Flavonoids glycosides from ginkgo are widely applied in pharmaceutical, food, daily necessities and other areas due to their variety of biological activities. To overcome the shortcoming of current methods of time wasting, complex operation and expensive costs, an accuracy, low costs and fast determination method was established for flavonoids glycosides from ginkgo. According to the principle that flavonoids can form fluorescent chelate with Al3+, the detection condition was explored with rutin as standard. The results showed that the fluorimetric intensy of chelate of rutin with Al3+ would be stabilized and achieve maximum with λex=400 nm and λem=520 nm, in Al(NO3)3-(HAc-NaAc) reaction system for 1 500 s with pH 3.6. The linear regression equation y=29.92x+36.49 (R2=0.986) was deduced with the concentration of rutin and fluorescence intensy, and the linear range 1.8×10－6～3.2×10－5 mol·L-1. Flavonoids glycosides of cell suspension cultures from ginkgo biloba was detected by this method. The recovery experiments were also carried out with the average recovery rate of 101.3%. The advantages of high sensitivity, reproducibility, simple operation, and low costs were showed, indicating its good prospects.