Sub: Engineering Chemistry

Module 5: Instrumental methods of analysis

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 MODULE-V

INSTRUMENTAL METHODS OF ANALYSIS

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 INTRODUCTION

Chemical analysis: The study of the chemical composition and

structure of substances.
Chemical analysis

Qualitative analysis Quantitative analysis

Identification of chemical Amount of each constituent
constituents(What ?) ( How much?)
( Salt and compound analysis)
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 Quantitative analysis
a) Classical method( Wet Chemical method) b) Instrumental analysis

Classification of Wet Chemical method

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 Classification of Instrumental Methods of analysis
Advantages of instrumental methods:-
Instrumental Analysis 1.A small amount of a sample is needed
for analysis.
2.Determination by instrumental method is
considerably fast.
Electrochemical Spectroscopic 3.Complex mixture can be analyzed either
Photometry
Method Method with or without their separation.
4.Sufficient reliability and accuracy of
Conductometry Colourimetry Flame photometry results are obtained by instrumental
method.
5.When non-instrumental method is not
Potentiometry Atomic Absorption possible, instrumental method is the only
Spectroscopy
answer to the problem.

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 Colorimetry
• Colorimetry is a scientific technique used to determine the concentration of
colored compounds in solutions by the application of the Beer–Lambert law. 
 

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 PRINCIPLE

When a copper solution is treated with ammonium hydroxide, it

forms a deep blue colured cuprammonium complex. Intensity
of the colour increases with increase in concentration of copper
ions. The absorbance of the blue colour solution is measured at
620 nm. Cu 2  4NH   [Cu(NH ) ]2
3 3 4
Cupraammonium complex

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 INSTRUMENTATION

Source: Tungsten bulb is used as a light source.

Filter: It is a device for isolating monochromatic light.
Sample: Glass or quartz cuvette is used as sample holder.
Photocell: Converts the emitted light into electrical signal.
 
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 Working

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 Application of Colourimetry
Determination of copper in brass sample
Procedure:
 Pipette out 2, 4, 6, 8, 10ml of standard copper sulphate solution Absorbance
into 25ml standard flask.
 Add 2.5ml of ammonia solution into each of them and make up
to the mark with distilled water and shake well.
 Blank solution is prepared without copper solution as above.
 A known quantity of unknown solution is taken in a standard
flask and step 2 is followed.
 Set the filter to 620nm, adjust the initial reading to zero by using Concentration of Copper Sulphate

blank solution in the sample tube.

 Measure the absorbance for each standard flask solution and plot
graph of absorbance v/s concentration of copper sulphate.

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 Application of Colourimetry

Applications
 In quantitative analysis: large number of metal ions, anions and cations compounds can be determined by in this
method
 Determination of the composition of colored complex.
 
Advantages Disadvantages
The method is much faster than the chemical methods. Technique can not be used for colourless analyte.
The analysis can be conducted in a very short time.

It requires small quantities of the analyte. Technique is applicable only in visible region.

They give accurate results. Interference by same colour substances.

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 Flame Photometry
A method of determination of metal ion concentration by measuring emission intensity is called flame
photometry. Metal ion concentration is directly proportional to emission intensity.

Principle:

The principle involved in flame photometry is that when a solution containing a metallic
compound is aspirated into a flame, vapours containing metal atoms will be formed. Some
of these metal atoms in gaseous state may be raised to an energy level which is sufficiently
high to permit the emission of radiation, which is characteristic to the metal under
investigation. This method is generally used for the analysis of sodium, potassium, calcium
and lithium.

