［Objective］The research aimed to establish the method of determining the total flavone content in Japanese hop leaf by fluorescence spectrophotometry. ［Method］The effective components were extracted by ultrasonic with 75% ethanol at 60 ℃ from J. hop leaf, with rutin as standard sample, 410 and 534 nm were selected as excitation and emission wavelengths to study the effects of solvent, pH value and standing time on fluorescence intensity. ［Result］ The best determining conditions were as follows: The suitable addition amount was 4 ml for 10%Al(NO3)3, the addition of acetic acid-sodium acetate buffer with pH value of 4.1 was 2.5 ml,60% ethanol was selected as solvent and the suitable standing time of solution was 25 min. The total flavone content detected by this method in wild J. hop leaf was up to 52.98 mg/g and its detection limit was 9.43×10-7 mol/L. The fluorescence intensity had a good linear relationship with rutin concn. in the range of 1.55×10-6 - 1.31×10-5 mol/L with the regression equation of y=2.816 x+0.291, correlation coefficient of 0.996 6, average recovery of 100.5% and RSD of 0.54%. ［Conclusion］The method of determining the total flavone content in J. hop leaf by fluorescence spectrophotometry had relatively high precision and accuracy and was easy to operate and low in cost._