University of Bahrain

College of Science
Department of Chemistry

Experiment 6
Gas Liquid Chromatography

Date the Experiment was Done: 2010-4-1

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 Index

Page
o Objective……………..………………….3
o Abstract…………………………………..3
o Theory……...…………………………….3
o Requirements……………………………3
o Results……………………….…………..5
o Conclusions and Recommendations.....6
o References…………………………….….6

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Objective:
- To determine the contents of unknown solution, qualitatively and
quantitatively.
Abstract:

Chromatography is an extremely powerful tool used in analysis. It has different ways

to be done. In this experiment we used GLC to explore three different components at
different concentrations, and to study their retention time and their peak shape. For
instance, the retention time of chloroform, average of three trials, was 1.39 min. After
that, we plotted the peak area versus concentration, for the three components. Finally
for the unknown, we injected a 3 micro-liter of unknown solution, and according to
the retention time and the peak areas obtained, the components are identified and
their concentration is estimated.

Theory:
All forms of chromatography involve a stationary phase and a mobile phase. In all the
other forms of chromatography you will meet at this level, the mobile phase is a
liquid. 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.

How fast a particular compound travels through the machine will depend on how
much of its time is spent moving with the gas as opposed to being attached to the
liquid in some way.

 Injection of the sample

Very small quantities of the sample that you are trying to analyze are injected into the
machine using a small syringe. The syringe needle passes through a thick rubber disc
(known as a septum) which reveals itself again when the syringe is pulled out.

The injector is contained in an oven whose temperature can be controlled. It is hot

enough so that the entire sample boils and is carried into the column as a gas by the
helium (or other carrier gas).

 How the column works:

The packing material

There are two main types of column in gas-liquid chromatography. One of these is a
long thin tube packed with the stationary phase; the other is even thinner and has the
stationary phase bonded to its inner surface.

To keep things simple, we are just going to look at the packed column.

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The column is typically made of stainless steel and is between 1 and 4 metres long
with an internal diameter of up to 4 mm. It is coiled up so that it will fit into a
thermostatically controlled oven.

The column is packed with finely ground diatomaceous earth, which is a very porous
rock. This is coated with a high boiling liquid - typically a waxy polymer.

 The column temperature

The temperature of the column can be varied from about 50°C to 250°C. It is cooler
than the injector oven, so that some components of the mixture may condense at the
beginning of the column.

In some cases, as you will see below, the column starts off at a low temperature and
then is made steadily hotter under computer control as the analysis proceeds.

How separation works on the column

One of three things might happen to a particular molecule in the mixture injected into
the column:

 It may condense on the stationary phase.

 It may dissolve in the liquid on the surface of the stationary phase.
 It may remain in the gas phase.

None of these things is necessarily permanent.

A compound with a boiling point higher than the temperature of the column will
obviously tend to condense at the start of the column. However, some of it will
evaporate again in the same way that water evaporates on a warm day - even though
the temperature is well below 100°C. The chances are that it will then condense again
a little further along the column.

Similarly, some molecules may dissolve in the liquid stationary phase some
compounds will be more soluble in the liquid than others. The more soluble ones
will spend more of their time absorbed into the stationary phase; the less soluble ones
will spend more of their time in the gas.

The process where a substance divides itself between two immiscible solvents
because it is more soluble in one than the other is known as partition. Now, you
might reasonably argue that a gas such as helium can't really be described as a
"solvent". But the term partition is still used in gas-liquid chromatography.

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You can say that a substance partitions itself between the liquid stationary phase and
the gas. Any molecule in the substance spends some of its time dissolved in the liquid
and some of its time carried along with the gas.

 Retention time

The time taken for a particular compound to travel through the column to the detector
is known as its retention time. This time is measured from the time at which the
sample is injected to the point at which the display shows a maximum peak height for
that compound.

