UV-Visible Spectroscopy

Spectroscopy: It is the study of the interaction of the EMR with matter. It is the analysis of the
EMR scattered, absorbed or emitted by molecules.

UV-Visible Spectroscopy: When ultra violet and visible range of electromagnetic radiations
interacts with matter it is called UV-Visible Spectroscopy. The wavelength in this spectroscopy
ranges from 100nm to 700 nm. It is also called Electronic spectroscopy because it involves
transitions of electrons from lower energy level to higher energy level. Electronic transitions are
accompanied by vibrational and rotational transitions which lead to the appearance of bands. (That
is why band are broad instead of fine lines).

UV- visible spectroscopy is used to analyze the organic compounds. These compounds are
characterized by the presence of three types of electrons (, n and  electrons). The principle
involved is the transitions of these electrons from bonding molecular orbitals to antibonding
molecular orbitals on absorption of energy.

Following types of transitions are possible on absorption of electromagnetic radiations (UV-Vis)

 → * : An electron in a bonding  orbital is excited to the corresponding antibonding orbital.

The energy required is large. For example, methane (which has only C-H bonds, and can only
undergo  → * transitions) shows an absorbance maximum at 125 nm. Absorption maxima
due to  → * transitions are not seen in typical UV-Vis. spectra (200 - 700 nm)

n → * and  → *: Most absorption spectroscopy of organic compounds is based on transitions

of n or  electrons to the * excited state. This is because the absorption peaks for these transitions
fall in an experimentally convenient region of the spectrum (200 - 700 nm). These transitions need
an unsaturated group in the molecule to provide the  electrons (alkenes, conjugated dienes,
ketones ).
n → *: Saturated compounds containing atoms with lone pairs (non-bonding electrons) are
capable of n→ * transitions. These transitions usually need less energy
than  → * transitions. They can be initiated by light whose wavelength is in the range 150 -
250 nm. The number of organic functional groups with n→ * peaks in the UV region is small.

Note:

Chromophore: Chromophore in Greek means colour carrier. Originally it was any group
responsible for imparting colour to the compounds. It is also defined as any isolated covalently
bonded group showing absorption in uv visible region. Examples are Ethylenes, Acetylenes,
Carbonyls, Nitriles, Azo group

Auxochrome: Any color enhancing group is called auxochrome . It by itself does not act as an
chromophore. It is a saturated group with non-bonding electron when attached to chromophore
alters both wavelengths as well as intensity of absorption Examples are Hydroxy, amines, alkoxy,
thiols.

Intensity Shifts

Bathochromic shift or effect :(Red shift) Shift of lambda max (λmax) to longer wavelength or less
towards energy is called bathochromic shift or red shift. This is due to substitution or solvent
effect or due to presence of auxochrome. The group which deepens the color of chromophore is
called bathochromic group. e.g. Primary,secondary and tertiary amino groups

Hypsochromic shift or effect :(Blue shift) Shift of lambda max (λmax) to shorter wavelength and
higher energy is called hypsochromic or blue shift. e.g solvent effect, removal of auxochrome or
removal of conjugation.

Hyperchromic effect or shift: It leads to increase in absorption intensity

Hypochromic effect or shift: It leads to decrease in absorption intensity

 Types of Spectrophotometer
Spectrophotometer or spectrometer is the instrument used for the study of interactions of matter
with the EMR (absorption measurements).

a) Single beam Spectrophotometer (b) Double beam Spectrophotometer

Working of Spectrophotometer
Source: Hydrogen discharge lamps, deuterium lamps, xenon discharge lamps are the most
common source used in UV spectrometer. Tungsten lamp is used for visible radiations.

Filter or Monochromator: These are devices used to selectively allow the radiations to pass
through the sample.

Sample holder: A quartz cell is used to keep the sample and should be uniform in construction
and inert to the solvent. In case of colorimetery, a test tube can be used for sample holding.

Detector: This unit collects, detects and converts radiant energy into measurable electrical signals.
Photomultiplier tubes, barrier layer cell detectors are most commonly used in spectrophotometer.
Amplification and recording unit: The electrical signals are amplified and output generation is
on computer monitor or graphical print out.

The radiations coming from the source is allowed to passes through the monochromator or filter
via a mirror system. The radiation of the narrow beam divides into two beams of equal intensities,
one passing through the sample and other through the reference cell. The detector receives the
emerging signals and converts it into the graphical form. The absorbance or transmittance is
recorded as a function of wavelength.

