Stereospecific high-performance liquid chromatographic assay of zopiclone in human plasma

A high-performance liquid chromatographic (HPLC) assay for the analysis of the enantiomers of zopiclone (ZPC), a cyclopyrrolone hypnotic, in plasma was developed. Following the addition of chlordiazepoxide as internal standard (I.S.), plasma containing the ZPC enantiomers and I.S. was extracted by liquid-liquid extraction at an alkaline pH. After evaporation of the organic layer, the drug and I.S. were reconstituted in ethanol-hexane (80:20, v/v) and injected onto the HPLC column. The enantiomers were separated at ambient temperature on a 25-cm Chiralcel OD-H column with ethanol-hexane (60:40, v/v) as the mobile phase pumped at a flow-rate of 0.6 ml/min. The enantiomers of ZPC were quantified by fluorescence detection with excitation and emission wavelengths of 300 and 470 nm, respectively. The assay described allows for the direct quantitation of ZPC without pre-column derivatization, and is suitable for clinical studies of ZPC in humans after administration of therapeutic doses.