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Introduction: Mannich base is the result of the three-part chemical reaction known as the Mannich reaction, which includes the amino alkylation of an acidic proton adjacent to a carbonyl group by formaldehyde and either a primary or secondary amine. Method: In a 1L flask, dissolve 28.2 g (0.3 mol) of 2-aminopyridine, and 400 mL of ethanol, then add 32.6 g (0.3 mol) of chloroacetone and put 30 drops of acetic acid. The organic phases are combined and dried with Magnesium sulfate. After solvent evaporation, we have recovered our product which is a 2- methylimidazo [1, 2-a] pyridine product. Result: A C-18 bonded stationary phase provided one significant peak for all of the compounds under examination providing solid proof of the compound purity. This was used to demonstrate the purity of the compounds. Then, the potential enantiomers of these chiral compounds were obtained using the four CSPs. Conclusion: In this study, several novel Mannich bases were prepared. These bases had rendement ranging from 76 to 94.73, and their chiral separation was discussed utilizing four polysaccharide-based CSPs: ChiralpakAS, Chiralcel OD, Chirapak®AS-3R, and Chiralcel OJ.