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EC–SPE–stripline-NMR analysis of reactive products: a feasibility study

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Abstract

Flow-through electrochemical conversion (EC) of drug-like molecules was hyphenated to miniaturized nuclear magnetic resonance spectroscopy (NMR) via on-line solid-phase extraction (SPE). After EC of the prominent p38α mitogen-activated protein kinase inhibitor BIRB796 into its reactive products, the SPE step provided preconcentration of the EC products and solvent exchange for NMR analysis. The acquisition of NMR spectra of the mass-limited samples was achieved in a stripline probe with a detection volume of 150 nL offering superior mass sensitivity. This hyphenated EC–SPE–stripline-NMR setup enabled the detection of the reactive products using only minute amounts of substrate. Furthermore, the integration of conversion and detection into one flow setup counteracts incorrect assessments caused by the degradation of reactive products. However, apparent interferences of the NMR magnetic field with the EC, leading to a low product yield, so far demanded relatively long signal averaging. A critical assessment of what is and what is not (yet) possible with this approach is presented, for example in terms of structure elucidation and the estimation of concentrations. Additionally, promising routes for further improvement of EC–SPE–stripline-NMR are discussed.

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Acknowledgments

This work was performed partly within the framework of the Dutch Top Institute Pharma project D2-102 (Metabolic stability assessment as new tool in the Hit-to-Lead selection process and the generation of new lead compound libraries) and partly within the research program ACTS—Process on a Chip (PoaC), financed by the Netherlands Organisation for Scientific Research (NWO). Agnieszka Kraj, Hendrik-Jan Brouwer and Jean-Pierre Chervet (Antec, Zoeterwoude, The Netherlands) are acknowledged for their support of the electrochemistry part of the project. Hans Janssen, Jan van Os, and Jan van Bentum (Radboud University Nijmegen, The Netherlands) are credited for technical and organizational support with regard to setup and ongoing development of the stripline probe. Additionally, Roald Tiggelaar, Jacob Bart and Han Gardeniers (Twente University/Mesoscale Chemical Systems, Enschede, The Netherlands) are acknowledged for their support of the stripline-NMR chip fabrication. Frans J.J. de Kanter and Andreas W. Ehlers are thanked for their input concerning the conventional NMR measurements.

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Authors and Affiliations

  1. AIMMS Division of BioMolecular Analysis, VU University Amsterdam, De Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands

    David Falck, Henk Lingeman, Maarten Honing & Wilfried M. A. Niessen

  2. Institute for Molecules and Materials, Radboud University Nijmegen, postvak 51, P.O. Box 9010, 6500 GL, Nijmegen, The Netherlands

    Anna J. Oosthoek-de Vries, Ard Kolkman, Sybren S. Wijmenga & Arno P. M. Kentgens

  3. DSM Resolve, Urmonderbaan 22, 6160 MD, Geleen, The Netherlands

    Ard Kolkman & Maarten Honing

  4. hyphen MassSpec, de Wetstraat 8, 2332 XT, Leiden, The Netherlands

    Wilfried M. A. Niessen

Authors
  1. David Falck
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  2. Anna J. Oosthoek-de Vries
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  3. Ard Kolkman
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  4. Henk Lingeman
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  5. Maarten Honing
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  6. Sybren S. Wijmenga
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  7. Arno P. M. Kentgens
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  8. Wilfried M. A. Niessen
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Correspondence to Wilfried M. A. Niessen.

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Falck, D., Oosthoek-de Vries, A.J., Kolkman, A. et al. EC–SPE–stripline-NMR analysis of reactive products: a feasibility study. Anal Bioanal Chem 405, 6711–6720 (2013). https://doi.org/10.1007/s00216-013-7158-9

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  • DOI: https://doi.org/10.1007/s00216-013-7158-9

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