CH8491-INSTRUMENTAL METHODS OF ANALYSIS

1.What do you meant by dual property?

Each type of electro magnetic radiation ie. Radio waves, ultra violet infrared,
visible etc has both the properties of a wave as well as a particle this
property is called as dual properties.

2.What are the different type of energy levels?

1. Atomic energy levels.
2. Molecular electronic energy level
3. Vibrational energy level.
4. X-ray energy levels.

3.What do you meant by percent transmittance?

Percent transmittance is the percent of the intensity of the original radiation
which passes through the sample.

Intensity
%T= ×100
originalintensity

4.What do you meant by Vibrational energy levels?

Vibrational energy level are usually hence smaller magnitude than electronic
levels.In diatomic molecules the allowed Vibrational structuring of the bond
between two atoms. The allowed energy bend for Vibrational is given by

� 1 �h k
g+ �
Evits = � g = 0,1, 2
� 2 �2p u
m1m 2
k - force constant μ-reduced mars m =
m1 +m 2

5.What do you meant by electro magnetic radiation?

The wave nature of electro magnetic radiation can be represented by electrical
and magnetic vectors – Besides being perpendicular to each other. Two fields
are perpendicular to the direction propagation. It is because of there associated
electric and magnetic field that the radiation is called electro magnetic
radiation.
6.What do you meant by radiant energy?
Radiant energy may be defined as the energy, the propagation and transfer of which takes place
as a wave motion with out transfer of matter the term radiant energy is generally used in the
reference to electro magnetic radiation changing from the electric wave of low frequency
through UV rays, visible lays.

7.What is wave length?

The linear distance between successive maxima or minima of a wave is known as wave
length. It is usually expressed cm.

8.What are the fundamental components of spectrophotometer?

1. Radiation source
2. Filler or monochromatic.
3. A pair of converters.
4. Photo meter.
5. Detector
6. Test sample

9.What is frequency?
The number of complete cycles per second is called frequency. It is expressed in Hertz.

10.What is the wave length range for x-ray, UV, visible rays?

x-ray  0.01-15nm

for UV  15-200nm

visible  400-800nm.

BIG QUESTIONS

1.DESCRIBE ABOUT THE DUAL PROPERTIES AND RANGES OF THE

ELECTROMAGNETIC RADIATION.

Basic concepts:

Conduction: it occurs when one body (atom or molecule) collide with other

Convection: energy is transferred from one place to another by physically

moving the bodies

Radiation: it is the only form of energy transfer that can take place in vacuum
such as the region between the sun and the earth
 1. Electro magnetic radiation:

Electromagnetic radiation is the form of energy that is transmitted through

space with an enormous velocity and there by associated with both electrical
and magnetic property

2. Spectrum:

When a narrow beam of light is allowed to pass through a prism or grating, it is

dispersed into seven colours from red to violet and the set of colours or band
produced is called spectrum.

3. Radiation:

Radiation is a energy travelling through space as a series of waves or as a

stream of particle

4. Electromagnetic spectrum:

The arrangement obtained by arranging various types of Electromagnetic

radiation or waves in order of their in increasing wavelengths and decreasing
frequency is called Electromagnetic spectrum.

The spectrum by white light is called continuous spectrum.

Properties of Electromagnetic radiation:

 An Electromagnetic radiation is said to have a dual nature exhibiting

both wave and particle characteristics
 An Electromagnetic radiation is an alternating electrical and associated
magnetic force field in the space.
Wave properties of Electromagnetic radiation:

1. Wavelength:

 The distance between two successive maxima on an electromagnetic

wave.
  It is denoted by λ
 Unit of wavelength are
i. 1μ=10-6=10-4cm

ii. 1nm=10-9m=10-7cm
iii. 1Aº=10-10m=10-8cm

 A beam carrying radiation of only one wavelength is called

monochromatic
 A beam having radiation of several wavelength is said to be
polychromatic or heterochromatic
It should be noted that

(a) wavelength of visible light(3800 Aº-7600 Aº)determine its colour

(b) Wavelength of radiation is inversely related to its energy.

