CH10001 CHEMISTRY

SPECTROSCOPY

Dr. Janeka Gartia

KIIT University
 Syllabus
Introduction

UV-Vis spectroscopy: Beer Lambert’s law, Types of

transition, Concept of auxochrome and chromophore,
factors affecting , Woodward-Fieser rules for calculation of
lmax in diene systems.

IR spectroscopy: Introduction, types of vibration,

selection rules, Hooke’s law, problem, identification of
functional groups like C-C, C=C, -OH, -NH2 and -C=O.

NMR spectroscopy: Basics of NMR spectroscopy,

chemical shift, Shielding and de-shielding effect, Factors
affecting chemical shift,

Structural elucidation of simple compounds

 What is Spectroscopy ?
Spectroscopy is a technique which concerned with the
study of properties, composition and structure of
matter by interaction with electromagnetic radiation

Importance of spectroscopy:
Molecules are of so small dimensions that it is not possible
To see them. Our knowledge of molecular structure is directly
derived from spectroscopy.
Electro-Magnetic Spectrum
 Spectroscopy

• Spectroscopic method of
analysis involves the
measurements of the
intensity and wavelength of
radiation.
• This provides the basis for
sensitive methods of
detection and quantitation.
• Involves the use of
equipment, which may be
simple, or extremely
complex, with design
features involving the latest
technological development.
 Electromagnetic Radiation
Radiation is a form of energy which has both magnetic and electrical
properties, hence called electromagnetic radiation. The electromagnetic
radiation covers a long range of radiations which are described or
characterized by either wavelength or frequency.

Characteristics
1. Produced by the oscillation of Electric charge and magnetic field residing on the
atom, which are mutualy perpendicular to each other and are coplanar.
2. Characterized by their wavelength or frequency or wave number.
3. Energy carried by electromagnetic radiation is directly propertional to its
frequency. Emission or absorption is quantized and each quantum of radiation
is called a photon.
4. All type of radiation travel with the same velocity and no medium is required.
They can travel through vacuum.
5. A group of elecromagnetic radiation can be split up into various components for
analysis.
 Electromagnetic Radiation
Wavelength (l) is defined as the distance between the successive peaks,
which is measured in the unit nanometers (nm).

Wave number (ῡ) is the raciprocal of wave length, which is measured in

the unit cm-1.

Frequency (u) of radiation is defined as the number of successive peaks

passing a given point in 1 second, which is measured in the unit cycles per
sec or Hertz (Hz). 1 Hz = 1 cycle sec-1.

The relationship between these is:

h = 6.626 ´ 10-27 erg sec

c = 2.998 ´ 1010 cm sec-1
 Visible Spectrum
The visible spectrum is the portion of the
electromagnetic spectrum that is visible to the human eye.
Electromagnetic radiation in this range of wavelengths is
called visible light or simply light. A typical human eye will
respond to wavelengths from about 400 to 700 nanometers.
 Origin of spectrum

Consider two energy levels of an atom/molecule. The one

with lower energy is called ground state energy level (Eg) and
the one with higher energy is called excited state energy level (Ee).
 Emission Spectra Absorption Spectra

Produced when atoms release Produced when atoms absorb

energy energy
It is a discontinuous spectrum It is a continuous spectrum
consisting of few wavelengths with consisting of all the wavelengths
distinct spacing between them having dark lines or gaps between
them
The type of photons emitted is The wavelengths of light absorbed
helpful in figuring out the kind of is helpful in figuring out the quantity
elements the substance is made of of substances in the sample.
as each element radiates a different
amount of energy and has a unique
emission level
 Types of Spectra
Emission or absorption spectrum can be of two types.

Atomic spectra
When atoms interact with electromagnetic radiation, the
spectrum obtained is called an atomic spectrum. The
transition of electrons takes place between the electronic
energy levels of the atom. The spectra are obtained as sharp
lines. These lines are characteristic of the atom, hence, line
spectra are also regarded as finger prints of the atoms.

Molecular spectra
Molecular spectroscopy deals with the interaction of
electromagnetic radiations with molecules. In the spectra of
molecules, in addition to electronic levels, transitions also
occur between rotational and vibrational levels.
 Interaction of Radiation with Matter
Radiation is a form of energy which could either be absorbed or
transmitted. The interaction of radiation in spectroscopy involves
transition between the different energy levels of atoms or molecules.

