Electronic Spectroscopy

Ultraviolet and visible

 Introduction
• Spectroscopy is the tool for study of atomic & molecular structure.
• It deals with interaction of electronic radiation with matter
involving the measurement & interpretation of the extension of
absorption or emission of electromagnetic radiation by molecule.
• Most important consequence of such interaction is the energy is
absorbed or emitted by the matter in discrete amount called as
quanta.
• UV radiation starts at blue end of visible light(4000Å)
& ends at 2000A.
• It divided into two spectral region-
 Near UV region- 2000Å-4000Å.
 Far or vacuum UV region- below 2000Å.
• UV-spectroscopy involved with electronic excitation.
• Absorption And Emission spectra:-
 Spectroscopy mainly concerned with interaction of
electromagnetic radiation with matter.
 After interaction they may vary in intensity of EMR with
frequency.
 Instrument which record this variation in intensity known as
spectrophotometer
 Two way in which interaction may observed-

Sample itself emits radiation Called as emission spectra

Absorbs radiation from Called as absorption spectra

continuous source
 Terminology
• Wavelength(λ):-
 distance between two successive maxima of one
electromagnetic wave. express in Angstron units or (mu)
• Frequency(ν):-
 Number of wavelength passing through a given point. per sec.
 Unit:- Hertz or cycles per second
• Wave number:-
 Number of waves per centimeter in vacuum.
 Reciprocal of wavelength, express as per (cm).
• relation between frequency, velocity & wave number

ν=(1/λ)c=(c/v)λ=(v/c)
 Electro Magnetic Spectrum

Color Wave
length
109 107 105 103 101 10-1 10-3 10-5 10-7 10-9 10-11

(nm)
violet 400-435
gamma

X-rays
indigo 435-480
500 Violet, indigo, blue
Ultra Violet
600 Green, yellow blue 480-500
700 Orange, red
Infra red green 500-560
microwave
yellow 560-595
Radio
waves orange 595-610

red 610-750
• What compounds show UV spectra?
• Generally think of any unsaturated
compounds as good candidates. Conjugated
double bonds are strong absorbers
• Just heteroatoms are not enough but C=O are
reliable
 Transitions
• s->s*
– UV photon required, high energy
• Methane at 125 nm
• Ethane at 135 nm
• n-> s*
– Saturated compounds with unshared e-
• Absorption between 150 nm to 250 nm
• e between 100 and 3000 L cm-1 mol-1
• Shifts to shorter wavelengths with polar solvents
– Minimum accessibility
– Halogens, N, O, S
• n->p*, p->p*
– Organic compounds, wavelengths 200 to 700 nm
– Requires unsaturated groups
• n->p* low e (10 to 100)
– Shorter wavelengths
• p->p* higher e (1000 to 10000)
Electron transitions
 The UV Absorption process
   * and   * transitions: high-energy, accessible in
vacuum UV (max <150 nm). Not usually observed in
molecular UV-Vis.

 n  * and   * transitions: non-bonding electrons

(lone pairs), wavelength (max) in the 150-250 nm region.

 n  * and   * transitions: most common transitions

observed in organic molecular UV-Vis, observed in
compounds with lone pairs and multiple bonds with max =
200-600 nm.
 An Electronic Spectrum
Make solution of
concentration low
enough that A≤ 1
(Ensures Linear Beer’s law
behavior)
1.0
maxwith certain extinction  Even though a dual beam
goes through a solvent
UV Visible blank, choose solvents
that are UV transparent.
Can extract the  value if
conc. (M) and b (cm) are
known
UV bands are much
Absorbance

broader than the

photonic transition event.
This is because vibration
levels are superimposed
on UV.