2. Frequency:

 The number of waves per second is called frequency of an

Electromagnetic radiation
 It is defined as the number of wavelength unit passing through a given
point per unit time
 It is denoted by ν
 It is generally expressed in cycles per second or hertz(Hz)
o 1 Hz =1cycle/ sec
o 1MHz=106cycle/sec

speed of lig h t ( c ) ∈cm/sec

Frequency( ν )=
waveleng h∈cms

The velocity of light c=3.0×1010cm/sec

 It should be noted that

(a) frequency of visible light determine its colour for example,violet

color corresponds to frequency of 7.89×1014cycles/sec

(b )frequency of radiation is direct measure of its energy.

3. Wave number:

 It is defined as the number of waves per centimeter in vacuum .

 It is denoted by V
 It is equal to the reciprocal of wavelength in centimeter
1
V=
λ

 Wave number has unit of reciprocal of distance ,Kaiser (k)

1k= cm-1

4. Velocity:

The distance travelled by the wave in one second is called velocity.

It is denoted by C

C=λ ν

Relation between wavelength and Partial properties of Electromagnetic

radiation:

The relationship between energy E of a radiation and its frequency (γ) is given
by Planck
E
E=h ν h-plancks constant,h=
ν

Where, h-Planck’s constant=6.626 ×10−27 ergs/sec

Relation between wavelength and Partial properties of Electromagnetic

radiation:

E=h ν
C
( Since C=λ ν :ν = )
λ
C
E=h
λ
 1
(since V = )
λ

E=h c v

It should be noted that frequency is directly proportional to energy, but

energy increases, the wavelength decreases

Important characteristics of the electromagnetic radiation

 These are produced by the oscillation of electric charge and magnetic

field residing on the atom . The electric and magnetic components are
mutually perpendicular to each other and are coplanar.

 These are characterized by their wavelength of frequency of wave

number.

 The energy carried by the 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 type of radiations travel with the same velocity and no medium is
required for their propagation. They can travel through vacuum.

 When visible light (a group of electromagnetic radiation ) is passed

through a prism, it is split up into seven colors which correspond to
define wavelength to definite wavelength. This phenomenon is called
dispersion.
 Electromagnetic Radiation is basically light, which is present in a
rainbow or a double rainbow.
 It also is a spectrum consisting of radio waves, microwaves, infrared
waves, visible light, ultraviolet radiation, X-rays, and gamma rays. There
are only two ways to transfer energy from one place to another place.
 Through a particle.
 Through a wave.
 An electromagnetic radiation is a particle as well as a wave thus it is
interesting to study its nature in quantum theory. Light is also an
electromagnetic radiation which contains frequencies.
 The photoelectric effect could be explained considering that radiations
consist of small packets of energy called quanta.
 These packets of energy can be treated as particles. On the other hand,
radiations exhibit a phenomenon of interference and diffraction which
indicated that they possess wave nature.
 So it may be concluded that electromagnetic radiations possess dual
nature. Particle nature and Wave nature
Particle Nature of Electromagnetic Radiation

Planck’s Quantum Theory – It states, “One photon of light carries exactly

one quantum of energy.” Wave theory of radiation cannot explain the
phenomena of the photoelectric effect and black body radiation.

• What is Black Body Radiation?

An ideal black body is a perfect absorber and perfect emitter of radiation.

When such a body is heated, it becomes red hot.

Wave Nature of Electromagnetic Radiation

• Due to a dispersion of white light, VIBGYOR appears. Vibgyor stands for

– Violet, Indigo, Blue, Green, Yellow, Orange, and Red.

