A dynamic ultrasonic extraction (DUE) coupled with on-line detection by spectrophotometry is proposed for the determination of total flavonoids (calculated against scutellarin) in Scutellaria barbata D. Don. The extraction was performed in a common self-made extraction vessel put in an ultrasonic bath and a peristaltic pump was used to deliver the extraction solvent. Several experimental parameters of DUE, including flow rate and concentration of extraction solvent, ultrasonic power and sample amount, were optimized. In this work an on-line method was provided with the advantages of minimum sample amount (5 mg), on-line monitoring the extraction process and time saving (12.5 min). Compared with off-line method, the proposed method would be more convenient for obtaining continuous measurements and rapid optimization of the extraction process.