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Gas Chromatography
Gas Chromatography
Gas Chromatography
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• Gas Chromatography (GC) is a column
chromatographic technique in which the mobile
phase is a gas and stationary phase is a liquid or
solid and the sample components are separated as
vapours.
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• Gas chromatography has a potential role both in
the qualitative and quantitative analysis.
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• It is thus used to separate and detect small
molecular weight compounds in the gas phase.
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Types of GC
• Gas-liquid chromatography (Stationary phase
is liquid)
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Gas-liquid chromatography
• In this type, the stationary phase is a thin layer
of non-volatile liquid bound to an inert solid
support
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Gas-solid chromatography
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Principle
• During a GC separation, the sample is vaporized and
carried by the mobile phase (the carrier gas) through the
column. Separation of the different components is
achieved based on their relative vapor pressure and
affinities for the stationary phase. The affinity of a
substance towards the stationary phase depends upon
distribution constant Kc, or partition coefficient of sample
between stationary phase and mobile phase
Kc = Cs/Cm,
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Instrumentation
• Carrier gas (maintained at high pressure)
• Flow regulator
• Column
• Detectors
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Schematic Diagram of Gas
Chromatograph
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Carrier gas (Mobile Phase)
• It should be chemically inert
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Carrier gas (Mobile Phase)
• The role of the carrier gas or mobile phase in GC is to
carry the sample molecules along the column while
they are not dissolved in the stationary phase.
• The carrier gas is inert and does not interact with the
sample.
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Sample injector
• The carrier gas also enters the chamber.
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Injecting syringe Insertion point
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Sample injection system
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Column
• There are two general types of column used in GC -
packed column and capillary column (open tubular).
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• Capillary columns have an internal diameter of 0.1-0.5
mm. They can be one of two types; wall-coated open
tubular (WCOT) or support-coated open tubular (SCOT)
column. Wall-coated columns consist of a capillary tube
whose walls are coated with liquid stationary phase.
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• SCOT columns are generally less efficient than WCOT
columns. Both types of capillary column are more
efficient than packed columns.
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GC columns
glass stainless steel copper
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Thermostatic Chamber or Oven
• For precise work, column temperature must be
controlled to 1/10th of a degree.
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Inside GC oven
HIMALAYAN PHARMACY 30
INSTITUTE
Detectors
• Flame ionization detector
• Mass detectors
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Thermal Conductivity Detector
• Katharometer or hot wire detector
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Thermal Conductivity Detector
• Both the filaments are connected to the arms of
wheatstone bridge arrangement.
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Flame Ionisation Detector
• FID is the most widely used and generally applicable
detector
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Flame Ionisation Detector
• The resulting current is then measured which is
directly related with the concentration of eluent eluted
from the column.
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Electron Capture Detector
• Is used for detecting electron-absorbing components
having high electronegativity such as halogenated
compounds and nitro compounds
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Electron Capture Detector
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Derivatization
• It has been estimated that 10-20% of known
compounds can be directly analysed by GC
• Alkylation
• Acylation
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Alkylation
• A reactive hydrogen such as –OH or -NH is replaced by
alkyl group to reduce the molecular polarity.
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Silylation
• It is the most versatile and widely used derivatisation
technique in GC
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Acylation
• Acylation with acid anhydrides or acyl imidazoles
converts primary and secondary amines to stable
derivatives.
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Advantages
• Due to its high efficiency, GC allows the separation of
the components of complex mixtures in a reasonable
time.
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Disadvantages
• Limited to thermally stable and volatile compounds.
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Applications
• Measurment of drugs and other metabolites in
biological fluid
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Applications
•Determine the sub-micro drug level in biological
specimen like blood, urine.
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