ASSIGNMENT 2

FACULTY OF APPLIED SCIENCE

DIPLOMA IN SCIENCE

CHM 260: BASIC INSTRUMENT AND ANALYSIS

ASSIGNMENT 2

QUALITATIVE AND QUANTITATIVE ANALYSIS OF OLIEN IN

COOKING OILS BY GAS CHROMATOGRAPHY (GC)

NAME : CHARLES ROLEND RICHARD

MATRIX NUMBER : 2013219228

GROUP : AS1206A2

LECTURER’S NAME : JULENAH AG. NUDDIN

DATE OF SUBMISSION : 20th MARCH 2016

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1.0 INTRODUCTION

Quantitative analysis is the method that enables to determine the amount of a chemical in
a sample. The qualitative analysis is the identification of the constituents of chemical
elements and functional groups that present in the substance.

Olein which is also called as glyceryl trioleate, triolen is a colourless to yellowish, oily,
water-insoluble liquid, C57H104O6. Palm olien is a natural source of food with an equal
proportion of saturated and unsaturated fatty acids. It is also a source of Vitamin E and its
Omega 6 and Omega 9 which can helps to elevate normal blood cholesterol levels. Palm
olien is the liquid fraction by fractionation of palm oil after crystallization at controlled
temperatures. The physical characteristics of palm olien differ from those of palm oil. It is
fully liquid warm climate and has a narrow range of glycerides. The oils in common use for
making such products are mostly derived from seeds, and include soy bean oil, sun flower oil,
olive oil and others. Highly unsaturated oils generally have poor flavour and oxidative
stability, also often partially hydrogenated to overcome these defects.

Gas chromatography (GC) is a term to describe the group of analytical separation

techniques used to analyse volatile substances in the gas phase, whereas it relies on variance
in separating performance between a flowing mobile phase and a stationary phase to separate
the component in a mixture. The sample is carried by a moving stream through a tube packed
with a divided solid or may be covered with a film of a liquid (Nestola, 2015). The mobile
phase is a chemically inert gas that serves to carry the molecules of analyse through the
heated column. Meanwhile, the stationary phase consists of a packed column where the
packing or solid support itself acts as stationary phase, or is coated with the liquid stationary
phase. Most analytical gas chromatographs use capillary columns, where the stationary phase
coats the walls of a small-diameter tube directly. According to Nikolaeva (2014), the
separation of compounds is based on the different strengths of interaction of the compounds
with the stationary phase. The stronger the interaction is, the longer the compound interacts
with the stationary phase, and the more time it takes to migrate through the column, whereas
the interaction may be takes a long retention time. GC is known as one of the most crucial
tools in chemistry field because of its sensitivity and effectiveness in separating components
of mixture.
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Figure 1: Basic Gas Chromatography (GC) instruments

2.0 REVIEW OF LITERATURE

2.1 Olien
A study on the fatty acids composition of four different vegetable oils such as
red palm oil (RPO), palm olien (PO), corn oil (CO) and coconut oil (COC) by gas
chromatography was carried out. Eqbal (2011) stated that four different vegetable
oils were analysed for fatty acid concentration by gas chromatography. The results
shows that the predominant component of RPO and PO was oleic acid (18:0)
(44.616% and 49.482%) and palmitic acid (16:0) (42.465% and 36.768%)
respectively whilst the CO were rich in linoleic acid (18:0) with percentage of
47.189% but COC were rich in lauric acid (12:0) 46.458% compared to other oil
samples. Red palm olien and palm olein contains a healthy mixture of a fatty acid
composition, which is a healthy mixture of all the types of saturated and
unsaturated fatty acids.

2.2 Gas Chromatography

In early 1900s, Gas Chromatography (GC) was discovered by Mikhail
Semenovich Tsvett as a separation technique to separate compounds. In organic
chemistry, liquid-solid column chromatography is often used to separate organic
compounds in solutions. Among the various types of gas chromatography, gas-
liquid chromatography is the most common method that used to separate organic
compounds. The combination of gas chromatography and mass chromatograph
consists of an injection part, a column gas flow control equipment, ovens and
heaters for maintaining temperatures of the injection port and the column, an
integrator chart recorder and a detector.
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Analysis using gas-liquid chromatography (GLC) has become an important

tool for the analysis of emulsifiers. The development of auto-sampling technique
has made GLC more reproducible, which is necessary for quantitative analysis. A
new generation of gas chromatographs with computer connection enable the
calculation of emulsifier concentration become easier and faster. For more
complicated separations of components in emulsifiers, GC capillary columns can
be used with good result (Eka N., et al, 2013).

