SPECTROSCOPIC METHODS

OF ORGANIC MOLECULES

Professor Ioannis P. Gerothanassis

Department of Chemistry, University of Ioannina
FEBRUARY 2018
 SPECTROSCOPIC METHODS
OF ORGANIC MOLECULES

UV SPECTROSCOPY

Professor Ioannis P. Gerothanassis

Department of Chemistry, University of Ioannina
 WHAT IS SPECTROSCOPY?
The study of the interaction between
ELECTROMAGNETIC (EM) RADIATION and MATTER

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 SPECTROSCOPIC ANALYSIS

covers

ATOMIC MOLECULAR
SPECTROSCOPY SPECTROSCOPY

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 To Understand Spectroscopy We
Must Understand Electromagnetic
Radiation
What is Electromagnetic Radiation?
• is a form of energy that has both Wave and Particle
Properties.
• For example: Ultraviolet, visible, infrared, microwave, radio
wave.

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 WAVE PROPERTIES
• EM radiation is conveniently modeled as waves consisting of
perpendicularly oscillating electric and magnetic fields, as
shown below.
y

x
z Electric Field

Magnetic Field

Direction of
propagation
6

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 o At 90° to the direction of propagation is an
oscillation in the ELECTRIC FIELD.

o At 90° to the direction of propagation and 90°

from the electric field oscillation (orthagonal)
is the MAGNETIC FIELD oscillation.

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 Wave parameters
+ Wavelength ()
Electric Field

Amplitude (A)

- Time or Distance

We Use Symbols to Designate the Various Properties of

Waves
  is the wavelength of the waves
 is the frequency of the waves
 c is the speed of light 8

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 Definitions:
 Period (p) – the time required for one cycle to pass a fixed point in
space.

 Frequency () – the number of cycles which pass a fixed point in

space per second.

 Amplitude (A) – The maximum length of the electric vector in the

wave (Maximum height of a wave).

 Wavelength () – The distance between two identical adjacent points

in a wave (usually maxima or minima).

9
 Wavenumber () - The number of waves per cm in units of cm-1.

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• Purpose of each Electromagnetic Radiation
UV Spectroscopy

The Spectroscopic Process

 In UV spectroscopy, the sample is irradiated with the broad spectrum
of the UV radiation
 If a particular electronic transition matches the energy of a certain
band of UV, it will be absorbed
 The remaining UV light passes through the sample and is observed
 From this residual radiation a spectrum is obtained with “gaps” at
these discrete energies – this is called an absorption spectrum






11


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UV Spectroscopy
Observed electronic transitions
 Here is a graphical representation

s
Unoccupied levels


Atomic orbital Atomic orbital

Energy n

Occupied levels


s
Molecular orbitals 12

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UV Spectroscopy
Observed electronic transitions
 From the molecular orbital diagram, there are several possible
electronic transitions that can occur, each of a different relative
energy:

s
s s alkanes

s  carbonyls

  unsaturated cmpds.
Energy
n
n s O, N, S, halogens

n  carbonyls


s
13

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UV Spectroscopy
Observed electronic transitions
 Although the UV spectrum extends below 100 nm (high energy),
oxygen in the atmosphere is not transparent below 200 nm

 Special equipment to study vacuum or far UV is required

 Routine organic UV spectra are typically collected from 200-700

nm

 This limits the transitions that can be observed:

s s alkanes 150 nm

s  carbonyls 170 nm

  unsaturated cmpds. 180 nm √ - if conjugated!

n s O, N, S, halogens 190 nm

n  carbonyls 300 nm √ 14

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 Electromagnetic Radiation

Violet0 400 - 420 nm Yellow 570 - 585 nm

Indigo 420 - 440 nm Orange 585 - 620 nm
Blue 440 - 490 nm Red 620 - 780 nm
Green 490 - 570 nm

S. Damkondwar, Pune, Maharashtra, India, January, 2013

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 http://www.math.andyou.com/310

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UV Spectroscopy

From an experimental point of view, three other

considerations must be made:
 a longer path length, l, through the
sample will cause more UV light to be
absorbed – linear effect

 the greater the concentration, c, of the

sample, the more UV light will be
absorbed – linear effect

 some electronic transitions are more

effective at the absorption of photon than
others – molar absorptivity, e,
this may vary by orders of magnitude… 19

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UV Spectroscopy
The Beer-Lambert Law: A=ecl

 for most UV spectrometers, l would remain constant (standard

cells are typically 1 cm in path length)

