Objective:To develop a HPLC method for determination of Podophyllotoxin in two preparations.Methods:HPLC was carried out,using a Dikma diamonsil C18 column(4.6mm?150mm,5?m)with a column temperature at 25℃,the methanol-water as mobile phase(using gradient)with flowing rate 1.0 mL/min.The detection wavelength was at 292 nm.Results:The method had good linear relationship within the range of 11.8-590?g/mL for Podophyllotoxin,with the correlation coefficients 0.9999.The results of the average recovery of Youdi liniment and Podophyllotoxin tincture were 99.38%,99.62%.Conclusion:The rapid,accurate and reproducible RP-HPLC method was useful for quantification of Podophyllotoxin in Youdi Liniment and Podophyllotoxin Tincture.