Skip to main content
Article
Simultaneous separation of hydrophobic and hydrophilic peptides with a silica hydride stationary phase using aqueous normal phase conditions
Journal of Chromatography A (2011)
  • R. I. Boysen, Monash University
  • Y. Yang, Monash University
  • J. Chowdhury, Monash University
  • Maria T. Matyska-Pesek, San Jose State University
  • J. J. Pesek, San Jose State University
  • M. T.W. Hearn, Monash University
Abstract

The application of a silica hydride modified stationary phase with low organic loading has been investigated as a new type of chromatographic material suitable for the separation and analysis of peptides with electrospray ionization mass spectrometric detection. Retention maps were established to delineate the chromatographic characteristics of a series of peptides with physical properties ranging from strongly hydrophobic to very hydrophilic and encompassing a broad range of pI values (pI 5.5–9.4). The effects of low concentrations of two additives (formic acid and acetic acid) in the mobile phase were also investigated with respect to their contribution to separation selectivity and retention under comparable conditions. Significantly, strong retention of both the hydrophobic and the hydrophilic peptides was observed when high-organic low-aqueous mobile phases were employed, thus providing a new avenue to achieve high resolution peptide separations. For example, simultaneous separation of hydrophobic and hydrophilic peptides was achieved under aqueous normal phase (ANP) chromatographic conditions with linear gradient elution procedures in a single run, whilst further gradient optimization enabled improved peak efficiencies of the more strongly retained hydrophobic and hydrophilic peptides.

Disciplines
Publication Date
2011
Publisher Statement
SJSU users: use the following link to login and access the article via SJSU databases
Citation Information
R. I. Boysen, Y. Yang, J. Chowdhury, Maria T. Matyska-Pesek, et al.. "Simultaneous separation of hydrophobic and hydrophilic peptides with a silica hydride stationary phase using aqueous normal phase conditions" Journal of Chromatography A Vol. 1218 Iss. 44 (2011)
Available at: http://works.bepress.com/maria_matyska-pesek/11/