Ozonolysis, as a lignocellulosic biomass pretreatment, goes back to 80s; however, in the last years it is becoming widespread again owing to its efficiency and mild operation conditions. Ozone reacts preferably with lignin than carbohydrates, promoting biomass destructuration and delignification, and so the sugar release by enzymatic hydrolysis.The hydrolysate from pretreated biomass has being used as sugars source for secondgeneration fuels production, mainly ethanol, methane and hydrogen. Short-chain carboxylic acids are the main inhibitory compounds generated, being properly removed by water washing. The most common inhibitory compounds reported for other pretreatments, furfural and HMF (5-hydroxymethylfurfural) † , are not found in ozonepretreated hydrolysates. Composition of pretreated biomass and ozone consumption depends on several process parameters: reactor design, moisture content, particle size, pH, reaction time, ozone/air flow and ozone concentration. Additional studies are necessary to clarify process parameters effect and to optimize the process to achieve high yields with economic feasibility.