show that for a true Langmuir and bi-Langmuir model with an underestimated hold-up time the error may lead to a more heterogeneous model and for overestimated cases false adsorption processes like multi-layer adsorption or solute-solute interaction are assumed. The Scatchard plots for data obtained using a Langmuir adsorption isotherm are nonlinear and the AEDs show clear deviations from Langmuir behavior already at small deviations from the true hold-up time at a moderate surface coverage. The inverse method confirms the result that was obtained from the frontal analysis procedure.