Nuclear Magnetic Resonance Spectros

H1-Nuclear Magnetic Resonance Spectroscopy MPY-102
1. INTRODUCTION
Over the past fifty years nuclear magnetic resonance spectroscopy, commonly
referred to as 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. Although larger amounts of sample are needed than for mass spectroscopy,
NMR is non-destructive, and with modern instruments good data may be obtained
from samples weighing less than a milligram. To be successful in using NMR as an
analytical tool, it is necessary to understand the physical principles on which the
methods are based.
The nuclei of many elemental isotopes have a characteristic spin (I). Some nuclei
have integral spins (e.g. I = 1, 2, 3 ....), some have fractional spins (e.g. I = 1/2, 3/2,
5/2 ....), and a few have no spin, I = 0 (e.g. 12C, 16O, 32S ...). Isotopes of particular
interest and use to organic chemists are 1H, 13C, 19F and 31P, all of which have I = 1/2.
Since the analysis of this spin state is fairly straightforward, our discussion of NMR
will be limited to these and other I = 1/2 nuclei.
2. SPIN PROPERTIES OF NUCLEI
Nuclear spin may be related to the nucleon composition of a nucleus in the following
manner:
• Odd mass nuclei (i.e. those having an odd number of nucleons) have fractional
spins. Examples are I = 1/2 (1H, 13C, 19F), I = 3/2 (11B) & I = 5/2 (17O).
• Even mass nuclei composed of odd numbers of protons and neutrons have
integral spins. Examples are I = 1 (2H, 14N).
• Even mass nuclei composed of even numbers of protons and neutrons have
zero spin ( I = 0 ). Examples are 12C, and 16O.
Spin 1/2 nuclei have a spherical charge distribution, and their NMR behavior is the
easiest to understand. Other spin nuclei have non-spherical charge distributions and
may be analyzed as prolate or oblate spinning bodies. All nuclei with non-zero spins
School of Pharmacy, SRTMU Nanded Page 1 of 41

have magnetic moments (μ), but the non-spherical nuclei also have an electric
quadrupole moment (eQ). Some characteristic properties of selected nuclei are given
in the following table.
Natural % Magnetic Magnetogyric

Isotope Spin (I)
Abundance Moment (μ)* Ratio (γ)†
1
H 99.9844 1/2 2.7927 26.753
2
H 0.0156 1 0.8574 4,107
11
B 81.17 3/2 2.6880 --
13
C 1.108 1/2 0.7022 6,728
17
O 0.037 5/2 -1.8930 -3,628
19
F 100.0 1/2 2.6273 25,179
29
Si 4.700 1/2 -0.5555 -5,319
31
P 100.0 1/2 1.1305 10,840
* μ in units of nuclear magnetons = 5.05078•10-27 JT-1
†
γ in units of 107rad T-1 sec-1
3. FEATURES THAT LEAD TO THE NMR PHENOMENON:
1. A spinning charge generates a magnetic field, as shown by the animation on the

right.
The resulting spin-magnet has a magnetic moment (μ) proportional to the spin.
2. In the presence of an external magnetic field (B0), two spin states exist, +1/2 and
-1/2. The magnetic moment of the lower energy +1/2 state is alligned with the
external field, but that of the higher energy -1/2 spin state is opposed to the external
field. Note that the arrow representing the external field points north.
3. The difference in energy between the two spin states is dependent on the external
magnetic field strength, and is always very small. The following diagram illustrates
that the two spin states have the same energy when the external field is zero, but
diverge as the field increases. At a field equal to Bx a formula for the energy
difference is given (remember I = 1/2 and μ is the magnetic moment of the nucleus
in the field).

Strong magnetic fields are necessary for NMR spectroscopy. The international unit
for magnetic flux is the tesla (T). The earth's magnetic field is not constant, but is
approximately 10-4 T at ground level. Modern NMR spectrometers use powerful
magnets having fields of 1 to 20 T. Even with these high fields, the energy
difference between the two spin states is less than 0.1 cal/mole. To put this in
perspective, recall that infrared transitions involve 1 to 10 kcal/mole and electronic
transitions are nearly 100 times greater.
For NMR purposes, this small energy difference (ΔE) is usually given as a
frequency in units of MHz (106 Hz), ranging from 20 to 900 Mz, depending on the
magnetic field strength and the specific nucleus being studied. Irradiation of a
sample with radio frequency (rf) energy corresponding exactly to the spin state
separation of a specific set of nuclei will cause excitation of those nuclei in the +1/2
state to the higher -1/2 spin state. Note that this electromagnetic radiation falls in
the radio and television broadcast spectrum. NMR spectroscopy is therefore the
energetically mildest probe used to examine the structure of molecules.
The nucleus of a hydrogen atom (the proton) has a magnetic moment μ = 2.7927,
and has been studied more than any other nucleus.
4. For spin 1/2 nuclei the energy difference between the two spin states at a given
magnetic field strength will be proportional to their magnetic moments. For the four
common nuclei noted above, the magnetic moments are: 1H μ = 2.7927, 19F μ =
2.6273, 31P μ = 1.1305 & 13C μ = 0.7022. These moments are in nuclear magnetons,
which are 5.05078•10-27 JT-1. The following diagram gives the approximate
frequencies that correspond to the spin state energy separations for each of these
nuclei in an external magnetic field of 2.35 T. The formula in the colored box shows

the direct correlation of frequency (energy difference) with magnetic moment (h =

Planck's constant = 6.626069•10-34 Js).
4. NUCLEAR SPIN AND THE SPLITTING OF ENERGY LEVELS

IN A MAGNETIC FIELD
Subatomic particles (electrons, protons and neutrons) can be imagined as spinning on

their axes. In many atoms (such as 12C) these spins are paired against each other, such
that the nucleus of the atom has no overall spin. However, in some atoms (such as 1H
and 13C) the nucleus does possess an overall spin. The rules for determining the net
spin of a nucleus are as follows;
1. If the number of neutrons and the number of protons are both even, then the
nucleus has NO spin.
2. If the number of neutrons plus the number of protons is odd, then the nucleus
has a half-integer spin (i.e. 1/2, 3/2, 5/2)
3. If the number of neutrons and the number of protons are both odd, then the
nucleus has an integer spin (i.e. 1, 2, 3)
The overall spin, I, is important. Quantum mechanics tells us that a nucleus of spin I
will have 2I + 1 possible orientations. A nucleus with spin 1/2 will have 2 possible
orientations. In the absence of an external magnetic field, these orientations are of
equal energy. If a magnetic field is applied, then the energy levels split. Each level is
given a magnetic quantum number, m.

