Quantitative analysis of clonidine and ephedrine by a microfluidic system: On-chip electromembrane extraction followed by high performance liquid chromatography

In this work, a microfluidic device was developed for on-chip electromembrane extraction of trace amounts of ephedrine (EPH) and clonidine (CLO) in human urine and plasma samples followed by HPLC-UV analysis. Two polymethylmethacrylate plates were used as substrates and a microchannel was carved in each plate. The microchannel channel on the underneath plate provided the flow pass of the sample solution and the one on the upper plate dedicated to a compartment for the stagnant acceptor phase. A piece of polypropylene sheet was impregnated by an organic solvent and mounted between the two parts of the chip device. An electrical field, across the porous sheet, was created by two embedded platinum electrodes placed in the bottom of the channels which were connected to a power supply. The analytes were converted to their ionized form, passed through the supported liquid membrane, and then extracted into the acceptor phase by the applied voltage. All the effective parameters including the type of the SLM, the SLM composition, pH of donor and acceptor phases, and the quantity of the applied voltage were evaluated and optimized. Several organic solvents were evaluated as the SLM to assess the effect of SLM composition. Other parameters were optimized by a central composite design. Under the optimal conditions of voltage of 74 V, flow rate of 28 mu L min(-1), 100 and 20 mM HCl as acceptor and donor phase composition, respectively, the calibration curves were plotted for both analytes. The limits of detection were less than 7.0 and 11 mu g L-1 in urine and plasma, respectively. The linear dynamic ranges were within the range of 10-450 and 25-500 mu g L-1 (r(2) (>) 0.9969) for CLO, and within the range of 20-450 and 30-500 mu g L-1 (r(2) (>) 0.9907) for EPH in urine and plasma, respectively. To examine the capability of the method, real biological samples were analyzed. The results represented a high accuracy in the quantitative" analysis of the analytes with relative recoveries within the range of 94.6-105.2%" and acceptable repeatability with relative standard deviations lower than 5.1%.