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Discussion.

Gas chromatography (GC) is a column chromatography technique which the mobile phase is gas
and stationary phase is either an immobilized liquid or a solid. This experiment is Perkin Elmer
Gas Chromatography which it is using gas as mobile phase and solid packed column as
stationary phase. GC is used to separate thermally stable volatile components of a mixture. In
this experiment, it is used to quantify the total Fatty Acid Methyl Ester (FAME) content and
Methyl Linolenate in each sample. This method was determined through the area peaks of each
component detected in chromatogram.
Fatty acids, esterified to glycerol, are the main constituents of oils and fats. The industrial
exploitation of oils and fats, both for food and oleochemical products, is based on chemical
modification of both the carboxyl and unsaturated groups present in fatty acids. Fatty acids are
almost entirely straight chain aliphatic carboxylic acids. The broadest definition includes all
chain lengths, but most natural fatty acids are C4 to C22, with C18 most common.
Gas chromatography was used to identify and measure the composition of fatty acids present in
the three samples of oil which were olive oil and palm oil. The composition of fatty acids present
was calculated with concentration of species from the reference standard of fatty acids methyl
ester (FAME). A FAME reference standard mixture is used in order to determine the retention
times and peak area of each component and it also used to identify components peaks in the
chromatogram through retention times obtained in standard.
Approximately 50 mg of sample was weighed and dissolved with 0.95 of hexane into 2 mL vial.
This mixture was shake thoroughly using vortex machine until a separation is seen. This method
is known esterification. This method purposely to form fatty acid methyl esters (FAMEs) from
fatty acids liberated so that the volatility is increased. Once the mixture is separated, the clear
solution is took out and dried by using addition of anhydrous sodium sulphate. This method is
known as extraction in which solute is extracted from one solvent by shaking it with a second
immiscible solvent. This method is to extract water from the sample and the extracted sample is
further cleaned up using anhydrous sulphate. 10 L of extract FAME is took out and diluted with
Hexane in 1 mL conical flask.

The samples were then analyzed by injecting 1 L of each sample into GC and recorded
chromatogram. The FAME contents were then calculated using integrated peak areas identified
in the samples and standard mixture. Fatty acid profile that is experimentally determined is
compared with literature source. Based on calculation, it showed that in 1 L of FAME sample
palm oil contains 1.227 g Palmitic, 0.169 g Stearic and 1.318 g Oleic.
The retention time of each methyl esters are shown in Table 3. In general, all components elute in
order of increasing carbon number and retention time. As shown in Table 3, Solvent front has
lowest retention time, 1.272 minutes and Oleic Methyl Ester has highest retention time, 4.401
minutes. Based on this, component with higher carbon number has higher affinity and less
mobile analytes as it has stronger interaction with stationary phase. Component with lower
carbon numbers has less affinity which resulted faster transported through the system in mobile
phase. Solvent front has lower carbon numbers, than Oleic Methyl Ester. Other than that, Oleic
Methyl Ester is unsaturated and have more than one double bonds which resulting in greater
polarity compared to Solvent front which is saturated.
The peak area of palm oil is 666663.52. Palmitic acid acid has many functions in cosmetics,
from detergent cleansing agent to emollient. In a cleanser, it has the potential to be drying
depending on what it is combined with and the cleansers pH level.
For the precaution step, while handling the procedure of gas chromatography. Few safety
measurements should be taken to ensure the safety in laboratories. Gas chromatography requires
handling compressed gases (nitrogen, hydrogen, argon, helium), and flammable and toxic
chemicals. Consult product MSDSs before using such hazardous products. Before injected the
solvent into Gas Chromatography, ensure not to let any bubbles in the syringes to prevent any
interference in the chromatograph that could resulting in extra peak present. Before supplying
hydrogen gas, confirm that all the connections are completed or the outlets are blocked off.
Failure to do so can cause serious accidents, such as fires or explosions.

Conclusion
The purpose of this experiment is to determine the composition of fatty acid
methyl ester by using the Gas Chromatography. From this experiment, it can be concluded that
each retention time represent each component. When the peak of the samples appear before the
standard peak or in other case the peak of certain component does not appear at all, it affects the
result. Thus concentration of each component cannot be calculated. This situation is due to
present of undesired organic component.
The experiment is about the determination fatty acid methyl ester (FAME) by oil
using the gas chromatography. The objectives of the experiment is achieved. The experiment
obtained three type of FAME which are methyl palmitate, methyl stearate and methyl linoleic.
The result using the retention time and peak area. The result shown olive oil having the better
result compared to the palm oil and used oil. The olive oil shows retention time at 5.072 and the
peak area at 2.39 for the methyl linoleic, the methyl stearate at 4.740 at time retention and the
peak area 47.02 and the methyl palmitate retention time at 6.072 and the peak area at 6.53.
Meanwhile the concentration is for the methyl palmitate at concentration 6.12x10-4, methyl
stearate at concentration 0.01 and methyl linoleic at concentration at 5.25x10 -4. The error can
reduced with accurately and accuracy.

CONCLUSION

As a conclusion, the composition of the fatty acid in the oil samples can be
determined by using gas chromatography analysis. Therefore for the determination of fatty
acid composition of oil samples it is very hard to obtain all the standard of fatty acids and
is difficult to identify some peaks. Based on the theory, each sample of oil must contain at
least three of the standard fatty acid. The product contain less saturated fatty acid is good for
human consumption. Yet, the consumer should avoid consume oil or fat from animal because it
contain the most saturated fatty acid. The higher amount of Trans Fatty Acids (TFA) and
saturated fatty acid demonstrated the poor quality of product in the market which is harmful to
the health of consumers.

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RECOMENDATION
In present study Gas Chromatography Mass Spectroscopy (GC-MS) can be used to

achieve more accurate peak identification. Great variation in the fatty acid composition has
been observed among different sample of fatty acids.

As conclusion, the objective to determine fatty acid methyl ester (FAME) in oil by
gas chromatography was achieved. The highest percentage in the sample is methyl
palmitate and followed by, methyl linoleic and methyl stearate. While for the concentration

of the sample the highest was methyl palmitate for the sample of Palm Oil and about the
same in Corn Oil and Olive Oil. The lowest concentration was Linolelaidic Acid Methyl
Ester in Corn Oil, but Trans-9 Elaidic Methyl Ester cannot be detected for Olive and Palm
Oil.
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RECOMMENDATION

The precaution that should be applied during run GC analysis is must make sure
there is no bubble inside the microliter syringe to avoid inaccuracy in measurement of the
samples before inject the sample or standard into the injection port. The microliter syringe
must be rinsed before and after inject the sample into the injection port for about three
times to ensure that no contamination occur during the analysis.