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NMR Spectroscopy

2.1. Introduction

NMR: Map of Carbons with Hydrogens

 NMR is the most valuable

 Used to determine relative location of atoms

 Maps carbon-hydrogen framework of molecules

 Depends on very strong magnetic fields

 More advanced NMR techniques are used in biological chemistry to

study protein structure and folding

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First the 1-D techniques:

Experiment Nuclei Types of information, applications

J-modulated spin-echo 13C CH and CH3 carbon nuclei give positive signals, 4 and
(attached proton test, CH2 carbons give negative signals. An aid to
APT) assignment.

INEPT 1H – 13C The INEPT pulse sequence is used as a component of

many 2D experiments e.g. HSQC

DEPT 13C Tells how many hydrogen atoms are directly bonded to
a carbon nucleus: CH, CH2, CH3. Disadvantage: no
signals from 4 carbon atoms.

Selective TOCSY 1H Allows identification of all the protons belonging to a

common coupled spin system.

1D-INADEQUATE 13C Exact 13C-13C coupling constants without the need to

synthesise 13C enriched compounds.

Second the 2-D techniques:

Experiment Nuclei Types of information, applications
Heteronuclear J-resolved 13C 13C-1Hcoupling constants, number of directly
13C NMR spectroscopy bonded protons (as in DEPT).
Homonuclear J-resolved 1H 1H Useful in determining chemical shift values in
NMR spectroscopy complicated spectra, identifying peaks of
HOMO2DJ multiplets.
1H-1H COSY 1H Assigning signals in complicated spectra.
Long-range COSY 1H Assigning signals of protons separated by four or
more bonds where the couplings are small.
1H-13CCOSY (HMQC, 1H 13C Assigning signals in proton and carbon spectra,
Heteronuclear Multiple starting from known signals.
Quantum Coherence)
HMBC (Heteronuclear 1H 13C Assigning 1H and 13C signals on the basis of long
Multiple Bond Correlation) range couplings.
2D-TOCSY 1H Allows identification of all the protons belongng to
a common coupled spin system.
NOESY 1H Gives evidence for spatial proximity of nuclei using
nuclear Overhauser effect.
EXSY 1H Qualitative evidence of exchange processes.
2D-INADEQUATE 13C Assigning signals by detecting couplings between
adjacent 13C nuclei.
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2.2. Principle

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2.4. Sample Preparation

 5–40 mg typical → only μg needed for 1H NMR

 solvents: typically deuterated to avoid overlap with sample peaks
 CCl4, CDCl3, C6D6, acetone-d6, DMSO-d6, CD2Cl2, CD3CN, etc.
 CDCl3 has become the ―standard‖ solvent
 single peak observed, relatively far away from other peaks (δ 7.27 ppm)
 best solvents are those that are volatile and not very hygroscopic
 standard: varies with nucleus of interest for 1H and 13C: SiMe4 (TMS) = 0 ppm

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2.5. The NMR Spectrum

Effect of Field Strength on NMR Spectrum

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The Nature of NMR Absorptions

 1HNMR of methyl acetate has 2 equivalent kinds of H’s so shows 2 peaks
 Electrons in neighboring bonds shield or expose nuclei from magnetic
field
H’s on C next to electron withdrawing C=O

H’s on C next to electron withdrawing O

 Intensity of 1H NMR peak is proportional to # of equivalent H’s

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 13C NMR of methyl acetate has 3 kinds of C’s so shows 3 peaks

 Electrons in neighboring bonds shield nuclei from magnetic field
C next door to electron withdrawing C=O
C next to electron withdrawing O

C of C=O

 Intensity of 13C NMR peak is not related to # of equivalent C’s

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Example:
 How many signals would you expect each to
have in its 1H and 13C spectra?
1H 13C

CH3 CH3
C C
CH3 CH3

CH3
CH3

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Solution:
 How many signals would you expect each to
have in its 1H and 13C spectra?
1H 13C

CH3 CH3 1 CH3 CH3 2 CH3 CH3

C C C C C C
CH3 CH3 CH3 CH3 CH3 CH3

CH3 H H CH3
CH3
3 H CH3 5 C
C
C CH3
H CH3

O H C C
O O C
H
H H

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13C NMR Example: 2-butanone

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13C NMR Example: p-bromoacetopheone

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Learning Check:
Assign resonances in the given 13C spectrum of methyl propanoate

O
1 3 2 4
CH3 O C CH2 CH3

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Solution:
Assign resonances in the given 13C spectrum of methyl propanoate

