Pre-column Derivatization and Separation of Diastereomeric-Derivatives of Racemic Mexiletine and Confirmation of Elution Order and Molecular Configuration

Abstract

Present study describes the synthesis of cyanuric chloride based four active chiral reagents (ACRs) and their application in the enantiomeric separation of (RS)-mexiletine. Herein, four cyanuric chloride-based ACRs were prepared by introducing L-proline derivatives under nucleophilic substitution reaction. The synthesized ACRs were characterized by different spectroscopic techniques. Racemic mexiletine hydrochloride was used for the enantio-recognition study. All the four ACRs were used to convert (RS)-mexiletine into related diastereomeric derivatives and then separated on the C18-column of RP-HPLC. The different parameters such as sample amount, the concentration of mobile phase, organic modifier and pump pressure were varied to optimize separation conditions. The energy-minimized structures of synthesized diasteromeric derivatives (DDs) were developed using DFT calculations. The validation study was conducted for the developed method and correlation-coefficient, calibration range, LOD and LOQ calculated. The stability and recovery were calculated by inter and intraday assay.