A semipreparative column packed with microcrystalline cellulose triacetate (MCTA) was used to separate ketamine enantiomers by high-performance liquid chromatography (HPLC). The effect of temperature on the kinetics of mass transfer and equilibrium constants were evaluated by the moment analysis at 303, 313, and 323 K. Total porosity, bed porosity, and equilibrium constants were measured by the first moment. Pore diffusion and axial dispersion coefficients were evaluated by the second moment. It was observed that band broadening decreased with increasing temperature. The equilibrium constants were found to be greater than unity and decreasing with increasing temperature. The pore diffusion coefficients increased with increasing temperature and the main contribution to the band broadening was attributed to low mass transfer kinetics. The pore diffusion coefficients controlled the mass-transfer process in MCTA column. These results may be used to the determination of operating conditions and computational simulation of a chromatographic separation in simulated moving bed unity.