FACULTY OF APPLIED SCIENCES

LABORATORY REPORT

INSTRUMENTAL ANALYSIS OF FOOD

(FST606)

TITLE OF EXPERIMENT : DETERMINATION OF FATTY ACIDS IN COOKING OIL

USING GAS CHROMATOGRAPHY
NO. EXPERIMENT : 6
NAME OF STUDENT :
STUDENT ID :
GROUP : AS2465B
GROUP MEMBERS :
DATE OF EXPERIMENT : 23 OCTOBER 2023
DATE OF SUBMISSION : 30 OCTOBER 2023
LECTURER’S NAME :
INTRODUCTION
Fatty acids are referred to as carboxylic acids which are the building blocks of fats,
oils, and all other types of lipids aside from steroids. There are over 70 known fatty acids in
the world. They are typically unbranched, have an even number of carbon atoms (12–20 on
average), and may be divided into groups based on the presence and amount of carbon–
carbon double bonds (Libretexts, 2021). Several methods can be used to identify fatty acids
in oil and food. Fatty acid analysis has been carried out for more than a decade using
infrared spectroscopy, capillary electrophoresis, high-performance liquid chromatography
(HPLC), and gas chromatography (GC). In the meantime, GC has been employed to study
fatty acids for more than 50 years because it is a precise, sensitive, repeatable, and flexible
instrument to analyze complex fatty acids quickly and in an accurate manner. Although
several analytical methods have been applied to determine fatty acids, Gas Chromatography
– Flame Ionization Detector is still the most effective, and widely used method for identifying
and quantifying fatty acids in foods and essential oils (Alinafiah et al., 2021).

Gas chromatography is a common analytical method for separating and analyzing

gaseous and volatile substances. Gas Chromatography may be used for both qualitative and
quantitative analysis. To separate the analytes in gas chromatography, the sample is divided
into two phases: this can be regarded as a stationary phase and a mobile phase. The
sample is dissolved in a solvent and then evaporated to obtain its constituent parts. The
mobile phase (normally made up of a chemically inactive gaseous element such as nitrogen
or helium) carries the analyte molecules along the heated column (Kaur, 2020). The Gas
Chromatography method can also be referred to as gas–solid chromatography (GSC) or
gas–liquid chromatography (GLC) depending on the physical condition of the stationary
phase, which can be either solid or liquid. The primary methods of separation are partition
chromatography or adsorption. The separation of different components is achieved in GSC
by their adsorptive properties with respect to the solid stationary phase. If the analytes have
distinct distributions between the mobile and stationary phases, separation is achieved in
gas-liquid chromatography (GLC), where the stationary phase is a non-volatile liquid (Feng
et al., 2019).

OBJECTIVE
1. To analyse the fatty acids in corn oil and palm oil by using gas chromatography.
2. To study the principle of gas chromatography.
3.
RESULT

1. Record of retention time and peak area data

Table 1: Retention time of standard fatty acids

Standard / Sample Retention time (tR)

tR 1 tR 2 tR 3 tR average
Palmitic acid 2.238 2.229 2.246 2.238
Stearic acid 3.475 3.505 3.537 3.506
Palm oil Palmitic acid 2.258 2.254 2.256
Stearic acid 3.522 3.524 3.523
Corn oil Palmitic acid 2.262 2.232 2.247
Stearic acid 3.631 3.595 3.613

2. Peak Area data of standard fatty acids

Table 2: Peak Area of standard fatty acids

Standard / Sample Peak Area (cm2)

PA 1 PA 2 PA 3 PA average
Palmitic acid 2135.6 1842.4 1214.9 1730.97
Stearic acid 824.4 704.6 1302.1 943.70
Palm oil Palmitic acid 1458.5 1425.7 1442.10
Stearic acid 2240.5 2019.6 2130.05
Corn oil Palmitic acid 886.5 367.7 627.10
Stearic acid 6010.4 2173.3 4091.85
 CALCULATION

