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“Pseudostationary Ion-Exchanger” Sweeping as an Online Enrichment Technique in the Determination of Nucleosides in Urine via Micellar Electrokinetic Chromatography

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Abstract

The presented study aims to develop a new online enrichment strategy [“pseudostationary ion-exchanger” (PSIE) sweeping] for the analysis of highly hydrophilic nucleosides in urine samples with a special focus on the fundamental aspects regarding the enrichment process itself. In the first method, we employ the ionic liquid (IL)-type surfactant 1-tetradecyl-3-methylimidazolium bromide (C14MImBr) as micelle forming agent under alkaline pH conditions. It is shown that maximum enrichment efficiency can be obtained by keeping the retention factors very high within the sample zone and very low within the background electrolyte (BGE) while maintaining a sufficient resolution for the studied analytes. With this method, detection limits as low as 0.1 µg mL−1 are obtained for all analytes studied. For the nucleosides, adenosine and cytidine, a second method is developed using sodium dodecyl sulfate (SDS) as micelle forming agent under acidic pH conditions. In addition, we investigate the effect of replacing ionic buffering constituents with a zwitterionic/isoelectric buffering compound (aspartic acid) with regard to separation and enrichment efficiency. With the second method, the achieved limits of detection are as low as 0.1 µg mL−1 for Ado and 0.2 µg mL–1 for Cyd. The applicability of the two complementary methods to the analysis of the nucleosides under investigation is shown for blank and spiked human urine samples after their extraction using the commercially available phenylboronate affinity gel.

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Acknowledgements

We thank the workshops of the Department of Chemistry, University of Marburg for the development of the data recording unit.

Funding

A. H. Rageh thanks the Egyptian Ministry of Higher Education and the Ministry of State for Scientific Research and the Deutscher Akademischer Austauschdienst (DAAD) for funding her PhD scholarship through German Egyptian Research Long-Term Scholarship program (GERLS).

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Authors and Affiliations

  1. Department of Chemistry, University of Marburg, Hans-Meerwein-Straße, 35032, Marburg, Germany

    Azza H. Rageh & Ute Pyell

  2. Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, University of Assiut, Assiut, 71526, Egypt

    Azza H. Rageh

Authors
  1. Azza H. Rageh
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  2. Ute Pyell
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Correspondence to Ute Pyell.

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Conflict of interest

A. H. Rageh declares that she has no conflict of interest. U. Pyell declares that she has no conflict of interest.

Ethical approval

All procedures performed in  this study involving a human participant were in accordance with the 1964 Helsinki Declaration and its later amendments. For experiments with human urine, informed consent was obtained from the volunteer. This article does not contain any studies with animals performed by any of the authors.

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Published in Chromatographia’s 50th Anniversary Commemorative Issue.

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Rageh, A.H., Pyell, U. “Pseudostationary Ion-Exchanger” Sweeping as an Online Enrichment Technique in the Determination of Nucleosides in Urine via Micellar Electrokinetic Chromatography. Chromatographia 82, 325–345 (2019). https://doi.org/10.1007/s10337-018-3570-9

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