Bonga university

Instrumental Analysis II (Chem.2052)

Chapter 5
Uv/visible spectroscopy 2012 E.C

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 5.1. Introduction
 For a long time, experimental work made use of the human
eye as the detector to measure the hue and intensity of colors
in solutions.
However, even the best analyst can have difficulty comparing
the intensity of two colors with slightly different hues, and there
are of course people who are color-blind and cannot see certain
colors
 Thus, instruments like UV-Vis have been developed to perform
these measurements more accurately and reliably than the
human eye.
 Most of the organic compounds are transparent in the portion of
electromagnetic spectrum.
 UV-Vis spectroscopy involves absorption of UV-Vis light by
molecules but not by atoms.
 The transition is electronic, therefore it is electronic
spectroscopy.
UV-Vis Spectroscopy (frequently called Electronic
Spectroscopy), is the spectroscopy which utilizes the
UV and Vis range of electromagnetic radiation.
 The UV-Vis wavelength range is commonly between
190 – 800 nm.
 Table 5.1. Ranges of the electromagnetic radiation
used in electronic spectroscopy.

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Radiation  *

nm cm-1
o

VIS 400-750 4000-7500 25000-13000

near UV 200-400 2000-4000 50000-25000

Far UV(vacuum UV) < 200 < 2000 >50000

Ranges of the electromagnetic radiation used in electronic

spectroscopy

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5.2. Basic Principles
Absorption of radiation in the visible and uv
regions of the electromagnetic spectrum results in
electronic transitions between molecular orbitals.
Electron is promoted from an occupied molecular
orbital to unoccupied orbitals. The unoccupied
orbital’s are in higher energy than the occupied
orbitals.
During electronic transition (excitation), there are
also absorption processes that cause rotational and
vibrational excitations.
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 Cont…
Rotational λ > Vibrational λ > Electronic λ
Energy of electronic > Energy of vibrational > Energy
of rotational
 Thus, the rotational and vibrational levels are
superimposed on the electronic levels leading to
abroad band centered near the λ of the major
transitions unlike the band of AAS which is sharp.
 Example: the absorption spectrum of benzene in
solution and gas phase
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samples run in gas
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Samples run in solution
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 Cont….
Such vibrational and rotational fine structure lines are not
usually observed for samples run in solution because of
physical interactions between solute and solvent molecules
which cause collisional broadening of the lines.
 The resulting overlapping bands coalesce to
give one or more broad band-envelopes.
Though the resolution of commercial instrumentation is not
high enough to separate these lines, the fine structure due to
vibrational and rotational sublevels are never observed in routine
UV/VIS spectra.

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 Cont….
Characteristics Features of UV-Vis Absorption
Spectrum
is usually recorded as plot of absorption versus
λ or ε versus λ at which maximum absorption is
observed.
 An absorption band is characterized by its
shape, that
is, by its width and intensity.
 The shape of the band is determined primarily by the
vibrational energy level spacing and the intensity of
each vibrational transition.
 The intensity distribution is related to the probability
of the transition to a given vibrational sublevel.
 The position of the peak is characterized by λmax
 The intensity of the peak is characterized by ε 10
5.2.2. The Magnitude of Molar Absorptivities
Empirically, molar absorptivities (ε values) that
range from 0 up to a maximum on the order of 105 L
mol-1 cm-1 are observed in UV-visible molecular
absorption spectrometry.
 Commonly ε 104–105L mol-1 cm-1 for an
allowed transition and is on the order of 10–100
for a forbidden transition.
The magnitude of the absorptivity is an indication of
the probability of the electronic transition.
High values of ε give rise to strong absorption of light
at the specified wavelength; low values of ε result in weak
absorption of light.
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5.2.3. Types of Electronic Transition
Molecules are composed of atoms that are held together
by sharing electrons to form chemical bonds.
When the molecule becomes excited, an outer
shell (valence) electron moves to an orbital of higher
energy.
 The process of moving electrons to higher energy
states is called electronic excitation.
 For radiation to cause electronic
excitation, it must
be in the visible or UV region of the
 Generally, there are three types of electronic
transitions – namely:
1. Transition involving
(in most organic molecules)
2. Transition involving d and f electrons (most
transition metal ions, lanthanide and actinide
series absorb UV or visible light due to d and f
electrons transition respectively) and
3. Transition involving charge transfer
electrons
(involving the transfer of an electron between two orbitals one
of which is associated predominantly with the ligand and the other
with the metal).
Absorbing Species Containing
 Three distinct types of electrons are involved in valence
electron transitions in molecules.
1. The electrons involved in single bonds
2. The electrons involved in double and triple
(unsaturated) bonds.
3. Electrons that are not involved in bonding between
atoms
These are called n electrons, for nonbonding electrons. In
saturated the outer shell electrons of carbon and hydrogen are all
involved in bonding; hence these compounds do not have any n
electrons.
 Organic compounds containing nitrogen, oxygen, sulfur, or
halogens, however, frequently contain electrons that are
nonbonding (Fig. 5.7).
 Because n electrons are usually excited by UV or visible radiation,
many compounds that contain n electrons absorb UV/VIS
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 Electronic Transition of Organic Molecules
 According to molecular orbital theory, the
interaction of atomic orbitals leads to the formation
of bonding and antibonding molecular orbitals.
 Depending on the nature of the overlapping atomic
orbitals, molecular orbitals may be of the and
(bonding) or * and * (antibonding) orbitals
and n (non bonding orbital).
The type electron densities are
concentrated along the internuclear axis, and the
type, the electron density is concentrated on
either side of the internuclear axis.
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When molecules are electronically excited,
an electron moves from the highest occupied
molecular orbital (HOMO - n) to the lowest
unoccupied orbital (LUMO- *), which is usually
an antibonding orbital.
 Electrons in  bonds are excited to antibonding
*orbitals, and n electrons are excited to either

