AIM: To establish the method for determination of gentiopicrin, palmatine hydrochloride and berberine hydrochloride in Ertong Ⅱ Oral Liquid (Cortex Phellodendri, Radix Gentianae, Rhizoma Anemarrhenae, etc.) by RP HPLC. METHODS: The column was a Diamonsil TM C 18 column (250mm?4.6mm, 5?m); the mobile phase was CH 3CN H 3PO 4 C 6H 15 N H 2O; The ultraviolet detection was set at 270nm; the flow rate was 1mL?min -1 . RESULTS: The linear ranges of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 72.8~728?g?mL -1 ( r=0.9998 ), 7.55~75.5?g?mL -1 ( r=0.9998 ), 12.45~124.5?g?mL -1 ( r = 0.9999 ), respectively. The average recoveries ( n =6) of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 98.17%, 98.78%, 98.60%; RSD were 1.60%, 1.35%, 1.25%, respectively. CONCLUSION: The method is accurate, reproducible and highly selective, thus suitable for quality control of Er Tong Ⅱ Oral Liqual.