 Concentration c is typically varied depending on the strength of

absorption observed or expected – typically dilute – sub .001 M

 molar absorptivities ε vary by orders of magnitude:

• values of 104-106 are termed high intensity absorptions
• values of 103-104 are termed low intensity absorptions
• values of 0 to 103 are the absorptions of forbidden
transitions

A is unitless, so the units for e are cm-1 x M-1 and are rarely
expressed
Since path length and concentration effects can be easily factored out,
absorbance simply becomes proportional to e, and the y-axis is
expressed as e directly or as the logarithm of e 20

V. Krishnakumar, Prof. Depart. of Physics, Periyar University, Salem. Tamilnadu, India

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UV Spectroscopy

Selection Rules
 Not all transitions that are possible are observed

 For an electron to transition, certain quantum mechanical

constraints apply – these are called “selection rules”

 For example, an electron cannot change its spin quantum

number during a transition – these are “forbidden”
Other examples include:
• the number of electrons that can be excited at
one time
• symmetry properties of the molecule
• symmetry of the electronic states

 To further complicate matters, “forbidden” transitions are

sometimes observed (albeit at low intensity) due to other
factors 21

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UV Spectroscopy
Instrumentation and Spectra

 Here is a simple schematic that covers most modern UV

spectrometers:

log(I0/I) = A
I0 I

sample
UV-VIS sources

200 700

detector
, nm

monochromator/
reference

beam splitter optics I0 I0

22

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UV Spectroscopy

Instrumentation and Spectra,

 Two sources are required to scan the entire UV-VIS band:

• Deuterium lamp – covers the UV – 200-330
• Tungsten lamp – covers 330-700

 As with the dispersive IR, the lamps illuminate the entire band
of UV or visible light; the monochromator (grating or prism)
gradually changes the small bands of radiation sent to the
beam splitter

 The beam splitter sends a separate band to a cell containing

the sample solution and a reference solution

 The detector measures the difference between the transmitted

light through the sample (I) vs. the incident light (I0) and
sends this information to the recorder
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UV Spectroscopy
Instrumentation – Sample Handling

 Solvents must be transparent in the region to be observed; the

wavelength where a solvent is no longer transparent is referred to as
the cutoff

 Since spectra are only obtained up to 200 nm, solvents typically only
need to lack conjugated  systems or carbonyls

Common solvents and cutoffs:

acetonitrile 190
chloroform 240
cyclohexane 195
1,4-dioxane 215
95% ethanol 205
n-hexane 201
methanol 205
isooctane 195
water 190 24

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UV Spectroscopy
Instrumentation – Sample Handling

 Additionally solvents must preserve the fine structure (where it is

actually observed in UV!) where possible

 H-bonding further complicates the effect of vibrational and rotational

energy levels on electronic transitions, dipole-dipole interacts less so

 The more non-polar the solvent, the better (this is not always
possible)

25

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UV Spectroscopy
Band Structure

 When these energy levels are superimposed, the effect can be

readily explained – any transition has the possibility of being
observed

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 MOLECULAR ABSORPTION SPECTRA

 The sharpness of molecular

absorption spectra also depends on
the state of the sample.
 Figure (a) shows an absorption
band due to transitions between
electronic-vibrational-rotational
states
 Figure (c) shows a continuous
spectra due to the sample is in the
condensed state. In condensed
states the spectra broaden due to
molecular collisions.

27

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UV Spectroscopy

 Due to the lack of any fine structure, spectra are rarely

shown in their raw form, rather, the peak maxima are simply
reported as a numerical list of “lamba max” values or max

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UV Spectroscopy
Chromophores
1. Alkanes – only posses s-bonds and no lone pairs of electrons, so
only the high energy s  s* transition is observed in the far UV

This transition is destructive to the molecule, causing cleavage of

the s-bond

C C
s

s C C

29

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UV Spectroscopy
Chromophores
2. Alcohols, ethers, amines and sulfur compounds – in the cases of
simple, aliphatic examples of these compounds the n  s* is the
most often observed transition; like the alkane s  s* it is most
often at shorter  than 200 nm

Note how this transition occurs from the HOMO to the LUMO

C N
sCN

C N

nN sp 3
C N anitbonding
orbital

30
sCN C N

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UV Spectroscopy
Chromophores
3. Alkenes and Alkynes – in the case of isolated examples of these
compounds the   * is observed at 175 and 170 nm, respectively

Even though this transition is of lower energy than s  s*, it is still

in the far UV – however, the transition energy is sensitive to
substitution




31

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UV Spectroscopy

Chromophores
4. Carbonyls – unsaturated systems incorporating N or O
can undergo n  * transitions (~285 nm) in addition
to   *

