Perdeuterated pyridinium chloride-DMSO-d(6) is an effective solvent system for whole cell biomass dissolution and NMR characterization. Employing this solvent system, semi-quantitative C-13-H-1 heteronuclear single quantum correlation (HSQC) spectroscopy of untreated, steam, dilute acid and lime pretreated poplar biomass samples was readily accomplished. In an effort to demonstrate the efficacy and usefulness of this fairly new characterization technique, relative spectral intensities of the untreated and pretreated biomass samples were evaluated and compared. From the relative signal intensities of hemicelluloses in each system it was observed that hemicelluloses are being removed in various pretreatment conditions, but complete dissolution of hemicellulose was observed only with acid pretreatment. The relative changes in lignin subunits after pretreatment were estimated from the volume integration of resolved cross peaks of various lignin subunits. The degradation of lignin was observed in all pretreatments, though more significant changes were noticed after dilute acid and lime pretreatment. HSQC analysis results were in agreement with the composition analysis of pretreated biomass samples. Thus, this methodology broadens the application of whole cell NMR analysis in biofuel research. (C) 2011 Elsevier Ltd. All rights reserved.