In the theory of Korean medicine, Scrophulariae Radix (SR) can clear away heat and cool the blood, nourish yin and promote the production of the body fluids, relieve toxin and benefit the throat. The present study was carried out to investigate effects of SR on allergic contact dermatitis (ACD) induced by 2,4-dinitrochlorobenzene (DNCB) in mice. In this experiment, effects of SR on clinical aspects on the skin, histopathological changes such as spongiosis, mast cell distribution, immune cell infiltration in tissue, spleen/body ratio and production levels of serum cytokines were investigated in vivo. In addition, effects on cell viability and release of β-hexosaminidase and histamine were also investigated in vitro. In results, SR treatment diminished erythema, desquamation and keratosis which were induced by repeated painting of DNCB. Spongiosis and edema were diminished by painting of SR in histopathological observation, infiltrations of mast cell and monocytes were also decreased in SR group. In addition, spleen/body ratio was lowered compared to ADC control group. Production level of IFN-γ in serum was decreased, but level of IL-4 did not affected by SR. Finally, more than 400 ㎍/ml of SR treatment groups showed decreased cell viabilities in RBL-2H3 cells. Treatment with over 200 ㎍/ml of SR decreased β-hexosaminidase release, and treatment with over 400 ㎍/ml decreased histamine release in vitro. In conclusion, these data suggest that SR can decrease symptoms of ACD, then SR is useful to treat patient with ACD.