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 DIAGRAM

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 Instrumentation

 Source of flame: A Burner in the flame photometer is the source of flame. It can be
maintained in at a constant temperature. The temperature of the flame is one of the critical
factors in flame photometry.
 Nebuliser:  Nebuliser is used to send homogeneous solution into the flame at a balanced rate.
 Optical system: The optical system consists of convex mirror and convex lens. The convex
mirror transmits the light emitted from the atoms. Convex mirror also helps to focus the
emissions to the lens. The lens helps to focus the light on a point or slit.
 Simple colour filters: The reflections from the mirror pass through the slit and reach the
filters. Filters will isolate the wavelength to be measured from that of irrelevant emissions.
 Photo-detector: The intensity of radiation emitted by the flame is measured by photo
detector. Here the emitted radiation is converted to an electrical signal with the help of photo
detector. These electrical signals are directly proportional to the intensity of light.
 

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 Working
Flame photometry employs a variety of fuels mainly air, oxygen or
nitrous oxide (N2O) as oxidant. The temperature of the flame depends on
fuel-oxidant ratio.
 The various processes in the flame are discussed below:
 Desolvation: The metal particles in the flame are dehydrated by the
flame and hence the solvent is evaporated.
Vaporization: The metal particles in the sample are dehydrated. This
also led to the evaporation of the solvent.
Atomization: Reduction of metal ions in the solvent to metal atoms by
the flame heat.

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Excitation: The electrostatic force of attraction between the electrons and
nucleus of the atom helps them to absorb a particular amount of energy. The
atoms then jump to the exited energy state.
Emission process: Since the higher energy state is unstable the atoms jump
back to the stable low energy state with the emission of energy in the form of
radiation of characteristic wavelength, which is measured by the photo detector.
The intensity of emitted light is directly related to the concentration of the
sample.
The comparison of emission intensities of unknown samples to either that of
standard solutions (plotting calibration curve), helps in the quantitative
analysis of the analyte metal in the sample solution.

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 Applications

To estimate sodium, potassium, calcium and lithium etc in sample of

serum, urine and other body fluids.
Flame photometer can be applied both for quantitative and qualitative
analysis of elements.
It is useful for the determination of alkali and alkaline earth metals.
Used in the determination of lead in petrol.
Used in determination of magnesium and calcium in cement.

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 Determination of sodium in water sample using flame photometer

1.Prepare the following standard solutions: 0.2, 0.4, 0.6, 0.8 and 1.0mg Na/100ml, from the 1000mg/l
sodium standard solution, using deionized water as the diluent.
2. Select the sodium filter and aspirate the 1.0mg Na/100ml standard and adjust sensitivity control to obtain
a reading of 100.
3.Aspirate deionized water and adjusts the zero control to obtain zero reading.
4. Aspirate 1.0mg Na/100ml standard again, re-adjust to 100.
5. Aspirate deionized water again and re-adjusts if necessary to zero.
6.Repeat steps 4 and 5 if necessary to obtain 100 on the standard and zero on deionized water.
7. Aspirate the other standard solutions, note the readings and plot a calibration curve.
8.Measure the emission intensity for water sample and obtain the concentration from the graph as shown
5.4.
9.Repeat the above calibration procedure with potassium filter, using standard solutions of the following
concentrations: 0.2, 0.4, 0.6, 0.8 and 1.0mg K/100ml, prepared from the 1000mg/l potassium standard
solution using deionized water as the diluent.

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 Determination of sodium in water sample using flame photometer

OTHER APPLICATIONS:
Flame photometer has both quantitative and qualitative
applications. Flame photometer with monochromators emits
radiations of characteristic wavelengths which help to detect the 100

presence of a particular metal in the sample.

Emission int
80
Calibration curve
This help to determine the availability of alkali and alkaline earth 60

metals which are critical for soil cultivation. 40

In agriculture, the fertilizer requirement of the soil is analyzed by 20 Unknown Conc

flame test analysis of the soil.
In clinical field, Na+ and K+ ions in body fluids, muscles and heart 0.2 0.4 0.6 0.8 1.0
can be determined by diluting the blood serum and aspiration into
Conc of Na+
the flame.
Analysis of soft drinks, fruit juices and alcoholic beverages can
also be analyzed by using flame photometry.