2. DESCRIBE THE ATOMIC ENERGY LEVELS.

Atomic energy is the quantum theory, atoms can exist only at dispute potential energy level.
The potential energy of an atom depends on the electron conjugation and transition of outer
relation between fixed energy level to emit an absorb radiation at discrete energies. The
frequency of radiation absolved or emitted is proportional to the change is potential energy
involved. The atomic sprite involves only transition of electrons from one electronic energy
level another. Each transition amounts for the presence of a specific light frequency and the
spectral either in absorption or emission .Atomic band, diagram for Na is guest below

A special emission line results from a transition. From a higher energy band to lower one-
the energy transition is from ground state to excited state. Only a few atoms with labile electron
have absorption spectra in the visible region when an atom ins executed by absorbing light or
by collision with excited directions, the excited Loris or molecules remains in an executed state
for only a very short time, approximately in 10-9 sec.
The major contribution to the energy terms are associated with a principal quantum
number n=1,2,3……..
For Ex. When 3s valence electron of sodium is excited to 4s, 5s, 6s and larger orbital. The series
of energy levels labelled 2s, the separation the consolation level states at n= .
 The resultant orbital angular momentum or resultant essentiality of electronic orbital
having the same principal quantum number accounts for smaller energy difference which are
classified as different series.

The resultant angular momentum is the vector sum of the oriental angular moments of
the electronic orbital having the same principal quantum numbers accounts for smaller energy
differences which are classified as different series. The resultant angular momentum is the
vector sum of orbital angular momentum of the individual electron.
L=0,1,2,3………
Represented by the quantum number L=0,1,2,3…….(term symbol s,p,D,f…….) for example a 2p
series results when the single volume of electron of sodium is executed to p status of higher
orbit.

3. DERIVE THE EXPRESSION FOR BEER’S LAW .(OR) WHAT DO YOU

MEANT BY BEER-LAMBERTS LAW? EXPLAIN IT.
4. DISCUSS IN DETAIL ABOUT REGIONS OF ELECTROMAGNETIC
RADIATION.
Regions of Electromagnetic radiation:
  Visible light is a form of electromagnetic radiation which lies in the wavelength range of
3800 Aº-7600 Aº
 The region of 3800 Aº wavelength corresponds to violet colour
 The region of 7600 Aº wavelength corresponds to red colour
 If the wavelength is less than 3800 Aº, the radiation is called ultraviolet light
 If the wavelength is greater than 7600 Aº, the radiation is called infrared light
 Infrared as well as ultraviolet radiation are not visible to eyes and so these are known as
invisible radiations

Regions of Electromagnetic radiation are

1. Radio wave Region

2. Microwave Region
3. Infrared Region
4. Visible Region
5. Ultraviolet Region
6. X-ray Region
7. γ -ray Region
8. Cosmic ray Region
5. EXPLAIN THE INTERACTION OF ELECTROMAGNETIC WAVE WITH
MATTER

Spectroscopy:
Spectroscopy is the branch of science dealing with the study of
interaction of electromagnetic radiation with matter.
The study of spectroscopy carried out under
1. Atomic spectroscopy
2. Molecular spectroscopy
1. Atomic spectroscopy:
This Spectroscopy is concern with the interaction of electromagnetic radiation
with atoms
Which are commonly in their lowest energy state, called the ground state.
Atomic spectra involve only transition of electrons from one electronic level to
another.

2. Molecular spectroscopy:
This Spectroscopy is concern with the interaction of electromagnetic radiation
with molecules.
Molecular spectra involve transition between rotational and vibrational energy
levels, in addition to electronic transition.
lowest energy state is called ground state; other states are called exited state.

1.ABSORPTION OF RADIATION:
When electromagnetic radiation passes through
matter, a variety of phenomena may occur. Some of these are as follows:
CASE1: Fully absorbed and emitted instantly
CASE2: partially absorbed
CASE3: scattering and reflection
CASE4: absorbed and emitted later

2. ABSORPTION AND EMISSION SPECTRA:

Spectroscopy is mainly concerned with the interaction of electromagnetic
radiations with matter. After interaction, there may occur variation in intensity
of electromagnetic radiation with frequency or wavelength. The instrument
which records this variation in intensity of radiation is known as spectrometer.
There are two ways in which the interactions are observed,
 EMISSION SPECTRUM:
Sample emits radiation itself.
When a solid is heated to a high temperature or a discharge
of electricity is passed through a gas ,light is emitted .on
examining this light by spectroscope, a spectrum has been
found to be produced which is known as emission spectrum.
 spectrometer

sample
exited thermally
emission frequency
or electrically spectrum

 ABSORPTION SPECTRUM:
Sample absorbs radiations from a continuous source.