a. Response factor for each of the standard fatty acids

Response factor (Rf) = Concentration / Peak Area

Palmitic acid
8000 ppm
(Rf) =
1730.97
(Rf) = 4.6217

Stearic acid
8000 ppm
(Rf) =
943.70
(Rf) = 8.4773

b. Concentration of fatty acid in the sample

Concentration = Response factor (Rf) x Peak Area

Palm oil Corn oil

Palmitic Acid Stearic Acid Palmitic Acid Stearic Acid
Concentration of Concentration of Concentration of Concentration of
palmitic acid stearic acid palmitic acid stearic acid
= 4.6217 x 1442.10 = 8.4773 x 2130.05 = 4.6217 x 627.10 = 8.4773 x 4091.85
= 6664.95 ppm = 18057.07 ppm = 2898.27 ppm = 34687.84 ppm
DISCUSSION
The purpose of this experiment was to analyse the concentration of palmitic acid and
stearic acid in two different types of cooking oils by using gas chromatography. Samples of
two different types of cooking oil were used which comprise of palm oil and corn oil. The
known concentration of standard samples was prepared and then injected into the GC
injector port. Once the sample solution is injected into the instrument, it passes through the
gas stream that moves the samples into a column to allow the components to be separated
and eluted from the column outlet (Feng et al., 2019). A sequence of peaks is created in the
chromatogram by the separated substances flowing into it as they exit the column opening.
Data on the retention period and peak area of palmitic acid and stearic acid was recorded
and it is tabulated on Table 1 and Table 2.

According to the data shown in Table 1, we obtained the average of retention time
(tR) of palmitic acid was 2.238 minutes. Meanwhile for stearic acid, the average of retention
time (tR) was 3.506 minutes. Also, the retention time for palm oil and corn oil were measured
to demonstrate how long for both fatty acids maintained in the sample. We found that in
palm oil, the retention time for palmitic acid was 2.256 minutes and stearic acid was 3.523
minutes. Meanwhile for corn oil, the retention time of palmitic acid was 2.247 minutes and
stearic acid was 3.613 minutes respectively. Clark (2023) states that retention time is the
duration it takes for a certain component to pass through the column and reach the detector.
This duration is measured starting from the moment the sample is injected until the
compound's maximum peak height was displayed on the monitor. From the data above, we
could draw a conclusion that stearic acid has longer retention time in palm oil and corn oil.
This is due to the fact that stearic acid has longer carbon chain, C18 compared to palmitic
acid that have 16 carbon atoms long (Arnarson, 2019).

Data in Table 2 shows the peak area of standard fatty acids. For instance, the peak
area for palmitic acid was 1730.97 cm2 while stearic acid was 943.70 cm2. The peak area of
palmitic acid and stearic acid in palm oil were reported to be 1442.10 cm 2 and 2130.05 cm2.
Meanwhile, for corn oil it is obtained that the peak area of palmitic acid is 627.10 cm 2 and
stearic acid was 4091.85 cm2. According to Kaur (2020), the peak area indicates how many
components are present in a sample. Therefore, we could identify that stearic acid have the
highest amount in both palm oil and corn oil.

Each response factor (Rf) of standard sample fatty acid can be computed by dividing
the concentration of standard to the peak area of the standard. We calculated the response
factor for both palmitic acid and stearic acid was 4.6217 and 8.4473, respectively. The
concentration of standard sample fatty acid may be determined using this response factor
which then multiplied by peak area. Hence, we obtained that the concentration of palmitic
acid in palm oil was 6664.95 ppm and for stearic acid in palm oil was 18057.07 ppm. This
indicates that the concentration of stearic acid is higher compared to palmitic acid in palm oil.
Meanwhile the concentration of palmitic acid and stearic acid in corn oil were 2898.27 ppm
and 34687.84 ppm, respectively. The retention time of fatty acids may be compared with this
result. Hence, this explained why stearic acid eluted later in gas chromatograph.
Compounds with shorter retention times evaporate more quickly, whereas those with longer
retention times evaporate longer (Drawell, 2023).

Additionally, various safety precautions must be taken into account when doing this
experiment to prevent from obtaining false result. Firstly, make sure the second sample is
injected at least 10 minutes after the previous oil sample. This will ensure that that the
substance has fully eluted before the test. Moreover, the thumb must be placed above the
syringe plunger while injecting a sample. This is done to avoid sample blowback caused by
carrier gas pressure in the injection port. Lastly, gently tap the chromatographic syringe to
release any trapped air and eliminate any air bubbles.

CONCLUSION

In a nutshell, the retention time and peak area of each sample have been identified
by using gas chromatography. Gas chromatography is most frequently used methods in
determining fatty acids because it is precise and accurate. From the data, it is obtained that
palmitic acid has a shorter retention time compared to stearic acid because it evaporates
faster. Moreover, the response factor for both palmitic acid and stearic acid were 4.6217 and
8.4773, correspondingly. Also, we can conclude that stearic acid has the highest
concentration compared to palmitic acid in palm oil whereas for the corn oil, stearic acid was
thought to have higher concentration than palmitic acid. Overall, the objective of this
experiment was accomplished satisfactorily.
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Arnarson, A. (2019). 10 types of saturated fat reviewed. Healthline.

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Feng, T., Sun, M., Song, S., Zhuang, H., & Yao, L. (2019). Gas chromatography for
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Kaur, G. (2020). Gas Chromatography – A Brief Review. International Journal Of

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Libretexts. (2021). 17.1: Fatty acids. Chemistry LibreTexts.

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