POLAROGRAPHY

VOLTAMMETRY

• Voltammetry is an electrochemical technique in which the

current(I) at an electrode is measured as a function of potential
(E) or voltage in an electrochemical cell. OR
• It is a method in which an external potential is applied in a
system to carry out electrochemical analysis.
• In short it is a study that involves current and voltage. It gives
information about the analyte.
• A minimum potential is required to initiate oxidation and
reduction reaction at an electrode.
 Voltammogram

• The change in
current with the
varying voltage
gives the plot and
is known as
voltammogram.

TYPES OF VOLTAMMETRY

• Linear sweep voltammetry

• Cyclic voltammetry
• Staircase voltammetry
• Normal pulse voltammetry
• Differential pulse voltammetry
• Polarography
 Depending on the choice of working electrode

• Depending on the choice of working electrode ,

the type of voltammetry is decided e.g.
• Dropping mercury electrode (DME) in
polarography
• Platinum electrode in cyclic voltammetry
• Glassy carbon in linear sweep voltammetry
• Current in the three electrode system is measured
in the range of microampere and miliampere.

POLAROGRAPHY

• Polarography is a branch of voltammetry in which the

information about the analyte can be obtained by
studying current-voltage curve.
• Polarography also called as a polarographic analysis is
an electrochemical method of analyzing solution of
reducible or oxidizable substances.
• Polarography was first invented by J. Heyrovsky
(Nobel Prize 1959). Differs from voltammetry in that
it employs a dropping mercury electrode (DME) to
continuously renew the electrode surface.
• In general it is a technique in which electric potential is
varied in a regular manner between 2 sets of electrodes
(indicator and reference) and current is monitored.
• This involve the electrolysis of a solution containing
electroactive species (usually a metal ion) so that
reduction takes place at the polarized electrode acting as a
cathode.
• Of the various microelectrodes available, the most
commonly used electrode is Dropping Mercury
Electrode(DME).

• The curves obtained by plotting the current flowing

through the electrolytic cell against applied voltage are
called current voltage curves. The graphical
representation of these curves are called polarograms.
• The apparatus used is known as polarograph and the
technique used is polarography.
• The electrolyte cell used is called polarographic cell.
• One of the virtues of polarography is that solutions as
dilute as 10-8 M can be analyzed and sample volumes
as small as 0.05 ml can be manipulated.
 In this technique the electrolysis of a
solution of an electroactive substance is
carried out in a polarographic cell.

PRINCIPLE OF
POLAROGRAPHY

This type of cell consists of a Dropping

Mercury Electrode acting as a cathode
and a reference electrode ( a pool of
mercury or saturated calomel electrode,
SCE) acting as anode.

IONS TRANSPORT IN
ELECTROCHEMICAL CELL
• Electroactive material can reach the surface of an electrode by three processes:
• Convection
• Migration
• Diffusion
 CONVECTION

• It refers to the
transport of ions
towards electrode due
to agitation,Vibrations
or difference in density
and temperature in
solution. (mechanical or
thermal disturbance)

MIGRATION

• It involves the
movement of the
oppositely charged
ions towards
electrode due to the
electrostatic
attractions.
 DIFFUSION

• Diffusion of particles
arises due to the
concentration
gradient of the
analyte between the
surface of electrode
and bulk of solution.

INSTRUMENTATION
• The basic components of the experiment set-up are
• A DC source of constant voltage (D)
• Voltage divider or potentiometer (P)
• Voltmeter (V)
• Galvanometer (G)
• Polarizable electrode (electrode potential has great changes
when small current flow through electrode e.g. DME)
• Non-polarizable electrode (If electrode potential doesn’t
change with the current e.g. SCE)
• Polarographic cell
INSTRUMENTATION
• The instrumentation of polarography consist of
a polarographic cell with electrodes.
• The electrodes are:
1. Cathode: Dropping mercury
electrode(DME), it is a working electrode.
2. Anode: Static calomel electrode (SCE), it is a
counter electrode.
3. Auxillary electrode: Pt wire
• The cell is connected in series with a
galvanometer (for measuring the flow of
current) in an electrical circuit that also contains
a battery or other source of direct current and a
device for varying the voltage.

