Abstract: Fructosazine content in tobacco with ultrasonic extraction-high performance liquid chromatography method was established.Extract processes were as follows:extraction agent for pure water, extraction time 40min, liguid solid ratio 50:3 (mL/g) .Chromatographic conditions were separation column Waters Carbohydrate Analysis (3.9mm×300mm) , mobile phase acetonitrile:water (V/V) 80:20, flow velocity 1.0mL/min, room column temperature, wavelength detection 275nm, sample volume 10μL.The results showed that the correlation coefficient of standard curve was 0.9969, the linear ranged of 6 ～ 14μg/mL, fructose in the average rate of recovery was 98.7%, the relative standard deviation was less than 3%, the detection limit was 0.5μg/mL in tobacco.Fructosazine in the tobacco was about 0.02% in the mass percent concentration.This method was suit for analyzing the content of fructosazine to tobacco samples in quantitative.