Generally, the absorption or emission of radiation by matter involves the

exchange of energy. Thus, to understand the principle of this exchange,
it is necessary to know the distribution of energy within an atom or
molecule.

The internal energy of a molecule is due to the energy associated with:

• The electrons
• Vibrations between atoms
• The rotation of various groups of atoms within a molecule
The energy levels can be altered by the absorption or emission of
radiant energy. This is because atoms exist only within a limited number
of energy levels. A study of the wavelength or frequency of radiation
absorbed or emitted by an atom or molecule will give information about
its identity. This technique is known as qualitative spectroscopy.
 Types of spectroscopy

Based on the nature of the radiation that is being absorbed

or emitted, there are different types of spectroscopy.

• Ultraviolet/visible spectroscopy
• Infra-red (IR) spectroscopy
• Nuclear Magnetic Resonance (NMR) spectroscopy
• Mass spectroscopy
• Flame Photometry
• Atomic Absorption Spectroscopy
• Raman spectroscopy
• X- ray spectroscopy
• Fluorescence spectroscopy
• Mossbauer spectroscopy
UV-visible Spectroscopy
 It is also known as Electronic Spectroscopy since it
involves the promotion of electrons (s, p, n electrons) from
the ground to the higher energy states.

• It is useful to measure the number of conjugated

double bonds and also aromatic conjugation within
various molecules.

• Distinguishes between conjugated and non-conjugated

systems.

• Distinguishes homoannular and hetero annular

conjugated dienes.

Since the energy levels of a molecule are quantized, the energy

required to bring about the excitation is a fixed quantitiy.
 The UV-Vis spectral region

• This region of the spectrum is conventionally divided into three sub-

domains termed
UV (185–400 nm),
Visible (400–700 nm) and
Near Infrared (NIR) (700–1100 nm).
• Most commercial spectrophotometers cover the spectral range of 185 to
900 nm.
 The origin of the absorptions
• The origin of absorption is the interaction of photons from a source with
ions or molecules of the sample.
• When a molecule absorbs a photon from the UV-Vis region, the
corresponding energy is captured by one (or several) of its outermost
electrons.
• As a consequence there occurs a modification of its electronic energy
Eelec, a component of the total mechanical energy of the molecule along
with its energy of rotation Erot and its energy of vibration Evib.
Types of energy changes
A number of energy states are possible in molecules. The total
energy is the sum of the translational, rotational, vibrational and
electronic energy.

Thus, there are three types of energy changes in a molecule

1. Due to transition of an electron from one orbit to another.
2. Due to rotation of the molecule.
3. Due to vibration of atoms of the molecule relative to one another.
• Rotational energy (E rot): It involves the rotation of molecules about the
centre of gravity or of parts of molecules.

• Vibrational energy (E Vib ): It is associated with stretching, contracting,

or bending of covalent bonds in molecule.

• Electronic energy (E Elec ): It involves changes in the distribution of

electrons by splitting of bonds or the promotion of electrons into higher
energy levels.
 Beer-Lambert’s Law
It states that when a beam of monochromatic radiation is passed through a
solution of an absorbing substance, the rate of decrease in intensity of
incident radiation with thickness of absorbing solution is directly
proportional to to the product of the intensity of incident radiation and
concentration of the solution.

−
e Is called molar absorption coefficient or molar absorptivity or
molar extinction coefficient

If concentration and thickness of absorbing solution becomes unity,

i.e., C = 1 mol/L and x = 1 cm, then
A=e
Thus, molar absorption coefficient (e) can be defind as the absorbance
(A) of an absorbing solution of unit concentration and unit path length.
It depends on the frequency of incident radiation and is the largest
when absorption of radiation takes place to a greater extent.
Its unit is Conc-1 length-1

where, T is the transmittance

 UV-Vis spectrophotometer mechanism

Courtsey: Jasco Inc.

This law, which concerns only that fraction of the light absorbed can
be verified by application of the following conditions:

• the light used must be monochromatic

• the concentrations must be low
• the solution must be neither fluorescent nor heterogeneous
• the solute must not undergo to photochemical transformations
• the solute must not undertake variable associations with the solvent.
Types of Electronic Transitions
 Chromophore
It is defind as any isolated covalently bonded group that
shows a characteristic absorption in the UV-Vis region.