POLAROGRAPHIC ELECTROCHEMICAL CELL

It is a H shaped vessel, the two limbs of

which are connected through an agar-agar
salt bridge (B) and a sintered glass disc
(d). One half of cell acts as a anode (A)
which is usually a reference electrode
such as saturated calomel electrode
(SCE). The other half of the cell contains
the solution of the sample , analyte (a)
and the Dropping Mercury electrode
(DME) serves as a cathode (C).
Salt bridge; glass tube filled with jelly
like substance like agar mixed with
electrolyte like KCl, KNO3 etc
 DROPPING MERCURY ELECTRODE
• Description of DME:
The DME consists of a 8-30 cm long capillary
tube, with an inner diameter of 0.05-0.1mm that is
vertically connected to the mercury reservoir (R)
through the connector. Electrical connection is
made by a wire dipping into the mercury in the
reservoir.
Under this condition, a continuous series of highly
reproducible spherical drops of mercury are
formed having diameter of 0.1-1mm.
As mercury flows through the capillary, drops
grow at the tip until they become heavy enough to
get detached from the tube. The typical drop time
is in the range of 2-6 seconds.
 WORKING
• The solution to be analyzed is placed in a glass cell containing two electrodes. At
first the nitrogen/hydrogen is passed through the electrolytic solution to remove
the dissolved oxygen.
• The DME is connected to the negative terminal of the DC source while the SCE
is connected to the positive terminal of the DC source. By suitably controlling the
potentiometer any EMF upto 3 V may be gradually applied to the cell.
• The voltage is increased by small increment and the corresponding current is
observed on the galvanometer. The current is very small until the applied voltage
is increased to a value large enough to cause the substance being determined to be
reduced at DME.
• The current increases rapidly at first as the applied voltage is increased above
critical value but gradually attains the a limiting value and remains more or less
constant as the voltage is further increased.
 The constant renewal of the electrode surface

ADVANTAGES
OF DME
Each drop has a smooth and uncontaminated surface
free from any adsorbed analyte or impurity

Rapid constant current achievement

Mercury forms amalgam (solid solution with many

metals). Amalgam formation lowers the activity of
the metal, thereby facilitating the reduction of the
metal ion.

DISADVANTAGES

The capillary may be easily

Oxidation of mercury itself to plugged so care must be taken
mercurous ion at potentials to avoid touching the tip of
more positive than +0.4 V. capillary with foreign material
especially fingers.

Applied voltage produces

The mercury is volatile and changes in surface tension and
toxic therefore safety hence change in drop size
precautions must be taken. resulting in oscillations in
galvanometer readings.
 POLAROGRAM
• A graph of current versus potential is called a polarogram
(sigmoid shaped).Concentration of electroactive species can
be determined.
Type of current represented
1. Residual current
2. Migration current
3. Diffusion current
4. Limiting current

Residual current
• Whenever an electrode comes into contact with an electrolyte solution,
there is an electrode-electrolyte interface (a double layer).
• When a voltage is applied this layer behaves like two plates of capacitor, a
small current will flow to charge them. Small current will also result due
to the reduction of small heavy metals which may be incorporated as
impurities in distilled water while preparation of an electrolyte solution
called Residual current. Potential applied is called decomposition
potential.
Diffusion current
• It is the current resulting from the diffusion of electroactive species
to the drop surface. Increase in volt is accompanied by increase in
current.
• It is due to the actual diffusion of electro reducible ions from the
bulk of the sample to the surface of the mercury droplet due to
concentration gradient.

Limiting current
Beyond a certain potential, the current reaches a steady
value called as limiting current.
At this point the rate of diffusion is equal to the rate of
reduction (saturation stage).
HALF WAVE POTENTIAL
• The half-wave potential is unique for each element and its different
valence states and chemical forms.
• Half wave potential is important for the analysis of ions present in
the analyte. As half wave potential is fix for certain cation or
functional group.

