The most accurate analysis method for sugars in biomass, based on gas chromatography, requires a time consuming and laborious sample derivatation to trimethylsilanes or alditol acetates. In comparison, sample preparations for sugar analysis by liquid chromatography are simple water dilutions. However, HPLC methods either require long analysis times, use of expensive solvents, or do not give good resolution of sugars. A gradient method developed previously using a Dionex PA-1 column and pulsed-amperometric detection was modified to reduce analysis time from 75 to less than 40 min and provide good resolution of arabinose, rhamnose, galactose, xylose, glucose, fructose, sucrose, cellobiose, and galacturonic acid in both standards and hydrolyzed citrus waste biomass.