Introduction to Gas

Chromatography:

Prepared by:
Dr. Mohammed Hamid

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Introduction:
 The father of modern gas chromatography is Nobel
Prize winner John Porter Martin, who also developed
the first liquid-gas chromatograph in (1950).
 Gas chromatography (GC) is a chromatographic
separation technique based on the difference in the
distribution of species between two non-miscible
phases in which the mobile phase is a carrier gas
moving through or passing the stationary phase
contained in a column.
 It is applicable to substances or their derivatives
which are volatilized under the temperatures
employed.
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GC is based on mechanisms of adsorption, mass
distribution or size exclusion.
A sample containing the materials to be separated
is injected into the gas chromatograph. A mobile
phase (carrier gas) moves through a column that
contains a wall coated or granular solid coated
stationary phase. As the carrier gas flows through
the column, the components of the sample come
in contact with the stationary phase. The different
components of the sample have different affinities
for the stationary phase, which results in
differential migration of solutes, thus leading to
separation.

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The sample to be separated should be
thermos-table.
Based on stationary hase used there are
two major types:
A- Gas-solid chromatography:
- Here, the mobile phase is a gas while the
stationary phase is a solid.
- Used for separation of low molecular gases,
e.g., air components, H2 S, CS2 ,CO2 ,rare
gases, CO and oxides of nitrogen.

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B- Gas-liquid chromatography:
- The mobile phase is a gas while the
stationary phase is a liquid retained on the
surface as an inert solid by adsorption or
chemical bonding.

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Principles:
The principle of separation in GC is
adsorption or “partition.”
The mixture of component to be separated
is converted to vapour and mixed with inert
gaseous mobile phase.
The component which is more soluble in
stationary phase travel slower and eluted
later. The component which is less soluble
in stationary phase travels faster and eluted
out first.
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No two components has same partition
coefficient conditions. So the components
are separated according to their partition
coefficient.
Partition coefficient is “the ratio of
solubility of a substance distributed
between two immiscible liquids at a
constant temperature.’

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Instrumentation:
The instrument consists of carrier gas, an
injector (sample injection system), a
chromatographic column contained in an
oven, a detector and a data acquisition
system (or an integrator or a chart
recorder).
1- Carrier gas:
Usually the carrier gas is contained in
metal cylinder.

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The cylinder/ gas tank is fitted with a
pressure controller to control the pressure
of gas, a pressure gauge that indicates the
pressure, a molecular sieve to transfer
filtered dry gas and a flow regulator to
ensure a constant rate of flow of mobile
phase to the column.
Factors are considered while selecting a
carrier gas:
a) Should be chemically inert.
b) Should be cheap and readily available.
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c) Should be of high quality and purity and
not cause any fire accidents.
d) Should give best possible results.
e) Should be suitable for the sample to be
analyzed and for the detector.
f) Free from moisture.
g) Safe in handling.

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Flow rate required for most detectors is
30 -40 ml/min.
Hydrogen, helium, nitrogen and carbon
dioxide are commonly used.
Hydrogen has low density and better
thermal conductivity. However, it reacts
with unsaturated compounds and is
inflammable and explosive in nature.
Nitrogen is inexpensive but it gives
reduced sensitivity.

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Helium is the most preferred gas.
Inlet pressures usually range from 10 to
50 psi greater than the room pressure,
which lead to flow rates of 25 to 150
ml/min for packed column and 1 to 25
ml/min with open-tubular capillary
columns.

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2- Sample injection system:
Sampling unit or injection port is attached
to the column head.
Since the sample should be in vapourized
state, the injection port is provided with
an oven that helps to maintain its
temperature at about 20-500° C above the
boiling point of the sample.

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Gaseous samples may be introduced by
use of a gas tight hypodermic needle of
0.5-10 ml capacity.
For Liquid samples , micro syringes of
0.1-100μL capacity may be used.
Column efficiency requires that sample
be of suitable size & be introduced as a
plug of vapour.
Slow injection or over sized samples
cause band spreading & poor resolution.

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3- Column unit:
Two general types of columns are encountered
in GC:
a) Packed columns
b) Capillary columns or open tubular columns.
Packed column length 1.5 -10 m and Capillary
column 10-100 m.
Constructed of stainless steel, glass, fused silica or
Teflon.
To fit into an oven for thermostating, they are
usually formed into coils having diameters of 10
to 30 cms.
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a) Packed columns:
 Have inside diameter of about 2 to 4mms.
 Length- 1.5 - 10m.
 These tubes are densely packed with a uniform, finely divided
packing material or solid support (diatomaceous earth, glass beads),
that is then coated with a thin layer of stationary liquid phase.
 The packed columns are equilibrated before introduction of the
sample. This is done by allowing continuous flow of heated carrier
gas through the columns for a specific duration of time (24hrs) at
prescribed temperature.
 Ideally prepared and conditioned columns show a zero base line on
the recorder upon passage of carrier gas alone.
 Solid support materials:
 Purpose- It serves to hold the liquid stationary phase in place so
that as large a surface area as possible is exposed to the mobile
phase. E.g –naturally occurring diatomaceous earth, glass beads.

