A] SPECTROSCOPY

II] Mass Spectrometry (MS)

Basis and Applications
1. Instrumentation and Basis:

2. Modes of Fragmentation:
1) Heterolytic cleavage:

2) Homolytic cleavage:

3. Summary of Fragmentation Processes

1- Saturated hydrocarbons:

1
2- Saturated branched hydrocarbons:

2
- Dimethyldecane:

m/z 57 and m/z 155 are stable 3ary carbocations

- 3,3-Dimethylhexane:

3
3- Unsaturated Hydrocarbons undergo cleavage at α-C-atom: (vinylic cleavage):

α-Carbon (vinylic) or β-C-atom (allylic cleavage in the aliphatic) or (Benzylic cleavage in the aromatic);
allylic and benzylic cleavage are the most common:

1-Butene:

5-Methyl-3-heptene:

4
4- Mc-Lafferty rearrangement:

Involves cleavage of allylic and transfer of the γ-H to the ionized double bond.

Where Q,X,Y and Z may be any combination of C,N,O and S.

1-Pentene:

5
2-Hexene:

Methyl butyrate:

3-Heptene:

6
6- Esters, Aldehydes and Ketones:

α-cleavage):

- Ester:

- Aldehydes:

McLafferty rearrangement for long chain aldehydes:

- Ketones:

Problem: The mass spectrum of a compound known to be either 3-heptanone or its isomer 4-
heptanone is shown to give m/z 43 and m/z 71. Which isomer is most consistent with the data?

7
- Esters: e.g. Methyl butyrate:

OR

N.B.: Any methyl ester may show m/z 31 and m/z 59

Alternatively;

m/z 59 may be formed by the following pathway:

8
Alternatively;

7- Alcohols, Amines and Ethers undergo α, β-cleavage:

a- Alcohols:

- Ethanol:

Mass spectrum of n–pentanol:

Mass spectrum of 2–pentanol:

b- Ethers:

9
di-n-propyl ether:

Ethyl butyl ether:

10
Aromatic ethers:

c- Amines:

Mass spectrum of propylamine: C3H9N (Mass of molecular ion: 59)

Problem: A compound is thought to be either 3-aminooctane OR 4-aminooctane. How to

differentiate?

• Another example:

11
8- Aromatic compounds:

9- Phenols:

12
10- Isotopes in Mass Spectrometry

Fragmentation of Chlorobenzene

MS of chlorobenzene:

Fragmentation of 1-bromopropane:

MS of 1-bromopropane:

13
MS of benzyl chloride:

C6H5 CH2Cl (Mass of molecular ion: 126)

Advantages of MS:

 Rapid identification and quantitation of drugs in body fluids,

 in emergency cases of acute poisoning, is an important application of MS.
 Results are reported within minutes of receipt of few ml of gastric content, blood or urine.
 After separation of samples, spectra are taken at 4s intervals and selected spectra are compared with
reference collection consisting of drug metabolites spectra.
Disadvantages:

 Mass spectrometry is a technique that destroys the sample.

 Spectrometers are expensive.
 Mass spectrometric techniques are very powerful but do not work well for all samples: e.g. positional
isomers.

Molecular weight identification:

 Chemical ionization (CI) is a more effective technique than electron impact (EI) for identifying a
compound’s molecular weight.

 It shows intense ions in the region of a compound’s molecular weight. The pattern of these ions is
characteristic for each reagent gas and is easily recognizable.
14
 Example: An unknown compound from a chemical reaction

 You have treated propenal (acrolein) with HBr in ethane-1,2-diol (or glycol) as solvent for one hour
at room temperature.

 Distillation of the reaction mixture gives a colourless liquid, compound X. What is it?

Fragment at m/z = 73 corresponds to the 5-membered ring structure.

The product is:

15
 III] 1H-NMR SPECTROSCOPY

Basis and Applications

1. Basis of NMR (Nuclear Magnetic Resonance)
Absorption of electromagnetic radiation in the radiofrequency (rf) region by certain nuclei in
organic molecules when they are in a strong magnetic field.

