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Non-particulate (continuous bed or monolithic) restricted-access reversed-phase media for sample clean-up and separation by capillary-format liquid chromatography

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Abstract

Restricted-access reversed-phase non-particulate (continuous bed or monolithic) stationary phases of different hydrophobicity synthesized in 100 μm i.d. fused silica capillaries have been evaluated. A specific property of restricted-access media (RAM) is that they interact with small analytes and exclude big molecules, e.g. proteins, from access to the active sites and adsorption on the surface. This dual property facilitates direct injection of biological fluids for drug or drug-metabolite analysis. Different RAM and RAM-precursor capillary columns were tested to assess the influence of chromatographic bed morphology on loadability. Inverse size-exclusion chromatography was used for investigation of pore structural properties of the capillary-format continuous beds. The data obtained were used to discuss the mechanism of separation of the biological samples using capillary columns and to propose a model for the topochemical architecture of the RAM investigated. Different morphology of the non-particulate reversed-phase precursors resulted in two types of RAM material shielded with hydrophilic polymer, classified as homogeneous or heterogeneous topochemistry stationary phases. Capillary columns were applied for chromatography of biological fluids. High resolution was obtained, without the need for column switching, when capillary columns operated in gradient conditions. Extensive evaluation of the chromatographic properties (hydrophobicity, efficiency, separation impedance, and loadability) of the non-particulate reversed-phase materials was performed before and after shielding with hydrophilic polymer to generate restricted-access properties. Minor changes of hydrophobicity, efficiency, or separation impedance were observed after the shielding.

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Acknowledgements

The work was supported by the Royal Swedish Academy of Sciences (Sweden).

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Authors and Affiliations

  1. Vytautas Magnus University, Vileikos str. 8, 3035, Kaunas, Lithuania

    Reda Jarmalavičienė, Olga Kornyšova & Audrius Maruška

  2. Analytical Pharmaceutical Chemistry, Uppsala University, P.O. Box 574, 751 23, Uppsala, Sweden

    Douglas Westerlund

Authors
  1. Reda Jarmalavičienė
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  2. Olga Kornyšova
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  3. Douglas Westerlund
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  4. Audrius Maruška
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Correspondence to Audrius Maruška.

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Jarmalavičienė, R., Kornyšova, O., Westerlund, D. et al. Non-particulate (continuous bed or monolithic) restricted-access reversed-phase media for sample clean-up and separation by capillary-format liquid chromatography. Anal Bioanal Chem 377, 902–908 (2003). https://doi.org/10.1007/s00216-003-2244-z

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  • DOI: https://doi.org/10.1007/s00216-003-2244-z

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