3.0 OBJECTIVE
i) To prepare the Fatty Acid Methyl Esterification. (FAMEs)
ii) To determine the retention time of olein.
iii) To evaluate the fatty acids composition of palm olein by gas chromatography.

4.0 METHODOLOGY
Instrument Gas Chromatography (C:/GCsolution/Data?Oleokimia/FAM.gcm) was
equipped with flame ionization detector and capillary column (30 m x 0.25 mm film).

Preparation of fatty acid methyl ester (FAME) was carried out according to Jumat et al.
(2006) and Siew et al. (1995). 1 mL of hexane was put into 0.1 mL vegetable oil and 1mL
sodium methoide (1.55g of MaOH in 50 mL of methanol) solution was added in the oil
solution. The solution stirred vigorously using vortex stirrer for 10 seconds. The solution was
left 10 minutes to separate out the clear solution of fatty acid methyl ester from the cloudy
aqueous layer. The upper layer was collected carefully. The fatty acid composition of four
different vegetable oils was determined using its fatty acid methyl esters and was injected
into gas-chromatography for analysis. The identification of the peaks was carried out by
retention times.

Sample Handling

The syringe will be rinsed before filling it with the sample. The volume of the sample may be
more than the required volume. No air bubbles in the syringe will be made sure. To remove
any air bubbles, the syringe may tap gently.
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The syringe will be held, needle up, and pushed the plunger to the required volume at eye
level. The excess sample will be removed using a tissue.

Experimental

First of all, the instrument is switched on. Then, the GC is set using the following condition,
initial oven temperature (70°C), Final oven temperature (70°C), Injection temperature
(180°C), and Detector temperature (180°C). Then, the component sample is injected and the
retention time of each component is determined individually. After that, the standard mixture
is injected and each component present is identified by comparing the retention time of each
component with the retention time of each single component determined previously. Then,
the unknown sample is injected and the component present in the unknown is identified.
Next, the column temperature is changes as follows, initial oven temperature (100°C), final
oven temperature (100°C), then the standard mixture is injected and comment on the effect of
reducing the temperature on the retention time and Rs of the components. After that, the oven
temperature is again change as follows, initial oven temperature (140°C) and final oven
temperature (140 C) The standard mixture is injected and comment on the effect of
increasing the temperature on the retention time and Rs of the components.
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Operation of GC
i) Instrument: Agilent 6890
The sample is injected into the septa for manual injection and the oven column
is checked to ensure the instrument is in the off mode. The instrument is
switched on, the warning tone is rang if the gas is not enough.
“INSTRUMENT1 ONLINE” is clicked in window screen and the instrument is
allowed to setup itself. The “METHOD”, “EDIT” and “OK” are clicked after
the setup is completed. The title is entered and “OK” is clicked. “MANUAL” is
clicked to select the injection and “BACK” is clicked to select injection
location. The information is kept. “SPLIT” and “SPLITLESS” mode are chosen
if the sample in either high concentration or low concentration. “COLUMN NO.
2”, “BACK”, “CONSTANT PRESSURE” and “”AMBIENT” are chosen at
column section, inlet column, and mode column and outlet column respectively.
Flame-ionization detector is used in detector column and parameter is injected
and “OK” is clicked. “APPLY”, “OK”, “EDIT INTEGRATION EVENT”,
“OK”, “SPECIFY REPORT”, “OK”, “RUN TIME CHECKLIST” and “OK” are
clicked and “SAVE” is clicked in the “FILE” to keep in command. Make sure
there are no bubbles are appeared in the syringe during injection. Go to “RUN
CONTROL”, “SAMPLE INFO” is clicked and “OPERATOR NAME” is set up
in column. Signal 2 is set and sample is filled. “RUN METHOD” is clicked and
the instrument is waited for injection. The sample is injected and “START”
button is pressed quickly on the instrument is waited for the injection. The
sample is injected and “START” button is pressed quickly on the instrument
then the syringe is pulled out from septa. “FILE” followed by “LOAD
SIGNAL” are clicked in data analysis. “CALIBRATION” and “NEW
CALIBRATION TABLE” is clicked, name is entered and the document is
printed. For the next sample, “DATA ANALYSIS” is clicked and the steps is
repeated.

