NUCLEAR MAGNETIC RESONANCE
INTRODUCTION
➢ Nuclear magnetic resonance (NMR), selective absorption of very high-frequency radio waves by certain atomic nuclei that are
subjected to an appropriately strong stationary magnetic field. This phenomenon was first observed in 1946 by the physicists
Felix Bloch and Edward M. Purcell independently of each other. Nuclei in which at least one proton or one neutron is unpaired
act like tiny magnets, and a strong magnetic field exerts a force that causes them to process in somewhat the same way that
the axes of spinning tops trace out cone-shaped surfaces while they process in the Earth’s gravitational field. When the natural
frequency of the processing nuclear magnets corresponds to the frequency of a weak external radio wave striking the material,
energy is absorbed from the radio wave. This selective absorption, called resonance, may be produced either by tuning the
natural frequency of the nuclear magnets to that of a weak radio wave of fixed frequency or by tuning the frequency of the
weak radio wave to that of the nuclear magnets (determined by the strong constant external magnetic field). See also magnetic
resonance.
➢ Nuclear magnetic resonance is used to measure nuclear magnetic moments, the characteristic magnetic behaviour of specific
nuclei. Because these values are significantly modified by the immediate chemical environment, however, NMR measurements
provide information about the molecular structure of various solids and liquids.
➢ By the early 1980s nuclear magnetic resonance techniques had begun to be used in medicine to visualize soft tissues of the
body. This application of NMR, called magnetic resonance imaging (MRI), presented a hazard-free, non-invasive way to
generate visual images of thin slices of the body by measuring the nuclear magnetic moments of ordinary hydrogen nuclei in
the body’s water and lipids (fats). NMR images show great sensitivity in differentiating between normal tissues and diseased or
damaged ones. By the late 1980s MRI had proved superior to most other imaging techniques in providing images of the brain,
heart, liver, kidneys, spleen, pancreas, breast, and other organs. MRI provides relatively high-contrast, variable-toned images
that can show tumours, blood-starved tissues, and neural plaques resulting from multiple sclerosis. The technique presents no
known health hazards, but it cannot be used on individuals who have cardiac pacemakers or certain other metal-containing
devices implanted in their bodies.
What is NMR?
➢ Nuclear magnetic resonance spectroscopy, most commonly known as NMR spectroscopy or magnetic resonance spectroscopy
(MRS), is a spectroscopic technique to observe local magnetic fields around atomic nuclei.
➢ It is a spectroscopy technique that is based on the absorption of electromagnetic radiation in the radiofrequency region 4 to
900 MHz by nuclei of the atoms.
➢ Over the past fifty years, NMR has become the preeminent technique for determining the structure of organic compounds.
➢ Of all the spectroscopic methods, it is the only one for which a complete analysis and interpretation of the entire spectrum is
normally expected.
Nuclear Magnetic Resonance (NMR) Spectroscopy
Theory
➢ The chemical theory that underlies NMR spectroscopy depends on the intrinsic spin of the nucleus involved, described by the
quantum number S. Nuclei with a non-zero spin are always associated with a non-zero magnetic moment, where μ is the
➢ magnetic moment, S is the spin, and γ is always non-zero. It is this magnetic moment that allows for NMR to be used;
therefore, nuclei whose quantum spin is zero cannot be measured using NMR. Almost all isotopes that have both an even
number of protons and neutrons have no magnetic moment, and cannot be measured using NMR.
➢ μ= γ⋅S
➢ In the presence of an external magnetic field (B) for a nuclei with a spin I = 1/2, there are two spin states present of +1/2 and -
1/2. The difference in energy between these two states at a specific external magnetic field (Bx).
➢ where E is energy, I is the spin of the nuclei, and μ is the magnetic moment of the specific nuclei being analyzed. The difference
in energy shown is always extremely small, so for NMR strong magnetic fields are required to further separate the two energy
states. At the applied magnetic fields used for NMR, most magnetic resonance frequencies tend to fall in the radio frequency
range.
➢ E = μ⋅Bx/I
The difference in energy between two spin states over a varying magnetic field B.
➢ The reason NMR can differentiate between different elements and isotopes is due to the fact that each specific nuclide will
only absorb at a very specific frequency. This specificity means that NMR can generally detect one isotope at a time, and this
results in different types of NMR: such as 1H NMR, 13C NMR, and 31P NMR, to name only a few.
➢ The subsequent absorbed frequency of any type of nuclei is not always constant, since electrons surrounding a nucleus can
result in an effect called nuclear shielding, where the magnetic field at the nucleus is changed (usually lowered) because of the
surrounding electron environment. This differentiation of a particular nucleus based upon its electronic (chemical) environment
allows NMR be used to identify structure. Since nuclei of the same type in different electron environments will be more or less
shielded than another, the difference in their environment (as observed by a difference in the surrounding magnetic field) is
defined as the chemical shift.
