13C NMR SPECTROSCOPY

PRESENTED BY GUIDED BY
Mr. Amit N. Chavan Prof. Dr. Ramesh Sawant
(M. Pharm. Pharmaceutics) M.Pharm., PhD
1st SEM., Roll no. 05 (Pharmaceutical
Chemistry)

Dr. V. V. P. Fs COLLEGE OF
PHARMACY, VILAD GHAT,
AHMEDNAGAR
NUCLEAR MAGNETIC RESONANCE
(NMR Spectroscopy)
A spectroscopic technique that gives us information about the
number and types of atoms in a molecule.
Nuclear magnetic resonance spectroscopy is a powerful
analytical technique used to characterize organic molecules by
identifying carbon-hydrogen frameworks within molecules.

Nuclear Magnetic Resonance

In the Involves In the

Nucleus Magnets Nucleus
 It is concerned with the magnetic properties of certain atomic
nuclei.

Involves change in the spin state at the nuclear level.

SPINNIG NUCLEUS:
proton acts as a tiny spinning bar magnet and possesses both
electrical charge and mechanical spin.
NMR is the most powerful tool available for organic structure
determination.
It is used to study a wide variety of nuclei:
1. 1H
2. 13C
3. 15N
4. 19F
5. 31P
Many atomic nuclei behave as if they spin on an axis of rotation
Nuclei are positively charged
These spinning nuclei generate tiny magnetic fields
Tiny magnets interact with an external magnetic field, denoted
B0
Proton (1H) and carbon (13C) are the most important nuclear
spins to organic chemists

Pavia, Lampman, Kriz, Vyvyan; Spectroscopy, Cengage Learning, India

edition, 2007, Pg. no. 108
Nuclear spins are oriented randomly in the absence (a) of an
external magnetic field but have a specific orientation in the
presence (b) of an external field, B0
Some nuclear spins are aligned parallel to the external field
Lower energy orientation
More likely
Some nuclear spins are aligned antiparallel to the external field
Higher energy orientation
Less likely
 PRINCIPLE OF NMR

When applied
When energy frequency is
in the form of equal to Absorption of
radiofrequency precessional energy occurs
is applied
frequency.

NMR signal is Nucleus is in

recorded. resonance
Many nuclei exhibit NMR phenomenon
All nuclei with odd number
of protons Table 1.
All nuclei with odd number The NMR Behavior of Some
of neutrons Common Nuclei
Nuclei with even numbers Magnetic Nonmagnetic
Nuclei Nuclei
of both protons and neutrons
1H 12C
do not exhibit NMR 13C 16O
phenomenon 2H 32S
15N
19F
31P
 INTRODUCTION TO 13C-NMR
Proton NMR used often for the complete elucidation of the
unknown compound.
Carbon NMR can used to determine the number of
non-equivalent carbons and to identify the types of carbon
atoms(methyl, methylene, aromatic, carbonyl.) which may
present in compound.
12C has no magnetic spin.
13C has a magnetic spin, but is only 1% of the carbon in a
sample.
The gyro magnetic ratio of 13C is one-fourth of that of 1H.
Signals are weak, getting lost in noise.
Hundreds of spectra are taken, averaged
 CHARACTERISTIC FEATURES OF
13C NMR
The area under the peak in CMR spectrum is not necessary
to be proportional to the number of carbon responsible for
the signal. Therefore not necessary to consider the area under
ratio.
Proton coupled spectra the signal for each carbon or a group
of magnetically equivalent carbon is split by proton bonded
directly to that carbon & the n+1 rule is followed.
13C nucleus is about one-fourth the frequency required to
observe proton resonance.
The chemical shift is greater for 13C atom than for proton
due to direct attachment of the electronegative atom to 13C

Gurdeep R. Chatwal, Sham K.Anand; Instrumental Methods Of Chemical

Analysis, Enlarge edition 2002,Pg. no. 2.31-2.32
 SILENT FACTS OF 13C-NMR

WHY 13C-NMR GIVES SPECTRA WHY NOT THE 12C-

NMR .

12 13
C-NMR C-NMR
no peak Gives peak

6P+6N=12 6P+7N=13
 13C CHEMICAL SHIFTS

Are measured in ppm (d) from the carbons of TMS

The correlation chart is here divided into sections
1) The saturated carbon atom which appear at Up field, nearest
to TMS(8-60ppm).
2) Effect of electronegative atom(40-80ppm)
3) Alkenes and aromatic carbon atom(100-170)
4) It contain carbonyl carbon bond. which appear at
Downfield value(155-200ppm).

Pavia, Lampman, Kriz, Vyvyan; Spectroscopy, Cengage Learning, India

edition, 2007, Pg. no. 177
 13C CHEMICAL SHIFTS ARE MOST
AFFECTED BY
Hybridization state of carbon-
Electrone gativity of groups attached to carbon-

Pavia, Lampman, Kriz, Vyvyan; Spectroscopy, Cengage Learning, India edition,

2007, Pg. no. 176-178
 13C NMR
13C signals are spread over a much wider
range
Figure #2 shows the 13C spectrum. It is much
easier to identify the compound as 1-
chloropentane by its 13C spectrum
1-Chloropentane
 PROBLEMS OF 13C

Natural abundance-
13C natural abundance is very low (1.1%).
Gyro magnetic ratio-
13C nucleus gyro magnetic ratio is much lesser than proton
nucleus. 13C-1.404; 1H-5.585.
Coupling phenomenon-
13C & 1H have I=1/2 so that coupling between them probably
occur.

