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CHE 314 ANALYTICAL CHEMISTRY LABORATORY

GROUP: WEDNESDAY

EXPERIMENT 5

TITLE:

WEEK : 4

DATE: 25 03 2015

NAME: KOOGOTSITSE SEKHOBE

STUDENT NUMBER: 200604183

ANALYSIS OF CHLOROFORM
CHROMATOGRAPHY

AND

CARBON

TETRACHLORIDE

BY

GAS

INTRODUCTION
Gas Chromatography (GC) is used to separate volatile components of a mixture. In gas
chromatography, the components of a sample are dissolved in a solvent and vaporized in order to
separate the analytes by distributing the sample between two phases: a stationary phase and a
mobile phase. The mobile phase is a chemically inert gas such as helium or nitrogen that serves
to carry the molecules of the analyte through the heated column. It is within the column that
separation of the components takes place. Molecules partition between the carrier gas (the
mobile phase) and the high boiling liquid (the stationary phase) within the GC column. Therefore
gas chromatography is one of the sole forms of chromatography that does not utilize the mobile
phase for interacting with the analyte. The stationary phase consists of a packed column where
the packing or solid support itself acts as stationary phase, or is coated with the liquid stationary
phase (high boiling polymer). Most analytical gas chromatographs use capillary columns, where
the stationary phase coats the walls of a small-diameter tube directly i.e., 0.25 m film in a 0.32
mm tube (Mustafa, 2012). After components of the mixture move through the GC column, they
reach a detector. Ideally, components of the mixture will reach the detector at varying times due
to differences in the partitioning between mobile and stationary phases. The detector sends a
signal to the chart recorder which results in a peak on the chart paper. The area of the peak is
proportional to the number of molecules generating the signal.
In this experiment, Carbon tetrachloride (CCl4) and Chloroform (CHCl 3) mixture were analyzed
using gas chromatography, the chromatograms obtained from the chromatograph were used to
plot a curve and the results were used to determine the amount in percentage of chloroform in the
unknown.
EXPERIMENTAL SECTION
Apparatus and Reagents
APPARATUS

CHEMICALS

Gas chromatograph with recorder


Volumetric flask (20 mL)
Syringe (L)
vile

Chloroform
Carbon tetrachloride
Hexane
acetone

Procedure
The volumetric flasks were first cleaned with hexane and then acetone before being used. The
standards were then prepared using the 20mL volumetric flasks as follows:

First volumetric flask: 14 L of chlororoform (CHC 3) was added and it was diluted to the mark
with acetone
Second volumetric flask: 2000 L of carbon tetetrachloride (CCl4) was added and the solution
diluted to the mark with acetone.
The solutions in the volumetric flasks were then transferred into the vile to make a standard
series of mixture of CHCl3 and CCl4 that contain 30, 40, 50, 60 and 70 % CHCl 3 by volume.
The total volume of each mixture was 1.5 mL (1500 L) in a vile therefore the volumes of how
much each solution contributed in each standard were as shown in the table below.
CHCl3 in %

volume of CHCl3 in L

volume of CCl4 in L

30

450

1050

40

600

900

50

750

750

60

900

600

70

1050

450

The flow rate of the gas chromatograph was adjusted to the optimum value for CHCl 3, the
temperature of the oven was programmed considering boiling points of the two compounds and
then the five mixtures were each run in the chromatograph and a chromatogram of each was
obtained. Only about 20 L of each sample was run in the chromatograph. The uncombined
solution of CHCl3 and CCl4 were also run. Split injection was used to inject the analytes into the
chromatograph injection port. The parameters set on the gas chromatograph instrument can be
summarized as follows:
Detector: Electron Capture detector

