APPLICATIONS OF SPECTROSCOPY

What is emission spectroscopy?

It is the study of Emission spectra which arises when the molecule in excited state returns to the
ground state by emission of light.

What is Flame Emission Spectroscopy?

• When an element is heated to high temperature in a flame it is converted into atoms.

• These atoms absorb heat energy from the flame and get excited.
• The excitation of atoms is due to transition of electrons from lower energy level to higher energy
level.
• The energy required to electronic transition is provided by the high temperature of a flame.
• The life time of the excited electron is very short, hence the excitation is immediately followed
by return of electrons to ground energy level.
• This results in emission of radiation. The intensity of emitted radiation is proportional to the
number of atom excited within the flame and hence is proportional to concentration of element
within the sample solution.
• This study of radiation energy emitted when all the elements is heated in the flame is called
Flame Emission Spectroscopy and the instrument which work on this is called Flame
Photometer.

Discuss the basic principle of Flame Photometry.

Flame Photometry involve measurement of intensity of emitted light radiation by the excited atom
within the flame. Since the intensity of emitted light radiation is directly proportional to the number
of excited atoms, the concentration of elements in the sample can be determined.
Since the wavelength of the emitted light radiation is the characteristics of particular element,
identification of the element is also possible by flame photometry. Thus, flame photometric
technique can be used for quantitative as well as qualitative analysis.
When a liquid sample solution containing metallic salt MA is introduced into a flame the following
events takes place in a rapid succession.

When a liquid sample solution containing metallic salt MA is introduced into a flame the following
events takes place in a rapid succession.

1. The solvent vaporizes leaving behind solid salt.

MA → M (Solid) + A
2. The solid particle forms gaseous molecule which then decompose into gaseous atoms.
M (Solid) → M (Gaseous molecule) → M (Gaseous atom)
3. Some of gaseous atoms absorbs thermal energy from the flame and get excited.
M (Gaseous atom) → M* (Excited gaseous atom)
4. The excited gaseous atom are unstable & short lived they return to the ground state by
emitting UV visible light radiation.
M* (Excited gaseous atom) → M (Gaseous atom) + hv
With the help of a neat labelled diagram, describe various components of flame photometer and
their function.

Flame photometer essential consists of components like:

• Nebuliser (Atomizer)

• Burner

• Filters/ Monochromator

• Detector

• Read out meter

1)Nebuliser: Compressed air is passed into an atomizer. Because of the suction produced by the
compressor this sample solution is sucked into the atomizer. In the atomizer the sample solution is
converted into tiny droplets or aerosol. This process is called nebulisation.
2)Burner: A tiny droplets of the sample is blown by the air into the burner there with mixed with the
fuel supplied with the burner. The mixture is burned in the flame at the top of the burner. There are
two types of burners used Laminar flow (premix burner) and Total consumption burner.
3) Mirror: Light emitted from the flame is reflected by concave mirror placed behind burner. The
reflecting beam is made to fall on the filter or the monochromator.
4)Filter or Monochromator
a) Filter: If a reflected beam consists of small number of wavelengths, then optical filter is used. The
filter allows only the particular wavelength of light radiation (which is characteristic of element
under study) to pass through it but will absorb another wavelength. Since each element requires
specific filter, for analysis of different elements different filters are used.
b) Monochromator: If a reflected beam consists of large number of wavelengths, then
monochromator is used. Monochromator will disperse the reflected light radiation into its individual
wavelengths. By rotating the monochromator, different wavelengths are focused one by one on the
detector. Thus, detection of several elements is possible simultaneously. Examples of
monochromator used are Prism or Diffraction Grating.
5) Detector: commonly used detector is photocell. When a beam of light radiation coming from
monochromator is made to fall on the photo cathode of photocell, electric current is produced. This
electric current is amplified by the electronic amplifier and read on a meter. The magnitude of
electric current produced is directly proportional to the intensity of emitted light radiation which in-
turn is proportional to concentration of element in the sample solution.

