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The simplest approach to generate a linear magnetic eld gradient is to offset the z1 shimming from its optimal value. This linear gradient frequency labels each of the different analytes signals, so that they can be identi ed individually with their corresponding sample coil. Both 1D and 2D experiments have been carried out using this approach, in which spectra with and without a linear eld gradient were acquired.37,38 Several spectral analysis methods, including peak picking, spectral subtraction, and multiplication
Figure 7.11. A picture of the four-coil multiplex NMR probe, with Te on tubes attached to the sample capillaries to allow ow sample introduction. The sample volume for each coil was approximately 60 nL.
Figure 7.10. (A) Schematic illustration of the RF switching network used to switch one coil of the two-coil probe (developed by Zhang et al.) into the resonant circuit at a time. (B) COSY spectra of (top) sucrose and (bottom) adenosine triphosphate (both 20 mM in D2O) loaded into the two coils of the two-coil probe, respectively. (C) HMQC spectra acquired from (top) 15 N-labeled (50%-labeled) ammonium chloride in D2O and (bottom) unlabeled formamide, which were loaded in the two coils, respectively. (Adapted from J. Magn. Reson., 153: 254. Copyright Academic Press, 2001).
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Figure 7.12. Schematic illustration of the use of a eld gradient to differentiate signals from each sample in the multiplex NMR probe. The four coils are wired in parallel in a single resonant circuit. The single-arrowed line indicates the static magnetic eld (7.05 T). (A) A simulated spectrum of four water samples in the four coils is shown at right for the case of no applied eld gradient. All coils experience the same eld, and therefore only one resonance is detected. (B) An applied eld gradient (double-arrow line) changes the eld experienced by each sample, and thus four resonances are observed.
methods, have been used by these authors to generate subspectra of the samples in different coils. Two-dimensional spectroscopy can also be carried out using the multiplex NMR approach.38 For example, correlated spectroscopy (COSY) experiments have been performed with a linear z-gradient applied during the data acquisition period. Spectral analysis methods can also be used for postprocessing the data. For the latter process the authors suggested zeroing all the peaks within a certain bandwidth from the diagonal, with this bandwidth set to be less than the smallest J-coupling observed in the spectrum. Using 0.5-M samples of ethanol, 1-propanol, dichloroacetic acid and acetaldehyde in D2O, it was shown that the appropriate individual subspectra could be generated four at a time, and the subspectra of 1-propanol and ethanol are shown in Figure 7.13.
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0 1 2 3 4 5 6 7 8 9 10 10 9 8 7 6 5 4 3 2 1 0
0 1 2 3 4 5 6 7 8 9 10 10 9 8 7 6 5 4 3 2 1 0
Figure 7.13. Two-dimensional subspectra: (A) 1-propanol and (B) ethanol generated from zero-gradient and gradient shifted 2D COSY spectra of four coils using the subtraction method. (Reproduced with permission from Anal. Chim. Acta, 400: 297. Copyright Elsevier Press, 1999).
Advanced Field Gradient Methods The application of the small static magnetic eld gradients (most easily generated by offsetting the z1 shimming parameter) works well for spectra in which there is no signi cant spectral overlap between compounds, or for situations where the samples to be analyzed are quite similar. For example, parallel process monitoring could be accomplished using such an approach. In cases where spectra contain many resonances that overlap, it is much more dif cult to assign resonances based on frequency shifts. To solve this problem, it is preferable to use larger, pulsed eld gradients (PFGs) instead of the small static gradients to distinguish the different samples. PFGs are generated by a pair of oppositely wound, concentric gradient coils, which are situated around the RF coil in the probe. PFG coils that can generate large gradients of 10 to 60 G/cm are routinely used in commercial NMR probes, and their actuation can be easily realized during the pulse sequence. The exquisite control of RF pulses and PFGs currently available in modern NMR spectrometers allows a number of alternative, and more advanced, approaches to realize the multiple-sample analysis using the multiplex NMR probe. In particular, rapid selective excitation and chemical shift imaging (CSI)109,110 methods with the application of large PFGs on the order