Electrochemistry
 Field of chemistry that focuses on the
interchange between electrical and
chemical energy.
 Use measure potential, current or charge
to determine analyte’s concentration and
characteristics in chemical activity.
 Measurement of the current or voltage
generated by the activity of specific ions.
 Anode (oxidation).
 Cathode (reduction).
Galvanic Cell
Electrolytic Cell
Standard Hydrogen Reference Electrode (SHE)
 This used a universal reference electrode.
 It is typical gas electrode.
Anode – the oxidation reaction occurs.
Cathode – the reduction reaction occurs.
Electricity – flow of electrons over a wire.
Electromotive force - difference of potential
energy of electrons between the two
electrodes.
Oxidation – lose of electrons
Reduction – gain of electrons
Voltaic cell – electrochemical cell that uses
redox reaction to produce else spontaneously.
Salt bridge – maintain electrical neutrally and
allow the reaction to continue.
Cell potential – measure of the tendency of the
cell reaction to proceed toward equilibrium.
Types of electrochemistry
Potentiometry
 Measurement of difference in voltage at
a constant current or under static
conditions.
 Measurement of potential and voltage
between two electrodes in solution by a
null – balance technique.
 The relationship between the nearst
equation.
Application
 Ion selective electrode – ph
electrode – an ISE universally used
in the clinical laboratory
 Parts of ISE
 INDICATOR ELECTRODE
 REFERENCE ELECTRODE
 LIQUID JUNCTION
 READOUT METER
Polarography
 Is the measurement of differences in
current at a constant voltage.
 Used to measure trace metals, oxygen,
vitamin c, and amino acids
concentration
 The relationship between the ilkovic
equation
Coulometry
 Is the measurement of the amount of
electricity in term of coulombs at a fixed
potential.
 Is equal to a current flow of 1 ampere
per second.
 The relationship is expressed by
faraday’s law
Conductometry
 The measurement of the current flow
between two non – polarizable
 electrodes between which is known
electrical potential is establish.
Separation techniques
Chromatography
 Involves the separation of a mixture on
the basis of specific differences of the
physical – chemical characteristics of the
different components on a supporting
medium called absorbent or sorbent.
Kinds of chromatography
 Paper chromatography
- Sorbent: special grade of filter
paper
- Basis of separation: rate diffusion,
solubility of the solute, and nature
of the solvent.
- Use: fractionation of sugars, amino
acids, and barbiturates.
 Gel chromatography
- Other name: gel permeation, size
exclusion, molecular sieve, gel
filtration, and molecular exclusion
- Sorbent: gel with poares of
accurately controlled size.
- Types of gel chromatography:
hydrophilic gels (water – loving) and
hydrophobic gel (water – fearing)
- Basis of separation: molecular
weight and size, charge of the ions,
and hydrophobicity of the
molecules.
 Ion – exchange chromatography
- Sorbent: anion or cation resin with
functional group.
- Basis of separation: differences in
sign and ionic charge densities.
- Use: for the separation of unwanted
substances present in a solution.
 Thin layer chromatography
- It used for drug screening.
- Sorbent: thin plastic plates
impregnated to a layer of silica gel,
alumina, polyacrylamide gel, or
starch gel.
- Basis of separation: rate of
diffusion, solubility of the solute,
and nature of the solvent.
 Liquid – liquid chromatography
- Basis of separation: differences in
solubility between two liquid
phases, aqueous phases, and
organic solvent phase.
- Clinical use: fractionation of
barbiturates and liquid studies.
 Column Chromatography
- Basis of separation: differences in
ph and polarity of solvent.
- Clinical use: fractionation of sugars.
 Gas chromatography
- Types: gas solid chromatography
and gas liquid chromatography
- Basis of separation: sample
volatility, rate of diffusion into liquid
layer, and solubility of sample gas in
the liquid layer.
- Clinical use: drug screening and
drug analysis, and fractionation of
steroids, lipids, barbiturates, blood,
alcohol, and other toxicologic
substances.
 High performance liquid
chromatography
- It follows the concept of selective
adsorption.
- It applies 4,000 to 10,000
Ibs/square inch pressure for the
rapid identification and separation
of high molecular weight
components and many labile
biologic compounds such as
peptides, drugs, hormones,
barbiturates, lipids, steroids, and
antibiotics.
Electrophoresis
 It refers to the migration or movement of
charged particles in an electric field.
 Components: electrical power, support
medium, buffer, sample, and detecting
system.
Amphoteric – is a molecule whose net charge
can be either positive or negative depending on
ph conditions.
Electroendosmosis/ endomosis – movement of
buffer ions and solvent relative to the fixed
support.
Iontophoresis – migration of small charged ions.
Zone electrophoresis – migration of charged
macromolecules.

Factors of rate migration

 Net electric charge
 Size and shape of the molecule
 Electric fluid strength
 Temperature of operation
 Nature of supporting media
Electrophoresis media
 Paper – fragile and easy damaged
staining of protein in variable
 Starch gel – fragile and unable to store
results permanently.
 Cellulose acetate – faster separation
but becomes brittle when drief.
 Agar – basis of electric charge
 Polyacrylamide gel – protein size as
factor.