Wachemo University

College of NCS
Department of Chemistry
Year II – 2nd Semester

Instrumental Analysis I
Chem 2033
 UNIT-1
Analytical Separation Techniques and Classical
Method of Analysis

 Analytical Chemistry deals with methods for determining

the chemical composition of samples.

• Qualitative Analysis- provides information about the

identity of species or functional groups in the sample.
• Quantitative Analysis- provides numerical information of
analyte.
2
 Analytical methods can be separated into classical and
instrumental. Classical methods use separations such as precipitation,
extraction, and distillation and qualitative analysis by color, odor, or
melting point.
• Classical Methods: Wet chemical methods such as precipitation, extraction,
distillation, boiling or melting points, gravimetric and titrimetric measurements.
• Instrumental Methods: Analytical measurements (conductivity, electrode
potential, light absorption or emission, mass-to-charge ratio, fluorescence etc.) are
made using instrumentation.
 The majority of the classical analytical methods rely on chemical reactions
to perform an analysis. In contrast, instrumental methods typically depend
on the measurement of a physical property of the analyte.

 Types of Instrumental Methods

1. Spectroscopic methods:

a. Atomic spectroscopy

b. Molecular spectroscopy

2. Chromatographic methods (separations)

3. Electroanalytical chemistry:
 include potentiometry, amperometry, conductometry,
electrogravimetry, voltammetry (and polarography), and coulometry. It
3
reflect the measured electric property or its units.
 UNIT-2
INTRODUCTION TO CHROMATOGRAPHIC
SEPARATION

 In 1906 Tswett used chromatography to separate plant

pigments

 He called this new technique ‗chromatography‘ because

the result of the analysis was 'written in color' along the
length of the adsorbent column

 Chroma means ―color and graphein means to ―write

5
 Introduction to Chromatography

☞Chromatography is a technique in which the components of a

mixture are separated based on differences in the rates at
which they are carried through a stationary phase by a mobile
phase.

☞ Components:
• Mobile phase
• Stationary phase
• Supporting medium

6
 Introduction to Chromatography…Cont’d

• The stationary phase- is a phase that is fixed in place either in

a column or on a planar surface.
• The mobile phase- is a phase that moves over or through the
stationary phase carrying with it the analyte
mixture.
☞ The mobile phase may be a gas, a liquid, or a
supercritical fluid.
• Supporting Medium- is a solid surface on which the stationary
6
phase is bound or coated.
 Classification of Chromatographic Methods
• Based on the type of supporting medium
chromatographic methods are of two basic types
1) Column chromatography: the stationary phase is held in a
narrow tube, and the mobile phase is forced through the tube
under pressure or by gravity.
2) Planar chromatography: the stationary phase is supported on
flat plate or in the pores of a paper, and the mobile phase
moves through the stationary phase by capillary action or
7
under the influence of gravity
 Types of Chromatographic Methods…Cont’d
• Chromatographic methods fall into three categories
based on the nature of the mobile phase: liquid, gas,
and supercritical fluid.
• There are five types of liquid chromatography and two
types of gas chromatography that differ in the nature of
the stationary phase and the types of equilibria between

phases.

9
Classification of Column Chromatographic Methods

1
0
Planar chromatography
• Planar chromatographic methods include
☞Paper chromatography (PC)
☞Thin-layer chromatography (TLC)

 Paper chromatography (PC)

• is a liquid-liquid chromatography
• is a partition chromatography
• Substances to be identified are ‗spotted‘ near one end of the
filter paper
• As the solvent moves up the paper, different molecules move
at different rates 10
• Sample is spotted at the bottom • Solvent migrates in the paper

line of the paper and elutes the solutes

• Paper is placed in a tank filled • The solute migrate depending

with 1 cm solvent on their affinity for the solvent
11
Mark the solvent
front & allow
paper to dry Solvent
front

Purple spots
develop located at
different distances
from the origin line

origin12
Spots
solvent
☞When the spots are colourless, a locating agent (like ninhydrin)
is needed to visualise their positions on the chromatography
paper
☞The chromatogram can be analysed by measuring the distance
travelled by the solvent front, and the distance from the origin to
the centre of each spot.
• This is used to calculate the Rf (retardation factor) value for each spot:
𝒅𝒊𝒔𝒕𝒂𝒏𝒄𝒆 𝒎𝒐𝒗𝒆𝒅 𝒃𝒚 𝒔𝒐𝒍𝒖𝒕𝒆
𝑹𝒇 =
𝒅𝒊𝒔𝒕𝒂𝒏𝒄𝒆 𝒎𝒐𝒗𝒆𝒅 𝒃𝒚 𝒔𝒐𝒍𝒗𝒆𝒏𝒕 𝒇𝒓𝒐𝒏𝒕

✈ An Rf value is characteristic of a particular solute in a particular solvent.

✈ It can be used to identify components of a mixture by comparing to tables
13
of known Rf values.
✈The Rf value is always a value less than one as the solvent
front always moves further than the solute molecules

Example:

14
  Thin Layer chromatography (TLC)

 Thin Layer chromatography:

• The modern version of paper chromatography
• Paper is replaced by a layer (100-200 μm) of stationary
phase (silica gel, alumina) coated on a rectangular glass
plate (10-20 cm large)
• Can be considered a form of liquid-solid chromatography
=> adsorption chromatography
 Principles of Thin-Layer chromatography:
• Typical thin-layer separations are performed on a glass plate that
is coated with a stationary phase, which consists of a thin and 15
adherent layer of finely divided particles.
17
 Cont’d

 Sample Application
• Sample application is the most critical aspect of TLC.
• Usually the sample is applied as a spot 1 to 2 cm from the edge
of the plate.
• Manual application of samples is performed by touching a
capillary tube containing the sample to the plate or by use of a
syringe.
• Mechanical dispensers, which increase the precision and
accuracy of sample application, are available commercially.
18
 Cont’d
• Plate development is the process by which a sample is carried
through the stationary phase by a mobile phase.

• After applying a spot, the plate is placed in a closed

container saturated with vapors of the developing solvent.

• One end of the plate is immersed in the developing solvent,

with care being taken to avoid direct contact between the sample
and the developer.

• After the developer has traversed one half or two thirds of the
18
length of the plate, it is removed from the container and dried.
 Cont’d

 Locating Analytes on the Plate

• Several methods are used to locate sample components after
separation.
• Two common methods that can be applied to most organic
mixtures involve spraying with chemicals or using UV.

• These reagents react with compounds to yield colored products.

• Reagents such as ninhydrin, iodine, sulfuric acid etc. are useful
for locating separated species.

☞The process of locating analytes on a thin-layer plate is 19

often termed visualization.

 In order to use TLC as a quantitative method of analysis, it is
essential to quantify the spots, along with definitions for all of
the usual parameters:

 specificity
 linearity
 precision etc.
 This is done by placing the plate under the lens of a
densitometer that can measure either absorption or
fluorescence at one or several wavelengths.
 This instrument produces a pseudo-chromatogram that
20
contains peaks whose areas can be measured.
 Efficiency of separation
 Efficiency is related experimentally to a solute‘s peak width.
 an efficient system will produce narrow peaks smaller
difference in interactions in order to separate two solutes
 Efficiency is related theoretically to the various kinetic
processes that are involved in solute retention and transport in
the column

 determine the width or standard deviation (σ) of peaks

o Estimate s from peak widths,
Wh assuming Gaussian shaped peak:
Wb = 4σ

Wh = 2.354σ
21

 Dependent on the amount of time that a solute spends in the column

Number of theoretical plates (N): compare efficiencies of a
system for solutes that have different retention times
N = (tR/σ)2
or for a Gaussian shaped peak

N = 16 (tR/Wb)2

N = 5.54 (tR/Wh)2
 The larger the value of N:
 the better the column will be able to separate two
compounds.
 the better the ability to resolve solutes that have small
differences in retention 22
 N is independent of solute retention but, depends on the length of
the column

Plate height or height equivalent of a theoretical plate (H or

HETP): compare efficiencies of columns with different lengths:
H = L/N
where: L = column length
N = number of theoretical plates for the column
Note: H simply gives the length of the column that corresponds to
one theoretical plate 23
 H can be used to relate various chromatographic parameters to
the kinetic processes that give rise to peak broadening due to
the following reasons:

a. Eddy diffusion d. Stationary phase mass transfer

b. Mobile phase mass transfer e. Longitudinal diffusion
c. Stagnant mobile phase mass
transfer

25
a) Eddy diffusion – due to the presence of multiple flow paths
through a packed column.
As solute molecules travel through
the column, some arrive at the end
sooner then others simply due to the
different path traveled around the
support particles in the column that
result in different travel distances.
Longer path arrives at end

26
b) Mobile phase mass transfer – caused by the presence of
multiple flow between particles of the support in the column.

 A solute in the center of the

channel moves more quickly than
solute at the edges, it will tend to
reach the end of the channel first
leading to band-broadening

 The degree of band-broadening due to eddy diffusion and

mobile phase mass transfer depends mainly on:

1) the size of the packing material

2) the diffusion rate of the solute 26
c) Stagnant mobile phase mass transfer – due to differences in the
rate of diffusion of the solute molecules between flowing mobile phase
stagnant mobile phase.

 Since a solute does not travel down the

column when it is in the stagnant mobile
phase, it spends a longer time in the column
than solute that remains in the flowing mobile
phase.
 The degree of band-broadening due to this depends on:
1) the size, shape and pore structure of the packing material
2) the diffusion and retention of the solute 27

3) the flow-rate of the solute through the column

d) Stationary phase mass transfer – due to the movement of solute
between the stagnant phase and the stationary phase.

 Since different solute molecules spend

different lengths of time in the
stationary phase, they also spend
different amounts of time on the
column, giving rise to band-
broadening.
 The degree of band-broadening due to this depends on:
1) the retention and diffusion of the solute
2) the flow-rate of the solute through the column 28

3) the kinetics of interaction between the solute and stationary phase

 e) Longitudinal diffusion – due to the diffusion of the solute along
the length of the column in the flowing mobile phase.

