SPECTROSCOPY

Peter Chimtali 1
It is a branch of science that deals with interaction of matter
with light or electromagnetic radiation.

Spectrometry is the spectroscopic technique used to assess the

concentration or amount of a given species. In those cases, the
instrument that performs such measurements is a spectrometer or
spectrograph

Spectrum – graph or plot of intensity of absorbed/

emitted radiation by sample verses frequency or
Wavelength
Chemists use spectroscopy (IR, UV-Vis, NMR) to determine
molecular structure. The technique measures the amount of
radiation a substance absorbs at various wavelengths (based
on the quantization of molecular energy levels). Molecules
exist in quantized energy states (rotational, vibrational, and
electronic quantum states). Molecules absorb discrete
amounts of energy (ΔE) and are excited to higher energy
states. We can use electromagnetic radiation to excite
molecules.
 Classification of Methods
The type of spectroscopy depends on the physical quantity
measured. Normally, the quantity that is measured is an
intensity, either of energy absorbed or produced.
Most spectroscopic methods are differentiated as either
atomic or molecular based on whether or not they apply to
atoms or molecules. Along with that distinction, they can be
classified on the nature of their interaction:
Spectroscopic techniques can be divided into two:
 Atomic spectroscopy (atomic absorption and atomic emission)
Absorption spectroscopy
It uses the range of the electromagnetic spectra in which a
substance absorbs. This includes atomic absorption
spectroscopy and various molecular techniques, such as infra-
red spectroscopy in that region and Nuclear Magnetic
resonance spectroscopy in the radio region
Emission spectroscopy
It uses the range of electromagnetic spectra in which a
substance radiates (emits). The substance first must absorb
energy. This energy can be from a variety of sources, which
determines the name of the subsequent emission, like
luminescence. Molecular luminescence techniques include
spectroflourimetry.
 Molecular spectroscopy (UV-Vis, IR, NMR, MS)
Electromagnetic Radiation
Electromagnetic radiation is a form of energy that is
transmitted through space at enormous velocities.
Electromagnetic radiation, or light, is described by the
properties of both waves and particles nature.
Wave nature of light
In dealing with phenomena such as reflection, refraction,
interference and diffraction, electromagnetic radiation is
conveniently modelled as waves.
An electromagnetic radiation is characterised by several
fundamental properties: frequency, velocity, amplitude,
phase angle, polarization and direction of propagation.
Properties of Waves

 Wavelength (λ) – Distance between two nearest

crest or troughs
 Frequency – Number of wave cycle in a given time,
measured in Hertz (Hz)
 Amplitude – Wave’s height or length

 Wavenumber: is the reciprocal of wavelength

The speed of a wave is the distance one wave travels in a
given time period. The relation between wavelength,
frequency, and speed is

Speed = wavelength × frequency

 Particle Nature of Light
Each light particle, or photon, carries a discrete amount of
energy. This energy is proportional to the frequency of the light,
and is therefore inversely proportional to the wavelength of light.

hc
E photon  hv 


h = Planck’s Constant, 6.626 x 10-34 Js

Photon: A particle-like unit of electromagnetic radiation

(Light), emitted or absorbed by an atom when an
electrically charged electron changes states
 Example:

The energy difference between the 3p and the 3s orbitals in

a sodium atom is 2.107 eV. Calculate the wavelength (in nm) that
would be absorbed in exciting the 3s electron to the 3p state
(1 eV = 1.60 X 10–19 J)

Answer:   590nm
 Electromagnetic Radiation and its interaction with matter
The interactions of radiation and matter are the subject of
spectroscopic studies. The most interesting types of interactions in
spectroscopy are absorption and emission of radiation molecular
or atomic species of interest which involve transitions between
different energy levels of the chemical species.
 Electromagnetic spectrum:

 The arrangement of all types of electromagnetic radiation in

order of their increasing wavelength or decreasing frequency
is known as complete electromagnetic spectrum.

 The visible spectrum represents only a small portion of the

electromagnetic spectrum.