Electromagnetic radiation for use in NMR Spectroscopy: (Radiofrequency [rf] region)

2. Origin of NMR Signal:

Nuclear Spin

Nuclei of many atoms have a special property called NUCLEAR SPIN.

Nuclei having spin rotate about an axis generating magnetic field described by a vector: Nuclear
Magnetic Moment.
The nucleus behaves as a tinny spinning bar magnet because it possesses electrical charge and
mechanical spin. It will generate a small magnetic field.
Nuclei like 1H and 13C have spin quantum number (I) = ½.
They can assume either of 2 spin states: +½ and - ½.
Some other nuclei as 12C, 16O and 32S have no spin; I = 0 and do not give NMR spectrum.

(a) The magnetic field associated with a spinning proton

(b) The spinning proton resembles a tiny bar magnet

16
 15
N
19 14
F N
217
H
F

Atomic mass Atomic Nucleus

I (upper No) number
(lower No)

1 17 15
H, O, N,
Half integer Odd Odd or 1 8 7
(Fraction) Even 13
C
6

Integer Even Odd 2 14

H, N
(Whole) 1 7

Zero Even Even 12 16 34

C, O, S
6 8 16

NMR can differentiate between:

Different ISOTOPES (1H, 2H) and

Different ELEMENTS (1H, 13C)
As they absorb energy at different magnetic field strength and radiofrequency.

We can obtain NMR spectrum for each element without interference from others

Effect of an External Magnetic Field

In absence of a magnetic field, magnetic moments of protons in a given sample are randomly
oriented.
When the sample is placed in an applied external magnetic field, protons, like magnet bars, will
respond to the influence of that applied magnetic field and will tend to align themselves.
They may assume one of 2 possible orientations with respect to external applied magnetic field.
The magnetic moment of the proton nuclei may be:
Aligned with the external magnetic field
Aligned against external field.
17
 When aligned WITH field:

 Spin state is + ½, α-spin.

 It is of lower energy state
 It is described as parallel with the field

When aligned AGAINST the field:

 Spin state is - ½, β-spin.

 It is of higher energy state
 It is described as antiparallel with the applied field

Bo

(a) In the absence of an external magnetic field, the nuclear spins of the protons are randomly oriented.
(b) In the presence of an external magnetic field Bo, the nuclear spins are oriented so that the resulting
nuclear magnetic moments are aligned either parallel antiparallel to Bo. The lower energy orientation
is the one parallel to Ho and there are more nuclei that have this orientation.

18
Spin Flipping

When exposed to the proper frequency of radiowaves, magnetic moments of a fraction of parallel
proton nuclei absorb energy and turn around or FLIP, to the higher energy antiparallel state. Converted to the
β-spin, nuclei lose absorbed energy to its surrounding as heat and return to low energy, α-spin state (parallel).

Spin Flipping

When exposed to the proper frequency of radiowaves,

magnetic moments of a fraction of parallel proton nuclei absorb
energy and turn around or FLIP, to the higher energy
antiparallel state. Converted to the β-spin, nuclei lose absorbed
energy to its surrounding as heat and return to low energy, α-
spin state (parallel).

α β

hv
Bo Bo
Radiofrequency region

Change of nuclear spin state

Nucleus aligned against applied magnetic field

Nucleus aligned with applied magnetic field low energy level

3. Resonance:
Placing a compound containing 1H in a very strong magnetic field and irradiating it with
electromagnetic energy, nuclei absorb E; and when the particular combination of external magnetic field (Bo)
and radiofrequency is achieved, this will cause proton to flip from parallel to antiparallel state. Protons are
said to be in RESONANCE. Thus, Nuclear Magnetic Resonance means: Nuclei in resonance in a magnetic
field.

