Abstract
An isocratic high performance liquid chromatographic (HPLC) method for the analysis of polymyxins B1 and B2 is described. The method uses a 25 cm Hypersil—ODS column, a mobile phase containing 22.5% acetonitrile (v/v) in an aqueous phase with tetramethylammonium chloride (TMAC), a flow rate of 1.09 ml/minute and a wavelength of 220 nm for detection. Complete resolution of B1 and B2, and their separation from all other components and/or impurities have been achieved in less than 23 minutes. The capability of the method for the determination of the polymyxin content in pharmaceutical preparations has also been demonstrated.