When the nucleus is in a magnetic field, the initial populations of the energy levels are
determined by thermodynamics, as described by the Boltzmann distribution. This is
very important, and it means that the lower energy level will contain slightly more
nuclei than the higher level. It is possible to excite these nuclei into the higher level
with electromagnetic radiation. The frequency of radiation needed is determined by
the difference in energy between the energy levels.
5. CALCULATING TRANSITION ENERGY
The nucleus has a positive charge and is spinning. This generates a small magnetic
field. The nucleus therefore possesses a magnetic moment, , which is proportional
to its spin, I.
The constant, γ, is called the magnetogyric ratio and is a fundamental nuclear

constant which has a different value for every nucleus. h is Planck’s constant.
The energy of a particular energy level is given by;
Where B is the strength of the magnetic field at the nucleus.
The difference in energy between levels (the transition energy) can be found from

This means that if the magnetic field, B, is increased, so is ΔE. It also means that if a
nucleus has a relatively large magnetogyric ratio, then ΔE is correspondingly large.
If you had trouble understanding this section, try reading the next bit (The absorption
of radiation by a nucleus in a magnetic field) and then come back.
6. RELAXATION PROCESSES
How do nuclei in the higher energy state return to the lower state? Emission of
radiation is insignificant because the probability of re-emission of photons varies with
the cube of the frequency. At radio frequencies, re-emission is negligible. We must
focus on non-radiational relaxation processes (thermodynamics!).
Ideally, the NMR spectroscopist would like relaxation rates to be fast - but not too
fast. If the relaxation rate is fast, then saturation is reduced. If the relaxation rate is too
fast, line-broadening in the resultant NMR spectrum is observed.
There are two major relaxation processes;
• Spin - lattice (longitudinal) relaxation

• Spin - spin (transverse) relaxation
6.1 Spin - lattice relaxation
Nuclei in an NMR experiment are in a sample. The sample in which the nuclei are
held is called the lattice. Nuclei in the lattice are in vibrational and rotational motion,
which creates a complex magnetic field. The magnetic field caused by motion of
nuclei within the lattice is called the lattice field. This lattice field has many
components. Some of these components will be equal in frequency and phase to the
Larmor frequency of the nuclei of interest. These components of the lattice field can
interact with nuclei in the higher energy state, and cause them to lose energy
(returning to the lower state). The energy that a nucleus loses increases the amount of

vibration and rotation within the lattice (resulting in a tiny rise in the temperature of
the sample).
The relaxation time, T1 (the average lifetime of nuclei in the higher energy state) is
dependant on the magnetogyric ratio of the nucleus and the mobility of the lattice. As
mobility increases, the vibrational and rotational frequencies increase, making it more
likely for a component of the lattice field to be able to interact with excited nuclei.
However, at extremely high mobilities, the probability of a component of the lattice
field being able to interact with excited nuclei decreases.
6.2 Spin - spin relaxation
Spin - spin relaxation describes the interaction between neighbouring nuclei with
identical precessional frequencies but differing magnetic quantum states. In this
situation, the nuclei can exchange quantum states; a nucleus in the lower energy level
will be excited, while the excited nucleus relaxes to the lower energy state. There is
no net change in the populations of the energy states, but the average lifetime of a
nucleus in the excited state will decrease. This can result in line-broadening.

7. NUMBER OF SIGNALS
7.1 Protons with identical chemical environments:
A collection of identical nuclei will all precess with the Larmor frequency but with
random phases relative to each other such that on average the overall (macroscopic)
magnetisation vector Mo remains along the z axis (a).
Imagine now some electromagnetic energy (in the radio-frequency region of the
spectrum) produced by an oscillator (transmitter) and continuously applied to this
system in such a way that its magnetic component B1 is orientated along the x axis, ie
perpendicular to the applied magnetic field. If this electromagnetic field has a specific
oscillation frequency, this can be resolved into two rotational components +w and -w
with respect to the z axis. If we adjust things such that +w of the applied field matches
the Larmor frequency wo exactly, it is said to be in resonance with the precession of
our group of protons. Two things can be thought of as happening when this resonance
occurs;
1. The resonance provides the energy to "flip" some of the proton nuclei from the
ground to the excited state
2. The energy can only be transferred to any individual precessing proton if its
relative phase is the same as that of the precession of the magnetisation
vectors becomes coherent.

If the phases of a proportion of the precessing protons are coherent, the individual
magnetisation vectors no longer cancel each other (b above). Instead the overall
macroscopic vector "tips" away from Mo to a new value M by some angle q with
respect to the z axis. Because the population in the ground (spin +1/2) state is very
slightly greater than the opposing state, M is shown as a positive component in the z
axis in the diagram above. The new magnetisation vector M now precesses about
about the z axis and produces a new non-zero magnetisation component in the xy
plane which did not exist before B1 was applied. Note that if M tips away from Mo
by an angle q of 90, the populations of the ground (M) and the excited (-M) states
become equalised. If the angle q is 180, the populations effectively invert, ie there is a
slight preponderance of the excited state. At this point, it is not possible for the nuclei
to absorb any more energy. There are however several mechanisms whereby the
nuclei can loose energy, or relax. The first involves transfer of energy from the
nucleus to surrounding molecules as thermal motion (heat) by a process known as
spin-lattice or longitudinal relaxation (see below) and the second involves energy
transfer to nuclei of a different type (ie with a different chemical shift) referred to as
spin-spin or transverse relaxation. The rate of relaxation is exponential and is defined
by time constants called T1 and T2 respectively (for protons these have values of ca
0.2 - 5 seconds).
This relaxation phenomenon will have implications for the "width" of the subsequent
spectral lines we will measure (see later).
We are now in a position to measure experimentally the frequency of the process

leading to the non-zero magnetisation component in the xy plane which did not exist
before B1 was applied. This is done by placing a coil (receiver) along the y axis of
our coordinate system and measuring the y component magnetisation of the
precessing vector M. When resonance occurs, the oscillating current induced in the
receiver is detected, amplified and displayed as an "NMR signal". Note particularly

that we measure only the xy component magnetisation of M, and NOT absorption or

emission of energy by the protons. [Actually, we could measure the excited state
relaxation by microcalorimetry, ie the solution warms up slightly when resonance
occurs!].
7.2. Protons with different electronic environments.

