NMR spectroscopy

Quantum numbers and their role in NMR, principle, instrumentation, solvent requirements in NMR, relaxation
process, NMR signals in various compounds, chemical shift, factors influencing chemical shift, spin – spin
coupling, coupling constant, nuclear magnetic double resonance, brief outline and principle of FT-NMR and 13C
NMR. Applications of NMR spectroscopy.
https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Book
%3A_Physical_Methods_in_Chemistry_and_Nano_Science_(Barron)/04%3A_Chemical_Speciation/
4.07%3A_NMR_Spectroscopy
 Gradient Coils
• The gradient coils produce the gradients in the Bo magnetic field needed for performing
gradient enhanced spectroscopy, diffusion measurements, and NMR microscopy. The
gradient coils are located inside the RF probe. Not all probes have gradient coils, and not all
NMR spectrometers have the hardware necessary to drive these coils.
• The gradient coils are room temperature coils (i.e. do not require cooling with cryogens to
operate) which, because of their configuration, create the desired gradient.
• The vertical bore superconducting magnet is most common.
• Assuming the standard magnetic resonance coordinate system, a gradient in Bo in the Z
direction is achieved with an antihelmholtz type of coil. Current in the two coils flow in
opposite directions creating a magnetic field gradient between the two coils. The B field at
the center of one coil adds to the Bo field, while the B field at the center of the other coil
subtracts from the Bo field.
• The X and Y gradients in the Bo field are created by a pair of figure-8 coils. The X axis figure-8
coils create a gradient in Bo in the X direction due to the direction of the current through the
 Quadrature Detector
• The quadrature detector is a device which separates out the Mx' and My' signals from
the signal from the RF coil. For this reason it can be thought of as a laboratory to
rotating frame of reference converter. The heart of a quadrature detector is a device
called a doubly balanced mixer. The doubly balanced mixer has two inputs and one
output. If the input signals are Cos(A) and Cos(B), the output will be 1/2 Cos(A+B) and
1/2 Cos(A-B). For this reason the device is often called a product detector since the
product of Cos(A) and Cos(B) is the output.
• The quadrature detector typically contains two doubly balanced mixers, two filters,
two amplifiers, and a 90o phase shifter. There are two inputs and two outputs on the
device. Frequently ν and ν0 are put in and the MX' and MY' components of the
transverse magnetization come out. There are some potential problems which can
occur with this device which will cause artefacts in the spectrum. One is called a DC
offset artefact and the other is called a quadrature artefact.
 Digital Filtering
• Many newer spectrometers employ a combination of oversampling, digital filtering, and
decimation to eliminate the wrap around artefact. Oversampling creates a larger spectral or
sweep width, but generates too much data to be conveniently stored. Digital filtering
eliminates the high frequency components from the data, and decimation reduces the size
of the data set.
• Oversampling is the digitization of a time domain signal at a frequency much greater than
necessary to record the desired spectral width. For example, if the sampling frequency, f s, is
increased by a factor of 10, the sweep width will be 10 times greater, thus eliminating
wraparound. Unfortunately digitizing at 10 times the speed also increases the amount of raw
data by a factor of 10, thus increasing storage requirements and processing time.
• Filtering is the removal of a select band of frequencies from a signal. In NMR, this step would
be equivalent to taking a large sweep width spectrum and setting to zero intensity those
spectral frequencies which are farther than some distance from the center of the spectrum.
• Digital filtering is the removal of these frequencies using the time domain signal. We know that if