Principle of Nuclear Magnetic Resonance (NMR) Spectroscopy
➢ The principle behind NMR is that many nuclei have spin and all nuclei are electrically charged. If an external magnetic field is
applied, an energy transfer is possible between the base energy to a higher energy level (generally a single energy gap).
➢ The energy transfer takes place at a wavelength that corresponds to radio frequencies and when the spin returns to its base
level, energy is emitted at the same frequency.
➢ The signal that matches this transfer is measured in many ways and processed in order to yield an NMR spectrum for the
nucleus concerned.
➢ All nuclei are electrically charged and many have spin.
➢ Transfer of energy is possible from base energy to higher energy levels when an external magnetic field is applied.
➢ The transfer of energy occurs at a wavelength that coincides with the radio frequency.
➢ Also, energy is emitted at the same frequency when the spin comes back to its base level.
➢ Therefore, by measuring the signal which matches this transfer the processing of the NMR spectrum for the concerned
nucleus is yield.
Working of Nuclear Magnetic Resonance (NMR) Spectroscopy
➢ The sample is placed in a magnetic field and the NMR signal is produced by excitation of the nuclei sample with radio waves
into nuclear magnetic resonance, which is detected with sensitive radio receivers.
➢ The intramolecular magnetic field around an atom in a molecule changes the resonance frequency, thus giving access to details
of the electronic structure of a molecule and its individual functional groups.
➢ As the fields are unique or highly characteristic to individual compounds, NMR spectroscopy is the definitive method to identify
monomolecular organic compounds.
➢ Besides identification, NMR spectroscopy provides detailed information about the structure, dynamics, reaction state, and
chemical environment of molecules.
➢ The most common types of NMR are proton and carbon-13 NMR spectroscopy, but it is applicable to any kind of sample that
contains nuclei possessing spin.
Instrumentation of Nuclear Magnetic Resonance (NMR) Spectroscopy
Sample holder
Glass tube with 8.5 cm long, 0.3 cm in diameter.
Permanent magnet
It provides a homogeneous magnetic field at 60-100 MHZ
Magnetic coils
These coils induce a magnetic field when current flows through them
Sweep generator
To produce an equal amount of magnetic field pass through the sample
Radio frequency transmitter
A radio transmitter coil transmitter that produces a short powerful pulse of radio waves
Radio frequency receiver
A radio receiver coil that detects radio frequencies emitted as nuclei relax to a lower energy level
Read out systems
A computer that analyses and records the data.
Applications of Nuclear Magnetic Resonance (NMR) Spectroscopy
➢ Spectroscopy is the study of the interaction of electromagnetic radiation with matter. NMR spectroscopy is the use of the NMR
phenomenon to study the physical, chemical, and biological properties of matter.
➢ It is an analytical chemistry technique used in quality control.
➢ It is used in research for determining the content and purity of a sample as well as its molecular structure. For example, NMR
can quantitatively analyze mixtures containing known compounds.
➢ NMR spectroscopy is routinely used by chemists to study chemical structure using simple one-dimensional techniques. Two-
dimensional techniques are used to determine the structure of more complicated molecules.
➢ These techniques are replacing x-ray crystallography for the determination of protein structure.
➢ Time domain NMR spectroscopy techniques are used to probe molecular dynamics in solution.
➢ Solid state NMR spectroscopy is used to determine the molecular structure of solids.
➢ Other scientists have developed NMR methods-of measuring diffusion coefficients.
➢ Identification of functional groups: Every functional group has characteristic signal in the NMR spectrum. By studying the
chemical shift of the compound, it is possible to identify the functional group in the compound.
➢ Structure elucidation of unknown compound: By observing the chemical shift values and splitting of signal of protons under
different environment, it is possible to place hydrogen at suitable place in the formula and hence structure of given compound
can be established.
➢ Comparison of two compounds and determination of the molecular weight: By comparing the peak of the unknown
compound with the of unknown compound, molecular weight can be determined.
➢ Distinction between cis-trans isomer and conformers: Protons have different values of chemical as well as coupling constants
in cis and trans isomer. Therefore, it is easy to identify cis and trans isomer.
Factors affecting NMR spectroscopy
Some of the factors that affect NMR Spectroscopy are given below:
• Nature of group
• Electronegativity
• Inductive effect
• Hybridization
Chemical Shift in NMR Spectroscopy
➢ A spinning charge generates a magnetic field that results in a magnetic moment proportional to the spin. In the presence of an
external magnetic field, two spin states exist; one spin up and one spin down, where one aligns with the magnetic field and the
other opposes it.