The "diamagnetic shielding" of the applied magnetic field by the electronic
environment of each different nucleus in our collection produces slightly different
local values of Bo and hence different characteristic Larmor frequencies for each
unique nucleus.
Beff = (1-s) Bo
Where, s describes the shielding of the effective magnetic field by the electrons
surrounding the nucleus. The experiment described previously can be modified to
detect nuclei resonating at different frequencies by making the transmitter sweep
through the range of expected frequencies and recording the receiver response for
each resonance. A machine built to such a specification is called a continuous wave or
CW machine, such as one of the R24 machines in the teaching laboratories.
(Technically, it is actually easier to adjust Bo than +w on some machines, but that is a
minor detail). However, a CW machine has one major disadvantage, which is that
only one resonant frequency can be measured at any instant. For protons, the detected
frequencies can differ by 600 Hz or more (at 60 MHz) and the time constants for
relaxation described above limit the detectable response rate of the protons. In effect,
this means that one cannot sweep through this frequency range at a rate of more than
about 1Hz per second, i.e. it would take 600 seconds to record the spectrum.

8. INSTRUMENTATION
8.1 Overview
Diagram below displays a schematic representation of the major systems of a nuclear
magnetic resonance spectrometer and a few of the major interconnections. This
overview briefly states the function of each component. Some will be described in
detail later.
At the top of the schematic representation, you will find the superconducting magnet
of the NMR spectrometer. The magnet produces the Bo field necessary for the NMR
experiments. Immediately within the bore of the magnet are the shim coils for
homogenizing the Bo field. Within the shim coils is the probe. The probe contains the
RF coils for producing the B1 magnetic field necessary to rotate the spins by 90o or
180o. The RF coil also detects the signal from the spins within the sample. The sample
is positioned within the RF coil of the probe. Some probes also contain a set of
gradient coils. These coils produce a gradient in Bo along the X, Y, or Z axis. Gradient
coils are used for gradient enhanced spectroscopy, diffusion, and NMR microscopy
experiments.

The heart of the spectrometer is the computer. It controls all of the components of the
spectrometer. The RF components under control of the computer are the RF
frequency source and pulse programmer. The source produces a sine wave of the
desired frequency. The pulse programmer sets the width, and in some cases the shape,
of the RF pulses. The RF amplifier increases the pulses power from milli Watts to
tens or hundreds of Watts. The computer also controls the gradient pulse programmer
which sets the shape and amplitude of gradient fields. The gradient amplifier
increases the power of the gradient pulses to a level sufficient to drive the gradient
coils.
The operator of the spectrometer gives input to the computer through a console
terminal with a mouse and keyboard. Some spectrometers also have a separate small
interface for carrying out some of the more routine procedures on the spectrometer. A
pulse sequence is selected and customized from the console terminal. The operator
can see spectra on a video display located on the console and can make hard copies of
spectra using a printer.
The next sections of this chapter go into more detail concerning the magnet, lock,
shim coils, gradient coils, RF coils, and RF detector of nuclear magnetic resonance
spectrometer.
8.2 Magnet
In most current NMR spectrometers the magnetic field is generated by a
superconducting magnet (Figure below). The first stage in reaching the very low
temperature needed is an outer stainless steel or aluminum dewar which contains
liquid nitrogen. Typically, this has to be refilled every ten days. In practice, it is
advisable to do this refilling on a fixed day every week. An inner dewar contains the
superconducting coil (4) immersed in liquid helium, which has to be refilled,
depending on the construction, every two to eight months. The helium refill should be
carried out only by experienced people. A room-temperature bore is fitted with the
shim coils (7), providing a room-temperature homogeneity adjustment, and a spinner
assembly (5), which contains the turbine system for spinning the NMR sample tube.

The probe-head (8) is usually introduced into the magnet from the bottom and is
connected to at least three radiofrequency (r.f.) cables providing the 2H lock, 1H
frequency, and one X-nucleus frequency. Additional devices to control temperature
(heater, thermoelement, air, sometimes water to insulate the probe-head from the
magnet) are needed. New developments include the digital transmission of the probe-
head parameters to the console via a data line.
8.3 Field Lock

In order to produce a high resolution NMR spectrum of a sample, especially one
which requires signal averaging or phase cycling, you need to have a temporally
constant and spatially homogeneous magnetic field. Consistency of the Bo field over
time will be discussed here; homogeneity will be discussed in the next section of this
chapter. The field strength might vary over time due to aging of the magnet,
movement of metal objects near the magnet, and temperature fluctuations. Here is an
example of a one line NMR spectrum of cyclohexane recorded while the B o magnetic
field was drifting a very significant amount. The field lock can compensate for these
variations.

The field lock is a separate NMR spectrometer within your spectrometer. This
spectrometer is typically tuned to the deuterium NMR resonance frequency. It
constantly monitors the resonance frequency of the deuterium signal and makes minor
changes in the Bo magnetic field to keep the resonance frequency constant. The
deuterium signal comes from the deuterium solvent used to prepare the sample. The
figure below contains plots of the deuterium resonance lock frequency, the small
additional magnetic field used to correct the lock frequency, and the resultant Bo field
as a function of time while the magnetic field is drifting. The lock frequency plot
displays the frequency without correction. In reality, this frequency would be kept
constant by the application of the lock field which offsets the drift.
On most NMR spectrometers the deuterium lock serves a second function. It provides
the =0 reference. The resonance frequency of the deuterium signal in many lock
solvents is well known. Therefore the difference in resonance frequency of the lock
solvent and TMS is also known. As a consequence, TMS does not need to be added to
the sample to set =0; the spectrometer can use the lock frequency to calculate =0.