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 Advantages of flame photometer

Advantages Disadvantages
1. Applicable for both quantitative and 1. Technique is applicable only for alkali
qualitative analysis. and alkaline earth metals.
2.Microgram and milligram quantity 2. It is an emission spectroscopy.
analyte is sufficient for analysis. 3. Applicable for liquid samples.
3.More accurate results can be obtained
for low concentrations,

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 ATOMIC ABSORPTION SPECTROSCOPY
Definition: Atomic absorption spectroscopy is a technique which studies absorption
of electromagnetic radiations in relationship to molecular structure. It is a technique
for measuring the concentration of various elements in the sample through   their
absorption of light. 

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 Principle: When a solution having a mixture of metallic
species is introduced into the flame, first solvent evaporates
and vapor of metallic species is obtained. When a
Metal ions are atomized in the fame to produce gaseous ground monochromatic light of specific wave length light is passed
through the vapourized atoms present in the flame, then
state atoms. These atoms are excited by absorbing photons from atoms absorb the radiation and decrease in the intensity of the
the cathode lamp. Number of photons absorbed is proportional radiation will be directly proportional to the atoms present in
to ground state gaseous atoms and follows Beer-Lambert’s law. the ground state.
I0
A  log  Ct
It
Io= Intensity of absorbed light, It = Intensity of transmitted
light, ε = molar extinction co-efficient and t = path length.

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 2. Atomizer: - In which sample is burned with the flame
produced by the fuel & oxidant combination.
1. Nebulizer: -it creates a fine spray of the sample for
the introduction in the flame. In which the aerosol
& fuel & oxidant are mixed thoroughly for the
3.Light source: - The light source should produce a narrow
introduction into the flame.
spectrum of light. Generally Hallow-Cathode lamp used as light
source.

4. Monochromators:- it select a given

5. Detector: - it detects the absorption intensity.
wavelength of radiation from the source,
corresponding the energy gap b/w the exited
state & ground stat energy levels of the
analyte atom. 6. Recorder: LED screen to display reading

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 Working procedure
The metal salt solution is aspirated into the flame, first solvent evaporated
the tiny metal salt solute particles are obtained.
Further, the solute particles are converted into gaseous molecules at very
1.0 high temperature and then the molecules are dissociated into atoms.
Some of the atoms are excited and the excited atoms will absorb their
Sample absorption

0.75 Reading
Calibration curve
characteristic radiation. AAS is concerned with this part of light which are
0.5 absorbed by the excited atoms. The greater the number of atoms, the more
Sample concentration
radiation is absorbed. The amount of light absorbed is proportional to the
0.25
number of atoms.
0 2 4 6 8 10 12 14 16 18 20
A calibration curve is constructed by running several samples of known
concentration under the same condition as the unknown.
The amount of standard absorbance is compared with the calibration
Concentration of Nickel in ppm

curve and enables the calculation as the metal concentration in the

unknown sample.

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 Determination of nickel in steel sample using AAS

 Preparation of standard nickel solution: 0.1g of 100% nickel metal is dissolved in 2ml of HCl and made up to1000ml to
yield 100ppm nickel standard solution.( stock solution)

 Different aliquots of above solution is transferred into different 100ml standard flask and made up to mark with distilled
water to obtain 2,4.6,8 and 10ppm standard nickel solution.

 A blank is prepared only with 2 ml HCl and distilled water in 100ml flask.

 Sample solution is prepared as mentioned in first step and diluted to required volume using distilled water

 Nickel Hollow cathode lamp is placed in AAS instrument and wave length is adjusted to 232nm.

 Standard solutions are aspirated through Nebuliser and absorbance is recorded.( Capillary is cleaned with distilled water
after each aspiration)

 Aspirate sample solution and record its absorbance.

Plot a graph of absorbance Vs concentration to obtain calibration curve also, find the concentration of unknown sample

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Other Applications of AAS
 Atomic absorption spectroscopy is one of the most widely used techniques for the
determination of metals at trace levels in solution.
 It is more advantageous than flame photometry due to its relative freedom from
interferences
by inter-element effects and its relative intensity to various inflame temperature.

