Objectives We performed the simultaneous analysis for three compounds in Cheongsimyeonja-tang (CSYJT) and evaluated its anti-obesity effect. Methods The column for separation of three compounds was used Gemini $C_{18}$ column and maintained at $40^{circ}C$. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 275 nm. The injection volume was $10{mu}L$. 3T3-L1 preadipocytes were differentiated into adipocytes by adding insulin, dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) for 8 days in the absence or presence of CSYJT. Anti-obesity effects of CSYJT were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results Calibration curves were acquired with $r^2$ >0.9999. The contents of baicalin, wogonoside and baicalein in CSYJT were 14.54-14.65 mg/g, 5.24-5.27 mg/g and 0.01-0.02 mg/g, respectively. CSYJT showed inhibitory effect on lipid accumulation and GPDH activity in the differentiated 3T3-L1 cells. Furthermore, CSYJT significantly decreased contents of triglyceride and leptin production in 3T3-L1 adipocytes. Conclusions These results will be helpful to improve quality control and anti-obesity effect of Taeeumin CSYJT.