NMR Chromatography: Molecular Diffusion in the Presence of Pulsed Field Gradients in Analytical Chemistry Applications

DOI: 10.4236/ajac.2012.36053   PDF   HTML   XML   7,576 Downloads   13,710 Views   Citations


Nuclear magnetic resonance (NMR) spectroscopy is an excellent analytical tool for precise structural characterization of organic and bio-molecules. Though NMR is often used to characterize a single compound in solution, emerging experimental methods using pulsed-field-gradients (PFGs) allow structural dynamic characterization of several compounds, simultaneously in the same solution. NMR as a chromatography tool is often an overlooked application by analytical chemists. Therefore, in this article we introduce the concept of NMR based diffusion measurements, basic operational methods and novel applications. We expect this article to increase the overall use and appreciation of NMR-based diffusion measurements in general and in particular, in analytical chemistry applications.

Share and Cite:

K. Heisel, J. Goto and V. Krishnan, "NMR Chromatography: Molecular Diffusion in the Presence of Pulsed Field Gradients in Analytical Chemistry Applications," American Journal of Analytical Chemistry, Vol. 3 No. 6, 2012, pp. 401-409. doi: 10.4236/ajac.2012.36053.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] A. S. Altieri, D. P. Hinton and R. A. Byrd, “Association of Biomolecular Systems via Pulsed Field Gradient Nmr Self-Diffusion Measurements,” Journal of the American Chemical Society, Vol. 117, 28, 1995, pp. 7566-7567.
[2] A. J. Dingley, J. P. Mackay, B. E. Chapman, M. B. Morris, P. W. Kuchel, B. D. Hambly and G. F. King, “Measuring Protein Self-Association Using Pulsed-Field-Gradient Nmr Spectroscopy—Application to Myosin Light Chain 2,” Journal of Bio-molecular NMR, Vol. 6, No. 3, 1995, pp. 321-328. doi:10.1007/BF00197813
[3] V. V. Krishnan, “Determination of Oligomeric State of Proteins in Solution from Pulsed-Field-Gradient Self- Diffusion Coefficient Measurements. A Comparison of Experimental, Theoretical and Hard-Sphere Approximated Values,” Journal of Magnetic Resonance, Vol. 124, No. 2, 1997, pp. 468-473. doi:10.1006/jmre.1996.1082
[4] D. Jayawickrama, S. Zink, D. Vandervelde, R. I. Effiong and C. K. Larive, “Conformational Analysis of the Beta-Amyloid Peptide Fragment, Beta(12-28),” Journal of Biomolecular Structure & Dynamics, Vol. 13, No. 2, 1995, pp. 229-244. doi:10.1080/07391102.1995.10508836
[5] I. V. Nesmelova and V. D. Fedotov, “Self-Diffusion and Self-Association of Lysozyme Molecules in Solution,” Biochimica et Biophysica Acta-Protein Structure and Molecular Enzymology, Vol. 1383, No. 2, 1998, pp. 311- 316. doi:10.1016/S0167-4838(97)00224-0
[6] J. Lapham, J. P. Rife, P. B. Moore and D. M. Crothers, “Measurement of Diffusion Constants for Nucleic Acids by NMR,” Journal of Biomolecular NMR, Vol. 10, No. 3, 1997, pp. 255-262. doi:10.1023/A:1018310702909
[7] A. J. Lennon, N. R. Scott, B. E. Chapman and P. W. Kuchel, “Hemoglobin Affinity for 2,3-Bisphosphoglycerate in Solutions and Intact Erythrocytes: Studies Using Pulsed-Field Gradient Nuclear Magnetic Resonance and Monte Carlo Simulations,” Biophysical Journal, Vol. 67, No. 5, 1994, pp. 2096-109. doi:10.1016/S0006-3495(94)80693-9
[8] V. D?tsch and G. Wider, “Exchange Rates of Internal Water Molecules in Proteins Measured Using Pulsed Field Gradients,” Journal of the American Chemical Society, Vol. 117, No. 22, 1995, pp. 6064-6070. doi:10.1021/ja00127a020
