UV-Visible Spectroscopy

INTRODUCTION:

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).

PRINCIPLE:

The principle involved is the transitions of electrons from lower energy level to higher energy level. 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.
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. It is due to addition of auxochrome.

Hypochromic effect or shift: It leads to decrease in absorption intensity. It occurs due to removal of
auxochrome or conjugation.

INSTRUMENTATION:

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

 1. 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.
1. Filter or Monochromator: These are devices used to selectively allow the radiations to pass through
the sample. 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
2. 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.
3. 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.
4. Amplification and recording unit: The electrical signals are amplified and output generation is on
computer monitor or graphical print out.

Working: 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 (C 1) 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. It is done by measuring the change in concentration of reactant or product with time provided one
of the reactant or product exhibit absorption in UV-Vis region. As absorbance is directly proportional to
concentration, the same can be used to understand the rate of reaction.

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

PRINCIPLE: 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, bond length remaining constant. These vibrations can
occur within the same plane or out of plane. There are four types of bend: Rocking, Scissoring, Wagging and
Twisting.
Rocking: It involves the movement of atoms in same direction

Scissoring: In this, two atoms joined to the central atom move toward and away from each other with change in
bond angle.

Wagging: The atoms move up and below the plane with respect to central aom.

Twisting: It involves the movement of one atom up and other atom down the plane with respect to the central
atom.

INSTRUMENTATION:

Components of IR Spectophotometer:

2. 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.
3. In double beam operation, a beam splitter separates the incident beam in two; half goes to the sample,
and half to a reference.
4. A slit selects the collection of wavelengths that shine through the sample at any given time.
5. 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
6. 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.
 7. 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.
8. 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. (1300-
900 cm-1). This range is the fingerprint of the compound. If two samples gives similar pattern of bands
in this region, this can be considered as a proof that the two samples are of the same compound. No two
different compounds give same IR spectra. It thus helps in identification of the unknown substance.

2. Determination of quality of the substance: There occurs appearance of an extra band in case any
impurity is present in a compound. For example, the small amount of ketone in ahydrocarbon is detected
by the appearance of a band near 1720 cm-1, which is characteristic frequency of the carbonyl group.

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.