Infra-red radiation have longer wavelength and are thus, less

energetic. Cosmic rays carry high energy while radiowaves are
least energetic. Microwaves have larger wavelength and are
used in telephone transmission.
 Energy Levels in the atom
As we considered, the electrons in the atom can only occupy quantized
orbits, i.e. energy levels. All electrons prefer to be in the lowest
(unoccupied) energy level (lowest potential energy). That is why an
electron at a higher energy level eventually falls in a lower level
releasing a photon. We say there is a potential well in the atom
where the top of the well is at zero potential and the other energy
levels are at negative potential. The ground level will be at the
lowest potential energy

E = 0 eV
E = -1.51 eV
E = -3.40 eV Higher Energy Levels

E = -13.61 eV Ground State

 Ionization Energy
So, the energy levels are a bit like steps of a ladder that the electrons can
climb, if provided with the right energy. What would happen if an
electron is given enough energy to climb to the last energy level
(E = 0)?
• The electron will escape from the atom, as it has escaped from the
potential well that holds it in the atom
• The atom is left with one less electron in its electron cloud, so it
becomes a positive ion
• The energy needed for this jump is called IONIZATION ENERGY

Ionization Energy is the energy required to cause an electron to

escape from the atom
 Excitation Energy
But what can give this energy to the electron to move up the ladder?
• An electric charge (discharge tubes)
• Heating the metal (thermionic emission)
• A photon with just the right energy can hit an electron giving its
energy for the electron to jump to the next energy level

The energy of the photon

must be exactly the same
as the energy gap
between the two level
 Absorption
Here, electromagnetic energy is transferred to the atoms or
molecules constituting the sample; as a result, these
particles are promoted from a lower energy state to higher-
energy states, or excited states. Note that at room
temperature, most substances are in their lowest energy or
ground state
energy
4f n=1
4d DE
n=4 n=2
4p
3d n = 3,
4s etc.
Energy

n=3 3p
3s
n=2 2p
2s
n=1 1s
 Emission

when an atom or molecule in an excited state returns to a

lower energy state, the excess energy often is released as a
photon, a process we call emission.
Emission spectroscopy is a spectroscopic technique which
examines the wavelengths of photons emitted by atoms or
molecules during their transition from an excited state to a
lower energy state.
Each element emits a characteristic set of discrete wavelengths
according to its electronic structure, and by observing these
wavelengths the elemental composition of the sample can be
determined. Emission spectroscopy developed in the late 19th
century and efforts in theoretical explanation of atomic emission
spectra eventually led to quantum mechanics.
 Energy Levels in Atoms
We can think of an atom as consisting of a positive nucleus (protons and
neutrons) surrounded by negative electrons. The electrons can be thought of as
“orbiting” the nucleus, but are only allowed in certain orbits (or energy levels).
A photon with exactly the right energy can excite the electron from one level to
another. The electron will drop back to the “ground state”, and emit photons
with specific energies as it does so.
 Energy transitions and photons
The energy of photon that can interact with a level jump just depends on
the energy difference between the levels. Levels can be skipped.
Emission or absorption of radiation occurs only when
an electron makes a transition from one orbit to
another.
The frequency f of the emitted (absorbed) radiation is
given by
DE  hf  Ef  Ei
where DE : changeof energy
h : Planck's constant
Ef : finalenergy state
Ei : initialenergy state
Note:

 If Ef > Ei Absorption of EM radiation

 If Ef < Ei Emission of EM radiation
  Figure below shows diagrammatically the various energy levels in the
hydrogen atom.
n En (eV)
Figure 4  0.0 Free electron

0.544 excited state

0.85 3 excited state
th
5
4 rd

3 1.51 2nd excited state excited state

Ionization energy
is defined as the
energy required by
an electron in the
2 3.40 1st excited state
is defined as the
energy levels
ground state to that higher
escape completely than the
Excitation energy ground state.
from the attraction is defined as the energy
of the nucleus. required by an electron that
raises it to an excited state
from its ground state. is defined as the
lowest stable
An atom energy state of an
becomes ion. 1 13.6 Ground state atom.
Example 1 :
The electron in the hydrogen atom makes a transition from the
energy state of 0.54 eV to the energy state of 3.40 eV.
Calculate the wavelength of the emitted photon. (Given the
speed of light in the vacuum, c =3.00108 m s1 and Planck‟s
constant, h =6.631034 J s)
Solution :
The change of the energy state in joule is given by

Ei  0.54 eV; Ef  3.40eV

DE  Ef  Ei DE   3.40 0.54

 2.86 1.601019 
DE  4.58 1019 J
Therefore the wavelength of the emitted photon is

DE 
hc
4.5810 19

6.6310 3.0010 
34 8

 
  4.34 10 m 7
Line spectrum
 The emission lines correspond to the photons of
discrete energies that are emitted when excited atomic
states in the gas make transitions back to lower energy
levels.
 Figure below shows line spectra produced by emission in
the visible range for hydrogen (H), mercury (Hg) and
neon (Ne).
Hydrogen emission line spectrum
 Emission processes in hydrogen give rise to series, which are
sequences of lines corresponding to atomic transitions.
 The series in the hydrogen emission line spectrum are
 Lyman series involves electron transitions that end at the
ground state of hydrogen atom. It is in the ultraviolet (UV)
range.
 Balmer series involves electron transitions that end at the
1st excited state of hydrogen atom. It is in the visible light
range.
 Paschen series involves electron transitions that end at
the 2nd excited state of hydrogen atom. It is in the infrared
(IR) range.
 Brackett series involves electron transitions that end at the

3rd excited state of hydrogen atom. It is in the IR range.

