NATURAL ABUNDANCE NITROGEN-15 NMR BY ENHANCED HETERONUCLEAR SPECTROSCOPY

NATURAL ABUNDANCE NITROGEN-15 NMR BY ENHANCED HETERONUCLEAR SPECTROSCOPY

1 January 1980 | Geoffrey BODENHAUSEN and David J. RUBEN
A two-dimensional Fourier transform technique involving a double transfer of polarization significantly improves the detection of NMR spectra of less sensitive nuclei coupled to protons. This method allows for the acquisition of natural abundance $ ^{15} $N spectra in small sample volumes using a commercial spectrometer. The technique involves generating proton magnetization, transferring coherence to nitrogen transitions, and then transferring it back to proton transitions to enable high-frequency detection. The pulse sequence includes a series of pulses that rotate magnetization and refocus signals, with delays adjusted to probe nitrogen chemical shifts. The proton decoupling of the nitrogen spectrum and the nitrogen decoupling of the proton spectrum are optional. The method is applied to a 1 M solution of 99% enriched N-acetyl valine in DMSO-d6, producing proton spectra that show nitrogen-15 chemical shifts. The technique suppresses proton signals not originating from $ ^{15} $N transitions, allowing for the study of materials not abundantly available. The experiment is suitable for studies in deuterated solvents and non-exchanging NH groups of proteins in D2O. The method enhances sensitivity by transferring magnetization from protons to nitrogen, resulting in a seventeen-fold enhancement in N-acetyl valine. The technique involves a ten-pulse sequence that generates a two-dimensional spectrum, with the $ F_1 $ domain corresponding to $ ^{15} $N spectra and the $ F_2 $ domain to proton spectra. The method is effective in reducing the nuclear Overhauser effect and provides accurate measurements of $ ^{15} $N chemical shifts. The experiments were conducted using a Bruker 270 MHz spectrometer, with the probe tuned for proton observation and the decoupler coil retuned for $ ^{15} $N. The sensitivity advantage is determined by the factor $ (\gamma_H/\gamma_N)^{5/2} $, although the method is time-consuming for high-resolution $ ^{15} $N spectra. The technique is suitable for studying materials with low natural abundance of $ ^{15} $N.A two-dimensional Fourier transform technique involving a double transfer of polarization significantly improves the detection of NMR spectra of less sensitive nuclei coupled to protons. This method allows for the acquisition of natural abundance $ ^{15} $N spectra in small sample volumes using a commercial spectrometer. The technique involves generating proton magnetization, transferring coherence to nitrogen transitions, and then transferring it back to proton transitions to enable high-frequency detection. The pulse sequence includes a series of pulses that rotate magnetization and refocus signals, with delays adjusted to probe nitrogen chemical shifts. The proton decoupling of the nitrogen spectrum and the nitrogen decoupling of the proton spectrum are optional. The method is applied to a 1 M solution of 99% enriched N-acetyl valine in DMSO-d6, producing proton spectra that show nitrogen-15 chemical shifts. The technique suppresses proton signals not originating from $ ^{15} $N transitions, allowing for the study of materials not abundantly available. The experiment is suitable for studies in deuterated solvents and non-exchanging NH groups of proteins in D2O. The method enhances sensitivity by transferring magnetization from protons to nitrogen, resulting in a seventeen-fold enhancement in N-acetyl valine. The technique involves a ten-pulse sequence that generates a two-dimensional spectrum, with the $ F_1 $ domain corresponding to $ ^{15} $N spectra and the $ F_2 $ domain to proton spectra. The method is effective in reducing the nuclear Overhauser effect and provides accurate measurements of $ ^{15} $N chemical shifts. The experiments were conducted using a Bruker 270 MHz spectrometer, with the probe tuned for proton observation and the decoupler coil retuned for $ ^{15} $N. The sensitivity advantage is determined by the factor $ (\gamma_H/\gamma_N)^{5/2} $, although the method is time-consuming for high-resolution $ ^{15} $N spectra. The technique is suitable for studying materials with low natural abundance of $ ^{15} $N.
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