Switchgrass is a potential source of renewable biomass for conversion to liquid biofuels. Efficient conversion requires effective strategies for pretreatment and enzymatic saccharification to produce fermentable sugars. Standard analysis of fermentation liquids includes detection of monosaccharides and ethanol to determine efficiency of conversion. Larger components, specifically oligosaccharides, are typically not measured due to the structural complexity of the products; however, as oligosaccharides they represent carbon available in biomass that is not converted to liquid fuels. In this study, ammonia-pretreated switchgrass was enzymatically depolymerized either independently or under simultaneous saccharification and fermentation conditions. Residual oligosaccharides were reducing end-labeled followed by hydrophilic interaction liquid chromatography mass spectrometry/mass spectrometry analysis. These data reveal 20 oligosaccharide peaks with distinct retention times and tandem mass spectrometry fragmentation patterns representing 13 different oligosaccharide compositions. All measured compositions were smaller than a chain length of six and were neither linear xylooligosaccharides nor modified with phenolic esters. This work represents a robust method to monitor and identify unhydrolyzed oligosaccharides from fermentations, thereby permitting the screening of targeted enzymatic activities to promote the complete hydrolysis of xylan. Published by Elsevier Ltd.