NMR assignment

By Roy Hoffman

NMR assignment by Roy Hoffman 1

2006
Number of protons from integrals
for the unknown compound
16 Protons: 2x3, 4x2, 2x1

ppm 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0

NMR assignment by Roy Hoffman 2

2006
 Number of carbon types for the
unknown compound
11 carbon types
C11+H16O?, N?, etc.

ppm 160 140 120 100 80 60 40 20

NMR assignment by Roy Hoffman 3

2006
 Functionality from 1H chemical
shifts for the unknown compound
2x1vinyl CH
1x3 methyl alkyl
4x2 CH2

Aromatic CH, CH2

Vinyl CH2
Me-alk
Vinyl CH CH2OH Me-vinyl
ppm 9 8 7 6 5 4 3 2 1

NMR assignment by Roy Hoffman 4

2006
 Functionality from 13C chemical
shifts for
2x1vinyl CH
the unknown compound
1x3 methyl alkyl
4x2 CH2
2x1 vinyl C
1 C=O
(not CO2H because
no acidic H)
C11H16O

Vinyl C Vinyl CH2 CH, CH2

Aromatic CHOH,
C=O, CO2H CH2OH
Vinyl CH Me
ppm 175 150 125 100 75 50 25 0
NMR assignment by Roy Hoffman 5
2006
The situation so far for the unknown
compound
• C11H16O with C=O and 2 double bonds.
• This requires one ring (Without it would be C11H18O).
• There are 2 terminal methyls.
• One of the vinyls has two trans H’s
• Look for the following ring structures

NMR assignment by Roy Hoffman 6

2006
More complex structures require
2D interpretation.
•4 pairs of protons
strongly coupled to
each other &
displays 2nd order
splitting

ppm 2.15 2.10 2.05 2.00 1.95

NMR assignment by Roy Hoffman 7

2006
NMR assignment by Roy Hoffman 8
2006
 Gradient pulses
• It is possible to apply a magnetic gradient to the
sample.
• A gradient affects the signal in the following
manner. At the start of the experiment it disperses
the signal, making it disappear. Then the
application of a gradient in the opposite direction
allows the signal to be seen again.
• In combination with rf pulses that act as quantum
filters it is possible to observe correlations
between nuclei.
NMR assignment by Roy Hoffman 9
2006
Example pulse sequence

NMR assignment by Roy Hoffman 10

2006
2D Fourier transform

NMR assignment by Roy Hoffman 11

2006
 Types of 2D
• Through bond
– COSY, TOCSY, heteronuclear correlation
(HSQC, HMBC, HMQC), 2D-INADEQUATE
• Through space
– NOESY, ROESY, HOESY
• Resolving the Hamiltonian
– J-resolved, MQ-MAS

NMR assignment by Roy Hoffman 12

2006
 COSY

Basic pulse sequence for COSY

Relaxation time Evolution time, t1 Acquisition of fid, t2

90°±x 90°x

NMR assignment by Roy Hoffman 13

2006
COSY of ethylbenzene

NMR assignment by Roy Hoffman 14

2006
Artifacts in COSY

NMR assignment by Roy Hoffman 15

2006
 COSY of the unknown compound
Two networks: 1

CH3CH2CH=CHCH2
2

and C=O 3

Strong signals are 2

4

& 3 bond, weak

signals are 4 bond 5

ppm

ppm 5 4 3 2 1

NMR assignment by Roy Hoffman 16

2006
 COSY example:12,14-
ditbutylbenzo[g]chrysene

NMR assignment by Roy Hoffman 17

2006
Division into coupling networks

NMR assignment by Roy Hoffman 18

2006
Nuclear Overhauser effect (NOE)
• When a specific nucleus is magnetically excited
and its neighbor is at equilibrium, relaxation
occurs between the two nuclei.
• The smaller the physical distance between the
equilibrium nucleus and the excited nucleus, the
bigger the expected change.
• This dependence can be used to estimate the
distance between nuclei.
• NOE is used to observe through space correlations
in NOESY and ROESY experiments.