Different compounds have different retention times. For a particular compound, the
retention time will vary depending on:

 the boiling point of the compound. A compound which boils at a temperature

higher than the column temperature is going to spend nearly all of its time
condensed as a liquid at the beginning of the column. So high boiling point
means a long retention time.
 the solubility in the liquid phase. The more soluble a compound is in the
liquid phase, the less time it will spend being carried along by the gas. High
solubility in the liquid phase means a high retention time.
 the temperature of the column. A higher temperature will tend to excite
molecules into the gas phase - either because they evaporate more readily, or
because they are so energetic that the attractions of the liquid no longer hold
them. A high column temperature shortens retention times for everything in
the column.

For a given sample and column, there isn't much you can do about the boiling points
of the compounds or their solubility in the liquid phase - but you do have control
over the temperature.

The lower the temperature of the column, the better the separation you will get - but
it could take a very long time to get the compounds through which are condensing at
the beginning of the column!

On the other hand, using a high temperature, everything will pass through the
column much more quickly - but less well separated out. If everything passed
through in a very short time, there isn't going to be much space between their peaks
on the chromatogram.

The answer is to start with the column relatively cool, and then gradually and very
regularly increase the temperature.

At the beginning, compounds which spend most of their time in the gas phase will
pass quickly through the column and be detected. Increasing the temperature a bit
will encourage the slightly "stickier" compounds through. Increasing the temperature
still more will force the very "sticky" molecules off the stationary phase and through
the column.

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  The detector

There are several different types of detector in use. The flame ionization detector
described below is commonly used and is easier to describe and explain than the
alternatives.

 A flame Ionization Detector

In terms of reaction mechanisms, the burning of an organic compound is very

complicated. During the process, small amounts of ions and electrons are produced in
the flame. The presence of these can be detected.

The whole detector is enclosed in its own oven which is hotter than the column
temperature. That stops anything condensing in the detector.

If there is nothing organic coming through from the column, you just have a flame of
hydrogen burning in air. Now suppose that one of the compounds in the mixture you
are analyzing starts to come through.

As it burns, it will produce small amounts of ions and electrons in the flame. The
positive ions will be attracted to the cylindrical cathode. Negative ions and electrons
will be attracted towards the jet itself which is the anode.

This is much the same as what happens during normal electrolysis.

At the cathode, the positive ions will pick up electrons from the cathode and be
neutralized. At the anode, any electrons in the flame will transfer to the positive
electrode; and negative ions will give their electrons to the electrode and be
neutralized.

This loss of electrons from one electrode and gain at the other will result in a flow of
electrons in the external circuit from the anode to the cathode. In other words, you get
an electric current.

The current won't be very big, but it can be amplified. The more of the organic
compound there is in the flame, the more ions will be produced, and so the higher the
current will be. As a reasonable approximation, especially if you are talking about
similar compounds, the current you measure is proportional to the amount of
compound in the flame.

 Disadvantages of the flame Ionization Detector

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The main disadvantage is that it destroys everything coming out of the column as it
detects it. If you wanted to send the product to a mass spectrometer, for example, for
further analysis, you couldn't use a flame ionization detector.

 Interpreting the output from the detector

The output will be recorded as a series of peaks - each one representing a compound
in the mixture passing through the detector. As long as you were careful to control the
conditions on the column, you could use the retention times to help to identify the
compounds present - provided, of course, that you (or somebody else) had already
measured them for pure samples of the various compounds under those identical
conditions.

But you can also use the peaks as a way of measuring the relative quantities of the
compounds present. This is only accurate if you are analyzing mixtures of similar
compounds - for example, of similar hydrocarbons.

The areas under the peaks are proportional to the amount of each compound which
has passed the detector, and these areas can be calculated automatically by the
computer linked to the display. The areas it would measure are shown in green in the
(very simplified) diagram.

Note that it isn't the peak height that matters, but the total area under the peak. In this
particular example, the left-hand peak is both tallest and has the greatest area. That
isn't necessarily always so.

There might be a lot of one compound present, but it might emerge from the column
in relatively small amounts over quite a long time. Measuring the area rather than the
peak height allows for this.

Requirements:
- Volumetric Flasks.
- Pipettes.
- Measuring Cylinders.
- Funnels.