 The degree of band-broadening due to longitudinal diffusion depends on:

1) the diffusion of the solute
2) the flow-rate of the solute through the column
30
Van Deemter equation: relates flow-rate or linear velocity to H:
H = A + B/µ + Cµ
where:
µ = linear velocity (flow-rate x Vm/L)
H = total plate height of the column
A = constant representing eddy diffusion & mobile phase mass transfer
B = constant representing longitudinal diffusion
C = constant representing stagnant mobile phase & stationary phase
mass transfer

31
 One use of plate height (H) is to relate these kinetic process to
band broadening to a parameter of the chromatographic system
(e.g., flow-rate).
 This relationship is used to predict what the resulting effect
would be of varying this parameter on the overall efficiency of
the chromatographic system.

Number of theoretical plates(N) (N) = 5.54 (tR/Wh)2

H = L/N

32
Chromatography is used by scientists to:

•Analyze – examine a mixture, its components, and their

relations to one another

•Identify – determine the identity of a mixture or components

based on known components

• Purify – separate components in order to isolate one of

interest for further study

•Quantify – determine the amount of the a mixture and/or the

components present in the sample
33
 Applications of chromatography:
• Pharmaceutical Company - determine amount of each chemical
found in new product
•Hospital - detect blood or alcohol levels in a patient‘s blood
stream
•Law Enforcement – to compare a sample found at a crime scene
to samples from suspects
•Environmental Agency – determine the level of pollutants in
the water supply
•Manufacturing Plant – to purify a chemical needed to make a
product

34
 UNIT-3
GAS CHROMATOGRAPHY (GC)
Principles of GC
 In GC, the sample is vaporized and injected onto the head of a
chromatographic column.
 Elution is brought about by the flow of an inert gaseous mobile
phase.
 The mobile phase function is to transport the analyte through the
column.

 Gas-liquid chromatography is based upon the partition of the

analyte between a gaseous mobile phase and a liquid phase
34
immobilized on the surface of an inert solid.
 TYPES OF GC
 Two types of GC are encountered: gas-liquid chromatography (GLC) and
gas-solid chromatography (GSC).
 In GLC, the mobile phase is a gas, and the stationary phase is a liquid
that is retained on the surface of an inert solid by adsorption or chemical
bonding.
 GLC finds widespread use in all fields of science where its name is
usually shortened to GC.
 In GSC, the mobile phase is a gas, and the stationary phase is a solid
that retains the analytes by physical adsorption.
 GSC permits the separation and determination of low-molecular-mass
gases, such as air components, hydrogen sulfide, carbon monoxide, and
35
nitrogen oxides.
INSTRUMENTS FOR GC

 A simple GC system consists of:

1. Gas source
2. Injector or sample application system
3. Chromatographic column
4. Detector
5. Computer or recorder

37
Block diagram of a typical Gas Chromatograph.
38
1. Carrier Gas-Supply
• The mobile phase gas in GC is called the carrier gas and
must be chemically inert.

 He is the most common mobile phase, although Ar,

N2, and H2 are also used.

 These gases are available in pressurized tanks.

 Pressure regulators, gauges, and flow meters are
required to control the flow rate of the gas.

 In addition, the carrier gas system often contains a

molecular sieve to remove water or other impurities. 38
2. Sample injection system
 For high column efficiency, a suitable sized sample should be introduced
as a plug of vapor

 Slow injection or oversized sample injection cause band spreading

and poor resolution.
 The most common method of sample injection involves the use of micro
syringe to inject a liquid or gaseous samples through a silicon-rubber
diaphragm or septum into a heated sample port located at the head of the
column.

 The sample port is usually kept at about 50ºC greater than the boiling point
of the least volatile component of the sample.

40
 C O N T ’D

Sample Injection Port

41
3. Column Configurations and Column Ovens
 The columns in G C are of two general types: packed
columns or capillary columns.

42
 C O N T ’D
• Chromatographic columns vary in length from less than 2 m
to 60 m or more.

• Columns are constructed of stainless steel, glass, fused silica,

or Teflon.
• In order to fit into an oven for thermostating, they are usually
formed as coils having diameters of 10 to 30 cm.

Column Ovens
Column temperature is an important variable that must be
controlled to a few tenths of a degree for precise work.
43
 Thus, the column is ordinarily housed in a thermostated oven.
 The optimum column temperature depends upon the boiling
point of the sample and the degree of separation required.
 Roughly, a temperature equal to or slightly above the average
boiling point of a sample is recommended for (2 to 30 min).
 For samples with a broad boiling range, it is often desirable to
employ temperature programming, whereby the column
temperature is increased either continuously or in steps as the
separation proceeds.

44
4. Detection Systems
The ideal detector for GC has the following characteristics:
1. Adequate sensitivity
2. Good stability and reproducibility.
3. A linear response to solutes that extends over several orders of
magnitude.

4. A temperature range from room temperature to at least 400oC.

5. A short response time that is independent of flow rate.
6. High reliability and ease of use.
7. A highly selective response
8. Nondestructive of sample. 44
 Gas Chromatographic Detectors
Type Applicable Samples
Flame ionization Hydrocarbons
Thermal conductivity Universal detector
Electron capture Halogenated compounds
Mass spectrometer (MS) Tunable for any species
Thermionic Nitrogen and phosphorous
compounds
Electrolytic conductivity Compounds containing
halogens, sulfur, or nitrogen
Photoionization Compounds ionized by UV
radiation
Fourier transform IR (FTIR) Organic compounds 46
 Qualitative Analysis

• Gas chromatograms are widely used as criteria of purity for

organic compounds.
• Contaminants, if present, are revealed by the appearance of
additional peaks; the areas under these peaks provide rough
estimates of the extent of contamination.

• Retention times should be useful for the identification of

components in mixtures.
• GC provides an excellent means of confirming the presence or
absence of a suspected compound in a mixture.
47
 Quantitative Analysis
• Quantitative GC is based on comparison of either the height or
the area of an analyte peak with that of one or more standards.

☞ If conditions are properly controlled, both of these

parameters vary linearly with concentration.
• Most modern chromatographic instruments are equipped with
computers that provide measurements of relative peak areas.

☞ If such equipment is not available, a manual estimate must

be made.

48
 CONT’D
 Calibration with Standards
• In the most straightforward method for quantitative GC
analyses, a series of standard solutions that estimate the
composition of the unknown should be prepared.
• Chromatograms for the standards are then obtained, and peak
heights or areas are plotted as a function of concentration to
obtain a working curve.
• A plot of the data should yield a straight line passing through
the origin and quantitative analyses are based on this plot.
• Frequent standardization is necessary for the highest accuracy.
49
 UNIT-4
LIQUID CHROMATOGRAPHY (LC)
Principle
 Liquid Chromatography (LC) is a chromatographic technique in which
the mobile phase is a liquid.
 Example:
 Paper chromatography,
 Thin Layer Chromatography,
 High Performance Liquid Chromatography(HPLC) etc.

 Retention of solutes in LC depend on their interaction with both the

mobile phase and stationary phase.
 LC is more flexible in optimizing separations  change either stationary 49
or mobile phase
 For column chromatography, solutes can be eluted from a column by
using either a constant column conditions or gradient elution
Isocratic elution: use of a constant mobile phase composition to elute
solutes

Gradient elution: changing the composition of the mobile phase with time
 In choosing a mobile phase for LC, several factors need to be considered
– type of stationary phase used
– solubility of the solutes
– viscosity of the mobile phase
– type of detector used and solvent's background signal
– purity of the solvents
– miscibility of the solvents (for gradient elution)
50
 Instruments for LC

51
 Basic components of versatile LC includes: solvent, pump, injector, column,
detector, and computer.
 Types of Liquid Chromatography (Retention Mechanisms )

 Techniques in LC are classified according to the method of solute separation

i. Adsorption chromatography
iv. Size-exclusion chromatography
ii. Affinity chromatography
v. Ion-exchange chromatography
iii. Partition chromatography

1. Adsorption chromatography (Liquid-solid Chromatography)

 The stationary phase is a bonded rigid silica or silica based component or
alumina onto the inside of the column.

 The analytes that are in the mobile phase having an affinity for the
stationary phase will be adsorbed onto it and those that do not will pass
through having shorter retention times.

 Both normal and reverse phases of this method are applicable. 52

 A LC technique which separates solutes based on their adsorption to solid
particles.

 This technique is suited for non-polar small compounds (MW<5000).

 One advantages of adsorption chromatography, is that it is able to separate
some compounds that can not be separated by other methods.

53
2. Partition chromatography (Liquid-liquid Chromatography)

 Both the stationary phase and the mobile

phase are liquid.
 The stationary phase liquid would be an
immiscible liquid with the mobile phase.

 It is a Chromatographic technique in which

solute are separated based on their partition
Phas e 2 Phase 2
between a liquid mobile phase and a liquid
Phas e 1 Phase 1
stationary phase coated on a solid support.

How can ions be separated by using partition Chromatography?

Normal Phase LC and Reversed-phase LC 54

Stationary phase: polar non-polar

3. Ion-Exchange Chromatography (Ion Chromatography)
 Applied to separate and determine ions on columns that have a low ion
exchange capacity.
 This is based on the equilibria of ion exchange between the ions in solution
and the counter ions to pair with the oppositely charged ions that are fixed to
the stationary phase.
 Solutes are separated by their adsorption onto a support containing fixed
charge onto a support containing fixed charges on its surface.

55
4. Affinity chromatography

 Involves binding a reagent to the analyte molecules in a sample.

 After the binding, only the molecules that have this ligand are retained in the
column, the unbound analyte is passed through in the mobile phase.

 The stationary phase is usually agarose or a porous glass bead that is able to
immobilize the bonded molecule.