NMR assignment by Roy Hoffman 19

2006
 NOESY uses NOE to correlate
through space
Physically close so strong NOE
but 6-bonds away so no COSY

NOESY COSY

NMR assignment by Roy Hoffman 20

2006
 Basic pulse sequence for NOESY

90°x 90°x 90°±x

Relaxation time Evolution Mixing Acquisition of
time, t1 time, tm fid, t2

NMR assignment by Roy Hoffman 21

2006
Aromaric region of the 2D NOESY spectrum of 12,14-
ditbutylbenzo[g]chrysene showing connectivity and
separation into four color-coded proton groups

NMR assignment by Roy Hoffman 22

2006
 NOESY of the unknown compound
1H NMR is assigned Me7 1
4 9
3 5 82
Correlation between
the two networks 1

A ring is required so
close with C=O needed 4

to fit 13C spectrum

O

10 5 3 1 5

9 6 4 2 ppm

8 7 ppm 5 4
NMR assignment by Roy Hoffman 3 2 1
23
2006
One-bond 1H-13C correlation
(HSQCSI) of butan-2-ol
H1
H4
H2 OH
H3

C4 10

20
C1

30
C3

40

50

60

C2
ppm
ppm 3.5 3.0 2.5 2.0 1.5 1.0

NMR assignment by Roy Hoffman 24

2006
One-bond 1H-13C correlation
(HSQCSI) of ethylbenzene

NMR assignment by Roy Hoffman 25

2006
 Basic pulse sequence for heteronuclaer
correlation
Observation
channel 1H Relaxation Evolution
time time, t1

Coupled 90°x Acquisition

of fid, t2
channel 13C

90°±x

n = 3.98

NMR assignment by Roy Hoffman 26

2006
 HSQC of the unknown compound
Me7 1
4 9
3 5 8 2

Me75 12 20

89
40

Assign carbons
60

attached to protons
80

100

120

4
3 ppm

ppm 5 4 3 2 1

NMR assignment by Roy Hoffman 27

2006
 Multi-bond 1H-13C correlation
(HMBC) of 2,4-dichlorophenol
H3 H6
H5
Correlation table
C6 C\H 3 5 6
C2 120
6
C4 125
C5
C3 OH
130
2 3
1
2
Cl
135 4 3
6

5 3
140 5 3
4
145 3 3
Cl
C1 150
ppm 1 3 3
ppm 7.4 7.3 7.2 7.1 7.0 6.9

NMR assignment by Roy Hoffman 28

2006
Multi-bond 1H-13C correlation
(HMBC) of ethylbenzene
Correlation table
C\H 1' 2' 2 3 4
1' 2
2' 2 1
1 2 3 3
2 3 3 3
3 3
4 3
NMR assignment by Roy Hoffman 29
2006
NMR assignment by Roy Hoffman 30
2006
 HMBC to assign other carbons of
the unknown compound
Me7 1
4 9
3 5 8 2
•3-bond
correlations 25

8 1-bond
strongest 50

correlation
75

O
100

10 5 3 1
9 6 4 2 125

8 7 6 150

7
10 175

ppm

ppm 5 4 3 2 1

NMR assignment by Roy Hoffman 31

2006
 Use correlation table
H1 H2 H3 H4 H5 Me7 H8 H9
C1 2
C2 2 3
O C3 3 2 3

3 C4 3 2
10 5 1
9 C5 3
6 4 2 C6 2 3 3
8 7
C7 3 3
Me7
C8 3 2
C9 2
C10 3 4 3

NMR assignment by Roy Hoffman 32

2006
Assignment example camphor
8 9
7 10

1 O
6 2

4
5 3
NMR assignment by Roy Hoffman 33
2006
8 9
1H NMR of camphor
7 10

1 O

6 2

4
5 3

ppm 2.2 2.0 1.8 1.6 1.4 1.2 1.0

NMR assignment by Roy Hoffman 34

2006
 13C-NMR of camphor
8 9
7 10

1 O

6 2

4
5 3

ppm 200 175 150 125 100 75 50 25

NMR assignment by Roy Hoffman 35
2006
NOESY of camphor

1.0

1.5

2.0

ppm
ppm 2.0 1.5 1.0
NMR assignment by Roy Hoffman 36
2006
NOESY of camphor
0.8

0.9

1.0
ppm

ppm 2.2 2.0 1.8 1.6 1.4

NMR assignment by Roy Hoffman 37
2006
 NOESY of camphor
4 5e 6e
3e

0.8

9 8 9
7 10

1 O
0.9
6 2
10
4
5 3
8

1.0
ppm

ppm 2.2 2.0 1.8 1.6 1.4

NMR assignment by Roy Hoffman 38
2006
 COSY of camphor
Red lines show artifact regions
9
108
1.0
8 9
7 10
5a
6a 1.5 1 O

