Paclitaxel in southern yew tree was quantitatively determined by high performance liquid chromatography (HPLC) with ODS-C₁₈ column. A mixture of CH₃OH-H₂O-CH₃COOH(volume ratio: 55 : 44 : 1) is used as mobile phase and UV detection is carried out at 227 nm, and the column temperature is 20°C. The results show that there is a good linear relationship between the area of paclitaxel and the concentration of the sample in the range 50–500 mg/L for paclitaxel. The corresponding regression equation is Y=13021.7+1.01×10⁶X, r=0.9990. The average recovery is 95.3% and the relative standard deviation is 2.08%.