Most spinning nuclei behave like magnets.

However, as opposed to the behavior of a classical magnet the

nuclear spin magnetic moment does not always align with the
direction of the magnetic field B (quantization). NMR
fundamentals are the same for all magnetic nuclei.
Nuclear Magnetic Resonance
Spectroscopy
0
Basics
0

In the absence of a field the nuclear magnets orient in all

possible directions in space.

Nuclear spin quantum number I Precessional movement

Depending on the nuclear constitution, they would possess a
nuclear spin moment.
Bare proton
0
A Z I (nuclear spin q.n.)
Mass # Atomic #
Odd Odd or even n/2 (n 0 & odd integer) I =1/2
Even Even 0
2 =
Even Odd n (integer)
magnet of magnetic moment (dipole) 0

Nuclei of interest 1H, 2H, 13C, 19F, 29Si, 31P, 127I, etc.
Any nucleus with an odd Z or an odd A has a net spin.
Nuclear angular momentum of 0 0 = I ( I + 1) =
0 = 0 = I ( I + 1) =
magnetogyric ratio of the nucleus
http://www.chem.queensu.ca/FACILITIES/NMR/nmr/webcourse/index.htm
http://www.eclipse.net/~numare/nsinmrpt.htm
 # orientations (states) in a B =2I+1 Precessional cone
Each state characterized by mI = -I, -I+1,.+I Of a single
Z, B0
nucleus 0 0, dipole
moment
I = 1/2
mI = +1/2; , ground state m = m I =
m
mI= -1/2 0 = B0 Vertical component
remains steady.
B 54.7o
Horizontal component
Energy

Y
changes direction.

X I=1/2 Ground state

mI= +1/2
mI = -1/2; , excited state
0= Larmor frequency, rad/s
In the presence of a B the two states differentiates. The quantity NMR experiment measures.

mI = +1
mI = + , low energy

I=1/2 mI= 0

I=1
mI = - 1/2

http://www.bruker-nmr.de/guide/eNMR/chem/NMRnuclei.html mI= -1

mI=+3/2
Transitions
I=3/2
between states
Selection rule: dictated by
mI=1 selection rules
or

mI= - 3/2
 Energy Changes in a varying B
1
1 E = [+ =]B
+ z = + = 2
2 m= -1/2, state
E anti-parallel to field,
B
1
E = [ =]B
1 0,0 2
z = = 1
2 E = [ =]B B
2 B0

m= +1/2, state
E = =B B , parallel to field,
1
E = [+ =]B
2
magnetogyric ratio of the nucleus z = mI = Z
E = B
E = =B0 = h
falls in the radio frequency region.

Continuous wave experiment; Field sweep

NMR Experiment; pseudo classical frequency constant

E = =B0 = h 0
E B0
0 =
0,0 2
B
0
B0

B
B E = =B0 = h 0
B
B0
Continuous wave experiment; Frequency sweep Proton in a sample are in a different electronic environment
B constant than a bare proton. Protons with different type of environments,
characterized by , can be recognized.

E Each type of proton has the same environment if free rotation is

possible.
0,0
BE = =B0 = h 0
B0 B0
0 = O a
2 H3C
CH3
B c
a b c b

0

In the NMR experiment, the nuclear energy levels are excited

and the absorptions of the nuclear excitations recorded.

Note the hyperfine splitting of NMR peaks.

Bopp Bopp shields Increasing (frequency sweep)
the nucleus
Screening constant Increasing , B (field sweep)

High

Bopp = B0 Low Field H

Field
H

depends on the electron

B = (1 ) B0 Bo
density around the atom.

(1 ) B0 Therefore of absorption
= of each type of H is unique.
2
 E
N
The pseudo NMR experiment is impractical, i.e. absorbance =e kT
cannot be measured directly by experiment. N
The reason for it is a population distribution issue and the N E E
nature of the interaction of electromagnetic radiation with = 1 Using Taylor series for << 1
matter (energy levels of matter).
N kT kT

N E
very small N N E
Boltzmann Law
=e

kT =
N N kT
E
N N = N
For protons = 26.75 107 rad/T s; h-bar = 1.055 10-34 Js, kT
k = 1.380658 10-23 J/K. At 298K and B0 = 7.05T
Population excess is linear in E
N under this condition.
= 0.99995 1 N N
N

Samples contain millions of molecules. All nuclei of the same Relationship between energy levels,
type would precess at the same Larmor frequency. magnetic field and shielding:

The vector sum of the magnetic moments of one type of nuclei is Low , low shielding
a measure of the number of protons of that type in the molecule. High transition energy

High , high shielding

The determination of the magnetic moment for protons Low transition energy
in each type of electronic environment forms the essence of
the NMR experiment.