two functions are multiplied in one domain (i.e. frequency), we must convolve the FT of the two
functions together in the other domain (i.e. time). To filter out frequencies above fo from the time
domain signal, the signal must be convolved with the Fourier transform of the rectangular
function, a sin c function. This process eliminates frequencies greater than fo from the time
domain signal. Fourier transforming the resultant time domain signal yields a frequency domain
signal without the higher frequencies. In NMR, this step will remove spectral components with
frequencies greater than +fo and less than -fo.
• Decimation is the elimination of data points from a data set. A decimation ratio of 4/5 means that
4 out of every 5 data points are deleted, or every fifth data point is saved. Decimating the digitally
filtered data, followed by a Fourier transform, will reduce the data set by a factor of five.
• High speed digitizers, capable of digitizing at 2 MHz, and dedicated high speed integrated circuits,
capable of performing the convolution on the time domain data as it is being recorded, are used
to realize this procedure.
 Safety
• There are some important safety considerations which one should be familiar with before
using an NMR spectrometer.
• These concern the use of strong magnetic fields and cryogenic liquids.
• Magnetic fields from high field magnets can literally pick up and pull ferromagnetic items
into the bore of the magnet.
• Keep all ferromagnetic items away from the magnet because they can seriously damage the magnet,
shim coils, and probe.
• The force exerted on the concentric cryogenic dewars within a magnet by a large metal object stuck
to the magnet can break dewars and magnet supports.
• The kinetic energy of an object being sucked into a magnet can smash a dewar or an electrical
connector on a probe.
• Small ferromagnetic objects are just as much a concern as larger ones.
• A small metal sliver can get sucked into the bore of the magnet and destroy the homogeneity of the
magnet achieved with a set of shim settings.
• There are additional concerns regarding the effect of magnetic fields on electronic circuitry,
specifically pacemakers.
• An individual with a pacemaker walking through a strong magnetic field can induce currents in the
pacemaker circuitry which will cause it to fail and possibly cause death.
• A person with a pacemaker should never venture near NMR spectrometer.
• Mechanical watches and some digital watches will also be affected by magnetic fields.
• Magnetic fields of approximately 50 Gauss will erase credit cards and magnetic storage media.
• The liquid nitrogen and liquid helium used in NMR spectrometers are at a temperature of 77.4 K
and 4.2 K respectively.
• These liquids can cause frostbite, which is a concern while filling the magnet.
• Suffocation is another concern when filling the magnet. If the magnet quenches, or suddenly stops
being a superconductor, it will rapidly boil off all its cryogens, and the nitrogen and helium gasses
in a confined space can cause suffocation.
 Solvent requirements in NMR
• Most NMR spectra are recorded for compounds dissolved in a solvent.
• Therefore, signals will be observed for the solvent and this must be accounted for
in solving spectral problems.
• NMR solvents are distinctly different from other spectroscopic solvents as majority
of hydrogen nuclei are replaced with deuterium so as to minimize the interference
due to protons. However, deuteration is not "100%", so signals for the residual
protons are observed.
• It used to be common practice to add (0.03%) tetra methyl silane, or related
compounds, as an internal reference standard for 1H and 13C NMR spectra with the
proton signal occurring at 0.0 ppm and the carbon signal occurring at 0.0 ppm in
the 13C NMR spectrum. However, modern spectrometers can "lock" on solvent
signals, so addition of internal reference standards is not usually required.
 The chemical shifts (d) of solvent signals
observed with multiplicity in paranthesis
Solvent H NMR Chemical Shift
1 13
C NMR Chemical Shift