0.0
200 400 800

Wavelength, , generally in nanometers (nm)

Chromophore system λ max

Amine -NH2 195 nm

Bromine -Br 208 nm
Iodide -I 260 nm
Thioketone -C=S 205 nm
Ester -RCOOR’ 205 nm
Aldehyde R-CHO 210 nm
Carboxylic acid R-COOH 200-210 nm
Nitro -NO2 210 nm
Nitrite -ONO 220-230 nm
Azo -N=N 285-400 nm
Conjugated olefins (-C=C-)2 210-230 nm
(-C=C-)3 260 nm

(-C=C-)5 330 nm

Benzene 198, 255

Naphthalene 220, 275, 314
 Problems
Which of the following compounds do you expect to absorb ultraviolet radiation? Why or why
not?
i) Acetaldehyde ii) Benzene iii) Cyclohexane iv)Ethanol v) Heptane vi) Aniline vii)
Butadiene
i) Acetaldehyde: It shows absorption at around 255nm because
of low energy n-ᴨ* transition.
ii) Benzene. It absorbs at about 255nm due to ᴨ-ᴨ*
iii) Cyclohexane. It does not absorb in the UV region as it involve
only σ-σ* transition.
iv) Ethanol: It does not absorb UV region because it involves only σ-σ* transition and
n- σ*
v) Heptane: no absorption due to σ-σ* transition.
vi) Aniline: because of the presence of auxochrome, NH2 group attached to benzene it
absorb at 280 nm
vii) Butadiene: It contains conjugated double bonds it absorbs at 217nm
2. Acetaldehyde is having absorption peaks at 160nm, 180 nm
and 292nm. What type of transition is responsible for each of
these absorption?

3. Which of the following compounds absorb UV radiation?

i) Water ii) heptene iii) chloro hexane iv)n-butyl amine v)
acetone vi) benzoic acid
4. Which of the following transition needs greater energy?
i) n-ᴨ* ii) σ-σ* iii) n- σ* iv) ᴨ- ᴨ* ,
5. Benzene exhibits three bands two intense bands at 180-
200 and a weak band at 260 nm. What type of transition is
responsible for these absorptions.
 Change in position and intensity of
absorption
For isolated chromophore groups such as c=c and c ≡ c, absorption takes
place in far ultraviolet region which cannot be studied. But the position of
absorption maximum and the intensity of absorption can be modified in
different ways by some structural changes or change of solvent as fallows:
i) Bathochromic shift or red shift: It involves the shift of absorption
maximum towards longer wavelength because of the presence of
certain groups such as OH and NH2 called auxochromes or by change
of solvent. For example, decreasing the polarity of solvent causes a
red shift in the n-ᴨ* absorption of carbonyl compounds.
Bathochromic shift is also produced when two or more
chromophores are present in conjugation in a molecule.
For example; Ethylene shows a ᴨ-ᴨ* transition at 170 nm where
as 1,3-butadiene shows at 217 nm.
One of the best ways for identifying the presence of
acidic or basic groups, due to big shifts in  for a
chromophore containing a phenol, carboxylic acid, etc.

Hyperchromic shift

“hypsochromic” shift
“bathochromic” shift
Hypochromic shift

ii) Hypsochromic shift or blue shift. It involves the
shift of absorption maximum towards shorter
wavelength and may be caused by removal of
conjugation in a system or by change of solvent. For
example: In aniline , absorption maximum takes
place at 280 nm because the pair of electrons on
nitrogen atom is in conjugation with the ᴨ bond
system of the benzene ring. In acidic medium it
absorbs at 200 nm because the electron pair is no
longer present and hence conjugation is removed.
iii) Hyperchromic shift: This effect involves an increase in the
intensity of absorption and is usually brought about by
introduction of an auxochrome. For example. introduction of
CH3 group in position 2 of pyridine increases є from 2750-3560
and λmax from 253 nm to 262 nm

iv) Hypochromic shift: This effect involves a decrease in the

intensity of absorption and is brought by groups which are
able to distort the geometry of the molecules. For example
when a methyl group is introduced in position 2 of biphenyl
group hypochromic effect occurs because of distortion caused
by methyl group.
Є max

