A sensitive HPLC-electrochemical detection method was developed for the analysis of gliclazide (GL) in human plasma. After deproteination of 100 microL of plasma by acetonitrile, evaporation, and reconstitution, GL was separated on a C18 column (150 mm x 4.6 mm) by the mobile phase (70 mM disodium tetraborate, pH 7.5, containing 26.5% of acetonitrile). The regression equations were linear (r> 0.9990) over the range of 50 nM to 4.00 microM. The precision and accuracy of intra- and inter-day analysis were less than 5.3 and 0.93% for relative standard deviation and relative error, respectively. The limit of detection for plasma was 10 nM for GL (S/N = 3, 10 microL injection). This newly developed method was applied for monitoring blood levels with one healthy volunteer dosing with a GL tablet.