Chiral columns based on cellulose tri(3,5-dimethyl phenylcarbamate) (CTDMPC) have been extensively used for enantioseparation by high-performance liquid chromatography (HPLC). The effects of cellulose derived using different amounts of 3,5-dimethyl phenylcarbamate and silica gels prepared by different methods on the chiral separation characteristics have been investigated. Thirteen chiral chromatographic columns were prepared, and their separation capability for sixteen racemate samples was evaluated. The efficiency of enantioselective resolution followed the order trisubstituted cellulose column > disubstituted cellulose column > cellulose column. Refine silica gel and macroporous silica gel showed better performance than did coarse silica gel. The use of macroporous silica gel resulted in low column pressure. Whether the silica gels were derived by aminopropyl could affect the enantioseparations. The thirteen chiral chromatographic columns were complementary for the separation of racemates, especially the cellulose column. This is a comparative study on the preparation of chiral stationary phases for HPLC, and the findings would enable us to design and fabricate chromatographic columns.