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Proceedings of the Estonian Academy of Sciences

ISSN 1736-7530 (electronic)   ISSN 1736-6046 (print)
Formerly: Proceedings of the Estonian Academy of Sciences, series Physics & Mathematics and  Chemistry
Published since 1952

Proceedings of the Estonian Academy of Sciences

ISSN 1736-7530 (electronic)   ISSN 1736-6046 (print)
Formerly: Proceedings of the Estonian Academy of Sciences, series Physics & Mathematics and  Chemistry
Published since 1952
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Behaviour of proteins on reversed-phase supports during high performance liquid chromatography on C18 stationary phase; pp. 36–42

(Full article in PDF format) doi: 10.3176/proc.2015.1.05


Authors

Oksana Rotkaja, Jelena Golushko, Peteris Mekss

Abstract

The separation of high-molecular compounds is very difficult, if possible at all, under isocratic conditions. For this gradient elution is needed. The theory of gradient elution for small molecules is well established; however, its applications to reversed-phase gradient separations of biopolymers are not straightforward because of specific problems, such as slow diffusion, limited accessibility of the stationary phase for larger molecules, or possible sample conformation changes during the elution.
We used high performance liquid chromatography to investigate the reversed-phase chromatographic behaviour of 14 proteins. The first step was the determination of the experimental data, and then these data were used to predict gradient retention times. A water–organic solvent–trifluoroacetic acid system was used to examine the influence of experimental parameters. The chromatographic results from four C18-chain-length supports were comparable.

Keywords

high-performance liquid chromatography, mobile phase composition, protein, reversed phase.

References

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Current Issue: Vol. 68, Issue 3, 2019




Publishing schedule:
No. 1: 20 March
No. 2: 20 June
No. 3: 20 September
No. 4: 20 December