We developed a new non-derivatization analytical method for the determination of galactose in the diagnosis of galactosemia by high-performance anion-exchange chromatography (HPAEC)-pulsed amperometric detection (PAD). With an anion-exchange column, the analytes were separated efficiently using 3mM NaOH containing 1mM NaOAc, and 200mM NaOH was added for post-column reagent. The limit of detection (S/N=3) and limit of quantification (S/N=10) for galactose were 25ng/mL and 83ng/mL, respectively. Linear dynamic range was from 4.67mg/dL to 53.46mg/dL (r(2)=0.9999). The mean recovery of galactose for intra-, inter-day assays were found to be of satisfactory results (98.14-101.42%).