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(EXPERIMENT 1)

GAS CHROMATOGRAPHY (GC), OPTIMIZATION


OF FLOW RATE AND COLUMN TEMPERATURE
NAME:

MOHAMAD NOR AMIRUL AZHAR BIN KAMIS

STUDENT ID:

2014647344

PARTNERS NAMES: 1.

MOHAMAD HAMIZAN BIN MOHD ISA

2.

MOHAMAD SHAFIQ BIN PARMAN

3.

MOHAMAD AZMIZAM BIN MOHAMAD


NOOR

DATE OF EXPERIMENT:

16/10/2014

DATE OF SUBMISSION:

8/12/2014

INTRODUCTION:
Gas chromatography is a term used to describe the separation techniques used to
analyse volatile substances in the gas phase. 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 such as helium or hydrogen. The mobile phase is a
chemically inert gas that serves to carry the molecules of the analyte through the
heated column. The mobile phase goes through a glass column packed with silica
that is coated with a liquid. Materials that are less soluble in the liquid will elute faster
than the material with greater solubility. The rate of which compound travels in the
column depends on the volatility of compound, column temperature, flow rate of the
mobile phase and the length of column. Low boiling point compound (volatile) will
elutes faster than the high boiling point compound. High column temperature and
high mobile phase flow rate will reduce the time of solute in the column. Longer
column length will take longer time to elute all compounds but will give better
separation due to high number of theoretical plates and smaller plate height. The
liquid stationary phase is adsorbed onto a solid inert packing or immobilized on the
capillary tubing walls in a thin layer. In a capillary column, the tubing walls are coated
or bonded with the stationary phase liquid. A typical gas chromatograph consists of
an injection port, a column, carrier gas flow control equipment, ovens and heaters for
maintaining temperatures of the injection port and the column, an integrator chart
recorder and a detector.
The technique propose in this experiment has been employed in isothermal gas
chromatography with split injection since, the concentration of the analyte is high.
Isothermal gas chromatography is an analysis where the column temperature is
remained constant throughout the analysis. Resolution (Rs) is used to describe how
well the species in the mixture are separated. Ideal resolution is 1.5 that indicates
adequate separation between species. Lower than 1.5 resolutions is when the two
species is not completely separated while too high resolution will need longer
analysis time. The objective of this experiment is to explore gas chromatography;
including the concepts of retention time and resolution using a mixture of methyl
esters; methyl laurate, methyl myristate, methyl palmitate, methyl stearate and
methyl linoleate. The effects of column temperature and flow rate on the separation
of these compounds will be investigated.

Where

t : retention time of analytes


W: peak base width

EXPERIMENTAL:
a. Isothermal elution was set up with condition of:
Injection port: split (40:1)
Injection port temperature: 250C
Column temperature: varies (170C, 190C, 210C)
Carrier gas flow rate: varies (30m/s, 50m/s, 70m/s)
Detector temperature: 250C
b. Effect of carrier gas flow rate on isothermal GC separation of methyl
ester:
1. The standard mixture was injected with 0.4L at 210C isothermally and
30m/s of gas flow rate.
2. For the second injection with 0.4L volume of standard mixture, the gas
flow rate was increased to 50m/s with same temperature of 210C.
3. The same procedure was repeated with 70m/s gas flow rate.
4. The resolution of 3 different flow rates at the same temperature was
calculated and compared to determine which gas flow rate give the better
separation of the compounds in the standard mixture.
c. Effect of column temperature on the isothermal GC separation of methyl
ester:
1. By using the best gas flow rate from the above procedure, the same
standard mixture with 0.4L was injected into the column isothermally at
170C and followed by 190C.
2. The resolution of each chromatogram was calculated and was compared
based on the resolution and the analysis time.
d. Identification of components in methyl esters mixture:
1. The standard individual methyl ester compounds were injected with 0.4L
volume using the optimized GC conditions.
2. The standard individual methyl ester compounds are methyl laurate,
methyl myristate, methyl palmitate, methyl stearate and methyl linoleate.

RESULT AND DISCUSSION:


*Calculation of resolution is based on peak 2 and 3 as references.
A.