Despite the fact this transition is forbidden by the

selection rules (e = 15), it is the most often observed
and studied transition for carbonyls

This transition is also sensitive to substituents on the

carbonyl

Similar to alkenes and alkynes, non-substituted

carbonyls undergo the   * transition in the vacuum
UV (188 nm, e = 900); sensitive to substitution effects
32

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UV Spectroscopy
Chromophores
4. Carbonyls – n  * transitions (~285 nm);   * (188 nm)


It has been determined
from spectral studies,
n C O
that carbonyl oxygen
more approximates sp
rather than sp2 !
 O

sCO transitions omitted for clarity

33

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Forbidden n  π* transitions

34
UV Spectroscopy
Chromophores - Substituent Effects
General – Substituents may have any of four effects on a chromophore
i. Bathochromic shift (red shift) – a shift to longer ; lower
energy

ii. Hypsochromic shift (blue shift) – shift to shorter ; higher

energy

iii. Hyperchromic effect – an increase in intensity

iv. Hypochromic effect – a decrease in intensity

Hyperchromic

e
Hypsochromic Bathochromic
Hypochromic

35

200 nm 700 nm
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UV Spectroscopy
Chromophores - Substituent Effects
1. Conjugation – most efficient means of bringing about a
bathochromic and hyperchromic shift of an unsaturated
chromophore:
H2C
max nm e
175 15,000
CH2

217 21,000

258 35,000

465 125,000
-carotene

O
n  * 280 12
  * 189 900

O n  * 280 27 36
  * 213 7,100

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UV Spectroscopy
Chromophores - Substituent Effects
1. Conjugation – Alkenes
The observed shifts from conjugation imply that an increase in
conjugation decreases the energy required for electronic excitation
From molecular orbital (MO) theory two atomic p orbitals, f1 and f2
from two sp2 hybrid carbons combine to form two MOs Y1 and Y2*
in ethylene

Y 2

f1 f2

Y1  37

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UV Spectroscopy
Chromophores - Substituent Effects
2. Conjugation – Alkenes
When we consider butadiene, we are now mixing 4 p orbitals giving
4 MOs of an energetically symmetrical distribution compared to
ethylene

Y 4
Y 2

Y 3

Y2

Y1 
Y1

38
DE for the HOMO  LUMO transition is reduced

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UV Spectroscopy
Chromophores - Substituent Effects
2. Conjugation – Alkenes
Extending this effect out to longer conjugated systems the energy
gap becomes progressively smaller:

Energy Lower energy =

Longer wavelengths

ethylene
butadiene 39
hexatriene
octatetraene
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UV Spectroscopy
Chromophores - Substituent Effects
2. Conjugation – Alkenes
Similarly, the lone pairs of electrons on N, O, S, X can extend
conjugated systems – auxochromes
Here we create 3 MOs – this interaction is not as strong as that of a
conjugated -system

Y 3

 Y2
Energy


nA

40
Y1

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UV Spectroscopy
Structure Determination
Dienes
1. General Features
For acyclic butadiene, two conformers are possible – s-cis and s-
trans

s-trans s-cis

The s-cis conformer is at an overall higher potential energy than

the s-trans; therefore the HOMO electrons of the conjugated
system have less of a jump to the LUMO – lower energy, longer
wavelength

41

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UV Spectroscopy
Structure Determination
Dienes
1. General Features
Two possible   * transitions can occur for butadiene Y2  Y3
and Y2  Y4*
Y4
175 nm –forb. 175 nm
Y3
217 nm 253 nm
Y2
s-trans s-cis
Y1

The Y2  Y4* transition is not typically observed:

• The energy of this transition places it outside the region
typically observed – 175 nm

• For the more favorable s-trans conformation, this

transition is forbidden 42
The Y2  Y3* transition is observed as an intense absorption

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UV Spectroscopy
Structure Determination
Dienes
2. Woodward-Fieser Rules - Dienes
The rules begin with a base value for max of the chromophore
being observed:

acyclic butadiene = 217 nm

The incremental contribution of substituents is added to this base

value from the group tables:
Group Increment
Extended conjugation +30
Each exo-cyclic C=C +5
Alkyl +5
-OCOCH3 +0
-OR +6
-SR +30
43
-Cl, -Br +5
-NR2 +60

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UV Spectroscopy
Structure Determination
Aromatic Compounds
1. General Features
Although aromatic rings are among the most widely studied and
observed chromophores, the absorptions that arise from the
various electronic transitions are complex

