Organic Spectroscopy

CHE 203 1.0

Prof. B. Asiri Perera

Department of Chemistry
University of Sri Jayewardenepura
NMR SPECTROSCOPY

- Basic Instrumentation -
1
 Principles of NMR
NMR is a spectroscopic technique
which is based on the absorption of
electromagnetic radiation in the radio
frequency region 4 to 900 MHz (long λ
and low E) by nuclei of the atom .

Used to characterize molecules by

identifying C-H fame work within the
molecule 2
All nuclei are charged because of
protons(P) and some of them behave as
if they spin.

This spinning charge generates a

magnetic field and behaves like a small
magnet.
These nuclear spins are
random in direction.
They can interact with external
magnetic field B0 (H0)

When a small B0
is applied nuclei align
themselves either with
or against the external
field 4
NMR phenomenon is not
observable in the absence of an
applied magnetic field,B0
A magnet is an essential
component of any NMR
spectrometer

5
P have a charge, P and N both have a
spin.
This spin can be +1/2 or -1/2
P=

N S

+1/2 -1/2
S N
To interact with B0 and to be NMR
active net spin and a charge must be
there in the nucleus. Net spin I
1H 1P, 1e
1 +1/2
+1/2

19F 9P, 10N, 9e

9

Odd mass number and odd atomic number.

Net spin present, NMR active.
2D 1P, 1N, 1e
1 Net spin I

+1/2 +1/2

Odd atomic number NMR active.

8
13C 6P, 7N, 6e
6 Net spin I

+1/2

Two identical partials can not have exactly

the same properties . In different E levels

Odd mass number NMR active.

When even number of P and N present no net
spin(magnetic movement cancel). Not interact
with B0. Even atomic and even mass number
NMR inactive.
Net spin I
12C 6P, 6N, 6e no net spin
6 0

16O 8P, 8N, 8e 0

8
Each spin active nucleus has a number
of spins defined by its nuclear
magnetic spin quantum number I
I can be equal to 0, 1/2 , 1, 3/2 ... etc.
12C 16O 0
6 8

1H 13C , 19F , 31P 1/2

1 6 9 15

2D 14N 1
1 7
Elements with either odd mass or odd atomic
number have nuclear spin and NMR active
Without With
external external
magnetic magnetic
field B0. field B0.

In a uniform magnetic field(B0), a nucleus of spin I

may assume 2I + 1 orientations.
1H 1
I=
2
𝟏
Number of orientations = ( 2 x ) + 1 = 2
𝟐
1 1
have clockwise (+ ) and counter clockwise (- ) spins.
2 2
 , ß

, α

B0
It is possible to induce a spectroscopic
transition (spin-flip/resonance) by the
absorption of electromagnetic energy (∆𝐄) of
the appropriate frequency ν (radio frequency)
B0
Higher B0
Frequency of the absorbed electromagnetic
radiation ν is not an absolute value for any
particular transition, but has a different value
depending on the strength of the applied
magnetic field, B0.
ν α B0
ν = k B0
Larmor equation
Stogner the B0 Higher the ν needed to
resonate. K is a constant characteristic of
the nucleus observed.
For every value of B0/H0 there is a matching value of 𝛎
corresponding to the condition of resonance according
to Equation

18
19
 1H NMR SPECTROSCOPY
NMR spectrum is a graph of intensity of
absorption of Rf radiation/NMR
signal(y-axis) vs frequency of the Rf
radiation (x-axis ).

Frequency scale, the units are converted

to ppm so that the scale has the same
numbers irrespective of the strength of
the B0 in which the measurement was made
C𝐇𝟑 − Y
Intencity

Low B0

Frequency
Intencity

Frequency high B0
The freq. difference between the resonance
of a nucleus and the resonance of the
reference compound is termed the chemical
shift.

