Resolving Co-Elution Problems of Components in Complex Mixtures by Multiple Heart-Cutting 2D-LC

Multiple heart-cutting (MHC) 2D-LC was applied to resolve challenging peak co-elutions in a pesticidal natural product-derived extract, which exhibited a complex impurity profile. While the 1D separation using sub-2 μm particle column technology provided good separation efficiency, additional selectivity was required to separate overlapping peaks in several chromatographic regions. MHC 2D-LC using C18 columns in both dimensions, but different pH in the mobile phase, provided full separation of the components of interest. This article describes how the MHC 2D-LC technology has been applied for optimized analysis of target components as well as being a rapid tool for peak purity evaluation. It also discusses strategic benefits of the technology regarding method development for separation of very complex samples.