An isocratic Reversed - Phase High Performance Liquid Chromatographic (RP -HPLC) method is presented for the simultaneous determination of atropine sulphate and scopolamine-N-butylbromide. The analytes were separated on a Brownlee RP-18, Spheri-5, 220 × 4.6mm I.D. column with a mobile phase consisting of 0.05M ammonium acetate - methanol-acetonitrile (48:32:20 v/v). Flufenamic acid was used as internal standard and the analysis was completed within approximately 10 rnin. The detection was performed with a variable wavelength UV - visible detector at 220 nm resulting in detection limits of 18 ng of each alkaloid per 10 μl injection.