For biomolecule separation

57
5. Size-exclusion Chromatography for polymer and bio-polymer
 Done by having the stationary phase be packed with small particles of silica
or polymer to form uniform pores.
 The smaller molecules will get trapped in the silica particles and will elude
from the column at a rate that is greater than that of larger molecules.
 Thus, the retention time depends on the size of the molecules.
 Larger molecules will be swept away in the mobile phase, therefore having
a smaller retention time.

 Is a powerful technique that is particularly applicable to high-

molecular-mass species.
✈Separation is based on the size (bulkiness) of molecules.

58
Of two types:
i. Gel filtration- is a type in which the packing is hydrophilic.
- is used to separate polar species.
ii. Gel permeation- is a type in which the packing is hydrophobic.
- is used to separate nonpolar species.

58
 LC Method Development

Problem Definition

Mode selection What type of LC should be used?

Stationary phase and mobile phase,

Selectivity Optimization Temperature

Column length, particle size, flow rate,

System optimization instrument configuration, sample injection…

Method Validation Accuracy, sensitivity, specificity, detection

Limit, quantification limit, linearity 59
 UNIT-5
INTRODUCTION TO ELECTRO-ANALYTICAL
CHEMISTRY

 Electroanalytical chemistry encompasses a group of

quantitative analytical methods that are based upon the
electrical properties of a solution of the analyte.
 They have certain general advantages:
 specific for a particular oxidation state of an element.
 Instrumentation is relatively inexpensive.
 Provide information about activities rather than
60
concentrations of chemical species.
 Electrochemical Cell
1) Basic Set-up:
a) Two electrodes
b) electrolytes solution
c) external connection between electrodes (wire)
d) different solutions connected by a salt bridge.

salt bridge – acts to isolate two halves of electrochemical cell

while allowing migration of ions and current flow.

61
2) Flow of current (charge) in cell:
a) electrons (e-) within wires between two electrodes
b) ions within solution of each ½ cell (anions & cations) and
through salt bridge

c) electrochemical reactions at electrode

electrons

Cl- K+

Zn2+ Cu2+
SO42- SO42-
62
 At Cu electrode: Cu2+ + 2e- ↔ Cu(s)  reduction – gain of e-
net decrease in charge of species

At Zn electrode: Zn(s) ↔ Zn2+ + 2e-  oxidation – loss of e- net

increase in charge of species
3) Net Reaction in Cell – sum of reactions occurring in the two ½
cells
Zn(s) ↔ Zn2+ + 2e-
Cu2+ + 2e- ↔ Cu(s)

Cu2+ + Zn(s) ↔ Zn2+ + Cu(s)

4) Potential of overall cell = measure of the tendency of this reaction to proceed
to equilibrium, potential (Ecell) = 0
 Larger the potential, the further the reaction is from equilibrium 63
and the greater the driving force that exists
 Types of electrochemical Cells
a) Galvanic Cells – reaction occurs naturally
- positive potential (Ecell = +)
- exothermic  produces energy

Galvanic Cell 64
b) Electrolytic Cells – reaction does not occur naturally, requires
external stimulus (energy) to occur

- negative potential (Ecell = -)

- endothermic  requires energy

External battery at
higher power than cell
potential

Electrolytic Cell

c. Chemically Reversible Cell – a cell in which reversing

65

the direction current simply reverses the chemical reaction

5) Electrodes
a) Cathode – electrode where reduction occurs
e-
Anode – electrode where oxidation occurs
b) Examples of cathode ½ reactions:
Cu2+ + 2e- ↔ Cu(s)
Fe3+ + e- ↔ Fe2+
AgCl(s) + e- ↔ Ag(s) + Cl-
- e- supplied by electrical current via electrode
- species (products/reactants) can both be in solution (Fe3+/Fe2+)
solids or coated on electrodes (AgCl(s)/Ag(s) or combination 66
(Cu2+/Cu(s)
c) Examples of anode ½ reactions:
Cu(s) ↔ Cu2+ + 2e-
Fe2+ ↔ Fe3+ + e-

Ag(s) + Cl- ↔ AgCl(s) + e-

- e- is taken up by electrode into electrical circuit

e-

67
d) Liquid junctions – interface between two solutions with
different components or concentrations

Liquid Junction

 Small potentials may develop at junction that affect overall

cell potential 68
e) Representation of Cells: by convention start with anode on left
Phase boundary
Electrode/solution interface

anode cathode
Zn|ZnSO4(a ZN2+ = 0.0100)||CuSO 4(a Cu2+ = 0.0100)|Cu reduction
oxidation
2 liquid junctions
Solution in contact with due to salt bridge Solution in contact with
anode & its concentration cathode & its concentration

70
f) Electrode Potentials
1) for convenience, represent overall reaction in cell as two ½
reactions

i. one at anode & other at cathode

ii. each ½ reaction has certain potential associated with it
iii. by convention, write both ½ reactions as reduction:
Cu2+ + 2e- ↔ Cu(s) (Ecathode)
Zn2+ + 2e- ↔ Zn(s) (-Eanode)
iv. potential of cell is then defined as:
Ecell = Ecathode – Eanode
71
2) Problem – can not measure potential of just one electrode.
i. need to compare to another electrode
ii. determine potential of all ½ cell reactions vs. a common
reference electrode
iii. reference electrode – standard hydrogen electrode (SHE)
2H+ + 2e- ↔ H2(g)
note: potential affected by pH, [H+] and PH2

72
 By convention, ESHE = 0V at [H+] = 1M, PH2 = 1 atm and at all
temperatures

 Potentials of other electrodes are compared to SHE using

electrode in question as cathode and SHE as anode:
Mn+ + ne- ↔ M(s)
Ecell = Ecathode – Eanode
Ecell = Ecathode – ESHE
 By definition:
72
Ecell = Ecathode – 0
Ecell = Ecathode
Standard Electrode Potential (Eo) – measured Ecell when all
species in solution or gas has an activity of 1.00
Activity (a) – proportional to molar concentration

ax = γx[X]
where:
γx - activity coefficient of solute X
[X] - molar concentration of solute X

 If Eo is ―+‖, it indicates that the reaction:

Mn+ + n/2H2(g) ↔ M(s) + nH+
 is favored or spontaneous.
 If Eo is ―
-‖, it indicates that the reaction:
73
 is not favored or non-spontaneous
 As Eo increases  oxidizing ability of ½ cell reaction increases

Reaction at Interface Half-cell Potential (Eo)

Al3+ + 3e-  Al -1.706 V
Zn2+ + 2e-  Zn -0.763 V
Easily reduced, Better Oxidizing Agent

Cr3+ + 3e-  Cr -0.744

Easily oxidized, Better Reducing Agent

Fe2+ + 2e-  Fe -0.409V
Cd2+ + 2e-  Cd -0.401 V
Ni2+ + 2e-  Ni -0.230 V
Pb2+ + 2e-  Pb -0.126 V
2H+ + 2e-  H2 0.00 V
AgCl + e-  Ag + Cl- +0.223 V
Hg2Cl2 + 2e-  2Hg + 2Cl- +0.268 V
Cu2+ + 2e-  Cu +0.340 V
Ag+ + e-  Ag +0.799 V
Au+ + e-  Au +1.680 V

75
 Currents in Electrochemical Cells
a) Ohm’s Law
E = IR
where:
E = potential (V, voltage)
I = current (amps)
R = resistance (ohms)
 R depends on concentration and types of ions in solution

76
b) Mass Transport Resulting From Current in Bulk Solution
- currents in solution are carried by movement of ions
- small ions move faster and carry more current than larger ions
- species reacting at electrode don‘t have to be only species
carrying current
- example:
 If much higher concentration of other ions in bulk solution‚
 analytes will carry current only in region near electrode
surface

77
c) Currents at Electrode Surfaces
i) Faradic
 transfer of e- to/from electrode by redox reactions
 governed by Faraday’s Law
 Amount of current is proportional to amount of species 77

oxidized or reduced
ii) Non-Faradic Current
 due to processes other than redox reactions at electrodes
 example – charging current
 movement of ions = current
- as system approaches equilibrium  get decrease in ion
movement and current

78
 Types of Electroanalytical Methods

I = current, E = potential, R = resistance, G = conductance, Q = quantity of charge,

79
t = time, vol = volume of a standard solution, m = mass of an electrode
 UNIT- 6
POTENTIOMETRY

 Which electrical property is to be measured in

Potentiometry?

Potentiometric methods of analysis are based on

measuring the potential of electrochemical

cells without drawing appreciable current.

80
 6.1 Basic principles of Potentiometry

| | |
Reference electrode 𝒔𝒂𝒍𝒕 𝒃𝒓𝒊𝒅𝒈𝒆 analyte solution 𝒊𝒏𝒅𝒊𝒄𝒂𝒕𝒐𝒓 𝒆𝒍𝒆𝒄𝒕𝒓𝒐𝒅𝒆
𝑬𝒓𝒆𝒇 𝑬𝒋 𝑬𝒊𝒏𝒅

 The potential of the indicator electrode cannot be measured alone.

 By convention (IUPAC convention for electrode potentials), the

reference electrode is always treated as the left-hand electrode in
potentiometric measurements (Figure 6-1).
8
1
 Components of a
potentiometric cell:

i. Reference electrode
ii. Indicator electrode
iii. Salt bridge
iv. Potentiometer

82
Figure 6-1 A cell for potentiometric determinations.
 The potential of the cell (Figure 6-1 ) is given by the equation:

𝑬𝒄𝒆𝒍𝒍 = 𝑬𝒊𝒏𝒅 − 𝑬𝒓𝒆𝒇 + 𝑬𝒋 … … … … … … … . (6.1)

 𝑬𝒊𝒏𝒅 contains the information that we are looking

for— the concentration of the analyte.

