The effect of different treatments on total flavonoids in Ginkgo biloba.leaves was studied.Three different methods including damp-heat, oxidation and bio-fermentation were selected to treat with Ginkgo biloba leaves.The content of total flavonoids in Ginkgo biloba leaves was determined by spectrophotometer.The results showed that the content of total flavonoids in Ginkgo biloba leaves treated by bio-fermentation has reached 7.97%,which was higher than other methods.Therefore bio-fermentation method has significant impact on flavonoid and it could increase the total content of flavonoids in Ginkgo biloba leaves.