λmax
 Solvent effects
A most suitable solvent is one that does not itself absorb in
the region under investigation.
A dilute solution of the sample is always prepared for spectral
analysis.
Most commonly used solvent is 95 % ethanol.
Ethanol is cheap and is transparent down to 210 nm.
Commercial ethanol should not be used because it is having
benzene which absorbs strongly in the ultra violet region.
Some other solvents which are transparent above 210nm are
n-hexane, water, methanol, cyclohexane, acetonitrile, diethyl
ether etc.
Hexane and other hydrocarbons can be used because these
are less polar and have least interactions with the molecule
under investigation.
• The n-ᴨ* (less intense) transition moves to shorter
Wavelength by increasing the polarity of the solvent.
In n-ᴨ* transition, the ground state is more polar than
the excited state

• ᴨ-ᴨ* (intense) transition. In such a case, the

absorption band moves to longer wavelength by
increasing the polarity of the solvent. The dipole-
dipole interactions with the solvent molecules lower
the energy of the excited state than that of the
ground state
 Instrumentation
Components of spectrophotometer
 Source
 Monochromator
 Sample compartment
 Detector
 Recorder
 SAMPLE

RADIANT WAVELENGTH
SOLVENT
PHOTO- READOUT
SOURCE SELECTOR DETECTOR

Fig.-block diagram of instrumentation of UV-spectrophotometer

 Single beam spectrophotometer

Littrow Prism
mirror

Detector

☼
Sample
Chopper
UV Instrumentation
 Sources
 Hydrogen or Deuterium Discharge Lamps:
 Commonly used ultraviolet spectrophotometers are
hydrogen and deuterium lamps under very low pressure
(0.2-0.5 torr) and low voltage ( about 40 V dc) conditions.
 Heated cathodes provide the essential function of
maintaining the discharge. At less than 360 nm these
discharge lamps provide a strong continuum that fulfills most
needs in the ultraviolet region.
 With fused silica envelopes, work to about 160 nm is
feasible. At higher wavelengths longer than about 380 nm,
the discharge has lower intensity and emissions lines
superimposed on the continuum, thus restricts its use in the
visible region.
Incandescent filament lamps:
• Measurement above 350 nm and into the near infrared to 2.5
μm are usually made with incandescent filament lamps, which
give a continuous spectrum over the range.
• In these lamps a wire filament, generally tungsten, is heated
to incandescence by an electric current.
• The filament is enclosed in a hermetically sealed bulb of glass
filled with an inert gas or a vacuum. Filament s are usually
coiled to increase their emissivity, efficacy, and mean
luminance. Incandescent lamps are rugged, low cost units
sufficiently bright for nearly all absorption work in the visible
and near ultraviolet regions.
Tungsten- halogen lamps :
These are a special class of incandescent lamps with
iodine added to normal filling gases. The envelope is
fabricated of quartz to tolerate higher lamp operating
temperatures of 3500K. The iodine combines chemically
at the bulb wall with sublimed tungsten. The resulting WI 2
migrates back to the hot filament where it decomposes
and tungsten is redeposited. The cycle is repeated ,
continuously cleaning the bulb. These lamps maintain
more than 90 % of their initial light output throughout
their life.
 The Xenon discharge lamps and mercury arcs are also can
be used

 In all sources, excitation is done by passing electrons

through a gas and these collisions between electrons and
gas molecules may result in electronic, vibrational and
rotational excitation in the gas molecules

 When the pressure of the gas is low, only line spectra are
emitted. But, if the pressure of the gas is high, band
spectra and continuous spectra will be obtained
 Monochromators
 The dispersing element may be a prism or grating.

 The prisms are generally made of, quartz or fused

silica which are transparent throughout the entire
range.

