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Mobile-Phase Contributions to Analyte Retention and Selectivity in Reversed-Phase Liquid Chromatography: 2.
Andreas Steinhoff1, Alexandra Höltzel1, Ulrich Tallarek1
1Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, Marburg 35032, Germany.
The Journal of Physical Chemistry. B
|June 16, 2025
Summary
Analyte retention in chromatography is governed by solute polarity and mobile phase composition. Molecular dynamics simulations reveal that interactions with the stationary phase and mobile phase dictate compound separation, influencing selectivity.
Area of Science:
- Analytical Chemistry
- Physical Chemistry
- Computational Chemistry
Background:
- Reversed-phase liquid chromatography (RPLC) separates compounds based on polarity.
- Mobile phase composition (water-organic solvent, W-OS) significantly impacts analyte retention and selectivity.
- Understanding molecular interactions in RPLC is crucial for method development.
Purpose of the Study:
- To investigate the molecular-level mechanisms of analyte discrimination in RPLC.
- To explore how stationary phase structure and mobile phase composition influence analyte retention.
- To elucidate the origins of selectivity in reversed-phase separations.
Main Methods:
- Molecular dynamics simulations were employed.
- A slit-pore model of a C18 silica stationary phase was used.
- Simulations were performed with water-methanol and water-acetonitrile mobile phases.
Main Results:
- Analyte retention correlated with the number of bonded-phase contacts and penetration depth.
- Hydrophobic interactions and hydrogen-bonding requirements determined analyte behavior.
- Mobile phase-induced changes in stationary phase solvation altered analyte density and selectivity.
Conclusions:
- Analyte retention is governed by a balance of hydrophobic and hydrophilic interactions.
- Selectivity arises from solute-specific responses to mobile phase-controlled solvation.
- Molecular dynamics provides insights into the fundamental principles of RPLC separations.
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