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. The peak area ratios of each analyte relative to internal standard showed a linear relationship with concentrations between 1.81 - 90.4 ng/μl for atropine sulphate and 1.87 - 104 ng/μl for scopolamine-N-butylbromide. The average relative standard deviation for injection reproducibility was approximately 2%. The method was applied to the direct determination of scopolamine-N-butybromide in tablets, suppositories and ampoules and of atropine sulphate in ophthalmic solution as well as to the simultaneous determination of both alkaloids in human serum and urine samples after a Solid - Phase Extraction (SPE) clean-up step.
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Papadoyannis et al. (1994) studied this question.
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