When white light is passed through a coloured solution or

vapour, and the spectrum obtained is examined through a
spectroscope, black bands are noticed, because light of
wavelength corresponding to black bands is absorbed by the
substance through which the light is passed. The spectrum
so obtained is called absorption spectrum.

spectrometer

Source continuous radiation sample

absorption frequency spectrum

Spectra can be divided into two types depending on the spectral lines:

• Line Spectrum or Atomic Spectrum – This is made up of distinct lines.

When an electron in an atom excites and de-excites, this spectrum
occurs. Emission & absorption spectra show the line spectrum.
 • Band Spectrum – It is a characteristic of a molecule. It consists of
closely spaced lines called bands. In a molecule, the vibration & rotation
of atoms generates such spectrum

3. ATOMIC ABSORPTION OF ELECTROMAGNETIC RADIATION:

 When an atom absorb electromagnetic radiation, this result in an
increase in their electronic energy.
 The absorption can only occurs if the attracting photon of
electromagnetic radiation possesses energy which is equal to the energy
difference between two quantized energy level of the absorbing atom.
 The increase in electronic energy results as the atom jumped in the
excited state. When the excited atom reverts to the ground state, it loses
energy in one straight jump or through a number of steps in the form of
a single quantum or photon of electromagnetic radiation.
Some silent features of the line spectra are:
 The spectral lines are regularly spaced.
 The wavelengths of the lines emitted are the
characteristics of the elements under consideration.
 Line spectrum is independent of the compound in
which the element may occur.
4. MOLECULAR ABSORPTION:
When the molecule absorbs electromagnetic radiation, it gets excited and
when it returns to the ground state, it emits band which are
characteristics of the molecule concerned and that is why we call this as
molecular spectra.
BAND HEAD: If the absorbed the line the wavelength edge, the lines
become closer and closed until they coincide. This edge is known as band
head.
TYPE OF ENERGY LEVEL:
 Rotational energy
 Vibrational energy
 Electronic energy
5. REFRACTION AND DISPERSION:
REFRACTION:
When a beam of light is allowed to pass from one medium to another having a
different physical density than the first, an abrupt change in the direction of
the beam is observed. This is due to the difference in the velocity of radiation in
two media. This phenomenon is called refraction
The refractive index of a medium is the ratio of velocity of light
in vacuum to its velocity in the medium.

n=c/v
where ,
n= refractive index
 c= velocity of light in vacuum
v= velocity of light in medium
The change in refractive index will also change in wavelength. Therefore the
velocity is depends upon the wavelength of radiation.
The refractive index will also depend upon the composition of the matter
through which passing.

DISPERSION:
The change in refractive index which occurs with a change
in the wavelength of transmitted light is called dispersion.

6. SCATTERING AND REFLECTION:

SCATTERING:
Nephelometry and turbidimetry are based upon this ability of particles to
scatter light. The size, shape, concentration of colloidal particles may be
determined from this property.

REFLECTION:
Reflection takes places when beam of radiation is allowed to cross an interface
between media of different refractive index.
It is dependent upon the refractive index of the two media and increase at
these differences in the refractive index increases.

6. DISCUSS THE QUANTITATIVE ANALYSIS OF ANY ONE ORGANIC SALT

USING DUBOSQ COLORIMETER.

PRINCIPLE:
The basic principle of Duboscq colorimeter is based on Beer’s Law
Duboscq colorimeter estimates the length of light that can travel through a
solution. The concentration is proportional to the number of color matches and
wavelength of color. The unknown concentration of solution can be easily
determined by colorimeter.

