Ultrasensitive analysis of mirtazapine and its metabolites enantiomers in body fluids using ultrasound-enhanced and surfactant-assisted dispersive liquid-liquid microextraction followed by polymer-mediated stacking in capillary electrophoresis

Abstract

A simple, rapid, and sensitive technique for measuring mirtazapine and its metabolites enantiomers in human fluids, such as urine and serum, was developed by applying ultrasound-enhanced and surfactant-assisted dispersive liquid-liquid microextraction (USA-DLLME) integrated with poly(diallyldimethylammonium chloride) (PDDAC)-mediated stacking in capillary electrophoresis (CE). The parameters that affect extraction and stacking performance, such as the extraction volume, surfactant types, surfactant concentrations, salt additives, extraction time, solution pH, and background electrolytes, were comprehensively studied and optimized to achieve optimal detection performance. Under optimal extraction conditions (injection of 120 µL of C₂H₂Cl₄ into 1 mL of a sample solution containing 0.05 mM Brij-35 at pH 10.0) and separation conditions (0.9% PDDAC, 10 mM phosphate, pH 3.0, and 20 mM dimethyl-β-cyclodextrin), on-line CE stacking of mirtazapine-related chiral drugs was achieved by the two strategies: (i) neutral DM-β-CD sweep low concentrations of DL-NaSSA and (ii) DL-NASSA is stacked by the difference in the viscosity between the PDDAC and sample zone. An approximately 2,800-4000-fold improvement in detection sensitivity was revealed for mirtazapine, N-demethylmirtazapine, and 8-hydroxymirtazapine enantiomers. The linear ranges for the quantification of all analyte enantiomers were 1.2-150 nM, with a coefficient of determination higher than 0.99; the relative standard deviations in the migration time and peak areas for six analytes were less than 1.8% and 5.8%, respectively. The proposed system provided the limits of detection (signal-to-noise ratio of 3) of the six analytes as 0.3-0.5 nM. The recovery of the six separated analytes spiked in urine and serum samples was revealed to be 82.7%-109.5% and 91%-112.8%, respectively. This advanced technique with high sensitivity enhancement factors was successfully employed to analyze mirtazapine and its metabolites enantiomers in urine and serum samples with reliability.

Keywords: Body fluids; Capillary electrophoresis; Polymer-mediated e stacking; Psychotropic drugs; Ultrasound an surfactant assisted dispersive liquid liquid microextraction.