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 Ideal characters:
- Should be small, uniform, spherical.
- Good mechanical strength.
- Specific surface area of at least 1m2/g.
- Inert and stable at elevated temperatures.
- Uniform wetability.
 Support materials are often treated chemically
with dimethylchlorosilane.
 The HETP is proportional to average particle
diameter so that smallest possible particles should
be preferred in terms of column efficiency.
 Particle size is 60 to 100 mesh (250 to 170 um).

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b) Open tubular or capillary columns:
length ranges from 10-100m.
inner diameter is usually 0.1-0.5mm.
It is mainly of two types:
- Wall-coated open tubular columns (WCOT).
- Support -coated open tubular columns (SCOT).
- Wall-coated open tubular columns (WCOT):
- They are simply capillary tubes coated with a
thin layer of the stationary phase.
- WCOT columns are constructed with stainless
steel, aluminum, copper, plastic & glass.

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The column is treated with gaseous HCl,
strong aqueous HCl, or potassium hydrogen
fluoride to give a rough surface, which bonds
the stationary phase more tightly.
- Support -coated open tubular columns
(SCOT):
- the inner wall of the capillary is lined with a
thin layer of support material such as
diatomaceous earth, onto which the stationary
phase has been adsorbed. It is also known as
PLOT (porous-layer open tubular columns).

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-SCOT columns are generally less efficient
than WCOT columns but have a higher
sample capacity which enable them to be
used without a sample splitter. Both types
of capillary column are more efficient than
packed columns.
A recent advancement in capillary columns
are fused-silica open tubular columns
(FSOT) columns. They are drawn from
specially purified silica that contains
minimal amounts of metal oxides.
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LIQUID STATIONARY PHASES:
It should have the following properties:
a) It should be non-volatile.
b) Should have high decomposition
temperature (thermally stable).
c) Should be chemically inert.
d) Should posses low vapour pressure at
column temperature.
e) Should be chemically and structurally
similar to that of the solute i.e., polar for
polar solute.
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The proper choice of stationary phase is
often crucial to the success of a separation.
Some important facts about stationary
phase:
The retention time for an analyte on a
column depends on its distribution constant,
which in turn is related to the chemical
nature of the liquid stationary phase.
To separate various sample components, their
distribution constants must be sufficiently
different to accomplish a clean separation.
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At the same time, these constants must not be
extremely large or extremely small.
To have a reasonable residence time in the
column, an analyte must show some degree of
compatibility (stability) with the stationary
phase.
Principle involved “Like dissolves Like”.
Generally the polarity of the stationary phase
should match with that of the sample
components. When the match is good, the order
of elution is determined is determined by the
boiling point of eluents.
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Some common liquidstationary phases used
in GLC:
STATIONARY TRADE MAX. APPLICATIONS
PHASE NAME TEMP, °C

Polydimethyl OV-1, 350 General purpose

siloxane SE-30 nonpolar phase,
hydrocarbons,
polynuclear aromatics,
steroids.
5% Phenyl - OV-3, 350 Fatty acid methyl esters,
Polydimethyl Se-52 alkaloids, drugs,
siloxane halogenated compounds.
50% Polydimethyl OV-17 250 Drugs, steroids,
phenyl -siloxane pesticides, glycols.

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STATIONARY TRADE MAX. APPLICATIONS
PHASE NAME TEMP, °C
50% OV-210 200 Chlorinated
Trifluoroproply - aromatics, nitro
Polydimethyl aromatics, alkyl
siloxane substituted
benzenes
Polyethylene Carbowax 20M 250 Free acids,
glycol alcohols, ethers,
essential oils,
glycols.
50% Cyanopropyl OV-275 240 Polyunsaturated
-Polydimethyl fatty acids, rosin
siloxane acids, free acids,
alcohols.

