Increased national interest in biofuel production has motivated the need for higher throughput analytical methods to facilitate research in pretreatment, enzyme development, and fermentation. Analysis of biomass hydrolysate liquor has applications in all of those areas. Current NREL procedures use an HPLC-based method for the analysis of organic acids, furans, and alcohols in hydrolysate liquor with a run time per sample of over 55 minutes. This research describes a new HPLC method that maintains the accuracy and precision of the existing procedure but reduces the analysis time to less than 10 minutes per sample.