OBJETCTIVE： To establish a method for the content determination of ethacridine lactate in Compound ethacridine ointment. METHODS： HPLC was performed on the column of Agilent ZORBAX SB-C18 with mobile phase of 0.1％ Octanesulfonic acid sodium solution-acetonitrile （70 ∶ 30，V/V） at a flow rate of 1.0 ml/min， the detection wavelength was 270 nm， the column temperature was 30 ℃， and the injection volume was 10 μl. RESULTS： The linear range of ethacridine lactate was 10.002-50.010 μg/ml（r＝0.999 9）； RSDs of precision， stability and reproducibility tests were less than 1％； recovery was 98.96％-100.36％ （RSD＝0.49％，n＝9）. CONCLUSIONS： The method is simple， accurate and reproducible， and can be used for content determination of ethacridine lactate in Compound ethacridine ointment.