 Glass has the highest resolving power but it is not

transparent to radiations having the wavelength
between 200-300 nm because glass absorbs strongly
in this region.
 Detector
Three common types of detectors are used,
1.Barrier layer cell: It consists of a semiconductor such as
selenium, which is deposited on a strong metal base such as
iron and the surface coated with Ag or Au. This cell is known
as a photovoltaic cell
 Photovoltaic cell is entirely different in design and principle
from the photo emissive cell (photocell).
 Photovoltaic cell operates without the use of a battery.
 The metal base plate like iron or aluminium acts as one
electrode.
 Selenium acts as a barrier between inner and outer surface
 Very thin layer of silver or gold which acts as a second
collector electrode.
 When the radiation is incident upon the surface of selenium,
electrons are generated at the selenium-silver interface. These
electrons are collected by the silver. The accumulation of the
electrons on the surface creates an electric voltage difference
between the silver surface and the basis of the cell. If this cell
is connected to a galvanometer current will flow which will
vary with the intensity of the incident light.
Limitations of photovoltaic cell:
1) Its use is generally limited to the visible region, it has less
sensitivity in the blue region
2) The current output depends upon the wavelength of the
incident light
3) The current produced by it cannot be readily amplified by
conventional electronic circuits because of the low internal
resistance.
4) These cells show fatigue effect since the initial photocurrent
is very high and it attains steady value after a few minutes.
This fatigue effect can be minimized by careful selection of
the optimum level of illumination.
2. Photocell.
• A photocell is a type of resistor. When light strikes the
cell, it allows current to flow more freely.
• It consists of a high-sensitive cathode in the form of a
half-cylinder of metal which is contained in an
evacuated tube.
• The inner surface of cathode is coated with a light
sensitive layer such as cesium or potassium oxide and
silver oxide.
• A metal ring inserted near the Centre of the bulb acts
as anode is also present in the tube which is fixed more
or less along the axis of the tube.
• The photocell is more sensitive than photovaltaic cell
as phototubes have high internal resistances , their
output current can be easily amplified and therefore, these
are employed for measuring low intensities of
3. Photomultiplier tube :
 A photomultiplier tube is a combination of a photodiode
and an electron multiplying amplifier.
 A photomultiplier tube consists of an evacuated tube
which contains one photo cathode and 9-16 electrodes
known as dynodes.
 The surface of each dynode is of Be-Cu, Cs-Sb or similar
material
 Advantages:
 By a photomultiplier tube the overall amplification factor
of about 10 6 can be achieved
 The photomultiplier tube can be carefully shielded from
stray light.
 Recorder
Pen recorder
Oscillograph
 Disadvantages of Single-Beam System

 It measures the total amount of light reaching the

detector, rather than the percentage absorbed. Light
may be lost at reflecting surfaces or may be absorbed
by the solvents used to dissolve the sample.
 The source intensity vary with changes in line voltage.
 Response of the detector varies significantly with the
wavelength of the radiation falling on it.
 Different detectors respond differently at different
wavelengths.
 Advantages of Double Beam Instrument

 It is not necessary to continuously replace the

blank with the sample or to zero adjust at each
wavelength as in single beam units.
 The ratio of the powers of the sample and
reference beams is constantly obtained and used.
 Any error due to variation in the intensity of the
source and fluctuation in the detector is minimized.
 Double beam system lends itself to rapid
scanning over a wide wavelength region and to the
use of a recorder or digital read out.
 Sample cells
The cells that are to contain samples for analysis should
fulfill three main conditions

 They must be uniform in construction, the thickness must be

constant and surfaces facing the incident light must be optically
flat.
 The material of construction should be inert to solvents.
 They must transmit light of the wavelength used.