WORKING:

1. Colorimeters are used to determine the concentrations of substances by

measuring color intensities.
2. The DuBoscq type of colorimeter works by comparing the colors of two
solutions of a colored substance,
3. a standard with a known concentration, and a sample with an unknown
concentration.
4. The known solution is taken into a cylinder or a tube with a flat
transparent base
 5. The standard solution is taken into a second cylinder
6. A plunger is kept in each cylinder. the plungers moved up and down
until the colors seen from the top of each cylinder become identical
7. From the reading of the depth of the sample, the concentration of the
unknown can be evaluated

(concentration 1)(pathlength 1) = (concentration 2)(pathlength 2)

b1c1 = b2c2

(concentration 2)= (concentration 1)(pathlength 1)/ (pathlength 2)

C 1×b1 C2=
b2

ADVANTAGES
 It is economical, fast and has the simple operation of a spectrometer.
 It is a fast and convenient method as compared with the volumetric or
gravimetric processes and they are easily optimized for automation.
 It does not require an experienced person to handle it.
 The chemical substances in water can be identified by this method.
 It applied to the quantitative analysis of colored compounds.
 Another advantage of colorimetry is that it is a portable system you can
easily carry and transport.
LIMITATIONS
 The major disadvantage of colorimetry is that colorless compounds
cannot be analyzed.
 It needs more amount of sample for analysis.
 You require preparing a standard solution.
 Its sensitivity is low.
 The same colors from interfering material may create errors in results.
 The precise wavelength bandwidth may be required for more accurate
analysis.
 The interference with the matrix can lead to poor results in uncontrolled
conditions.
APPLICATIONS
 The colorimeter is a device used to determine the colors based on the
transmittance and absorption of light. A colorimeter can measure the
concentration of a substance which is depending on the intensity of the
color of the in the sample solution.

 The colorimeter has a wide scope of applications is used for quantitative

and qualitative analysis of colored compounds, including determining the
concentration of a solution, laboratory research, the analysis of blood,
environmental analysis of water quality, determining the rates of
reaction, applications across the chemical and biological fields, analysis
of soil components, nutrients in soil and foodstuffs, analysis of chemicals
used indifferent industrial settings.
 Some other applications of colorimeter the analysis is used in the food
ingredients, textile products, building materials, beverages, chemical
solutions, and many others.

7. DESCRIBE ABOUT MOLECULAR ELECTRONIC ENERGY LEVEL.

The molecular electronic energy are represented by potential energy curves. Hence the P.E.
of electronic state is plotted against inter nuclear distance.

An electronic energy level is send to he stable. When the P.E is minimum the molecular
how executed electronic state.

According to the pauli exclusion principle spin of buy electron in the same orbital are opposite
to one another that is period.

When the molecules have even number of electron in the same orbital. They are
arranged in opposite direction.
 Be fore the molecule absorb radiation they must in heart. The electron with more rapidly
than nuclear. So the nuclear do not change their position. This principle is given by frank-
condom.

The above energy level. Diagram shows the electronic transitions in absorption, shows a
series of lovely spaced lines an responding to a different vibration levels.

i. The molecule absorbs radiation and it go to executing state. The excited molecule can
come hence to the lower energy level by several path.
ii. The molecules can come been to lower energy by irrational relaxation.
iii. When molecules reaches lowest single excited state, the electrons are return to low
ground electronic state so, this is called florescence.
iv. If the P.E came of the excited singlet creases that of triplet state. Some of executed
molecules pars when triplet state is a an inter system crossing, which involves coupling
between excited surgical state (T2) and triplet state( T1) other the triplet state is expressed
the molecules under go Vibrational relaxation to arise the lowest excited triplet state.
8. DESCRIBE ABOUT THE INSTRUMENTS USED TO MEASURE THE
TRANSMITTANCE OR OPTICAL DENSITY.

