solution and determine the volume of copper sulphate

Module V: Instrumental methods of
analysis: Theory, Instrumentation and
applications of Colorimetry, Potentiometry,
Conductometry (Strong acid with a strong base,
weak acid with a strong base, mixture of strong
acid and a weak acid with a strong base), Flame
Photometry, Atomic Absorption Spectroscopy.
1. Theory, Instrumentation and applications of
Colorimetry
Theory: When a monochromatic light of intensity I0 is
incident on a transparent medium, a part, Ia of it is
absorbed, a part, Ir is reflected and the remaining part, It
is transmitted.
I0 = Ia + Ir + It
For a glass - air interface I r is negligible, therefore,
I0 = Ia + It
It / I0 = T called the transmittance, log I/T = log I0 / It is
called the absorbance or optical density.
Colorimetry measurements are based on Beer-Lambert’s
law. This law gives the relation between absorbance A,
concentration c (expressed in mo1/dm3) and path length t,
(ex-pressed in cm).
Beer-Lambert’s law: When a monochromatic light
passes through a transparent medium, the amount of light
absorbed is directly proportional to the concentration and
path length of the solution.
A ∝ Cl
A= logI0/It = εct
Where ε is the molar extinction coefficient, c is the
concentration, t is the path length and is a constant for a
given substance at a given wavelength. If t, the length is
kept constant, then, A∝ c. Hence a plot of absorbance
against concentration gives a straight line.
Instrumentation: Photoelectric colorimeter consists of
(i) Tungsten lamp as the light source.
(ii) A filter which provides the desired wavelength range
wherein the solution gives the maximum absorbance.
(iii) A sample cell
(iv) A photocell detector
solution in the test sample as shown in the figure and find
the amount of copper present in it.
Advantage
(i) Colorimetry gives accurate results at low
concentration. (ii) Colorimetry is also applied for
biological samples.
Theory, Instrumentation and applications of
Potentiometry
Theory : The estimation of concentration of substances
in solution by the measurement of emf is known as
potentiometric titration.
When a metal M is immersed in a solution containing its
own ions Mn+, the electrode potential is given by Nernst
equation.
Thus, the concentration can be calculated, provided Eo of
the electrode is known. If an electrode of the metal
reversible with respect to the corresponding ions is placed
in the solution, the potential will vary throughout the
titration. Initially the change in potential will be small. At
the equivalence point, there will be a steep rise in the
potential. The equivalence point can be determined by
plotting the change in potential against volume of titrant
added.
Instrumentation: A potentiometer consists of: (i)
Calomel electrode as a reference electrode, (ii) Platinum
electrode as an indicator electrode, (iii) a device for
measuring the potential and (iv) magnetic stirrer.

Fig: Schematic layout of colorimeter

Application: Colorimetry is versatile method of
determining the concentration of metals and nonmetals
present in small quantities in ores, soil, samples and
alloys. Eg. Colorimetric estimation of cu in CuSO4. Draw
out 2, 4, 6, 8, and 10 ml cm3 of the Copper sulphate
solution into 50cm3 volumetric flask. Add 5cm3 of
ammonia solution to each of them and dilute upto the
mark with distilled water and mix well. Prepare a blank
solution by diluting 5cm3 of ammonia solution in 50cm3
volumetric flasks. For test solution add 5ml of NH3 and
Application: Potentiometric estimation of FAS using
standard K2Cr2O7 solution. Pipette out 25ml of FAS into
make up to the mark. Measure the absorbance of each of a beaker. Add 1 t.t dil H 2SO4, immerse calomel electrode
these against blank solution at 620 nm. Plot a graph of + platinum electrode into it. Connect the assembly to a
absorbance (OD) against volume of copper sulphate potentiometer and measure the potential by adding
K2Cr2O7 in the increments of 0.5ml.
Plot graph E/ V against volume of K2Cr2O7, and
determine the equivalence point. From the normality and
volume K2Cr2O7, solutions calculate the normality and
the weight of FAS in the given solution.
Advantages of potentiometric titrations are:
(a) They give results more reliable than those obtained
from titrations using indicators
(b) The method is applicable to both coloured and turbid
solutions also
Application: Pipette out 50ml of sample (Acid
mixture/Strong Acid/Weak Acid) into a beaker. Immerse
the conductivity cell into it. Connect the conductivity cell
to a conductivity meter and measure the conductance by
adding NaOH from the burette by increment of 1 ml. Plot
a graph of conductance against volume of NaOH.
Determine the neutralization point from the graph as
shown below in three cases
Case i) Mixture of acids with a Strong base
[(HCl+CH3COOH) Vs NaOH]: The conductance
decreases upon adding NaOH to acid mixture due to
substitution of highly mobile H+ ions by the less mobile
Na+ ions. This trend continues till all the H+ ions of HCl
replaced i.e, the strong acid is neutralized. Continued
addition of NaOH raises the conductance moderately, as
the weak acid (CH3COOH) is converted into its salt
(CH3COONa). Further addition of NaOH raises the
conductance steeply due to the presence of OH-ions. The
titration curve in the graph given determines the location
of the equivalence points.