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Data Collected:
Table.1.Measured Retention time for three components.
Retention Time (min)
Concentration Acetone Chloroform Toluene
5% 0.72 -
10 % 0.76 1.4
15 % 0.8 -
20 % - 1.4
30 % - 1.37
Average 0.76 1.39

Table.2.Measured heights and widths from GLC data for

Chloroform. Chloroform ( in Acetone)
Concentration Height(cm) Width(cm) Area (cm2)
10 2.6 0.7 0.91
20 4.2 1.8 3.78
30 5.1 2.5 6.375

Table.3.Measured heights and widths from GLC data for Toluene.

Toluene ( in Acetone )
Concentration Height (cm) Width(cm) Area (cm2)
5 4.3225 1.3 6.65
10 9.45 1.8 10.5
15 16.7325 2.3 14.55

Table.4.Measured heights and widths form GLC data for Acetone.

Acetone ( in Toluene )
Concentration Height (cm) Width (cm) Area (cm2)
5 0.49 0.4 2.45
10 2.2 0.4 11
15 2.98 0.4 14.9

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Results:

Chloroform
7
6 f(x) = 0.27 x − 1.78
R² = 1
5
Peak Area (cm2)

Peak Area(cm2) Vs.

4 Concentration
3 Linear (Peak Area(cm2) Vs.
Concentration)
2
1
0
5 10 15 20 25 30 35
Concentration

Figure.1. Plot ( Peak Area Vs. Concentration ) for Chloroform.

Toluene
18
16 f(x) = 0.81 x + 0.68
14 R² = 0.99
Peak Area (cm2)

12 Peak Area (cm2) Vs.

10 Concentration
8 Linear (Peak Area (cm2)
6 Vs. Concentration)
4
2
0
4 6 8 10 12 14 16 18 20 22
Concentration

Figure.2. Plot ( Peak Area Vs. Concentration ) for Toluene.

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 Acetone ( in Toluene)
3.5
3 f(x) = 0.25 x − 0.6
Peak Area (cm2)
2.5 R² = 0.96
Peak Area (cm2) Vs.
2 Concentration
1.5 Linear (Peak Area (cm2) Vs.
Concentration)
1
0.5
0
4 6 8 10 12 14 16
Concentration

Figure.3. Plot ( Peak Area Vs. Concentration ) for Acetone.

For the Unknown:

We have three peaks, each one with different retention time correspond to one of the three
compounds we have; Acetone, Toluene and Chloroform.

And according to the retention time , the first peak is Acetone , the second is Chloroform and
the third is Toluene.

Area "Triangle" = (0.5) x Width x Height.

Area (cm2)
Acetone Chloroform Toluene
Height (cm) 16.1 Height (cm) 4.2 Height (cm) 3.25
Width (cm) 0.6 Width (cm) 0.7 Width (cm) 0.8
Area 4.83 Area 1.47 Area 1.3
Total Area = 7.6 cm2
% Area 63.55 %Area 19.34 % Area 17.11

Concentration of Acetone in the Unknown:

Area = 4.83 cm2 = y

From (fig.3)

y = 0.249x - 0.6 ----solving for x

x = 21.8 % (concentration).

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Similarly for Toluene and Chloroform;

Toluene ( y =1.3 cm2 )

From (Fig.2)

y = 0.813x + 0.681------solving for x

x=0.76 % (concentration).

Chloroform ( y =1.47 cm2 )

From (Fig.1)

y = 0.273x - 1.776------solving for x

x=11.89 % (concentration).

Then, in 3 (micro-liter) we have: 21.8 % Acetone, 0.76 % Toluene and 11.89 % Chloroform.

Conclusion and Recommendation:

Finally ,in this experiment we studied how the Chromatography is used and how
does it work .And we concluded on one hand ,that Chromatography is almost perfect
for identifying an unknown solution components, on the other hand ,the
concentration is also can be can obtained from the peak areas. Furthermore, this is
why Chromatography is used in different fields, even in forensics.

References:
1- http://www.chemguide.co.uk/analysis/chromatography/gas.html.
2- SKOOG/WEST/HOLLER, Fundamentals Of Analytical Chemistry,
Saunders HBJ , 6th Edition, 1992.

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