Acetic Acid 11.65 (1) , 2.04 (5) 179.0 (1) , 20.0 (7)

Acetone 2.05 (5) 206.7 (13) , 29.9 (7)

Acetonitrile 1.94 (5) 118.7 (1) , 1.39 (7)

Benzene 7.16 (1) 128.4 (3)

Chloroform 7.26 (1) 77.2 (3)

Dimethyl Sulfoxide 2.50 (5) 39.5 (7)

Methanol 4.87 (1) , 3.31 (5) 49.1 (7)

Methylene Chloride 5.32 (3) 54.00 (5)

Pyridine 8.74 (1) , 7.58 (1) , 7.22 (1) 150.3 (1) , 135.9 (3) , 123.9 (5)

Water (D2O) 4.8

 Desirable characteristics of NMR solvents
are:
• Solubility
Higher solubility of sample in the chosen solvent contributes to higher sensitivity. This property assumes significance in cases where
sample availability is limited.
• Purity
Degree of solvent purity is a major selection criteria. Chemical impurities or water content can lead to interference signals in the NMR
spectrum.
High purity solvents are available in sealed glass bottles. After use the bottle should be immediately closed to prevent intake of
moisture or other impurities from the atmosphere.
Solvent blanks should be run in advance so as to determine that the solvent peaks do not interfere with the sample peaks.
• Solvent viscosity
Lower the sample viscosity the better will be the spectral resolution due to better homogenization of the sample.
• Moisture Content
Presence of traces of water will contribute to spectral interferences. Moisture content can be reduced by filtration of solvent through a
drying agent or storing the solvent over molecular sieves.
• Price
The solvent price is of concern especially when large number of samples need to be analysed. The price goes up further as the degree
of deuteration increases.
Commonly used NMR solvents
• Deuterated water-D2O
• Deuterated chloroform,CDCl3
• DMSO−d6
• Methanol−d4
• Methylenechloride−d2
• Pyridine−d5
• THF−d8
• Aceticacid−d4
• Acetone−d6
• CD3CN
Deuterated chloroform,CDCl3 is most commonly used because of its low price.
Chemical Shift and
Factors influencing
the same
The frequency at which a particular
proton comes into resonance is known
as its chemical shift.
1
H-NMR plot for methyl
acetate.
Just as in IR and UV-vis spectroscopy, the
vertical axis corresponds to intensity of
absorbance, the horizontal axis to frequency
(typically the vertical axis is not shown in an
NMR spectrum).
We see three absorbance signals: two of
these correspond to Ha and Hb, while the
peak at the far right of the spectrum
corresponds to the 12 chemically equivalent
protons in tetramethylsilane (TMS), a
standard reference compound that was
added to the sample.
• The NMR spectra is displayed as a plot of the applied radio frequency versus the absorption.
• The position on the plot at which the nuclei absorbs is called the chemical shift. Since this has an
arbitrary value a standard reference point must be used. The two most common standards are
TMS (tetramethylsilane, (Si(CH3)4) which has been assigned a chemical shift of zero, and
CDCl3 (deuterochloroform) which has a chemical shift of 7.26 for 1H NMR and 77 for 13C NMR.
• The scale is commonly expressed as parts per million (ppm) which is independent of the
spectrometer frequency. The scale is the delta (δ) scale.

