Chem 3052 Spectroscopy

CHAPTER 7

Nuclear Magnetic Resonance Spectroscopy (NMR)

Outline:

7.1 Basic principle of NMR

7.2 NMR spectrometers
7.3 Proton NMR
7.4 C–13 NMR
7.5 Interpretation of NMR spectra
1.1. Nuclear Magnetic Resonance Spectroscopy (NMR)

Nuclear magnetic resonance (NMR) spectroscopy is based on the measurement of absorption of

electromagnetic radiation in the radio-frequency region of roughly 4 to 900 MHz. In contrast to UV-
visible, and IR absorption, nuclei of atoms rather than outer electrons are involved in the absorption
process. Furthermore, to cause nuclei to develop the energy states required for absorption to occur, it is
necessary to place the analyte in an intense magnetic field. NMR spectroscopy is one of the most
powerful tools available to chemists and biochemists for elucidating the structure of chemical species.
The technique is also useful for the quantitative determination of absorbing species. The theoretical basis

Two general types of NMR spectrometers are currently in use, continues wave (CW) and pulsed, or
Fourier transform (FT-NMR), spectrometers. All early studies were carried out with CW instruments. In
about 1970, however, FT-NMR spectrometers became available commercially, and now this type of
instrument dominates the market. In both types of instruments, the sample is positioned in a powerful
magnetic field that has strength of several tesla. CW spectrometers are similar in principle to optical
absorption instruments in that an absorption signal is monitored as the frequency of the source is slowly
scanned. In some instruments, the frequency of the source is held constant while the strength of the field
is scanned. In pulsed instruments, the sample is irradiated with periodic pulses of RF energy that are
directed through the sample at right angles to the magnetic field. These excitation pulses elicit a time-
domain signal that decays in the interval between pulses. This signal is then converted to a frequency-
domain signal by using a Fourier transformation to give a spectrum similar to that obtained by using a
CW instrument.

Nearly all NMR instruments produced today are of the FT type, and the use of CW instruments is largely
limited to special routine applications, such as the determination of the extent of hydrogenation in
petroleum process streams and the determination of water in oils, food products, and agricultural
materials. Despite this predominance of pulsed instruments in the marketplace, we find it convenient to

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base our initial development of NMR theory on CW experiments and move from there to a discussion of
pulsed NMR measurements.

Instrument manufacturers market two general types of NMR spectrometers: wide-line spectrometers and
high-resolution spectrometers. Wide-line instruments have magnets with strengths of a few tenths of a
tesla and are considerable simpler and less expensive than are high-resolution instruments. High-
resolution instruments are equipped with magnets with strengths that range from 1.4 to 23 T. which
correspond to proton frequencies of 60 to 1000 MHz (1 GHZ). Before about 1970, high-resolution NMR
spectrometers were all of the CW type that used permanent magnets or electromagnets to supply the
magnetic field. This type of instrument has been largely replaced by Fourier transform spectrometers
equipped with superconducting magnets to provide the magnetic field. Computers are an integral part of
modern NMR instruments; they digitize and store the signal, perform Fourier transformation of the FID to
provide the frequency-domain signal, and deliver many other data-treatment and instrument-control
functions. A major reason why Fourier transform instruments have become so popular is that they permit
efficient signal averaging and thus yield greatly enhanced sensitivity. Because of this greater sensitivity,
routine applications of NMR to naturally occurring carbon-13, to protons in microgram quantities, and to
other nuclei, such as fluorine, phosphorus, and silicon have become widespread. High-resolution NMR
spectrometers are expensive

Components of Fourier Transform Spectrometers

The central component of the instrument is a highly stable magnet in which the sample is placed. The
sample is surrounded by a transmitter-receiver coil. RF radiation is produced by a crystal-controlled
frequency synthesizer having an output carrier frequency of Vc. This signal passes into a pulse switch and
power amplifier, which creates an intense and reproducible pulse of RF current in the transmitter coil.
The resulting RF radiation impinges on the sample, contained inside the coil. The length, amplitude,
shape, and phase of the pulse are selected by the operator; entered into the console; and controlled by the
computer. The resulting FID signal is picked up by the same coil, which now serves as a receiver. The
signal is then amplified and transmitted to a phase-sensitive detector. The detector circuitry produces the
difference between the nuclear signals Vn and the crystal oscillator output Vc which leads to the low-
frequency, time-domain signal shown on the right of the figure. This signal is digitized and collected in
the memory of the computer for analysis by a Fourier transform program and other data-analysis
software. The output from this program is plotted on a display device, giving a frequency-domain
spectrum.