8.4 Shim Coils

The purpose of shim coils on a spectrometer is to correct minor spatial
inhomogeneities in the Bo magnetic field. These inhomogeneities could be caused by
the magnet design, materials in the probe, variations in the thickness of the sample
tube, sample permeability, and ferromagnetic materials around the magnet. A shim
coil is designed to create a small magnetic field which will oppose and cancel out an
inhomogeneity in the Bo magnetic field.
Because these variations may exist in a variety of functional forms (linear, parabolic,
etc.), shim coils are needed which can create a variety of opposing fields.
By passing the appropriate amount of current through each coil a homogeneous B o
magnetic field can be achieved. The optimum shim current settings are found by either
minimizing the linewidth, maximizing the size of the FID, or maximizing the signal
from the field lock. On most spectrometers, the shim coils are controllable by the
computer. A computer algorithm has the task of finding the best shim value by
maximizing the lock signal.
8.5 Sample Probe

The sample probe is the name given to that part of the spectrometer which accepts the
sample, sends RF energy into the sample, and detects the signal emanating from the
sample. It contains the RF coil, sample spinner, temperature controlling circuitry, and
gradient coils. The RF coil and gradient coils will be described in the next two
sections. The sample spinner and temperature controlling circuitry will be described
here.

The purpose of the sample spinner is to rotate the NMR sample tube about its axis. In
doing so, each spin in the sample located at a given position along the Z axis and
radius from the Z axis, will experience the average magnetic field in the circle defined
by this Z and radius. The net effect is a narrower spectral linewidth. To appreciate this
phenomenon, consider the following examples.
Picture an axial cross section of a cylindrical tube containing sample. In a very

homogeneous Bo magnetic field this sample will yield a narrow spectrum. n a more
inhomogeneous field the sample will yield a broader spectrum due to the presence of
lines from the parts of the sample experiencing different B o magnetic fields. When the
sample is spun about its z-axis, in homogeneities in the X and Y directions are
averaged out and the NMR line width becomes narrower.
Many scientists need to examine properties of their samples as a function of

temperature. As a result many instruments have the ability to maintain the
temperature of the sample above and below room temperature. Air or nitrogen which
has been warmed or cooled is passed over the sample to heat or cool the sample. The
temperature at the sample is monitored with the aid of a thermocouple and electronic
circuitry maintains the temperature by increasing or decreasing the temperature of the
gas passing over the sample.
8.6 RF Coils
RF coils create the B1 field which rotates the net magnetization in a pulse sequence.
They also detect the transverse magnetization as it precesses in the XY plane. Most
RF coils on NMR spectrometers are of the saddle coil design and act as the
transmitter of the B1 field and receiver of RF energy from the sample. You may find
one or more RF coils in a probe.

Each of these RF coils must resonate, that is they must efficiently store energy, at the
Larmor frequency of the nucleus being examined with the NMR spectrometer. All
NMR coils are composed of an inductor, or inductive elements, and a set of capacitive
elements. The resonant frequency, , of an RF coil is determined by the inductance
(L) and capacitance (C) of the inductor capacitor circuit.
RF coils used in NMR spectrometers need to be tuned for the specific sample being
studied. An RF coil has a bandwidth or specific range of frequencies at which it
resonates. When you place a sample in an RF coil, the conductivity and dielectric
constant of the sample affect the resonance frequency. If this frequency is different
from the resonance frequency of the nucleus you are studying, the coil will not
efficiently set up the B1 field nor efficiently detect the signal from the sample. You
will be rotating the net magnetization by an angle less than 90 degrees when you think
you are rotating by 90 degrees. This will produce less transverse magnetization and
less signal. Furthermore, because the coil will not be efficiently detecting the signal,
your signal-to-noise ratio will be poor.
The B1 field of an RF coil must be perpendicular to the B o magnetic field. Another

requirement of an RF coil in an NMR spectrometer is that the B 1 field needs to be

homogeneous over the volume of your sample. If it is not, you will be rotating spins
by a distribution of rotation angles and you will obtain strange spectra.
8.7 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.
Since the vertical bore superconducting magnet is most common, the gradient coil
system will be described for this magnet.
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 B o 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 coils.
The Y axis figure-8 coils provides a similar gradient in Bo along the Y axis.
8.8 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. Frequency and o 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 artifacts in the
spectrum. One is called a DC offset artifact and the other is called a quadrature
artifact.
8.9 Digital Filtering
Many newer spectrometers employ a combination of oversampling, digital filtering,

and decimation to eliminate the wrap around artifact. 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. The following flowchart summarizes the
effects of the three steps by showing the result of performing an FT after each step.


9. POSITION OF SIGNALS (CHEMICAL SHIFT)
In (NMR), the chemical shift describes the dependence of nuclear magnetic energy
levels on the electronic environment in a molecule.
An atomic nucleus can have a magnetic moment (nuclear spin), which gives rise to
different energy levels and resonance frequencies in a magnetic field. The total
magnetic field experienced by a nucleus includes local magnetic fields induced by
currents of electrons in the molecular orbitals (note that electrons have a magnetic
moment themselves). The electron distribution of the same type of nucleus (e.g. 1H,
13
C, 15N) usually varies according to the local geometry (bond lengths, angles between
bonds ...), and with it the local magnetic field at each nucleus. This is reflected in the
spin energy levels (and resonance frequencies). The variation of nuclear magnetic
resonance frequencies of the same kind of nucleus, due to variations in the electron
distribution is called the chemical shift. The size of the chemical shift is given with
respect to a reference frequency or reference sample, usually a molecule with a barely
distorted electron distribution.
9.1 The induced magnetic field
The electrons around a nucleus will circulate in a magnetic field and create a
secondary induced magnetic field. This field opposes the applied field and the nucleus
is therefore said to be shielded. Trends in chemical shift are explained based on the
degree of shielding or deshielding. E.g. of deshielding effect: the hydrogen with a
Electron withdrawing atom or group nearby.
Nuclei are found to resonate in a wide range to the left (or more rare to the right) of
the internal standard. When a signal is found with a higher chemical shift:
• the applied effective magnetic field is lower, if the resonance frequency is

fixed, (as in old traditional CW spectrometers)
• the frequency is higher, when the applied magnetic field is static, (normal case
in FT spectrometers)
• the nucleus is more deshielded
• the signal or shift is downfield or at low field or paramagnetic

Conversely a lower chemical shift is called a diamagnetic shift, and is upfield and
more shielded.
9.2 Operating frequency
The operating frequency ω0 of a magnet is calculated from the Larmor equation
where B0 is the actual strength of the magnet in units like teslas or gauss, and γ is the
gyromagnetic ratio of the nucleus being tested which is in turn calculated from its
magnetic moment μ and spin number I with the nuclear magnet μN and the Planck
constant h:
Thus, the proton operating frequency for a 1 T magnet is calculated as:
9.3 Chemical shift referencing
Chemical shift δ is usually expressed in parts per million (ppm) by frequency, because
it is calculated from:
Since the numerator is usually in hertz, and the denominator in megahertz, delta is
expressed in ppm.