The most commonly used cells are made up of quartz or fused

silica. These are readily available even in matched pairs where
sample cell is almost identical to the reference cell.
 Sample cell (cuvette)
Spectroscopy requires all materials in the beam path other
than the analyte should be as transparent to the radiation as possible.
The geometries of all components in the system should be such as to
maximize the signal and minimize the scattered light.
The material from which a sample cuvette is fabricated controls the
optical
window that can be used. Some typical materials are:
Optical Glass - 335 - 2500 nm
Special Optical Glass – 320 - 2500 nm
Quartz (Infrared) – 220 - 3800 nm
Quartz (Far-UV) – 170 - 2700 nm
•Keep the cuvette clean.
•Don’t clean with paper products.
•Store dry.
•Don’t get finger prints on them.
•Store carefully and gently
 Applications

• Detection of conjugation.
• Detection of geometrical isomers
H5C6 H H 5C 6 C6H5

H H H
C6H5

trans -stilbenes cis -stilbenes

λmax=294nm λmax=278nm
(ε=24000) (ε=9350)

• Detection of functional groups

• Qualitative analysis
• Detection of impurities
• Quantitative analysis
• Molecular weight determination
• Chemical kinetics
• Tautomeric equilibrium

Λmax =275 nm and

O
ε=16
H3C CH 2COOC 2H5 CH 3C(OH) = CHCOOC 2H5

λmax=244nm and ε=16000

 Visible Spectroscopy
• The wavelength range for visible
spectroscopy lies between 400nm-800nm.

• The visible spectroscopy is also known

as
colorimetry
Theory of Spectrophotometry and Colorimetry

When a light is incident upon a homogeneous

medium, a part of the incident light is reflected, a
part is absorbed by the medium and the remainder
is allowed to transmit as such. If I0 denotes the
incident light, Ir the reflected light, Ia the absorbed
light and It the transmitted light, then one can write,
I0 = I a + I r + I t
If a comparison cell is used, the value of Ir, which is
very small (about 4 %)
I0 = I a + I t
Lambert’s Law. This law can be stated as
follows:
“When a beam of light is allowed to pass through a
transparent medium, the rate of decrease of intensity of
the radiation with the thickness of medium is directly
proportional to the intensity of the light”.
Mathematically, the Lambert’s law may be
stated as follows:
-dI/dt α I
Or -dI/dt = kI -------------------(1)
Where I denotes the intensity of incident light of
wavelength λ, t denotes the thickness of the medium
and k denotes the proportionality factor.
On integrating the eqn.1
ln I0/It =kt
or It =I0 e-kt

Where I0 denotes the intensity of the incident light, It denotes

the intensity of the transmitted light and k is a constant
which depends upon the wavelength and absorbing medium
used. On changing equation from natural logarithms, we get,

It =I0 10-0.4343kt = I0 10-Kt

Where K= k/2.3026 , Called as absorption coefficient which is

defined as “It is the reciprocal of the thickness which is
required to reduce the light to 1/10 of its intensity”
• The ratio It/I0 is termed as transmittance and
log I0/It is termed as the absorbance, A, of
medium.
A= log I0/It = -log T
 Beer’s law.
Lambert’s law shows that there exists a logarithmic relationship
between the transmittance and the length of the optical path through
the sample. Beer observed that a similar relationship holds between
transmittance and the concentration of a solution. i.e.,the intensity of
a beam of monochromatic light decreases exponentially with the
increase in concentration of the absorbing substance arithmetically.

Thus eqn., becomes as

It=I0 e-k’c -------------------------(2)

= I0 10-0.4343k’c = I010-K’c
On combining eqs. 1 and 2 called Beer-Lambert’s law
It=I0 10-act or log (I0/It) = act
• The molar absorption coefficient, molar extinction
coefficient, or molar absorptivity, is a measurement of how
strongly a chemical species absorbs light at a given
wavelength. It is an intrinsic property of the species; the
actual absorbance, A, of a sample is dependent on the
pathlength, ℓ, and the concentration, c, of the species via the
Beer–Lambert law, .
• The SI units for ε are m2/mol, but in practice, they are usually
taken as M−1 cm−1 or L mol−1 cm−1.