Theory, Instrumentation and applications of

Conductometry
Theory: Conductometry is based on Ohm’s law which
states that the current i (amperes) flowing in a conductor
is directly proportional to the applied electromotive force,
E (volts), and inversely proportional to the resistance R
(ohms) of the conductor.
E
i
R
The reciprocal of the resistance is called the conductance.
Specific conductance of a solution is defined as the
conductance of a solution present between two parallel ii) Strong acid with a Strong base (HCl Vs NaOH): In
electrodes which have 1cm2 area of cross section and case of a strong acid and a strong base, the conductance
which have kept 1 cm apart. first falls, due to the replacement of highly mobile H+ ion
The principle underlying conductometric titrations is the by the added cation. After the neutralization point, the
substitution of ions with a specific mobility by ions of conductance rapidly rises with further additions of strong
another specific mobility. Therefore, the conductance of base and is due to increase in the concentration of the
solution depends on the number of mobility of ions. The OH- ions.
equivalence point is determined graphically by plotting
conductance against titer values.

Instrumentation: Conductometer consists of

conductivity cell having two platinum electrodes and a
conductance measuring device. The two electrodes have
unit area of cross section and are placed unit distance
apart. A simple arrangement of conductometric titration
is depicted in figure. The solution to be titrated is taken in
the beaker.
iii) Weak acid with a strong base: (CH 3COOH Vs
NaOH): In the conductometric titration of a weak acid
with a strong base, the conductance of the acid will be
initially low due to poor dissociation of acetic acid. On
complete neutralization of the acid, further addition of
base leads to an increase in the number of more mobile
OH- ions. Hence conductance increases sharply.
Advantages:
i) Mixture of acids can be titrated more accurately by
conductometric titration.
ii) Conductometric titrations may be applied where
potentiometric methods fail.
iii) Accurate in dilute solution as well as in more
concentrated solution.
iv) It can be employed with colored solutions.
v) Very weak acids which cannot be titrated
potentiometrically in aqueous solutions can be titrated
conductometrically with relative ease.
Theory, Instrumentation and applications of Flame
Photometry
Principle: Flame photometry is an atomic emission
technique used for detection of metals. If a solution
containing metallic salts is aspirated into a flame, a
vapour, which contains metallic atoms, will be formed.
The electrons from the metallic atoms are then excited
from ground state (E1) to higher energy state (En) where
n=2, 3, 4,…..7, by making use of thermal energy of
flame. From higher energy states these electrons will
return to the ground state by emitting radiations (En – E1=
hν where n=2,3,4……7), which are the characteristic of
each element.
NaCl (s) NaCl (g) Na (g) + Cl (g)
Dissociation energy
Na (g) Na (g) [excited] Na (g)
Excitation energy Emission (hν)
Flame photometer correlates the emitted radiations with
the concentration of these elements. It is simple and rapid
method for the elements that can be easily excited
(Sodium and other alkali metals)
A flame photometer is composed of the pressure regulator
and flow meter for fuel gases an atomizer, burner, optical
system, photosensitive detector and out put recorder. A
filter of the element whose concentration is to be
determined is inserted between the flame and the
detector. Propane gas is used as fuel and air or oxygen is
used as oxidant. Combination of these two will give a
temperature of 19000C. The whole analysis depends on
the flow rate of the fuel, oxidant, the rate of introduction
of the sample and droplet size.
The sample containing the analyte is aspirated into the
flame through atomizer. Radiation from resulting flame is
collected by the lens and allowed to pass through an
optical filter, which permits only the radiation
characteristic of the element under investigation into the
photocell. The output from the photocell represents the
concentration and nature of the element.
Theory – Emission of characteristic radiation by an
element and the correlation of the emission intensity with
concentration of the element form the basis of flame
photometry.
When a solution containing sample element or ion
is aspirated into the flame, following changes takes place,
a) Firstly, solvent gets evaporated leaving behind
salt in the flame.
b) Then, salt gets evaporated into salt vapours,
which further undergo dissociation into its
constituent atoms.
c) Some of the gaseous atoms formed may absorb
heat energy from flame and get electronically
excited to their higher energy level. Being
unstable in the excited state, atoms fall back to
their ground state, in form of light radiation.
d) Intensity of emitted light is proportional to
number of atoms in the excited state, which in
turn is proportional to the concentration of
solution fed into the flame.
e) Different metals emit their characteristic
radiations at different wavelengths, they do not
interfere with each other, even when they are
present together.
Series of changes taking place at the flame are
summarized as follows:
Evaporation
Vaporization Dissociation
M+X- → M+X- → MX →
MX → M (gas) + X (gas)
Solution Mist Solid
Gas
Intensity of emitted radiation, measured as detector
response is related to the concentration by an
expression similar to Beer’s relation,
E=kc
E = detector response
k = constant
 = efficiency of atomic excitation
c = concentration
Instrument: Flame photometer consists of an atomizer,
mixing chamber, burner, filter, detector and a display
device. Pressurized air is passed into atomizer and due to
suction sample solution is drawn into the atomizer. Inside
atomizer it mixes with air stream as a fine mist and passes
into the mixing chamber. In mixing chamber it mixes
with gas and then passes into burner where mixture is
burnt. The emitted radiation from flame passes through
lens and then through a filter which allows only radiation
characteristic of element under study to pass through
detector. The output from detector is read out on a display
device.
Procedure:
Quantitative analysis by flame photometry is done by
calibration curve method. For this, a series of standard
solution of analyte is prepared, aspirated into flame and
emission of each solution is measured in flame
photometer. Then calibration curve is obtained by
plotting emission intensity against concentration of
standard solutions. Sample solution is also properly against concentrations or volume of the solution to form
diluted and then its emission is measured. From the calibration curve. Using the curve obtained find out
calibration curve, concentration of sample solution can be the volume of the unknown solution containing sodium
determined. ions .and calculate the amount of sodium ions in it.

