Pre-column Derivatization¸ Elution Order, Molecular Configuration and Green Chromatographic Separation of Diastereomeric Derivatives of β-Amino Alcohols

A green chromatographic method was developed for the indirect enantio-separation of racemic amino alcohols by introducing a micellar mobile phase for the HPLC. Here, cyanuric chloride-based activated chiral reagents were prepared, characterized, and used to derivatize racemic amino alcohols into diastereomeric derivatives. The chromatographic separation of the prepared diastereomeric derivatives was achieved on the C18 column of the RP-HPLC using a surfactant-based (Brij-35 + SDS) aqueous mobile phase (as an alternative to organic solvents). The impact of the varying concentration of surfactants in the mobile phase and pH of buffer on the chromatographic separation was optimized. Additionally, the DFT calculations were performed, and the absolute configurations and elution orders of the diastereomeric derivatives were investigated. Developed method’s robustness and selectivity was optimized by performing validation studies (LOD = 0.286 ng/mL; LOQ = 0.858 ng/mL). Also, the green assessment score was calculated for the developed method (84 scores; an excellent green method).