13C NMR Spectroscopy

Dijo Damien, Ph. D.

Assistant Professor
Research and PG Department of Chemistry
Christ College (Autonomous) Irinjalakuda
E-Mail: dijodamien@christcollegeijk.ac.in;
dijodamien1@gmail.com
 13C-NMR Spectroscopy- Why is it so insensitive?
1. Poor natural abundance 2. Low magnetogyric ratio

3. Magnetic dipole of 13C is only 1/4th as that of 1H,

hence the current produced in the receiver coil is
only 1/4th

In a nutshell overall sensitivity of 13C compared to 1H is about 1/5870

Resolution-
i. The chemical shift range in 1H NMR is only 4 ppm (320 Hz in this 80
MHz spectrum).
ii. The chemical shift range in 13C NMR 80 ppm (1600 Hz in this 20 MHz
spectrum)

Multiplicity-
i. 13C-13C coupling: not usually observed because of low natural
abundance of 13 C
ii. 13C-1H coupling: Makes the 13C spectra so complex

1H Decoupling-Noise decoupling-Broad band decoupling

i. Double irradiation using a spread of radio frequency (RF
noise) around 80 MHz to decouple entire protons -
ii. Also called-Noise decoupling-Broad band decoupling and
represented as 13C-{1H}

Is there any disadvantages for decoupling??

 Is there any disadvantages for decoupling??
➢You lost the valuable information about the multiplicity

➢ Off resonance decoupling retains the multiplicity features

Irradiation is done using a slightly off radio frequency (a few hundred Hz displaced) so that
vestigial ‘q’ remains with CH3, ‘t’ with CH2 and ‘s’ with fully substituted carbons.
➢Integration of the peak areas does not give quantitative information
1. Short interval between two successive pulses is not sufficient for saturation
➢ In the normal pulsed FT 13C work, within short Rd
Std. 13C RF pulse sequence for C nuclei with long relaxation time (T1) will not
(13C-{1H}) have fully relaxed before the next pulse is applied-
slightly saturated- low intensity
➢ If Rd is 5 x T1, this can be solved
Small symmetrical molecules (typical 13C NMR solvents including CDCl3, C6D6) have long relaxation time and end up in
the low intensity of solvent peaks

2. NOE signal enhancement for 13C nuclei connected to protons

➢ Relaxation time of 13C nuclei can be reduced very effectively, if it can transfer the excitation energy to the protons
directly attached to it.
 Nuclear Overhauser Effect
The change in the intensity of a nucleus upon irradiating a nearby nucleus

➢ NOE operates in the order CH3 > CH2 > CH > C. ie 4o C will be always of low intensity.

How much enhancement it can bring in?

Theoretical maxima = 1\2 (γirr/ γobs)
1. 1H – 1H : Max NOE= ½ (26.753/ 26.753)= 50%
2. 13C – 1H : Max NOE= ½ (26.753/ 6.728)= 198%
Does the % of NOE enhancement vary from molecule to molecule?
NOE involves relaxation - motion/tumbling of molecules - size of the molecule really matters
High molecular mass (>2000 Da) – NOE max(1H-1H) is -100%

What about viscous solvents??

For small-medium sized molecules (<1000 Da), enhancement is almost three times

For medium sized molecules (1000 - 1500 Da), no NOE

Difference Spectra
➢ To measure the NOE, we acquire (a minimum of) two spectra. The first spectrum will be irradiated at a
target resonance. The second spectrum will be irradiated at a frequency far off-resonance (-5 ppm)
and is called the reference spectrum.
➢ The reference spectrum is subtracted from the irradiated spectrum to generate the difference
spectrum. Peaks with positive intensity in the reference spectrum are interpreted as being close in
space with the irradiated peak.
How can you make use of NOE difference spectra in distinguishing the geometrical isomers?
13C Chemical shift summary

Correlation chart is divided

int 4 sections-
1. 8-60 ppm, saturated C
2. 40-80 ppm, effect of
electronegative atoms
3. 100-175 ppm, alkenes/ar
4. 155-220 ppm, carbonyl
 13C - Correlation data - Kemp 193

Lecture 16. The Importance of 13C Chemical Shifts in Structure and Stereochemistry Determination
https://www.youtube.com/watch?v=_6jFNdyZdkM&index=19&list=PLC86CC98DDF0CDDAC
Enhancing the sensitivity of insensitive nuclei-
Polarization transfer methods
Bigger population difference from a sensitive nuclei (1H) and pass it on to the
insensitive nuclei (13C, 15N), all in a predictable manner
Broadband decoupling in detail