O
1 3 2 4
CH3 O C CH2 CH3

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13.7 Uses of13C NMR: Example

 Evidence for product of elimination of 1-chloro-methyl cyclohexane
Cl
CH3 CH2 CH3
KOH
or
ethanol

Expect 5 different C’s; Expect 7 different C’s;

3 sp3 resonances  20-50 5 sp3 resonances  20-50
2 sp2 resonances  100-150 2 sp2 resonances  100-150

CH3

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Nonequivalent H’s
 If replacement of each H with ―X‖ gives a different constitutional
isomer then the H’s are in constitutionally heterotopic
environments and will have different chemical shifts
 – they are nonequivalent under all circumstances

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Equivalent H’s
 Two H’s that are in identical environments (homotopic) have the
same NMR signal
 Test by replacing each with X if they give the identical result, they
are equivalent (homotopic)

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Enantiotopic H’s
 If H’s are in environments that are mirror images of each other, they
are enantiotopic
 Replacement of each H with X produces a set of enantiomers
 The H’s have the same NMR signal (in the absence of chiral
materials)

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Diastereotopic H’s
 In a chiral molecule, paired hydrogens can have different
environments and different shifts
 Replacement of a pro-R hydrogen with X gives a different
diastereomer than replacement of the pro-S hydrogen
 Diastereotopic hydrogens are distinct chemically and
spectrocopically

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Learning Check:
 Identify sets of H’s as
Unrelated (U), homotopic (H), enantiotopic (E), or diasterotopic (D)

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Solution:
 Identify sets of H’s as
Unrelated (U), homotopic (H), enantiotopic (E), or diasterotopic (D)

E D D

D D H

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2.6. Chemical Shifts (δ)

 is the resonant frequency of a nucleus relative to a standard in a magnetic

field. Often the position and number of chemical shifts are diagnostic of the
structure of a molecule.
 Tetramethylsilane, (CH3)4Si, (TMS) is the usual reference compound chosen
for both 1H and l3C NMR due to the following advantages as a reference
compound:
 Relatively inert, has low b.p. (26.5°C) and can be easily removed after use;
 Gives a sharp single signal in 1H and l3C due to equivalent H and C atoms;
 The presence of Si in the TMS makes its signal occurs outside the normal
range observed for organic compounds;
 the chemical shift of TMS is not substantially affected by complexation or
solvent effects because the molecule doesn't contain any polar groups.
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Chemical shifts can be measured in Hz but are more usually expressed in ppm.

 Deshielded (downfield)—a nucleus whose chemical shift is increased, feels

stronger magnetic field due to the removal of electron density, magnetic
induction, etc in neighboring atoms or groups
 Shielded (upfield)—a nucleus whose chemical shift is decreased, feels
weaker magnetic field due to the addition of electron density, magnetic
induction, etc in neighboring atoms or groups

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2.7. Proton (1H) chemical shift

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Chemical Shifts in 1H NMR

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2.8. Factors affecting 1H chemical shift

1) Electronegativity Effects

 Nuclei tend to be deshielded by which electron-withdrawing groups (EWG)

(higher EN atom, X, O, N).
 EWG (-OH, -OCOR, -OR, -NO2, halogen) attached to an aliphatic carbon chain
cause a downfield shift of 2-4 ppm when present at Cα and have less than half
of this effect when present at C

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2) Hybridization Effects
IH chemical shifts are sensitive to the hybridization of the atom to which the IH
nucleus is attached (sp3, sp2 etc.). Generally IH chemical shifts

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3) Resonance Effects

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4) Hydrogen Bonding
The chemical shift depends on how much
hydrogen bonding is taking place
R
(observed in high concentrated solutions).
O H H
 Hydrogen bonding lengthens the O-H
O H O R bond and reduces the valence electron
density around the proton
R  it is deshielded and shifted downfield in
the NMR spectrum.

Alcohols vary in chemical shift D2O-exchangeable (peak for OH proton

from 0.5 ppm (free OH) to about in alcohol and NH in amines disappears
5.0 ppm (lots of H bonding). upon shaking with D2O) 36

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 Resonance, electronegativity of oxygen and the formation of hydrogen bonding

withdraw electron cloud from the acid protons.
 Thus, protons attached to carboxylic acids are the least shielded protons and
have a chemical shift of 10-12 ppm.

H3C O
O In methyl salicylate, which has strong
H
internal hydrogen bonding, the NMR
O absorption for O-H is at about 14 ppm,
(highly downfield)

Notice that a stable 6-membered ring is formed 37

δ- δ+ δ-
-I Carboxylic acids have strong
hydrogen bonding - they
form dimers.