APPLICATIONS OF POLAROGRAPHY
• Quantitative analysis of dissolved oxygen:
• Used for determination of quantitative analysis of dissolved
oxygen in wide range of fluids including whole blood, fluids,
milk etc. oxygen dissolved in an electrolyte solution gets
reduced at DME.

• Trace metals and metal containing drugs:

• Used to determine the trace metals in pharmaceutical products
and to estimate the drugs that contain metals as a constituent.
The metals examined include antimony, iron, copper, zinc,
magnesium etc.
• Estimation of vitamins:
• Most water soluble or oil soluble vitamins are either oxidized or reduced at
DME and can be estimated in a pure solution. Methods have been reported
for following vitamins like riboflavin, nicotinic acid, folic acid, vitamin
B12, vitamin K.

• Estimation of hormones:
• Thyroxine, adrenaline and several hormones give characteristic
polarographs, that under certain conditions can be used for their
determination.
• Determination of alkaloids:
• Most alkaloids gives the reduction steps such as quinine, quinidine, codeine,
morphine etc.

• Miscellaneous:
• Many miscellaneous substances such as sugars, saccharin, antibiotics can be
determined by polarographically. It might include the measurement of
digitoxin in tincture of digitalis, aloin in aloes, soaps and gums etc.

Thermodynamics and thermochemistry

Thermodynamics is the study of the relationship between heat, work, and

other forms of energy.
A thermodynamic system is a body of matter and/or radiation, confined in
space by walls, with defined permeabilities, which separate it from its
surroundings. The surroundings may include other thermodynamic systems,
or physical systems that are not thermodynamic systems.

Thermochemistry is a branch of thermodynamics which is the study of

heat given off or absorbed in a chemical reaction.
Thermal analysis
 Thermal analysis refers to a variety of techniques in which a physical property of a
sample is measured as a function of controlled temperature change.
 Thermal analysis refers to any technique for the study of materials which
involves thermal control.
 Measurements are usually made with increasing temperature, but isothermal
measurements or measurements made with decreasing temperatures are also possible.
Thermal analysis is used

• For characterization of drug substance, excipients, and packaging material:

• For identification of purity, polymorphism, solvation, stability.
• Further the use of these methods for the development of the dosage forms, choice of
the salt form, drug substance—excipient interactions, physical changes on processing or
during storage, and even analysis of the dosage form.
• Thermo-gravimetric analysis
• Principle: TGA measures the amount and the rate of weight change of a material as a function of
temperature or time in controlled environments. Data recorded in form of curve known as
‘Thermogram

• Response ;
– Weight gain – adsorption (physical), oxidation (chemical).
– Weight loss – vaporization (physical), desorption (physical), oxidation (physical),
decomposition (chemical), dehydration & desolvation (chemical).

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 Sample Preparation

Sample preparation has a significant effect in obtaining good data.

It is suggested that maximizing the surface area of the sample in a TGA
pan improves resolution and reproducibility of weight loss temperatures.
The sample weight affects the accuracy of weight loss measurements.
Typically 10-20mg of sample is preferred in most applications.
Whereas, if the sample has volatiles 50-100mg of sample is considered
adequate.
Heating Rate
Samples are heated at a rate of 10 or 20°C/min in most cases. Lowering the
heating rates is known to improve the resolution of overlapping weight
losses.

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• Advances in the technology have made it possible for variable

heating rates (High Resolution TGA) to improve resolution by
automatically reducing the heating rate during periods of weight loss.
• Purge gas; Nitrogen is the most common gas used to purge samples
in TGA due to its inert nature.
• Whereas, helium provides the best baseline.
• Air is known to improve resolution because of a difference in the
oxidative stability of components in the sample.
• Vacuum may be used where the sample contains volatile components,
which helps improve separation from the onset of decomposition
since the volatiles come off at lower temperatures in vacuum. e.g. oil
in a rubber tire product.

