Abstract: A flow analysis method for determining the cellulose in tobacco was developed. Cellulose was hydrolyzed by cellulase and the formed glucose was determined by flow analyzer. The results showed that:1) The optimal hydrolysis conditions were:tobacco powder 0.25 g, buffer solution 60 mL, enzyme 0.25 g, hydrolysis temperature 37℃, hydrolysis time 24 hours. 2)The relative standard deviation (RSD) of the method was 2.40% with the recoveries from 96.7% to 103.8% and the limit of detection of 0.11 mg/mL. This method is suitable for determining the content of cellulose in tobacco.