• designating as an absorption of light when passing through

the one mole of solute per liter of solution having unit length
 Deviations from Beer’s Law

 From Beer’s law it follows that if we plot

absorbance A, against concentration, a straight line
passing through the origin should be obtained.
 But there is usually a deviation from a linear
relationship.
 Deviation from the law are reported positive or
negative according to whether the resultant curve is
concave upwards or concave downwards
There are three types of deviations for Beer’s law :
1.Chemical deviations:
A. Presence of foreign substance whose ions do not
chemically react with the colored compounds may
affect light absorption and may also alter the value of
the extinction coefficient.
B. If the coloured solute ionises, dissociates or
associates in solution. For example, benzyl alcohol
in chloroform exists in a polymeric equilibrium:

4 C6H5CH2OH ↔(C6H5CH2OH)4
Dissociation of the polymer increases with dilution. The
monomer absorbs at 2.750-2.765 µ where as the polymer
absorbs at 3.0 µ. Hence absorption at 2.75µ shows negative
deviation whereas at 3.0µ positive deviation

Another example of deviation is the change of colour of

dichromate ion on dilution. The effect can be represented by
the equilibrium as follows:
Cr2O7 2- + H2O ↔ 2CrO4 2- + 2H+
(Orange) (yellow)

450nm 350 nm
C. Deviation may also occur due to the presence
of impurities that fluoresce or absorb at the
absorption wavelength.
D. Beer’s law cannot be applied to suspension
 Instrumental Deviations
A. Polychromatic Radiation:
Beer’s law requires monochromatic radiation because
absorptivity is a constant at a single wavelength and varies with a
change in wavelength. The possibility of error due to the
practical impossibility of obtaining monochromatic radiation
may be minimized by the selection of a spectral region where
the change in absorptivity with a change in wavelength is very
small. This dictates a wavelength selection from a broad band
rather than from a sharply rising or sharply falling section of the
absorption curve.
The energy which passes through the sample and activates the
detector is a function of the weighted average of the
absorptivities of all the wavelengths which are transmitted by
the monochromator system. At the top (point A) of the
absorption band, the absorptivities at λ1 and λ 2 are close to
that of the desired wavelength (λmax). Minimum error will rise
from the use of this wavelength. On the sharply rising (point B)
or falling (point C) positions the weighted average yields low
results, these low results becoming more and more apparent at
increasing concentrations. In summary, because of the
numerous possibilities of an error arising from deviations it is
necessary in practice to prepare a calibration curve for the
absorbance- concentration relationship at the chosen
wavelength. Beer’s law should be verified in each application
on each instrument. It should not be assumed.
Stray light: Stray light is defined as radiation from the
instrument that is outside the nominal wavelength band
chosen for the determination. This stray radiation often is
the result of scattering and refection of the surfaces of
gratings, lenses or mirrors, filters and windows. When
measurements are made in the presence of stray light, the
observed absorbance is given by
A’ = log[(P0 + Ps)/ (P + Ps)]
Where Ps is the radiant power of the stray light.
The deviation due to stray light are most significant at high
absorbance.
Mismatched Cells.
If the cells holding the analyte and blank solutions are
not of equal path length and are not equivalent in optical
characteristics, an intercept will occur in the calibration
curve and A = εbc + k
Will be the actual equation. This error can be avoided by
using either carefully matched cells or a linear regression
procedure to calculate both slope and the intercept of
the calibration curve. Another way to avoid the
mismatched cell problem with single beam instruments
is to use only one cell for the blank and sample by
placing in the same position after careful cleaning.
 Problems
1. The absorption spectrum for titanium peroxide complex ion in perchloric
acid showed a maximum of 400nm. The absorbance of a 32.0 µg/ml
solution of titanium gave an absorbance of 0.456. An unknown solution
treated in an identical fashion gave an absorbance of 0.501. Assuming
identical cells, find out the concentration of the unknown.
As= єscsts
Ax = єxcxtx
As/ Ax = єscsts/ єxcxtx
Since the absorptivities of the standard and unknown solutions are the
same and the cell depths ts and tx are equal, the equation can be written as:
As/ Ax = cs/ cx
Cx = ?
= 32.0 x (0.501/ 0.456) = 35.2 µg/ml