* or * orbitals.
i. * transition

E large and occurs in vacuum UV region (λ<150 nm), ε =10 -

10,000 L/mol.cm. It is not commonly used for analytical purpose.
This is because the component of air like N, O, which absorbs
energy, so to avoid them we need to expensive energy.
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This is the reason for saturated hydrocarbons that only contain this type
of bonding being transparent in the near UV. i.e. saturated hydrocarbons
contain only single bond (they don’t contain lone pair of electrons or
multiple bonds).
Example:
1.methane contains only single C-H bonds exhibits a at λmax
=125nm,
2.Ethane has also an absorption peak at 135 nm but here electrons of
the C-C appeared to be involved. Because the strength of the C-C is
less than that of the C-H bond, less energy is required for excitation:
thus the absorption peak occurs at a longer wavelength.

Note: as the length of the hydrocarbon increases the λ

will also increases.
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 ii. * transition
 Saturated compound containing atoms with unshared electron pairs
(nonbonding) are capable of * transition
 In general these transition requires less energy than the *
transition.
 They can brought about by radiation in the region between 150 and
250 nm, with most absorption peaks below 200 nm (leads to
moderate intensity located around 180 nm for alcohol, near. 190nm
for ethers or halogen derivatives and in the region of 220nm for
amines.
Examples:
1.methanol: λmax = 183nm (ε = 50) ether: λmax = 190nm (ε = 2000)
ethylamine:
λmax =210nm (ε =800); 1-chlorobutane: λmax =179nm.
Transition depends on the electron negativity of hetroatoms and the nature
of the solvents.
simple,
 aliphatic
Examples: examples
Alcohols, of these
ethers, compounds
amines and sulfurthe n  * is
compounds – thethe
in most often
cases of
observed transition; like the alkane  * it is most often at shorter than
200 nm
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Example: Absorption due * transition
to Compound λmax εmax
H 2O 167 1480
CH3OH 184 150
CH3Cl 173 200
CH3I 258 365
(CH3)2S 229 140
(CH3)2O 184 2520
CH3NH2 215 600
(CH3)3N 227 900
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The molar absorptivity associated with these type of
transition are low to intermediate in magnitude and usually
ranges between 100 and 300 L mol-1.cm-1

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iii. n * transition
 This transition of low intensity results from the
passage of an n electron (engaged in a non-bonding
MO) to an anti-bonding * orbital.
This transition is usually observed in molecules
containing a hetero atom carrying lone electron pairs as
part of an unsaturated system.
 The best known is that corresponding to the carbonyl
band, easily observed at around 270 to 295 nm. The
molar absorption coefficient for this band is weak.
Example: ethanal: λ = 293nm (ε=12, with ethanol as
solvent).
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Organic molecules that contain atoms with non
bonded electrons should be able to absorb
UV/VIS radiation.
Such atoms include nitrogen, oxygen, sulfur, and
the halogen atoms, especially Br and I. Table 5.2
presents some typical organic functional groups that
serve as chromophores.
Table 5.2. Organic Functional Groups that can
Absorb UV/VIS Radiation
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Table 5.2

