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NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences

NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences

A pulse is a short burst of radio waves distributed over a range of frequencies that simultaneously excites all the nuclei in the sample. Upon passing a radio frequency pulse along the x-axis, the nuclei absorb energy corresponding to their Larmor frequencies and achieve resonance. This shifts the net magnetization vector from the z-axis toward the transverse plane. This angle of rotation of the magnetization vector, or the flip angle, is proportional to the duration and intensity of the pulse.

¹H NMR Signal Integration: Overview

¹H NMR Signal Integration: Overview

The intensity of a signal, which can be represented by the area under the peak, depends on the number of protons contributing to that signal. The area under each peak is shown as a vertical line called an integral, with the integral value listed under it, as seen in the proton NMR spectrum of benzyl acetate. Each integral value is divided by the smallest integral value to obtain the ratio of the number of protons producing each signal. The ratio reveals the relative number of protons and not...

Applications Of NMR In Biology

Applications Of NMR In Biology

Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
The...

NMR Spectrometers: Overview

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...

¹H NMR: Interpreting Distorted and Overlapping Signals

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...

¹³C NMR: ¹H–¹³C Decoupling

¹³C NMR: ¹H–¹³C Decoupling

The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...

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Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

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NMR quantification using an artificial signal.

Roger D Farrant¹, John C Hollerton, Sean M Lynn

¹Analytical Chemistry, GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire SG1 2NY, UK.

Magnetic Resonance in Chemistry : MRC

|August 31, 2010

Summary

We developed QUANTAS, a new software method for accurate Nuclear Magnetic Resonance (NMR) concentration measurements. This approach simplifies quantification by compensating for experimental variables, making it broadly applicable.

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Area of Science:

Analytical Chemistry
Spectroscopy
Nuclear Magnetic Resonance (NMR)

Background:

Accurate concentration determination is crucial in various scientific fields.
Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful tool for molecular analysis.
Existing NMR quantification methods can be sensitive to experimental variations.

Purpose of the Study:

To introduce QUANTAS (QUANTification by Artificial Signal), a novel software-based protocol for NMR.
To establish an absolute intensity external standard method for NMR quantification.
To develop a method that compensates for diverse experimental parameters.

Main Methods:

Developed a software-based protocol named QUANTAS.
Employed an absolute intensity external standard approach.
Demonstrated applicability for (1)H and (19)F NMR.
Incorporated compensation for experimental parameters like probe tuning, matching, and pulse length.
Utilized PULCON (PUlse Length-based CONcentration measurements) for absolute intensity corrections when necessary.

Main Results:

QUANTAS enables accurate concentration measurements by NMR.
The method compensates for variations in experimental parameters.
It allows for the comparison of heteronuclear integrals.
Demonstrated successful application in (1)H and (19)F NMR spectroscopy.
The protocol is compatible with all nuclei and modern NMR spectrometers.

Conclusions:

QUANTAS provides a robust and versatile solution for NMR quantification.
The software-based protocol simplifies and enhances the accuracy of concentration measurements.
It offers broad applicability across different nuclei and NMR hardware.