• The range at which most NMR absorptions occur is quite narrow. Almost all 1H absorptions occur
downfield within 10 ppm of TMS. For 13C NMR almost all absorptions occurs within 220 ppm
downfield of the C atom in TMS.
 Shielding in NMR
• Structural features of the molecule will have an effect on the exact magnitude of the
magnetic field experienced by a particular nucleus. This means that H atoms which
have different chemical environments will have different chemical shifts. This is what
makes NMR so useful for structure determination in organic chemistry. There are
three main features that will affect the shielding of the nucleus, electronegativity,
magnetic anisotropy of π systems and hydrogen bonding.
• Electronegativity: The electrons that surround the nucleus are in motion so they
created their own electromagnetic field. This field opposes the applied magnetic field
and so reduces the field experienced by the nucleus. Thus the electrons are said
to shield the nucleus. Since the magnetic field experienced at the nucleus defines the
energy difference between spin states it also defines what the chemical shift will be
for that nucleus. Electron with-drawing groups can decrease the electron density at
the nucleus, deshielding the nucleus and result in a larger chemical shift.
• Magnetic Anisotropy: π Electron
Effects: The π electrons in a
compound, when placed in a
magnetic field, will move and
generate their own magnetic
field. The new magnetic field will
have an effect on the shielding
of atoms within the field.
• Hydrogen Bonding
• Protons that are involved in hydrogen
bonding (i.e.-OH or -NH) are usually
observed over a wide range of chemical
shifts. This is due to the deshielding that
occurs in the hydrogen bond. Since
hydrogen bonds are dynamic, constantly
forming, breaking and forming again, there
will be a wide range of hydrogen bonds
strengths and consequently a wide range of
deshielding. This as well as solvation
effects, acidity, concentration and
temperature make it very difficult to predict
the chemical shifts for these atoms.
 Diamagnetic shielding and
deshielding effects
• The chemical shift of a given proton is determined primarily by its immediate electronic environment.
• In methane molecule (CH4), valence electrons around the methyl carbon, when subjected to B0, are induced to circulate
and thus generate their own very small magnetic field that opposes B0.
• This induced field, to a small but significant degree, shields the nearby protons from experiencing the full force of B0, an
effect known as local diamagnetic shielding.
• The methane protons therefore do not experience the full force of B0 - what they experience is called Beff, or the effective
field, which is slightly weaker than B0.
• Methyl fluoride, CH3F, has higher chemical shift than methane due to deshielding effect.
• As fluorine is more electronegative than carbon, it pulls valence electrons away from the carbon,
effectively decreasing the electron density around each of the protons. For the protons, lower electron density means less
diamagnetic shielding, which in turn means a greater overall exposure to B0, a stronger Beff, and a higher resonance
frequency. Put another way, the fluorine, by pulling electron density away from the protons, is deshielding them, leaving
them more exposed to B0.
• As the electronegativity of the substituent increases, so does the extent of deshielding, and so does the chemical shift.
This is evident when we look at the chemical shifts of methane and three halomethane compounds (remember that
electronegativity increases as we move up a column in the periodic table).
• Chemical shift can be predicted by
considering deshielding near a proton. The
chemical shift of trichloromethane is,
higher than that of dichloromethane,
which is in turn higher than that of
chloromethane.
• The deshielding effect of an
electronegative substituent diminishes
sharply with increasing distance.
• The presence of an electronegative oxygen,
nitrogen, sulphur, or sp2-hybridized carbon
also tends to shift the NMR signals of
nearby protons slightly downfield.
• The signal at 3.65 ppm in methyl acetate
corresponds to the methyl ester protons (Hb),
which are deshielded by the adjacent oxygen
atom. The upfield signal at 2.05 ppm
corresponds to the acetate protons (Ha), which
is deshielded - but to a lesser extent - by the
adjacent carbonyl group.
• One of the main reasons why the TMS proton
signal was chosen as a zero-point is that the
TMS protons are highly shielded: silicon is
slightly less electronegative than carbon, and
therefore donates some additional shielding
electron density. Very few organic molecules
contain protons with chemical shifts that are
negative relative to TMS.
 Factors affecting chemical shift
• Hydrogen Bonding:
• In general hydrogen bonded protons show higher value of chemical shift than the non-hydrogen bonded
protons.
• Due to the high electro-negativity of the atom to which proton is hydrogen bonded, the electron cloud
around the proton is decreased and thus causes deshielding of the proton.
• The downfield shift of the absorption depends on the strength of the hydrogen bonding. Greater is the
degree of hydrogen bonding of proton, greater the downfield shift of a proton.
• This is due to hydrogen bonding that the chemical shifts of OH, NH and SH protons vary over a wide range.
