Wydawnictwo Uniwersytetu Przyrodniczego w Poznaniu

Biblioteka Uniwersytetu Przyrodniczego w Poznaniu

Nauka Przyroda Technologie

Thomson Routers Master List

Scopus

Ebsco

Directory of Open Acces Journals

CBR- Open Access Journals in Poland

FOOD SCIENCE CENTRAL

Index Copernicus Journal Master List

Pubmed Medline

Polska Bibliografia Naukowa

Creaive Commons

Creative Commons licence CC BY-NC (Attribution-NonCommercial)
Issue 10 (2) 2011 pp. 213-221

Jerzy Dziuba, Piotr Minkiewicz, Damir Mogut

University of Warmia and Mazury in Olsztyn

Determination of theoretical retention times for peptides analyzed by reversed- -phase high-performance liquid chromatography

Abstract

 

Background. Peptides are important components of foods mainly due to their biological activity. The basic method of their identification is reversed phase high-performance liquid chromatography coupled with electrospray-ionization mass spectrometry (RP-HPLC-ESI-MS). Retention time (tR) prediction in silico is very helpful in analysis of multicomponent peptide mixtures. One of problems associated with RP-HPLC-ESI-MS is deterioration of mass spectra quality by trifluoroacetic acid (TFA). This problem can be avoided through the use of chromatographic columns designed for work with low TFA concentrations in mobile phase. The objective of this study was to determine the correlations between peptide retention times predicted with the use of a program available on-line and experimental retention times obtained using the column working with low TFA concentrations.
Material and methods. The set of synthetic peptides and bovine α-lactalbumin fragments (18 peptides) was used in the experiment. Theoretical retention times were calculated using Sequence Specific Retention Calculator (SSRC) program. The experimental retention times were measured via RP-HPLC-ESI-MS method using column working with low TFA content. The dependence between theoretical and experimental tR was expressed via empirical equations.
Results. The best correlation between theoretical and experimental retention times of peptides containing at least four amino acid residues has been obtained when third order polynomial (R² = 0.9536). Prediction quality for di- and tripeptides was significantly lower. The method described can be applied for cysteine-containing peptides although our sample preparation procedure did not include modification of this amino acid, taken into attention by SSRC program.
Conclusions. The results of this study validate the usefulness of a third degree polynomial as a simple function describing the correlation between peptide retention times predicted by an on-line application and experimental retention times. The above function can effectively predict retention times in situations when experimental conditions differ from the computational environment (various columns, mobile phase composition, use or resignation from chemical modifications during sample preparation, various HPLC equipments). On-line available tR predicting application with correction based on user’s own data may be a useful tool in food peptidomics.

 

Keywords: peptides, RP-HPLC, prediction of retention times, high-performance liquid chromatography, polynomial functions
pub/.pdf Full text available in english in Adobe Acrobat format:
http://www.food.actapol.net/issue2/volume/7_2_2011.pdf

For citation:

MLA Dziuba, Jerzy, et al. "Determination of theoretical retention times for peptides analyzed by reversed- -phase high-performance liquid chromatography." Acta Sci.Pol. Technol. Aliment. 10.2 (2011): 213-221.
APA Dziuba J., Minkiewicz P., Mogut D. (2011). Determination of theoretical retention times for peptides analyzed by reversed- -phase high-performance liquid chromatography. Acta Sci.Pol. Technol. Aliment. 10 (2), 213-221
ISO 690 DZIUBA, Jerzy, MINKIEWICZ, Piotr, MOGUT, Damir. Determination of theoretical retention times for peptides analyzed by reversed- -phase high-performance liquid chromatography. Acta Sci.Pol. Technol. Aliment., 2011, 10.2: 213-221.