(1) SPECTROPHOTOMETER

DIAGRAM

PRINCIPLE OF SPECTROPHOTOMETER

Spectrophotometer is based on the photometric technique which states that

When a beam of incident light of intensity I 0 passes through a solution, a part
of the incident light is reflected (Ir), a part is absorbed (Ia) and rest of the light is
transmitted (It)

Thus,

I0 = Ir + Ia + It

⇒ In photometers (colorimeter & spectrophotometer), (Ir) is eliminated because

the measurement of (I0) and It is sufficient to determine the (I a). For this
purpose, the amount of light reflected (I r) is kept constant by using cells that
have identical properties. (I0) & (It) is then measured.

⇒ The mathematical relationship between the amount of light absorbed and

the concentration of the substance can be shown by the two fundamental laws
of photometry on which the Spectrophotometer is based.

TYPES OF SPECTROPHOTOMETER

Spectrophotometer is of 2 types –

 Single beam spectrophotometer

  Double beam spectrophotometer

Single beam spectrophotometer operates between 325 nm to 1000 nm

wavelength using the single beam of light. The light travels in one direction and
the test solution and blank are read in the same.

Double beam spectrophotometer operates between 185 nm to 1000 nm

wavelength. It has two photocells. This instrument splits the light from the
Monochromator into two beams. One beam is used for reference and the other
for sample reading. It eliminates the error which occurs due to fluctuations in
the light output and the sensitivity of the detector.

PARTS OF SPECTROPHOTOMETER

There are 7 essential parts of a spectrophotometer

Light source – In spectrophotometer three different sources of light are

commonly used to produce light of different wavelength. The most common
source of light used in the spectrophotometer for the visible spectrum is a
tungsten lamp. For Ultraviolet radiation, commonly used sources of are the
hydrogen lamp and the deuterium lamp. Nernst filament or globar is the most
satisfactory sources of IR (Infrared) radiation.

Monochromator – To select the particular wavelength, prism or diffraction

grating is used to split the light from the light source.

Sample holder – Test tube or Cuvettes are used to hold the colored solutions.
They are made up of glass at a visible wavelength.

Beam splitter – It is present only in double beam spectrophotometer. It is used

to split the single beam of light coming from the light source into two beams.

Mirror – It is also present only and double beam spectrophotometer. It is used

to the right direction to the splitted light from the beam splitter.

Photodetector system – When light falls on the detector system, an electric

current is generated that reflects the galvanometer reading.

Measuring device – The current from the detector is fed to the measuring
device – the galvanometer. The meter reading is directly proportional to the
intensity of light.

WORKING OF THE SPECTROPHOTOMETER

⇒ When using a Spectrophotometer, it requires being calibrated first which is

done by using the standard solutions of the known concentration of the solute
that has to be determined in the test solution. For this, the standard solutions
are filled in the Cuvettes and placed in the Cuvette holder in the
spectrophotometer that is similar to the colorimeter.

⇒ There is a ray of light with a certain wavelength that is specific for the assay
is directed towards the solution. Before reaching the solution the ray of light
passes through a series of the diffraction grating, prism, and mirrors. These
mirrors are used for navigation of the light in the spectrophotometer and the
prism splits the beam of light into different wavelength and the diffraction
grating allows the required wavelength to pass through it and reaches the
Cuvette containing the standard or Test solutions. It analyzes the reflected
light and compares with a predetermined standard solution.

⇒ When the monochromatic light (light of one wavelength) reaches the Cuvette
some of the light is reflected, some part of the light is absorbed by the solution
and the remaining part is transmitted through the solution which falls on the
photodetector system. The photodetector system measures the intensity of
transmitted light and converts it into the electrical signals that are sent to the
galvanometer.

⇒ The galvanometer measures the electrical signals and displays it in the

digital form. That digital representation of the electrical signals is the
absorbance or optical density of the solution analyzed.

⇒ If the absorption of the solution is higher than there will be more light
absorbed by the solution and if the absorption of the solution is low then more
lights will be transmitted through the solution which affects the galvanometer
reading and corresponds to the concentration of the solute in the solution. By
putting all the values in the formula given in the below section one can easily
determine the concentration of the solution.