δ- δ+ δ-

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5) Magnetic Anisotropy
 The chemical shift of a nucleus may also be affected by the presence in its
vicinity of a magnetically anisotropic group (e.g. an aromatic ring or carbonyl
group).
 In an aromatic ring, the "circulation" of electrons effectively forms a current
loop which gives rise to an induced magnetic field

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Aromatic protons
 = 7-8 ppm

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Vinyl (Olefinic) protons,

 = 5-6 ppm

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Acetylene protons
̃ ≈ 2.5 ppm

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Aldehyde proton
= 9-10 ppm

Electronegative
oxygen atom

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Values for Coupling Constants

Chapter 13 44
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2.9. Carbon (13C) chemical shift

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2.10. Factors affecting 13C chemical shift

1) Hybridization Effects

 Saturated sp3 carbons are found between 50 and 80 ppm,

 Unsaturated carbons (sp2 or sp hybridized) between 80 and 160 ppm,
including aromatic carbons
 carbonyl carbons from ~160 to 220 ppm.

2) Electronegativity effect

 electron withdrawing substituents, heteroatoms and alkyl groups α(-C-X)

or (-C-C-X) to a carbon atom cause downfield shifts of the 13C

resonance.
 Substituents γ(-C-C-C-X) cause upfield shifts attributed to steric interaction.

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Electronegativity of X Substituent Carbon

α β γ
X CH CH2 CH2
2.1 H 0
2 0 0
2.5 CH +9 +10 -2
2.5 SH
3 +11 +12 -6
3.0 NH2 +29 +11 -5
3.0 Cl +31 +11 -4
4.0 F +68 +9 -4

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2.11. Differences Between 1H and 13C NMR

 range of 13C chemical shifts is very large compared to 1H

 13C chemical shifts are more sensitive to small changes in chemical
environment
 Influence of electronegative atoms on chemical shift greater in 13C NMR than
1H NMR

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2.12. Integration

 The area under a particular peak in a nuclear magnetic resonance spectrum is

proportional to the number of protons in the molecule producing the peak.

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2.14. Spin - Spin Coupling

 A magnetic nucleus can sense the presence of other nuclei through the bonds of
the molecule the signals will exhibit fine structure (splitting or multiplicity).
 Multiplicity arises because if an observed nucleus can sense the presence of
other nuclei with magnetic moments.
 The coupling constant (J) (related to the size of the splittings in the multiplet)
given the symbol and is measured in Hz. It is represents the number of
intervening bonds between the coupled nuclei.
 Signal Multiplicity - the n+l rule.
 Spin-spin coupling gives rise to multiplet splittings in IH NMR spectra.
 The NMR signal of a nucleus coupled to n equivalent hydrogens will be split into
a multiplet with (n+l) lines.
 The relative intensity of the lines in multiplet will be given by the binomial
coefficients of order ―n‖.
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2.15. Writing Up Your Experimental Assignments

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Spin-Spin Splitting Example:

Integration shows ratio Triplet ~1.2
Quartet ~2.8
(~deshielded) (shielded)
2 H’s see 3 3 H’s see 2
Shift ~7.8 & 6.8 Singlet at 3.8
neighbors neighbors
(deshielded) (deshielded) (typical (typical CH3-CH2)
2 H’s see 1 neighbor 3 H’s see 0 CH3-CH2-c=o)

2x (1+1=2 doublet) neighbors

• (typical para pattern) (typical CH3-O) 3

a b 2

3
a b
2
a
2 b

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Learning Check:
From the 1H NMR of C4H10O propose a structure.

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Solution:
From the 1H NMR of C4H10O propose a structure.

CH3 CH2 O CH2 CH3

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More Complex Spin-Spin Splitting

Patterns: trans-cinnamaldehyde
 Spectra more complex if overlapping signals, multiple
nonequivalence
Doublet of
Doublets (dd)
Doublet ~9.8
~6.7
(deshielded)
(~deshielded)
1 H see 1 1 H see’s 2
neighbor different
•(typical aldehyde)
1 neighbors
3
b (typical
CH-CH-CH)
3 a c

1
a
1 b
2
2 c a
1 b

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More Complex Spin-Spin Splitting

Patterns: trans-cinnamaldehyde
 Spectra more complex if overlapping signals, multiple
nonequivalence Doublet at 7.5 a=d
(deshielded) Doublet of
2 H’s see 1
1 H see’s 1 neighbor Doublets (dd)
Doublet ~9.8 neighbor (typical CH-CH) ~6.7
(deshielded) (large J value (~deshielded)
1 H see 1 typical trans alkene H) b = dd 1 H see’s 2
neighbor 2 H’s see 2 different
•(typical aldehyde)
1 different neighbors
3
b neighbors (typical
CH-CH-CH)
3 a c (typical CH-CH-CH)

1
a
1 b
2
2 c a
1 b

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Uses of 1H NMR Spectroscopy

 Determine the regiochemistry of hydroboration/oxidation of
methylenecyclohexane. Which structure gives this 1H NMR?

