Investigation of two-dimensional high performance liquid chromatography approaches for reversed phase resolution of warfarin and hydroxywarfarin isomers

Several offline and online 2D HPLC methods were investigated for the reversed phase resolution of a complex mixture of closely related warfarin and hydroxywarfarin isomers. By combining reversed phase achiral/chiral HPLC separation with UV-triggered fraction collection and subsequent chiral/achiral reversed phase HPLC analysis of collected fractions, complete resolution of all 12 components of the mixture was possible. In addition, a faster method was developed from online 2D HPLC analysis where multicomponent fractions from the first dimension are simultaneously chromatographed in the second dimension. (C) 2014 Elsevier B.V. All rights reserved.