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Related Concept Videos

Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
NMR Spectroscopy: Chemical Shift Overview01:15

NMR Spectroscopy: Chemical Shift Overview

The position of the absorption signal of a sample is reported relative to the position of the signal of tetramethylsilane (TMS), which is added as an internal reference while recording spectra. The difference between the absorption frequencies of the sample and TMS (in Hz) is divided by the spectrometer operating frequency (in MHz) to obtain a dimensionless quantity called the chemical shift. It is reported on the δ (delta) scale and expressed in parts per million.
For instance, the proton...
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are slanted or...
NMR Spectroscopy: Spin–Spin Coupling01:08

NMR Spectroscopy: Spin–Spin Coupling

The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved in...
NMR Spectrometers: Overview01:20

NMR Spectrometers: Overview

NMR spectrometers consist of a strong magnet, a radiofrequency transmitter, and a detector attached to a computer console for recording spectra of samples containing NMR-active nuclei. In first-generation NMR instruments called continuous-wave spectrometers, the resonance frequencies of the nuclei are determined by frequency-sweep or field-sweep methods. The magnetic field strength is fixed and the rf signal is swept in the former, while the radiofrequency signal is fixed and the magnetic field...
NMR Spectrometers: Resolution and Error Correction01:14

NMR Spectrometers: Resolution and Error Correction

When magnetic nuclei in a sample achieve resonance and undergo relaxation, the signal detected in NMR is an approximately exponential free induction decay. Fourier transform of an exponential decay yields a Lorentzian peak in the frequency domain. Lorentzian peaks in an NMR spectrum are defined by their amplitude, full width at half maximum, and position, where the peak width is governed by the spin-spin relaxation time alone. In real experiments, however, the applied magnetic field is rendered...

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Related Experiment Video

Updated: May 13, 2026

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins
09:25

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins

Published on: November 1, 2024

SedNMR: on the edge between solution and solid-state NMR.

Ivano Bertini1, Claudio Luchinat, Giacomo Parigi

  • 1CERM, University of Florence , Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Italy, and Department of Chemistry "U. Schiff", University of Florence , via della Lastruccia 3, 50019, Sesto Fiorentino, Italy.

Accounts of Chemical Research
|March 9, 2013
PubMed
Summary
This summary is machine-generated.

Sedimented solute NMR (SedNMR) offers a novel method for preparing protein samples for solid-state NMR (SS-NMR) by inducing self-crowding. This technique simplifies sample preparation and enables the study of previously inaccessible species.

Related Experiment Videos

Last Updated: May 13, 2026

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins
09:25

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins

Published on: November 1, 2024

Area of Science:

  • Biophysics
  • Structural Biology
  • Nuclear Magnetic Resonance Spectroscopy

Background:

  • Solid-state NMR (SS-NMR) typically requires protein immobilization via crystallization, freezing, or lyophilization.
  • Protein self-crowding can also achieve sufficient molecular rotation slowdown for SS-NMR.
  • Existing methods for self-crowding include centrifugal fields for concentration gradients or ultracentrifugation for gel-like solutions.

Purpose of the Study:

  • To introduce sedimented solute NMR (SedNMR) as a minimally perturbing method for preparing biological samples for SS-NMR.
  • To explore SedNMR's potential for investigating species not amenable to traditional NMR methods.
  • To demonstrate SedNMR's utility in studying weakly binding molecular adducts.

Main Methods:

  • Sedimentation induced by magic angle spinning (MAS-induced, in situ) or ultracentrifugation (UC-induced, ex situ).
  • Utilizing centrifugal force to create concentrated, gel-like protein solutions or gradients.
  • Applying in situ MAS-induced sedimentation for combined solution and SS-NMR studies.

Main Results:

  • SedNMR provides a simple and minimally perturbing sample preparation method for SS-NMR.
  • Proteins remain hydrated in sedimented samples, potentially leading to better SS-NMR spectral resolution compared to frozen or lyophilized states.
  • In situ SedNMR successfully detected NMR signals of large molecular adducts with weak binding constants.
  • Selective sedimentation of heavier molecular species facilitates the study of complex formation.

Conclusions:

  • SedNMR is an effective technique for preparing samples for SS-NMR, especially when crystallization is challenging.
  • The method allows for the investigation of a broader range of biological species and complexes.
  • In situ SedNMR enables direct NMR observation of weakly interacting complexes, driving reactions further towards complex formation.