• It is unpredictable as the extent of hydrogen bonding depends on concentration, temperature, solvent and
compound structure.
• The OH signals tend to move downfield at higher compound concentration because of increased hydrogen
bonding.
• Also on increasing temperature, hydrogen bonding decreases and hence OH signals moves slightly up field.
• Both OH and NH signals move downfield in H-bonding solvents like DMSO or acetone.
• The intermolecular and intramolecular hydrogen
bonding can be distinguished by the use of 1H
NMR spectroscopy. The intermolecular hydrogen
bonding depends on the concentration of the
sample and decreases on dilution with a non-
polar solvent and also with increasing
temperature. The pure ethanol show signals near
5 ppm but on dilution with non hydrogen-bonded
solvents (CCl4, CDCl3, C6D6), the OH signal shift
up field due to the breaking of intermolecular
hydrogen bonds. The more acidic hydroxyl group
of phenol also shows similar concentration
dependence to that of alcohols. The phenolic OH
peak at different percent concentrations of
phenol in CDCl3 are shown in the following
diagram (C-H signals are not shown).
• The intramolecular hydrogen bonding does not depend on the
concentration of the sample and hence show no change in their
absorption position on dilution. The salicylates, enols of β-diketones
and carboxylic acids are some classic examples of intramolecular
hydrogen bonding. The hydroxyl proton of enol form of 2,4-
pentanedione (4-hydroxypent-3-ene-2-one) shows a peak significantly
down field (10-16 ppm) compared to the OH signal in other function
groups due to strong intramolecular hydrogen bonding. Similarly the
hydroxyl proton of carboxylic acid displays a very downfield peak at
about 10-13 ppm which may be attributed to the favored hydrogen-
bonded dimeric association.
 Solvent Effects
• Chloroform (CDCl3) is the most common solvent for NMR measurements.
• Other deuterium labelled compounds, such as deuterium oxide (D2O), benzene-d6 (C6D6), acetone-d6
(CD3COCD3) and DMSO-d6 (CD3SOCD3) are also used as NMR solvents.
• All these solvents vary considerably in their polarity and magnetic susceptibility. Hence the NMR
spectrum recorded in one solvent may be slightly different from that recorded in other solvent.
• Hence it is important to mention the solvent used.
• The NMR signals for protons attached to carbon are generally shifted slightly by changing solvent except
where significant bonding or dipole-dipole interaction might arise.
• The changes in chemical shifts due to solvents changes are minor being on the order of ±0.1 ppm.
• However, in some cases the changes are more. The aromatic solvents benzene and pyridine cause
changes in chemical shifts as large as 0.5 to 0.8 ppm compared to less magnetically active solvents like
chloroform or acetone.
• It is observed that the carbonyl groups form weak π–π collision complexes with benzene rings that
persist long enough to exert a significant shielding influence on nearby groups.
 Shielding and deshielding in Aromatic
Protons
• Electron donating substituents cause shielding of the aromatic
protons as they increase electron density on the benzene ring.
• The order of shielding is ortho > para > meta because the electron
density at ortho and para positions are the higher due to the +M
effect.
• On the other hand, the electron withdrawing substituents cause
deshielding of all the aromatic protons.
• The order of deshielding is in the order of ortho > para > meta
because the electron withdrawing effect is maximum at ortho and
para positions due to the +M effect.
 Anisotropic effect (space effect)
• Anisotropy means uneven magnetic field in space
i.e. different regions of space are characterized by
different magnetic field strengths. The anisotropic
effect depends upon the orientation of the molecule
with respect to the applied field.
• In fact, a proton experiences three different fields,
the applied field, the induced magnetic field
generated by σ-electrons and the induced magnetic
field generated by π-electrons. The π- electrons are
more polarizable than the sigma bond electrons.
Therefore, the induced magnetic field due to π-
electrons movement is stronger (strong secondary
fields that perturb nearby nuclei) than that
generated by σ-electrons. Depending on the
position of the proton in the induced magnetic field,
it can be either shielded (smaller δ) or deshielded
(larger δ).
 Coupling Constant
• NMR spectra of most organic molecules contain proton signals that
are 'split' into two or more sub-peaks.
• Rather than being a complication, however, this splitting behavior is
actually very useful because it provides us with more information
about our sample molecule.
• The terms 'splitting' and 'coupling' are often used interchangeably
when discussing NMR.
• The source of signal splitting is a phenomenon called spin-spin
coupling, a term that describes the magnetic interactions between
neighbouring, non-equivalent NMR-active nuclei.
 Coupling Constant
• The signal at 3.96 ppm, corresponding
to the two Ha protons, is split into two
subpeaks of equal height (and area) –
this is referred to as a doublet. (1:1)
• The Hb signal at 5.76 ppm, on the other
hand, is split into three sub-peaks, with
the middle peak higher than the two
outside peaks - if we were to integrate
each subpeak, we would see that the
area under the middle peak is twice
that of each of the outside peaks. This is
called a triplet. (1:2:1)

