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 °C, 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.