ICH GUIDELINE PRACTICE: A VALIDATEDPOLARIONICMODE CHIRAL HPLC METHOD DEVELOPMENTANDITSAPPLICATION FOR THE DETERMINATIONOFSAXAGLIPTIN STEREOISOMERS INBULKANDPHARMACEUTICAL FORMULATIONS

SAXAGLIPTIN (SAX) is a commonly used and orally active medicinal ingredient for the therapy ofnon-insulin- dependent diabetes. Three out of all the stereo isomers were generated in minor quantities as impurities duringthesynthesis of SAX, lowering the drug's purity. The primary goal of this work was to investigate several chiral stationary phases (CSPs) in order to identify the phase providing the most efficient and reliable separation of thestereoisomers. Subsequently, chromatographic requirements were adjusted, and the developed technique wascomprehensively confirmed using multiple standard validation parameters. CH3OH:CH3COOH:(C2H5)3N(100:0.1:0.1, v/v/v/) . Compared to Chiralpak-IG, the Chirobiotic V2 column effectively separated the SAXstereoisomers with good resolution in Polar Ionic Mode. The validated chiral HPLC method showed excellent selectivity with resolution >1.95 and tailing factor <1.35 for all analytes. The validated analytical procedureshowed outstanding sensitivity, with limits of detection ranging from 0.32 to 0.68 µg/mL and limits of quantification spanning 1.09 to 2.13 µg/mL for DIA-1, DIA-2, ENA, and SAX. The proposed analytical strategy demonstrated strong selectivity, precision, accuracy, linearity, and robustness for the reliable identificationand quantification of SAX stereoisomers, according to ICH guidelines. The described approach has been usedtodetermine the stereoisomers of SAX bulk pharmaceuticals and pharmaceutical formulations with great success.