Magnets

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The heart of all NMR instruments is the magnet. The sensitivity and resolution of spectrometers are both
critically dependent on the strength and quality of their magnets. Because both sensitivity and resolution
increase with increasing field strength, it is advantageous to operate at the highest possible field strength.
In addition, the field must be highly homogeneous and reproducible. These requirements ensure that the
magnet is by far the most expensive component of an NMR spectrometer. Spectrometer magnets of three
types have been used in NMR spectrometers: permanent magnets, conventional electromagnets, and
superconducting solenoids. At present, conventional electromagnets are seldom incorporated into NMR
instruments. Permanent magnets with field strengths of 0.7, 1.4, and 2.1 T have been used in commercial
CW instruments; corresponding oscillator frequencies for proton studies are 30, 60, and 90 MHz.
Permanent magnets are highly temperature sensitive and require extensive thermo-stating and shielding as
a consequence. Because of field-drift problems, permanent magnets are not ideal for extended periods of
data accumulation, such as are often employed in Fourier transform experiments.

Superconducting magnets are used in most modern high-resolution instruments. These magnets attain
fields as large as 23 T, corresponding to a proton frequency of 1 GHz. To superconduct, the solenoid,
wound from superconducting niobium-tin or niobium-titanium wire, is bathed in liquid helium at a
temperature of 4 K. The helium Dewar flask is held in an outer liquid-nitrogen Dewar. Most
superconducting magnet systems must be filled with liquid nitrogen about once a week and with liquid
helium every few months. The advantages of superconducting solenoids, in addition to their high field
strengths, are their high stability, low operating cost, and simplicity and small size compared with an
electromagnet.

The performance specifications for a spectrometer magnet arc stringent. The field produced must be
homogeneous to a few parts per billion within the sample area and must be stable to a similar degree for
the length of time required to acquire the sample data. Unfortunately, the inherent stability of most
magnets is considerably lower than this figure, with variations as large as one part in 10 7 being observed
over a period of 1 hour. Several measures are employed in modern NMR instruments to compensate for
both drift and field in homogeneity.

Locking the Magnetic Field

To offset the effect of field fluctuations, a field-frequency lock system is employed in commercial NMR
instruments. In these systems, a reference nucleus is continuously irradiated and monitored at a frequency
corresponding to its resonance maximum at the rated field strength of the magnet. Changes in the
intensity of the reference signal control a feedback circuit, whose output is fed into coils in the magnetic
gap in such a way as to correct for the drift. Recall that for a given type of nucleus, the ratio between the
field strength and resonance frequencies is a constant regardless of the nucleus involved (Equation 19-5).

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Thus, the drift correction for the reference signal is applicable to the signals for all nuclei in the sample
area. In modern spectrometers, the reference signal is provided by the deuterium in the solvent, and a
second transmitter coil tuned to the frequency for deuterium monitors the reference. Most modern
superconducting magnets are sufficiently stable that spectra can be obtained in an unlocked mode for a
period of 1 to 20 min.

Shimming

Shim coils are pairs of wire loops through which carefully controlled currents are passed, producing small
magnetic fields that compensate for in homogeneities in the primary magnetic field. In contemporary
instruments, the shim controls are under computer control with several algorithms available for
optimizing field homogeneity. Ordinarily, shimming must be carried out each time a new sample is
introduced into the spectrometer.

Sample Spinning

The effects of field in homogeneities are also counteracted by spinning the sample along its longitudinal
axis. Spinning is accomplished by means of a small plastic turbine that slips over the sample tube. A
stream of air drives the turbine at a rate of 20 to 50 revolutions per second. If this frequency is much
greater than the frequency spread caused by magnetic in homogeneities, the nuclei experience an
averaged environment that causes apparent frequency dispersions to collapse toward zero. A minor
disadvantage of spinning is that the magnetic field is modulated at the spinning frequency, which may
lead to side hands, or spinning side hands, on each side of absorption bands.