[9] W. S. Price, M. Nara and Y. Arata, “A Pulsed Field Gradient NMR Study of the Aggregation and Hydration of Parvalbumin,” Biophysical Chemistry, Vol. 65, No. 2-3, 1997, pp. 179-187. doi:10.1016/S0301-4622(97)00003-3
[10] J. A. Jones, D. K. Wilkins, L. J. Smith and C. M. Dobson, “Characterisation of Protein Unfolding by NMR Diffusion Measurements,” Journal of Biomolecular NMR, Vol. 10, No. 2, 1997, pp. 199-203. doi:10.1023/A:1018304117895
[11] P. J. Hajduk, E. T. Olejniczak and S. W. Fesik, “One-Dimensional Relaxation- and Diffusion-Edited NMR Methods for Screening Compounds That Bind to Macromolecules,” Journal of the American Chemical Society, Vol. 119, No. 50, 1997, pp. 12257-12261. doi:10.1021/ja9715962
[12] S. Caldarelli, “Chromatographic NMR: A Tool for the Analysis of Mixtures of Small Molecules,” Magnetic Resonance in Chemistry, Vol. 45, 2007, pp. S48-S55. doi:10.1002/mrc.2143
[13] S. Cozzolino, M. G. Sanna and M. Valentini, “Probing Interactions by Means of Pulsed Field Gradient Nuclear Magnetic Resonance Spectroscopy,” Magnetic Resonance in Chemistry, Vol. 46, 2008, pp. S16-S23. doi:10.1002/mrc.2345
[14] P. Griffiths and P. Stilbs, “NMR Self-Diffusion Studies of Polymeric Surfactants,” Current Opinion in Colloid & Interface Science, Vol. 7, No. 3-4, 2002, pp. 249-252. doi:10.1016/S1359-0294(02)00042-0
[15] P. Occhipinti and P.C. Griffiths, “Quantifying Diffusion in Mucosal Systems by Pulsed-Gradient Spin-Echo NMR,” Advanced Drug Delivery Reviews, Vol. 60, No. 15, 2008, pp. 1570-1582. doi:10.1016/j.addr.2008.08.006
[16] F. Stallmach and P. Galvosas, “Spin Echo NMR Diffusion Studies,” In: G. A. Webb, Ed., Annual Reports on NMR Spectroscopy, Vol. 61, 2007, pp. 51-131.
[17] S. Zhang, “Pivotal Steps towards Quantification of Molecular Diffusion Coefficients by NMR,” Chemphyschem, Vol. 8, No. 5, 2007, pp. 635-642. doi:10.1002/cphc.200600740
[18] W. S. Price, “Pulsed-Field Gradient Nuclear Magnetic Resonance as a Tool for Studying Translational Diffusion. 1. Basic theory,” Concepts in Magnetic Resonance, Vol. 9, No. 5, 1997, pp. 299-336. doi:10.1002/(SICI)1099-0534(1997)9:5<299::AID-CMR2>3.0.CO;2-U
[19] W. S. Price, “Pulsed-Field Gradient Nuclear Magnetic Resonance as a Tool for Studying Translational Diffusion: Part II. Experimental Aspects,” Concepts in Magnetic Resonance, Vol. 10, No. 4, 1998, pp. 197-237. doi:10.1002/(SICI)1099-0534(1998)10:4<197::AID-CMR1>3.0.CO;2-S
[20] W. S. Price, “Protein Association Studied by NMR Diffusometry,” Current Opinion in Colloid & Interface Science, Vol. 11, No. 1, 2006, pp. 19-23. doi:10.1016/j.cocis.2005.10.005
[21] W. S. Price, “NMR Studies of Translational Motion,” Cambridge Molecular Science, Cambridge University Press, Cambridge, New York, 2009, xxii, 393 p.
[22] E. L. Hahn, “Spin Echoes,” Physical Review, Vol. 80, No. 4, 1950, p. 580. doi:10.1103/PhysRev.80.580
[23] E. O. Stejskal and J. E. Tanner, “Spin Diffusion Measurements; Spin Echoes in the Presence of Time Dependent Magnetic Field Gradients,” Journal of Chemical Physics, Vol. 42, No. 2, 1965, pp. 288-292. doi:10.1063/1.1695690
[24] J. Tanner, “Use of the Stimulated Echo in NMR Diffusion Studies,” The Journal of Chemical Physics, Vol. 52, 1970, p. 2523. doi:10.1063/1.1673336
[25] S. J. Gibbs and C. S. Johnson Jr., “A PFG NMR Experiment for Accurate Diffusion and Flow Studies in the Presence of Eddy Currents,” Journal of Magnetic Resonance (1969), Vol. 93, No. 2, 1991, pp. 395-402.
