Abstract: The feasibility of using hyperspectral imaging at near infrared wavelength range( 900 ~ 1700nm) for determination of total flavonoid content distribution on ginkgo biloba leaves was investigated. 120 fresh ginkgo leaves were collected as research samples. After hyperspectral image acquisition and pre- processing,standard destructive method was used to determine the content of total flavonoid content.The average spectra obtained from the region of interest( ROI) in ginkgo leaf images were used for model development.Principal component analysis( PCA) was performed on the average spectrum,multi- linear regression( MLR) was used to build calibration models relating the spectra and total flavonoid content( r = 0.9219).The calibration model was used to predict the total flavonoid content of each pixel in the hyperspectral image.This enables construction of a distribution map of total flavonoid content on the ginkgo leave image. The results indicated that hyperspectral imaging could offer an effective method for flavonoid concentration analysis.