Interconnection between spin-spin coupling and double irradiation

Coupling of two magnetic nuclei via a single bond-
Consider a C-H bond

When there is coupling between A and X

nuclei-
A → A’ (A+2E) & A’’ (A-2E)
X → X’ (X+2E) & X’’ (X-2E)
The spacing between A’ and A” as well as
X’ and X” are the same and is 4E = J

Here J ~ 15 Hz (10-13 J)
Preferred orientation of the interacting nuclei depends on the number of
intervening bonds
 Geminal
Geminal
Parallel spins are preferred
i. A absorbs RF, 1→3 & 2 →4 –
unequal energy transitions- ‘d’
ii. X absorbs RF, 1→2 & 3 →4 –
unequal energy transitions- ‘d’
Represents a system with −ve J
Electron coupled interaction alters the spin energy states of the nuclei
Vicinal
4 Situation 1-No coupling
i. A absorbs RF, 1→3 & 2 →4 –
equal energy transitions- ‘s’
ii. X absorbs RF, 1→2 & 3 →4 –
equal energy transitions- ‘s’
3 Represents a system with J=0
Vicinal
Antiparallel spins are preferred
i. A absorbs RF, 1→3 & 2 →4 –
1 unequal energy transitions- ‘d’
ii. X absorbs RF, 1→2 & 3 →4 –
unequal energy transitions- ‘d’
Represents a system with +ve J
No coupling
 Inter nuclear double resonance (INDOR) & Selective population inversion

I. Population is distorted

II. Affects the probability of other transitions

Consider the double irradiation of X1 line in the AX spectrum

Intensity of A2 increases and A1 decreases-

basis of INDOR and SPI
 Inter nuclear double resonance (INDOR)
Very low power irradiations- just to alter the
populations alone without disturbing the energy levels

Positive Irradiated and monitored lines

INDOR signal have progressive relationship

Negative Irradiated and monitored lines

INDOR signal have regressive relationship
 Selective population inversion (SPI)
An extreme case of population changes exhibited in INDOR

Let’s consider AX coupling pair- where A is 13C & X is 1H

How much enhancement SPI can bring in?

➢ Quantum theoretical treatments-
intensity follows the ratio γA and γX

➢ For 13C and 1H, ratio is 4 and max

possible intensity is (1+4)

➢ For 15N and 1H, ten fold increase in

intensity
 Multi-pulse techniques in NMR

1. APT- Attached Proton Test or J-modulated Spin-echo

to distinguish between C, CH2 & CH, CH3
2. DEPT- Distortionless Enhancement by Polarization Transfer
to identify CH, CH2 and CH3

To simplify the physics of millions of nuclear gyroscopes- we can adopt two devices
1. Nett magnetization vector (Mo)
2. Rotating frames of reference
 How the NMR signal is generated

X ω X

t
Y Y

Rotating frames of reference

Now imagine that your axis is rotating with the same angular velocity ω
X’ ω X’

t
Y’ Y’
 Refocusing spins
Spin-echo with various T2

X’ X’ X’ X’

ω- to ω+ t
ω+
t
90ox’ Y’ Y’ 180ox’ Y’ Y’
ω-
 APT- Attached Proton Test or J-modulated Spin-echo

90ox’ t t
13C ch. 180ox’
1/J 1/J CH & CH3 will give UP
CH2 and C will give DOWN
1H ch. BB Decoupling on

How does it work?

 X
Quaternary
X
Carbon
X X

t 180ox’ t
Y Y
13C 90ox’ Y Y
ch. 1/J
for 11.7
1/J
for 11.7
T, J=125 T, J=125

1H ch. BB Decoupling on
FID
signal

FT
Methine C (C-H)
X X X X

+J/2 t 180ox’ t
Y Y
13C 90ox’ Y Y
ch. 1/J
for 11.7
1/J
for 11.7
-J/2 T, J=125 T, J=125

1H ch. BB Decoupling on(no more +J/2 or –J/2)

FID
signal

FT
 Similarly methylene and methyl carbons-

How will you assign the J value in these experiments?

sp3C – 1JC-H : 125 Hz

Take an avg and take the J as 140 Hz
sp2C – 1JC-H : 160 Hz

Will this be always ok?

Eg: 1.Alkynes- 1JC-H : 250 Hz NOT Just in APT, DEPT and HMQC as well
2.CHCl3- 1JC-H : 208 Hz
 DEPT

INEPT
1/2J 1/2J 1/2J

τ = ¼ JC-H τ = ½JC-H τ= ¾ JC-H