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Nuclear Overhauser Enhancement (NOE)

Irradiation of one nucleus while observing the resonance of another may result in a change in the amplitude for the
observed resonance, generally an enhancement of the signal intensity. This is known as nuclear Overhauser enhancement
(NOE). The NOE is a through space effect, unlike the spin-spin coupling between magnetic nuclei which takes place through the
bond of he molecule and this through space effect not lead to coupling. Its magnitude depends on the distance between the
interacting nuclei, even if they are far apart in the bonding network. The NOE effect is generally noticeable only over short
distances, generally 2-5 angstrom. The NOE diminishes dramatically as the distance between saturated and observed nuclei (r)
increases. Its magnitude is inversely proportional to the sixth power of the distance between the nuclei (1 / r 6). These measured
distances are used to determine accurate three-dimensional structures of proteins and nucleic acids.

NOE between nuclei of the same type and of different type are possible. If NOE is carried out on nuclei of a single type,
the process is called the homonuclear Overhauser effect (example between protons). The second and most common example of the
NOE occurs between 1H and 13C in 13C-NMR spectroscopy during the application of proton decoupled spectrum. If NOE is
observed between the different nuclei, it is called the heteronuclear Overhauser effect. The heteronuclear NOE operates in both
direction; either atom can be irradiated.

The degree of increase in the signal is called the nuclear Overhauser enhancement. The maximum enhancement that can be
observed is given by the relationship

where γ irr is the magnetogyric ratio of the nucleus being irradiated, and γ obs is that of the nucleus being observed. NOEmax is
the enhancement of the signal, and it must be added to the original signal strength for the total intensity of a peak.

For a proton-decoupled 13 C spectrum,

Because of the distance dependence of the NOE, it is an important method for providing useful geometrical information
like which groups are close together in space, for determining the three dimensional structure and stereochemistry of organic
compounds.

Because NOE increases the intensity of the carbon signals, it substantially increases the sensitivity (signal-to-noise ratio) in the 13C
spectrum. Because of the enhancement of sensitivity, NOE is generally used for the sensitivity improvement for nuclei having low
magnetogyrric ratio-γ by the magnetization transfer from high-γ nuclei (usually protons) in the same molecule. This is achieved by
cross-relaxation between the two nuclei after the population difference of the two states of the high-γ nucleus is removed from the
equilibrium value, by saturation using a strong radiofrequency.

The intensities of many carbon resonances in a proton-decoupled spectrum increase significantly above those observed in a
proton-coupled spectrum. Carbon atoms with Hs directly attached are more enhanced (more Hs attached, more enhancement) due
to NOE. The intensity of 13C resonance may be increased by up to 200% when 1H nuclei which are directly bonded to C atoms
are irradiated. The effect is very important in increasing the intensity of 13C spectra when they are proton decoupled, because
otherwise the intensity of 13C spectra is low due to low natural abundance of 13C atoms.

In 1H spectra, the maximum enhancement can only be 50% of the usual intensity of the signal, but the usual range is 1-
20%.

The NOE is a powerful method for establishing the three-dimensional proximity of nuclei in space and is used extensively
in mapping and establishing the conformations, configurations and stereochemistry of molecules in solution.

NOE is a useful method for detecting which groups are close in space to each other. The technique provide valuable
structural information, especially for determining stereochemistry.
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Origin of NOE: cross-polarization
Signal enhancement due to NOE is an example of cross-polarization, in which a polarization of the spin states in one type
of nucleus causes a polarization of the spin states in another nucleus.
Irradiation of one nucleus with a weak radio frequency signal at its resonant frequency will equalize the populations in its
two energy levels. This perturbation of population levels disturbs the populations of nearby nuclei so as to enhance the intensity of
absorbance at the resonant frequency of the nearby nuclei.
Irradiation of one nucleus with a weak radio frequency signal at its resonant frequency will equalize the populations in its two
energy levels. This perturbation of population levels disturbs the populations of nearby nuclei so as to enhance the intensity of
absorbance at the resonant frequency of the nearby nuclei. NOE develops during continuous irradiation at even lower power during
the relaxation delay.
The intensity of an NMR signal is directly proportional to the population of the energy levels of the nuclei in a given
magnetic field. The population distribution of a nucleus (difference between populations in the upper spin state and the lower spin
state) can be affected by the population distributions of other nuclei that are nearby in space. According to the Boltzmann
distribution law, there is a slight excess of nuclei aligned with the external magnetic field B0. The intensity of the NMR signals
obtained is directly proportional to the number of the excess nuclei in the lower state. The energy levels of protons and carbon are
established after a thermal equilibrium. If we perturb this thermal equilibrium, for example, by increasing the number of the
protons nuclei populating the higher energy level by irradiating the protons, the system will try to restore the equilibrium and this
will induce a transition so that the population of the lower energy level of 13C will increase. So there will be a corresponding
enhancement in the 13C signal. The process of increasing and decreasing the populations of different energy levels within the
system is called dynamic nuclear polarization.
In proton decoupled 13C NMR, when the hydrogens in the molecule are irradiated, they become saturated and attain a
distribution of spins very different from their equilibrium (Boltzmann) state. There are more spins than normal in the excited state.
Due to the interaction of spin dipoles, the spins of the carbon nuclei “sense” the spin imbalance of the hydrogen nuclei and begin to
adjust themselves to a new equilibrium state that has more spins in the lower state. This increase of population in the lower spin
state of carbon increases the intensity of the NMR signal.