File(s) under permanent embargo
Improving peak shapes with counter gradients in two-dimensional high performance liquid chromatography
journal contribution
posted on 2014-04-01, 00:00 authored by Paul Stevenson, Danielle Bassanese, Xavier ConlanXavier Conlan, Neil BarnettNeil BarnettTo achieve the greatest peak capacity in two-dimensional high performance liquid chromatography (2D-HPLC) a gradient should be operated in both separation dimensions. However, it is known that when an injection solvent that is stronger than the initial mobile phase composition is deleterious to peak performance, thus causing problems when cutting a portion from one gradient into another. This was overcome when coupling hydrophilic interaction with reversed phase chromatography by introducing a counter gradient that changed the solvent strength of the second dimension injection. It was found that an injection solvent composition of 20% acetonitrile in water gave acceptable results in one-dimensional simulations with an initial composition of 5% acetonitrile. When this was transferred to a 2D-HPLC separation of standards it was found that a marked improvement in peak shape was gained for the moderately retained analytes (phenol and dimethyl phthalate), some improvement for the weakly retained caffeine and very little change for the strongly retained n-propylbenzene and anthracene which already displayed good chromatographic profiles. This effect was transferred when applied to a 2D-HPLC separation of a coffee extract where the indecipherable retention profile was transformed to a successful application multidimensional chromatography with peaks occupying 71% of the separation space according to the geometric approach to factor analysis.
History
Journal
Journal of chromatography AVolume
1337Pagination
147 - 154Publisher
ElsevierLocation
Amsterdam, The NetherlandsPublisher DOI
ISSN
0021-9673Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2014, ElsevierUsage metrics
Categories
No categories selectedKeywords
two-dimensional high performance liquid chromatographynatural product separationscolumn efficiencysolvent strengthcounter gradientScience & TechnologyLife Sciences & BiomedicinePhysical SciencesBiochemical Research MethodsChemistry, AnalyticalBiochemistry & Molecular BiologyChemistryTwo-dimensional high performance liquidchromatographyBAND-BROADENING CONTRIBUTIONSINJECTION VOLUMEMOBILE-PHASESEPARATIONORTHOGONALITYCAPACITY
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC