Electrostacking online sample pre-concentration capillary electrophoresis with amperometric detection for beta2-agonists in human urine

A simple, rapid, sensitive and specific method using capillary electrophoresis (CE) coupled with electrostacking and amperometric detection (AD) has been developed for the simultaneous separation and determination of clenbuterol (CLB), terbutaline (TER), salbutamol (SAL) and formoterol (FMT). In this paper, the CE separation and AD conditions were investigated in detail. The optimum conditions were: pH 8.6 Na(2)B(4)O(7)-H(3)BO(3) buffer solution (20.0 mmol/L), 9 kV for the separation voltage, and 1000 mV (versus Ag/AgCl) for the detection potential. When the sample which was dissolved in 70% ACN-water mixture solution was injected 60 s by 15 kV electrokinetic injection, the best stacking effects was obtained. Under the optimum conditions, the enhancement factors of these beta(2)-agonists had been greatly improved more than 5500-fold compared with the conventional electrokinetic injection. And then an excellent linear response was obtained with LODs (S/N = 3) of 0.098, 0.024, 0.063 and 0.920 pmol/L for CLB, TER, SAL and FMT in urine, respectively. The precision was determined in both intra-day (n = 5) and inter-day (n = 15) assays, and the RSDs were not more than 2.1 and 3.4% for migration time and peak current, respectively. The proposed method has been applied to analyze human urine samples successfully.