A two-stage pretreatment approach, employing steam followed by organosolv treatment, was assessed for its ability to fractionate and recover most of the hemicellulose, lignin and cellulose components of poplar wood chips. A mild steaming stage was initially used to maximise hemicellulose sugar recovery, with 63% of the original xylan solubilised and recovered after this stage and close to 90% recovered in total. Rather than hindering subsequent organosolv delignification, the prior steam treatment enhanced lignin solubilisation with more than 66% of the original lignin removed after the two-stage pretreatment. The extracted lignin contained at least equal or greater amounts of functional groups as compared to the lignin solubilised after a single-stage organosolv pretreatment. More than 98% of the original cellulose was recovered after the two-stage pretreatment and 88% of the cellulose could be hydrolysed to glucose at enzyme loading of 5FPU/g cellulose after 72h.