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 The molar absorptivity for peaks associated to
the
n, * state are generally low and ordinarily
ranges
from 10 to 100 L mol-1.cm-1
 Peaks associated with n * transitions
are generally shifted to shorter λ (a blue shift
with increasing polarity of the solvent.
iv.  * transition
 This transition to occur, a molecule must possess
a chromophore with an unsaturated bond, such
as
C=C, C C, C=O, C=N, N=O,N=N and so on.
Compounds with these types of chromophores include
alkenes, amides, ketones, carboxylic acids, and oximes,
 Exhibited by compounds containing multiple bonds like alkene, alkyne etc.
 Is affected by conjugation, as conjugation increases the wavelength also increases.
 Values of ε for  * transition fall in the range between 1000 and 10,000 L mol-
1.cm-1

 Usually but not always, a red shift is observed for  * transition with
increasing polarity of the solvent.
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Table 5.3 lists types of organic compounds and the
wavelengths of their absorption maximum,

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 In most organic compounds the bonding and non-bonding
orbitals are filled and the antibonding orbitals are vacant.
 The lowest energy and therefore the longest wavelength
transitions are from non-bonding orbitals to antibonding 
orbitals, i.e. n*. These give rise to bands in the near UV
and visible regions.
 Other allowed transitions in order of increasing energy
(shorter wavelength) are n σ* and *, which have
comparable energies, and σσ*. The latter occur in the far UV
or vacuum region below 200 nm and are of little use analytically.
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Of these transitions, the most important are the n* and *,
because they involve functional groups that are characteristic of
the analyte and wavelengths that are easily accessible.

5.3. Absorption characteristics of some chromopores

Definition of some terms:
Chromophore:
 is a group of atoms (part of a molecule) that gives rise to an
electronic absorption.
 an identifiable part of a molecule responsible for any
spectral features arising from electronic transition.
are may be functional groups (like double bond, hydroxyl group,
carbonyl...) or single atom within molecule or group (like methyl).
 The detection of chromphore permits to deduce presence of
structural element or fragment in the molecule from which the
molecular structure may be built up.

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 The max and  values for some typical
chromophores are given in tables following

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33
 Cont…
Auxochrome (Auxilary chromophores):
is a substituent that contains unshared
(nonbonding) electron pairs, such as OH, NH, and
halogens.
 Are groups with very little UV/Vis absorption by
themselves, but which often have significant effects
on the absorption (both max and ) of chromophore
to which they are attached.
 Generally, groups containing heteroatoms with one
or more lone pairs like –OH, –Cl, –OR and –NR2, -
halogens are auxochromes.
 In general, substituents may have any of
four effects on a chromophore:
1. Bathochromic shift (red shift) – a shift to longer ; lower
energy
2. Hypsochromic shift (blue shift) – shift to shorter  higher
energy
3. Hyperchromic effect – an increase in intensity()
4. Hypochromic effect – a decrease in intensity()

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 Factors affecting the position intensity of the
absorption band
 The positions and intensities of the absorption
bands are sensitive to:
1. substituents close to the chromophore,
2. conjugation with other chromophores, and
3. Solvent effects.
1.Effect of Auxochrome (substituents close to the
chromophore)
In general, auxochromic substitution of
chromophores causes bathochromic shifts and increases
in intensity for
* transitions, and hypsochromic or blue shifts (to
shorter wavelengths) for n * transitions.
 Cont….
 The shifts are explainable in terms of mesomeric
(resonance) effects caused by interaction of lone
pair electrons associated with such auxochromes
as —OH, —Cl, —NH2 with the  system of the
chromophore.
 This leads to increases in the energies of  and
* orbitals, the  being raised by more than the
*, but leaves the energy of the non-bonding
orbital unchanged.
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Contd
.