19
 4. Relationship of External Magnetic Field Strength with Energy Difference between
α- and β-Spin

An external magnetic field causes the 2 nuclear spin states to have different energies. The difference
in energy ΔE (amount of E required to flip magnetic moment from parallel to antiparallel states) is
proportional to the strength of the applied field Bo.
The greater the strength of the applied magnetic field, the greater is the difference in energy between
the α- and the β-spin states. The nucleus become more resistant to being flipping and higher E, high
frequency radiation is required.

5. Instrumentation:
Types of NMR spectrometers:

1. Sweep (continuous wave, CW) spectrometers.

2. Fourier-Transform (FT) NMR spectrometers

Diagram of an NMR Spectrometer

FT-NMR Spectrometers

They use magnets having higher magnetic field strength than CW spectrometers.
They employ computers allowing signal analysis by mathematical calculation known as Fourier
Transform.
They accumulate data scans and average them.
20
Advantages of FT-NMR Spectrometers

1. Can generate NMR spectrum in few seconds (5s).

2. Store large number of repetitive scans in computer memory.
3. Noise is cancelled; thus intensities of real signals are increased.
4. Very high resolution and greater sensitivity.
5. Sample is irradiated with short pulse rf radiation which excites all nuclei at once.
6. Data are collected in function of time and transformed by computer into function of frequency before
individual peaks can be identified.

6. 1H NMR Spectrum:
Peak Position: Chemical Shift

Signals for different protons occur at different positions in an NMR spectrum: They have different
CHEMICAL SHIFTS.
The chemical shift depends on the magnetic environment of each proton.
This magnetic environment depends on:
a) Magnetic field generated by circulation of electrons (nature of bonds).

b) Magnetic field resulting from other nearby protons (number and types of protons).

Chemical shifts are reported in δ values and expressed as ppm of applied radiofrequency:
- At 60 MHz, 1 ppm is 60 Hz.

- At 200 MHz, 1 ppm is 200 Hz.

Chemical shift is expressed downfield from the position of absorption of a reference TMS which is
set at 0 ppm (δ 0). It allows calibration of the instrument.

Chemical Shift Calculation

Chemical shift of a proton is the distance of the signal from the reference in Hz. It is proportional to the
strength of external magnetic field.

e.g. The chemical shift for CHCl3 proton in ppm = 7.86 ppm

21
 The 200-MHz 1H NMR spectrum of chloroform (HCCl3).

Chemical shifts are measured along the x-axis in parts per million (ppm) from TMS as the reference,
which is assigned a value of zero.

Tetramethylsilane (TMS):
Advantages as internal standard:

1. A small amount is added to the sample whose spectrum is measured.

2. It gives an intense sharp signal even at low concentration; having 12 protons in magnetically
equivalent environment.
3. Silicon is electron-positive relative to C and tends to donate its electrons to CH3 groups; thereby
increasing their shielding.
4. Highly shielded protons of TMS resonate upfield from most other protons. This signal does not
interfere with other protons signals.
5. TMS is chemically inert.
6. Has low b.p. ≈ 26.5oC; so can be easily removed from sample by evaporation.
7. Soluble in most used solvents.

1
HNMR spectrum of CH3OH (Solvent CCl4)

22
 1HNMR spectrum of p-Xylene

Shielding and Deshielding:

Shielding is caused by circulating σ electrons.

It depends on the relative electron density around the proton.
Electron density depends on presence or absence of electronegative groups withdrawing electrons
from C-H bond.
Proton that flips more easily absorbs E at lower Bo and gives rise to absorption peak downfield (to
the left, deshielded).
Proton that flips with difficulty absorbs E at higher Bo and gives rise to absorption peak upfield (to
the right , shielded).
A shielded proton absorbs at higher external magnetic field strength (to the right).
The external magnetic field should be made larger by the spectrometer in order to compensate for the
large induced field.
A deshielded proton absorbs at low external magnetic field strength (to the left).
The external magnetic field should be made smaller by the spectrometer in order to compensate for
the small induced field.

23
Integration (Relative Peak Area):
NMR spectrum can predict the number of magnetically different protons in a molecule.
Amount of E absorbed at each resonance frequency is proportional to the number of nuclei that are
absorbing at that frequency.
The areas under each signal are in the ratio of the number of Hs in each part of the molecule.
Ratio of the heights of different integral lines is equal to the ratio of number of protons giving rise to
that peaks.

