Principle of Spectroscopy

Syllabus:
Introduction: Principle of spectroscopy, Definition, Origin of spectrum,
Classification of spectroscopy – atomic and molecular, selection rules. Table
of relation between electromagnetic spectrum, types of spectroscopy and
energy changes. Principle and application of Flame photometry and
Fluorescence spectroscopy.
 Introduction
• Spectroscopy deals with the study of interaction of
electromagnetic radiation with matter. Electromagnetic radiation is
a simple harmonic wave of electric and magnetic fields fluctuating
orthogonal to each other.
• Merits of spectroscopic techniques over the classical techniques-
1. It takes less time.
2. Very small amount of compounds are required for studies.
3. It is fast and economical and reliable.
4. It is nondestructive ie compounds being studies remain unchanged
during studies and can be reused.
5. In this method information is recorded in the form of permanent
chart.
 Electromagnetic Radiation:
• Energy of an electromagnetic radiation is given by 𝐸=ℎν=ℎ𝑐𝜆, where h is
Planck’s constant
 Characteristics of electromagnetic radiation

 These radiations are produced by oscillation of electric charge and

magnetic field residing on this atom. The electric and magnetic
components are mutually perpendicular to each other and are
coplanar.
 These are characterized by their wavelength or frequencies or wave
number.
 The energy carried by an electromagnetic radiation is directly
proportional to its frequency. The emission or absorption of
radiation is quantized and each quantum of radiation is called a
photon.
 All types of radiation travel with the same velocity and no medium
is required for their propagation. They can travel through vacuum.
 When a visible light (a group of electromagnetic radiation) is passed
through a prism,it is split into seven colors which correspond to
definite wavelengths. This phenomenon is called as dispersion.
The energy changes in a molecule are specified using these terms:
 Amplitude: It is the distance from the maximum vertical displacement of the wave
to the middle of the wave. This measures the magnitude of oscillation of a particular
wave. Large amplitude means higher energy and lower amplitude means lower
energy. Amplitude tells us the intensity of brightness of a waves (light) in comparison
with other waves.
 Wavelength: It is the distance of one full cycle of the oscillation. It is denoted by
lambda ‘λ‘. Longer wavelength waves such as radio waves carry low energy and
shorter wavelength waves such as X-rays carry higher energy which can be harmful
for our health. The relationship between the speed of light ‘C’, the frequency ‘ʋ’ and
wavelength ‘λ‘ is given as ….C = λ ʋ
 Frequency: It is the number of cycle per second. It is denoted by nu ‘ʋ’. It is
expressed as sec -1 or Hertz (Hz). It is directly proportional to energy ‘E’ as,
E=hʋ …………(ii)
Where ‘h’ is Plank’s constant (6.63 x 10 -34 J)
 Period: It is the time a wave takes to travel one wavelength. It is denoted by ‘T’ and
is measured in seconds (s).
 Velocity: The velocity of a wave is denoted by ‘v’ and is expressed as
V =λʋ …………(iii)
For electromagnetic wave, the velocity in vacuum is 2.99 x10 8 m/s.
 Wavenumber: It is the number of waves per centimeter in vacuum. It is denoted by
nu bar ___ it is the reciprocal of wavelength and is measured in cm -1.
 Origin of electromagnetic radiations

As per Max Planck while studying the problem of Blackbody radiation in early
1900s, atoms and molecules can absorb or emit the energy in discrete packets,
called quanta (singular: quantum).

The quantum for electromagnetic energy is called a photon

A molecule can possess energies in different forms such as vibrational energy,

rotational energy, electronic energy, etc.

Electrons resides in different orbits/orbitals surrounding the nucleus,

 Rotational and vibrational energy levels of molecules are also discrete.

A molecule can jump from one energy level to another by absorbing or emitting
a photon of energy that separate the two energy levels as shown in following
diagram.
 Electromagnetic spectrum and the atomic/molecular processes:
Molecules undergo processes like rotation, vibration, electronic transitions, and nuclear
transitions. The energies underlying these processes correspond to different regions in the
electromagnetic spectrum
• Radiofrequency waves: Radiofrequency region has very low energies that correspond
to the energy differences in the nuclear and electron spin states. These frequencies,
therefore, find applications in nuclear magnetic resonance and electron paramagnetic
resonance spectroscopy.
• Microwaves: Microwaves have energies between those of radiofrequency waves and
infrared waves and find applications in rotational spectroscopy and electron
paramagnetic resonance spectroscopy.
• Infrared radiation: The energies associated with molecular vibrations fall in the infrared
region of electromagnetic spectrum. Infrared spectroscopy is therefore also known as
vibrational spectroscopy and is a very useful technique for functional group
identification in organic compounds.
• UV/Visible region: UV and visible regions are involved in the electronic transitions in the
molecules. The spectroscopic methods using UV or visible light therefore come under
‘Electronic spectroscopy’.
• X-ray radiation: X-rays are high energy electromagnetic radiation and causes transitions
in the internal electrons of the molecules.
 Classification of spectroscopy

