GAS CHROMATOGRAPHY

 GC is useful for compounds which are

naturally volatile or can be converted into
a volatile form.

 GC has been a widely used method due to

its high resolution, low detection limits,
accuracy and high resolution limits, and
short analytical time.

 Retention of a compound is determined by

its vapour pressure and volatility which, in
turn, depends on its interaction with
stationary phase.
 Two types of stationary phases are
commonly used in GC are solid absorbent
(Solid Gas Chromatography [GSD]), and
liquid coated on solid support (Gas-Liquid
Chromatography [GLC]).
 In GSC same material (usually silica,
alumina, or activated carbon) act as both
the stationary phase and support phase.
 GLC uses liquid phases such as polymers,
hydrocarbons, fluorocarbons, liquid
crystals, and molten organic salts to coat
the solid support materials.
 Calcin diatomaceous earth graded into
appropriate size ranges used as a
stationary phase because it is stable
inorganic substance.
 Components of a typical GC system
consists five components: a gas
cylinder as a mobile phase source, a
sample injector, a column, a detector,
and a computer for data acquisition.
 The mobile phase used in GC usually
inert gas such as nitrogen, helium,
hydrogen, argon, carbon dioxide,
nitrous oxide and ammonia.
 The carrier gas should be of high
purity, and the flow must be tightly
controlled to ensure optimum column
efficiency and reproducibility of test
results.
 Samples are introduced in to GC using
hypodermic syringe or automated
samplers.
 Each injection port is heated to very
high temperatures to derivatize it into
more volatile form by derivatization
reactions like silylation, alkylation, and
acylation.
• Silylation is most common technique
which replaces active hydrogen on
compounds with alkylsilyl group. This
substitution results in a more volatile
form that is less polar and more
thermally stable.
• Retention of compounds in a GC
column can be adjusted by changing
temperature.
• The column temperature affects
volatility of compounds and thus the
degree of their interaction with the
stationary phase.
High performance Liquid
Chromatography
 HPLC uses small rigid support and special
mechanical pumps producing high pressure
to pass the mobile phase through the column.
 HPLC column may be used several times
without regeneration.
 The resolution achieved by HPLC is superior
than any other forms of liquid
chromatography.
 There are five commonly used separation
techniques in liquid chromatography.
 They include adsorption, partition, ion
exchange, affinity and size exclusion.
 Each is characterized by a unique
combination of stationary phase and mobile
phase.
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