Theory and application of 2D NMR

spectroscopy in structure assignment

to organic compounds

Dr D. I. Brahmbhatt
Professor of Organic Chemistry
Department of chemistry
Sardar Patel University
Vallabh Vidyanagar – 388 120
Gujarat
Email: drdibrahmbhatt@gmail.com
Mobile: 9427547665
DQF 1H-1H COSY : It is Double Quantum Filtered 1H-1H Correlation
Spectroscopy. The “off-diagonal or cross peaks” in it
correlates the coupled protons.
HETCOR : It is 13C-1H Correlation Spectroscopy. It correlates 13C nuclei
with directly attached (i.e. coupled) protons.

HMQC : It is Inverse Detected version of 1H-13C COSY.

HMBC : It is Proton Detected, Long range 1H-13C COSY.

INADEQUATE : It is 13C-13C COSY. It correlates directly attached 13C-13C

coupling.
TOCSY : It is Totally Correlated Spectroscopy. It enables us to see
correlation among nuclei that are not directly coupled but
within the same spin system.
 Correlation Spectroscopy(2D NMR)
• HOMO correlation spectroscopy

• 1H-1H correlation spectroscopy

• COrrelation SpectroscopY – COSY

• 1H-1H COSY

• HETero CORrelation spectroscopy – HETCOR

• 1H-13C HETCOR

• DQF-COSY – Double Quantum Filtered COSY

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt
• HMQC – Heteronuclear Multiple Quantum Coherence

• HMBC – Heteronuclear Multiple Bond Coherence

• NOESY – Nuclear Overhouser Enhancement SpectroscopY

• INADEQUATE – Incredible Natural Abundance DoublE

QUAntum Transfer Experiment

• TOCSY – TOtally Correlated SpectroscopY

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Principle of Fourier Transform

• Fourier – a French mathematician gave equation to solve combined

waves.
• Two combined waves – four answers
2 – frequencies
2 – amplitudes
• Five combined waves – ten answers
5 – frequencies
5 - amplitudes

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

• FID – Free Induction Decay.
• Time domain spectrum
• FID FT Frequency Domain Spectrum.

• Length of FID – more sharp signal

• Length of FID short - broad signal
Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt
Fourier transform of FID

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

FID spectrum of Methanol and Ethanol

CH3
49.0 CH2 CH3
57.0 17.0

Methanol Ethanol

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt
2D NMR spectrum of chloroform

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

2D NMR spectrum of β-chloro acrylic acid

Cl C C COOH
H H

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

heptan-3-one
1H - 1H COSY spectrum of 3-heptanone

1 3 5 7
2 4 6

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

1H - 1H COSY spectrum of m-dinitro benzene

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Terpenoids, Ipsenol and Caryophyllene oxide :

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

300 MHz PMR spectrum of Ipsenol

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

t1 (evolution time) is order of micro to millisecond while t2 (acquisition time) is
order of seconds.

During t1, spins, which are mutually coupled, precess under influence of both
nuclei’s chemical shifts and thus give rise to peaks in which u1 does not equal
u2.

In the general case, 1H-1H COSY spectra are interpreted as giving rise to off-
diagonal or cross peaks or contours for all protons that have significant (i.e.
measurable J-J) coupling.

Simply, the cross peaks correlates coupled protons.

1H-1H COSY of Ipsenol:

F2 is found on the bottom (X-axis), with proton scale as usual. It generates
from t2.
F1 is displayed on the right (Y-axis), with proton scale running from top to
bottom. It generates from t1.

A proton spectrum is displayed opposite to F1 and F2, and are given for
convenience.

These 1-D spectra are not part of the 1H-1H COSY spectrum in reality.
From the upper right to the lower left runs the “diagonal”, a series of
absorptions in which u1 equals u2.

These diagonal cross peaks provide nothing in the way of useful information
beyond the simple 1-D 1H spectrum.

On the either side of the diagonal and symmetrically disposed are the cross
peaks.

The symmetry in this type of spectrum is oftentimes imperfect.

We can make one further refinement before undertaking detailed

interpretation, that decreases the “clutter” along the diagonal.
1H - 1H COSY spectrum of Ipsenol

Isolated CH2
CH2

5 6 7
H
4 CH2
HO 3 8

H3C 2 CH3
1

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

By simply adding a third π/2 pulse immediately following the second π/2
pulse in our simple COSY sequence and changing nothing else, we have the
pulse sequence for the very popular double-quantum filtered 1H-1H COSY (DQF-
COSY) experiment.

