Recebido em 23/7/08; aceito em 25/2/09; publicado na web em 3/7/09 VALIDATION OF METHODOLOGY FOR THE CHEMICAL CHARACTERIZATION OF SUGAR CANE BAGASSE. In this work, a methodology for the characterization of sugar cane bagasse was validated. Bagasse pre-treated with steam in a 5000 L reactor at a pressure of 15.3 kgf/cm 2 , during 7 min, was used to test the methodology. The methodology consisted of the hydrolysis of the material with H 2 SO 4 at 72% v/v, for the quantification of carbohydrates, organic acid, furfural and hydroxymethylfurfural by HPLC; insoluble lignin and ash by gravimetry; and soluble lignin by spectrophotometry. Linearity, repeatability, reproducibility and accuracy of the results obtained in two Research Laboratories were determined, and were considered to be suitable for the validation of the methodology.

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The influence of time (8-24 min), temperature (144-186 °C) and phosphoric acid concentration (0.05-0.20%, w/v) on the pretreatment of sugarcane bagasse in a 20 L batch rotary reactor was investigated. The efficiency of the pretreatment was verified by chemical characterization of the solid fraction of the pretreated bagasse and the conversion of cellulose to glucose by enzymatic hydrolysis. Models representing the percentage of cellulose, hemicelluloses, lignin, solubilized hemicellulose and the enzymatic conversion of cellulose to glucose were predictive and significant. Phosphoric acid concentration of 0.20% at temperature of 186 °C, during 8 and 24 min, was shown to be very effective in solubilizing hemicellulose from sugarcane bagasse, reaching solubilization of 96% and 98%, respectively. Relatively low amounts of inhibitors were produced, and the phosphoric acid remaining in the hemicellulosic hydrolysate is at adequate levels for supplying phosphorous requirement during subsequent fermentation.