Protamine sulphate, which has the property of neutralising heparin, is determined in pharmaceutical formulations using spectrophotometric (BP 1995) or biochemical methods (USP XXIII 1995). Accuracy of these methods is not very high. We applied the HPLC technique for the assay of protamine sulphate in a gel formulation. The assay was carried out on a diol-type column with a mobile phase containing 8% acetonitrile in 0.15% trifluoroacetic acid at pH 2.5. The flow rate was 1.0 ml min(-1). The results obtained show that HPLC can be used for the determination of protamine sulphate in pharmaceutical preparations. The method is rapid and more accurate than those described until now.
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