The aim of this study was to determine the suitability of surfactant to extract higher phenolic compound, flavonoid and antioxidant activity from Tartary buckwheat and evaluate the potentiality of surfactant as a screening agent for breeding purpose. Primarily, we employed two types of surfactant (Hydrophilic: Tween 20 and Lipophilic: Span 80) to select the suitable surfactant agent for the extraction of optimum bioactive compounds. Between two surfactants, Tween 20 showed highest efficiency at 4 mM concentration to extract total phenolic content (TP), total flavonoid (TF) and antioxidant activity (AA). Tween 20 at 4 mM concentration was fixed for further analysis along with hot water (90℃) treatment as a control. In our findings, highest TP (118 ㎎/g), TF (38 ㎎/g) and AA (76%) was achieved in KW21 and KW22 among the fifteen accessions of Tartary buckwheat. In other way, TP, TF and AA was 200%, 120% and 110% higher in surfactant formulation compared with control treatment, respectively.