AIM: To establish HPLC method for separation and determination of rhein, emodin and crysophanol in Jiekangshu lotions(Radix et Rhizoma Rhei, Cortex Phellodendri, Radix Sophorae Flavescentis,etc.). METHODS: The chromatographic column: Symmetry C 18 column(3.9mm?150mm,5?m) and in advance column: Symmetry C 18 (5?m) were used with the mobile phase of methanol-water-phosphate acid(80∶20∶0.1), 1.0mL?min -1, flow rate, column temperature at 24℃ and detection wavelength at 432nm. RESULTS: Rhein, emodin and chrysophanol in Jiekangshu Lotion showed the good linear relationships at the range of 7.60～ 38.00?g?mL -1 (r= 0.9994), 1.75～8.75?g?mL -1 (r=0.9999), 2.40～12.02?g?mL -1 (r=0.9999), respectively. Their average recoveries: Rhein was 98.02%, Emodin 96.63% and Chrysophanol 97.05%, respectively. CONCLUSION: This method is simple. The results are accurate and reproducible.It can be used in the quality control of Jiekangshu lotion.