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Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides

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Hydrodynamic chromatography of macromolecules using polymer monolithic columns.

Rob Edam¹, Sebastiaan Eeltink, Dominique J D Vanhoutte

¹Dow Benelux, Analytical Sciences, Terneuzen, The Netherlands. redam@dow.com

Journal of Chromatography. A

|November 1, 2011

Summary

Polymer monolithic columns with tunable macropore sizes enhance size-based separations of macromolecules. This study reveals hydrodynamic chromatography as the primary mechanism, with flow rate influencing the separation of high-molecular-weight polymers.

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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

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Published on: September 21, 2011

Area of Science:

Polymer Chemistry
Chromatography
Materials Science

Background:

Size-based separation of macromolecules is crucial in various scientific fields.
Tailoring separation selectivity requires precise control over chromatographic media properties.
Polymer monoliths offer tunable pore structures for chromatographic applications.

Purpose of the Study:

To investigate the influence of macropore size in polymer monolithic columns on macromolecule separation selectivity.
To elucidate the separation mechanisms, including hydrodynamic chromatography (HDC) and size-exclusion chromatography (SEC).
To explore the impact of flow rate on the elution behavior of high-molecular-weight polymers in confined channels.

Main Methods:

Preparation of in situ polymer monolithic columns with controlled macropore sizes (75 nm to 1.2 μm).
Characterization of separation mechanisms using synthetic polymers and polystyrene standards.
Analysis of flow-rate dependent elution behavior using Deborah numbers for high-molecular-weight polymers.

Main Results:

Hydrodynamic chromatography (HDC) was identified as the dominant separation mechanism in flow-through pores.
Calibration curves for synthetic polymers aligned with HDC elution behavior for analyte-to-pore aspect ratios (λ) up to 0.2.
Deviations for small polystyrene polymers (M(r)<20 kDa) suggested a combined HDC-SEC mechanism at λ<0.02.
Separation of high-molecular-weight polymers (>300,000 Da) showed strong flow-rate dependence, attributed to shear-induced forces.

Conclusions:

Tunable macropore sizes in polymer monoliths effectively control selectivity for macromolecule separations.
The study provides insights into the interplay of HDC and SEC mechanisms based on pore size and polymer characteristics.
Flow-rate dependent elution of large polymers highlights non-classical chromatographic behaviors, potentially driven by shear forces.