Carrier gas: Nitrogen

Column length: 30 m

column diameter: 320 m film

Mode of injection: split injection

Split ratio: 100:1

Injection temperature: 150 oC

temperature programmed: 70 100 oC

Holding time: 2 minutes

Run time: 15 minutes

Flow rate: 0.5 ml/minute


RESULTS ANALYSIS

GRAPH OF AREA RATIO UNDER THE PEAK OF CHLOROFORM TO TETRACHLORIDE AGAINST 1/R

1.8
f(x) = 0.77x - 0.86
R = 0.89

1.6

1.4

1.2

1
area ratio (CHCl/CCl)
0.8

0.6

0.4

0.2

0
1

1.5

2.5

3.5

1/R (R = volume fraction of tetrachloride in the mixture)

Figure: Graph showing a straight line (the line of best fit) as plotted from area ratio of
CHCl3/CCl4 against 1/R
CALCULATIONS OF % CHCl3 IN THE UNKNOWN

Peak area of CHCl3 in the unknown = 2.135 106 Hz*s


Peak area of CCl4 in the unknown = 1.746 106 Hz*s
Peak area ratio of CHCl3/CCl4 in the unknown = 2.135 106 Hz*s/ 1.746 106 Hz*s
= 1.223
Hence: Using the obtained value above to find 1/R from the graph

1
R

= 2.7

Where R is the volume fraction of tetrachloride (CCl4) in the mixture


R=

1
2.7

= 0.37 or

37
100

Therefore:
Volume fraction of chloroform (CHCl3) = 1- 0.37
= 0.63 or

63
100

= 63%

Hence the % CHCl3 in the unknown was 63%

QUESTIONS
1. Explain the elution order.
The separation of compounds was based on the different strengths of interaction of the
compounds with the stationary phase. The stronger the interaction is, the longer the compound
interacts with the stationary phase, and the longer the retention time. Therefore CHCl3 (polarity
index = 3.1) will elute first because it is more polar than CCl4 (polarity index = 1.6) while the
stationary phase is non polar which means that CCl4 which is less polar and will interact more
with the stationary phase hence eluting last. Polarity index is a relative measure of the degree of
interaction of the solvent with various polar solutes (Byers, 2003).
2. Do you expect CH2Cl2 to elute faster or slower than CHCl3?
CH2Cl2 (dichloromethane) will elute slower than CHCl3 because CH2Cl2 is less polar than CHCl3
hence it will interact more with the stationary phase than CHCl3. That is more intermolecular
interactions with stationery phase are expected of CH2Cl2 than chloroform. Polarity index of
CH2Cl2 is 3.1 while that of CHCl3 is 4.1 (Byers, 2003).

DISCUSSION
It is important to note that the separation of compounds using gas chromatography was based on
the different strengths of interaction of the compounds with the stationary phase. If the polarity
of the stationary phase and compound are similar, the retention time increases because the
compound interacts stronger with the stationary phase. As a result, polar compounds have long
retention times on polar stationary phases and shorter retention times on non-polar columns
using the same temperature. An excessively high column temperature results in very short
retention time but also in a very poor separation because all components mainly stay in the gas
phase. However, in order for the separation to occur the components need to be able to interact
with the stationary phase. If the compound does not interact with the stationary phase, the
retention time will decrease. At the same time, the quality of the separation deteriorates, because
the differences in retention times are not as pronounced anymore. The best separations are
usually observed for temperature gradients, because the differences in polarity and in boiling
points are used here (Basset, Denney, Jeffery, & Mendham, 1989).
Carrier gas flow rate - A high flow rate reduces retention times, but a poor separation would be
observed as well because the components have very little time to interact with the stationary
phase and are just being pushed through the column. Also to note is amount of material injected,
the peaks in the chromatogram display a Gaussian curve therefore if too much of the sample is
injected, the peaks show a significant tailing, which causes a poorer separation. Hence most GC
instruments are operated in split-mode to prevent overloading of the column and the detector,
this is why split mode was used in this experiment. The split-less mode is only used if the sample
is extremely low in concentration in terms of the analyte.
Separation of a 30% CHCl3 and 70% CCl4 mixture was first tried using isothermal programming
of the oven temperature, then it was done using temperature programming in order to compare
the two methods. In isothermal programming, the temperature of the column is held constant
throughout the entire separation. The optimum column temperature for isothermal operation is
about the middle point of the boiling range of the sample. However, isothermal programming
works best only if the boiling point range of the sample is narrow. If a low isothermal column
temperature is used with a wide boiling point range, the low boiling fractions are well resolved
but the high boiling fractions are slow to elute with extensive band broadening. If the
temperature is increased closer to the boiling points of the higher boiling components, the higher
boiling components elute as sharp peaks but the lower boiling components elute so quickly there
is no separation. The chromatograms obtained hence showed poorly resolved peaks. For these
reasons, isothermal temperature conditions are only suitable for a limited number of analyses
In temperature programming method, the column temperature is either increased continuously or
in steps as the separation progresses, that is a temperature program involves heating the oven at a
controlled rate during the run. The method is well suited to separating a mixture with a broad

