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

Falck, David; Oosthoek-de Vries, Anna; Kolkman, Ard; Lingeman, Henk; Honing, Maarten; Wijmenga, Sybren; Kentgens, Arno; Niessen, Wilfried
August 2013
Analytical & Bioanalytical Chemistry;Aug2013, Vol. 405 Issue 21, p6711
Academic Journal
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.


Related Articles

  • Solution structure of a pleckstrin-homology domain. Yoon, Ho Sup; Hajduk, Philip J. // Nature;6/23/1994, Vol. 369 Issue 6482, p672 

    Presents the solution structure of the N-terminal pleckstrin-homology domain of pleckstrin determined using heteronuclear three-dimensional nuclear magnetic resonance spectroscopy. Pleckstrin as major protein kinase C substrate of platelets; Similarity of domain topology to that of the...

  • In situ NMR spectroelectrochemistry for the structure elucidation of unstable intermediate metabolites. Bussy, Ugo; Giraudeau, Patrick; Silvestre, Virginie; Jaunet-Lahary, Titouan; Ferchaud-Roucher, Véronique; Krempf, Michel; Akoka, Serge; Tea, Illa; Boujtita, Mohammed // Analytical & Bioanalytical Chemistry;Jul2013, Vol. 405 Issue 17, p5817 

    In situ NMR spectroelectrochemistry is presented in this study as a useful hybrid technique for the chemical structure elucidation of unstable intermediate species. An experimental setting was designed to follow the reaction in real time during the experimental electrochemical process. The...

  • Could smaller really be better? Current and future trends in high-resolution microcoil NMR spectroscopy. Jones, Christopher; Larive, Cynthia // Analytical & Bioanalytical Chemistry;Feb2012, Vol. 402 Issue 1, p61 

    NMR is an invaluable analytical technique that provides structural and chemical information about a molecule without destroying the sample. However, NMR suffers from an inherent lack of sensitivity compared to other popular analytical techniques. This trends article focuses on strategies to...

  • Mixed-valence properties of a dinuclear ruthenium complex bridged by bis(phenylcyanamido)tetrazine. Habibagahi, Fatemeh; Crutchley, Robert J. // Canadian Journal of Chemistry;Nov2014, Vol. 92 Issue 11, p1081 

    The novel bridging ligand 3,6-bis(phenylcyanamido)-1,2,4,5-tetrazine (tdpcH2) and its dinuclear complex [{Ru(ttpy)(bpy)}2(μ-tdpc)][PF6]2 were prepared and characterized by elemental analysis and 1H NMR spectroscopy. Cyclic voltammetry and vis-NIR and IR spectroelectrochemistry of...

  • Metabolic Profiling of the Rat Liver After Chronic Ingestion of Alpha-Naphthylisothiocyanate Using In Vivo and Ex Vivo Magnetic Resonance Spectroscopy. Solanky, Bhavana S.; Sanchez-Canon, Gina J.; Cobbold, Jeremy F. L.; Taylor-Robinson, Simon D.; Bell, Jimmy D.; Scudamore, Cheryl L.; Ross, Eleanor; Holder, Julie C.; So, Po-Wah; Cox, I. Jane // Toxicological Sciences;Apr2012, Vol. 126 Issue 2, p306 

    Editor's Hightlight: This study demonstrated that hepatobiliary toxicity could be characterized in a living animal noninvasively by 31P magnetic resonance spectroscopy. These results demonstrate the feasibility of following toxicity biomarkers longitudinally in the same animal, resulting in...

  • Feasibility study for the rapid screening of target molecules using translational diffusion coefficients: diffusion-ordered NMR spectroscopy of biological toxins. Henderson, Terry J. // Analytical & Bioanalytical Chemistry;Feb2010, Vol. 396 Issue 4, p1465 

    A panel of 15 biological toxins ranging between ~60–28,000 g/mol was used to evaluate the feasibility of screening aqueous samples for toxin analytes based on their translational diffusion coefficients, Dt. Toxin Dt values were measured by pulsed-field gradient 1H NMR spectroscopy using a...

  • Phosphorylation of histone H3 Ser10 establishes a hierarchy for subsequent intramolecular modification events. Liokatis, Stamatios; Stützer, Alexandra; Elsässer, Simon J; Theillet, Francois-Xavier; Klingberg, Rebecca; van Rossum, Barth; Schwarzer, Dirk; Allis, C David; Fischle, Wolfgang; Selenko, Philipp // Nature Structural & Molecular Biology;Jul2012, Vol. 19 Issue 8, p819 

    Phosphorylation of Ser10 of histone H3 regulates chromosome condensation and transcriptional activity. Using time-resolved, high-resolution NMR spectroscopy, we demonstrate that histone H3 Ser10 phosphorylation inhibits checkpoint kinase 1 (Chk1)- and protein kinase C (PKC)-mediated modification...

  • Structure determination of human Lck unique and SH3 domains by nuclear magnetic resonance spectroscopy. Briese, Lars; Willbold, Dieter // BMC Structural Biology;2003, Vol. 3, p3 

    Background: Protein tyrosine kinases are involved in signal transduction pathways that regulate cell growth, differentiation, activation and transformation. Human lymphocyte specific kinase (Lck) is a 56 kDa protein involved in T-cell- and IL2-receptor signaling. Three-dimensional structures are...

  • The effect of PKA-phosphorylation on the structure of inhibitor-1 studied by NMR spectroscopy. Yi-Choang Huang; Yi-Chen Chen; Huey-Jen Tsay; Chia-lin Chyan; Chun-Yu Chen; Hsien-bin Huang; Ta-Hsien Lin // Journal of Biochemistry;Feb2010, Vol. 147 Issue 2, p273 

    Inhibitor-1 is an acid- and heat-stable protein. It can be turned into a potent inhibitor of protein phosphatase-1 (PP1) after phosphorylation at Thr35 by c-AMP-dependent protein kinase (PKA). Although it has been known that pre-phosphorylation is essential for inhibition of PP1, the...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics