Due to a great demand for amylose and cellulose polymeric chromatographic chiral columns, the enantiomeric separation of thiourea derivatives was achieved on the different amylose (Chiralpak-IB) and cellulose chiral (Chiralcel-OJ and Chiralcel-OD-3R) columns with varied chromatographic conditions. The mobile phases used were the different combinations of n-hexane and methanol. The separation and resolution factors for all the enantiomers were in the range of 1.25 to 3.47 and 0.48 to 1.75, respectively. The enantiomeric resolution was obtained within 12 minutes making fast separation. The docking studies confirmed the chiral recognition mechanisms with binding affinities in the range of -4.7 to 5.7 Kcal/mol. The reported compounds have good anti-coagulant activities and may be used as anti-coagulants in the future. Besides, chiral separation is fast and is useful for enantiomeric separation in any laboratory in the world.