Determination of monoamines in urine by capillary electrophoresis with field-amplified sample stacking and...

A simple, rapid and low-cost method using capillary electrophoresis coupled with field-amplified sample stacking and electrochemical detection was developed for the separation and determination of monoamines. In this present work, a systematic study of the parameters (pH value and concentration of electrophoretic buffer, composition of sample solvent, injection voltage and time) affecting separation and on-line concentration of monoamines has been performed enabling the detection sensitivity of these monoamines to be improved by 5,000 times compared with the conventional electrokinetic injection. This developed method was applied to the direct analysis of these monoamines in human urine without off-line sample preconcentration. Due to the requirement for urine dilution to minimize the detrimental effects of high salt on analyte stacking, the real sensitivity improvement is about 50-fold when applying the optimized method to urine samples. In order to quantitate these monoamines accurately, internal standard calibration curves were constructed with standard monoamines in presence of salt with similar concentration as in human urine. In the method validation, the calibration curves were linear over a range of 1.0 x 10(-9) to 2.5 x 10(-8) mol/L for each monoamine and the limits of detection (signal to noise ratio of 3) for these monoamines were in the sub-nmol/L concentration range (6.0 x 10(-10) mol/L).