Simultaneous separation and quantification of pomalidomide chiral impurity using reversed phase–ultra-high-performance liquid chromatography method

Objectives: This study develops and validates an RP-UHPLC method for simultaneously separating and quantifying the R(+) and S(-) enantiomers of Pomalidomide, a 2 nd generation immunomodulatory drug, in API using an orthogonal experimental design approach. Materials & Methods: Chromatographic conditions involve an Ultisil Cellu-JR column, a flow rate of 0.8 mL/min, an injection volume of 10 µL, and a 15-minute run time at ambient temperature. Peak purity analysis, facilitated by OpenLab LC software, ensures the reliability of separation by evaluating UV spectra similarity within the 170–700 nm range. Results: The method successfully achieves Pomalidomide enantiomeric separation in the isocratic mode, with retention times of 7.507 minutes for the enantiomer-I and 8.943 minutes for the enantiomer-II. Conclusion: The study underscores the need for diverse chromatographic methods to overcome the challenges posed by Pomalidomide’s chiral nature. The results provide valuable insights into optimizing chromatographic techniques for the analysis of chiral impurities, contributing to the advancement of pharmaceutical quality control and research.