The various implementations and techniques can be classified in several ways as

under
I) Type of radiative energy
In many applications, the spectrum is determined by measuring changes in
the intensity or frequency of radiative energy such as:
1. Electromagnetic radiation was the first source of energy used for spectroscopic
studies. Techniques that employ electromagnetic radiation are typically
classified by the wavelength region of the spectrum and include microwave,
terahertz, infrared, near-infrared, ultraviolet-visible, x-ray, and gamma
spectroscopy.
2. Particles, because of their de Broglie waves, can also be a source of radiative
energy. Both electron and neutron spectroscopy are commonly used. For a
particle, its kinetic energy determines its wavelength.
3. Acoustic spectroscopy involves radiated pressure waves.
4. Dynamic mechanical analysis can be employed to impart radiating energy,
similar to acoustic waves, to solid materials.
 Classification of spectroscopy
II) Nature of the interaction
(Nature of the interaction between the energy and the material)

1. Absorption spectroscopy: Absorption occurs when energy from the radiative

source is absorbed by the material. Absorption is often determined by
measuring the fraction of energy transmitted through the material, with
absorption decreasing the transmitted portion.

2. Emission spectroscopy: Emission indicates that radiative energy is released

by the material. A material's blackbody spectrum is a spontaneous emission
spectrum determined by its temperature. This feature can be measured in the
infrared by instruments such as the atmospheric emitted radiance
interferometer. Emission can also be induced by other sources of energy such
as flames, sparks, electric arcs or electromagnetic radiation in the case of
fluorescence.

3. Elastic scattering and reflection spectroscopy determine how incident

radiation is reflected or scattered by a material. Crystallography employs the
scattering of high energy radiation, such as x-rays and electrons, to examine the
arrangement of atoms in proteins and solid crystals.
 Classification of spectroscopy
4. Impedance spectroscopy: Impedance is the ability of a medium to
impede or slow the transmittance of energy. For optical applications, this
is characterized by the index of refraction.

5. Inelastic scattering phenomena involve an exchange of energy

between the radiation and the matter that shifts the wavelength of the
scattered radiation. These include Raman and Compton scattering.

6. Coherent or resonance spectroscopy are techniques where the

radiative energy couples two quantum states of the material in a
coherent interaction that is sustained by the radiating field. The
coherence can be disrupted by other interactions, such as particle
collisions and energy transfer, and so often require high intensity
radiation to be sustained. Nuclear magnetic resonance (NMR)
spectroscopy is a widely used resonance method, and ultrafast laser
spectroscopy is also possible in the infrared and visible spectral regions.

7. Nuclear spectroscopy are methods that use the properties of specific

nuclei to probe the local structure in matter, mainly condensed matter,
molecules in liquids or frozen liquids and bio-molecules.
 Classification of spectroscopy
III) Types /classification of spectroscopy based on atomic and molecular level of studies
1.Atomic spectroscopy 2.Molecular spectroscopy.

1) Atomic Spectroscopy-In this case , the energy changes take place at atomic levels . Atoms
of different elements have distinct spectra .Atomic spectroscopy can quantify and
identify a sample composition. It includes Atomic Absorption Spectroscopy (AAS), Atomic
Emission Spectroscopy (AES) and Atomic Fluorescence Spectroscopy.

i. AAS: Atomic absorption spectroscopy is based on absorption of light(uv/Visible) by free

metallic ions. It quantifies amount of light absorbed of ground state atoms in the
gaseous state. It is used for detection of metals.

ii. AES: In AES (Flame emission photometry) atoms are excited by heat of flame to emit
heat. It is used to determine quantity of an element in a sample by measuring the
intensity of light emitted. Instead of heat of flame if heat from plasma is used for
emission, then it is plasma emission and if an arc is used, it is arc emission spectroscopy
9photometry). AES is used to detect multiple elements simultaneously.

iii. AFS: It is a beam of light that excites the atoms in the analytes causing them to emit
light. The fluorescence from a sample is then analyzed using a florometer. It is commonly
used to analyze organic compounds.
Classification of spectroscopy
Classification of spectroscopy
Classification of spectroscopy
 Application of Molecular Spectroscopy

