Selectivity is key to achieve assay specificity and accuracy in bioanalytical applications.
It can be introduced on various levels of an analytical process, in the course of sample
preparation, by derivatization, chromatographic separation, two-dimensional
separations, ion-mobility spectrometry and mass spectrometry. Chiral stationary
phases (CSPs) enrich the column toolbox in liquid chromatography. They are
established tools for enantiomer separations of chiral drugs, but have also other
applicabilities beyond the field of enantiomer separations. The majority of chiral
selector chemistries of the columns which we use nowadays have been developed in
the 1980s/1990s, comprising polysaccharides, proteins, chiral synthetic polymers,
cyclodextrins, macrocyclic glycopeptides, chiral crown ethers, synthetic low molecular
selectors, chiral ligand exchange and cinchona alkaloid derivatives (chiral ionexchangers).
Core-shell and sub-2μm particle CSPs paved the way to UHPLC chiral
separations and their use in achiral UHPLC separations. In this presentation, the
potential of chiral stationary phases in metabolomics and lipidomics, for therapeutic
peptide analysis, and nucleic acid pharmaceuticals will be discussed. When it comes to
real bioanalytical applications, chromatographic selectivity is usually combined with
mass spectrometry (MS) to cope with assay specificity limitations in presence of real
matrix and insufficient chemoselectivity. Smart MS designs can work in concert with
CSPs for challenging separations such as in the analysis of
phosphatidylinositolphosphate constitutional isomers which are regulators of cell
membrane dynamics. Multidimensional LC combining achiral chemoselectivity with
stereoselectivity of CSPs is another option to deal with complex sample mixtures. Its
potential will be illustrated for absolute configuration determination of a natural product
with tetrapeptide structure. Chiral columns have some limitations regarding their
chemoselectivity. It often happens that enantiomers are well resolved but structural
analogs or constitutional isomers overlap. It will be discussed how ion mobility mass
spectrometry (IM-MS) coupling can combine enantioselectivity of CSPs with
diastereomer/constitutional isomer selectivity of IMS for establishing enantioselective
amino acid profiles in about 1 min.