⇒ In double beam spectrophotometers, the beam splitters are present which

splits the monochromatic light into two beams one for the standard solution
and the other for test solution. In this, the absorbance of Standard and the
Test solution can be measured at the same time and any no. of test solutions
can be analyzed against one standard. It gives more accurate and precise
results, eliminates the errors which occur due to the fluctuations in the light
output and the sensitivity of the detector.

APPLICATIONS OF THE SPECTROPHOTOMETER

⇒ The spectrophotometer is commonly used for the determination of the

concentration of colored as well as colorless compounds by measuring the
optical density or its absorbance.
⇒ It can also be used for the determination of the course of the reaction by
measuring the rate of formation and disappearance of the light absorbing
compound in the range of the visible & UV region of electromagnetic spectrum.

⇒ By spectrophotometer, a compound can be identified by determining the

absorption spectrum in the visible region of the light spectrum as well as the
UV region of the electromagnetic spectrum.

⇒Here is the formula used for determining the Concentration of a

substance in the Test solution using the absorbance values measured by
the spectrophotometer….

ADVANTAGES AND LIMITATIONS

The advantages of the single beam design are low cost, high throughput, and
hence high Sensitivity , because the optical system is simple.
The disadvantage is that an appreciable amount of Time elapses between
taking the reference (I) and Making the sample measurement (Io) so that there
can be problems with drift

(ii) PHOTOELECTRIC COLORIMETER

DIAGRAM
The field of chemistry is full of devices that are used for different tests and
measurements. Operating these devices take some good technique and
learning. Colorimeter is one of those devices in the field of chemistry. The light
waves of various wavelengths can be absorbed different solutions. However,
before this device, we didn’t know exactly how to calculate that. Now, with the
Photoelectric Colorimeter, we can easily measure the ability of any solution to
absorb the light waves of various wavelengths. This device has helped
scientists in a lot of ways. Many scientific experiments have been made easy
with the help of this device.

Photoelectric Colorimeter Definition:

Photoelectric colorimeter is a device that is used for the quantitative analysis of

different substances like glucose, blood urea, blood enzymes etc. It works on
the principle of absorption of light which is explained in detail below.

Photoelectric Colorimeter Working Principle:

Most of the students learning advanced sciences know about the Beer-
Lambert’s Law. According to this law, the medium concentration of any liquid
is directly related to the waves absorbed in the liquid when waves are passed
through it. This device is based on the same principle and you will need be
understanding in order to get it. You can only perform and understand this
experiment well if you know about Beer Lambert’s Law.

How to use Photoelectric Colorimeter?

Here is an easy step by step guide to show you how this device works. There is
nothing complex in the principle and working of the colorimeter. However, if
you are not familiar with the Beer-Lambert’s Law, then you must read it first.
The working of this device is based on it and you can do better if you
understand the experiment. Let us tell you with a step by step guide to teach
you how to use Photoelectric Colorimeter.

1. The whole operation is carried out with the help of few lenses,
colorimeter, and a light source. There are measurement recorders in the
colorimeter as well.

2. First, you pass the light waves through the fluid. Make sure with the
help of lenses that these waves reach the measuring device.

3. Once these waves who have the same wavelengths pass through the
measuring meter, a reading is recorded.
 4. Different readings are taken by repeating the same process.

5. Then the readings are compared to see the final results of the ability of
the fluid to absorb these light waves.

And, this is how you easily perform this experiment with the help of
Photoelectric Colorimeter.

Photoelectric Colorimeter Applications:

· This device can be used to do a number of things. First, this device can be
used to measure the growth rate of yeast or bacterial material. The device is
known for its higher accuracy to do the thing. Also, in the bird’s plumage, you
can find the assessment of colors with the help of this device.

· And this is not all. Photoelectric Colorimeter is not just used for the lab
purposes in the field of chemistry. The quality of water can also be measured
with the help of this device. Different chemicals and their qualities are tested
simply by using this amazing measuring device.

ADVANTAGES

Colorimeters are extraordinarily accurate for straightforward color

measurement and ideally suited for determination of color difference, fastness,
and strength as well as routine comparisons of similar colors. As such, they
can be invaluable for color quality control and are primarily used in the
production and inspection phases of manufacturing.

