Enantioselective Separation of Imeglimin API on Novel Immobilized Cellulose-tris (3-chloro-5- methyl phenyl carbamate by HPLC and Preparative purification by SFC Techniques

Abstract

The present study describes the enantiomeric separation of Imeglimin hydrochloride using validated high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) methods. The chiral recognition capabilities of cellulose- and amylose-based chiral stationary phases (CSPs) were systematically evaluated under both HPLC and SFC conditions. Chromatographic performance was assessed by comparing retention time, enantioselectivity, resolution, column efficiency, and peak symmetry under optimized separation conditions. The influence of organic modifiers and column temperature on enantiomeric separation was evaluated during method development. Compared with HPLC, selectivity in SFC was less affected by variations in the type and composition of alcohol modifiers (ethanol and isopropyl alcohol). The observed retention and enantioresolution suggest that hydrogen-bonding interactions play a dominant role in the chiral recognition mechanism, enabling efficient separations with shorter analysis times. Furthermore, the SFC method offers potential for preparative-scale applications owing to the facile removal and recovery of the mobile phase compared with conventional HPLC. Despite the advantages of SFC, the developed HPLC method was selected for validation owing to its simplicity, use of conventional LC instrumentation, and suitability for routine quality control applications under isocratic conditions. The method was successfully validated in accordance with ICH guidelines and applied to the analysis of multiple batches of Imeglimin hydrochloride bulk drug. Furthermore, the applicability of the newly introduced CSP was demonstrated with several active pharmaceutical ingredients, providing excellent enantiomeric resolution and superior performance compared with previously reported chiral HPLC methods. Keywords: Enantiomeric separations; CHIRALPAK IK; Imeglimin; HPLC; SFC.