Ringing:

- It is a fluctuation to the right of each signal and arises from distortion of magnetic field when the
sample is scanned rapidly.
- It is an indication of the proper adjustment of the instrument.

Induced Magnetic Field:

The chemical shift of a signal is related to electron densities and electron circulations in the molecule.
Under the influence of an external magnetic field, electrons move in certain paths.
Because electrons are charged particles, they generate tiny magnetic fields.
The small magnetic field generated by electrons is called: INDUCED FIELD.

24
 Circulation of electrons of a C-H bond under the influence of an external magnetic field. Electron
circulations generate a small magnetic field (an induced field) that shields the proton from the external
field. The magnetic field felt by proton is slightly less than Bo. Bo should be increased.

7. Factors Affecting Chemical Shift:

1. Electronegativity (Inductive Effect)

25
2. Anisotropic Effects Field Induced by π Electrons

Molecular magnetic fields generated by π electrons are DIRECTIONAL (UNSYMMETRICAL).

Measurement that varies depending on the direction in which measurement is taken is said to be:
ANISOTROPIC.
Anisotropic effect is contrasted to IE which is SYMMETRICAL (non directional) around the proton.

a) Benzene ring:
π electrons are delocalized around the ring.
Under the influence of an external magnetic field, π electrons circulate around the ring.
This circulation, called ring current, generates a molecular magnetic field.
Near the protons, the field generated by the ring current is in the same direction as the applied
external magnetic field Bo.
Less external field Bo is needed to bring these protons into resonance compared with that needed for
alkyl protons.
Aryl-Hs are deshielded and absorb far downfield at δ 7.22 ppm.

26
 Br
H3C CH3

H H
CH3

b) Vinylic and Aldehydic Protons:

Vinylic and Aldehhydic protons are also deshielded by anisotropic effect.

π electrons are set in motion within the bond and generate molecular field that adds to the applied
field near =C-H.
Protons come to resonance at higher δ values.

c) Alkyne Protons:
Alkyne protons appear ~ δ 1.5 – 3.5 ppm.
Electron circulate on around triple bond occurs such that proton experiences a diamagnetic shielding
effect.
When axis of alkyne group lies PARALLEL to direction of Bo, π electrons are induced to circulate
around axis and the resultant magnetic field acts in a direction opposing Bo near H.
H experiences lower values of field which should be increased (appears at low δ values) (upfield).

27
Offset Scan:
Absorption that is far downfield outside the normal scan for some spectrometers.

1H NMR spectrum of Z-glycine

1H NMR spectrum of Methyl acetoacetate (keto ester) In CDCl3

28
8. Relative Positions of Proton Absorption:

29
30
9. Chemical Shift Equivalent and Non-equinalent Protons:

Magnetically equivalent = chemically equivalent = chemical shift equivalent

Give only ONE 1H NMR signal
Equivalent protons NEED NOT necessarily be on the same carbon.
Et2O contains 2 types of homotopic protons: CH3 and CH2

H3C-H2C O CH2CH3

10. Signal Splitting, Spin-spin Coupling

H3C Cl
C
H2

31
 Peaks are resolved or SPLIT into groups of peaks.
This splitting (spin-spin splitting) is caused by the presence of neighboring protons (on adjacent C)
that are NON EQUIVALENT.
Protons that split each other’s signals are said to have undergone: Spin-spin coupling.
Splitting is NOT observed for chemically equivalent (homotopic protons) or enantiotopic protons, e.g.
CH3-CH3.
Spin-spin coupling effects are transferred through bonding electrons and are not observed if coupled
protons are separated by more than 3 σ bonds.
CH3

H3C C O CH3

CH3

Reason for Spin-spin coupling of Protons

Splitting of signal arises from the 2 spin states (parallel and antiparallel) of the neighboring protons.
Consider the case of ONE non equivalent proton: Spinning of protons generate a magnetic moment.

