Displacement chromatography of isomers and therapeutic compounds

Displacement chromatography was successfully used to separate a binary isomer mixture, epirubicin and doxorubicin, on Kromasil KR100-10 C-18 250X4.6 mm I.D. (10 mum) column. Displacement parameters such as the types and the concentrations of displacer, the composition and the flow rate of the mobile phase were critically examined in this study. The displacer employed was 30 mg/ml benzethonium chloride. Loading of feed at lower initial organic level of mobile phase coupled with displacement at higher organic level was found to give efficient separation. A 30-mg amount of binary isomer mixture was separated on an analytical column. The purification of epirubicin from the closely related impurities present in raw product solution by displacement chromatography was also investigated. The purity of epirubicin required was greater than 99% with a recovery of 60%. The results have indicated that this process made good use of the high feed load, low solvent costs, and high resolution characteristics of displacement chromatography and offered the chromatographic engineer a powerful tool for the preparative purification of therapeutic compounds. (C) 2002 Elsevier Science B.V. All rights reserved.