Efficient characterization of lignocellulosic biomass is essential for its utilization in various applications. Conventional analysis involves separate processing of cellulose and hemicellulose through acid hydrolysis and acid methanolysis, respectively, followed by HPLC-RID or GC-MS analysis [1,2]. These methods require lengthy procedures, multiple sample preparation steps and complex derivatization for quantification. Lignin content is determined independently through Klason lignin analysis, with acid-insoluble lignin measured gravimetrically and acid-soluble lignin quantified via UV spectroscopy [1].This study presents an optimized approach that simplifies the process by using a single sample preparation step. Acid hydrolysis was performed to release sugars and lignin components simultaneously from sawdust. The solid residue was used for Klason lignin determination, while the hydrolysate was analyzed for acid-soluble lignin using UV spectroscopy. The same hydrolysate was then utilized for sugar quantification via quantitative ¹H NMR spectroscopy, eliminating the need for external calibration standards or complex derivatization [3]. Comparative analysis highlighted variations in sugar recovery and profiling across techniques, demonstrating the efficiency of NMR in biomass characterization. This streamlined method reduces analysis time, minimizes reagent consumption, and offers a cost-effective alternative to conventional approaches.