Determination of thiols in urinary sample by capillary-column liquid chromatography with amperometric detection at a carbon electrode

The fabrication and application of an end-column amperometric detection (AD) system with a carbon electrode for capillary-column liquid chromatography (CLC) were described. This new amperometric detector showed good sensitivity and stability in detecting the thiocompounds and other analytes with CLC. In order to obtain the better separation and detection performance for analytes, several operational parameters had been investigated: the working potentials, pH and flow rate. Under the optimum conditions, the method could effectively separate and determine cysteine (Cys), glutathione (GSH), dopamine (DA) and 6-thiopurine (6-TP). Good repeatability for retention time was obtained with a relative standard deviation (R.S.D.) value of 0.5%. The linear range covered over three orders of magnitude and the limits of detection were 8 fmol for cysteine, 20 fmol for glutathione, 8 fmol for dopamine and 20 fmol for 6-thiopurine. This method was successfully applied in determination of urinary sample with characteristics of simplicity, high sensitivity and good repeatability.