Explain Calibration Method and Standard deviation method for finding out the concentration of
unknown sample in flame photometry.

1. Calibration method
A series of standard solution of known concentration is first prepared e.g. 1ppm to 10ppm. These
standard solutions are aspirated one by one into the flame and emission intensity Ei for all the
standard solution is recorded with reference to blank. The calibration graph of emission intensity Ei
against concentration of standard solution in ppm is plotted

The unknown sample solution (x) is aspirated in the flame and its emission intensity (Ei)x is recorded
with reference to blank. Knowing the (Ei)x of unknown sample, concentration of unknown Cx is
obtained from the calibration graph.
2. Standard addition method
when an unknown sample solution is complex in nature, or its exact chemical composition is not
known, then it is difficult to prepare standard solutions. In such case Standard addition method is
used. In this method the emission intensity of unknown sample solution (x) is measured with
reference to blank solution. Series of standard solutions (s) of known and increasing concentration
e.g. 1 ppm to 5 ppm are prepared and to each standard solution fixed amount of unknown sample
solution (x) is added. Then emission intensity of each standard solution are measured with reference
to blank solution.A graph of emission intensity (Ei) is plotted against concentration of standard
solution which gives a straight line graph.
The graph is extrapolated back to intercept to concentration axis the point of intersection on
concentration axis gives the concentration of unknown Cx.

Advantages
1. Since the sample solution isn’t wasted the emission intensity is high and sensitivity of
estimation is also high.

2. It is having low operating cost and maintenance cost.

3. It is reliable and convenient method for the determination of alkali and alkaline earth
metals.

4. It is a controlled method as the temperature of the flame can be adjusted.

Disadvantages
1. It cannot be used to analyse solution with higher concentrations.

2. It cannot be used to detect non-metals such as carbon and halides.

3. It cannot be used for the analysis of those metals which have tendency for incomplete
vaporization.

4. Flame produced is noisy, turbulent and small in cross section.

What are the applications of Flame Photometry?

1. Determination of Ca, Mg in cement.

2. Determination of lead in petrol.

3. Determination of Na & K in fertilizers.

4. It is used in agricultural field for the analysis of water, soil and plant products.

5. It is used in medical field for the testing of urine and blood samples.

6. It is used for the analysis of soft drinks, fruit juices etc.

Fluorescence and Phosphorescence:

Jablonski Diagram
Aleksander Jablonski in 1935 suggested the famous diagram, which makes it possible to explain
spectra of fluorescence and phosphorescence.
Once a molecule has absorbed the energy in the form of electromagnetic radiations it goes to higher
energy level (excited state) from ground state. The electron of the absorbing molecule may jump
from S0 (singlet ground state) to S1 or S2 or S3 (singlet excited state) depending upon the energy of
the light photon absorbed. For each singlet excited state (S1, S2, S3), there is a corresponding triplet
excited state (T1,T2,T3). The activated molecule returns to the ground state by scattering its energy
through the following processes.
I) Non-radiative transitions:
These transitions are from higher excited states (S3, S2 or T3,T2) to the first excited state (S1 or T1)
which doesn’t involve emission of any radiation.
This happens through any of the following processes
a) Internal conversion (IC):The transition between the singlet states viz S3 to S2, S2 to S1 and between
triplets states T3 to T2, T2 to T1are called as internal conversion. It occurs in less than
10-11.
b) Inter-system crossing (ISC):The transition between singlet states and Triplet state viz S2 to T2, S1 to
T1 are called as Inter-system crossing.
c) Vibrational Relaxation is the most common non-radiative transition.
II) Radiative transitions:
In these transitions, the activated molecule returns from the singlet excited state (S1) and triplet
excited state (T1) to the singlet ground state (S0) which involves the emission of radiation. There are
two types of radiative transitions.
Fluorescence: when a molecule in the (S1) state returns to the ground state (S0), emission of
radiation occurs in about 10-8 s. This process is known as Fluorescence.
Phosphorescence: when a molecule in the (T1) state returns to ground state (S0), emission Of
radiation occurs at a slow rate 10-4 s. This process is known as Phosphorescence.
 Fluorescence and Phosphorescence:
The two phenomena Fluorescence and Phosphorescence are also known as Chemiluminescence.
Fluorescence:
• Many compounds absorb UV light radiations and undergo electronic transition from lower
energy levels to higher energy levels.