Applications of Electronic Spectroscopy

1. Characterization of aromatic compounds and conjugated dienes or olefins: The organic

compounds are characterized by specific λ max.
2. Detection of impurities: The absorption spectrum of the compound is noted using
spectrophotometer. If any overlapping of the spectrum is obtain that suggests presence of
impurity. For example, benzene is common impurity in cyclohexane and its presence can
be easily detected by absorption at 255 nm.
3. Purification of compounds: The process of purification is continued till the compound
being purified stops showing bands due to impurity.
4. Determination of unknown concentration: As absorbance is directly proportional to the
concentration of the compound (Beer lambert’s Law), the unknown concentration can be
determined using known sample. Absorbance of known solution (A1 standard) and
unknown solution (A2) is measured with the help of spectrometer. Knowing the
concentration of standard solution (C1) the concentration of unknown solution (C2) can be
determined by employing,.
A1/C1 =A2/C2
5. Determination of molecular weight by making suitable derivative which absorbs in the UV-
Visible region.
6. Kinetics of chemical reaction can be studied by measuring the change in concentration of
reactants with time.
 Infra Red Spectroscopy
Introduction: It is also called IR Spectroscopy or Vibrational spectroscopy. It involves transition
between vibrational energy levels. Infrared (IR) electromagnetic radiation causes vibrations in
molecules (wavelengths of 2500-15,000 nm or 2.5 – 15 mm). Any change in shape of the
molecule- stretching of bonds, bending of bonds, or internal rotation around single bonds give rise
to IR spectra.

Region Wavelength range (mm) Wavenumber range (cm-1)

Near 0.78 - 2.5 12800 - 4000

Middle 2.5 - 50 4000 - 200

Far 50 -1000 200 - 10

Theory:

For a molecule to absorb IR, the vibrations or rotations within a molecule must cause a net change
in the dipole moment of the molecule. The alternating electrical field of the radiation interacts with
changes in the dipole moment of the molecule. If the frequency of the radiation matches the
vibrational frequency of the molecule then radiation will be absorbed, causing a change in the
amplitude of molecular vibration. The positions of atoms in a molecule are not fixed; they are
subject to a number of different vibrations.

Theory:

For a molecule to absorb IR, the vibrations or rotations within a molecule must cause a net change
in the dipole moment of the molecule. The alternating electrical field of the radiation interacts with
changes in the dipole moment of the molecule. If the frequency of the radiation matches the
vibrational frequency of the molecule then radiation will be absorbed, causing a change in the
amplitude of molecular vibration. The positions of atoms in a molecule are not fixed; they are
subject to a number of different vibrations.

Vibrations fall into the two main categories of stretching and bending.

Stretching is the change in inter-atomic distance along bond axis. It can be symmetrical or
unsymmetrical.
Bending refers to change in angle between two bonds. There are four types of bend: Rocking,
Scissoring, Wagging and Twisting.

Instrumentation:

Components of IR Spectophotometer:

1. A source generates light across the spectrum of interest. The main sources are nichrome
wire wound on a ceramic support or Nernst-glower or globar lamp.
2. In double beam operation, a beam splitter separates the incident beam in two; half goes to
the sample, and half to a reference.
 3. A slit selects the collection of wavelengths that shine through the sample at any given time
.
4. Prism or gratings act as monochromator and is used to disperse a broad spectrum of
radiation and provide a continuous calibrated series of electromagnetic energy bands of
determinable wavelength or frequency range
5. The sample absorbs light according to its chemical properties. Gaseous samples are
collected in a gas cell, liquid samples are placed in cavity cell or sand- witch cell while
solid samples are placed on pressed plate of alkali halide.
6. A detector collects the radiation that passes through the sample, and in double-beam
operation, compares its energy to that going through the reference. It converts thermal
energy into electricity. Detectors commonly used are photocells, photoconductive cells,
thermal detectors etc.
7. The detector puts out an electrical signal, which is normally sent directly to an analog
recorder.

Applications of IR Spectroscopy:

1. Identification of unknown substance: This is done by the fingerprint spectra of the

compound.

2. Determination of quality of the substance: There occurs appearance of an extra band in

case any impurity is present in a compound.

3. Identification of the functional group of the compound: As functional groups in an organic

compound have the characteristics absorption pattern in IR hence the presence and absence
of certain compounds can be known.

4. Structure determination: It gives valuable information about molecular symmetry, dipole

moments, bond lengths etc. which helps in structure elucidation.

5. Distinguishing between intra and inter Hydrogen bonding. This is done by taking a series
of IR spectra at different concentrations. As the concentration increases the absorption band
due to intermolecular H-bond increases while due to intra molecular H-bond remains
unchanged.