[26] D. Wu, A. Chen and C. S. Johnson, “An Improved Diffusion-Ordered Spectroscopy Experiment Incorporating Bipolar-Gradient Pulses,” Journal of Magnetic Resonance, Series A, Vol. 115, No. 2, 1995, pp. 260-264. doi:10.1006/jmra.1995.1176
[27] M. D. Pelta, G. A. Morris, M. J. Stchedroff and S. J. Hammond, “A One-Shot Sequence for High-Resolution Diffusion-Ordered Spectroscopy,” Magnetic Resonance in Chemistry, Vol. 40, No. 13, 2002, pp. S147-S152. doi:10.1002/mrc.1107
[28] K. F. Morris and C. S. Johnson Jr., “Diffusion-Ordered Two-Dimensional Nuclear Magnetic Resonance Spectroscopy,” Journal of the American Chemical Society, Vol. 114, No. 8, 1992, pp. 3139-3141. doi:10.1021/ja00034a071
[29] H. Barjat, G. A. Morris and A. G. Swanson, “A Three- Dimensional DOSY-HMQC Experiment for the High- Resolution Analysis of Complex Mixtures,” Journal of Magnetic Resonance, Vol. 131, No. 1, 1998, pp. 131-138. doi:10.1006/jmre.1997.1332
[30] N. Birlirakis and E. Guittet, “A New Approach in the Use of Gradients for Size-Resolved 2D-NMR Experiments,” Journal of the American Chemical Society, Vol. 118, No. 51, 1996, pp. 13083-13084. doi:10.1021/ja9627151
[31] E. K. Gozansky and D. G. Gorenstein, “DOSY-NOESY: Diffusion-Ordered NOESY,” Journal of Magnetic Resonance, Series B, Vol. 111, No. 1, 1996, pp. 94-96. doi:10.1006/jmrb.1996.0066
[32] M. Lin and M. J. Shapiro, “Mixture Analysis in Combinatorial Chemistry. Application of Diffusion-Resolved NMR Spectroscopy,” The Journal of Organic Chemistry, Vol. 61, No. 21, 1996, pp. 7617-7619. doi:10.1021/jo961315t
[33] M. Nilsson, A. M. Gil, I. Delgadillo and G. A. Morris, “Improving Pulse Sequences for 3D Diffusion-Ordered NMR Spectroscopy: 2DJ-IDOSY,” Analytical Chemistry, Vol. 76, No. 18, 2004, pp. 5418-5422. doi:10.1021/ac049174f
[34] M. Nilsson, A. M. Gil, I. Delgadillo and G. A. Morris, “Improving Pulse Sequences for 3D DOSY: COSY- IDOSY,” Chemical Communications, No. 13, 2005, pp. 1737-1739. doi:10.1039/b415099f
[35] M. J. Stchedroff, A. M. Kenwright, G. A. Morris, M. Nilsson and R. K. Harris, “2D and 3D DOSY Methods for Studying Mixtures of Oligomeric Dimethylsiloxanes,” Physical Chemistry Chemical Physics, Vol. 6, No. 13, 2004, pp. 3221-3227. doi:10.1039/b403960b
[36] R. T. Williamson, E. L. Chapin, A. W. Carr, J. R. Gilbert, P. R. Graupner, P. Lewer, P. McKamey, J. R. Carney and W. H. Gerwick, “New Diffusion-Edited NMR Experiments to Expedite the Dereplication of Known Compounds from Natural Product Mixtures,” Organic Letters, Vol. 2, No. 3, 2000, pp. 289-292. doi:10.1021/ol991239r
[37] D. Wu, A. Chen and C. S. Johnson, “Three-Dimensional Diffusion-Ordered NMR Spectroscopy: The Homonuclear COSY-DOSY Experiment,” Journal of Magnetic Resonance, Series A, Vol. 121, No. 1, 1996, pp. 88-91. doi:10.1006/jmra.1996.0142
[38] D. Wu, A. Chen and C. S. Johnson, “Heteronuclear-Detected Diffusion-Ordered NMR Spectroscopy through Coherence Transfer,” Journal of Magnetic Resonance, Series A, Vol. 123, No. 2, 1996, pp. 215-218. doi:10.1006/jmra.1996.0239
[39] J. M. Newman and A. Jerschow, “Improvements in Complex Mixture Analysis by NMR:? DQF-COSY iDOSY,” Analytical Chemistry, Vol. 79, No. 7, 2007, pp. 2957- 2960. doi:10.1021/ac061760g
[40] S. Viel and S. Caldarelli, “Improved 3D DOSY-TOCSY Experiment for Mixture Analysis,” Chemical Communications, No. 17, 2008, pp. 2013-2015. doi:10.1039/b802789g
[41] J. Sitkowski, E. B. Bednarek, W. Bocian and L. Kozerski, “Assessment of Oversulfated Chondroitin Sulfate in Low Molecular Weight and Unfractioned Heparins Diffusion Ordered Nuclear Magnetic Resonance Spectroscopy Method,” Journal of Medicinal Chemistry, Vol. 51, No. 24, 2008, pp. 7663-7665. doi:10.1021/jm801198b
[42] S. Trefi, C. Routaboul, S. Hamieh, V. Gilard, M. Malet-Martino and R. Martino, “Analysis of Illegally Manufactured Formulations of Tadalafil (Cialis?) by 1H NMR, 2D DOSY 1H NMR and Raman Spectroscopy,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 47, No. 1, 2008, pp. 103-113. doi:10.1016/j.jpba.2007.12.033
[43] S. Balayssac, V. Gilard, M. A. Delsuc and M. Malet-Martino, “DOSY NMR, a New Tool for Fake Drug Analyses,” Spectroscopy Europe, Vol. 21, No. 3, 2009, pp. 10-14.
