CHM 260 LABORATORY REPORT

Experiment Number: 5

Title: Gas Chromatography (GC)

Name: Nor Syafikah binti Mohd Nasir

Student no.: 2015440114

Date of Report Submission: 15th December 2017

Lab Partner’s Name: Aqilah binti Anuar

Lyana Fasihah binti Ahmad Sobri

Nur Atiqah Farzana binti Zaini

Lecturer’s Name: Miss Hanani binti Yazid

EXPERIMENT 5: Gas Chromatography (GC)

OBJECTIVES

1. To determine the retention times tR of n-butanol and 2-propanol.

2. To identify the components present in a standard mixture based on the t R.
3. To identify the components present in an unknown sample.
4. To determine the effect of temperature on tR and Rs.

INTRODUCTION

Gas chromatography is an analytical separation technique that analyses compounds

eluted from the column, which carried through the column by a gas mobile phase.
The carrier gas in our experiment was nitrogen. We used nitrogen because it is
inexpensive but gives reduces sensitivity. A detector was located at the end of the
column in gas chromatography to measure the solutes that elute from it. In order to
obtain an ideal chromatogram, there are many parameters that need to be adjusted.
Ideal parameters give chromatograms with the high resolution in the short amounts
of time. There are many parameters that can affect both the elution time and the
resolution but for this experiment, we focused on effects of temperature and
pressure. In gas chromatography, an increase in temperature is only favoured up to
a certain point. The increase temperature reduces the time of elution for all the
analytes, but after a certain point, it causes a decrease in the resolution. Pressure
also needs to be controlled in order to obtain a favourable chromatogram. High
pressure causes backflow which causes elution time to increase. Since the
temperature and pressure are dependent on each other and it would take too long to
figure out mathematically, the parameters were optimized in our experiment with the
Simplex Optimization software. The Simplex Optimization software can only be used
if the parameters being optimized are dependent on each other. This software used
data from test runs to calculate what it thought to be a good set of parameters. The
results from those parameters were then reentered into the system to make a better
measurement. In our experiment, we performed as many trials as possible in the
time allotted. In the end, we used the parameters that gave a chromatogram that
looked to have the highest resolution in the shortest amount of time.
APPARATUS

Syringe

Beaker

Dropper

CHEMICALS

2-propanol

n-butanol

Standard mixture of 2-propanol and n-butanol (1:1 ratio)

Unknown sample

PROCEDURE

A. Sample Handling
1. The syringe was rinsed before it was filled with the sample. The volume of
the sample may be more than the required volume. It was make sure that
no bubbles in the syringe. Tap the syringe gently to remove any air
bubbles.
2. The syringe was hold vertically, needle up and the plunger was pushed to
the required volume at eye level. The excess sample was removed using a
tissue.
B. Operation of the GC
1. The sample was injected into the septa.
2. The oven column was checked. The instrument was make sure is in the off
mode.
3. The instrument was switched on. The warning tone will ring if the gas is
not enough.
4. “Instrument 1 online” was clicked in window screen. The instrument is
allowed to setup itself.
5. After setup is completed, “method” was click → “edit entire method” →
“ok”.
6. The title was entered and click ok.
7. To select injection, click “manual”. To select injection location, click “back”.
At instrument, edit inlet, choose “back” and keep in information.
8. If the sample is in high concentration, “split” mode was chosen. If sample
is in low concentration or in small quantities, “splitless” was chosen in inlet.
9. Go to column section, “column no. 2” was chosen. At inlet column, “back”
was chosen. At mode column, “constant pressure’ was chosen and at
outlet column, “ambient” was chosen.
10. Go to detector column. FID was used.
11. The parameter was injected. (time:5min, 1µL).
12. After doing all of the above, “ok” was clicked.
13. “apply” was clicked → ok → signal detail → ok → edit integration event→
ok → specify report → ok → run time checklist → ok.
14. Go to “file”, click “save”, “method”. Keep in the command.
15. During the injection, make sure that no bubbles appear in the syringe.
16. Go to “run control”, click “sample info”. The name was set in the “operator
name” column.
17. Set signal 2, the sample name was filled in.
18. Click “run method” and the instrument will wait for injection.
19. The sample was injected quickly and the button “start” was pressed
quickly on the instrument. Then, the syringe was quickly pulled from the
septa.
20. Go to data analysis, click “file” then “load signal”.
21. “calibration” was entered, click “new calibration table”. The name was
entered and the document was printed.
22. For the next sample, “data analysis” was clicked and steps 17-21 were
repeated.
QUESTIONS

1. State the type of compounds which are suitable for analysis using GC.
The type of compounds which are suitable for analysis using GC are
hydrocarbons, halogenated compounds, compounds containing
halogens, sulphur or nitrogen and organic compounds.

