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Related Concept Videos

Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

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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...
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NMR Spectrometers: Overview01:20

NMR Spectrometers: Overview

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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...
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2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

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Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
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NMR Spectrometers: Resolution and Error Correction01:14

NMR Spectrometers: Resolution and Error Correction

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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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Quantitative Analysis01:12

Quantitative Analysis

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Quantitative analysis is a technique for measuring the amount of specific constituents in a sample. When the sample's composition is unknown, qualitative analysis is performed first to identify its components, which ensures that the correct substances are measured during the quantitative phase.
In quantitative analysis, two key measurements are made: the sample quantity and a property proportional to the amount of the analyte (the substance being analyzed). This forms the basis of the...
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NMR Spectroscopy: Chemical Shift Overview01:15

NMR Spectroscopy: Chemical Shift Overview

2.8K
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...
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NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements
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Quantitative analysis using external standards with a benchtop NMR spectrometer.

Yejin Lee1, Yevgen Matviychuk1, Daniel J Holland1

  • 1University of Canterbury, Private Bag 4800, Christchurch 8140, New Zealand.

Journal of Magnetic Resonance (San Diego, Calif. : 1997)
|October 2, 2020
PubMed
Summary
This summary is machine-generated.

Benchtop NMR quantitative analysis using external standards shows errors over 4% due to instrument limitations. Using solvent peaks as internal standards and improved spectral processing methods can enhance accuracy for aqueous samples.

Keywords:
Benchtop instrumentsExternal standardMedium-field NMRMixture analysisQuantitative NMR spectroscopy

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Area of Science:

  • Analytical Chemistry
  • Spectroscopy
  • Nuclear Magnetic Resonance (NMR)

Background:

  • Benchtop NMR spectrometers offer accessible high-resolution NMR analysis.
  • Quantitative analysis using external standards in NMR can be challenging due to instrument variability.
  • Accurate quantification is crucial for various chemical and biological applications.

Purpose of the Study:

  • To evaluate the quantitative accuracy of benchtop NMR spectrometers using external standards for aqueous samples.
  • To identify sources of error in benchtop NMR quantitative analysis.
  • To propose methods for improving the accuracy and reliability of these measurements.

Main Methods:

  • Quantitative analysis of aqueous samples (30 mM to 1.7 M) using a 43 MHz benchtop NMR spectrometer.
  • Application of the PULCON method for quantification with external standards.
  • Investigation of solvent peak integral area as an internal standard.
  • Round robin study involving manual spectral processing by multiple analysts.
  • Implementation of heuristics for manual baseline and phase correction.
  • Semi-automated quantification using the qGSD method.

Main Results:

  • The PULCON method with external standards resulted in >4% quantification error for dissimilar analytes and standards, attributed to fixed tuning/matching of the benchtop NMR.
  • Solvent peak integration proved effective as an internal standard for moderately dilute samples (<0.2 M), mitigating errors.
  • Manual spectral processing introduced significant analyst-dependent uncertainty.
  • Proposed heuristics reduced manual processing errors to ~3%.
  • Semi-automated qGSD achieved comparable integration accuracy with reduced operator dependency.

Conclusions:

  • Benchtop NMR spectrometers require careful method development for accurate quantitative analysis, especially with external standards.
  • Internal standardization using solvent peaks and refined spectral processing are key to improving accuracy.
  • Semi-automated methods like qGSD offer a more robust approach to reduce operator-induced variability in quantitative NMR.