Simultaneous extraction and analysis of clozapine and lorazepam from human plasma using dual solvent-stir bar microextraction with different acceptor phases followed by high-performance liquid chromatography ultra-violet detection

Abstract

A new design of dual solvent stir bar microextraction (DSSBME) was developed and combined with HPLC-UV for the simultaneous extraction of clozapine (CLZ) and lorazepam (LRP) from human plasma with different acceptor phases. Two short hollow fibers immobilized with an organic extraction solvent were used as the solvent bars for microextraction of CLZ and LRP from the sample solution. The solvent bars were fixed with a staple pin which served as the stirrer. The target analytes were simultaneously and selectively extracted from the sample solution into their corresponding solvent bar. Extraction parameters such as organic solvent type, pH of the sample solution, the acceptor phase concentration, salt incorporation into the solution, stirring rate, and extraction time were optimized to achieve the best extraction results. Under the optimum conditions (1-undecanol as extraction solvent, pH of sample solution = 9.0, 10% w/v NaCl, concentration of HCl = 10 mM, concentration of NaOH = 100 mM, stirring rate of 1400 rpm and extraction time of 30 min at ambient temperature) the limit of detection for CLZ was 0.4 ng mL⁻¹ and for LRP it was 1.1 ng mL⁻¹. The linear range for CLZ was 1.3–1000.0 ng mL⁻¹ (R² = 0.9991) and for LRP it was 3.6–800.0 ng mL⁻¹ (R² = 0.9993). Extraction recovery and the enrichment factor for CLZ were 95.4% and 343 and for LRP they were 74.3% and 263, respectively. Finally, the method developed was successfully applied for the simultaneous determination of CLZ and LRP in human plasma samples.