O Same B
Ma:Mb:Mc=3:2:3
H3C
CH3
(1 ) (1 )
a b c = B0 slope = 2 =
2 2
Chemical Shift raised _

The resonance frequency (shifts) depends on the value of

, the electronic environment around the nucleus,
_
Larger electron densities has smaller Larmour frequencies.
lowered
shielding
Factors determining the frequency shift: , Beff;

a. electro-negativity of the neighboring groups

b. electron currents (delocalization) in response to Bo
leading to magnetic fields which are anisotropic that
would enhance/diminish the effective field at the
nucleus .
(1 )Beff (1 )Beff
= =
2 2

Increasing (frequency sweep)

In FTNMR vocabulary a x MHz instrument would mean an
High , high shielding instrument where the applied field B0 is such that the
Low , low shielding
Low transition energy bare protons would rotate at a Larmor frequency of x MHz.
High transition energy
Increasing , B (field sweep)
High In such an instrument, the AC frequency of the pulse for
Field H
Low excitation of bare protons is x MHz.
Field
H
MSUM - 400MHz instrument.

The pulse generates a range of frequencies centered at

400MHz.
 1H 2H 13C 14N 15N

Nuclear Spin, I 1/2 1 1/2 1 1/2

Nuclear
magnetic +2.79284 +0.85743 +0.70241 0.40376 -0.28319
moment,
Gyromagnetic
Ratio 26.7510 x 10 7 4.1064 x 10 7 6.7263 x 10 7 1.9331 x 10 7 -2.7116 x 10 7
(rad T-1 s-1)
Quadrupole 1.6 x 10 -30
2.73 x 10 -31
moment (m2)
Relative 1.59 x 10 -2 1.01 x 10 -3 1.04 x 10 -3
1.00 9.65 x 10 -3
Sensitivity
Natural 1.10 99.63 0.37
99.985 0.015
Abundance, %
Absolute 1.00 5.69 0.0219
5680 8.2 x 10 -6
Sensitivity

(1 ) B0
=
2 B0
0 =
2

A bare 1H(=0) at 2T field resonates at 90.0000 MHz;

For bare protons = 0. = 26.75 107 rad T-1 s-1 and
B0 = 7.05T the Larmor frequency is;
A typical proton would have a frequency of 89.9995 MHz,
In the same field, it is an unwieldy number to report, further
in a different instrument (different field) the frequency will be
(1 ) B0 different.
=
2
Thus absorption positions are reported with respect to a
B0
= (1 ) reference.
2
= 3 108 Hz
General presentation of NMR spectra:
The position of NMR resonance signals (frequency scale) is
dependent on the external magnetic field strength, B0 (as
Increasing (frequency sweep) opposed to UV, IR, AA etc).

Increasing , B (field sweep) sample =H(1-sample)B0

High
Low Field B Since no two magnets in the spectrometers will have exactly
Field B the same field, resonance frequencies will vary accordingly.
A method for specifying uniquely the position of NMR signals
is needed.
Ref
To express the position of NMR signals unequivocally, the
signals are expressed relative to signal from a standard
compound.
0

Chemical shift

The reference compound is chosen so that the protons of it CH3

are well shielded than the typical protons in compounds of TMS
H3C Si CH3
interest (so that reference absorptions does not interfere with
the resonances normally observed for organic compounds . CH3

Further it should give a single sharp nmr signal. The most common reference compound used is
tetramethyl silane. TMS is rich in Hs(12) and methyl group Hs
It should be chemically un-reactive and easily removable from are richer in electrons (larger ) than in many organic compounds
the sample after the measurement. and they are structurally equivalent.

ref =H(1-ref)B0

sample- ref =H(ref- sample)B0

ref sample is a very small quantity.

 ref =H(1-ref)B0 Chemical Shift
sample reference
sample =H(1-sample)B0 = 10 6
rf
sample - ref =H(ref - sample)B0
sample reference
= 10 6
sample - ref =H(ref - sample) B0/2 rf
very small

Different kinds of protons usually appear at different chemical

shifts makes it possible to distinguish one kind of proton from Chemical shift is independent of the instrument field strength.
another. One reason for the different values of chemical shift
is the difference in shielding. If there is more electron density
around a proton, it sees a slightly lower magnetic field and
vice versa.