On first inspection, benzene has six -MOs, 3 filled , 3 unfilled *

6

4 5

2 3
44
1
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UV Spectroscopy
Structure Determination
Aromatic Compounds
1. General Features
One would expect there to be four possible HOMO-LUMO   *
transitions at observable wavelengths (conjugation)

Due to symmetry concerns and selection rules, the actual transition

energy states of benzene are illustrated at the right:
E1u
6
B1u
200 nm
4 5 (forbidden)
B2u

180 nm
260 nm (allowed)
2 3 (forbidden)
A1g 45
1

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UV Spectroscopy
Structure Determination
Aromatic Compounds
1. General Features
The allowed transition (e = 47,000) is not in the routine range of UV
obs. at 180 nm, and is referred to as the primary band

The forbidden transition (e = 7400) is observed if substituent effects

shift it into the obs. region; this is referred to as the second primary
band

At 260 nm is another forbidden

transition (e = 230), referred to
as the secondary band.

This transition is fleetingly allowed

due to the disruption of symmetry
by the vibrational energy states,
the overlap of which is observed
in what is called fine structure 46

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UV Spectroscopy
Structure Determination
Aromatic Compounds
2. Substituent Effects
a. Substituents with Unshared Electrons
• If the group attached to the ring bears n electrons,
they can induce a shift in the primary and secondary
absorption bands

• Non-bonding electrons extend the -system through

resonance – lowering the energy of transition   *

• More available n-pairs of electrons give greater shifts

G G G G

47

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UV Spectroscopy
Structure Determination
Aromatic Compounds
2. Substituent Effects
a. Substituents with Unshared Electrons
• pH can change the nature of the substituent group
• deprotonation of oxygen gives more available n-pairs,
lowering transition energy
• protonation of nitrogen eliminates the n-pair,
raising transition energy
Primary Secondary
Substituent max e max e
-H 203.5 7,400 254 204
-OH 211 6,200 270 1,450
-O- 235 9,400 287 2,600
-NH2 230 8,600 280 1,430
-NH3+ 203 7,500 254 169
-C(O)OH 230 11,600 273 970
48
-C(O)O- 224 8,700 268 560

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UV Spectroscopy
Structure Determination
Aromatic Compounds
2. Substituent Effects
d. Di-substituted and multiple group effects
• With di-substituted aromatics, it is necessary to
consider both groups

• If both groups are electron donating or withdrawing,

the effect is similar to the effect of the stronger of the
two groups as if it were a mono - substituted ring

• If one group is electron withdrawing and one group

electron donating and they are para - to one another,
the magnitude of the shift is greater than the sum of
both the group effects

• Consider p - nitroaniline:
O O
H2N N H2N N
49
O O

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UV Spectroscopy
Structure Determination
Aromatic Compounds
2. Substituent Effects
d. Polynuclear aromatics
• When the number of fused aromatic rings increases,
the  for the primary and secondary bands also
increase

• For heteroaromatic systems spectra become complex 50

with the addition of the n  * transition and ring size
effects and are unique to each case
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UV Spectroscopy
Visible Spectroscopy
Color
1. General
• When white (continuum of ) light passes through, or is
reflected by a surface, those ls that are absorbed are
removed from the transmitted or reflected light respectively

• What is “seen” is the complimentary colors (those that are

not absorbed)

• This is the origin of the “color wheel”

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UV Spectroscopy
Visible Spectroscopy
Color
1. General
• Organic compounds that are “colored” are typically those with
extensively conjugated systems (typically more than five)

• Consider -carotene

-carotene, max = 455 nm

max is at 455 – in the far blue region

of the spectrum – this is absorbed

The remaining light has the

complementary color of orange
52

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UV Spectroscopy
Visible Spectroscopy
Color
1. General
• Likewise:

lycopene, max = 474 nm

O
H
N

N
H
O
indigo

max for lycopene is at 474 – in the near blue region of

the spectrum – this is absorbed, the compliment is now
red

max for indigo is at 602 – in the orange region of the

spectrum – this is absorbed, the compliment is now 53
indigo!

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UV Spectroscopy
Visible Spectroscopy
Color
1. General
• In the chemical sciences these are the acid-base indicators
used for the various pH ranges:

• Remember the effects of pH on aromatic substituents

Methyl Orange

CH3 CH3
H
O3S N N N O3S N N N
CH3 CH3

Yellow, pH > 4.4 Red, pH < 3.2

54

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