Chiral Separation and Thermodynamic Investigation of WCK 3023: A Novel Oxazolidinone Antibacterial Agent, Application to Pre‐clinical Pharmacokinetic Study

: A chiral liquid chromatographic method was developed and validated for the quantification of R-enantiomer impurity (RE) in WCK 3023 (S-enantiomer), a new drug substance. The separation was achieved on Chiralpak IA (amylose-based immobilized chiral stationary phase), using a mobile phase consisting of n-hexane-ethanol-trifluoroacetic acid (70:30:0.2, v/v/v) at a flow rate of 1.0 mL/min. The method was extensively validated for the quantification of RE in WCK 3023 and proved to be robust. For RE the detector response was linear over the concentration range of 0.11-5 μg/mL. The limit of quantitation and limit of detection for RE were 0.11 and 0.04 μg/mL respectively. Average recovery of the RE was in the range of 98.11-99.55%. The developed method was specific, sensitive, precise and accurate for quantitative determination of RE in WCK 3023. The impact of thermodynamic parameters on the chiral separation was evaluated. The method was employed for controlling the enantiomeric impurity in the lots of WCK 3023 used for pre-clinical studies. The method was successfully applied to evaluate the possible conversion of WCK 3023 to RE in rat serum samples during pre-clinical pharmacokinetic studies.