Hb is parallel Hb is antiparallel
Its moment adds to Bo Its moment is substracted from

Ha Hb Ha Hb
Bo
C C C C

Ha and Hb are NON equivalent

If the spin of Hb proton is parallel, its magnetic moment adds to the applied magnetic field; Ha sees a
slightly stronger field and comes into resonance at slightly lower applied field strength (to the left).
If Hb is in antiparallel state, its magnetic moment decreases the magnetic field around Ha. It takes
slightly more applied Bo for Ha to come into resonance (to the right).
Because about ½ Hb nuclei are parallel and ½ are antiparallel, there are 2 types of Ha in the sample:
Ha with neighboring Hb in parallel spin state
Ha with neighboring Hb in antiparallel spin state
2 peaks are observed for Ha.

Possible magnetic
Orientations
α, β J

32
 11. Multiplicity Rule: n + 1 rule:

We can predict the number of spin-spin splitting peaks by counting the number (n) of
neighboring protons NON EQUIVALENT to the proton in question and adding 1.
a b
H3C CH2 Cl

• For (a) protons:

3 equivalent protons (a) see 2 neighboring NON equivalent (b) protons.
Their signal is split into 2 + 1 = 3 lines.
• For (b) protons:
2 equivalent protons (b) see 3 neighboring NON equivalent (a) protons.
Their signal is split into 3 + 1 = 4 lines.

Toluene:
Toluene has 4 groups of chemically non equivalent protons; However, it shows ONLY 2
absorption peaks.
* 1 for CH3 * 1 for ring protons
Reason:
Magnetic environment of non equivalent ring protons are so similar that protons exhibit identical
chemical shift. No splitting is observed.
H H

H CH3

H H

12. Relative Intensities (Pascal Triangle):

33
For HA = 2 neighboring Hs, 2 + 1 = 3 lines; For HX = 1 neighboring H, 1 + 1 = 2 lines

13. Splitting Pattern And Spin-Spin Splitting Diagrams:

Singlet (s):
Proton with NO neighboring, non equivalent protons shows a single peak called a singlet (s).

Doublet (d):
Proton with ONE neighboring NON equivalent proton gives signal split into a double line called
doublet (d).

J constant is the coupling constant. Jab means coupling cte for Ha split by Hb or coupling cte for Hb
split by Ha. For a pair of coupled protons, J value is the same in each of the 2 doublets
J values measured on instrument operating at 60 MHz will be the SAME as those measured on
instrument operating at 300 MHz.
Reason: Magnitudes of coupling constant are NOT dependent on magnitude of applied field.
Coupling is caused entirely by Internal Forces.
34
Tree Diagram for (d):
Splitting trees are read from top to bottom; Lines represent absorption peaks: height is
proportional to area under the peak.

Ha
Ha Hb

C C

Jab

Triplet (t):
Proton with Two neighboring NON equivalent proton gives signal split into a triple line called triplet
(t).
2 equivalent protons on adjacent C split the signal of an absorbing proton into 2 + 1 = 3 peaks or
triplet (t) in a proportion of 1:2:1.

35
Spins of neighboring Hb will affect the magnetic field felt by Ha

Possible combinations of spins for 2 Hb protons are:

Both Hb protons may be parallel to the applied field Bo and increase field felt by Ha.
Both Hb protons may be antiparallel to the applied field Bo and decrease field felt by Ha.
One Hb proton is parallel and one Hb proton is antiparallel having no effect upon field felt by Ha.
Because parallel and antiparallel combinations are twice as probable to occur, we obtain ratio 1:2:1
for the 3 peaks in the triplet.

Quartet (q):

Proton with Three neighboring NON equivalent proton gives signal split
into a quadruple line called quartet (q).

Ha Hb

C C Hb

Hb

36
37
300 MHz 1H NMR spectrum of ethyl bromide.