Tetramethyl silane (TMS), (CH3)4Si is the

1 13
reference for both Hand C NMR. TMS is
added directly to the solvent of the
substance under study.
 Chemical Shift
chemical shift from TMS in Hz
chemical shift in ppm = 𝐬𝐩𝐞𝐜𝐭𝐫𝐨𝐦𝐞𝐭𝐞𝐫 𝐟𝐫𝐞𝐪𝐮𝐧𝐞𝐜𝐲 𝐢𝐧 𝐌𝐇𝐳

The chemical shift range for

1H --- 0-12 ppm (from TMS δ=0 )
13C --- 0-220 ppm (from TMS δ =0).

Chemical shifts can be measured in Hz but are

usually expressed in ppm.
 Advantages of Using TMS as a
Reference compound

It is a relatively inert liquid (low B.P

0
26.5 C) can be easily removed after
use
Gives a sharp single signal in both H 1
13
and C because the compound has
only one type of H and one type of C
.
The chemical environment of both C and H in
TMS is unusual due to the presence of Si and
signal occurs outside the range observed for
organic compounds, so the reference signal is
unlikely to overlap a signal from the
substance examined.

The chemical shift of TMS is not affected by

complexation or solvent effects because
TMS doesn't contain any polar groups.
Factors influence Chemical Shift

1.Deshieldingand shielding by
groups due to electro negative
atoms

2.Anisotropy due to magnetic

fields generated by π bonds
26
 Shielding and Deshielding
1H
If two environments are different
their chemical shifts are different.
CH3 CH2COH

If have identical molecular

environments they have the same
chemical shift, and termed chemically
equivalent. CH3 CO CH3
Nuclei tend to be deshielded by groups which
withdraw electron density(-OH, -OCOR, -OR,
-N02, halogen) .
Those deshielded nuclei can feel high B0 and
resonate at higher ppm values (away from
TMS).
High electron density around a nucleus
shields the nucleus from the external
magnetic field B0 and nuclei resonate at
lower ppm values (towards TMS).
28
Downfield / Deshield Upfield / shield

δ 12 δ6 δ0
ppm from TMS

R-C - H R-CH2-CH3

𝐁𝟎
 Anisotropy

Circulating π electrons generate secondary

magnetic field same way as B0(adding to B0) .
Deshield aromatic protons. Need higher radio
frequency to resonate. (appear around 7ppm)
secondary Deshield
magnetic
field

𝐁𝟎
Add to B0

π electrons generate secondary magnetic

field same way as B0(adding to B0) . Deshield
protons
 secondary
magnetic
Opposite to B0 Shield field
𝐁𝟎
π electrons generate secondary magnetic
field opposite to B0. shield protons. Feels less
B0. Need lower radio frequency to resonate
1H NMR Chemical Shifts
36
C=C-H
C=C-H

37
 Solvents for NMR Spectroscopy
NMR spectra are almost invariably obtained in
solution.
The most commonly used organic solvent is
deuterochloroform CDCl3. It has no protons
and has a deuterium atom.

For ionic and hydrophilic compounds the

common solvent is deuterated water, D2O.
Some other solvents used are CCl4 , C6D6 Hexa
deuteriobenzene, CS2
 Spin-Spin Coupling
A typical organic molecule contains more
than one magnetic nucleus (e.g. more than
one IH)
When one nucleus can sense the presence
of other nuclei through the bonds of the
molecule the signals will exhibit fine
structure (splitting or multiplicity).

Provides valuable structural information

when correctly interpreted.
The coupling constant (related to the size of
the splittings in the multiplet) is given the
symbol J and is measured in Hz.

J depends only on the number, type and

spatial arrangement of the bonds
separating the two nuclei, it is a property of
the molecule and is independent of the
applied magnetic field

40
Two important observations in 1H - 1H
spin-spin coupling:
(a) No inter-molecular spin-spin coupling
is observed. doesn't occur between
nuclei in different molecules.
(b) The effect of coupling falls off as the
number of bonds between the coupled
nuclei increases. Unobservable across
more than 3 intervening bonds. (large
couplings across many bonds may occur
in extended π-conjugation 41
 Spacing
in Hz

1 2
Number of 2
intervening
bonds
Coupled
nucleus
HM HA HX
Typical 1H - 1H coupling Constants
 Signal Multiplicity – The n + 1 rule
The NMR signal of a nucleus coupled to a n equivalent
hydrogens will split into a multiplet with n+1 lines.
A -CH3 group, (isolated from coupling to other protons
in the molecule) will always occur as a singlet.

