GAS

CHROMATOGRAPH
Y

P R ES EN TED BY: 1 . S AM PA
M ON D AL
2 . PAY EL TR I PATH I
 CONTENTS
 INTRODUCTION OF
CHROMATOGRAPHY
 CLASSIFICATION OF
CHROMATOGRAPHY
 GAS CHROMATOGRAPHY
 TYPES OF GAS
CHROMATOGRAPHY

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 CHROMATOGRA
PHY
 Chromatography is an important biophysical technique that enables the
separation, identification, and purification of the components of a mixture for
qualitative and quantitative analysis.
CLASSIFICATION OF CHROMATOGRAPHY:
CHROMATOGHAPHY

Stationary Mobile Based on force of

Phase separation
• Thin Phase
Layer • Liquid • Adsorption
Chromatography Chromatography Chromatography
• Paper • Gas • Partition
Chromatography Chromatography Chromatography
• Column • Ion exchange
Chromatography Chromatography
• Gel filtration
Chromatography
• Affinity 3
 GAS CHROMATOGRAPHY
 It is a process of separating component(s) from the given crude drug
by using a gaseous mobile phase.
 This chromatographic technique can be used to separate volatile
organic compounds.
TYPES OF GAS CHROMATOGRAPHY: There are two types of
chromatography as follows-
GAS-LIQUID • In this technique, the liquid phase which is
coated on a solid support of a capillary tube.
CHROMATOGRAPH • Principle involved in Gas-liquid chromatography
Y is Partition.

GAS-SOLID • In this technique, the stationary phase is solid

CHROMATOGRAPH adsorbent.
• Principle involved in Gas-solid chromatography
Y is Adsorption.

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PRINCIPLE:
 The principle of Gas Chromatography (GC) is Partition.
 The mixture of component to be separated is converted to vapour & mixed
with gaseous mobile phase.
 The component which is more soluble in stationary phase travel slower &
eluted later. The component which is less soluble in stationary phase travel
faster & eluted out faster.

STATIONARY AND MOBILE PHASES IN GAS

CHROMATOGRAPHY:
 Stationary Phase- Non volatile liquid coated on inner support.
 Mobile Phase- Hydrogen, nitrogen, helium, carbon dioxide and argon
are some example of carrier gas (mobile phase) used in GLC.

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CRITERIA FOR COMPOUNDS TO BE ANALYSED BY GAS
CHROMATOGRAPHY:
Two important criteria for analysed by Gas Chromatography are-
 Volatility: Unless a compound is volatile, it cannot be mixed with mobile phase.
Hence volatility is important.
 Thermostability: All the compounds will noy be in the form of valour. There will
be solid as well as liquid samples. Hence to convert them to a vapour form, they
have to be heated to a higher temperature. At that temperature the compound
have to be thermostable. If they are not thermostable, the compounds can’t be
analysed by gas chromatography since they will be decomposed.

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INSTRUMENTATION:

 Carrier gas
 Flow regulators and flow meters
 Injection devices
 Columns
 Temperature Control Devices
 Detectors
 Recorders and Integrators

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Fig: Gas Chromatography

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1. Carrier Gas:
• Hydrogen- Good thermal conductivity, low density,
inflammable.
• Helium- Good thermal conductivity but expensive.
• Nitrogen- Inexpensive, reduced sensitivity.
2. Flow regulators & Flow meters:
• Rotameter-
I. Placed before the column inlet.
II. It has glass tube with a float held on spring.
III. Level of float is determined by flow rate of
carrier gas.
Fig:
Rotameter

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• Soap Bubble Meter- consist of soap solution in
rubber bulb.
Gas
enters the
meter

Bulb will
be gently
pressed

Drop of soap
solution converted
to bubble by
pressure of carrier
gas Fig: Soap Bubble
Distance
travelled by Meter
the bubble
indicates flow
rate

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3. Injection Devices:
• Direct injection is mostly used.
• There are classified into four types-
I. Automatic sampler
II. Headspace sampler
III. Purge & Trap
IV. Pyrolysis
4. Columns:
• Most important part of the GC.
• May be made up of Glass or Stainless
steel.
• It is mainly two types-
I. Depending on Use
II. Depending on Nature

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Depending on Depending on
Use Nature Wall
coated
Analytical Capillary
Column Column
Support
Preparative Packed coated

Column Column

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5. Temperature control Devices:
Two operations are available-
• Isothermal Programming same temperature throughout process.
• Linear Programming oven heated linearly the period of time.
6. Detectors:
A. THERMAL CONDUCTIVITY DETECTOR
B. FLAME IONISATION DETECTOR
C. THERMIONIC DETECTOR
D. ELECTRON CAPTURE DETECTOR

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 THERMAL
CONDUCTIVITY
DETECTOR

 The principle is based upon thermal conductivity difference between carrier

gas and that of component.
 It is simple, easy to maintain & inexpensive.
 In this detector, the sample is not destroyed & hence used in preparative
scale.
 The sensitivity of this detector is low & cannot be analysed biological samples.
 The thermal conductivities of some carrier gases are as follows:
HYDROGEN HELIUM NITROGEN METHANE HEXANE

32.7 33.9 5.2 6.5 3.0

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 FLAME
IONISATION
DETECTOR

 The ionisation detectors are based upon the electrical conductivity of

carrier gas.
 At normal temperature & pressure, gases act as insulators, but
become conductive if ions are present.
 This detector is stable and insensitive to small changes in the flow
rate of carrier gas and water vapour.

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 THERMIONIC
DETECTOR

 Also known as alkali flame ionization detector.

 This is used for detecting nitrogen or phosphorus containing
compounds.

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 ELECTRON
CAPTURE
DETECTOR

 This detector is radiation-based detector.

 This is mainly selective for compounds containing electronegative
atoms, such as halogens.

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 PARAMETERS USED IN GAS
CHROMATOGRAPHY
1. Retention time (Rt):
• Retention time is the difference in time between the point of injection &
appearance of peak maxima.
• Retention time is the time required for 50% of a component to be eluted from a
column.
• This time is measured in minutes or seconds.
2. Retention volume (Vr):
• It is the volume of carrier gas required to elute 50% of the component from the
column.
• It is the product of retention time & flow rate.
Retention volume= Retention time * flow rate
3. Separation factor:
• It is the ratio of partition coefficient of the two components to be separated.

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4. Resolution:
• It is a measure of the extent of separation of two
components & the baseline separation achieved.
• It can be determined by using the following formula-
Rs=
5. Theoretical Plate:
• It is an imaginary or hypothetical unit of a column where
distribution of solute between stationary phase & mobile
phase has attained equilibrium.
• A theoretical plate can also be called as a functional unit of A= Eddy diffusion
term or multiple
the column. path diffusion
6. Height equivalent to a Theoretical Plate (HETP): B= Longitudinal
• It can be of any height, which decides the efficiency of diffusion term
separation. C= Effect of mass
• If HETP is less, the column is more efficient. transfer which
• HETP can be calculated by using the following formula- depends on flow rate
HETP= U= Flow rate or
[HETP is given by the Van Deemeter equation] velocity of the
mobile phase
HETP=A++Cu
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APPLICATIONS:
1. Qualitative analysis- It is nothing but identification of a compound.
This is done by comparing the retention time of the sample as well as
the standard. Under identical conditions, the retention time of the
standard & the sample are same. If there is a deviation, then they are
not the same compound.
2. By comparing the chromatogram of the standard & that of the sample,
the purity of the compound can be reported.
3. The percentage of impurities may also be calculated from peak areas.

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