The purpose of the third pulse is to remove or “filter” single quantum

transitions so that only double quantum or higher transitions remain.

In practical terms, the double quantum filter will select for systems with at
least two spins (minimum AB or AX).

Thus methyl singlets (noncoupled) will be greatly reduced.

Higher quantum filtering is possible but it is generally not used, e.g. triple-
quantum filter.

By using 1H-1H DQF-COSY, the spectrum seems “cleaner” especially along the
diagonal making the task of interpretation significantly easier.

Mostly all COSYs are recorded as DQF-COSYs due to improved appearance.

1H - 1H DQF -COSY spectrum of Ipsenol
Isolated
7 8 4 5 2 & OH
1
1 3
3
2 & OH
CH2
5
5 6 7
H 4
4 CH2
HO 3 8

Isolated H3C 2 CH3

1
8

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Sr. HETCOR
1 HETero-nuclear CORrelation
spectroscopy
2 It is 13C-1H Correlation
Spectroscopy.
3 It is Carbon detected
4 It correlates 13C nuclei with
directly attached (i.e. coupled)
protons
5 It shows 1JCH
6 Sensitivity less due to low value Sensitivity high due to high
of γ for 13C
7 On X-axis (F2) there is Carbon
scale while on Y-axis (F1) there scale while on Y-axis (F1) there
is proton scale
8 There will be no diagonal and
no symmetry . It will be the
case when F1 and F2 represent
different nuclei
9 It is simpler than HMBC. It
shows one bond correlations
while two and three bond
correlations are absent
HMQC
Hetero-nuclear Multiple
Quantum Coherence
It is Inverse Detected version of
13C-1H COSY.
It is proton detected
It correlates 13C nuclei with
directly attached (i.e. coupled)
protons
It shows 1JCH
value of γ for 1H. It is 30 folds
then normal HETCOR
On X-axis (F2) there is proton
is Carbon scale
There will be no diagonal and
no symmetry .
It is simpler than HMBC. It
shows one bond correlations
while two and three bond
correlations are absent
HMBC
Hetero-nuclear Multiple Bond
Coherence
It is Long range 1H-13C COSY.
It is Proton Detected
It correlates 13C nuclei with protons
which are not directly attached (i.e.
coupled) but separated by 2 or 3
bonds
It shows Mainly 2JCH and 3JCH
Sensitivity high due to high value of γ
for 1H. It is 30 folds then normal
HETCOR
On X-axis (F2) there is proton scale
while on Y-axis (F1) there is Carbon
scale
There will be no diagonal and no
symmetry .
It is complex than HMQC and
HETCOR . It shows two and three
bond correlations while one bond
correlations are absent
HETCOR spectrum of Ipsenol
Isolated
C7 CH2 C8 C4 C3 C5 C2
C6
C1

Isolated CH2
CH2

5 6 7
H 4
CH2
HO 3 8

H3C 2 CH3
1

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

HMQC (Inverse detected HETCOR) spectrum of
Caryophyllene oxide

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

HMBC spectrum of Ipsenol

Isolated CH2

CH2

5 6 7
H 4
CH2
HO 3 8

H3C 2 CH3
1

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

butan-2-ol
butan-2-ol
HETCOR spectrum of 2-butanol

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

NOESY Spectrum of Pamoic Acid

CH2

H7
H6
H8
H5

H6 H7 H1
H1 H5 H8 CH2

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

INADEQUATE Spectrum of Caryophyllene oxide
C13 C9 C7
C8

15 2 3
12
H3C H CH3
1 4
H3C 11
5
14
9
O
10 6
8
H
7
H2C
13

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

 C9 C3 C6+C7 C14
C5 C4 C1 C10 C11 C15 C2 C12

15 2 3
12
H3C H CH3
1 4
H3C 11
5
14
9
O
10 6
8
H
7
H2C
13

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt
COSY spectrum of butanoic acid
c b a

a c
COOH
b

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

HETCOR spectrum of butanoic acid
c b a

a c
COOH
b

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

COSY spectrum of p-amino-ethylbenzoate

a b
CH3CH2O O

e e

d d

NH2
c

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

HETCOR spectrum of p-amino-ethylbenzoate
5 3
6 2 1
7 4

a b
CH3CH2O O
1 2 7
4
e e 5

d d 3
6
NH2
c

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

1-chloropropan-2-ol
1-chloropropan-2-ol
COSY spectrum of 1-chloro-2-propanol

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

HETCOR spectrum of 1-chloro-2-propanol

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt

Department of Chemistry, SPU, VVN Dr. D. I. Brahmbhatt