1. It is used in structural investigation. Electronic energy levels, bond

lengths, bond angles and strength of bond can be determined with the
help of molecular spectroscopy.
2. It can be used to monitor changing concentrations of reactants and
products in kinetic studies.
3. Emission spectroscopy is used for the study of energetically exited
reaction products.
4. It helps to understand the colour in the world around us.
 Selection Rule:

• A selection rule describes how the probability of transitioning

from one level to another cannot be zero.
• It has two sub-pieces: a gross selection rule and a specific
selection rule.
• A gross selection rule illustrates characteristic requirements for
atoms or molecules to display a spectrum of a given kind, such
as an IR spectroscopy or a microwave spectroscopy. Once the
atom or molecules follow the gross selection rule, the specific
selection rule must be applied to the atom or molecules to
determine whether a certain transition in quantum number
may happen or not.
• Selection rules specify the possible transitions among quantum
levels due to absorption or emission of electromagnetic
radiation.
 Selection Rule:
• Atoms are described by the primary quantum number n, angular
momentum quantum number L, spin quantum number S, and total angular
momentum quantum number J. Based on Russell-Saunders approximation
of electron coupling, the atomic term symbol can be represented as 2S+1LJ.

1. The total spin cannot change, ΔS=0;This rules says spin angular
momentum of an electron does not change during the absorption or
emission of light
2. The change in total orbital angular momentum can be ΔL=0, ±±1, but L=0
↔↔ L=0 transition is not allowed. When ΔL=+1, it means that an
electron jumps from a lower to higher energy state. When ΔL=-1, it means
that an electron jumps from a higher to lower energy state. Thus, this
rules tells us an electron cannot jump between two orbitals that differ by
more than one orbital quantum number and the changes in ‘l’ cannot be
equal to zero.
3. The change in the total angular momentum can be ΔJ=0, ±±1, but J=0
↔↔ J=0 transition is not allowed. The total angular momentum is the
sum of the spin and orbital angular momenta for the electrons in an
 Selection Rule:

4. The change in the magnetic quantum number is either zero , or +1,or ,-1.
Δml=0, ±±1. A 2s orbital has a principal quantum number n=2, the orbital
quantum number l=0 , and magnetic quantum number Δml=0.In contrast , a
2p orbital has px , py and pz orbitals .pz orbital has n=2,l=1 and ml=+1.The py
orbital has n=2,l=1 and ml=0.The px orbital has n=2,l=1 and ml=-1.These 3
orbitals differ only in the magnetic quantum number .If we have an electron
in 2pz orbital ,and if we follow selection rule ,we can see that the electron
can jump from any of three 2p orbital to 2s orbital .This can be explained as
follows, orbital quantum number changes from l=1 to l=0.Thus ΔL=-1.This
means that the transition from 2pz or 2py or 2pxorbitals to 2s orbital is an
allowed transition. On the contrary ,if an electron can transit from 2pz orbital
to 2py orbital or from 2py to 2px orbital ,is this transition allowed or not .The
change in orbital quantum number ΔL=0,in all the above cases .These
transition are known as forbidden transition .The electron cannot jump from
one 2p orbital to other 2p orbital.
Selection Rule:

Allowed Transition
Relation between electronic spectrum, types of spectroscopy and
corresponding energy change
 Applications of Spectroscopy
• Emission Spectroscopy
• Emission Spectroscopy deals with atoms.
• It consists of the excitation of a sample using thermal or electrical energy.
• The emitted radiations are analysed. The method is useful for metal analysis.
• The metal and metalloids can be identified in very low concentration,
approximately 1 ppm.
• The element must be present in solid state.The liquid solutions are evaporated
and then analysed .
• When a sample is excited thermally or electrically , its valence electrons are
excited from lower to higher energy level .The excited electrons rapidly emit a
photon of energy and return to the ground state from the basis of emission
spectroscopy .
• When the flame is used for excitation ,it is termed as “Flame Emission
Spectroscopy” or ‘Flame Photometry’.
 Flame Photometry
• Flame Photometry is based on the measurement of the emitted light intensity , when
a metal is introduced into the flame. The wavelength of the colour gives the
information about the element and colour of the flame gives the information about the
amount of the element present in the sample.
Principle
• In flame photometry the species (metal ions) used in the spectrum are in the form of atoms.
The International Union of Pure and Applied Chemistry (IUPAC) Committee on Spectroscopic
Nomenclature has recommended it as flame atomic emission spectrometry (FAES). The basis
of flame
• Species of alkali metals (Group 1) and alkaline earth metals (Group II) metals are dissociated
due to the thermal energy provided by the flame source. Due to this thermal excitation,
some of the atoms are excited to a higher energy level where they are not stable. The
absorbance of light due to the electrons excitation can be measured by using the direct
absorption techniques. The subsequent loss of energy will result in the movement of excited
atoms to the low energy ground state with emission of some radiations, which can be
visualized in the visible region of the spectrum. The absorbance of light due to the electrons
excitation can be measured by using the direct absorption techniques while the emitting
radiation intensity is measured using the emission techniques. The wavelength of emitted
light is specific for specific elements.
•
Fig 1: A schematic representation of flame photometer

