The increasing interest in lignocellulose-based biorefineries boosts the further development of the needed pretreatment methods for preprocessing biomass. There are a large number of different processes that are being investigated; however research is made mostly based on different types of biomass with the same pretreatment or several modifications of the same process for a given type of biomass. In this work a comparison of promising chemical pretreatments using the same biomass was performed. Organosolv (OS), Steam (SE) and Liquid-Hot-Water (LHW) processes were used for the pretreatment of rye straw and the treated solids further enzymatically hydrolyzed. Best results for carbohydrate and lignin yield were found for the OS pretreatment followed close by the LHW and SE with similar results. All of the processes showed satisfactory performance for the pretreatment of lignocellulosic biomass for application in the second generation biorefinery.