High-performance liquid chromatography packing materials for the analysis of small molecules in biological matrices by direct injection

Abstract

The increasing demand on high-performance liquid chromatography to resolve mixtures of closely related components in complex biological matrices in less time with higher precision has led to the development of a variety of new high-performance liquid chromatography columns, which eliminate the need for sample preparation. These packings isolate small molecules from biological macromolecules on direct sample injection by exerting two separation mechanisms. They allow elution of all sample macromolecules with high recovery in one peak at the extraparticulate void, because of size-exclusion interactions with hydrophilic outer particulate surfaces. Simultaneously, these packings allow permeation and partitioning of small molecules on bonded-phases which are protected from contamination by macromolecules. The names given to these new packings include “internal surface reversed-phase”, “shielded hydrophobic phase”, “semipermeable surface”, “dual zone material” and “mixed-functional phases”. The fundamental principles behind each of the design concepts are reviewed, and applications are cited.