 Pfund series involves electron transitions that end at the
4th excited state of hydrogen atom. It is in the IR range.
 Figure belwo shows diagrammatically the series of hydrogen
emission line spectrum.
n E n (eV)
 0.0 Free electron

5 Pfund series0.544rd excited state


th
4 0.85 3 excited state
Brackett series 1.51 2 nd excited state
3
Paschen series

2
Balmer series
3.39 1 st excited state

Lyman series

1 13.6 Ground state

MOLECULAR ENERGY LEVELS
Radiation can be absorbed or emitted if the molecule changes
any of its energy states.
 The relative energy of the spacings between energy levels for
MOLECULAR
 various ENERGY LEVELS _ CONT’D
types of ansitions in a molecule are in the order:

<< <<
Rotational Vibrational Electronic
Transition Transition Transition

1-20 cm-1 2000-4000 cm-1 10000-50000 cm-1

The various types of energy transitions occur in different regions of the EM-
spectrum and do not overlap
 MOLECULAR ENERGY LEVELS _ CONT’D

Vibrational
Transition

2000-4000 cm-1

Infrared

Rotational Electronic
Transition Transition

1-20 cm-1 10000-50000 cm-1

Microwave UV-Visible
• Electronic,
vibrational and
rotational energy
levels are super
imposed
• The absorption
spectrum of a
molecule is
determined by all
allowed
transitions
between pairs of
energy levels V=Vibration quantum number
that interact with
the radiation field J=Rotation quantum number
 UV/VIS Spectroscopy:

• Absorbed by electron transitions. It basically involves the

spectroscopy of photons and spectrophotometery. It uses light in
the visible and adjacent near ultraviolet (UV) and near infrared
(NIR) ranges.
• Sunburn is primarily an effect of UV radiation, and ionisation
produces the risk of skin cancer.
• Absorption of visible light causes heating , but not ionisation
• UV/Vis spectroscopy is routinely used in the quantitative
determination of solutions of transition metal ions and
highly conjugated organic compounds.

• For the quantitative measurements, Beer-Lambert law is

followed. The Beer-Lambert Law is useful for
characterizing many compounds but does not hold as a
universal relationship for the concentration and absorption
of all substances.
 Infra-red Spectroscopy:

The energy level of infrared corresponds to the energy

required to cause molecular vibrations. Vibrations arise as
molecular bonds are not rigid but behave like springs
(IR spectroscopy) is the subset of spectroscopy that
deals with the infrared region of the electromagnetic spectrum.
It covers a range of techniques, the most common being a
form of absorption spectroscopy.
Vibrational transitions are Sub-divided into two classes:

Stretching : symmetric and asymmetric

Bending: scissoring, rocking, wagging and twisting

Stretching frequencies are higher than corresponding bending
frequencies - it easier to bend a bond than to stretch or
compress it. Bonds to hydrogen have higher stretching
frequencies than those to heavier toms. Triple bonds have
higher stretching frequencies than corresponding double bonds
which in turn have higher frequencies than single bonds
 As with all spectroscopic techniques, it can be used to
identify compounds or investigate sample composition.

Infrared spectroscopy offers the possibility to measure

different types of inter atomic bond vibrations at different
frequencies. Especially in organic chemistry the analysis of IR
absorption spectra shows what type of bonds are present in
the sample.