 Water analysis ( Eg; Ca, Mg. Fe, Si and Al)

 Food analysis, analysis of soils and clinical analysis.
 

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 Advantages of AAS
It does not suffer from spectral interference, which occurs in flame emission
spectroscopy.
It is independent of flame temperature.
By atomic absorption technique, traces of one element can easily be determined
in presence of high concentration of other elements.
It has proved very successful in the analysis of bronze and copper alloys and in
the determination of metals like platinum, gold etc.

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 POTENTIOMETRY
For the reaction;

A method of determination of the concentration of a solution by measuring EMF using potentiometer is called Potentiometry.

Principle: Redox titrations can be carried out potentiometrically using platinum and calomel electrode
combination in a manner similar to acid-base neutralizations.

For the reaction; Reduced form  Oxidised form  n electrons

The potential is given by Nernst equation

0.0591 [Oxidised form]
E  Eo  log
n [Reduced form]
0.0591 [Fe3]
E  Eo  log
n [Fe2]

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 Instrumentation
Platinum electrode: An indicator electrode, potential of which changes with change
in concentration metal ions of the solution.
Calomel electrode: a reference electrode and potential of which is fixed.
Beaker: Test solution.
Potentiometer Burette Burette: Standard K2Cr2O7 solution.

Saturated
calomel
electrode
Indicator Procedure: Pipette out 25.0 cm3 of ferrous ammonium sulphate solution into a clean
Electrode
beaker. Add a test tube of dilute sulphuric acid. Immerse the platinum-calomel
Magnetic stirrer
electrode assembly into the beaker containing the test solution. Connect the
electrodes to the potentiometer. Note down the emf of the cell before the addition of
K2Cr2O7. Now add 0.5 ml of K2Cr2O7 from the semi micro burette. Allow the solution
to mix well and measure the potential. Continue the procedure till a sudden rise in
∆𝐸
emf of the cell is observed. Take about 4-5 more readings. Plot a graph of vs volume
∆𝑉
of K2Cr2O7 and calculate the volume of K2Cr2O7 consumed by FAS from the graph.

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 Vol.of K2Cr2O7 Emf E ∆V ∆E ∆E/∆V
(V) ml (mv) (V2V1) ml (E2-E1) (mv/ml)

Application of potentiometry: The method is used for different types of titrations

like,
1. Acid base titrations Amount of FASper liter  N FAS  Eq.Wt.of FAS (392)
2. Diazotization titrations  ..................g (say 'a ' )
3. Redox titrations 392 g of FAS contains 55.85 g Fe
4. Complexometric titrations 55.85  a
a g of FAS contains  ..................g
5. Precipitation titration 392
Advantages of Potentiometric titration
1. The method is inexpensive.
2. This method provides accurate results with minimal chances of errors.
3. Can be used for colored, turbid or fluorescent analyte solution.
4. Can be used in the titration of mixture of acids or mixture of bases.

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 Conductometric Titration
It is an electrochemical method of analysis used for the determination or
measurement of the electrical conductance of an electrolyte solution by means of a
conductometer.

Conductometry means measuring the conductivity of an ionic solution

Conductance is ease with which current flows through the solution. It is reciprocal of resistance.
C=1/R Unit is Ω-1 or mho or Siemens

The conductance of the solution is explained by considering ohm’s law.