Y.R.Sharma,S.Chand; Elementary Organic Spectroscopy, India edition,

2009,Pg.No.71
 PROBLEMS IN NMR CAN BE
OVERCOME BY
Fourier Transform Technique:-
Decoupling Technique:-
1) Broad Band Decoupling
2) Off Resonance Decoupling
3) DEPT (Pulse) Decoupling
Nuclear Over Hauser Phenomenon:-
 Fourier Transform NMR
Radio-frequency pulse given.
Nuclei absorb energy and process (spin) like little tops.
A complex signal is produced, then decays as the nuclei lose
energy.
Free induction decay is converted to spectrum.
Low 13C abundance
 DECOUPLING THE PROTON SPINS
1) PROTON-DECOUPLED SPECTRA
A common method used in determining a carbon-13 NMR
spectrum is to irradiate all of the hydrogen nuclei in the
molecule at the same time the carbon resonances are being
measured.
This requires a second radiofrequency (RF) source (the
decoupler) tunedto the frequency of the hydrogen nuclei, while
the primary RF source is tuned to the 13C frequency.
RF source 2
the decoupler RF source 1
continuously Pulse tuned to
Saturates Carbon-13
hydrogen's
13C signal (FID) measured
ETHYL PHENYLACETATE
2) Broadband decoupling:
In this technique, all the proton resonance can be reduced and
to get sharp CMR spectral peaks, each directly reflecting a
13C chemical shift.
The NMR spectrum of nucleus A is split by nucleus B, because
A can see B in different magnetic orientation.
3) Off resonance decoupling:
1000-2000 Hz above the spectral region.
Partially decoupled spectrum are obtained
Primary carbon nuclei- quartet
Seconadry carbon nuclei- triplet
Tertiary carbon nuclei- doublet
Qurternary carbon nuclei- single line
13C NMR SPECTROSCOPY: SIGNAL
AVERAGING AND FT-NMR

Carbon-13 NMR spectra of pentan-1-ol. Spectrum (a) is a single run, showing

background noise. Spectrum (b) is an average of 200 runs
 Characteristics of 13C NMR Spectroscopy

Factors that affect chemical shifts:

1. Chemical shift affected by nearby electronegative atoms
Carbons bonded to electronegative atoms absorb downfield
from typical alkane carbons
2. Hybridization of carbon atoms
sp3-hybridized carbons generally absorb from 0 to 90 d
sp2-hybridized carbons generally absorb from 110 to 220 d
C=O carbons absorb from 160 to 220 d
 Characteristics of 13C NMR Spectroscopy
13Cspectrum for butan-2-one
Butan-2-one contains 4 chemically nonequivalent carbon
atoms
Carbonyl carbons (C=O) are always found at the low-field end
of the spectrum from 160 to 220 d
 DEPT 13C NMR SPECTROSCOPY
Distortionless Enhancement by Polarization Transfer (DEPT-
NMR) experiment
Run in three stages
1. Ordinary broadband-decoupled spectrum
Locates chemical shifts of all carbons
2. DEPT-90
Only signals due to CH carbons appear
3. DEPT-135
CH3 and CH resonances appear positive
CH2 signals appear as negative signals (below the
baseline)
Used to determine number of hydrogens attached to each
carbon
 DEPT 13C NMR SPECTROSCOPY

Summary of signals in the three stage DEPT experiment

 13C INTERPRETATION

Count how many lines- how many types of carbons

Symmetry duplicates give same line- if there are
more carbons in your spectrum symmetry
Check chemical shift window-
Check splitting pattern-
Signal height size-
 13C INTERPRETATION

13C NMR spectroscopy provides information about:

The number of nonequivalent carbons atoms in a
molecule
The electronic environment of each carbon
How many protons are bonded to each carbon
 APPLICATION OF 13C NMR
SPECTROSCOPY
Solution structure The only method for atomic-resolution
structured etermination of bio-macro molecules in aqueous
solutions under near physiological conditions or membrane
mimeric environments.
Molecular dynamics The most powerful technique for
quantifying motional properties of biomacro molecules.
Prote in folding The most powerful tool for determining theres
idual structures of unfolded proteins and the structures of
folding intermediates.
 Ionization state The most powerful tool for determining the
chemical properties off unctional groups in bio macro
molecules, such as the ionization states of ioniz able groups at
the active sites of enzymes.
Weak inter molecular interactions Allowing weak functional
interactions between macro bio molecules (e.g., those with
dissociation constants in the micro molar to mill molar range)
to be studied, which is not possible with other technologies.
Protein hydration A power tool for the detection of interior
water and its interaction with bio macro molecules.
Hydrogen bonding A unique technique for the DIRECT
detection of hydrogen bonding interactions.
 Drug screening and design Particularly useful for identifying
drug leads and determining the conformations of the
compounds bound to enzymes, receptors, and other proteins.
Native membrane protein Solid state NMR has the potential
for determining at omicre solution structures of domains of
membrane proteinsin the irnative membrane environments,
including those with bound ligands.
Metabolite analysis A very powerful technology for metabolite
analysis.
Chemical analysis A matured technique for chemical
identification and conformational analysis of chemicals
whether synthetic or natural.
Material science A powerful tool in the research of polymer
chemistry and physics.