High-Pressure Liquid Chromatography Separation of Potential Impurities of Altretamine

Abstract

Two isocratic liquid chromatographic separation methods and UV detection were developed to allow for sensitive and specific analysis of potential impurities in Altretamine using a reversed phase C₁₈ column and mixtures of water-acetonitrile as mobile phase. Linear calibration curves for each of the possible contaminants of Altretamine, in the range of 0.25–125 μg/ml, were also obtained. The detection limit for all of the compounds (except cyanuric acid) was less than 0.25 μg/ml. Several Altretamine lots were examined and their impurities identified. Hydrolysis of cyanuric chloride to cyanuric acid was studied and shown to follow 1^st order kinetics. Evidence for the formation of chlorohydroxytriazine intermediates during the hydrolysis of cyanuric chloride to cyanuric acid is given.