A method for determination of palmatine and berberine from coptis chinensis franch by high performance liquid chromatography(HPLC) was developed. Optimun operating conditions were as follows: ODS C18 column. (4.6 250 mm.) CH3CN:0.05mol/LKH2PO4H2O(pH=2.5)=30:70(V/V) mobile phase with a flow rate of 0.6 mL/min. and using UV detector at 268 nm. The results of palmatine and berberine show that the calibration equition was C=1.696×10-5A+0.8401, R2=0.9999 and C=1.580×10-5A+0.5114, R2=0.9999. The reproducibility was good with RSD=0.015% and 0.14% respectivelv. The recovery was 99.09%～100.30% and 98.96%～101.86% respectively. The method is accurate, rapid and simple.