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2. Conjugation Effects
 Increase the amount of conjugated double bonds always
in
shift absorption maxima towards longer wavelength
significantly
(bathochromic) and usually towards stronger intensity relative to an
isolated chromophore.
The shift can be explained in terms of interaction or delocalization
of the  and * orbitals of each chromophore to produce new
orbitals in which the highest  orbital and the lowest * orbital are
closer in energy.
Figure 5.9. shows the conjugation of two ethylene chromophores to
form 1,3-butadiene. The * transition in ethylene occurs at 165 nm
with an εmax value of 1500 whereas in 1,3-butadiene the values are 217
nm and 2100 respectively.
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• Red shift of max with increasing conjugation
CH2=CHCH2CH2CH=CH2 max = 185 nm
CH2=CHCH=CH2 max = 217 nm
CH2=CH-CH=CH-CH=CH2 (trans) max = 263 nm
•Red shift of max with number of rings
• Benzene max = 204 nm
•Naphthalene max = 286 nm 40
 If two unlike chromophores are conjugated and one
group has nonbonding electrons, the n * transition
is also shifted bathochromically because the energy of
the antibonding orbital is lowered.
 Thus the weak n * band in a saturated carbonyl
compound is shifted from below 300 nm to above 300
nm with an increase in .
 Conjugation of additional chromophoric groups
moves λmax progressively towards the visible region
and increases ε.
For example:
 tetradecahexaene (six double bonds) absorbs at the blue end
of the visible region and appears yellow whilst with further
conjugation, as in the carotenes (ten or more double bonds),
the compound may appear orange, red, purple, or even
black.
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 The presence of an auxochrome that extends
the double bond causes a red shift.
Example: H2C and CH2
λmax λmax
 They have the
< same functional group why the
difference? This is because of conjugation, the larger the
conjugation, the larger the λ
Absorption by aromatic compounds
 Absorption of aromatic hydrocarbons are characterized by
three sets of bands. Benzene can be regarded as a special case
of a conjugated triene. It shows λ* at 184, 204 and 254
nm.
 The last one is said to the normal absorption band,
which shifts bathochromically and intensifies on
chromophoric substitution.
 Of more importance is that substitution produces a new and
intense band between 200 and 300 nm which arises from a
* transition in the extended conjugated system.
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 254 nm 275 nm 280
 Asnm
the number of the ring increases, the absorption λ also
increases why? This is b/c of conjugation.
Example: Substituent effect on benezene absorption maximum
Substituent of benzene  log Substituent of benzene  log
max max

none 204 7900 3.9 -NH2 230 8000 3.9

256 200 2.3 281 160 3.2

-CH3 208 8000 3.9 -OH 211 6300 3.8

261 300 2.5 270 1500 3.2

-Cl 216 8000 3.9 -NO2 251 9000 4.0

265 1500 2.4 280 1000 3.0
330 130 2.1

-OCH3 220 8000 3.9 -CH=CH2 244 12000 4.1

272 1500 3.2 282 1600 3.2 43
Table 5.5. Absorption characteristics of some
conjugated chromophores and chromophorically
substituted benzenes

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3. Solvent Effects
 Solvent polarity changes max since polarity changes with the
movement of electron from one orbital to another.
 Absorption bands arising from n * transitions suffer
hypsochromic shifts on increasing the solvent polarity, whilst
those of * transitions are shifted bathochromically. i.e.
 Polar solvents cause a blue shift (lower λ) due to salvation of
the nonbonding electrons lowering the n orbitals But they
cause a red shift for stabilizing the * orbitals than do the  (
polar solvents will stabilize both  and  but it stabilizes *
stronger than ). On the other hand, the reverse is true if the
solvent is non polar.
Explanations:
 The energy of the non-bonding orbital is lowered by
hydrogen
bonding in the more polar solvent thus increasing the energy
of the
n to * transition, but the energy of the * orbital is
decreased relative to the  orbital.
 Solvent can also induces significant changes in the intensity
of peaks.
Hyperchromic – Increase in absorption peak
Hypochromic – Decrease in absorption
peak
Example:
1. For acetone, n to * transition occurs at 279 nm in n-
hexane, 270 nm in ethanol and at 265 nm in water
Solvent
2. Absorption characterstics of 2 – methylpyridine
λmax εmax
Hexane 260 2000
Chloroform 263 4500
Ethanol 260 4000
Water 260 4000
Ethanol – HCl (1:1) 262 5200

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 Woodward’s Rules or the Woodward–Fieser Rules
 The rule is governed by additive effect
of auxochrome substituent.
 The incremental contribution of substituents is
added to this base value from the group tables:
Table 5.6 Empirical Rules for Calculating the
Absorption

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48
The parent λ = 253 nm λ = 214 nm Cyclic
absorption λ = 217 nm
Acyclic

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Examples

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 INSTRUMENTATION FOR UV-VISILE
SPECTROSCOPY
Spectrometric instruments have a common set of
general features. Often, one technique is distinguished
from another by differences in these features. Here we
look at specific features for the UV/Visible experiment.
1.Sources: D2 lamp, W filament (halogen lamp), and Xe
arc lamp.
2.Wavelength Selectors: Filters and Monochromators.
3.Sample Containers: Fused silica, quartz, and glass.
4.Detectors: Phototube, PMT, photodiode, photodiode
array, CCD array
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 Application
 Have Both qualitative and quantitative analysis.
 Used In analytical chemistry for the quantitative determination
of
different analytes, such highly conjugated organic compounds,
and biological macromolecules.
Identification is done by comparing the absorption spectrum with the
spectra of known compounds.
UV absorption spectroscopy is generally used for
characterizing aromatic compounds and aromatic olefins etc.
THANK
YOU!

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