5.0 EXPECTED OUTCOME

The fatty acid composition of four different vegetable oils was determined by capillary
GC as reported in table 1
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Name of fatty Shorthand Area % in Area % in Area %in Area % in

acid RPO PO CO COC
Caprylic 8:0 0.034 0.061 0.167 0.601
Capric 10:0 - - - 5.071
Lauric 12:0 0.173 0.230 0.042 46.458
Myristic 14:0 0.961 0.849 - 20.572
Palmtic 16:0 42.465 36.768 12.427 9.161
Stearic 18:0 0.395 - 11.442 2.936
Oleic 18:1 44.616 49.482 36.994 7.211
Linoleic 18:2 10.372 11.745 47.189 1.648
Linolenic 18:3 0.257 0.539 1.312 -
Arachidic 20:0 0.356 0.161 0.298 -
Heneicosanoi 21:0 - - - -
c
Beheric 22:0 0.059 0.061 0.151 -
Tricosanoic 23:0 0.022 0.031 - -
Lignoceric 24:0 0.067 0.066 - -

Table 1: Fatty acid composition of four different vegetable oils by Gas

Chromatography

Keyboard: Red Palm Oil (RPO), Palm Olien (PO), Corn Oil (CO) and Coconut Oil (COC)
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Figure 2: Retention time of olien

6.0 CLOSING REMARKS

Gas chromatography (GC) method was chosen to determine the qualitative and
quantitative analysis of olien in cooking oil as it is currently one of the most popular
methods for separating and analysing compounds. GC can be applied to the separation of
any compound that is either naturally volatile or can be converted to a volatile derivative.
This make GC is useful in separation of a number of small organic and inorganic
compounds Mass Spectrum (Serban, 2014). Furthermore, GC also provides separation
with excellent resolution of the compounds and it is suitable for quantitation when
standards are available. The area counts in the chromatogram can be measured and
expressed as normalised area counts reported to the peak area of an internal standard.
Apart from that, the fatty acid composition of vegetable oils was determined using its
fatty acid methyl esters and was injected into gas-chromatography for analysis. The
identification of the peaks was carried out by retention time. The high temperature and
high flow rates decrease the retention time, but also deteriorate the quality of separation.
The fatty acid composition of palm olein contain a healthy mixture of all types of fatty
acids saturated and unsaturated fatty acids. The fatty acid profile plays an important role
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to the physical-chemical properties. Hence, this is useful knowledge base that can be used
for advanced research in the future.

7.0 REFERENCE
Nestola, M., Friedrich, M., & Rainer, B. (2015). Universal Route to Aromatic
Hydrocarbon Analysis in Foodstuff: Gas Chromatography. American Chemical
Society, 87(12), 6195.

Nikolaeva, O. A., Patrushev, Y. V., & Sidelnikov, V. N. (2014). Programming of

Temperature and Flow Rate of Carrier Gas for Gas-Liquid Chromatograhpy.
Pleiades Publishing, 69(6), 559-562.

Eqbal M.A. D., Halimah A. S., Aminah A., & Zalifah M.K. (2011) Fatty Acids
Composition of Four Different Vegetable Oils (Red PLam Olien, Palm Olien, Corn
Oil and Coconut Oil) by Gas Chromatography. 2nd International Conference on
Chemistry on Chemistry and Chemical Engineering (Vol.14, pp. 31-34).

Eka N., Abdul, R., & Yaakob B.C.M (2013). Analysis of Emulsifier in Food Using
Chromatographic Technique. Journal of Food and Pharmaceutical Sciences.

Serban M. (2014). The Utilization of gas Chromatography/Mass Spectrometry in the

Profiling of Several Antioxidants in Botanicals. Retrieved September 4, 2015 from
http;//www.intechopen.com/books/advances-in-gas-chromatography/the-utilization-
of-gas-chromatography-mass-spectrometry-in-the-profiling-of-sveral-antioxidants-
in.html