To make a potentiometric determination of an analyte, we must

 measure a cell potential (𝑬𝒄𝒆𝒍𝒍),

 correct this potential for reference & junction potentials, &
 compute the analyte concentration from the 𝑬𝒊𝒏𝒅. 8
3
How can we determine the concentration of the analyte?
How can we determine the concentration of the analyte?

 We can determine the concentration of the analyte only

through proper calibration of the electrode system with

solutions of known concentration.

 Determination of the substances by potentiometric technique can

be carried out by two ways:

1. Direct potentiometry or
2. Potentiometric titrations 8
4
6.2 Instrumentation for Measuring Cell Potential

The components of a potentiometric cell:

i. Reference electrode: provides reference potential (𝑬𝒓𝒆𝒇).

ii. Indicator electrode: responds to species of interest by

developing an indicator potential (𝑬𝒊𝒏𝒅).

iii. Salt bridge: prevents the components of the analyte solution

from mixing with those of the reference electrode.
85
 6.2 Instrumentation …cont’d

iv. Potential measuring device (Potentiometer)

A device for measuring an unknown potential difference or

electromotive force by comparing it to a known standard
potential is called potentiometer.

v. Electrical connection: used to connect the electrodes with

the potentiometer.

87
6.3 Types of electrodes

1. Reference electrodes

2. Indicator electrodes
88
 Reference electrodes
 A reference electrode is a half-cell having a known electrode
potential that remains constant or fixed at constant temperature &
completely insensitive to the composition of the analyte solution.

 Its additional ideal requirements or desired characteristics are:

• should be easy to assemble, & should maintain a constant
potential while passing minimal currents.
• reversible i.e. obeys to Nernst Eqn.

 The common reference electrodes are Calomel Reference

Electrodes & Silver/Silver Chloride Reference Electrodes 8
8
 Reference electrodes
A. Calomel Reference Electrodes

 consist of mercury in contact with a solution that is saturated with

mercury(I) chloride (calomel) & that also contains a known
concentration of KCl.

 Calomel half-cells can be represented as follows:

𝑯𝒈|𝑯𝒈𝟐 𝑪𝒍𝟐 𝒔𝒂𝒕′ 𝒅 , 𝑲𝑪𝒍(𝒙 𝑴)||

where x represents the molar concentration of KCl in the solution.

8
9
 Reference electrodes

A. Calomel Reference Electrodes …cont’d

 The electrode potential for Calomel half-cell is determined by the

rxn.

𝑯𝒈𝟐𝑪𝒍𝟐 𝒔 + 𝟐𝒆− ↔ 𝟐𝑯𝒈 𝒍 + 𝟐𝑪𝒍−(𝒂𝒒)

& depends on the chloride concentration.

 Thus, the KCl concentration must be specified

in describing the electrode. 9
0
Figure 6-2 Diagram of a typical commercial 91
saturated calomel electrode.
 Reference electrodes
B. Silver/Silver Chloride Reference Electrodes
 The most widely marketed reference electrode system consists of
a Ag electrode immersed in a solution of KCl that has been
saturated with AgCl:

𝑨𝒈|𝑨𝒈𝑪𝒍 𝒔𝒂𝒕′ 𝒅 , 𝑲𝑪𝒍(𝒙 𝑴)||

 The electrode potential is determined by the half-reaction

𝑨𝒈𝑪𝒍 𝒔 + 𝒆− ↔ 𝑨𝒈 𝒔 + 𝑪𝒍−(𝒂𝒒)

 Ag–AgCl electrodes have the advantage that they can be used at

9
temperatures greater than 60°C, while calomel electrodes cannot.
2
Figure 6-3 Diagram of a
Ag/AgCl electrode showing the
parts of the electrode that produce
the reference electrode potential,

𝑬𝒓𝒆𝒇, & the junction potential, 𝑬𝒋.

93
 Indicator electrodes
 The indicator electrode, which is immersed in a solution of the
analyte, develops a potential, 𝑬𝒊𝒏𝒅, that depends on the activity of
the analyte.

 An ideal indicator electrode responds rapidly & reproducibly to

changes in the concentration of an analyte ion (or group of analyte
ions).

 Indicator electrodes are remarkably selective in their responses.

 Indicator electrodes are of three types: metallic, membrane, &

ion-sensitive field effect transistors. 9
4
 1. Metallic Indicator Electrodes
 It is convenient to classify metallic indicator electrodes as
electrodes of the first kind, electrodes of the second kind, &
inert redox electrodes.

i. Electrodes of the First Kind

 An electrode of the first kind is a pure metal electrode that

is in direct equilibrium with its cation in the solution.

9
 A single reaction is involved.
5
 i. Electrodes of the First Kind ….cont’d

 For example, the equilibrium between a copper & its cation 𝐶𝑢2+is
𝑪𝒖𝟐+ 𝒂𝒒 + 𝟐𝒆− ↔ 𝑪𝒖 𝒔
for which

𝟎. 𝟎𝟓𝟗𝟐 𝟏
𝑬𝒊𝒏𝒅 = 𝑬𝒐 𝑪𝒖 − 𝒍𝒐𝒈
𝟐 𝒂𝑪𝒖𝟐+
𝟎.𝟎𝟓𝟗𝟐
= 𝑬𝒐𝑪𝒖 + 𝟎.𝟎𝟓𝟗𝟐 𝒍𝒐𝒈 𝒂𝑪𝒖𝟐+ = 𝑬 𝒐 𝑪𝒖 − 𝐩𝑪𝒖 … … (𝟔. 𝟐)
𝟐 𝟐

Where, 𝒑𝑪𝒖 = −𝒍𝒐𝒈 𝒂𝑪𝒖𝟐+ 96

 i. Electrodes of the First Kind ….cont’d

 A general expression for any metal & its cation is

𝟎. 𝟎𝟓𝟗𝟐
𝐸𝑖𝑛𝑑 = 𝐸 𝑜 𝑋 𝑛+/ + 𝒍𝒐𝒈 𝒂𝑿𝒏+
𝑋 𝒏
𝟎.𝟎𝟓𝟗𝟐
= 𝐸 𝑜 𝑋 𝑛+/ − 𝒑𝑿
𝑋 𝒏

…………………..(6.3)

Where, 𝒑𝑿 = −𝒍𝒐𝒈 𝒂𝑿𝒏+

97
 i. Electrodes of the First Kind ….cont’d
 Electrode systems of the first kind are not widely used for
potentiometric determinations for several reasons.
 For one, metallic indicator electrodes are not very selective
& respond not only to their own cations but also to other
more easily reduced cations.
• For example, a copper electrode cannot be used for the
determination of copper(II) ions in the presence of silver(I)
ions because the electrode potential is also a function of
the Ag+ concentration. 9
8
 i. Electrodes of the First Kind ….cont’d
 In addition, many metal electrodes, such as Zn & Cd, can only be
used in neutral or basic solutions because they dissolve in the
presence of acids.
 Third, other metals are so easily oxidized that they can be used
only when analyte solutions are deaerated to remove oxygen.

 For these reasons, the only electrode systems of the first kind that
have been used in potentiometry are
 Ag/Ag+ & Hg/Hg2+ in neutral solutions &
9
 Cu/Cu2+, Zn/Zn2+, Cd/Cd2+, Bi/Bi3+, & Pb/Pb2+ in deaerated solns.
9
 ii. Indicator Electrodes of the Second Kind
 Metals not only serve as indicator electrodes for their own cations
but also respond to

 the activities of anions that form sparingly soluble precipitates or

 a different cation that form stable complexes (e.g. Ca complex with
EDTA)
E.g. The potential of a silver electrode, correlates reproducibly with the
activity of 𝐶𝑙− ion in a solution saturated with AgCl.

• In this situation, the electrode reaction can be written as

𝑨𝒈𝑪𝒍 𝒔 + 𝒆− ↔ 𝑨𝒈 𝒔 + 𝑪𝒍−(𝒂𝒒) 𝑬𝒐 𝑨𝒈𝑪𝒍/𝑨𝒈 = 𝟎. 𝟐𝟐𝟐 𝑽
1
The Nernst expression for this process at 25°C is
0
0
 ii. Indicator Electrodes of the Second Kind….cont’d

𝑬𝒊𝒏𝒅 = 𝑬𝒐 𝑨𝒈𝑪𝒍/𝑨𝒈 − 𝟎. 𝟎𝟓𝟗𝟐 𝒍𝒐𝒈 𝒂𝑪𝒍−

= 𝑬𝒐 𝑨𝒈𝑪𝒍/𝑨𝒈 + 𝟎. 𝟎𝟓𝟗𝟐𝐩𝑪𝒍 …………………(6.4)

 Eqn 6-4 shows that the potential of a Ag electrode is proportional

to pCl, the negative logarithm of the chloride ion activity.

 Thus, in a solution saturated with AgCl, a Ag electrode can serve

as an indicator electrode of the second kind for chloride ion.

 Note that the sign of the log term for an electrode of this type is
1
opposite that for an electrode of the first kind.
0
1
 iii. Inert Metallic Electrodes for Redox Systems

 Are relatively inert conductors respond to redox systems.

 Such materials as Pt, Au, Pd, & carbon can be used to monitor
redox systems.
• E.g. The potential of a Pt electrode immersed in a solution
containing cerium(III) & cerium(IV) is

𝒂𝑪𝒆𝟑+
𝐸𝑖 𝑛 = 𝐸 𝑜 𝐶𝑒 4+ − 𝟎. 𝟎𝟓𝟗𝟐 𝒍𝒐𝒈
𝑑
/𝐶𝑒3+ 𝒂𝑪𝒆𝟒+
• A Pt electrode is a convenient indicator electrode for titrations
involving standard cerium(IV) solutions. 1
0
2
 2. Membrane Indicator Electrodes

 Membrane electrodes are sometimes called p-ion electrodes

because the data obtained from them are usually presented as
p-functions, such as pH, pCa, or pNO3.