The functional groups of organic compounds (ketones,

amines, nitrogen derivatives, etc.), responsible for absorption
in UV/Vis are called chromophores. A species formed from a
carbon skeleton transparent in the near UV on which are
attached one or several chromophores constitutes a
chromogene.
 Auxochrome
An auxochrome can be defined as any group which does not
itself act as a chromophore but whose presence brings about
a shift of the absorption band towards the longer wavelength
of the spectrum. –OH,-OR,-NH2,-NHR, -SH etc. are the
examples of auxochromic groups.
There are four types of shifts observed in the UV spectroscopy.
a) Bathochromic effect- This type of shift is also known as red shift.
Bathochromic shift is an effect by virtue of which the absorption
maximum is shifted towards the longer wavelength due to the
presence of an auxochrome or change in solvents. The n to
π* transition of carbonyl compounds observes bathochromic or red
shift.
b) Hypsochromic shift- This effect is also known as blue shift.
Hypsochromic shift is an effect by virtue of which absorption
maximum is shifted towards the shorter wavelength. Generally it is
caused due to the removal of conjugation or by changing the polarity
of the solvents.
c) Hyperchromic effect- Hyperchromic shift is an effect by virtue of
which absorption maximum increases. The introduction of an
auxochrome in the compound generally results in the hyperchromic
effect.
d) Hypochromic effect- Hyperchromic effect is defined as the effect
by virtue of intensity of absorption maximum decreases.
Hyperchromic effect occurs due to the distortion of the geometry of
the molecule with an introduction of new group.
 Factors Affecting lmax
There are a range of factors that can affect the ultraviolet and visible
absorption characteristics of an organic compound.

• Solvent
• Concentration of the sample
• pH of the sample
• Temperature of the sample.

Uncontrolled changes in these factors can introduce inaccuracy and

imprecision by altering the position (lmax) and intensity (lmax) of the
absorption peaks of the chromophore in the compound. The analyst
must control these parameters if comparison of spectra is to be
meaningful and if quantitative results are to be valid.
With the job of the analyst in mind, sample and standard preparation
will play a key role particularly in regard to using standardized
conditions.
Effect of Solvent
The choice of solvent can shift peaks to shorter or longer wavelengths.
This will depend on the nature of the interaction of the particular solvent
with the environment of the chromophore in the excited state of the
molecule.
Depending on the chromophore in the particular analyte, changes in the
polarity of the solvent can influence shifts to longer or shorter
wavelengths. For instance, it is usually seen that ethanol solutions give
longer wavelength maxima than hexane solutions.

Effect of Concentration
As you might expect, sample concentration is proportional to the
intensity of the absorption. At high concentrations however, molecular
interactions (for example, polymerization) can take place causing
changes to the position and shape of absorption bands. Such an
outcome can affect the linearity of the relationship between sample
concentration and absorbance (remember Beer’s Law).
Such effects need to be identified and taken into consideration for
quantitative work.
Effect of Sample pH
The pH of the sample solution can have a significant impact on absorption spectra.
The mechanism for this is primarily a shift in the equilibrium between the different
chemical forms of an analyte. To illustrate this, pH indicators used in acid/base
titrations change colour at a particular pH because the chemical form of the
indicator-compound undergoes a change at this point.
If pH is known to be a factor, a remedy is to prepare the sample in a suitable buffer
solution so as to maintain the pH at a steady value. The buffer though needs to be
transparent over the wavelength range of the measurements – if the buffer absorbs
radiation, absorbance readings attributed to the analyte may be higher than they
should because the buffer and analyte absorptions will add together at each
wavelength.

Effect of Sample Temperature

Temperature impacts absorption measurements by various means:
• Expansion or contraction of the solvent - leading to lower/higher concentrations
and absorbances, a particular issue with some organic solvents.
• Shifts in equilibria between the chemical forms of an analyte – the nature of the
absorbing species may be changed.
• Changes to reaction rates - enzymatic reactions are particularly sensitive to
temperature.
• Variation in refractive index within the sample solution due to convection currents
– refractive index affects absorbance particularly for some organic solvents.
Woodward-Fieser rules for prediction of λmax
The base value for:
• Butadiene system or acyclic conjugated
diene = 217 nm.
• Homoannular conjugated diene (conjugated
double bond in same ring) = 253 nm.
• Heteroannular conjugated diene
(conjugated double bond in different rings)
= 215 nm.

