Vortex-assisted liquid–liquid extraction combined with field-amplified sample injection and sweeping micellar electrokinetic chromatography for improved determination of β-blockers in human urine

•The first report of VALLE-FASI-sweeping-MEKC for two beta blockers analysis in urine.•Improving sensitivity via combination of electrophoretic and chromatographic effects.•The electrophoretic and chromatographic pre-concentration parameters were optimized.•Sensitivity enhancement factors of two-step stacking MEKC was 8725-14,074.

A new micellar electrokinetic chromatography (MEKC) method was developed and validated for the analysis of carvedilol and propranolol in human urine samples. In this study, vortex-assisted liquid–liquid extraction (VALLE) coupled with field-amplified sample injection and sweeping was employed for biological sample clean-up and sensitivity enhancement in MEKC. After VALLE, the urine samples were analyzed by MEKC. Tris-phosphate buffer (60 mmol L−1, pH 2.0) containing 40% (v/v) methanol was first filled into an uncoated fused-silica capillary (56 cm, 50 µm i.d.). The pretreated urine sample was loaded by electrokinetic injection (10 kV, 250 s). The stacking and separation were performed using Tris-phosphate buffer (30 mmol L−1, pH 3.0) containing 30% (v/v) methanol and 50 mmol L−1 sodium dodecyl sulfate (SDS) at −25 kV. Detection was carried out at 195 and 214 nm for carvedilol and propranolol, respectively. The suggested method is linear (r2≥0.997) over a dynamic range of 0.005–1 µg mL−1 in urine. The intra- and inter-day relative standard deviation and relative error values of the method were below 20%, which shows good precision and accuracy. Finally, this method was successfully applied to the analysis of real urine samples.

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