A CH3-CH2- group, (isolated from coupling to other

protons in the molecule) will appear as a quartet (-
CH2-) and a triplet (CH3-) .
e.g. -1
e.g. -2

e.g. -3
e.g. -4

48
 A Spin system
A spin system is defined as a group of coupled
protons. It cannot extend beyond the bounds of
a molecule, but it may not include a whole
molecule.

ester group effectively provides a barrier (5 bonds)

against coupling between the two parts
 Rules for spectral analysis of first
• Rule 1 order spectra
• Rule 1
A group of n magnetically equivalent protons will
split a resonance of an interacting group of
protons into n+1 lines. In AnXm molecule A
protons will be split into m+1 lines. X protons will
be split into n+1 lines.

• Rule 2
The spacing (measured in Hz) of the lines in the
multiplet will be equal to the coupling constant.
• Rule 3
The true chemical shift of each group of
interacting protons lies in the center of the
(always symmetrical) multiplet.

• Rule 4

The relative intensities of the lines within

each multiplet will be in the ratio of the
binomial coefficients.
 • Rule 5
When a group of magnetically equivalent protons
interacts with another group of protons, its resonance
will take the form of a multiplet of multiplets.

In molecule AnMpXm - A protons have the

multiplicity of (p+1)(m+1)

A proton coupled to 2 different protons will be,

1) split to a doublet by coupling to the first proton

2) then each of the component of the doublet will be
split further by coupling to the second proton
This results a symmetrical multiplet with 4 lines.
54
 Spin Decoupling
it is possible to remove the splitting effects by
irradiating the sample with an additional Rf
source at the exact resonance frequency of the
proton giving rise to the splitting. (nuclei coupled
to them cannot sense them as being in either an
α or ß state for long enough to cause splitting)
The irradiated nuclei are said to be decoupled
from other nuclei in the spin system.
Decoupling simplifies the appearance of complex
multiplets by removing some of the splittings.
 Interpretation of 1H NMR spectra
Number of signals : How many different
kinds of protons
Position of signal : Environment of proton
(chemical shift)
Relative intensity of signal : Proportional
to number of protons
Splitting of signal : Number of nearby
nuclei
57
 13C NMR SPECTROSCOPY
The most abundant isotope of carbon 12
( C)
cannot be observed by NMR.
13C is a rare nucleus (1.1% natural
abundance) and its low concentration
13
coupled with the fact that C has a
relatively low resonance frequency, leads to
its relative insensitivity as an NMR-active
nucleus. ( about 1/6000 as sensitive as 'H)
(
 Coupling and decoupling in 13C NMR
spectra
13
Because the C nucleus is rare, it is
unlikely that any two adjacent C atoms
13
in a molecule will both be C.
13 13
As a consequence, C - C coupling is
13
not observed in C NMR spectra.
13
There is no splitting in C NMR
13
spectrum due to absence in C - C 13

coupling. 60
13C can be coupled strongly to protons.

1H
Therefore all nuclei are irradiated so
1
that all H are fully decoupled from the
13C nuclei.

This is termed broad band decoupling or

noise decoupling.
13
Thereby C NMR spectra usually appear
as a series of singlets.
 1H
If is not decoupled from 13C,
the
13
signals in C spectrum appear as
multiplets.

CH3 - signals appear as quartets,

-CH2 - signals appear as triplets,
-CH - groups appear as doublets
quaternary C (no H) appear as singlets.
 5 6
X-CO-O-CH2CH3

-CH2
-CH3
 SFORD : Single frequency Off- resonance
decoupling
Another method for obtaining 13C NMR spectra
(retaining the multiplicity information) involves the
use of a strong decoupling signal at a frequency
outside the range of proton resonances. This has
the effect of partially decoupling protons from the
13C nuclei.

