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| Naji, M. |
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| Motta, Antonella |
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| Aletan, Dirar |
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| Mohamed, Tarek |
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| Ertürk, Emre |
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| Taccardi, Nicola |
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| Kononenko, Denys |
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| Petrov, R. H. | Madrid |
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| Alshaaer, Mazen | Brussels |
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| Bih, L. |
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| Casati, R. |
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| Muller, Hermance |
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| Kočí, Jan | Prague |
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| Šuljagić, Marija |
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| Kalteremidou, Kalliopi-Artemi | Brussels |
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| Azam, Siraj |
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| Ospanova, Alyiya |
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| Blanpain, Bart |
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| Ali, M. A. |
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| Popa, V. |
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| Rančić, M. |
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| Ollier, Nadège |
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| Azevedo, Nuno Monteiro |
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| Landes, Michael |
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| Rignanese, Gian-Marco |
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Broeckhoven, Ken
Vrije Universiteit Brussel
in Cooperation with on an Cooperation-Score of 37%
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Publications (4/4 displayed)
- 2024Detailed analysis of the effective and intra-particle diffusion coefficient of proteins at elevated pressure in columns packed with wide-pore core-shell particlescitations
- 2022Measurement of the molecular diffusion coefficient and the effective longitudinal diffusion under supercritical fluid chromatography conditions in packed bed columnscitations
- 2022Review of recent insights in the measurement and modelling of the B-term dispersion and related mass transfer properties in liquid chromatographycitations
- 2008Errors involved in the Existing B-term Expressions for the Longitudinal Diffusion in Fully Porous Chromatographic Media. Part II: Experimental data in Packed Columns and Surface Diffusion Measurements
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article
Detailed analysis of the effective and intra-particle diffusion coefficient of proteins at elevated pressure in columns packed with wide-pore core-shell particles
Abstract
<p>To determine the efficiency that can be obtained in a packed-bed liquid-chromatography column for a particular analyte, a correct determination of the molecular and effective diffusion coefficients (D<sub>m</sub> and D<sub>eff</sub>) of the analyte is required. The latter is usually obtained via peak parking experiments wherein the flow is stopped. As a result, the column pressure rapidly dissipates and the measurement is essentially conducted at ambient pressure. This is problematic for analytes whose retention depends on pressure, such as proteins and potentially other large (dipolar) molecules. In that case, a conventional peak parking experiment is expected to lead to large errors in D<sub>eff</sub>. To obtain a better estimate ofD<sub>eff</sub>, the present study reports on the use of a set-up enabling peak parking measurements under pressurized conditions. This approach allowed us to report, for the first time, D<sub>eff</sub> for proteins at elevated pressure under retained conditions. First, D<sub>eff</sub> was determined at a (average) pressure of about 105 bar for a set of proteins with varying size, namely: bradykinin, insulin, lysozyme, β-lactoglobulin, and carbonic anhydrase in a column packed with 400 Å core-shell particles. The obtained data were then compared to those of several small analytes: acetophenone, propiophenone, benzophenone, valerophenone, and hexanophenone. A clear trend between D<sub>eff</sub> and analyte size was observed. The set-up was then used to determine D<sub>eff</sub> of bradykinin and lysozyme at variable (average) pressures ranging from 28 bar to 430 bar. These experiments showed a decrease in intra-particle and surface diffusion with pressure, which was larger for lysozyme than bradykinin. The data show that pressurized peak parking experiments are vital to correctly determine D<sub>eff</sub> when the analyte retention varies significantly with pressure.</p>