In the contemporary landscape, polysaccharide-based chiral stationary phases have garnered significant popularity for several reasons: (i) their broad application range, (ii) the availability of diverse chemistry options in both coated and immobilized forms, and (iii) their substantial loading capacity, advantageous for preparative-scale operations. Undoubtedly, chiral separations continue to be a focal point, especially within the pharmaceutical sector, where supercritical fluid chromatography is swiftly gaining traction. However, its utilization presents greater complexity compared to high-performance liquid chromatography. The provided examples serve to elucidate the development of analytical-scale chiral separation methods and the impact of each operational parameter, such as flow rate, outlet pressure, and temperature variations, on chiral separation.