6e 6 2
3a 4
5e 2.0 5 3
4
3e
ppm
ppm 2.0 1.5 1.0
NMR assignment by Roy Hoffman 39
2006
One-bond 1H-13C correlation HSQC
10
of camphor8 9
3e 4 5e 3a 6e 6a5a
10 10
8 9
15 7 10
8
9 20 1 O

25 6 2
5
6 30 4
5 3
35

40
4
3 ppm
ppm 2.2 2.0 1.8 1.6 1.4 1.2 1.0
NMR assignment by Roy Hoffman 40
2006
 8 9
7 10

Multi-bond correlation 1 O

6 2
HMBC of camphor
4
5 3
H3 H4 H5 H6 H8 H9 H10
C1 3 2 3 3 2
C2
C3
C4 2 3 3
C5 3
C6 3
C7 2 2 2 3
C8 1 3
C9 3 1
C10 1
NMR assignment by Roy Hoffman 41
2006
 Multi-bond correlation HMBC of
camphor 10
8 9
3e 4 5e 3a 6e 6a5a

50
8 9
7 10

1 100
O

6 2
150
4
5 3

200

2 ppm
ppm 2.0 1.5 1.0
NMR assignment by Roy Hoffman 42
2006
Multi-bond correlation HMBC of
camphor 10
8 9
3e 4 5e 3a 6e 6a5a
10 10

8 8 9
9 20 7 10
5
6 30 1 O

6 2
4 40
31 4
50 5 3

7 60

ppm
ppm 2.0 1.5 1.0
NMR assignment by Roy Hoffman 43
2006
 Methyls
1H-NMR of carvone
7

O
6 1
2

5 3
4
9 aliphatic
10 10
8
9
CH, CH2

ppm 6 5 4 3 2

NMR assignment by Roy Hoffman 44

2006
 13C-NMR of carvone
7

O
6 9
6 1 Methyls
2

5 3
4 Aliphatic
Vinyl CH, CH2
8
10 9
Solvent
2

ppm 180 160 140 120 100 80 60 40 20

NMR assignment by Roy Hoffman 45
2006
 COSY of carvone 10
7
5 7

2
O
6 1
2

3 5 3
4

4
8
10 9

9 5

6 ppm
ppm 6 5 4 3 2
NMR assignment by Roy Hoffman 46
2006
COSY of carvone
4 3A 5A 5B 3B
7

O
6 1
2
2.2
5 3
4

2.4
8
10 9

2.6

ppm
ppm 2.7 2.6 2.5 2.4 2.3 2.2 2.1

NMR assignment by Roy Hoffman 47

2006
 One-bond 1H-13C correlation HSQC
of carvone 3A3B
5B
7 10
6 9 4 5A 7
7
10 20 O
5 6 1
4 2
40
3 5 3
4
60

80 8
10 9

100

9
120

140
6 ppm
ppm 6 5 4 3 2
NMR assignment by Roy Hoffman 48
2006
 Multi-bond 1H-13C correlation
7
HMBC of carvone 10
9 3B 7 6 1
O
6 4 3A5B
5A 2
7
10 25
5
4
3
5
34 50
8
10 9
75

100
9
125
1
6
8 150

175

2 ppm
ppm 6 5 4 3 2
NMR assignment by Roy Hoffman 49
2006
 Multi-bond correlation HMBC of
carvone
H3 H4 H5 H6 H7 H9 H10
C1 2
C2 2 3 3
C3 7 3
C4 3 3
O
C5 3 6 1
C6 2

C7 5 3 1
4
C8 2
C9 8 1 3
10 9
C10 3 1

NMR assignment by Roy Hoffman 50

2006