Abstract: In this study,determination methodology of total flavonoids from Lycium barbarum was optimized employing decolorization,extraction solvent,and extraction method. The optimal conditions were determined as follows: the Lycium barbarum was extracted with reflux by 95% ethyl alcohol.Subsequently,the obtained extracting solution was refrigerated to remove the precipitate and the up- phase was concentrated. Furthermore,the concentrated solution was purified by extracting with absolute methanol. The purified solution was subsequently used for detecting the total flavonoids.With the optimized determination methodology,the average content of total flavonoids was 1.134% deriving from Ningxia Lycium barbarum while 1.020% from Sinkiang,and the variation range were 0.851% to 1.466% and 0.888% to 1.241%,respectively.This research can effectively remove the water soluble pigment which influenced the determination accuracy of total flavonoids.