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Journal article
Aqueous size-exclusion chromatography of polyelectrolytes on reversed-phase and hydrophilic interaction chromatography columns
Journal of Chromatography A, v 1532, pp 161-174
12 Jan 2018
PMCID: PMC6605059
PMID: 29248345
Abstract
•Reversed-phase and HILIC columns are suitable for aqueous SEC.•The best performance is seen with medium polarity and polar stationary phases.•An electrolyte and organic modifier in the mobile phase minimize non-SEC effects.•Higher column temperatures reduce non-SEC interactions.•Low molar mass polyelectrolyte oligomers are more susceptible to non-SEC interactions.
The size-exclusion separation of a water-soluble polyelectrolyte polymer, sodium polystyrene sulfonate (NaPSS), was demonstrated on common reversed-phase (C18, C4, phenyl, and cyano) and hydrophilic interaction chromatography (HILIC) columns. The effect of common solvents – acetonitrile (ACN), tetrahydrofuran (THF), and methanol (MeOH), used as mobile phase modifiers – on the elution of NaPSS and the effect of column temperature (within a relatively narrow range corresponding to typical chromatographic conditions, i.e., 10 °C–60 °C) on the partition coefficient, KSEC, were also investigated. Non-size-exclusion chromatography (non-SEC) effects can be minimized by the addition of an electrolyte and an organic modifier to the mobile phase, and by increasing the column temperature (e.g., to 50 °C or 60 °C). Strong solvents such as THF and ACN are more successful in the reduction of such effects than is the weaker solvent MeOH. The best performance is seen on medium polarity and polar stationary phases, such as cyanopropyl- and diol-modified silica (HILIC), where the elution of the NaPSS polyelectrolyte is by a near-ideal SEC mechanism. Hydrophobic stationary phases, such as C18, C4, and phenyl, require a higher concentration of a strong solvent modifier (THF) in the mobile phase to reduce non-SEC interactions of the solute with the stationary phase.
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Details
- Title
- Aqueous size-exclusion chromatography of polyelectrolytes on reversed-phase and hydrophilic interaction chromatography columns
- Creators
- Anna M. Caltabiano - GlaxoSmithKlineJoe P. Foley - Drexel UniversityAndré M. Striegel - National Institute of Standards and Technology
- Publication Details
- Journal of Chromatography A, v 1532, pp 161-174
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemistry
- Web of Science ID
- WOS:000423894500018
- Scopus ID
- 2-s2.0-85038416532
- Other Identifier
- 991019168506804721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Chemistry, Analytical