According to ohm’s law the current flowing through the conductor is directly proportional to voltage an
inversely proportional to the resistance. I = V
R
The resistance of the any conductor is directly proportional to the length and inversely proportional to the
area of cross section of the conductor. R α l/a
R = S (l/a) where S is specific resistance
Therefore 1/S = 1/R (l/a); K = C (l/a), K = specific conductance, Specific conductance is also called
conductivity. It is the conductance of 1cm3 of solution.
It is defined as the conductance of the solution which is place between two electrodes of area 1cm 2 and 1cm
apart.
The conductance of the solution depends on mobility of ion, number of ion, charge of ion, size of ion

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Types of conductance:
Specific conductance (K) is conductance of the solution which are placed
between two electrodes of area 1cm2 and at 1cm apart
κ = 1/R (l/a) κ = Siemen m-1
Equivalence conductance (λ) is the conductance of the solution when 1g
equivalent weight of solution is placed between two electrodes of area 1cm2 at
1cm apart.
Molar conductance (µ) is the conductance of the solution when 1g molecular
weight of solute is placed between two electrodes of area 1cm2 at 1cm apart

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 Electrical conductivity of an electrolyte solution depends on

1) Type of ions (anion and cation)

2) Concentration of ions
3) Temperature
4) Mobility of ions

Principle: Conductomertic titrations are based

on the measurement of conductance of solution,
which is mainly depends on number of ions,
charge on the ions and mobility of the ions.
During the titration fast moving ions are
replaced slow moving ions and alters the
conductance of a solution. Conductance values
upon addition of titrant vs volume of titrant is
plotted and equivalence point is obtained from
the graph.
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 Applications of conductometric titration:
 
a. Strong acid v/s strong base e.g. HCl v/s NaOH

HCl + NaOH 
 NaCl + H2O
If the strong acid like HCl is titrated against a strong
base such as NaOH, the conductance first decreases

Conductivity
due to replacement of highly mobile H+ ions by less
mobile Na+ ions. After the equivalence point, Equivalence point
of HCl or end point
conductivity rapidly rises with further addition of
NaOH because of continuous addition of highly
mobile OH- ions. A plot of conductance against the
volume of base added is shown in the figure. The
Volume of NaOH solution
point of intersection of two curves gives the end
point.

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 b. Strong acid v/s weak base e.g. HCl v/s NH4OH

HCl + NH4OH 
 NH4Cl + H2O

Conductivity of the solution decreases due to the

replacement of highly mobile H+ ions by NH4+ ions
but after equivalence point, conductivity almost
remains constant, since NH4OH is weak electrolyte
and ionizes to small extent giving very small
conductivity. A plot of conductance against the
volume of base added is shown in the figure. The
point of intersection of two curves gives the
endpoint.

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 c. Weak acid v/s Strong base Eg: CH3COOH against NaOH.

CH3COOH+ NaOH 
 CH3COONa + H2O

The conductance of the acid will be initially low since

acetic acid is a weak electrolyte. When NaOH is added
to the acid, the salt formed is highly ionized and the

Conductivity
conductance increases. On complete neutralization of the
acid, further addition of base leads to increases in the
Equivalence point
number of mobile OH ions. Hence the conductance
-
or end point
increases sharply. A plot of conductance against the
Volume of NaOH solution
volume of base added is shown in the figure. The point
of intersection of two curves gives the endpoint.

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 d. Mixture of strong acid and weak acid v/s strong base HCl, CH3COOH v/s NaOH

HCl + NaOH 
 NaCl + H2O
CH3COOH+ NaOH 
 CH3COONa + H2O
HCl is titrated against a strong base such as NaOH, the conductance first
decreases due to replacement of fast moving H+ ions by slow moving Na+
ions The weak acid does not get neutralized initially because of the well-
known common ion effect.CH3COOH ionizes gradually after the first end
point and a slight increase in the conductance due to salt formation. After
the neutralization point of HCl, CH3COOH, conductivity rapidly rises Equivalence point of CH3COOH
with further addition of NaOH because of continuous addition of fast
moving OH- ions. A plot of conductance against the volume of base Equivalence point
added is shown in the figure. The point of intersection of two curves of HCl
Volume of NaOH solution
gives the neutralization point or endpoint.

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 Advantages:
 Mixture of acid can be titrated
 Indicators are not used
 Very weak acids can be titrated
 Can be used with colored solution.
 

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 THANK YOU

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