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Related Concept Videos

NMR Spectroscopy: Chemical Shift Overview01:15

NMR Spectroscopy: Chemical Shift Overview

1.6K
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...
1.6K
Proton (¹H) NMR: Chemical Shift01:07

Proton (¹H) NMR: Chemical Shift

1.9K
Organic molecules primarily contain carbon and hydrogen atoms. While all the hydrogen isotopes are NMR-active, protium or hydrogen-1 is the most abundant. It has a significant energy separation between its nuclear spin states due to its large gyromagnetic ratio. As per Boltzmann's distribution, an increase in the energy separation implies a greater excess population of nuclei available for excitation, resulting in a strong NMR absorption signal.
Absorption signals of all the protium nuclei...
1.9K
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

731
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...
731

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Pure Shift Nuclear Magnetic Resonance: a New Tool for Plant Metabolomics
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Absolute Metabolite Quantification Using Pure Shift NMR: Toward Quantitative Metabolic Profiling of Aqueous

X Chen1, C Caradeuc1, A Montagne2

  • 1CNRS, Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques, Université Paris Cité, F-75006Paris, France.

Analytical Chemistry
|October 19, 2022
PubMed
Summary

This study introduces a new nuclear magnetic resonance (NMR) method for precise metabolite quantification in complex biological samples. The technique enhances accuracy and resolution, improving disease diagnosis and treatment strategies.

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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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Area of Science:

  • Metabolomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Accurate metabolite quantification using nuclear magnetic resonance (NMR) is crucial for understanding metabolic pathways and disease mechanisms.
  • Current NMR methods face limitations in sensitivity and resolution for analyzing complex biological samples.
  • Ultrahigh-resolution pure shift NMR methods offer improved interpretation of metabolite mixtures.

Purpose of the Study:

  • To develop a robust analytical protocol for absolute metabolite quantification in biological samples using NMR.
  • To address the sensitivity and resolution limitations of conventional NMR techniques.
  • To provide a quantitative and highly resolved method for metabolomics studies.

Main Methods:

  • Development of a pure shift NMR library of calibration reference spectra.
  • Utilizing the SAPPHIRE pulse sequence with solvent suppression for metabolite fingerprint fitting.
  • Validation using model mixtures and application to lymphoma extracellular medium.

Main Results:

  • The developed protocol demonstrated excellent trueness (slope values 0.93-1.02) and linearity (R^2 > 0.996).
  • The method achieved absolute metabolite quantification in a few hours, compatible with metabolomics workflows.
  • Successful application to determine metabolite concentrations in lymphoma extracellular medium.

Conclusions:

  • The pure shift NMR protocol provides accurate and highly resolved metabolite quantification.
  • This method significantly improves upon existing metabolomic protocols for analyzing biological samples.
  • The approach aids in understanding metabolic processes for improved disease diagnosis and treatment.