boiling point range. The analysis begins at a low temperature to resolve the low boiling
components and increases during the separation to resolve the less volatile, high boiling
components of the sample. This allows the faster analysis of solutes with dissimilar retention,
and there is very little peak broadening with an increase in retention. The only disadvantage of a
temperature program is the cool down time between analyses and also there are no ways of
finding the best temperature program for an analysis. Usually to achieve satisfactory peak
resolution, efficiency may be improved, optimizing the carrier gas average linear velocity,
improving injector efficiency, or using a more efficient column dimension may provide the
desired resolution. Therefore for this mentioned reasons temperature programming was chosen
as the right method for operating the oven in all the standards.
The results obtained showed that 63% of chloroform is contained in the unknown whereas 37%
of the unknown mixture was tetrachloride. However the calculations were not including the other
substances (contaminants) that are shown to have been there by the chromatograms obtained.
This meaning that there was some improper handling of the standards during preparation which
led to contaminations of the sample. The analytes might have been also contaminated during
injection into the gas chromatograph by the syringe which may have had strongly sorbing
substances on the inner surface. The adsorbed material could be displaced by the next selected
sample with a differing chemical composition which will falsify this next sample (Riley &
Szecsody, 2005).
Nonetheless, the errors did not have much effect on the results obtained as the sample results in
the chromatograms could be easily identified. The peak areas of the two samples were obtained
and used to plot a curve of Area ratio CHCl3/CCl4 against 1/R which was used to find the results
stated above. The curve plotted was precise since R2 obtained in the graph was 0.89 which is
closer to 1(one).
An analysis of samples using gas chromatograph is advantageous in that it is fast, requires only
very small samples with little preparation and the equipment is not very complex but reliable.
Gas chromatograph is also highly sensitive but non-destructive and therefore it is good at
separating complex mixtures into components, it also has a very high precision and has the
sensitivity to detect volatile organic mixtures of low concentrations. The disadvantages of the gas
chromatography are that it is limited to volatile samples, not suitable for thermally labile
compounds and requires spectroscopy (usually mass spectrometer) to confirm the peak identity.
CONCLUSION
Gas chromatography was used to analyze chloroform and carbon tetrachloride and a calibration
curve from the analysis was constructed hence used to calculate the percentage of chloroform in
the unknown. The percentage of chloroform in the unknown therefore was found to be 63 %.

References
Basset, G., Denney, R., Jeffery, G., & Mendham, J. (1989). Vogel's textbook of
chemical analysis, fifth edition. New York: John Wiley and Sons.
Byers, A. J. (2003, 3 24). Phenomenex catalog. Retrieved 03 24, 2015, from
www.Phenomenex.com: http://www.chemical-ecology.net/java/solvents.htm
Mustafa, A. M. (2012). Advanced Gas Chromatography: Prgress in Agricultural,
Biochemical and Industrial Applications. Croatia: In Tech.
Riley, R., & Szecsody, J. (2005). Carbon Tetrachloride and Chloroform Partition
Coefficients Derived from Aqueous Desorption of Contaminated Hanford
Sediments. U.S.A: U.S Department of Energy.