Examples of Membrane Indicator Electrodes

 The Glass Electrode for Measuring pH
 Glass Electrodes for Other Cations
 Liquid-Membrane Electrodes
1
 Crystalline-Membrane Electrodes
0
3
The Glass Electrode for Measuring pH
 It is the thin glass membrane bulb at the tip of the electrode that
responds to pH.

 Involves measurement of the potential that appears across a thin

glass membrane.

Glass Electrodes for Other Cations

 Glass electrodes that permit the direct potentiometric measurement

of univalent cations such as Na+, K+, NH4+, Rb+, Cs+, Li+, & Ag+.
 Some of these glasses are reasonably selective toward particular
1
singly charged cations.
0
4
Liquid-Membrane Electrodes

 The potential of liquid-membrane electrodes develops across the

interface between the solution containing the analyte & a liquid-
ion exchanger that selectively bonds with the analyte ion.

 These electrodes have been developed for the direct

potentiometric measurement of numerous cations (NH4+, Cd2+,
Ca2+, K+, water hardness (Ca2+ + Mg2+)) as well as certain anions
(Cl-, NO3-, NO2-, ClO4-).
1
0
5
Crystalline-Membrane Electrodes
 The potential that develops across crystalline solid-state electrodes
is measured.

 A membrane with anionic sites is selectively responds toward

certain cations, while a membrane with cationic sites might be
expected to respond selectively toward anions.

For example:
Membranes prepared from pellets of silver halides have been used
successfully in electrodes for the selective determination of 𝐶𝑙−,
1
𝐵𝑟−, & 𝐼− ions. 0
6
 3. Ion-Sensitive Field Effect Transistors (ISFETs)

 ISFETs offer a number of significant advantages over membrane

electrodes including sharpness, small size, inertness toward harsh
environments, rapid response, & low electrical impedance.

 In contrast to membrane electrodes, ISFETs do not require

hydration before use & can be stored indefinitely in the dry state.

 Despite these many advantages, no ISFET-specific ion electrodes

appeared on the market.
1
0
7
6.4 Potentiometric Titrations
 we measure the potential of a suitable indicator
electrode as a function of titrant volume.

 provide data that are more reliable than data

from titrations that use chemical indicators.

 are particularly useful with colored or turbid

solutions & for detecting the presence of
unsuspected species.
Figure 6-4 Apparatus for a
potentiometric titration.1
08
6.4 Potentiometric Titrations …cont’d

 Potentiometric titrations offer additional advantages over direct

potentiometry.
 Because the measurement is based on the titrant volume that causes a
rapid change in potential near the equivalence point, potentiometric
titrations are not dependent on measuring absolute values of Ecell.
 This characteristic makes the titration relatively free from junction
potential uncertainties because the junction potential remains
approximately constant during the titration.
 Titration results, instead, depend most heavily on having a titrant of
accurately known concentration.
1
0
9
6.4.1 Detecting the End Point
Some of the several methods used to determine the
end point of a potentiometric titration are

i. A direct plot of cell potential as a function of reagent volume.

Visually estimating the inflection point in the steeply rising or falling

portion of the curve & take it as the end point.

ii. A plot of the first derivative data (∆E/∆V) as a function of the

average volume.
From the plot, the point of maximum hill
coincides with the equivalence point.
1
1
0
Example: Potentiometric Titration
Data for 2.433 mmol of Chloride
with 0.1000 M AgNO3

Figure 6-5 Titration of 2.433 mmol of chloride

with 0.1000 M AgNO3.
(a) Titration curve. (b) First-derivative curve.
111
 6.4.2 Applications of potentiometric titration

A. Neutralization titrations
All are through
 Determining dissociation constants
the measurement
B. Redox Titrations of cell potentials.
 Determining equilibrium constants

C. Precipitation titrations
 Determining solubility-product constants

D. Complex formation titration

 Determining formation constants 1
1
2
Example:

Calculate the dissociation constant KHP for the weak acid HP if the cell

𝐒𝐂𝐄| 𝐇𝐏 𝟎. 𝟎𝟏𝟎 𝐌 , 𝐍𝐚𝐏 𝟎. 𝟎𝟒𝟎 𝐌 Pt, 𝐇𝟐 (𝟏. 𝟎𝟎 𝐚𝐭𝐦)

develops a potential of -0.591 V.

Solution
The diagram for this cell indicates that the saturated calomel electrode
is the left-hand electrode. Thus,

𝑬𝒄𝒆𝒍𝒍 = 𝑬𝒓𝒊𝒈𝒉𝒕 − 𝑬𝒍𝒆𝒇𝒕 = 𝑬𝒓𝒊𝒈𝒉𝒕 − 𝟎. 𝟐𝟒𝟒 = −𝟎. 𝟓𝟗𝟏 𝐕

11
𝑬𝒓𝒊𝒈𝒉𝒕 = −𝟎. 𝟓𝟗𝟏 + 𝟎. 𝟐𝟒𝟒 = −𝟎. 𝟑𝟒𝟕 𝑽
3
We then apply the Nernst equation for the hydrogen electrode to
find that

𝟎. 𝟎𝟓𝟗𝟐 𝟏. 𝟎𝟎
−𝟎. 𝟑𝟒𝟕 = 𝟎. 𝟎𝟎𝟎 − 𝒍𝒐𝒈
𝟐 [𝑯𝟑 𝑶+]𝟐

𝟐 𝒙 𝟎. 𝟎𝟓𝟗𝟐
= 𝟎. 𝟎𝟎𝟎 + 𝒍𝒐𝒈[𝑯𝟑 𝑶+]
𝟐
−𝟎. 𝟑𝟒𝟕 − 𝟎. 𝟎𝟎𝟎
𝒍𝒐𝒈[𝑯𝟑 𝑶+ ] = = −𝟓. 𝟖𝟔
𝟎. 𝟎𝟓𝟗𝟐
[𝑯𝟑 𝑶+] = 𝟏. 𝟑𝟖 𝒙 𝟏𝟎−𝟔
1
1
4
 By substituting this value of the [𝑯𝟑𝑶+] as well as the
concentrations of the weak acid & its conjugate base into the
dissociation constant expression, we obtain

[𝑯𝟑𝑶+][𝑷−] (𝟏. 𝟑𝟖 𝒙 𝟏𝟎−𝟔 )(𝟎. 𝟎𝟒𝟎)

𝑲𝑯𝑷 = = = 𝟓. 𝟓 𝒙 𝟏𝟎−𝟔
[𝑯𝑷] 𝟎. 𝟎𝟏𝟎

11
5
 UNIT-7
VOLTAMMETRY

 The term voltammetry refers to a group of electroanalytical

methods in which we acquire information about the analyte by
measuring current in an electrochemical cell as a function of
applied potential.

 When current proportional to analyte concentration is

monitored at a fixed potential, the technique is called
amperometry.

 We obtain this information under conditions that promote

polarization of a small indicator or working electrode.
116
 7.1 Excitation signals in voltammetry

 In voltammetry, a variable potential excitation signal is impressed

on a working electrode in an electrochemical cell.

 This excitation signal produces a characteristic current response,

which is the measurable quantity.

 The waveforms of four of the most common excitation signals

used in voltammetry are shown in Figure 7-1 below.

117
Figure7-1 Voltage versus time excitation signals used in voltammetry.

119
 The classical Voltammetric excitation signal is the linear scan in

which the voltage applied to the cell increases linearly (usually

over a 2- to 3-V range) as a function of time.

 The current in the cell is then recorded as a function of time &

thus as a function of the applied voltage.

 In amperometry, current is recorded at fixed applied voltage.

120
 Currents are measured at various times during the lifetime of these
pulses.

 With the triangular waveform, the potential is cycled between two

values, first increasing linearly to a maximum & then decreasing
linearly with the same slope to its original value.
 This process may be repeated numerous times as the
current is recorded as a function of time.

To the right of each of the waveforms of Figure 23-1 is listed the types
of voltammetry that use the various excitation signals.

121
 7.2 Voltammetric Instrumentation

The components of a simple

apparatus for carrying out
voltammetric measurements.

The cell is made up of three electrodes

immersed in a solution containing the
analyte and also an excess of a nonreactive
electrolyte called a supporting electrolyte.

122
Working electrode:
 is the electrode at which the analyte is oxidized or reduced.
 is microelectrode whose potential is varied linearly with time.

Reference electrode:
 has a potential that remains constant throughout the experiment.
 Ag/AgCl electrode or calomel.

Counter electrode:
 is often a coil of Pt wire or a pool of Hg that completes circuit.
 conducts e- from signal source through soln to the working electrode.

Supporting electrolyte:
 is a salt added in excess to the analyte soln to conduct current.
 most commonly, it is an alkali metal salt that does not react at the
working electrode at the potentials being used.
123
 7.3 Types of voltammetry

Hydrodynamic voltammetry is a type Cyclic voltammetry (CV) is performed

of voltammetry in which the analyte by cycling the potential of a working
solution is kept in continuous motion. electrode, & measuring the resulting
current.
Pulse voltammetry is a voltammetry
method used to make electrochemical
Polarography is the branch of
measurements & a derivative of linear
voltammetry in which a DME is used as
sweep voltammetry or staircase
the indicator electrode.
voltammetry, with a series of regular
voltage pulses superimposed on the
potential linear sweep or stair steps.

124
 7.4 Polarography and Amperometry

Polarography:

- is the branch of voltammetry in which a DME is used as the

indicator electrode.

- is the first type of voltammetry that was invented by the

Czechoslovakian chemist Jaroslav Heyrovsky.

- differs from other types of voltammetry in that the working

electrode is the unique DME.

125
 It occurs as the surface concentration
Polarographic Currents & Polarogram of the analyte falls to zero.

 Polarographic Currents are controlled by

diffusion alone.

 Diffusion current is the current observed

in polarography when the current is
limited only by the rate of diffusion.
• The diffusion current is proportional to the
concentration of analyte.

