A new pressure-assisted capillary electrochromatography coupled with electrospray ionization-mass spectrometry method using a silica-based monolithic column as separation media was developed for the analysis of beta(2)-agonists in human urine. Experimental conditions including the mobile phase, separation voltage, assisted pressure, and sheath liquid were optimized for the analysis: mobile phase composed of 82% (v/v) ACN and 18% (v/v) 20mmol/L ammonium acetate (pH 6.0); separation voltage 25kV; assisted pressure 2 bar; and the sheath liquid consisting of 7.5mmol/L acetic acid in isopropanol/water 50/50% (v/v) that was delivered at a flow rate of 3.0microL/min. Six beta(2)-agonists were separated within 12.5min with LODs (defined as S/N=3) in the range of 0.25-2.0ng/mL. The absolute LODs of the developed method for analyzing six beta(2)-agonists ranged from 5.75 to 46.0fg. Method repeatability of run-to-run and column-to-column was satisfactory. The recovery obtained from the analysis of spiked urine samples was between 88.2% and 106% with RSDs lower than 6.68%. The method was successfully applied to the analysis of real urine sample from volunteers.
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