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Gas chromatography

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 Gas chromatography
In Gas chromatography, the components of a vapouraised sample are
fractionated as a consequence of a partition between a mobile gaseous phase
and a stationary phase held in a column.

According to the nature of stationary phase, Gas chromatography may be

(a). Gas solid chromatography [GSC]
(b). Gas liquid chromatography [GLC]

Carrier gas
column detector

Basic chromatographic arrangement

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SCHEMATIC DIAGRAM

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PRINCIPLE:

In GLC, the main principle is partition. Gas is used as mobile phase,

liquid which is coated on to a solid support is used as stationary phase. The
mixture of components to be separated is converted to vopour and mixed
with gaseous mobile phase. The component which is more soluble in
stationary phase travels slower and eluted later. The components which is less
soluble in stationary phase travels faster and eluted out first. Not the two
components has the same partition co-efficient for a fixed combination of
stationary phase , mobile phase.

Hence the components are separated according their partition

coefficients.

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THE CHROMATOGRAPHIC PROCESS - PARTITIONING
(gas or liquid)
MOBILE PHASE

Sample Sample
in out

STATIONARY PHASE
(solid or heavy liquid coated onto a solid or support system)

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ADVANTAGES:

 Both qualitative & quantitative analysis are possible.

 Time of analysis is short.

 Instrument is simple & high sensitive.

 the method is applicable to about 60% of the organic compounds.

 Separation, which are very difficult or virtually impossible by other technique,

can be simple & straight forward with GC. The successful separation of pesticides is
a matter of selecting the proper column & conditions.

CRITERIA FOR COMPOUNDS TO BE ANALYSED BY GC:

• Volatility:
• Thermostability:

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Hardware and Columns

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INSTRUMENTATION:
o Mobile Phase: He, Ar, N2, H2

o Flow regulators & flow meters:

o Injection Port: Rubber septum barrier (usually maintained

at a higher temperature than the boiling point of the least
volatile component in the sample mixture)

o Column: (fused silica with a thin coating of stationary phase

on the inner surface)

o Oven: Thermostat controlled forced air oven

o Detector:

o Data System: recorders & integrators

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CARRIER
GAS:
Carrier gas should be:

 Inert, high purity, easily available.

 Low cost, due to large quantities are used

 Allow the detector to respond in an adequate manner

 less risk of explosion or hazards, should give best

column performance consistent with the required speed
of analysis.

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 INJECTION SYSTEM:
Introduced as a plug of vapor with
suitable size
Slow injection or oversized samples
cause band spreading and poor
resolution
Micro syringes
Injection ports

microsyringe
Gastightsyringe 10
 Split/Splitless Injector

Splitless Injection,
(where the split vent is closed)
attempts to transfer all of the
sample to the column and is
used for trace analysis.
Split Mode,
only a small portion
(maybe 1-10% of the sample
moves into the column, and
the rest is sent to waste. This
is used when the analytes are
in high concentration and would
overload the column.
Sample is injected through the
septum with a syringe.
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The oven

Inside here
Column

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 Instrumentation - Oven
Temperature Control
• Isothermal • Gradient

240

200

Temp (deg C)
160

120

80

40

0
0 10 20 30 40 50 60
Time (min)

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Instrumentation - Detectors

Non-Destructive
• Thermal Conductivity (TCD)
• Electron Capture (ECD) [551.1]
• Photo Ionization (PID) [502.2]

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Instrumentation - Detectors
Destructive
• Mass Spectral (CI/EI) [625]
• Flame Ionization (FID) [604]
• Nitrogen-Phosphorus (NPD) [8141A]
• Flame Photometric (FPD) [8141A]
• Electrolytic Conductivity (Hall/ELCD) [502.2]

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 DERIVATISATION:

Precolumn derivatisation: To improve the separation of sample by

Column. (more volatile & thermostable(

Ex: carboxylic acids, sugars, phenols, alcohols, etc can be converted to less polar
compounds by using reagents like BSA reagent(Bis trimetinyl Silyl Acetamide
reagent).
They can also be converted to acetyl derivative or triflouro acetyl derivative.
Post column derivatisation: To improve the response shown by detector . The
components may not be detected by detector unless derivatisation is done.
(online detection technique, where the flow rate is neither stopped nor altered)
The components may be converted in such away that their ionisation or affinity
towards electrons is increased.
Pretreatment of solid support: solid support is to hold the stationary phase
liquid as a thin film.

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 PARAMETERS USED IN GC:
Retention time: Difference in time between the point of
injection and appearance of peak maxima.
Point of injection
Air Peak
peak width Base line
Retention volume: The volume of carrier gas required to
elute 50%of the component from the column.
Vr = retention time*flow rate
Retention time
Peak 1 peak2
Less
separation
Separation factor: The ratio of partition coefficient of the Point of injection factor
two components to be separated. Air
peak Base line

Retention time Rt1

Also watch the following videos: Retention time Rt2

https://www.youtube.com/watch?v=ZpPzImDSfqc More
Peak 1
peak2 separation
https://www.youtube.com/watch?v=08YWhLTjlfo Point of injection
factor

Air
https://www.youtube.com/watch?v=uD-29-mV3N0 peak Base line

Retention time Rt1 17

Retention time Rt2
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