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Which of the following nuclei does not show

magnetic behavior?
20% 20% 20% 20% 20%
1. 1H

2. 2H

3. 12C

4. 13C

5. 17O

1 2 3 4 5

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Which of the following is not true of 13C NMR

spectra?
1. The number of carbon atoms 20% 20% 20% 20% 20%
in a molecule can be
ascertained.
2. The number of hydrogen
atoms in a molecule can be
ascertained.
3. Certain functional groups can
be deduced from the
locations of the peaks.
4. Both the number of carbon
atoms and the number of
hydrogen atoms in a
molecule can be ascertained.
5. All of these.
1 2 3 4 5

Chemically equivalent nuclei always show a

single absorption.
50% 50%
1. True
2. False

1 2

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How many signals will appear in the 13C NMR

spectrum of the following molecule?
20% 20% 20% 20% 20%

1. 1
2. 2
3. 3
4. 4
5. 5

1 2 3 4 5

How many signals will appear in the 13C NMR

spectrum of the following molecule?
OCH3 20% 20% 20% 20% 20%

1. 3
2. 4
3. 5
4. 6
5. 7
1 2 3 4 5

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Which of the following molecules best fits the

following 13C NMR data?
13C NMR data: 20, 22, 32, 44, 67 ppm

25% 25% 25% 25%

1.
CH3 2.
HO CH3
HO

H3C
H3C

3. 4.
HO HO

H3C CH3
H3C
CH3

1 2 3 4

What is the relationship between Ha and Hb in

the following compound?
Cl 20% 20% 20% 20% 20%

Ha
Hb

1. chemically unrelated
2. homotopic
3. enantiotopic
4. diastereotopic
5. none of these
1 2 3 4 5

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What is the relationship between Ha and Hb in

the following compound?
20% 20% 20% 20% 20%
OH
Hc
Ha Hb

1. chemically unrelated
2. homotopic
3. enantiotopic
4. diastereotopic
5. none of these
1 2 3 4 5

Order the following protons from lowest to

highest chemical shift value.
20% 20% 20% 20% 20%
O
Hb Hd
Hc
Ha

1. Ha < Hc < Hb < Hd

2. Ha < Hc < Hd < Hb
3. Hc < Ha < Hd < Hb
4. Hc < Ha < Hb < Hd
5. Hc < Hd < Ha < Hb

1 2 3 4 5

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1H NMR will allow one to distinguish between

the following two molecules:

H Br Br H 50% 50%

1. True
2. False

1 2

The painkiller Demerol has the structure shown below. How

many peaks would you expect to see in the 13C NMR spectrum of
this substance?

CH3 20% 20% 20% 20% 20%

N

O
O

1. 10
2. 11
3. 12
4. 14
5. 15 1 2 3 4 5

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The spectrum shown could represent the

molecule in the illustration.

50% 50%

1. True
2. False 1 2

Which of the following molecules best fits the

following NMR spectrum?

20% 20% 20% 20% 20%

1. 2.

3. 4. 5.
1 2 3 4 5

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Which of the following molecules best fits the

following NMR spectrum?

20% 20% 20% 20% 20%

1. 2.
OCH3
OCH3

3.
O
4. 5.
O O

1 2 3 4 5

Which of the following molecules best fits the

following NMR spectrum?

20% 20% 20% 20% 20%

1. 2. 3.
O OH 4. 5.
OH HO HO

O
HO HO
OH OH 1 2 3 4 5

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Which of the following molecules best fits the

following NMR spectrum?

20% 20% 20% 20% 20%

1. 2.
Cl OH

3. 5.
4. NH2
NH2
1 2 3 4 5

Which of the following molecules best fits the

following NMR spectrum?

20% 20% 20% 20% 20%

1. 2.
O O

OCH3

3. 5.
4. O
O O OH
O OH 1 2 3 4 5

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