Effects of the variation of the gas flow rate on the resolution:

Condition

Injection

1
30m/s,
210C

2
1

50m/s,
210C

2
1

70m/s,
210C

Retention
time of
peak 2 and
3 (min)
2.812,
4.015
2.807,
4.003
1.684,
2.412
1.680,
2.406
1.190,
1.706
1.199,
1.714

Peak width
of peak 2
and 3 (min)

Resolution

0.1070,
0.2052
0.1084,
0.2056
0.0755,
0.1480
0.0746,
0.1447
0.0578,
0.1137
0.0509,
0.1042

7.71

Average
resolution

7.67
7.62
6.51
6.57
6.62
6.02
6.33
6.64

The optimized separation time of methyl esters mixture is at 70m/s of gas flow rate.
B.

Effects of the variation of column temperature at optimized column


temperature on the resolution:

Condition

Injection

Retention
Peak width Resolution
Average
time of
of peak 2
resolution
peak 2 and and 3 (min)
3 (min)
1
2.311,
0.1588,
8.99
70m/s,
4.655
0.3626
9.05
170C
2
2.302,
0.1513,
9.10
4.656
0.3661
1
0.505,
0.0868,
7.65
7.80
70m/s,
2.519
0.1783
190C
2
10548,
0.0842,
7.95
2.596
0.1793
1
1.190,
0.0578,
6.02
70m/s,
1.706
0.1137
6.33
210C
2
1.199,
0.0509,
6.64
1.714
0.1042
The optimized column temperature at 70m/s of gas flow rate is 210C column
temperature because it produced the resolution nearest to the ideal resolution value
that is 1.5 and also shorter analysis time.

C.

Retention time of standard compounds of the methyl esters:


Standard compound
Methyl laurate
Methyl myristate
Methyl palmitate
Methyl linoleate
Methyl stearate

D.

Retention time (min)


1.199
1.721
2.815
4.667
5.070

Sample calculation:

*condition of 70m/s gas flow rate at 210C:


Rs (2,3) =

= 6.02 (first trial)


Rs (2,3) =
= 6.64 (second trial)
Average resolution (2,3) =
= 6.33
The variation of the mobile phase flow rate will affect the retention time of the
compounds in which slow mobile phase flow rate will give better separation but very
long analysis time. In contrast, high flow rate will shorten the analysis time but will
cause broadening due to the mass transfer (C-term) in Van Deemter plot, because
the solute does not completely interact with the stationary phase. To reduce the
analysis time and to produce better separation, the optimum gas flow rate must be
used. In this experiment, the optimum mobile phase flow rate is 70m/s that give good
resolution of 6.33 compared to the others that are far from the ideal resolution value
that is 1.5. The column temperature also affects the separation resolution and the
analysis time. High column temperature will give short analysis time but some of the
earlier peaks may be overlapped while low column temperature produces better
separation but will take very long analysis time. The optimum column temperature
must be used in order to separate each compounds adequately. In this case, 210C
of column temperature is the best temperature to separate each of the compounds.
Based on this experiment, the best condition to separate the methyl ester mixture is
by using 70m/s gas flow rate at 210C column temperature that will give adequate
separation between compounds and also shorter analysis time.

Optimum flow rate and optimum column temperature will produce better separation,
high efficiency, good resolution and short analysis time for the separation. Since the
separation of gas chromatography is based on the boiling point of the compounds, it
can be concluded that methyl laurate has lowest boiling point followed by methyl
myristate and highest boiling point is the methyl palmitate.

CONCLUSION:
The optimized condition for the separation of the methyl esters is 70m/s of gas flow
rate and 210C of column temperature. The first peak after the solvent peak is
corresponds to methyl laurate followed by methyl myristate and methyl palmitate.

REFERENCES:
1. Norashikin S., Ruziyati T., Mardiana S. (2012), Analytical Separation
Methods Laboratory Guide (2nd edition), 3/10/2014.
2. Y. Guillaume and C. Guinchard (January 1997), France, Prediction of
Retention Times, Column Efficiency, and Resolution in Isothermal and
Temperature-Programmed Gas Chromatography: Application for Separation
of
Four
Psoralens,
3/10/2014,
http://chromsci.oxfordjournals.org/content/35/1/14.full.pdf.
3. Gas
Chromatography,
4/10/2014,
http://chemwiki.ucdavis.edu/Analytical_Chemistry/Instrumental_Analysis/Chro
matography/Gas_Chromatography.
4. Mardiana Saaid, Gas Chromatography Lecture Notes, 5/10/2014.