Comprehensive Off-Line, Two-Dimensional Liquid Chromatography. Application to the Separation of Peptide Digests

Nicola Marchetti,† Jacob N. Fairchild, and Georges Guiochon*
Department of Chemistry, The University of Tennessee, Knoxville, Tennessee 37996-1600, and Division of Chemical Sciences, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831
Anal. Chem., 2008, 80 (8), pp 2756–2767
DOI: 10.1021/ac7022662
Publication Date (Web): March 21, 2008
Copyright © 2008 American Chemical Society
†

 Present address:  Department of Chemistry, University of Ferrara, via L. Borsari 46, 44100 Ferrara, Italy.

,
*

 To whom correspondence should be addressed. Tel.:  +1-865-974-0733. Fax:, +1-865-974-2667. E-mail:  guiochon@utk.edu.

Abstract

The separation of the peptide digests of myoglobin and bovine serum albumin was performed with an off-line combination of two commercial, conventional HPLC columns. The first column was packed with a strong ion exchanger and eluted with a KCl gradient. The second column was packed with particles of C18-bonded silica and eluted with an acetonitrile gradient. The conditional peak capacities of the 2D separations achieved exceed 7000 under the experimental conditions investigated. This performance is achieved at the cost of an analysis time of the order of 28 hours. Possible improvements to the separation method described here are discussed.

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History

  • Published In Issue April 15, 2008
  • Received for review November 2, 2007. Accepted January 25, 2008.

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