This research evaluates the impact of different delignification pretreatments (enzymatic and organosolv) on the crystallinity and enzymatic hydrolysis of harvested sugar cane residues. The study uses the Crystallinity Index (CrI), Relative Number of Intensity (Ir), degree of cellulose mercerization (IIC-%), and Global Index of Saccharification (GIS) as parameters to assess six different substrates derived from sugar cane residues (leaves and tops) treated by various processes. Spectroscopic techniques, including Fourier Transform infrared spectroscopy (FTIR), X-ray diffraction, and scanning electron microscopy (SEM), are employed for characterization. Substrates pretreated only with organosolv without further treatment show good enzymatic hydrolysis and high CrI, likely due to increased crystallinity by removing amorphous material. The results indicate that pretreatment methods significantly affect the crystalline structure and enzymatic hydrolysis, with organosolv pretreatment effectively removing lignin and hemicellulose, leading to increased crystallinity and improved hydrolysis efficiency.This research evaluates the impact of different delignification pretreatments (enzymatic and organosolv) on the crystallinity and enzymatic hydrolysis of harvested sugar cane residues. The study uses the Crystallinity Index (CrI), Relative Number of Intensity (Ir), degree of cellulose mercerization (IIC-%), and Global Index of Saccharification (GIS) as parameters to assess six different substrates derived from sugar cane residues (leaves and tops) treated by various processes. Spectroscopic techniques, including Fourier Transform infrared spectroscopy (FTIR), X-ray diffraction, and scanning electron microscopy (SEM), are employed for characterization. Substrates pretreated only with organosolv without further treatment show good enzymatic hydrolysis and high CrI, likely due to increased crystallinity by removing amorphous material. The results indicate that pretreatment methods significantly affect the crystalline structure and enzymatic hydrolysis, with organosolv pretreatment effectively removing lignin and hemicellulose, leading to increased crystallinity and improved hydrolysis efficiency.