Atmospheric pressure O₂ plasma was used to produce ozone in order to treat sugarcane bagasse as a function of particle sizes. The fixed bagasse moisture content was 50%. The delignification efficiency had small improvement due to ozonation process as a function of particle size, varying from 75 up to 80%. Few amounts of hemicellulose were removed, but the ozonation has not been affected significantly with particle size variance as well (from 30 up to 35%). The cellulose presented some losses below 1.0 mm size (8-15%) which was an unexpected result. The conversion of cellulose content into free sugar has shown a significant increase as the particle size has diminished as well. The best condition of the bagasse particle size was for 0.08 mm. For this case, a great quantity of cellulose (78.8%) was converted into glucose. Optical absorption spectroscopy was applied to determine ozone concentrations in real time where the samples with typical bagasse particle sizes equal or below to 0.5 mm had shown a better absorption of ozone in comparison with greater particle size samples.