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Examples of different liquid phases
categories:

CATEGORY EXAMPLES

Non-polar hydrocarbon phases Paraffin oil (nujol), silicon oil,

silicon rubber gum (used for
high temp of about 400°
Polar compounds (having polar Polyglycols (carbowaxes)
groups like -CN, -CO and –OH)
Liquids having hydrogen Glycol, glycerol, hydroxy acids
bonding

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4- Detector:
The eluted solute particles along with the
carrier gas exit from the column and enter
the detector.
The detector then produces electrical
signals proportional to the concentration
of the components of solute.
The signals are amplified and recorded as
peaks at intervals on the chromatograph.

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Properties of an ideal detector:
- Sensitive.
- Operate at high T (0-400 °C).
- Stable and reproducible.
- Linear response.
- Wide dynamic range.
- Fast response.
- Simple (reliable).
- Nondestructive.
- Uniform response to all analytes.
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a) Thermal conductivity detector (TCD):
Also called as Katharometer or Hot wire
detector.
Principle:
“TCD is based upon the fact that the heat
lost from a filament depends upon the
thermal conductivity of the stream of
surrounding gas as well as its specific
heat.”
Thus the rate of loss of heat is related to the
composition of the surrounding gas.
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TCD is non-destructive, concentration sensing
detector. A heated filament is cooled by the
flow of carrier gas.
When the carrier gas is contaminated by
sample, the cooling effect of the gas changes.
The difference in cooling is used to generate
the detector signal.
TCD filaments are made up of fine platinum,
gold or tungsten wire.
TCD thermistors are made of Oxides of
manganese, Cobalt or nickel to which some
trace elements are added.
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Advantages:
- Simple and inexpensive.
- Durable and posses long life.
- Large linear dynamic range.
- Accurate results.
- Non-selective, hence known as universal detectors.
- Non destructive character, which permits collection
of solutes after detection.
Disadvantages:
- Low sensitivity.
- Affected by fluctuations in temperature and flow
rate.
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b) Electron capture detector:
It responds to only those compounds whose
molecules have an affinity for electrons.
E.g. : halogen containing organic compounds
such as pesticides & polychlorinated
biphenyls; peroxides; quinones & nitro group
containing compounds.
On the other side, it responds very little to
compounds such as hydrocarbons, alcohols,
amines.

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 Principle:
 Electron capture detector is an ionization detector and
its response is based upon the ability of molecules with
certain functional groups to capture electrons generated
by the radioactive source (e.g. Ni63). The detector
chamber contains two electrodes (cathode and an
anode), and a radioactive foil as the radiation source.
Using an inert carrier gas with no affinity for electrons,
the ions formed by the ionizing radiation can be
collected, creating a steady standing current in the
detector. When molecules of certain electron-absorbing
solutes enter the detector chamber, they will capture
electrons, resulting in a decrease of the standing
current, giving a peak.

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Advantages:
- Highly selective.
- Highly sensitive for the detection of
compounds like halogens, quinones,
peroxides, nitrites, etc.
- It is non-destructive.
- More sensitive than TCD and FID.
Disadvantage:
- Least sensitive to compounds whose
molecules have negligible affinity for
electrons.
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c) Flame ionization detector:
 The flame ionization detector (FID) is the most
commonly used detector in GC.
 This employs hydrogen flame that is maintained in a
small cylindrical jet made up of platinum or quartz.
 Effluent from the column with helium or nitrogen as
carrier gas are fed into the hydrogen flame, gets
ignited and undergoes pyrolysis to produce ions.
 When collected, the ions produce a small current
that provides the desired signal. When no sample is
being burned, there should be minimal ionization
and thus the blank signal is very small.

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Other detectors are summarized in the
table below:

Detector Applications
Photoionization (PID) Aromatic compounds
Nitrogen/phosphorus (NPD) N-, P-, and halogen-containing
compounds
Flame photometric (FPD) S- and P-containing compounds
Atomic emission (AED) Metals; halogens, C- and O-
containing compounds
Electroconductivity (ECD) S-, N-, and halogen-containing
compounds
Chemiluminescent S-containing compounds
Radioactivity H3 - and C14
-containing compounds
Mass spectrometer (MSD) Variety of compounds

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 Practical Working steps:
- Preparation of sample according to method of analysis.
- Fill the syringe with sample.
- Record the setting i.e., column temperature, detector
temperature and injection port temperature.
- Introduce sample into the injection port by completely inserting
the needle into the rubber septum. Note down the injection time.
- The sample gets vapourized due to higher temperature of
injection port and is swept into column by carrier gas.
- This sample components now get distributed between the gas
and stationary phase depending upon their solubilizing
tendencies.
- The components with minimal solubility move faster and those
with maximum solubility travel slowly.
- The components leaving the column activate detector and
recorder to give a plot.
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Tha
nk
you
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