The technique is referred as off resonance

decoupling or SFORD (Single Frequency Off
Resonance Decoupling).
 Determining 13C signal multiplicity using
DEPT
The 13C Distortionless Enhancement by
Polarisation Transfer DEPT spectrum contains
experiment is a pulsed NMR experiment which
requires a series of programmed Rf pulses to both
13
the 'H and C nuclei in a sample.
13
The resulting C DEPT spectrum contains only
signals from protonated carbons.

Non protonated carbons do not give signals in the

13C DEPT
The signals from CH3 and CH groups
(those with an odd number of attached
protons) appear oppositely phased
from those in CH2 groups (Those with
an even number of attached protons)

So signals from CH3 and CH groups

point upwards while CH2 point
downwards.
 13CNMR Spectra of Methyl
Cyclopropyl Ketone In CDCl3 Solvent at
100 MHz

69
 2 4
1
3
1 1
2
3
4

Shows 4 singlet peaks, one for each for the 4

different carbon environments in the molecule.
 2 4
1 3
2 1
3

DEPT spectrum only the 3 peaks for the

protonated carbons. CH and CH3 groups point
upwards and CH2 groups point downwards.
Carbonyl C does not appear in the spectrum as it
has no attached protons.
All resonances of protonated carbons appear as
multiplets due to coupling to attached protons.
The CH3 group appears as a quartet, the CH2 as
triplet and CH group as doublet and the carbonyl C
(with no attached protons) appears as a singlet.
Splitting shows the true value of 1JC-H.
The SFORD spectrum shows the multiplicity for all
resonances but the multiplets are narrower due to
partial decoupling of protons.
Therefore splittings are less than the true values
1
of JC-H .
80
MASS SPECTROSCOPY

- Basic Instrumentation -
This is the most accurate method for
determining the molecular mass of the
compound and it’s chemical composition.
It is possible to determine the mass/charge
ratio (m/e) of individual ions in the gas
phase.
Thereby a plot is generated for relative
abundance of each fragment against it’s
𝐦𝐚𝐬𝐬
ratio is the “Mass Spectrum”
𝐜𝐡𝐚𝐫𝐠𝐞
 IONISATION PROCESSES
The most common method of ionization
involves Electron Impact (EI) (collision of a
molecule M with an electron e) involves
electron ejection which yields an odd-
electron positively charged cation radical
+.
[M] of the same mass as the initial molecule
M.
M+e +.
[M] + 2e
The cation radical produced is known as the
+.
molecular ion ([M] )
Molecular Peak

151.16
Base peak
CH3CH2- m/Z 29
 Molecular Fragmentation
The fragmentation pattern is a
molecular fingerprint.

In addition to the molecular ion peak,

the mass spectrum consists of a
number of peaks at lower mass
number and these result from
fragmentation of the molecular ion.
90
Common Fragments and their Masses
 Isotope ratios
Fragment that contains one Br atom, the
spectrum shows 2 peaks separated by 2
79
m/e units for Br and for Br. 81

The relative intensities of the two ions are

same as the natural 79Br 81Br
abundances of both
ions are approximately 49% 51%
equal.
M M+2
fragment which contains one Cl atom, the
spectrum shows 2 peaks separated by 2
35 37
m/e units, for Cl and Cl but, the relative
intensities will be approximately 3:1.
35Cl

75% 37Cl

25%
M M+2
 CH3CH2Br79
CH3CH2Br CH3CH2Br81

m/z
108
110

Carbons and hydrogens add up to 29.

Therefore,
29 + 79 = 108
29 + 81 = 110
This compound contain Br isotope.
 CH3CH-CH3 CH3CH2Cl35
Cl
CH3CH2Cl37

m/z
78 80

Carbons and hydrogens add up to 43.

Therefore,
43 + 35 = 78
43 + 37 = 80
This compound contain Cl isotope.
Carbons and hydrogens add up to 28
28 + 35+35 = 98
28 + 37+35 = 100
28 + 35+37 = 102
This compound contain 2 Cl isotopes.
Some Common Fragments
1H NMR

13C NMR