The detected frequencies (in Hz) for 1H, 13C, and 29Si nuclei are usually referenced
against TMS (tetramethylsilane), which is assigned the chemical shift of zero. Other
standard materials are used for setting the chemical shift for other nuclei.
Thus, an NMR signal at 300 Hz from TMS at an applied frequency of 300MHz has a
chemical shift of:
Although the frequency depends on the applied field the chemical shift is independent
of it. On the other hand the resolution of NMR will increase with applied magnetic
field resulting in ever increasing chemical shift changes.

10.1H NMR SOLVENTS
NMR spectra are usually obtained by dissolving the sample in an appropriate solvent.
Because the solvent is present in much higher concentration than the sample, protons
in the solvent would overwhelm the resonance peaks for the sample. For this reason,
deuterated solvents are used in NMR spectroscopy. Completely deuterated solvents
are very difficult to make and expensive. In almost all cases, residual protons in the
deuterated solvent will cause a resonance peak (or peaks) to appear in the spectrum.
The following table shows the chemical shift values for the more common solvents
used for 1H NMR spectroscopy.
Solvent  (ppm)
acetic acid - d4 2.0 (singlet), 11.7(singlet)
acetone - d6 2.09 (singlet)
acetonitrile - d3 1.93 (singlet)
benzene - d6 7.15 (singlet)
carbon tetrachloride none
chloroform - d 7.25 (singlet)
dimethylsulfoxide - d6 2.49 (singlet)
ethanol - d6 1.11, 3.56, 5.19
methanol - d4 3.31 (singlet), 4.78 (singlet)
methylene chloride - d2 5.32 (singlet)
water - d2 4.82 (singlet)

11. FACTORS INFLUENCING CHEMICAL SHIFT
11.1 Electronegativity Effects
Electronegative elements directly attached to a carbon atom bearing hydrogens

(protons) pull electron density away from the protons. These protons are "deshielded"
because the valence electrons normally "shielding" these protons from the external
magnetic field are being inductively drawn away from those protons by the presence
of the electronegative element. The following table shows how chemical shifts for
methyl protons are affected by the presence of an electronegative element.
Compound CH3X CH3H CH3I CH3Br CH3Cl CH3F
Element X H I Br Cl F
Electronegativity of X 2.1 2.5 2.8 3.1 4.0
Chemical Shift  0.23 2.16 2.68 3.05 4.26
11.2 Hybridization Effects
11.2.1 sp3 Hydrogens
Hydrogens attached to sp3 hybridized carbon atoms resonate between 0 - 2 ppm.
Figure. sp3 hydrogens.

11.2.2 sp2 Hydrogens
Hydrogens attached to sp2 hybridized carbon atoms resonate farther downfield than
for normal aliphatic protons. The shift from TMS is dependent on the type of sp2
hybridized carbon atom:
Vinylic Hydrogens
Hydrogens attached to carbon-carbon double bonds resonate between 4.5 - 7 ppm.

The sp2 hybridized carbon atom of the double bond has increased s-character, and is
therefore more electronegative than an sp3 hybridized carbon atom.
Aromatic Hydrogens
Aromatic hydrogens resonate between 7 - 8 ppm.
Aldehyde Hydrogens
Aldehyde protons resonate between 9 - 10 ppm. This further downfield shift is due to
the additional effect of the electron withdrawing oxygen atom nearby.
11.2.3 sp Hydrogens
Acetylenic hydrogens resonate between 2 - 3 ppm due to the anistropy of the carbon-
carbon triple bond.
11.3 Hydrogen Bonding Effects
Hydrogen bonding causes a further de-shielding of protons, and a further downfield

shift for these proton resonances. Hydrogen bonding effects are concentration and
temperature dependent, and these results in a wide range of possible resonance
frequencies for these protons. In general, protons attached to oxygen and nitrogen
resonate between 0.5 - 5 ppm.

11.4 Magnetic Anisotropy Effects
All groups in a molecule with pi electrons will have an effect on the local magnetic
field due to the induced circulation of these pi electrons.
Aromatic Rings
The pi electrons in an aromatic ring are induced to circulate around the ring in
response to an applied magnetic field. This "ring current" generates a local magnetic
field which opposes the applied magnetic field. However, on the periphery of the ring,
the flux lines are in the direction of the applied magnetic field. Consequently, protons
attached to the aromatic ring "feel" a larger magnetic field than protons elsewhere in
the molecule. Aromatic ring protons will therefore resonate at higher frequency and
exhibit a downfield shift (7 - 8 ppm).
Figure . Magnetic anisotropy of an aromatic ring and that of a C,C-triple bond.
Acetylenic Hydrogens
The  electrons in a triple bond circulate around the bond axis to produce a magnetic
field directly opposing the applied magnetic field. The acetylenic hydrogen is shielded
by this induced field, and will therefore resonate at lower frequency (2 -3 pmm).