Infrared spectroscopy exploits the fact that molecules

have specific frequencies at which they rotate or vibrate
corresponding to discrete energy levels.
 MICROWAVE SPECTROSCOPY
Quantum energy of microwave photons ( 10-5-10-3eV)
matches the ranges of energies separating quantum
states of molecular rotations.
Energy levels of a molecule free to rotate are quantized and
transition between these levels give rise to the rotational
spectrum of a molecule. The electromagnetic radiation
emitted for these transition lies in the microwave region with
wavelengths of the order of 1 cm; hence, rotational
spectroscopy is also known as microwave spectroscopy.
 Transmittance and Absorbance:
It is difficult, if not impossible, to measure actual absorbance of
light. Instead, we measure transmittance or the fraction of light
that is able to pass through a solution of molecules. A
spectrophotometer measures the intensity of light entering a
sample and compares this to the intensity of light emerging
from the sample
 Io I

Io= intensity of light entering the sample

I= intensity of light leaving the sample

I
T
I
 Factors that affect Transmittance
• Concentration:
Each molecule can absorb light. As you increase the
number of molecules in the solution, you also increase the
photons absorbed. Therefore, as you increase the
concentration of a sample, you decrease the transmittance.

• Length of the sample path: By increasing the pathway

that the light must travel through your sample, you are
increasing the number of molecules that will interact with
the light; in effect, you are increasing the apparent
concentration.
Specific properties of the molecules. Molecules absorb
light at different efficiencies and at different energies.
Therefore, the transmittance will be dependent upon the
specific molecule in solution and the wavelength of light
being passed through the sample.
 BEER-LAMBERT LAW

Interaction of light with matter

Definition of absorbance & transmittance

Io I

I
% T  100 %
A   log T I
I
A   log
I
Absorbance (A) and transmittance (T) are unitless
Example:
The intensity of the incident light on a sample is 0.5 wm-2 and
the intensity of light entering the detector is 0.36 wm-2
a) Calculate the transmittance (T=0.72)
b) Determine the absorbance (A=0.14)
 Beer-Lambert’s law
Absorbance Vs. Concentration (c ) Absorbance Vs. Path length (b)

Io I [1] Io I [1]
s
s

Io I [2] Io I [2]
s s

Ac Ab
This is not true at high Path length- the distance that light
travels in the sample
concentrations

Unit of concentration = M Unit of path length = cm

 Beer-Lambert’s law
Ac Ab
A  bc

A abc

a – Molar absorptivity
Units of molar absorptivity:
unit of A  (unit of a)(unit of b)(unit of c)
unit of A
unit of a 
(unit of b)(unit of c)
 unit of A
unit of a 
(unit of b)(unit of c)

unitless 1
unit of a    cm1M 1  M 1cm1
(cm)( M ) (cm)( M )

“A” is unitless but “a” has units M-1cm-1

 Definition of molar absorptivity

A abc
A
a
( b)(c)

a - is a measure of the amount of light absorbed per unit

concentration and path length
A compound with a high molar absorptivity is
very effective at absorbing light (of the appropriate
wavelength)
 Sample Problem

Find the molar absorptivity of the a 0.5 M solution

whose absorbance is 0.77,When measured in a
tube of path length 2 cm?

Given:
Concentration of the solution = 0.5 M =c
Absorbance = 0.77= A
Path length = 2 cm = b

To be found:
Molar absorptivity of the solution= ?= a
 Sample Problem
Methodology:
Beer’s law
A abc

Calculation:
A
a
( b)(c)
0.77
a  0.77cm1M 1  0.77M 1cm1
( 2 cm)(0.5M )
Determination of Concentration Using a Standard Curve:
Standard curves are generated when a is not known and/or to
minimize experimental error. Determine absorbance of known
concentrations. Then plot a graph of absorbance against
concentration. This gives a straight line. The line should have a
y intercept of zero (when the concentration is zero, there
should be zero absorbance) and the slope of the line is equal
to a.
Y=mx + c: y is the absorbance, m is a, x is
concentration, and c is the y-intercept
 0.7
0.6
Absorbance

0.5
0.4
0.3
0.2
0.1
0
0 0.1 0.2 0.3 0.4 0.5 0.6
Concentration, M
DEVIATION FROM BEER’S-LAMBERT LAW
In beer’s law it states that if we plot absorbance A against
concentration C a straight line passing through origin is obtained,
but usually a deviation from linear relationship between
concentration and absorption and an apparent failure of beer’s
law.

There are two types of deviation:

i) POSITIVE DEVIATION:
when a small change in concentration produces a greater
change in absorbance

ii) NEGATIVE CHANGE

when a large change in concentration produces a smaller
change in absorbance.
Sources of Error in Beer’s Law Measurements:
Beer’s Law is only true for dilute solutions--the exact range of
solutions must be determined experimentally. Beyond this range,
measurements and calculations using Beer’s Law will be
erroneous.

Other common sources of error include the use of dirty cuvettes

(brings impurities), poorly mixed solutions, poor pipetting
techniques, and incorrect light source or wavelength (requires the
use of monochromatic light).