38
39
 200 MHz 1H NMR spectrum of 2,3,4-trichloroanisole

Br-CH CH CH CH
2 2 2 3

40
Br-CH CH CH CH
2 2 2 3

300 MHz 1H NMR spectrum of 1-bromobutane

90 MHz 1H NMR spectrum of 2-heptanone

500 MHz 1H NMR spectrum of 2-heptanone

1
H NMR spectrum of phenethyl acetate

41
14. Chemical Exchange:

Protons attached to electronegative atoms with lone pairs as O, N, S undergo rapid Chemical
Exchange.
is found in 1H NMR spectra of alcohols, amines, carboxylic acids.

42
Protons undergoing rapid chemical exchange can be easily detected by placing the compound in D2O
(deuterium oxide).
Protons are rapidly replaced by deuterons and proton signal disappears from the spectrum.

H-bond:
Chemical shifts of OH, NH protons are solvent, temperature and concentration dependent because of
H-bonding.
H-bonding decreases electron density around proton moving proton peak to higher frequency
(deshielding).
Decrease in concentration in polar solvent disrupts H-bonding and the peak appears at lower
frequency (shielding). Alcohol molecules become less polymeric.
Increase in temperature has a similar effect.

R O H O H O H

R R

43
 IV] 13C NMR SPECTROSCOPY

Basis and Applications

12
C nucleus is not magnetically active (spin number I = 0), but 13C nucleus has I = ½. Natural abundance
of 13C is only 1.1% that of 12C and its sensitivity is about 1.6% that of 1H, overall sensitivity of 13C with 1H is
about 1/5700.

Theory:
Aspects of 13C NMR differing from 1H NMR are:

- In CPD (broadband proton decoupled) 13C spectrum, peaks are singlets unless molecule contains other
magnetically active nuclei as 31P or 19F.
13
- C peaks are distributed over a larger chemical-shift range in comparison with the proton range.
13
- C peak intensities do not correlate with the number of carbon atoms in a given peak in spectra, due to
longer T1 values.
13
- C nuclei are less abundant and less sensitive than protons. Larger samples and longer times are
needed.
- For a given solvent, 13C and 1H solvent peaks differ in multiplicities.

1
H decoupling techniques:

 13
C nucleus does not show coupling in 1H NMR spectra due to low natural abundance of 13C (1.1%).
 1
H nucleus is > 99% in natural abundance and couples to 13C nuclei.
 Because of large 1JCH values for 13C-1H (~110- 320 Hz) and appreciable 2JCH, 3JCH values for 13C-C-1H
and 13C-C-C-1H (~ 0-60 Hz) couplings, proton coupled 13C spectra show complex overlapping
multiplets difficult to interpret.

1
H coupled 13C spectrum of cholesterol in CDCl3 at 150.9 MHz

44
  To solve this problem, the use of proton broadband decoupling-irradiation and detection of 13C signals
was developed. Irradiation of protons over a broad frequency range by means of a broadband generator
or with Composite Pulse Decoupling (CPD) removes these couplings.

1
H decoupled 13C spectrum of cholesterol in CDCl3 at 150.9 MHz

 Spectrum of cholesterol shows 27 single peaks, each representing one of the carbon atoms.
 When 13C is decoupled from 1H, useful information on multiplicity of the carbon is lost (n + 1 rule).
Example: Me group is quartet (3 +1) and a methine group is doublet (1 +1).

Chemical shift scale and range:

 Scale is in δ units (ppm).

 Chemical shifts in 13C spectra range over ~ 220 ppm from TMS; ~ 20 times that 1H spectra (~ 10 ppm).
 As result of large range and sharpness of decoupled peaks, coincidence of 13C chemical shifts are uncommon
and impurities are readily detected.

45
 In general, in comparison with 1H NMR spectra, it is more difficult to correlate 13C shifts with substituent
electronegativity.
 13C peaks do not correlate with the ratio of C atoms in spectra.

46
Sensitivity
13
C nuclei are less abundant and less sensitive than protons. Larger samples and longer times are needed
to acquire sufficient signal strength.

