The paper establishes a new HPLC method for measurement of the total content of gluconic acid (GA) in wine using a pulsed amperometric detector equipped with a gold electrode cell and an anion exchange column. A NaOH solution was used at a flow rate of 1.5 mL/min as a mobile phase under isocratic conditions. Analysis took 30 minutes. The linearity of the method was shown up to 500 mg/L, with a LOQ of 2.0 mg/L (10σ). Precision (as RSD%) was under 2.5 % in the usual range of GA concentration in natural wines. Accuracy was checked in comparison with the usual enzymatic method.