+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Mixed mode hydrophilic and ionic interaction chromatography rivals reversed phase liquid chromatography for the separation of peptides



Mixed mode hydrophilic and ionic interaction chromatography rivals reversed phase liquid chromatography for the separation of peptides



Journal of Chromatography 594(1-2): 75-86



Peptide separations based upon mixed-mode hydrophilic and ionic interactions with a strong cation-exchange column have been investigated. The peptide separations were generally achieved by utilizing a linear increasing salt (sodium perchlorate) gradient in the presence of acetonitrile (20-90%, v/v) at pH 7. The presence of acetonitrile in the mobile phase promotes hydrophilic interactions with the hydrophilic stationary phase, these hydrophilic interactions becoming increasingly important to the separation process as the acetonitrile concentration is increased. At acetonitrile concentrations of 20-50% (v/v) in the mobile phase, the peptides utilized in this study were eluted in order of increasing net positive charge, indicating that ionic interactions were dominating the separation process. Peptides with the same net positive charge were also well resolved by an hydrophilic interaction mechanism, being eluted in order of increasing hydrophilicity (decreasing hydrophobicity). At higher acetonitrile concentrations (70-90%, v/v), column selectivity was changed dramatically, with hydrophilic interactions now dominating the separation process. Under these conditions, specific peptides may be eluted earlier or later than less highly charged peptides, depending upon their hydrophilic/hydrophobic character. This mixed-mode methodology was compared to reversed-phase liquid chromatography of the peptides at pH 2 and pH 7. The results of this comparison suggested that mixed-mode hydrophilic-ion-exchange chromatography on a strong cation-exchange column rivals reversed-phase liquid chromatography for peptide separations.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 007556554

Download citation: RISBibTeXText

DOI: 10.1016/0021-9673(92)80314-k


Related references

Mixed mode hydrophilic and ionic interaction chromatography rivals reversed phase chromatography for the separation of peptides. Smith, J A And J E Rivier (Ed ) Peptides: Chemistry And Biology; Twelfth American Peptide Symposium, Cambridge, Massachusetts, Usa, June 16-21, 1991 Lviii+989p Escom Science Publishers B V : Leiden, Netherlands Illus 546-548, 1992

Comparison of reversed-phase liquid chromatography and hydrophilic interaction/cation-exchange chromatography for the separation of amphipathic alpha-helical peptides with L- and D-amino acid substitutions in the hydrophilic face. Journal of Chromatography. a 1009(1-2): 61-71, 2003

Preparation, characterization and application of a reversed phase liquid chromatography/hydrophilic interaction chromatography mixed-mode C18-DTT stationary phase. Talanta 146: 442-451, 2016

On-line comprehensive two dimensional separations of charged compounds using reversed-phase high performance liquid chromatography and hydrophilic interaction chromatography. Part II: application to the separation of peptides. Journal of Chromatography. a 1306: 27-36, 2014

Study of Separation and Identification of the Active Ingredients in Gardenia jasminoides Ellis Based on a Two-Dimensional Liquid Chromatography by Coupling Reversed Phase Liquid Chromatography and Hydrophilic Interaction Liquid Chromatography. Journal of Chromatographic Science 55(1): 75-81, 2016

New poly(ionic liquid)-grafted silica multi-mode stationary phase for anion-exchange/reversed-phase/hydrophilic interaction liquid chromatography. Analyst 137(11): 2553-2555, 2012

Preparation of a Monolithic Column with Mixed-Mode Stationary Phase of Reversed-Phase/Hydrophilic-Interaction for Capillary Liquid Chromatography. Journal of Separation Science 2018, 2018

Separation properties of novel and commercial polar stationary phases in hydrophilic interaction and reversed-phase liquid chromatography mode. Journal of Separation Science 31(9): 1492-1503, 2008

Separation and purification of the components in Trachelospermum jasminoides by two dimensional hydrophilic interaction liquid chromatography- reversed-phase liquid chromatography. Se Pu 35(6): 650-655, 2018

Comprehensive analysis of pharmaceutical products using simultaneous mixed-mode (ion-exchange/reversed-phase) and hydrophilic interaction liquid chromatography. Journal of Separation Science 37(16): 2138-2144, 2015

Separation of polar mushroom toxins by mixed-mode hydrophilic and ionic interaction liquid chromatography-electrospray ionization-mass spectrometry. Journal of Chromatographic Science 45(2): 104-111, 2007

Target separation of flavonoids from Saxifraga tangutica using two-dimensional hydrophilic interaction chromatography/reversed-phase liquid chromatography. Journal of Separation Science 2018, 2018

Orthogonal separation on one beta-cyclodextrin column by switching reversed-phase liquid chromatography and hydrophilic interaction chromatography. Talanta 81(4-5): 1870-1876, 2010

Chromatographic evaluation of a newly designed peptide-silica stationary phase in reverse phase liquid chromatography and hydrophilic interaction liquid chromatography: mixed mode behavior. Journal of Chromatography. a 1266: 43-52, 2013

Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension. Journal of Chromatography. a 1228: 242-249, 2012