6 6

89.999510 89.999010
6
For bare 1H(=0)at 2T field resonate at 90.0000 MHz; Chemical Shift (ppm): 10 = 5.556
6
90 10
A typical proton would have a frequency of 89.9995 MHz,
it is an unwieldy number to report, further if a different
Frequency shift (Hz) 6 6
instrument (different field) is used the frequency will be 89.9995 10 89.9990 10 = 500
different.
Frequency shift in a 300MHz instrument (Hz):
Thus absorption positions are reported with respect to a 6
( 5.556) 300 10
reference. If a reference compound resonates at 89.9990MHz. = 1666.8
6
10
6 6
Chemical shift:
89.999510 89.999010
6 Entire proton range 300MHz instrument is 12ppm.
10 = 5.556
6
90 10
In (Hz): 6
( 12) 300 10
= 3600
6
10
Chemical shift decreases with increasing shielding of the nucleus The higher the field the larger the frequency range over which
the spectrum spreads. This enhances the resolution of peaks.
Low , low shielding
High transition energy

High , high shielding

Low transition energy

Same B

( ref )
ref = B
2

Frequency shift is dependent of the instrument field strength. The resonance frequencies of nuclei in similar environments
are the same.

same = the mean of the frequencies.

The necessity to look at a mean arises from the fact that in

general the spins of the nuclei in close proximity do influence
each other and hence their resonant frequencies.
100MHz
Such influences, termed as coupling (scalar coupling) results
500MHz
in the splitting of the spectral peaks. There are weakly coupled
and strongly coupled systems.

734Hz vs. 3670Hz (Benzene)

Scalar (Spin) Coupling Classification of Spin Systems

Most of the time the NMR spectrum consists of groupings of The chemical shifts of protons in a molecule can differ in
multiple lines (multiplets). This is due to 1H - 1H coupling (spin- various degrees.
spin splitting/J-coupling/scalar coupling). The nuclei which are
1 to 3 bonds away (especially) would feel the nuclear spin Depending on the extent of coupling (J) between the
state of each other via the electron spins of the intervening nuclei, the splitting pattern changes.
bonds. That generates the splitting of spectral lines.
Simple rules (n+1) and area ratios (Pascal triangle) is useful for
H Br H H weakly coupled systems (first order spectra).
| | | |
Cl C C Br Cl C C Br The spin system, in general, for example is referred to as AnBm
Geminal 2J | | | | Vicinal 3J (second order spectra) if the chemical shifts are quite close
H Cl Cl Br together, and AnXm (first order spectra) if the chemical shifts
are far apart relative to J.
Only magnetically nonequivalent nucleii couple.

Pople Notation
CH3 H CH
H3C 3 A6X
Each chemically different type of proton is given a capital letter; A3X2
A,B C M,N. X,Y,Z.
The number of protons in a group protons is indicated as a
subscript; A2, B3
Protons with similar values are assigned letters close to one
another in the alphabet; A,B,C; M,N; X,Y, Z. H
H3C OH A3X2
H AMX
Two protons same chemical shift but magnetically nonequivalent
is assigned, for example AA. H3C O H
+
N A3M2X2
AX, AM weakly coupled, AB strongly coupled pair of protons. O
-

Three protons very different in from each other (and therefore H3C
I A3B2X2
weakly coupled) would be like AMX, ABX, BX weakly coupled,
AB strongly coupled.
First-order Scalar Coupling (weakly coupled systems) Consider an AX system:

Coupling of spins lead to peak splitting (hyperfine structure). A molecule which contains a proton (HA) attached to a C, and
In first order spectra the splitting of the peaks follows a simple that this C is attached to another C carrying a proton (HX). In
pattern. this situation HA feels the presence of HX via bonding
H3C A6X electrons.
A H3C CH3
O
O
Protons are tiny little magnets, that orients either parallel or
anti-parallel the magnetic field B0 of the instrument. When the
field created by HX effectively reinforces the magnetic field B0
HA feels a slightly stronger field, but when the field created by
HX opposes B0, HA feels a slightly weaker field.

This leads to two signals for HA depending on the alignment of

HX. Similarly HX can feel either a slightly stronger or weaker
field due to HA.

13C-1H AX System 1J
(1 ) B0 HX HA
= | | Fermi contact Pauli Principle
2
Cl C C Br
| |
Cl Br

>8 High energy
J
A A X
X


Isolated atoms Fermi
contact energetically favors Low energy
anti-parallel orientation
of spins. Bonded atoms Pauli Principle
B0+BA B0-BA B0+BX B0-BX
demands anti-parallel electron
spins making nuclear and
3J 3J electron spins parallel, .
XA HX AX HA
 13C-1H AX System 1J 13C-1H AX System 1J

C
The difference in









the nuclear spin of
bonded atom (left) gave
rise to four eigen states.