A key component of an NMR spectrometer is the sample probe, which serves multiple functions. It holds
the sample in a fixed position in the magnetic field, contains an air turbine to spin the sample, and houses
the coil or coils that permit excitation and detection of the NMR signal. In addition, the probe ordinarily
contains two other transmitter coils, one for locking and the other for decoupling experiments. Finally,
most probes have variable temperature capability. The usual NMR sample cell consists of a 5-mm
(outside diameter) glass tube containing 500-650 µL of liquid. Micro-tubes for smaller sample volumes
and larger tunes for special needs are also available.

Transmitter-Receiver Coils

The adjustable probes in modern NMR spectrometers contain transmitter and receiver coils. The receiver
coils are arranged according to the experiment purpose. For observing protons, the prone usually has an
inner coil for lH detection and an outer coil, called the X-nucleus coil, for detecting nuclei such as 13C or
15
N. The inner coil is generally the closest to the sample to maximize sensitivity. For experiments to
detect nuclei other than protons, the X-nucleus coil is placed on the inside and the 1H coil on the outside.

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Usually, proton spectra have such good signal-to-noise ratios that probe tuning is not extremely critical.
When the X-nucleus coil is the primary observation coil, however, tuning can be very important.

The Pulse Generator:

RF generators and frequency synthesizers produce a signal of essentially a single frequency. To generate
Fourier transform spectra, however, the sample must be irradiated with a range of frequencies sufficiently
great to excite nuclei with different resonance frequencies. Fortunately, a sufficiently short pulse of
radiation provides a relatively broad band of frequencies centered on the frequency of the oscillator. The
frequency range of this band is about l/(4 ) Hz, where  is the length in seconds of each pulse. Thus, a 1-
µs pulse from a 100-MHz oscillator produces a frequency range of 100-MHz ± 125-kHz. A rectangular
waveform is synthesized from a series of sine or cosine waves differing from one another by small
frequency increments. Conversely, Fourier analysis of a square waveform reveals that it consists of a
broad range of frequency components. The narrower the square wave, the broader the range of component
frequencies. Thus, a narrow pulse generated by rapidly switching an RF oscillator consists of a band of
frequencies capable of exciting all nuclei whose resonances occur in the vicinity of the frequency of the
oscillator.

The typical pulse generator consists of three parts: a frequency synthesizer, a gate to switch the pulse on
and off at appropriate times, and a power amplifier to amplify the pulse to about 50 to 100 W.

The Receiver System:

Voltages generated by the current in the detector coil are in the nanovolt-to-microvolt range and thus
must be amplified to a range of about 0 to 10 V before the signal can be further processed and digitized.
The first stage of amplification generally takes place in a preamplifier, which is mounted in the probe so
that it is as close to the receiver coil as possible to minimize the effects of noise from other parts of the
instrument. Further amplification then is carried out in an external RF amplifier.

The Detector and Data-Processing System

In the detector system, the high-frequency radio signal is first converted to an audio-frequency signal,
which is much easier to digitize. The signal from the RF amplifier can be thought of as being made up of
two components: a carrier signal, which has the frequency of the oscillator producing it, and a
superimposed NMR signal from the analyte. The analyte signal differs in frequency from that of the
carrier by a few parts per million. For example, the chemical shifts in a proton spectrum typically
encompass a range of 10 ppm. Thus, the proton NMR data generated by a 500-MHz spectrometer would
lie in the frequency range of 500,000,000 to 500,005,000 Hz. To digitize such high-frequency signals and
extract the small differences attributed to the sample in the digital domain is not practical; therefore, the

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carrier frequency vc is subtracted electronically from the analyte signal frequency vn in the analog domain.
In our example, this process yields a difference signal (vn ~ vc) that lies in the audio-frequency range of 0
to 5000 Hz. This process is identical to the separation of audio signals from an RF carrier signal in home
radios.

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