• This absorption of light requires 10-15seconds. Then the energy absorbed will be re-emitted
within a short time of 10-8seconds as Fluorescence.

• The wavelength of fluorescence radiation is longer than absorbed radiation.

• Hence Fluorescence can be described as ' Instantaneous re-emission of absorbed light'.

• Emission of Fluorescence stops as soon as the incident exciting light radiation is cut off.

• It is shown by solids, liquids and in case of gases at low pressure.

Phosphorescence:
• Sometimes electrons in the excited state take more time to come to ground state by re-
emitting light radiation.

• In this case the re-emission of light takes place after 10-4seconds to several seconds.

• This delayed re-emission of absorbed light is called Phosphorescence.

• The phenomenon of Phosphorescence continues even after the exciting light radiation is cut
off.

• Phosphorescence is observed because of slow return of electrons to the ground state energy
level.

• Life time of Phosphorescence is much longer than Fluorescence.

• In Phosphorescence, the re-emitted light radiations are of low energy and longer wavelength
than Fluorescence.

• Phosphorescence is shown only by solids.

Distinguish between Fluorescence and Phosphorescence:

No Fluorescence No Phosphorescence

1 It is absorption of energy by atom or 1 It is absorption of energy by atom or

molecule followed by immediate emission molecule followed by delayed emission of
of light. light.

2 The excited atom has comparatively 2 The excited atom has comparatively
short lifetime of 10-8seconds. long lifetime of 10-4seconds.
3 Emission of Fluorescence stops as soon as 3 The phenomenon of Phosphorescence
the incident exciting light radiation is cut continues even after the exciting light
 off radiation is cut off.

4 It is shown by solids, liquids and in case of 4 It is shown only by solids.

gases at low pressure.

5 eg. CaF2 and Na, I, Hg vapours. 5 eg. ZnS, CaS and BaS etc.

Q. Applications of fluorescence in medicines

1. Laser induced fluorescence spectroscopy of human tissues for cancer diagnosis
A drug is systematically administered so that it is retained by a tumor. This drug show fluorescence
when light of appropriate wavelength is incident on it. This fluorescence is used for detection and
imaging of the tumor.
2. Detection of blood glucose Abnormal amount of glucose in human blood can be detected with
the help of fluorescence.
3. Photodynamic therapy (PDT) is a treatment that uses a drug, called a photosensitizer or
photosensitizing agent, and a particular type of light. When photosensitizers are exposed to a
specific wavelength of light, they produce a form of oxygen that kills nearby cells.
4. Intravascular fluorescence is a catheter-based medical imaging technique that uses fluorescence
to detect high-risk features of atherosclerosis (hardening of the arteries) and unhealed vascular
stent devices.
(Application of Spectroscopy 16M)
1. What is emission spectroscopy?
2. What is flame emission spectroscopy?
3. Discuss the basic principle/ theory of flame Photometry
4. Explain the instrumentation in flame photometer or With the help of a neat labelled
diagram, describe various components of flame photometer and their functions.
5. What are the advantages, disadvantages and applications of the flame photometry.
6. Write a note on fluorescence.
7. Write a note on phosphorescence.
8. Distinguish between fluorescence and phosphorescence.
9. Give the application of fluorescence in medicine.
10. Write a note on Jablonski diagram.
11. Explain calibration method for finding out the concentration of unknown sample in
flame photometry.
12. Explain Standard addition method for finding out the concentration of unknown
sample in flame photometry.