 The residual current is the small current observed

in the absence of an electro active species.
A graph of current vs. potential in a
 The limiting current is the current plateau that is polarographic experiment is called
observed at the top of the voltammetric wave. a polarogram.

126
 7.5 Applications of voltammetry

Organic Voltammetric Analysis Inorganic Applications

Voltammetry is applicable to the analysis of
The following organic functional
many inorganic substances.
groups produce voltammetric waves:
For examples:
1. Carbonyl groups  Most metallic cations are reduced at common
2. Certain carboxylic acids working electrodes.

3. Most peroxides & epoxides  Even the alkali & alkaline-earth metals are

4. Nitro, amine oxide, & azo groups reducible, provided the supporting electrolyte
does not react at the high potentials required;
5. Most organic halogen groups
in this instance, the tetra alkyl ammonium
6. Carbon/carbon double bonds
halides are useful electrolytes because of their
7. Hydroquinones.
high reduction potentials.

127
 Amperometry

Voltammetric methods are based on When current proportional to analyte

measuring current as a function of concentration is monitored at a fixed potential,
potential applied to a small electrode. the technique is called amperometry.

Since the potential is not scanned, amperometry does not

lead to a voltammogram.

One important application of amperometry is in the assembly of chemical sensors.

One of the first amperometric sensors to be developed was for dissolved O2 in blood.

128
 The design of the amperometric sensor is similar to potentiometric
membrane electrodes.

 A gas-permeable membrane is stretched across the end of the sensor & is

separated from the working & counter electrodes by a thin solution of
KCl.

 The working electrode is a Pt disk cathode, and an Ag ring anode is the

counter electrode.

 Although several gases can diffuse across the membrane (O2, N2, CO2),
only O2 is reduced at the cathode.

128
Clark amperometric
Sensor for the Determination of
Dissolved O2

130
 UNIT-8
COULOMETRY AND ELECTROGRAVIMETRIC ANALYSIS

 Electrogravimetry and coulometry are related methods that are based on

electrolysis carried out long enough to ensure complete oxidation or
reduction of the analyte to a product of known composition.

 In electrogravimetry, the goal is to determine the amount of analyte

present by converting it to a product that is weighed as a deposit on one of
the electrodes.

 In coulometry, we determine the amount of analyte by measuring the

quantity of electrical charge needed for complete conversion to a product.

131
 8.1 Coulometric Methods

8.1.1 Determining the Electrical Charge (𝑸)

 Electrical charge is the basis of the other electrical quantities

(current, voltage, & power).

 The charge Q that results from a constant current of I amperes

operated for t seconds is

𝑸 = 𝑰𝒕 … … … … … … (𝟖. 𝟏)

 For a variable current i, the charge is given by the integral

𝑡
𝑸 = ∫0 𝒊 𝑑𝑡 …………… (8.2)
131
The faraday (F ) is the quantity of charge that corresponds to one mole or
6.022 x 1023 electrons.
Since each 𝑒− has a charge of 1.6022 x 10-19 C, the faraday also equals
96,485 C. 6.022 𝑥 1023𝑒− 1.6022 𝑥 10−19𝐶
𝐹= 𝑥 = 𝟗𝟔, 𝟒𝟖𝟓 𝑪/𝐦𝐨𝐥
1 𝑚𝑜𝑙 𝑒− 1 𝑒−

Faraday‘s law relates the number of moles of the analyte 𝑛𝐴, to the charge Q

𝑸
𝒏𝑨 = … … … … … (𝟖. 𝟑)
𝒏𝑭

where n is the no. of moles of electrons in the analyte half-rxn.

∴ We can use these definitions to calculate the mass of a chemical species

that is formed at an electrode by a current of known magnitude.

132
Example 8-1
A constant current of 0.800 A is used to deposit copper at the cathode and
oxygen at the anode of an electrolytic cell. Calculate the number of grams of

each product formed in 15.2 min, assuming no other redox reaction occurs.

Solution
The two half-reactions are
𝐶𝑢2+ + 2𝑒− → 𝐶𝑢 𝑠
2𝐻2𝑂 → 4𝑒− + 𝑂2 𝑔 + 4𝐻+

Thus, 1 mol of copper is equivalent to 2 mol of 𝑒−s, & 1 mol of oxygen

corresponds to 4 mol of 𝑒−s.

134
 Substituting into Equation 8-1 yields

60 𝑠
𝑄 = 0.800 𝐴 𝑥 15.2 min 𝑥 = 729.6 𝐴. 𝑠 = 𝟕𝟐𝟗. 𝟔 𝑪
𝑚𝑖𝑛

We can find the number of moles of Cu & O2 from Eqn 8-3

729.6 𝐶 −𝟑 𝒎𝒐𝒍 𝑪𝒖
𝑛𝐶𝑢 = = 𝟑. 𝟕𝟖𝟏 𝒙 𝟏𝟎
2 𝑚𝑜𝑙 𝑒−/ 𝑥 96,485 𝐶/
𝑚𝑜𝑙 𝐶𝑢 𝑚𝑜𝑙 𝑒−

729.6 𝐶
𝑛 𝑂2 = = 𝟏. 𝟖𝟗𝟎 𝒙 𝟏𝟎−𝟑 𝒎𝒐𝒍 𝑶𝟐
4 𝑚𝑜𝑙 𝑒−/ 𝑥 96,485 𝐶/
𝑚𝑜𝑙 𝑂2 𝑚𝑜𝑙 𝑒−

135
The masses of Cu & O2 are given by

63.55 𝑔 𝐶𝑢
𝑚𝑎𝑠𝑠 𝐶𝑢 = 𝟑. 𝟕𝟖𝟏 𝒙 𝟏𝟎−𝟑 𝒎𝒐𝒍 𝑥 = 𝟎. 𝟐𝟒𝟎 𝒈 𝑪𝒖
𝑚𝑜𝑙

32.00 𝑔 𝑶𝟐
𝑚𝑎𝑠𝑠𝑶𝟐 = 𝟏. 𝟖𝟗𝟎 𝒙 𝟏𝟎−𝟑 𝒎𝒐𝒍 𝒙 = 𝟎. 𝟎𝟔𝟎𝟓 𝒈 𝑶 𝟐
𝑚𝑜𝑙

136
8.1.2 Types of Coulometric Methods

Two methods have been developed that are based on measuring the

quantity of charge:

i. controlled-potential coulometry &

ii. controlled-current coulometry

137
i. Controlled-Potential (potentiostatic) Coulometry:

- The potential of the working electrode is maintained at a constant level

such that only the analyte is responsible for conducting charge across the
electrode/solution interface.

=> It is performed with a constant potential source

- The charge required to convert the analyte to its reaction product is then
determined by recording and integrating the current-versus-time curve
during the electrolysis.
𝒕
𝑸 = ∫ 𝒊 𝒅𝒕
𝟎

138
Instrumentation for potentiostatic coulometry :

The instrumentation for potentiostatic coulometry consists of

 an electrolysis cell (assembled with 3 electrodes, solution holder

 Working electrode can be platinum-gauze
& stirrer), or mercury-pool.
 Reference electrode can be SCE
 a potentiostat, &

 a device for determining the charge consumed by the analyte

(Coulometer)

139
Electrolysis cells for potentiostatic coulometry.
140
Example 8-2

The Fe(III) in a 0.8202-g sample was determined by coulometric reduction

to Fe(II) at a platinum cathode. Calculate the percentage of Fe2(SO4)3 (M =

399.88 g/mol) in the sample if 103.2775 C were required for the reduction.

141
Solution
Since 1 mol of 𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 consumes 2 mol of 𝒆−s, we may write from Eqn 8-3

𝟏𝟎𝟑. 𝟐𝟕𝟕𝟓 𝑪
𝒏𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 =
𝟐 𝒎𝒐𝒍 𝒆−/ 𝒙 𝟗𝟔, 𝟒𝟖𝟓 𝑪/
𝒎𝒐𝒍𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 𝒎𝒐𝒍 𝒆−

= 𝟓. 𝟑𝟓𝟐𝟎 𝒙 𝟏𝟎−𝟒 𝒎𝒐𝒍 𝑭𝒆𝟐 (𝑺𝑶𝟒 )𝟑

399.88 𝑔𝑭𝒆𝟐 (𝑺𝑶𝟒 )𝟑

𝑚𝑎𝑠𝑠 𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 = 𝟓. 𝟑𝟓𝟐𝟎 𝒙 𝟏𝟎−𝟒 𝒎𝒐𝒍 𝒙
𝑚𝑜𝑙 𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑

= 𝟎. 𝟐𝟏𝟒𝟎𝟏 𝒈 𝑭𝒆𝟐 (𝑺𝑶𝟒 )𝟑

Percentage 𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 = 𝟎.𝟐𝟏𝟒𝟎𝟏 𝒈𝑭𝒆𝟐(𝑺𝑶𝟒)𝟑 𝒙 𝟏𝟎𝟎% = 𝟐𝟔. 𝟎𝟗%

𝟎.𝟖𝟐𝟎𝟐 𝒈 𝒔𝒂𝒎𝒑𝒍𝒆

142
ii. Controlled-current (Amperostatic) coulometry:

- is also called coulometric titrimetry.

- is performed with a constant-current source, sometimes called galvanostat.
- is a method in which the amount of analyte is determined from the
magnitude of the current & the time required to complete the titration.
- is similar to volumetric titrimetry in that analyses are based on measuring
the combining capacity of the analyte with a standard reagent.

 The magnitude of the current in amperes is analogous to the molar

concentration of a standard solution, & the time measurement is analogous
to the volume measurement in conventional titrimetry.

 A coulometric titration offers several significant advantages over a

volumetric procedure. 142
Advantages of Coulometric Titrations over Volumetric Titrations
Electrons are the reagent
Coulometric titrations:
in a coulometric titration.
- are as accurate & precise as comparable volumetric methods.