Solvents:

Samples for 13C NMR spectrometry are dissolved in CDCl3 and 13C peak of TMS is used as internal
reference.

Interpretation of diethyl phthalate 13C spectrum:

It is possible by inspection of 13C spectrum to recognize nuclei that do not bear protons by their low
intensity: C=O and peak labeled (1). Non-protonated C atoms have longer T1 relaxation times, which results
in small peaks. It is therefore possible to recognize C=O groups, CN, non protonated alkene and alkyne C
atoms and other quaternary C atoms readily.
Diethyl phthalate C12H14O4 has an axis of symmetry and a plane of symmetry: decoupled 13C spectrum consists
of 6 peaks. Since there is no coupling, peaks are singlets and there is no overlap.

Diethyl phthalate 13C decoupled spectrum at 150.9 MHz in CDCl3

Diethyl phthalate 13C coupled spectrum at 150.9 MHz in CDCl3

47
 Alkyl groups are placed on the right-hand side of the spectrum with deshielded CH2 group to the left
of CH3 group. It is easy to designate the strongly deshielded cluster of 3 peaks as aromatic.
C=O group is on the far left
13
C aromatic nucleus (1) without attached proton gives a peak distinctly decreased in height. The same applies
to C=O group.
1
J coupling in (b) provides multiplicities i.e. number of protons directly attached. These couplings confirm
chemical shift assignments: From left to right: q (CH3 group), t (CH2 group). Then, there is a cluster of 2 d of
2 aromatic CH groups and s of C atom with no proton (1). Finally, C=O singlet at high frequency.

Problems:

1
H decoupled 13C spectrum of t-butanol at 75.5 MHz in CDCl3

(a) 1H decoupled 13C spectrum of 2,2,4-trimethyl-1,3-pentanediol in CDCl3 at 75.5 MHz

(b) same at 150.9 MHz

48
 Individual 13C signals are weak because isotope of carbon (13C) that gives rise to 13C NMR signals
constitutes only 1.11% of carbon atoms. The most abundant isotope of carbon, 12C has no nuclear spin and
therefore cannot produce an NMR signal. The low abundance of 13C means that the intensities of signals in 13C
NMR compared with those in 1H NMR are reduced by a factor of approximately 100. Therefore, the overall
intensity of a signal is about 6400 times less than the intensity of 1H signal.

One advantage to 13C NMR spectroscopy is that chemical shifts range over about 220 ppm, compared with
about 12 ppm for 1H NMR. This means that signals are less likely to overlap.

A disadvantage of 13C NMR spectroscopy is that, unless special techniques are used, the area under 13C
NMR signal is not proportional to the number of atoms giving rise to the signal. Thus, the number of carbons
giving rise to 13C NMR signal cannot be determined by integration.

2-Butanol has carbons in four different environments, so there are 4 signals in the spectrum. Relative
positions of signals depend on the same factors that determine relative positions of proton signals in 1H NMR.
Carbons in electron-dense environments produce low-frequency signals, and carbons close to electron-
withdrawing groups produce high-frequency signals. This means that signals for carbons of 2-butanol are in
the same relative order that expected for signals of protons on those carbons in 1H NMR spectrum. Thus, carbon
of methyl group farther away from electron-withdrawing OH group gives the lowest-frequency signal. As
frequency increases, other methyl carbon comes next, followed by methylene carbon; and carbon attached to
OH group gives the highest-frequency signal.

Proton decoupled 13C NMR spectrum of 2-butanol

Signals are not normally split by neighboring carbons because there is little likelihood of an adjacent
carbon being 13C. The probability of 2 13C carbons being next to each other is 1.11% (about 1 in 10,000).
(Because 12C does not have a magnetic moment, it cannot split the signal of an adjacent 13C.

49
 Proton coupled 13C NMR spectrum of 2-butanol: splitting is observed

Problems
Identify the compound with molecular formula C9H10O2 that gives the following 13C NMR spectrum:

Identify each compound from its molecular formula and its 13C NMR spectrum.

50
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