title: Some factors that can lead to poor peak shape in hydrophilic interaction chromatography, and possibilities for their remediation.
abstract: Some factors which present difficulties for obtaining good peak shape in hydrophilic interaction chromatography (HILIC) were studied. The effect of injection solvent composition and volume was systematically investigated using a selection of weak and stronger basic compounds on a hybrid bare silica phase. Increasing the mismatch between the injection solvent (range 95-0% ACNv/v) and the mobile phase (maintained at 95% ACNv/v) gave increasing deterioration in peak shape. With the 2.1mm ID columns used, injections in the mobile phase of increasing volume (1-20 μL) gave poorer peak shape, but the magnitude of the effect was considerably smaller than that of solvent mismatch over this range. Some solute structural features such as galloyl (trihydroxy benzene), catechol (benzene diol) and phosphate (in nucleotides) gave serious peak tailing, attributed to interactions with metals in the stationary phase or the chromatographic hardware. These undesirable effects can be moderated by including complexing agents in the mobile phase, by changing the stationary phase chemistry, or by altering the mobile phase pH.
authors.affiliation: Centre for Research in Biosciences, University of the West of England, Frenchay, Bristol BS16 1QY, UK.
authors.forename: James C
authors.initials: JC
authors.lastname: Heaton
authors.affiliation: Centre for Research in Biosciences, University of the West of England, Frenchay, Bristol BS16 1QY, UK. Electronic address: david.mccalley@uwe.ac.uk.
authors.forename: David V
authors.initials: DV
authors.lastname: McCalley
mesh_terms: D002395:Catecholamines / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D062385:Hydroxybenzoates / Q000032:analysis; D009711:Nucleotides / Q000032:analysis; D012822:Silicon Dioxide / Q000737:chemistry; D012996:Solutions; D012997:Solvents
chemical_list: D002395:Catecholamines; D062385:Hydroxybenzoates; D009711:Nucleotides; D012996:Solutions; D012997:Solvents; D012822:Silicon Dioxide
keywords: Acids; Catecholamines; HILIC; Nucleotides; Peak shape