- eliminate the problems associated with the preparation, standardization, & storage
of standard solutions In a coulometric determination is, however, straightforward because
they are consumed as soon as they are generated.

- also best when small amounts of analyte have to be titrated

 Electrons are added to the analyte (via

- provides reagents for precipitation, complex formn,
the direct current) or to some species
neutralization, or oxidation/reduction titrations that immediately reacts with the
analyte until an end point is reached.
- are more readily automated since it is easier
 At an end point, the electrolysis is
to control electrical current than liquid flow discontinued.

144
  At an end point , the
End Point Detection in Coulometric titrations
electrolysis is discontinued.

The end point in coulometric titrations can be detected by

a. Color indicators (like their volumetric counterparts)

 Generally, the end point detection methods in volumetric

methods are applicable to coulometric titrations as well.

 E.g. For the titration of Fe(II), a redox

indicator, like 1,10-phenanthroline, can
be used
(𝒑𝒉𝒆𝒏)𝟑𝑭𝒆𝟐+ ↔ (𝒑𝒉𝒆𝒏)𝟑𝑭𝒆𝟑+ + 𝒆−
Ferroin
𝒓𝒆𝒅 𝒑𝒂𝒍𝒆 𝒃𝒍𝒖𝒆

b. Potentiometrically (as an alternative method)

145
Instrumentation in Amperostatic coulometry:
Constant-current generators are
The equipment required for
sometimes called galvanostats.
a coulometric titration includes:

 a source of constant current

 a titration cell (consists of WE & CE)
CE is sintered with glass disk to isolate from
the rxn medium & prevent interference by
the rxn products from this electrode.

 a timer, &
 a device for monitoring current.
(Current meter)

 A typical coulometric titration cell consisting of a

Conceptual diagram
working electrode at which the reagent is produced & a
counter (auxiliary) electrode to complete the circuit. of a coulometric titration apparatus.

146
  The working electrode used to
generate reactants in situ is often
referred to as the generator electrode.
It is usually a platinum rectangle, a
coil of wire, or a gauze cylinder with a
relatively large surface area to
minimize polarization effects.

 The counter electrode is usually

isolated from the reaction medium by
a sintered disk or other porous
medium to prevent interference by the
reaction products from this electrode.

A typical coulometric titration cell.

147
Applications of Coulometry

 Applications of potentiostatic coulometry:

- used to determine more than 55 elements in inorganic compounds.

- used in the nuclear energy field for the relatively interference-free

determination of uranium & plutonium.

- also offers possibilities for the electrolytic determination (&

synthesis) of organic compounds.

E.g. Trichloroacetic acid is quantitatively reduced at a mercury cathode whose potential is
suitably controlled:

𝑪𝒍𝟑𝑪𝑪𝑶𝑶− + 𝑯+ + 𝟐𝒆− → 𝑪𝒍𝟐𝑯𝑪𝑪𝑶𝑶− + 𝑪𝒍−

148
 Applications of Coulometric titrations :

Coulometric titrations have been developed for all types of

volumetric rxns.

 Neutralization Titrations

 Precipitation & Complex-formation reactions

 Oxidation/reduction Titrations

149
 Types of Electrogravimetric Methods

There are two general types of electrogravimetric methods.

i. Constant current electrogravimetry

 Here, electrodeposition is carried out by keeping

the current constant & periodically increasing
the applied potential as the electrolysis proceeds.

 uses simple & inexpensive equipment.

 require little operator attention.

150
Instrumentation for Constant current electrogravimetry

The apparatus for constant current

electrogravimetry consists of

 a suitable cell (a two-electrode cell)

 a direct current source

 An ammeter

Apparatus for electrodeposition of metals

 Voltmeter without cathode-potential control.

151
ii. Controlled-potential (potentiostatic) electrogravimetry

 Here, the potential of the WE is maintained at a constant level.

 A potentiostat automatically maintains the WE potential at a

constant value relative to a reference electrode, like SCE.

151
 8.2 The effect of current on cell potential

Two phenomena are must be considered:

Ohm's law states that the current through a
i. IR drop &
conductor between two points is directly
ii. Polarization proportional to the voltage across the two points.
i = E/R

1. Ohmic Potential: IR Drop The product of the resistance R of a cell

in ohms (𝛺) & the current I in amperes
Ohm’s law: E = IR, or I = E/R. (A) is called the Ohmic potential or the
IR drop of the cell.

The units of R are ohms (𝛺). One ohm equals one volt per ampere (1𝜴 = 1V/A).
Thus, the product IR has the units of amperes x volts/ampere = volts.

153
 In order to generate a current of I amperes in this cell, we must
apply a potential that is IR volts more negative than the
thermodynamic cell potential,

𝑬𝒄𝒆𝒍𝒍 = 𝑬𝒓𝒊𝒈𝒉𝒕 − 𝑬𝒍𝒆𝒇𝒕, 𝒕𝒉𝒂𝒕 𝒊𝒔

𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 = 𝑬𝒄𝒆𝒍𝒍 − 𝑰𝑹 … … … … … . (𝟖. 𝟒)

Resistance = opposition to electric current

Usually we try to minimize the IR drop in the cell by

 having a very small cell resistance (high ionic strength) or

 using a special three-electrode cell in which the current passes b/n
the WE & an auxiliary or CE.

154
Example 8-3: The following cell has been used for the determination
of cadmium in the presence of 𝐶𝑙− ions by both
electrogravimetry & coulometry:

𝑨𝒈|𝑨𝒈𝑪𝒍 𝒔 , 𝑪𝒍− 𝟎. 𝟐𝟎𝟎 𝑴 , 𝑪𝒅𝟐+ 𝟎. 𝟎𝟎𝟓𝟎𝟎 𝑴 |𝑪𝒅

An electrolytic cell for determining Cd2+.

(a) Current = 0.00 mA. (b) Schematic of
cell in (a) with internal resistance of cell
represented by a 15.0- Ω resistor

& 𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 increased to give a current of

2.00 mA.
155
Calculate the potential that

a. must be applied to prevent a current from developing in the cell

when the two electrodes are connected &

b. must be applied to cause an electrolytic current of 2.00 mA to

develop. Assume that the internal resistance of the cell is 15.0 Ω.

Use the following standard reduction potentials:

𝑪𝒅𝟐+ + 𝟐𝒆− ↔ 𝑪𝒅 𝒔 𝑬𝒐 = −𝟎. 𝟒𝟎𝟑 𝑽

𝑨𝒈𝑪𝒍 𝒔 + 𝒆− ↔ 𝑨𝒈 𝒔 + 𝑪𝒍− 𝑬𝒐 = 𝟎. 𝟐𝟐𝟐 𝑽

156
Solution:
The potential of the cadmium electrode (𝑬𝒓𝒊𝒈𝒉𝒕) is

𝟎. 𝟎𝟓𝟗𝟐 𝟏
𝑬𝒓𝒊𝒈𝒉𝒕 = −𝟎. 𝟒𝟎𝟑 − 𝐥𝐨𝐠 = −𝟎. 𝟒𝟕𝟏 𝑽
𝟐 𝟎. 𝟎𝟎𝟓𝟎𝟎

& that of the silver electrode (𝑬𝒍𝒆𝒇𝒕) is

𝑬𝒍𝒆𝒇𝒕 = 𝟎. 𝟐𝟐𝟐 − 𝟎. 𝟎𝟓𝟗𝟐 𝒍𝒐𝒈(𝟎. 𝟐𝟎𝟎) = 𝟎. 𝟐𝟔𝟑 𝑽

Since the current is to be 0.00 mA, we find from Eqn 8.4,

𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 = 𝑬𝒄𝒆𝒍𝒍 = 𝑬𝒓𝒊𝒈𝒉𝒕 − 𝑬𝒍𝒆𝒇𝒕

= −𝟎. 𝟒𝟕𝟏 𝐕 − 𝟎. 𝟐𝟔𝟑 𝐕 = −0.734 V

157
• To prevent the passage of current in this cell, we would need to

apply a voltage of −0.734 V, as shown in Figure 8-1a.

• Note that in order to obtain a current of 0.00 mA (null point), the

applied voltage must exactly match the galvanic cell potential.

At null point, the standard voltage is read on a

voltmeter to obtain the value of 𝑬𝒄𝒆𝒍𝒍.

158
b. In order to calculate the applied potential needed to develop a current

of 2.00 mA, or 2.00 x 10-3 A, we substitute into Eqn 8.4 to give

𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 = 𝑬𝒄𝒆𝒍𝒍 − 𝑰𝑹
= −𝟎. 𝟕𝟑𝟒 𝑽 − 𝟐. 𝟎𝟎 𝒙 𝟏𝟎−𝟑 𝑨 𝒙 𝟏𝟓 𝛺

= −𝟎. 𝟕𝟑𝟒 𝑽 − 𝟎. 𝟎𝟑𝟎 𝑽 = −𝟎. 𝟕𝟔𝟒 𝑽

 We see that to obtain a 2.00 mA current as in Figure 8-1b, an

applied potential of −𝟎. 𝟕𝟔𝟒 𝑽 is required.

159
2. Polarization Effects

If we solve Eqn 8.4 for the current I, we obtain

𝑬𝒄𝒆𝒍𝒍 − 𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 𝑬𝒄𝒆𝒍𝒍

𝑰= =− + … … … … … (𝟖. 𝟓)
𝑹 𝑹 𝑹

 Note that a plot of current I in an electrolyte cell versus 𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅

should be a straight line with a slope equal to the negative
reciprocal of the resistance, -1/R, & an intercept equal to 𝑬𝒄𝒆𝒍𝒍/R.

Consider Figure 8-2 below.

160
 -0.839 V
Theoretical curve
assuming no polarization.

Overvoltage

Cells that exhibit nonlinear behavior at higher

currents exhibit polarization, & the degree of
polarization is given by an overpotential (∏).