12.SPIN-SPIN COUPLINGS (SPLITTING OF SIGNALS)
The chemical shift is not the only indicator used to assign a molecule. Because nuclei
themselves are little magnets they influence each other, changing the energy and
hence frequency of nearby nuclei as they resonate—this is known as spin-spin
coupling. The most important type in basic NMR is scalar coupling. This interaction
between two nuclei occurs through chemical bonds, and can typically be seen up to
three bonds away.
The effect of scalar coupling can be understood by examination of a proton which has
a signal at 1ppm. This proton is in a hypothetical molecule where three bonds away
exists another proton (in a CH-CH group for instance), the neighboring group (a
magnetic field) causes the signal at 1 ppm to split into two, with one peak being a few
hertz higher than 1 ppm and the other peak being the same number of hertz lower than
1 ppm. These peaks each have half the area of the former singlet peak. The magnitude
of this splitting (difference in frequency between peaks) is known as the coupling
constant. A typical coupling constant value would be 7 Hz.
The coupling constant is independent of magnetic field strength because it is caused

by the magnetic field of another nucleus, not the spectrometer magnet. Therefore it is
quoted in hertz (frequency) and not ppm (chemical shift).
In another molecule a proton resonates at 2.5 ppm and that proton would also be split
into two by the proton at 1 ppm. Because the magnitude of interaction is the same the
splitting would have the same coupling constant 7 Hz apart. The spectrum would have
two signals, each being a doublet. Each doublet will have the same area because both
doublets are produced by one proton each.
The two doublets at 1 ppm and 2.5 ppm from the fictional molecule CH-CH are now
changed into CH2-CH:
• The total area of the 1 ppm CH2 peak will be twice that of the 2.5 ppm CH
peak.

• The CH2 peak will be split into a doublet by the CH peak—with one peak at 1
ppm + 3.5 Hz and one at 1 ppm - 3.5 Hz (total splitting or coupling constant is
7 Hz).
In consequence the CH peak at 2.5 ppm will be split twice by each proton from the
CH2. The first proton will split the peak into two equal intensities and will go from
one peak at 2.5 ppm to two peaks, one at 2.5 ppm + 3.5 Hz and the other at 2.5 ppm -
3.5 Hz—each having equal intensities. However these will be split again by the
second proton. The frequencies will change accordingly:
• The 2.5 ppm + 3.5 Hz signal will be split into 2.5 ppm + 7 Hz and 2.5 ppm
• The 2.5 ppm - 3.5 Hz signal will be split into 2.5 ppm and 2.5 ppm - 7 Hz
The net result is not a signal consisting of 4 peaks but three: one signal at 7 Hz above
2.5 ppm, two signals occur at 2.5 ppm, and a final one at 7 Hz below 2.5 ppm. The
ratio of height between them is 1:2:1. This is known as a triplet and is an indicator
that the proton is three-bonds from a CH2 group.
This can be extended to any CHn group. When the CH2-CH group is changed to CH3-
CH2, keeping the chemical shift and coupling constants identical, the following
changes are observed:
• The relative areas between the CH3 and CH2 subunits will be 3:2.
• The CH3 is coupled to two protons into a 1:2:1 triplet around 1 ppm.
• The CH2 is coupled to three protons.
Something split by three identical protons takes a shape known as a quartet, each
peak having relative intensities of 1:3:3:1.
A peak is split by n identical protons into components whose sizes are in the ratio of
the nth row of Pascal's triangle:
n
0 singlet 1
1 doublet 1 1
2 triplet 1 2 1
3 quartet 1 3 3 1

4 pentet 1 4 6 4 1
5 sextet 1 5 10 10 5 1
6 septet 1 6 15 20 15 6 1
7 octet 1 7 21 35 35 21 7 1
8 nonet 1 8 28 56 70 56 28 8 1
Because the nth row has n+ 1 component, this type of splitting is said to follow the
"n+1 rule": a proton with n neighbors appears as a cluster of n+ 1 peak.
With 2-methylpropane, (CH3)3CH, as another example: the CH proton is attached to

three identical methyl groups. The C-H signal in the spectrum would be split into ten
peaks according to the (n + 1) rule of multiplicity. Below are NMR signals
corresponding to several simple multiplets of this type. Note that the outer lines of the
nonet (which are only 1/8 as high as those of the second peak) can barely be seen,
giving a superficial resemblance to a septet.
When a proton is coupled to two different protons, then the coupling constants are
likely to be different, and instead of a triplet, a doublet of doublets will be seen.
Similarly, if a proton is coupled to two other protons of one type, and a third of
another type with a different coupling constant, then a triplet of doublets is seen. In
the example below, the triplet coupling constant is larger than the doublet one. The
analysis of such multiplets (which can get very much more complicated than the ones

shown here) provides important clues to the structure of the molecule being studied.
It should be emphasized that the simple rules for the spin-spin splitting of NMR
signals described above only apply if the chemical shifts of the coupling partners are
substantially larger that the coupling constant between them, otherwise there may be
more peaks, and the intensities of the individual peaks will be distorted (second-order
effects).

13.COUPLING CONSTANTS
The distance between peaks in a multiplet is called the coupling constant, J. For the
peaks of chloroethane, the coupling constant J is 7.2 Hz. This is calculated in the
following way: take the distance (in ppm) between any two adjacent split peaks. In
our example, the triplet centered at 1.14 ppm was used. The leftmost peak resonates at
1.26 ppm. The center peak resonates at 1.14 ppm. The difference between the two is
0.12 ppm. This spectrum was obtained on a 60 MHz machine. Therefore, there are 60
Hz for every 1 ppm on the spectrum. Multiplying 0.12 ppm by 60 Hz/1 ppm gives the
7.2 Hz value.
distance between split peaks = 1.26 ppm - 1.14 ppm = 0.12 ppm
J = 0.12 ppm * (60 Hz / 1 ppm) = 7.2 Hz
Peaks for protons that split each other will always have the same coupling constant.
This is useful in determining which peaks are related to each other in terms of
adjacency. One final note about peak splitting: multiplets will often be skewed in the
direction of the peak to which they are coupled.
The following table gives typical coupling constants for various functional groups.
Typical coupling constants for various functional groups.
Functional Group J (Hz)
6-8
11 - 18
6 - 15
4 - 10

6 - 10
8 - 11
a,a: 8 – 14
a,e: 0 – 7
e,e: 0 - 5
cis: 6 – 12
trans: 4 - 8
5-7

14. INTERPRETING NMR SPECTRA
There is no ONE single strategy for interpreting an NMR spectrum and correlating it
to a compound. Spectral interpretation is more like solving a puzzle, and the
following approach is merely one method. You may formulate, after many
interpretations, another method that you feel more comfortable using.
What does my compound look like?
Draw as accurate a picture of your compound as possible.
What nucleus am I observing?
If you acquire data on more than one nucleus, make sure you know which spectrum
belongs to which nucleus.
How many resonances do I expect to see in the absence of any coupling?

