In this paper, we describe the enantiomeric separation of a chiral alcohol under subcritical fluid chromatography conditions using a 3 μm particle size bonded amylose carbamate stationary phase. Linear and branched alcohols were used as polar modifiers in CO2. The studies with linear alcohols showed a decrease in selectivity factor as the number of carbons in the linear chain increased. For branched alcohols, as the bulk of substituents at the α carbon atom increases the separation factor decreases. Thermodynamic studies showed that in the presence of the alcohols studies, except methanol and ethanol, a positive ΔΔS was observed. Molecular mechanics simulation brought more insights into the mechanism of enantiomeric separation on this stationary phase under subcritical fluid chromatography.