Experimental current/voltage curve for operation of the cell

161
Polarization:
- is the departure of the electrode potential from its theoretical Nernst eqn
value on the passage of current.
- is an electrode phenomenon that may affect either or both of the electrodes
in a cell, & varies widely.
- requires the application of a potential greater than the theoretical value to
give a current of the expected magnitude.

∴ For an electrolytic cell affected by overvoltage, Eqn 8-4 then becomes

𝑬𝒂𝒑𝒑𝒍𝒊𝒆𝒅 = 𝑬𝒄𝒆𝒍𝒍 − 𝑰𝑹 − ∏ … … … … … . 𝟖. 𝟔

Overvoltage (∏) - is the potential d/ce b/n the theoretical 𝑬𝒄𝒆𝒍𝒍 from
Eqn 8-4 & the actual 𝑬𝒄𝒆𝒍𝒍 at a given level of current.
162
Factors that influence polarization are

1. Electrode size, shape, & composition;

2. Composition of the electrolyte solution;
3. Temperature & stirring rate;
4. Current level; &
5. Physical state of the species participating in the cell rxn.

 Polarization phenomena can be divided into two categories:

1. concentration polarization &

2. kinetic polarization.

162
Concentration polarization:
- occurs when reactant species do not arrive at the surface of the electrode
or product species do not leave the surface of the electrode fast enough to
maintain the desired current.

 When these events happen, the current is limited to

values less than that predicted by Eqn 8-4.

 Reactants are transported to & products away from an electrode

by three mechanisms: i. Diffusion
ii. Migration, and
iii. Convection.

164
Diffusion
- is the movement of a species under the influence of a concentration gradient.
- is the process that causes ions or molecules to move from a more
concentrated part of a solution to a more dilute.
The rate of diffusion is directly proportional to the concentration difference.

 As Cd2+ are reduced to Cd atoms at the electrode

surface, the conc. of Cd2+ at the surface becomes
smaller than the bulk concentration.

 Cd2+ ions then diffuse from the bulk of the soln

to the surface as a result of the conc. gradient.

Diffusion of Cd2+ ions from the bulk of the solution to the

electrode surface.
165
Migration
- is the movement of ions through a solution as a result of electrostatic
attraction between the electrodes & the ions.
- causes anions to be attracted to the positive electrode & cations to the
negative electrode.

Migration – Motion arising due to the

influence of an external field.

Convection
- is the transport of ions or molecules
The motion of ions through a solution because of
through a solution as a result of the electrostatic attraction between the ions & the
stirring, vibration, or temperature electrodes is called migration.

gradients.

166
Kinetic Polarization:

- is most commonly encountered when the reactant or product in an

electrochemical cell is gas.

- can be negligible for deposition or dissolution of such metals as Cu, Ag,

Zn, Cd, & Hg.

The current in a kinetically polarized cell is governed by the rate of

electron transfer rather than the rate of mass transfer.

 Mass transfer is the movement of material, such as ions, from one location
to another. => governs concentration polarization.

167
 UNIT-9
CONDUCTOMETRY
Basic principles and Instrumentation

 Conductometry: is the simplest of the electroanalytical

techniques.
 Conductometry is measuring the conductivity of ionic
solutions caused by mobility of ions towards respective
electrodes in presence of an electric field
 Conductivity: is the ability of the medium to carry electric
167
current.
 Conductivity is measured by using conductometer and its
unit is mhos(Ω-1)

C= 1 , 𝑅 = 𝑉, R-resistivity
𝑅
 Electrolyte solutions
𝑖
behave as an electrically conductive
medium.
 The change of electrolyte conductivity can indicate the
change of the concentration of the total ions, this method is
defined as conductometric Analysis

 Conductors: are either metallic or electrolytic.

169
Conductance of electricity: migration of positively charged
ions towards the cathode and negatively charged ones
towards the anode.

 Conductance depends on the number of ions in solution.

Factors affecting conductance
1. Temperature
 increase in temperature causes increase in conductance.
2. Nature of ions
 Size, molecular weight and number of charges.

170
3. Concentration of ions
 As the number of ions increases, the conductance
increases.

4. Size of electrodes
 Conductance is directly proportional to the cross
sectional area (A).
Specific conductance (K)
 Conductance is directly proportional to the cross section
area A and is inversely proportional to the length of a
uniform conductor. 170
 Thus,
G  A/l so G =KA/L
where K is the specific conductance

Conductivity measurements

 Two parallel platinized Pt foil electrodes or Pt

black with electrodeposited a porous Pt film which
increases the surface area of the electrodes and
further reduces faradaic polarization. 171
[Link] standard solutions
o Primary standard KCl solution

3. Conductivity Cell

 Avoid the change of temperature during determination

4. DC and AC Conductometry
 AC Conductometry can minimize the error caused by
electrode polarization

172
 Factors affecting conductivity

 Size of ions
 Temperature
 Number of ions
 Charge of ions
Specific conductivity:- is conductivity offered by a substance
of 1cm length and 1cm2 surface area.
 units are mhos/cm.
Equivalent conductivity:- is conductivity offered by a
solution containing equivalent weight of solute in it. 173
 APPLICATIONS OF CONDUCTOMETRY
1. Direct or absolute measurements

i. Determination of high purity water

 Conductivity is used to determine the purity of:
 drink water  deionized water and
 natural water  Wastewater
ii. Determination of the total concentration of strong
electrolyte solutions

 salinity of soil and sea water

174
iii. Determination of gaseous environmental contaminant in
air. eg. SO3, NO2, acidic rain
 determine the change of conductivity after absorption
2. Conductometric titrations
i. Very dilute solutions.
ii. Turbid and highly coloured solutions.
iii. Reaction which is not complete and where there is no
suitable indicator
CONDUCTOMETRIC TITRATIONS
 It the determination of end point of a titration by means of
175
conductivity measurements
 Types of conductometric titrations
 Acid-base titration
 Precipitation titration
 Replacement titration
 Redox (oxidation-reduction) titration
176
 Complexometric titration
1. ACID-BASE TITRATIONS

i. Titration of strong acid

(a) with strong base e.g. HCl with NaOH
(b) with weak base e.g. HCl with NH4OH

178
ii. Titration of weak acid

(c) with strong base e.g. CH3COOH with NaOH

(d) with weak base e.g. CH3COOH with NH4OH

179
2. PRECIPITATION TITRATIONS

K+ + Cl- → AgCl

180
3. REPLACEMENT TITRATIONS
[Link] of strong acid and weak base vs. strong base
Eg: ammonium chloride vs. sodium hydroxide
NH4Cl+NaOH→NH4OH+NaCl

181
B. Salt of strong base and weak acid vs. strong acid
eg. sodium acetate vs. hydrochloric acid

CH3COONa + HCl → CH3COOH + NaCl

182
4. REDOX TITRATION
Titration of ferrous ions with dichromate ions:
6Fe2+ + Cr2O72- + 14H+→ 6Fe3+ + 2Cr3+ +7H2O

183
 5. COMPLEXOMETRIC TITRATION
Eg. KCl vs. Hg(ClO4)2

 Non-aqueous titrations can also be measured

using conductometry.
Eg.

a)titration of weak bases vs. perchloric acid in

dioxan-formic acid.

183
 b)Titration of weak organic acids in methanol vs.
tetra methyl ammonium hydroxide in methanol-
benzene.
ADVANTAGES OF CONDUCTOMETRIC TITRATIONS

 No need of indicator
 Colored or dilute solutions or turbid suspensions
can be used for titrations.

 Temperature is maintained constant throughout

184
the titration.
 End point can be determined accurately and
errors are minimized as the end point is being
determined graphically.
DISADVANTAGES OF CONDUCTOMETRIC
TITRATIONS

 non specificity
 interference of high conc. of other electrolytes.

186
 UNIT-10
ELECTROPHORESIS
Principle
 The term electrophoresis describe the migration of a charged particle under
the influence of an electric field.
 Electrophoresis is a laboratory technique for separating mixtures of charged
molecules.
 It is a technique used to separate macromolecules in a fluid or gel based on
their charge, binding affinity, and size under an electric field.

Mixture: a material composed of two or more elements or parts.

Charged Molecules: a molecule that has too many or too few electrons.

187
 Many important biological molecules such as:
 amino acid,
 peptides,
 proteins,
 nucleotides and
 nucleic acids, possess ionisable groups and at any given pH, exist
in a solution as electrically charged species either as a cation (+) or
anion(-)

 Under the influence of an electric field these charged particles will

migrate either to cathode or anode, depending on the nature of their net
charge.

188
 Separation of a Mixture of Charged Molecules

 Charged molecules are separated based on their electrical charge and

size.

Positive Molecules

Analyze

Charge Size Identify

Separation Separation

Mixture of
Purify
Charged
Negative Molecules 188
Molecules
 The equipment required for electrophoresis consist basically of two
items:

power pack
Supplies a direct current between electrode in the electrophoresis unit.
electrophoresis unit
Available for running either vertical or horizontal Gel system.

189
vertical horizontal
COMPONENTS OF ELECTROPHORESIS

• Electrical Current – the flow of electric charge

• Positive Electrode – the wire that collects electrons
• Negative Electrode – the wire that emits electrons
• Porous – containing pores, permeable to fluids and small particles
• Sieve – a mesh device to filter small particles from larger particles.

 Anaphoresis is the electrophoresis of negative charge particles or anions

whereas cataphoresis is electrophoresis of positive charge ions or
cations.

190
 Types of Electrophoresis
1. Capillary electrophoresis
i. Gel electrophoresis

 Two kinds of gels are commonly used:

a. Agarose b. polyacrylamide
ii. Paper electrophoresis
2. Slab electrophoresis
i. Zone electrophoresis
ii. Immunoelectrophoresis
iii. Isoelectrofocusing

Application
 Electrophoresis has a wide application in separating and analysing
191
biomolecules such as proteins, plasmids, RNA, DNA, nucleic acids.
192