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STUDENT ID:
2014647344
PARTNERS NAMES: 1.
2.
3.
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
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.
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.
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.
D.
Sample calculation:
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.