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Working
• with help of autosampler sample is placed into a tared TGA sample pan
which is attached to a sensitive microbalance assembly. The thermocouple
sits right above the sample. Care should be taken at all times that the
thermocouple is not in touch with the sample which is in a platinum pan.
• Sample holder is then placed into high temperature furnace. The furnace can
raise the temperature as high as 1000°C which is made of quartz.
• Balance assembly weigh the initial sample at room T & then continuously
monitors changes in sample weight (losses or gains) as heat is applied to
sample.
• Heat applied at certain rate, in various environment. Typical environment:
• ambient air, vacuum, inert gas, oxidizing/reducing gases, corrosive gases,
vapors of liquids or "self-generating atmosphere".
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 Interpretation of TG curves
i. The sample undergoes no decomposition with loss
of volatile products over the temperature range
shown but solid phase transformation, melting ,etc
can not be detected by TG,
ii. The rapid initial mass loss is characteristic of
desorption or drying. If it is true, then re-run the
sample should result in type (i) curves,
iii. Single stage decomposition,
iv. Multi-stage decomposition with relatively stable
intermediates : provide information on the
temperature limit of stability of reactants and
intermediate products and also stoichiometry,
v. Multi-stage decomposition with no stable
intermediate product. However heating-rate effect
must be considered. At low heating rate, type (v)
resemble type (iv). At high heating rate, type (iv)
and (v) resemble type (iii) and lose all the details,
vi. Gain in mass due to reaction with atmosphere,
e.g. oxidation of metals,
vii.Oxidation product decompose again at higher
temperature; this is not often encountered.

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Analytical calculations
Under controlled and reproducible conditions, quantitative data related to mass change also
indicate sample purity or composition.
Example: A pure compound may be either MgO, MgCO3, or MgC2O4. A thermogram of
the substance shows a loss of 91.0 mg from a total of 175.0 mg used for analysis. What is
the formula of the compound? The relevant possible reactions are
MgO  No reaction
MgCO3  MgO+CO2
MgC2O4  MgO+CO2+CO
Solution: % Mass loss Sample=(91.0/175.0)(100%)=52.0
% Mass loss if MgCO3=(44/84.3)(100%)=52.2
% Mass loss if MgC2O4=((44+28)/112.3)(100%)=64.1
If the preparation was pure, the compound present is MgCO3.
 TG curve for CuSO45H2O

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Applications
 Limitations

• Factors affecting TG curve

– heating rate
– sample size
– particle size of sample
– the way it is packed
– crucible shape
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– gas flow rate
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TGA+Spectroscopy/Chromatography
Combination

Gases, vapors
TGA IR or MS or GC

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Differential scanning calorimetry; (DSC) is a thermo-analytical technique in which
the difference in the amount of heat required to increase the temperature of sample and
reference is measured as a function of temperature.
DSC curve

The result of a DSC experiment is a curve of heat flux versus temperature or versus time. This curve can
be used to calculate enthalpies of transitions, which is done by integrating the peak corresponding to a
given transition. The enthalpy of transition can be expressed using equation
Power compensation DSC
https://www.rroij.com/open-access/differential-scanning-calorimetry-a-review-
.php?aid=34700
Applications

Factors effecting DSC

1-Sample nature

• Sample Preparation
• Sample Shape
• Sample Pans
• Sample Weight

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 a-Sample preparation and precautions

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b-Sample Shape
• It is recommended that the sample is as thin as possible and covers as
much of the pan bottom as possible.
• Samples in the form of cakes (as in case of polymers) must preferably
be cut rather than crushed to obtain a thin sample.
• Crushing the sample, whether in crystalline form or a polymer,
induces a stress, which can in turn affect the results.
• In most cases lids should always be used in order to more uniformly
heat the sample and to keep the sample in contact with the bottom of
the pan.
• In case where oxidation properties of a sample are to be studied no lid
is used and the purge gas is usually oxygen to determine Oxidative
Induction Time.