DISADVANTAGES

While colorimeters can produce highly accurate color measurements, they also
have several shortcomings; they are not able to identify metamerism or
colorant strength, are not ideally suited for color formulation, and cannot be
used under variable illuminant/observer conditions.
 9. CLASSIFY THE INSTRUMENTS DEPENDS ON THE METHOD PHYSICAL
PROPERTIES

Method Principle Property measured

1. Electro
Change in the electrical properties of
chemical Mass of deposited substance
the system.
method.

Material deposited
a. Electro
Electrolysis is carried are. represented on one of the
gravimetry
electrode is weighed.

Deposition of matter on an electrode

b. Coulometry Quantity of electricity
during electrilyrs

Change in electrical conductivity of a Electrical conductivity,

c. Conduct metry
solution duty chemical electrical resistance.

Potential applied between the

D. Amperometry
indicator

Electrode and depolarized reference Current through the

e.Electrolytic cell
electrode is dept constant electrolytic cell is measured.

2.optimal
method

1. atomic Anomissing the specimen Absorption of radiation

thsorption
spectroscopy

2.Emission Position and intensities of

Sample is subjected to an electric are
spectroscopy special lines

3. X-ray Position and intensity of

Emission of X-ray spectrum atom
spectroscopy special lines

Absorption and setting of a light beam

4. Turbridimetry Same
by turbind media
 Reflection and scattering of a light
5. Nephelometry Amount of light stopped.
beam by collladal solution.

Ionisation of atoms , ions and Position and intensity of

3. Mass
moleculers by a combined action of signals in mans spectrum.
spectroscopy
electric and magnetic fields. Mass to change ratio.

4. nuclear
Postion and incentivity of
magnetic Nuclear magnetism
lines of NMR spectrum.
resonance

5. Radio metric Concession by sibale is o topes of an Intensity of radiation

methods element to radio is topes induced radio activity.

Concomitant change in the

6. Kinetic Speed of chemical reaction may be in
absorbance of solution for
method, measured by the addition of catalys.
visible or UV radiation.

7. thermal Recording as a function of

Change in weight or energy.
methods: temperature and time

9. EXPLAIN THE WAVE PROPERTIES OF ELECTROMAGNETIC

RADIATION AND ALSO DISCUSS IN DETAIL ABOUT ELECTROMAGNETIC
SPECTRUM.

ANSWER:

REFER QUESTION NO 1 AND QUESTION NO 4

10. HOW DO YOU ESTIMATE IRON IONS QUANTITATIVELY USING BEER

LAMBERT LAW

STANDARTIZATION OF PERMANGANATE SOLUTION

• Take 10 ml of sodium oxalate solution (Na2C2O4) and add 90 ml of H2SO4

solution.
• Heat the solution nearly to boiling.
• Solution was titrated with permanganate solution until a pink coloration
appears . Keeping the solution above 70oC throughout the titration.
• Then from the corrected volume of permanganate , calculate the molarity
of the permanganate solution .
 •Then from the corrected volume of permanganate , calculate the molarity
of the permanganate solution .
PREPARATION OF SAMPLE :
• To prepare a solution which is 1.0 N in iron (III) chloride (FeCl3), add 4.0
g FeCl3 to 25 mL DI water. Then add 8 mL concentrated hydrochloric
acid to the solution.

IRON DETERMINATION:
• 5 ml of sample was taken.
• 20 ml of distilled water was added.
• Solution heated on the bunsen burner.
• Stannous chloride (SnCl₂) was added until the yellow color changes into
light green. A few excess drop was added.
• Solution was diluted to 100 ml and cooled.
• 10 ml of mercuric chloride, HgCl2, was added. Precipitation was
observed. It was passed to next step in case precipitation was white; else
previous steps were repeated with a new sample.
• Solution diluted to 300 ml with distilled water and 25 ml of
Zimmermann-Reinhardt reagent was added.
• Solution was titrated with permanganate solution.
• Calculate the percentage Fe in the sample