Increment for each substituent:

• Double bonds extending conjugation = + 30
nm

• Exocyclic double bond = + 5 nm

• Each alkyl substituent or ring residue = + 5

nm
Substituent/Auxochrome effect:
Introduction, types of vibration, selection rules, Hooke’s law, problem,
identification of functional groups like C-C, C=C, -OH, -NH2 and -C=O.
 Introduction
• It is the interaction of IR radiation with matter to determine its
structure and properties.
• It causes vibration of bonds in a molecule and hence the name
vibrational spectroscopy.
• This technique can determine the functional groups present in the
structure of a compound.
• Infrared spectra is due to change in vibrational energy accompanied
by changes in rotational energy.
• It is also called vibrational-rotational spectroscopy.
The IR region is divided into three main regions
• Near IR (overttone region) 0.8 - 2.5µ (12500 - 4000 cm-1)
• Middle IR (vibration-rotation region) 2.5 - 15µ (4000 - 667 cm-1)
• Far IR (rotation region) 15 - 200µ (667 - 50 cm-1)

The main region of interest for analytical purpose :

4000 - 667 cm-1
 Basic Principles
Ø Atom or groups in a molecule are connected by bonds.
Ø These bonds are not rigid in nature. Because of the continuous
motions of the molecule, they maintain some vibrations with
frequency called NATURAL FREQUENCY.
Ø When IR radiation is passed the absorption of infrared radiations
causes an excitation of the molecule.
Ø Then a peak is observed as
APPLIED FREQUENCY=NATURAL FREQUENCY
Ø Each vibrational level is associated with a number of closely
spaced rotational levels. So IR spectra is considered as
VIBRATIONAL ROTATONAL spectra.
Ø Every bond or portion of a molecule or functional group requires
different frequency for absorption.
ØHence characteristic peak is observed for every functional group of
a molecule. It is FINGERPRINT of a molecule.
 Basic Principles
Ø Generally two regions are seen in IR spectra.
• GROUP FREQUENCY REGION
• FINGER PRINT REGION
Ø All the bonds in a molecule are not capable of absorbing infrared
energy. Only those bonds which are accompanied by a change in
dipole moment will absorb in the IR region.
Ø Such vibrational transitions are called as infrared active
transitions.
Ø The vibrational transitions which are not accompanied by a
change in the dipole moment are called as infrared inactive
transitions.
Example:
Vibrational transitions of C=O, N-H, O-H shows change in dipole
moment. Vibrational transitions of C=C bonds in symmetrical
alkenes and alkynes does not show any change in the dipole
moment.
 Molecular Vibrations

Ø When radiations with frequency range less than 100 cm-1 are
absorbed, molecular rotations takes place and discrete lines are
formed in the spectrum.
Ø When more energetic radiation in the region 104 to 102 cm-1 are
passed through the sample molecular vibrations are set in.
Ø Vibrational spectra appears as vibrational–rotational bands, since
a single vibrational energy is accompanied by a large number of
rotational energy changes.
The vibrations for the molecules may be of two types. They are
1. Stretching Vibrations : In these vibrations distance between
the two atoms increases or decreases. But the atoms are in the
same axis. Bond length is altered.

2. Bending Vibrations : The positions of the atoms changes

relative to the original bond.

In Plane Bending Out of Plane Bending

 symmetric asymmetric In-plane scissoring

Out-of-plane twisting In-plane rocking Out-of-plane wagging

 Simplified Model for Vibrational Interactions

For modelling the vibrations of

bonds, a harmonic oscillator
should be imagined, which
comprises two masses linked
by a spring and able to slide in
a plane without friction. If the
two masses are displaced by a
value x0 with respect to the
equilibrium distance Re and
then the system is released, it
will begin to oscillate with a
period that is a function of the
constant relating to the stiffness
of the spring, named force
constant k (N/m) and of the size
of the masses involved.
The frequency, which is independent of the elongation, is given by Hooke’s
law in which µ (kg) represents, as indicated, the reduced mass of
the system.

k: force constant, µ: reduced mass of a diatomic molecule

The vibrational energy of this simple model can vary in a continuous

fashion. Following a short elongation x0 with respect to the distance
from equilibrium Re, Evib becomes:
 Vibrational Frequency

Ø The value of stretching vibrational frequency of a bond can

be calculated by using HOOKE’S LAW.

Ø Value of vibrational frequency or wave number depends

upon:
• Bond strength .
• Reduced mass
• Hydrogen bonding

Examples:
• C=C has higher vibrational frequency than C-C stretching.
• O-H has higher vibrational frequency than C-C bonding.
• F-H has higher vibrational frequency than O-H stretching.
Problems

1. Calculate the force constant of the bond between H and Br using

absorption maximum = 3.77 µm, velocity of light = 3 ´ 108 m/s, masses
of H and Br atom = 1.008 and 79,9 amu respectively.