1, 1, 2 trichloroethane
• The source of signal splitting is a phenomenon called spin-spin coupling, a term that describes the magnetic interactions between
neighboring, non-equivalent NMR-active nuclei.
• The Ha and Hb protons are spin-coupled to each other.
• Ha proton is also influenced by the small magnetic field generated by Hb proton and shielded by nearby valence electrons.
• The magnetic moment of Hb will be aligned with B0 in slightly more than half of the molecules in the sample, while in the remaining
molecules it will be opposed to B0.
• The Beff ‘felt’ by Ha is a slightly weaker if Hb is aligned against B0, or slightly stronger if Hb is aligned with B0.
• In other words, in half of the molecules Ha is shielded by Hb (thus the NMR signal is shifted slightly upfield) and in the other half Ha is
deshielded by Hb (and the NMR signal shifted slightly downfield).
• What would otherwise be a single Ha peak has been split into two sub-peaks (a doublet), one upfield and one downfield of the
original signal.
• The magnetic environment experienced by Hb is influenced by the fields of both neighboring Ha protons, which we will call Ha1 and
Ha2.
• There are four possibilities here, each of which is equally probable.
• First, the magnetic fields of both Ha1 and Ha2 could be aligned with B0, which would deshield Hb, shifting its NMR signal
slightly downfield.
• Second, both the Ha1 and Ha2 magnetic fields could be aligned opposed to B0, which would shield Hb, shifting its resonance
signal slightly upfield.
• Third and fourth, Ha1 could be with B0 and Ha2 opposed, or Ha1 opposed to B0 and Ha2 with B0.
• In each of the last two cases, the shielding effect of one Ha proton would cancel the deshielding effect of the other, and the chemical
shift of Hb would be unchanged.
• So in the end, the signal for Hb is a triplet, with the middle peak twice as large as the two outer peaks because there are two ways
that Ha1 and Ha2 can cancel each other out.
• Ethyl Acetate
• The splitting of NMR signal gives precise
information about the number of
neighbouring protons in a molecule.
There is a formula to calculate the
multiplicity of the peaks in the NMR
spectrum.
2nI + 1
n= Number of neighbouring protons
I= spin number of protons
• Since I is always ½, we can rewrite the
formula as n+1.
• Pascal’s triangle gives the area ratio of the
peaks.
 Coupling constant and factors affecting
• The separation distance between two adjacent peaks, as a result of the spin-spin interaction in a multiplet, is constant and is known as
coupling constant (denoted by the letter J).
• The unit of a coupling constant is Hz and it is also referred as Cycles per second (CPS).
• The value of the coupling constant could be either positive or negative.
• When two spinning nuclei are in the opposite orientation then the energy is lower and the value of the constant is positive.
• However, if the spinning nuclei are in the same orientation, then, the energy is higher and the value of the constant is negative.
• The spacing between the split lines or the J constant between coupling protons is of the same magnitude.
• The J constant can be used for distinguishing, e.g., between two singlets and one doublet or two doublet and one quartet. There is no
effect of the external magnetic field on the coupling constant.
• The value of the coupling constant depends on the following factors:
Distance between the protons
• The distance between the hydrogen atoms in a molecule is an important determinant in the value of J constant. If the hydrogen atoms
involved in the coupling are closer to each other, these give rise to a greater value of J constant than if these atoms are further apart.
The orientation of the coupled protons
• The orientation or angle of the protons with respect to each other is equally important. The value of J constant is greater in molecules,
where the H atoms are in the cis conformation. Conversely, it is less when the H atoms are in the trans conformation.
 Types of couplings and their effects on the
coupling constant
• Geminal means that 2 atoms or functional groups are bound to the
same carbon atom. The geminal or 2-bond coupling constant is
denoted by 2J.
• The protons i and ii are geminal protons however, their electronic
environments are different.
• The coupling constant value will also depend on the angle between
protons i and ii. The coupling constant will increase with the
electronegativity of the groups.
• Vicinal Coupling:  Vicinal protons are those which are separated by
three bonds. In the molecule shown above, ii and iii are vicinal
protons. The vicinal or 3 bond coupling constants is denoted by 3J.
• Long range coupling:  If the distance between two protons is more
than 3 covalent bonds, then, the phenomenon of coupling does not
come into play. However, there would be some coupling if there are
unsaturated or fluoro compounds present in the vicinity of the
protons. Such type of coupling can only be observed using a very
sensitive and high-resolution NMR spectrophotometer.
• Complex Splitting Patterns: The NMR spectrum can sometimes
have more complex splitting patterns & coupling constants like
doublet of doublets, doublet of tripltes etc.