Predictive Liquid Chromatography of Peptides Based on Hydrophilic Interactions for Mass Spectrometry-Based Proteomics
- Authors: Lobas A.A.1, Levitsky L.I.1, Fichtenbaum A.2, Surin A.K.3, Pridatchenko M.L.1, Mitulovic G.2, Gorshkov A.V.4, Gorshkov M.V.1
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Affiliations:
- Tal’roze Institute for Energy Problems of Chemical Physics
- Medical University of Vienna
- Institute of Protein Research
- Semenov Institute of Chemical Physics
- Issue: Vol 72, No 14 (2017)
- Pages: 1375-1382
- Section: Articles
- URL: https://journals.rcsi.science/1061-9348/article/view/182740
- DOI: https://doi.org/10.1134/S1061934817140076
- ID: 182740
Cite item
Abstract
High-performance liquid chromatography (HPLC) is widely used for separation of complex peptide mixtures before mass spectrometry-based proteome analysis. In this analysis, reversed phase HPLC (RPHPLC) using non-polar stationary phases such as surface-modified silica containing alkyl groups (e.g., C18) is typically employed. Because of the high heterogeneity of proteomic samples, multidimensional separation approaches gained increasing attention recently to tackle this complexity and extremely high range of concentrations. In two-dimensional liquid chromatography, hydrophilic interaction chromatography (HILIC) is often a method of choice for combination with RP-HPLC because it uses reversed-phase type eluents and allows efficient separation of polar peptides. Due to the high degree of orthogonality in this two-dimensional separation space, it is tempting to develop approaches for predicting peptide retention times for HILIC-based separations similar to the ones for RP-HPLC. Recent successful efforts in this area were focused on developing retention coefficient (RC)-based approaches. Herein, we explored the feasibility of using a statistical thermodynamic model for prediction of peptide retention times in HILIC separations and determined the phenomenological parameters of the model for a bare silica column. The performance of the developed model was tested using HPLC-MS analysis of a set of synthetic peptides, as well as a tryptic peptide mixture.
Keywords
About the authors
A. A. Lobas
Tal’roze Institute for Energy Problems of Chemical Physics
Email: mike.gorshkov@gmail.com
Russian Federation, Moscow, 119334
L. I. Levitsky
Tal’roze Institute for Energy Problems of Chemical Physics
Email: mike.gorshkov@gmail.com
Russian Federation, Moscow, 119334
A. Fichtenbaum
Medical University of Vienna
Email: mike.gorshkov@gmail.com
Austria, Vienna, 1090
A. K. Surin
Institute of Protein Research
Email: mike.gorshkov@gmail.com
Russian Federation, Pushchino, Moscow oblast, 142290
M. L. Pridatchenko
Tal’roze Institute for Energy Problems of Chemical Physics
Email: mike.gorshkov@gmail.com
Russian Federation, Moscow, 119334
G. Mitulovic
Medical University of Vienna
Email: mike.gorshkov@gmail.com
Austria, Vienna, 1090
A. V. Gorshkov
Semenov Institute of Chemical Physics
Email: mike.gorshkov@gmail.com
Russian Federation, Moscow, 119991
M. V. Gorshkov
Tal’roze Institute for Energy Problems of Chemical Physics
Author for correspondence.
Email: mike.gorshkov@gmail.com
Russian Federation, Moscow, 119334