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CHROMATOGRAPHY
(GC)
SAIMA ALEEM.
M.PHIL.
Course no CHM -754.
COURSE SUPERVISOR: DR RAHAT
GAS CHROMATOGRAPHY
INTRODUCTION
Gas chromatography is one of the most versatile and ubiquitous
analytical techniques in the laboratory. It is widely used for the
determination of organic compounds.
Complex mixtures can be separate by this technique.
A gas chromatography is a chemical analysis instrument for separating
chemicals in a complex sample.
A gas chromatograph uses a flow-through narrow tube known as
the column, through which different chemical constituents of a sample
pass in a gas stream (carrier gas, mobile phase) at different rates.
It depending on their various chemical and physical properties and their
interaction with a specific column filling called the stationary phase.
As the chemicals exit the end of the column, they are detected and
identified electronically. The function of the stationary phase in the
column is to separate different components, causing each one to exit the
column at a different time (retention time).
Other parameters that can be used to alter the order or time of
retention are the carrier gas flow rate, column length and the
temperature.
Gas chromatography
In a GC analysis, a known volume of gaseous or liquid analyte is injected
into the "entrance" (head) of the column, usually using a
microsyringe (or, solid phase microextraction fibers, or a gas source
switching system).
As the carrier gas sweeps the analyte molecules through the column,
this motion is inhibited by the adsorption of the
analyte molecules either onto the column walls or onto packing
materials in the column.
The rate at which the molecules progress along the column depends on
the strength of adsorption, which in turn depends on the type of
molecule and on the stationary phase materials.
Since each type of molecule has a different rate of progression, the
various components of the analyte mixture are separated as they
progress along the column and reach the end of the column at different
times (retention time). A detector is used to monitor the outlet stream from the
column.
The time at which each component reaches the outlet and the amount of that
component can be determined.
AUTO SAMPLER
The auto sampler provides the means to introduce a sample automatically into the
inlets.
Liquid.
Static head-space by syringe technology.
Dynamic head-space by transfer-line technology.
Solid phase micro extraction (SPME).
CARRIER GAS
By using a syringe with a long needle, the tip can be made to penetrate
past the liner and discharge its contents directly into the column
packing.
This procedure is called 'on-column injection' and, as it reduces peak
dispersion on injection and thus, provides higher column efficiencies, is
often the preferred procedure.
Open Tubular Column Injection Systems
Due to the very small sample size that must be placed on narrow bore
capillary columns; a split injection system is necessary.
As the sample passes the column opening, a small fraction is split off
and flows directly into the capillary column are called a split injector.
Regulating the portion of the carrier gas that passes to waste changes
the split ratio. This achieved by an adjustable flow resistance situated in
the waste flow line. This device is only used for small diameter capillary
columns where the charge size is critical.
The device has certain disadvantages due to component differentiation
and the sample placed on the column may not be truly representative.
The solutes with the higher diffusivities (low molecular weight) are lost
preferentially to those with lower diffusivities (higher molecular
weights). Consequently, quantitative analyses carried out using the high
efficiency small diameter capillary columns may have limited accuracy
and precision, depending on the nature of the sample.
Packed columns
Qualities of the packed column
Glass column must be used if any of the sample components are
decompose by compact with metal.
The material chosen as a inaet support should be uniform granular size
and capable of being packed in to a uniform bed in a column.
The surface area of the material should be large so as to promote
distribution of the liquid phase as a film and ensure the rapid
attainment of equilibrium between the stationary phase and mobile
phase.
exceeding 20% without becoming too sticky to flow freely and can be
easily packed.
To be a good resolution the height is equivalent of the theoretical plate
is proportion to the average particle diameter.
Rapidly increase the pressure due to decrease the particle size.
It is necessary to increase the pressure for achieving the flow rate
through the column
Open columns
These capillary columns (i.d < 1mm) are increasingly used in GLC.
It is used for complex mixtures.
This result from a highly theoretical plate numbers which can be attain
with long column of this type for the relatively smaller pressure drop.
In these capillary columns the stationary phase s coated on the inner
wall of the tube,
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Supported coated open tubular is less efficient then wall coated open
tubular.
1. SENSITIVITY
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The linear range of the detector refer to the concentration range over
which the signal is directly proportional to the concentration of analyte.
Linearity in detector response will give linearity of the calibration graph
and allow later to be drawn with more certainty.
With the convex calibration curved, the precision is reduced at higher
concentrations where the slope of the curved is much lesser.
The high linearity range has high advantage but detector small range
will be used because of their other qualities, although they will need to
be recalibrated over a number of difference concentration ranges.
3. Stability
The important characteristic of a detector is the extent to which the
signal output remains constant with time, assuming there is a constant
input.
Lack of the stability can be exhibited in to two ways.
1. Baseline noise.
2. Drift.
1 .Baseline noise.
Baseline noise is caused by a rapid random variation in detector output
difficult to measure small peaks against the fluctuation background.
2 .Drift
Drift is often due to factors external to the detector and impose a more
fundamental limit on its performance, such as temperature change or
column bleed and so is controllable, where as noise is usually due to
poor contacts within detector and impose a more fundamental limit on
its performance.
It must be reproducible
TYPE OF DETECTOR
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Helium and hydrogen are the best carrier gases to use in conjunction
with this type of detector. Since their thermal conductivity are much
higher than the other gases.
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When the pure carrier gas is pass over both the Sample and reference
filaments the bridge is balanced but when the vapours emerges from
the column, the rate of cooling of the sample filament become changed
and the bridge become unbalanced.
It is used for the detection limit of permanent gases light hydrocarbon
and compound which response poorly from the flame ionization
detector.
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TYPES OF CHROMATOGRAPHY
Gas solid (adsorption) chromatography.
Gas liquid (partition) chromatography.
The most important one of the two is gas liquid chromatography, used
in the foam of capillary column.
GAS-LIQUID CHROMATOGRAPHY
In gas-liquid chromatography, the mobile phase is a gas such as helium
and the stationary phase is a high boiling point liquid absorbed onto a
solid.
chromatography
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