➢ Chemical shift is characterized as the difference between the resonant frequency of the spinning protons and the signal of the
reference molecule. Nuclear magnetic resonance chemical change is one of the most important properties usable for molecular
structure determination. There are also different nuclei that can be detected by NMR spectroscopy, 1H (proton), 13C (carbon
13), 15N (nitrogen 15), 19F (fluorine 19), among many more. 1H and 13C are the most widely used. The definition of 1H as it is
very descriptive of the spectroscopy of the NMR. Both the nuts have a good charge and are constantly revolving like a cloud.
Through mechanics, we learn that a charge in motion produces a magnetic field. In NMR, when we reach the radio frequency
(Rf) radiation nucleus, it causes the nucleus and its magnetic field to turn (or it causes the nuclear magnet to pulse, thus the
term NMR).
Limitations of NMR
➢ Despite all of its upsides, there are several limitations that can make NMR analysis difficult or impossible in certain situations.
One such issue is that the desired isotope of an element that is needed for NMR analysis may have little or no natural
abundance. For example, the natural abundance of 13C, the active isotope for carbon NMR, is about 11%, which works well for
analysis. However, in the case of oxygen the active isotope for NMR is 17O, which is only 0.035% naturally abundant. This
means that there are certain elements that can essentially never be measured through NMR.
➢ Another problem is that some elements have an extremely low magnetic moment, μ. The sensitivity of NMR machines is based
on the magnetic moment of the specific element, but if the magnetic moment is too low it can be very difficult to obtain an
NMR spectra with enough peak intensity to properly analyze.
NMR in biology
➢ In biology, NMR is fundamental for determining and exploring the structure of proteins, e.g., enzymes, receptors. It has been
used to elucidate the structure and function of numerous biological components. More recent discoveries include
determination of the structure of the influenza virus proton transporter and the consequent development of molecules that
block this transporter and stall infection by the virus.
➢ Similarly, determination of the structure of the CMAT receptor (an oncogene that is involved in cancer metastasis) has allowed
a variant to be engineered that has an antagonistic effect. NMR thus plays a key role in the development of vaccines and
treatments for a range of diseases, including HIV, influenza, tuberculosis and cancer, by allowing researchers to understand
how the disease-causing agents function and to identify potential drug targets.
➢ NMR also provides a powerful tool for studying conformational changes in proteins and chemical kinetic processes, which
enables the mode of action of enzymes, transporter proteins etc to be elucidated at a molecular level. For example, it was
discovered that dimerization is crucial for the ECAD protein to create adhesion between cells in multicellular organisms.
➢ Similarly, NMR is being used to characterise the transition of proteins in the brain and to investigate the influence of such
changes on neurodegenerative disease processes, such as Alzheimer's disease, Parkinson's disease, and Creutzfeldt-Jakob
disease. Ultimately, such NMR studies may again lead to the development of tests and treatments for diseases in man.
Nuclear Magnetic Resonance - Continuous-wave Nuclear Magnetic Resonance
A continuous-wave NMR instrument consists of the following units: a magnet to separate the nuclear spin energy states; at least two
radiofrequency channels, one for field/frequency stabilization and one to furnish RF irradiation energy; a sample probe containing coils for
coupling the sample with the RF field; a detector to process the NMR signals; a sweep generator for sweeping either the magnetic or RF
field through the resonance frequencies of the sample; and a recorder to display the spectrum.
The spectrum is scanned by the field-sweep method or the frequency-sweep method. In the frequency-sweep method, the magnetic field
is held constant, which keeps the nuclear spin energy levels constant, then the RF signal is swept to determine the frequencies at which
energy is absorbed. In the field sweep method, the RF signal is held constant, then the magnetic field is swept, which varies the energy
levels, to determine the magnetic field strengths that produce resonance at fixed resonance frequency.
Nuclear Magnetic Resonance - Fourier-Transform Nuclear Magnetic Resonance
Fourier-Transform NMR spectrometers use a pulse of radiofrequency radiation to cause nuclei in a magnetic field to flip into the higher-
energy alignment. The length of the RF pulse is 1-10 µs and is wide enough to simultaneously excite nuclei in all local environments. The
interval between pulses T is typically one to several seconds. During T, a time-domain RF signal called the free induction decay (FID) signal
is emitted as nuclei return to their original state.
FID can be detected with a radio-receiver coil that is perpendicular to the static magnetic field. The FID signal is digitized and stored in a
computer for data processing. Ordinary the time-domain decay signals from numerous successive pulses can be summed and averaged to
improve the signal-to-noise ratio. The result is then converted to a frequency-domain signal by a Fourier transformation. The resulting
frequency-domain output is similar to the spectrum produced by a scanning continuous-wave experiment.
References
http://chem.ch.huji.ac.il/nmr/whatisnmr/whatisnmr.html
https://www.slideshare.net/solairajananant/nmr-spectroscopy-13887430
https://www.ias.ac.in/article/fulltext/reso/009/01/0034-0049
https://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/nmr/nmr1.htm
https://en.wikipedia.org/wiki/Nuclear_magnetic_resonance_spectroscopy