Determine which groups are chemically equivalent/in-equivalent. Does your
compound have a mirror plane that relates nuclei? A mirror plane does not always
exist, but for symmetrical molecules (such as a mono substituted benzene) a mirror
plane often is present.
Where do I expect to observe these resonances, i.e. what is their expected

chemical shift? The chemical shift depends on the local environment. Peaks far (9-12

ppm) from TMS are usually unique and identifyable. Peaks close to TMS (0-1 ppm)
are usually methyl groups. Typical chemical shifts are given below:
• Acids and aldehydes appear between 12-9 ppm. Both aldehydes and acids
appear in that region, although for different reasons (right?). To differentiate
between acids and aldehydes it is possible to exchange the acid proton with
deuterium and make it disappear. Often it is enough to simply add D2O to the
sample. The acid signal disappears and a new signal around 5 ppm for DOH
appears. Aldehydes do not exchange and so we can tell the difference between
the two groups. This technique is very useful for many functional groups where
the proton is attached to a heteroatom, especially oxgyen. Alcohols, phenols,
and carboxylic acids exchange rapidly and D2O is often sufficient for the
exchange process. Amines and thiols need stronger bases, such as NaOD, to
force the process to take place.
• Arenes and Alkenes occur near 8-5 ppm. Compounds with sp2 C-H fragments
fall into this region. Unless the alkene has a strongly electron withdrawing
group attached to the double bond, the alkene protons will absorb between 5-6
ppm. The integrated intensity can provide useful hints to discern between
alkenes and arenes. Alkenes have at most three protons while arenes can easily
have 4-5 protons. To confirm the assignment we need to look carefully at the
spin spin pattern.
• Aliphatic C-H protons appear between 5-0 ppm. Aliphatic fragments adjacent to
heteroatoms or electron withdrawing groups fall into the 2-5 ppm region.
Saturated C-H fall into the 2-0 ppm region. If you attach 2 electron
withdrawing groups to the carbon, the proton may appear in the 5-6 ppm region
where alkenes normally show up. Most organic texts will have tables of
common shifts for specific functional groups. Use these tables to make proton
assignments. Often, coupling NMR with IR to obtain information about the
functional groups that are present is a very good idea.
• Possible problems that may appear: (i) some protons can appear over wide ppm
ranges, mostly N and O bound protons, making identification difficult; and (ii)
the appearance of stray peaks from your solvent (or the solvent used to clean
the NMR tube) or an impurity.

What is the integral value or ratio between integral values for each resonance?
Integration values depend on the amount of each kind of nucleus present. Within one
given molecule, the ratio between chemically inequivalent nuclei is set, and
inspection of the integral values or ratios must follow directly from the molecular
formula. Integration can only be compared for resonances arising from the same
molecule. For two different molecules, the integral values for similar groups will be
quite different if the concentration of the molecules is different!
If you normalize an integral value for a resonance that you can clearly identify, the
other integral values can be compared directly. For instance, normalizing a clearly
identified CH3 resonance will result in a -CH2- integral value of 2, a C5H5 integral
value of 5, etc., for proton fragments on the same molecule only.
If you cannot clearly identify a resonance as belonging to a particular group, a

comparison of the ratios of the integral values is necessary. For instance, the ratio of
CH3/CH2 integral values should equal 1.5, and the ratio of CH3/C5H5 should equal 0.6,
etc., for proton fragments on the same molecule only.
What is the expected multiplicity of each resonance?
Multiplicity can be calculated from M = 2nI + 1 where n = # of equivalent nuclei on

adjacent carbons and I = nuclear spin. For the common nuclei that you may
encounter, such as 1H, 13C, 19F, 31P, and 195
Pt, the nuclear spin is ½ and the above
formula collapses to M = n + 1. For a doublet you have 1 adjacent proton, for a triplet
you have 2 adjacent protons, and for a quadruplet you have 3 adjacent protons. Some
splitting patterns can become very complicated, such as for aromatic protons, and may
be difficult to determine. Multiplicity is very important in determining neighboring
spin-active nuclei.
Assemble the fragments into a molecule.
Compare your assembled molecule to the molecule you initially drew in Step 1 and
rearrange the connectivity as necessary.

Calculate the coupling values, if present.
Coupling is a pairwise relationship. In a simple case of one proton coupling to only

one other proton, you MUST observe TWO DOUBLETS that have identical J values.
If you observe coupling between a proton and fluorine, JHF will be observed in the 1H
NMR (one member of the pair) and JHF will also be observed in the 19F NMR (the
other member of the pair). You may observe the pairwise relationship in one NMR
spectrum (homonuclear coupling) or between two different spectra (heteronuclear
coupling). This the reason for Step 2.
Coupling is a measure of the degree of interaction between two nuclei, and is

represented by the letter J (reported in units of Hz). In general, 1J > 2J > 3J, where the
superscript refers to the number of bonds between the coupled nuclei. 4J is generally
not observed in routine spectra. 2,3J typically ranges from 2-20 Hz.
To calculate J, you will need to know the scan frequency, or SF (PCNMR refers to
this as "synthesizer frequency" in the parameter list).
Different nuclei will have different values of SF, and you must check the parameter
list to obtain the exact value of SF for your particular experiment before you attempt
to calculate J.
nucleus SF (MHz)
1
H 300
31
P ∼ 121
13
C ∼ 75
Trace through your fragment assembly, comparing it to your the molecule you
initially drew.

If you assembled your fragments correctly, the multiplicity, coupling, and chemical
shifts should match and in all likelihood you have correctly interpreted your NMR
spectrum. If there are inconsistencies, you must (obviously) try again with another
assembly of fragments.

