Gas chromatography
1
Chromatograms -
Supelco® PTE-5 Supelco
2
Gas chromatography involves separation
and analyses of different constituents of
mixtures by a mobile gas phase passing
over a stationary adsorbent. The technique
is similar to column chromatography
except that the mobile phase is replaced by
a moving gas which is called the carrier
gas.
Gas chromatography is a powerful tool for
the analyses of organic materials.
3
Gas chromatography can be of two types:
Gas-liquid Chromatography (GLC) and
Gas-Solid Chromatography.
In gas liquid chromatography the
separation is brought about by partitioning
the sample between a mobile gas phase
and a thin nonvolatile liquid layer coated
on some inert solid particles while gas-
solid chromatography is based upon
selective adsorption of constituents of the
sample on a solid of large surface area
used as the stationary phase.
When a mixture of volatile material
transported by a carrier gas is led through
a column containing an adsorbent solid
phase or more commonly an absorbing
liquid phase coated over a solid material,
each volatile component is partitioned
between the carrier gas and the solid or the
liquid.
Depending upon the retention time in the
column, the volatile components emerge
from the column at different times and are
finally detected by a suitable detector. If
4
the carrier gas used, the rate of flow, and
the temperature of the column is kept
constant the retention time for each for
each constituent of the mixture will always
be the same.
Packed column ~ o.5 cm i.d , 2-5 m L
Capillary column: 200 micrometr i.d and
10-30 m long
5
Retention time shows a linear relationship
with the boiling point of the compound
and is characteristic for the constituents
concerned under the given set of
conditions for a given column. Thus, it is
possible to identify the compound from
its characteristic retention time on a
particular column and under a given set of
conditions.
Quantitative estimation can be carried out
from the extent of peak area recorded by
the detector recorder system.
Apparatus used for gas chromatography is
a simple tube of about 4 mm in diameter
and about 120 cm to many metres in
length. It is made of stainless steel or glass
and is usually bent or coiled so that it
could be accommodated in a small space.
The tube or the column is packed with
particles of some suitable adsorbent or in
the case of gas-liquid chromatography the
fixed phase is a non-volatile liquid coated
over some solid support (particles of
6
diatomaceous earth, crushed fire bricks
etc.).
As mobile phase, the gases used may be
Argon, Helium, Nitrogen or Hydrogen -
Hydrogen is usually not preferred because
of fire hazards. There has been a recent
trend to use capillary-gas chromatography
in which instead of the column a very thin
capillary, about 0.25 mm in diameter and
usually 50 metres in length made of glass,
stainless steel or some organic polymer is
employed. The inside of the capillary wall
is coated with the stationary liquid phase.
These capillary columns are superior to
packed columns in terms of separation
efficiency. They can separate upto several
components from a single sample.
In order to maintain a constant rate of flow
of the carrier gas, there is a flow meter and
an adjustment device which regulates the
flow of the carrier gas into the column.
The sample is introduced through a self
sealing silicon rubber partition into a
chamber which is heated to bring about
7
evaporation of the sample. The
temperature of the chamber must not be so
high as to decompose the sample.
Solid samples have to be dissolved in
some solvent whereas gaseous samples
require special sample introduction valves.
The detectors, placed at the exit of
separation chamber, detect and measure
the small amounts of separated
components present in the stream of the
carrier gas leaving the column. Normally
three types of detectors are employed in
gas chromatography:
1- thermal conductivity detectors,
2- flame ionization detectors
3- electron capture detectors.
Thermal conductivity detectors are the
most widely used detectors in gas
chromatography. These detectors use
heated metal filament (or thermisters
which are made of some semi-conductor
of fused metal oxides) to sense small
8
changes in thermal conductivity of the
carrier gas. Thermal conductivity of the
carrier gas only gives an essentially
constant signal. The presence of vapours
of the different components of the mixture
in carrier gas brings about changes in the
thermal conductivity proportional to their
amount in the stream. This brings about
changes in the resistance of the filament
which is measured. The recorder which
records these changes is equipped with an
automatic device which traces the
magnitude of these changes on a graph
sheet along with the retention time.
Flame ionization detectors are based on
the measurement of electrical conductivity
of gases. At normal temperatures and
pressure, gases act as a bad conductor or
insulators but when ionized they act as a
good conductor of electrical current.
Gases and vapours as they emerge from
the separation column are mixed with
hydrogen and burned in air to produce a
9
flame which ionizes the component
molecules in the carrier gas. The burning
jet is the negative electrode while the
anode is usually a small loop of wire
extending into the tip of the flame across
which a small voltage is applied.
The ions produced are collected at the
electrodes and a current is generated which
is proportional to the number of the
component molecules ionized. Only the
carrier gas burning with hydrogen
produces an essentially constant signal but
when the components of the mixture being
analysed emerge ionization occurs and a
higher current is observed. The detector is
equipped with an automatic recording
device which records these fluctuations
and transmits them to a graph sheet along
with the retention time.
10