[44] W. Bocian, R. Kaw?cki, E. Bednarek, J. Sitkowski, M. P. Williamson, P. E. Hansen and L. Kozerski, “Binding of Topotecan to a Nicked DNA Oligomer in Solution,” Chemistry—A European Journal, Vol. 14, No. 9, 2008, pp. 2788-2794. doi:10.1002/chem.200700732
[45] J. S. Gounarides, A. Chen and M. J. Shapiro, “Nuclear Magnetic Resonance Chromatography: Applications of Pulse Field Gradient Diffusion NMR to Mixture Analysis and Li-gand-Receptor Interactions,” Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 725, No. 1, 1999, pp. 79-90. doi:10.1016/S0378-4347(98)00512-X
[46] C. S. Johnson Jr., “Effects of Chemical Exchange in Diffusion-Ordered 2D NMR Spectra,” Journal of Magnetic Resonance, Series A, Vol. 102, No. 2, 1993, pp. 214-218. doi:10.1006/jmra.1993.1093
[47] W. S. Price, “Recent Advances in NMR Diffusion Techniques for Studying Drug Binding,” Australian Journal of Chemistry, Vol. 56, No. 9, 2003, pp. 855-860. doi:10.1071/CH03128
[48] K. F. Morris, P. Stilbs and C. S. Johnson, “Analysis of Mixtures Based on Molecular Size and Hydrophobicity by Means of Diffusion-Ordered 2D NMR,” Analytical Chemistry, Vol. 66, No. 2, 1994, pp. 211-215. doi:10.1021/ac00074a006
[49] R. Evans, S. Haiber, M. Nilsson and G. A. Morris, “Isomer Resolution by Micelle-Assisted Diffusion-Ordered Spectroscopy,” Analytical Chemistry, Vol. 81, No. 11, 2009, pp. 4548-4550. doi:10.1021/ac9005777
[50] S. Viel, F. Ziarelli and S. Caldarelli, “Enhanced Diffusion-Edited NMR Spectroscopy of Mixtures Using Chromatographic Stationary Phases,” Proceedings of the National Academy of Sciences, Vol. 100, No. 17, 2003, pp. 9696-9698. doi:10.1073/pnas.1533419100
[51] M. E. Zielinski and K. F. Morris, “Using Perdeuterated Surfactant Micelles to Resolve Mixture Components in Diffusion-Ordered NMR Spectroscopy,” Magnetic Resonance in Chemistry, Vol. 47, No. 1, 2009, pp. 53-56. doi:10.1002/mrc.2348
[52] K. F. Morris, B. A. Becker, B. C. Valle, I. M. Warner and C. K. Larive, “Use of NMR Binding Interaction Mapping Techniques to Examine Interactions of Chiral Molecules with Molecular Micelles,” The Journal of Physical Chemistry B, Vol. 110, No. 35, 2006, pp. 17359-17369. doi:10.1021/jp0627224
[53] C. F. Tormena, R. Evans, S. Haiber, M. Nilsson and G. A. Morris, “Matrix-Assisted Diffusion-Ordered Spectroscopy: Mixture Resolution by NMR Using SDS Micelles,” Magnetic Resonance in Chemistry, Vol. 48, No. 7, 2010, pp. 550-553. doi:10.1002/mrc.2621
[54] F. Asaro and N. Savko, “Resolution of a Nonionic Surfactant Oligomeric Mixture by Means of DOSY with Inverse Micelle Assistance,” Magnetic Resonance in Chemistry, Vol. 49, No. 4, 2011, pp. 195-198. doi:10.1002/mrc.2732
[55] Y. Cohen, L. Avram and L. Frish, “Diffusion NMR Spectroscopy in Supramolecular and Combinatorial Chemistry: An Old Parameter—New Insights,” Angewandte Chemie International Edition, Vol. 44, No. 4, 2005, pp. 520-554. doi:10.1002/anie.200300637

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.