15. NMR APPLICATIONS
Today, NMR has become a sophisticated and powerful analytical technology that has
found a variety of applications in many disciplines of scientific research,
medicine, and various industries. Modern NMR spectroscopy has been
emphasizing the application in biomolecular systems and plays an important
role in structural biology. With developments in both methodology and
instrumentation in the past two decades, NMR has become one of the most
powerful and versatile spectroscopic techniques for the analysis of
biomacromolecules, allowing characterization of biomacromolecules and
their complexes up to 100 kDa. Together with X-ray crystallography, NMR
spectroscopy is one of the two leading technologies for the structure
determination of biomacromolecules at atomic resolution. In addition, NMR
provides unique and important molecular motional and interaction profiles
containing pivotal information on protein function. The information is also
critical in drug development. Some of the applications of NMR spectroscopy
are listed below:
• Solution structure The only method for atomic-resolution structure

determination of biomacromolecules in aqueous solutions under near
physiological conditions or membrane mimeric environments.
• Molecular dynamics The most powerful technique for quantifying motional
properties of biomacromolecules.
• Protein folding The most powerful tool for determining the residual structures
of unfolded proteins and the structures of folding intermediates.
• Ionization state The most powerful tool for determining the chemical
properties of functional groups in biomacromolecules, such as the ionization
states of ionizable groups at the active sites of enzymes.
• Weak intermolecular interactions Allowing weak functional interactions
between macrobiomolecules (e.g., those with dissociation constants in the
micromolar to millimolar range) to be studied, which is not possible with other
technologies.
• Protein hydration A power tool for the detection of interior water and its
interaction with biomacromolecules.

• Hydrogen bonding A unique technique for the DIRECT detection of hydrogen

bonding interactions.
• Drug screening and design Particularly useful for identifying drug leads and
determining the conformations of the compounds bound to enzymes,
receptors, and other proteins.
• Native membrane protein Solid state NMR has the potential for determining
atomic-resolution structures of domains of membrane proteins in their native
membrane environments, including those with bound ligands.
• Metabolite analysis A very powerful technology for metabolite analysis.
• Chemical analysis A matured technique for chemical identification and
conformational analysis of chemicals whether synthetic or natural.
• Material science A powerful tool in the research of polymer chemistry and
physics.

Nuclear Magnetic Resonance Spectros

Diunggah oleh

Informasi Dokumen

Deskripsi Asli:

Judul Asli

Hak Cipta

Format Tersedia

Bagikan dokumen Ini

Bagikan atau Tanam Dokumen

Opsi Berbagi

Apakah menurut Anda dokumen ini bermanfaat?

Apakah konten ini tidak pantas?

Hak Cipta:

Format Tersedia

Nuclear Magnetic Resonance Spectros

Diunggah oleh

Hak Cipta:

Format Tersedia

H1-Nuclear Magnetic Resonance Spectroscopy MPY-102

2. SPIN PROPERTIES OF NUCLEI

School of Pharmacy, SRTMU Nanded Page 1 of 41

Natural % Magnetic Magnetogyric

3. FEATURES THAT LEAD TO THE NMR PHENOMENON:

1. A spinning charge generates a magnetic field, as shown by the animation on the

School of Pharmacy, SRTMU Nanded Page 2 of 41

School of Pharmacy, SRTMU Nanded Page 3 of 41

the direct correlation of frequency (energy difference) with magnetic moment (h =

4. NUCLEAR SPIN AND THE SPLITTING OF ENERGY LEVELS

Subatomic particles (electrons, protons and neutrons) can be imagined as spinning on

School of Pharmacy, SRTMU Nanded Page 4 of 41

5. CALCULATING TRANSITION ENERGY

The constant, γ, is called the magnetogyric ratio and is a fundamental nuclear

The energy of a particular energy level is given by;

Where B is the strength of the magnetic field at the nucleus.

School of Pharmacy, SRTMU Nanded Page 5 of 41

There are two major relaxation processes;

• Spin - lattice (longitudinal) relaxation

6.1 Spin - lattice relaxation

School of Pharmacy, SRTMU Nanded Page 6 of 41

6.2 Spin - spin relaxation

School of Pharmacy, SRTMU Nanded Page 7 of 41

7.1 Protons with identical chemical environments:

School of Pharmacy, SRTMU Nanded Page 8 of 41

We are now in a position to measure experimentally the frequency of the process

School of Pharmacy, SRTMU Nanded Page 9 of 41

that we measure only the xy component magnetisation of M, and NOT absorption or

7.2. Protons with different electronic environments.

School of Pharmacy, SRTMU Nanded Page 10 of 41

School of Pharmacy, SRTMU Nanded Page 11 of 41

School of Pharmacy, SRTMU Nanded Page 12 of 41

8.3 Field Lock

School of Pharmacy, SRTMU Nanded Page 13 of 41

School of Pharmacy, SRTMU Nanded Page 14 of 41

8.4 Shim Coils

8.5 Sample Probe

School of Pharmacy, SRTMU Nanded Page 15 of 41

Picture an axial cross section of a cylindrical tube containing sample. In a very

Many scientists need to examine properties of their samples as a function of

School of Pharmacy, SRTMU Nanded Page 16 of 41

The B1 field of an RF coil must be perpendicular to the B o magnetic field. Another

School of Pharmacy, SRTMU Nanded Page 17 of 41

8.7 Gradient Coils

Assuming the standard magnetic resonance coordinate system, a gradient in Bo in the

School of Pharmacy, SRTMU Nanded Page 18 of 41

8.8 Quadrature Detector

School of Pharmacy, SRTMU Nanded Page 19 of 41

rotating frame of reference converter. The heart of a quadrature detector is a device

8.9 Digital Filtering

Many newer spectrometers employ a combination of oversampling, digital filtering,

School of Pharmacy, SRTMU Nanded Page 20 of 41

School of Pharmacy, SRTMU Nanded Page 21 of 41

9. POSITION OF SIGNALS (CHEMICAL SHIFT)

9.1 The induced magnetic field

• the applied effective magnetic field is lower, if the resonance frequency is

School of Pharmacy, SRTMU Nanded Page 22 of 41

9.2 Operating frequency

The operating frequency ω0 of a magnet is calculated from the Larmor equation

Thus, the proton operating frequency for a 1 T magnet is calculated as:

9.3 Chemical shift referencing

School of Pharmacy, SRTMU Nanded Page 23 of 41