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

Erik L Regalado; Joseph A Schariter; Christopher J Welch

doi:10.1016/j.chroma.2014.08.025

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

J Chromatogr A. 2014 Oct 10:1363:200-6. doi: 10.1016/j.chroma.2014.08.025. Epub 2014 Aug 14.

Authors

Erik L Regalado¹, Joseph A Schariter², Christopher J Welch³

Affiliations

¹ Merck Research Laboratories, Rahway, NJ 07065, USA. Electronic address: erik.regalado@merck.com.
² Merck Research Laboratories, Rahway, NJ 07065, USA.
³ Merck Research Laboratories, Rahway, NJ 07065, USA. Electronic address: christopher_welch@merck.com.

PMID: 25160950
DOI: 10.1016/j.chroma.2014.08.025

Abstract

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.

Keywords: Chiral separations; Closely related species; Complex mixtures; Method development; Two-dimensional HPLC; Warfarin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, High Pressure Liquid / methods*
Isomerism
Warfarin / analysis*
Warfarin / chemistry

Substances

Warfarin