Procedure for determination of potassium Preparation

of standard solutions for calibration
curve: Dissolve exactly 1.090gm of
potassium chloride in water and make
up to 1 liter. This contains 1mg per ml
(1000 ppm). Transfer 2, 4, 6, 8 and
10cm3 of standard potassium solution
into different 25-cm3 volumetric flasks
from a burette. Make up all the
solutions using distilled water. Stopper
the flasks and shake well to get
uniform concentration. To the given
unknown solution also add distilled
water and shake well. Switch on the
instrument, turn the gas supply on and
light the gas at the burner. Adjust the
air supply from the compressor to 10
lbs/sq inch using pressure regulator knob. Place the
Application: potassium filter in position. Now dip the capillary tube in
1) Flame photometer can be applied both for a cell containing distilled water. The stream of air
quantitative and qualitative analysis of atomized as a fine mist draws up the liquid. Regulate the
elements. The radiations emitted by the flame gas supply so that the colour of the flame completely
photometer are characteristic to particular turns to blue. Adjust the flame photometer to zero by
metal. Hence with the help of Flame means of zero control knob. Now feed the 100 ppm
photometer we can detect the presence of any potassium solution and adjust the reading to hundred.
specific element in the given sample. Repeat the process to confirm the accuracy of the
2) The presence of some group II elements is calibration.
critical for soil health. We can determine the
presence of various alkali and alkaline earth Feed the various potassium solutions prepared through
metals in soil sample by conducting flame test the flame by spraying with atomizer one by one including
and then the soil can be supplied with specific
fertiliser.
Advantage:
1) The method of analysis is very simple and
economical.
2) It is quick, convenient, selective and sensitive
analysis.
3) It is both and qualitative and quantitative in
nature.
4) Even very low concentrations (parts per
million/ppm to parts per billion/ppb range) of
metals in the sample can be determined.
Procedure for determination of sodium: Transfer 2, 4,
6, 8 and 10cm3 of standard sodium solution into different
25-cm3 volumetric flasks from a burette. Make up all the
solutions using distilled water. Stopper the flasks and
shake well to get uniform concentration. To the given
unknown solution also add distilled water and shake well. the unknown solution. Note down the flame photometer
Switch on the instrument, turn the gas supply on and light reading. Plot a graph of flame photometer readings
the gas at the burner. Adjust the air supply from the against concentrations or volume of the solution to form
compressor to 10 lbs/sq inch using pressure regulator the calibration curve. Using the curve obtained find out
knob. Place the sodium filter (589nm) in position. Now the volume of the unknown solution containing potassium
dip the capillary tube in a cell containing distilled water. ions and calculate the amount of potassium ions in it.
The stream of air automised as a fine mist draws up the
liquid. Regulate the gas supply so that the colour of the
flame completely turns to blue. Adjust the flame
photometer to zero by means of zero control knob. Now
feed the 100 ppm sodium solution and adjust the reading
to hundred. Repeat the process to confirm the accuracy of
the calibration.
Feed the various sodium solutions prepared through the
flame by spraying with automiser one by one including
the unknown solution. Note down the flame photometer
reading. Plot a graph of flame photometer readings