Parts of a flame photometer

1. Source of flame:
2. Nebuliser and mixing chamber:
3. Optical system (optical filter):
4. Photodetector
Fig 2: Brief overview of the process in flame photometer
The flame emission of the alkali and alkaline earth metals in terms of the
emission wavelength and the characteristic colour produced by each element is
shown in table:

Name of the Emitted wavelength Observed colour of

element range (nm) the flame

Potassium (K) 766 Violet

Lithium (Li) 670 Red

Calcium (Ca) 622 Orange

Sodium (Na) 589 Yellow

Barium (Ba) 554 Lime green

Lime green
 Applications of Flame Photometry

• 1. The radiations emitted by the flame photometer are

characteristics of a particular metal. Hence, with the help of flame
photometer presence of any specific element in the given sample
can be detected.
• 2. The concentration of Na+and K+ions are very important in the
human body for conducting various metabolic functions. Their
concentrations can be determined by diluting and aspirating blood
serum sample into flame.
• 3. Soft drink, fruit juices and alcoholic beverages by using flame
photometry to determine the concentrations of various metals and
elements.
• 4. The presence of some of the alkali metals and alkaline earth
metals is critical for soil health .The presence of these metals can
be determined in soil sample by flame photometry. Then, the soil
can be supplied with specific fertilizer.
• Advantage of flame photometry
• 1. The method of analysis is very simple, economical, quick , convenient , selective
and sensitive.
• 2. It is both qualitative and quantity in nature.
• 3. The method can be used to estimate elements which are rarely analyzed. 4.
Even very low concentrations (ppm , ppb) range of metals in the sample ca
be determined and estimated
• 5. The method compensates for any unexpected interfering material present in
the sample solution
• Disadvantage of flame photometry
• 1. The accurate concentration of the metal ion in the solution cannot be
determined and estimated
• 2. It is difficult to obtain the accurate results of ions with higher concentrations 3.
It does not provide the information about the molecular structure of the
compound present in the sample solution
• 4. The element such as carbon, hydrogen and halides cannot be detected by
this method due to their non-radiating nature. A number of metal atoms also
cannot be determined and analyzed.
 Introduction to fluorescence and
phosphorescence
• Fluorescence and phosphorescence are types of
luminescence.
• Luminescence is any emission of light
(electromagnetic waves) from a substance which
does not arise from heating.
• One of the types of luminescence is
photoluminescence (absorption of photon).
• Two types of photoluminescence are (1)
fluorescence (2) phosphorescence.
 Principle Fluorescence Spectrometry
The process in which certain substances when exposed to light radiation of short wavelength
(high frequency) emit light of different frequencies compared to that of incident radiation is
called fluorescence. It stops when the incident radiation is cut –off. The decay period is very
short. When the light is absorbed by a fluorescent substance (fluorophore) ,the excited state of
an atom emits light radiations in several transitions.
• The emitted light has different frequencies for each transition.
• X+hv------X*
• X*--------X’*+hv’
• X’*------X+hv’’
• Where X’*donates the energy states between X and X*.
• The incident and emitted radiation frequencies are different. Fluorescence is a secondary
effect that results from the primary process of absorption of quantum of light by the
substance.
• The emitted radiations are of greater wavelengths than the exciting radiations. After
absorbing the light radiations , the electrons are raised to higher energy levels and then
return to original level in steps. These jumps possess lesser energy than the original jump.
Thus the emitted light has lower frequency than the light absorbed.
• Examples of Fluorescent substances are chlorophyll, calcium fluoride, naphthalene, vapors of
iodine, sodium and mercury.
 Applications of fluorescence Spectrometry
in medicine

• Fluorescence spectroscopy used in drugs and biological samples. It is an

excellent diagnostic and research tool in medical microbiology field with high
sensitivity and specificity.

• It is also used in food analysis, forensic science ,medical diagnosis etc.

• Laser Induced Fluorescence has been used for diagnosing cancer.

• Fluor photometry can be used in determining glucose in blood very accurately.

• The principles of fluorescence can be applied for improved diagnosis and clinical
care in detection and diagnosis of bacterial biofilm from target biological samples.

• Fluorescence spectroscopy is a diagnostic technique with high sensitivity

and specificity for micro-organism associated with infectious diseases such as
viruses, fungi, bacteria and parasites.
Fluorescence Spectrometry