2. The fundamental vibrational frequency of HCl is 8.667 ´ 1013 s-1.

calculate the force constant. Mass of H and Cl are 1.008 g/mol and 35.5
g/mol respectively.

3. The fundamental frequency of CO molecule is 2170.2 cm-1. Calculate

the force constant of the given molecule. Velocity of light is 3 ´ 108 m/s.

4. Calculate the absorption frequency for the C-H stretch with a force
constant of k = 5.0x102 N/m.
 Dipole moment and infrared spectrum
All types of molecules cannot interact with IR radiation. Only those molecules
which undergo a net change in dipole moment because of its vibrational or
rotational motion absorb in the IR region.
Homonuclear diatomic molecules like H2 , O2 , N2 or Cl2 do not show change in
dipole moment during vibration and hence they do not absorb in the infrared
region. Such molecules are termed as infrared inactive.

Infrared radiation is absorbed when the oscillating dipole moment interacts with the
oscillating electric vector of the infrared beam. For such an interaction to occur, the
dipole moment of the molecule in the ground state and the vibrational excited state
should be different. The greater the difference in these dipole moments, the more
intense is the absorption.
For example, the symmetrical stretching vibration of the C–C bond in ethylene can
be represented as
Frequency Range in IR
 NMR Spectroscopy

Nuclear magnetic resonance (NMR) involves the interaction between

an oscillating magnetic field of electro-magnetic radiation and the
magnetic energy of the hydrogen nucleus or some other type of nuclei
when these are placed in an external static magnetic field. The sample
absorbs electro-magnetic radiation in radio-wave region at different
frequencies since absorption depends upon the type of protons or
certain nuclei contained in the sample.
Introduction
• When a charged particle such as a proton spins on its axis, it
creates a magnetic field. Thus, the nucleus can be considered to
be a tiny bar magnet.
• Normally, these tiny bar magnets are randomly oriented in space.
However, in the presence of a magnetic field B0, they are oriented
with or against this applied field.
• The energy difference between these two states is very small
(<0.1 cal).
https://www.youtube.com/watch?v=_3OSb37FGe8
 Nuclear Spins in B0
 For 1H and 13C, only two orientations are allowed.

 In an applied field strength of 7.05T, which is readily available with present-

day superconducting electromagnets, the difference in energy between
nuclear spin states for
• 1H is approximately 0.0286 cal/mol, which corresponds to
electromagnetic radiation of 300 MHz (300,000,000 Hz)(300MHz)
• 13C is approximately 0.00715 cal/mol, which corresponds to
electromagnetic radiation of 75MHz (75,000,000 Hz)(75 MHz)
 Chemical Shift
• If the resonance frequencies of all protons in a molecule were the
same, they would all absorb at the same field strength and the
spectrum would consist of a single signal. However, it is not so.
The precessional frequency of all the protons in the same external
magnetic field is not the same.

• The frequency at which a proton absorbs depends upon the

magnetic field that the proton feels or experiences. This in turn
depends upon the environment of the proton, electron density
around the proton, presence of nearby protons and various other
factors.

• When a molecule is placed in a magnetic field, the electrons

rotating around it generate their own magnetic field. This induced
magnetic field may align with the applied field or may oppose it.
• If it aligns with the applied field, the proton feels a higher field
strength and is said to be deshielded.

• If the induced magnetic field opposes the applied field, the proton
experiences a lesser magnetic field and is said to be shielded.

• Shielding shifts the absorption up field and deshielding shifts the

absorption down field to get an effective field strength necessary
for absorption.

• Such shifts in the position of NMR spectrum (as compared to a

standard reference) which arise due to the shielding and
deshielding of proton by the electrons are called chemical shifts.
Measurement of chemical shift
• The standard used universally to measure chemical shift is
tetramethylsilane (TMS).

• TMS has 12 equivalent protons and hence a sharp single signal

is obtained. Owing to the low electronegativity of silicon, the
shielding of equivalent protons in tetramethylsilane is greater than
most organic compounds. The NMR signal of TMS is taken as
reference and the signals of other compounds are measured
relative to it.

• This difference in the absorption position of the proton with

respect to TMS signal is called chemical shift.
NMR spectrometer