Article (Scientific journals)
Gradient-elution parameters in capillary liquid chromatography for high-speed separations of peptides and intact proteins
Vaast, A.; Tyteca, Eva; Desmet, G. et al.
2014In Journal of Chromatography. A, 1355, p. 149-157
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Keywords :
Gradient focusing; High-throughput analysis; Linear solvent-strength model; Neue-Kuss model; Proteomics; Fused silica; Liquid chromatography; Molecular biology; Phase separation; Solvents; Capillary liquid chromatography; High degree of accuracy; High-speed separation; Mobile-phase composition; Retention time prediction; Reversed-phase separation; Peptides; Animals; Chromatography, High Pressure Liquid; Nonlinear Dynamics; Proteins; Silicon Dioxide
Abstract :
[en] This contribution relates to the assessment of gradient-elution parameters in capillary liquid chromatography affecting the peak widths in the reversed-phase separation of peptides and intact proteins. Gradient separations were performed using both a poly(sytrene-co-divinylbenzene) monolithic column and a microparticulate fused-core column (silica C18, 2.7μm). The applicability of the conventional linear (LSS) and non-linear solvent-strength model (Neue-Kuss) were investigated to describe the retention behaviour of the compounds as a function of the mobile-phase composition. This was performed by using a wide range of gradient conditions, including different gradient slopes (β, ranging from 0.05 to 0.65min-1) and mobile-phase compositions (δφ, i.e. gradient span). Although the LSS-model provided accurate retention time predictions (<1.3% deviation) of scouting runs with more conventional gradient slopes, the prediction of high-speed separations with a high degree of accuracy (<2%) could only be obtained with the non-linear model. The solvent-strength parameters resulting from the use of both models, as well as the retention factors at the moment of elution (ke), further served as input parameters to assess the influence of the gradient slope on the expected peak-compression effects in gradient mode, with a focus on high-speed separations. The importance of the correct model choice was emphasized in terms of compression; while the LSS-model lead to the conclusion of peak broadening rather than peak sharpening, the use of a more accurate non-linear model showed the existence of peak compression effect. The results presented in this manuscript show the occurrence of gradient-related focusing effects, which appear to be more prevalent for extremely fast separations. © 2014 Elsevier B.V.
Disciplines :
Chemistry
Author, co-author :
Vaast, A.;  Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium
Tyteca, Eva ;  Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Analyse, qual. et risques - Labo. de Chimie analytique
Desmet, G.;  Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium
Schoenmakers, P. J.;  University of Amsterdam, Van't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, Science Park 904, 1098 XH Amsterdam, Netherlands
Eeltink, S.;  Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium
Title :
Gradient-elution parameters in capillary liquid chromatography for high-speed separations of peptides and intact proteins
Publication date :
2014
Journal title :
Journal of Chromatography. A
ISSN :
0021-9673
eISSN :
1873-3778
Publisher :
Elsevier
Volume :
1355
Pages :
149-157
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 30 March 2017

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