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 c. Sample Pans
• Lightest, flattest pans are known to have the least effect on the results
obtained from a DSC.
• Hermetic pans are used where the sample is expected to have some volatile
content. These pans prevent evaporation.
• Two main reasons for the use of these pans are: The Tg of a polymer or
amorphous material shifts with volatile content.

d. Sample Weight
• Though 5 to 10 mg is considered to be an appropriate sample weight for a
DSC test; A recommendation for metal or chemical melting sample is < 5mg.
• For polymer glass transition Tg or melting sample the mass should be »
10mg.
• Polymer composites or blends the sample mass is >10mg.
• The accuracy of the analytical balance used to measure the sample weight
should be accurate to ± 1%.

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2-Experimental Conditions
a. Start Temperature
• Generally, the baseline should have 2 minutes to completely
stabilize prior to the transition of interest.
• Therefore, at 10°C/min heating rate the run should start at least
20°C below the transition onset temperature.

b. End Temperature
Allowing a 2-minute baseline after the transition of interest is
considered appropriate in order to correctly select integration or
analysis limits.
Care should be taken not to decompose samples in the DSC; it not
only affects the baseline performance but the cell life.
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 c. Reference Pan
• A reference pan of the same type used to prepare the
sample should be used at all times.
• A material in the reference pan that has a transition in
the temperature range of interest should never be
used.
d. Heating Rate
Heating the samples at low heating rates increases
resolution by providing more time at any temperature.
Transitions due to kinetic processes (such as
crystallization) are shifted to lower temperature at
highest cooling rates or higher temperatures at high
heating rates.
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Effects of heating rate

• DSC curves of Acetophenetidin and Phenacetin.
• The Acetophenetidin DSC at 0.5°C/min and
10°C/min showed no effect of heating rate.
• If there were some minor eutectic in this sample then
they would have been detected at the lower heating
rate.
• The melting temperature of pure drugs or chemicals
will have the same extrapolated onset temperature or
the melting point as seen at two varying heating rates.
• The DSC Curve for Phenacetin viewed at heating
rates of 1.0, 5.0 and 20°C/min yielded the same Tm
of 135°C ±1°C.
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 DSC curves of Acetophenetidin

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DSC curves of Phenacetin

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Effect of Heating Rate

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3-Effects of Purge Gases

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Summary of DSC experimental conditions/factors

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Applications Purity by DSC

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 Purity Determination

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How to evaluate melting peaks

• Pure and impure materials:
onset (independent of heating rate)

3
Polymorphic Forms

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Physical Forms of Solids

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Pseudopolymorphism

Amorphous Material

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Study of Polymorphic Transitions.

• Sulfanilamide Polymorphs: It was observed that
sulfanilamide polymorphs are stable and do not show
transition among its forms at heating rates between 1 and
10°C/min.
• DSC of Polymorphs of Tolbutamide: Tolbutamide A
(Form 1) and B (Form 3): When tolbutamide polymorphs
were observed by DSC a significant difference was seen
in their behavior. The difference is due to their structures
which were observed by scanning electron microscope
(SEM).
• The DSC curves are shown below along with the SEM

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 DSC curves of Sulfanilamide Polymorphs

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DSC of Polymorphs of Tolbutamide: Tolbutamide A (Form 1)

and B (Form 3)

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DSC Applications In Pharmaceutical Industry
 Characterization - melting point, heat of fusion,
specific heat capacity, water of crystallization, etc.
 Purity
 Polymorphism
 Screening Tests For Compatibility
 Stability Tests
 Fast and reliable research tool.
 DSC allows fast evaluation of possible incompatibilities,
because it shows change in the appearance, shift or
disappearance of melting, endosperms and exotherms or
variations in the corresponding enthalpies of reaction.
 Rapid analysis, easy handling, high significance for
research, development and quality control.
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Characterization for Pharma

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