If otherwise it


would be two eigen
states and one frequency. C





Eigen-states with no coupling Effect on eigen-states with coupling

H H
13C-1H AX System 1J AX System 3J
Cl Br
Cl Br
J/4

+ (J/4/ + J/4)

Fermi contact
C
J/4 J
A X
C C



Pauli Principle

J/4
- (J/4/ + J/4)
C
J/4 Hunds Rule

For the X peaks a similar situation (doublet) would form. High energy
 H H
Cl Br
AX System 3J

Fermi contact Cl Br
J/4

+ (J/4/ + J/4)

A X
X


J/4
Pauli Principle

Hunds Rule J/4

- (J/4/ + J/4)
Low energy X
J/4

For the X peaks a similar situation (doublet) would form.

H H AX System 2J AX System 1J
C

J/4
- (J/4/ + J/4)

X J/4
A A

X X
Regardless of sign J




the splitting pattern
would be the same for
J/4 same value of J.
High energy Low energy !!
+ (J/4/ + J/4)
X
J/4
J<0
Stick Diagram first order splitting due to scalar coupling Stick Diagram first order splitting
A
Spin System # Peaks

(n+1)

AX 2
J

AX2

doublet triplet quartet quintet 3

1:1 1:2:1 1:3:3:1 1:4:6:4:1
AX3
The splitting between peaks, J, are equal, the ratio of peaks
conform to the Binomial coefficients (first order spectra).
4
J is independent of the field strength of the instrument. Value of A
J (Hz) property of the spin system of the molecule. Centro-symmetric, equally spaced
peaks by J, with area ratios Binomial
coefficients!!.

Stick Diagram first order splitting Second order spectra

If the chemical shifts are
H similar then distortions in
A peak height could occur.
s For more than two spins,
extra signals may appear.
AM H
H These effects are observed
d in second order coupling.
H H
AMX
d-d
H
H H
AMXY
d-d-d
H H
First order splitting: Because chemical shift difference is dependent of the
>8 AX, AMX, AX 2 ,...
J magnetic field (actually increases with the applied field) and
the coupling constant is independent of the applied field, it is
possible to change the ratio;

Second order splitting: < 8 AB, ABC, AB2 ,...
J
J
= chemical shift difference of the coupling nuclei (Hz)
At larger fields would be larger while J remains the same.

Splitting pattern of the first order spectra are extremely useful Instruments with larger fields simplify the second order
To extract information about the connectivity and the pattern to a first order pattern.
stereochemistry of the molecule.

Coupling constant (J) has the same value regardless of the

60 MHz field strength.

Both homonuclear and heteronuclear coupling folow the

(n+1) rule.
300 MHz
J = f (bond angles, temperature and solvent -influence the
conformation of the compound)

1J > 2J > 3J ..; 1J , 3J, ... > 0 ; 2J, 4J <0

600 MHz

The appearance of the multiplet does not show the sign of J.

 H-NMR Experiment: One Pulse Experiment
Cl O O
P O Place the sample in the magnetic field.
Cl O OH
The nuclei attains equilibrium composition. Magnetic moments
orient on z-axis.

After a recycle delay, d1, apply a RF pulse (=/2).

The magnetic moments for each type of nuclei are moved onto
the x-y plane.
Note: Methylene protons
are magnetically non-equivalent.
The FID collected.
Each H 1:3:3:1 pattern &
are overlapped.
Caution: Overlapped peaks ratio
not Binomial coefficients!!. -CH2-

Z
Collection of nuclear
dipole magnetic
moments - symmetrical
components
Magnetic moment
vector, single B0
B0 nucleus
 Z
Z Z
Net vector of nuclear
dipole magnetic Tilt angle
moments, M0. M0
x
M0
B0 Y

X
X
Transceiver
Y
Use the Right Hand Rule.

Each M with its Larmor frequency generates a time varying

voltage at the receiver coil. Sine/cosine curve.
Y Y Y Y

X X X X
Y
t=0 T/4 T/2 3T/4 ..
X
Induced Voltage receiver mode
1D-H-NMR Experiment: One Pulse Experiment (Single Channel) Fourier Transformation of FID

Place the sample in the magnetic field.
The nuclei attains equilibrium composition. Net magnetic moments
orient on z-axis.
After a recycle delay, d1, apply a RF pulse (=/2).
The magnetic moments for each type of nuclei are moved onto
the x-y plane.
The FID collected (detector response).
=/2
Collect and add many FIDs.
The signal detected is a time domain signal. The signal
decays due to the T2 relaxation and is therefore called free
induction decay (FID). The equilibrium magnetization is
directed along the external magnetic field in Z). The time-
dependent signal can be converted into the frequency
domain by the Fourier transformation

The intensity of the detected signal with the highest value at 0

(resonance frequency)
p1

d1 AQ d1

H H
Cl Br
AX System 3J
Cl Br

For the X peaks a similar situation (doublet) would form.