An enzymatic hydrolysis - continuous flow method for the determination of starch in tobacco was developed. The tobacco products were smashed and filtered by 100 mesh sieve. The water-soluble sugars in tobacco were extracted with an 85% aqueous ethanol distillate. The sample powder was heated in boiling water for gelation of starch, and then the strong base was added in solution to destroy the structure of the starch further. In the presence of enzyme amyloglucosidase (AGS), the starch in tobacco was hydrolyzed to glucose at pH 4.6 in water bath at 56 °C for 90 minutes. The final solvent after volume adjustment was analyzed through continuous flow analyzer to determine the content of the glucose. The content of the starch can be calculated with that of the glucose. The recovery of the starch was 96.6% and the relative standard deviations (n=7) were in the range of 1.52 ~ 4.08%, indicating that the method had the advantage of stability and exactness. The method was compared with the enzymatic method (ISO 8451:91), the conventional acidic hydrolysis and iodine colorimetric methods. The contents of the starch in 18 tobacco samples were also determined.