Methodology for the analysis of etomidate and its enantiomer by HPLC

This study presents a comprehensive methodology for the analysis of etomidate and its enantiomer using high-performance liquid chromatography (HPLC). Etomidate, a widely used anesthetic agent, exists as a racemic mixture, and its enantiomers may exhibit different pharmacological properties. Therefore, the separation and quantification of both the drug and its enantiomer are critical for understanding their respective therapeutic effects and safety profiles. The proposed HPLC method utilizes a chiral stationary phase to achieve optimal resolution of etomidate and its enantiomer. The mobile phase consists of a combination of methanol and phosphate buffer, adjusted to pH 7.0, providing a suitable environment for the separation. The flow rate is set at 1.0 mL/min, and detection is conducted at a wavelength of 240 nm, which corresponds to the UV absorbance of etomidate. Calibration curves for both etomidate and its enantiomer demonstrate linearity in the range of 0.1 to 10 μg/mL, with correlation coefficients exceeding 0.99. The method exhibits excellent precision and accuracy, with intra-day and inter-day variability within acceptable limits. Limit of detection (LOD) and limit of quantification (LOQ) are established, ensuring the method’s sensitivity for trace analysis. The validated HPLC method has been successfully applied to the analysis of etomidate in plasma samples, highlighting its potential for clinical pharmacokinetic studies. This methodology not only enhances the understanding of etomidate’s therapeutic effects but also paves the way for future studies on the pharmacological significance of its enantiomeric forms.