2. Why is FID a suitable detector for this analysis?

FID is a suitable detector for this analysis because FID is used to detect
hydrocarbons. It will not detect non-carbon containing compounds. This
detector responds to all organic compounds except formic acid and the
response is highest for hydrocarbons. Besides that, the detector is
insensitive to H2, O2, N2, SiCl4, SO2, NO2, H2O and NH3. FID requires
hydrogen supply and oxygen supply. Lastly, FID exhibits high
sensitivity.

3. List two factors which can increase the efficiency of a GC column.

Factors which can increase the efficiency of a GC column is lengthen
the column. Besides that, reducing the packing size of column can
increase the efficiency of a GC column.

DISCUSSION

In the experiment, retention time plays big role in analysing the sample in gas
chromatography. For the first part of the experiment, retention times are
differentiated according to the number of carbon, molecular weight and the boiling
point of the samples. Graphs plotted from these readings that obtained from the
chromatography shown that log tr is proportionally increase when number of carbon
is increased. It goes the same with molecular weight and boiling point. Log t r
increase when the molecular weight and boiling point of the compound are
increased. This data concluded to the relationship that when the number of carbon in
samples tested increased, their molecular weight also increased and so with their
boiling point. Therefore, the retention time, time for a maximum of symmetrical peak
to occur is also increased. Next, the temperature changed during the experiment of
GC effected on tr and Rs. The higher the temperature was set, the longer it take for
the compounds to elute. Therefore, the resolution will increase resulting the
increasing of the efficiency. Based on the data obtained, when the temperature was
70°C, the retention time for the compounds standard mixture is 1.557 min for 2-
propanol and 1.999 for n-butanol. For the second injection, the retention time of 2-
propanol was 1.561 min and n-butanol was1.995min. For the unknown sample, its
retention time was 1.539min, so it shows that unknown sample was 2-propanol.
Then, the temperature was increased to 100°C and the standard mixture was
injected. The graph plotted shows to peak with retention time 1.586 min and 1.725
min. Based on the retention time obtained, we can know that 1.585 min was
retention time for 2-propanol while 1.725 min was retention time for n-butanol. Next,
when the temperature was 140°C, the retention time for 2-propanol peak was 1.677
min and 1.732 min for n-butanol. During the experiment was conducted, there were
several precautions to be taken in order to minimize errors in the data. Firstly, shake
all the solution adequately to ensure dissolution for 5min before injection into
chromatograph. Then, make sure the column is not clogged, it maybe cause by the
precipitation of proteins in the column caused by removal of stabilizing agents during
fractionation. Beside that, the syringe was rinse and clean before and after each
sample was injected to prevent any contaminant and other solution that will affect the
process. Lastly, avoid formation of bubbles during the experiment because it will
affect the reading in the column while the measurement was taken.

CONCLUSION
At the end of the experiment, we can determine the retention time of n-butanol and
2-propanol which are 1.999 min and 1.557 min respectively. Beside that, we
managed to identify the components present in a standard mixture based on the t r.-
Then, the components present in the unknown sample was identified. The
component present can be known based on the retention time obtained in the graph.
It shows that 2-propanol was present as its retention time was 1.539 min. Last but
not least, we had learned the effect of temperature on t r and Rs. As the temperature
increase, the retention time, tr will increase. So the Rs will increase as well.
REFRENCES
1. Ariffin Z., 2016, Basic Instrumental Analysis Laboratory Experiments for An
Introductory Course in Instrumental Analysis, Gas Chromatography
Experiment 5.
2. https://www.scribd.com/doc/38677786/Experiment-of-Gas-Chromatography,
retrived on 9 December 2017, 5.08pm.