International Journal of Food Properties

ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage: https://www.tandfonline.com/loi/ljfp20

Quality profile determination of palm olein:

potential markers for the detection of recycled
cooking oils

Yih Phing Khor, Biow Ing Sim, Faridah Abas, Oi Ming Lai, Yong Wang,
Yonghua Wang & Chin Ping Tan

To cite this article: Yih Phing Khor, Biow Ing Sim, Faridah Abas, Oi Ming Lai, Yong Wang,
Yonghua Wang & Chin Ping Tan (2019) Quality profile determination of palm olein: potential
markers for the detection of recycled cooking oils, International Journal of Food Properties, 22:1,
1172-1182, DOI: 10.1080/10942912.2019.1634098

To link to this article: https://doi.org/10.1080/10942912.2019.1634098

© 2019 Yih Phing Khor, Biow Ing Sim,

Faridah Abas, Oi Ming Lai, Yong Wang,
Yonghua Wang and Chin Ping Tan. Published
with license by Taylor & Francis Group, LLC.

Published online: 24 Jun 2019.

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INTERNATIONAL JOURNAL OF FOOD PROPERTIES
2019, VOL. 22, NO. 1, 1172–1182
https://doi.org/10.1080/10942912.2019.1634098

Quality profile determination of palm olein: potential markers for

the detection of recycled cooking oils
Yih Phing Khora, Biow Ing Sima, Faridah Abasb, Oi Ming Laic, Yong Wangd, Yonghua Wange,
and Chin Ping Tana,d,e
a
Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang,
Malaysia; bDepartment of Food Sciences, Faculty of Food Science and Technology, Universiti Putra Malaysia,
Serdang, Malaysia; cDepartment of Bioprocess Technology, Faculty of Biotechnology and Molecular Sciences,
Universiti Putra Malaysia, Serdang, Malaysia; dJNU-UPM International Joint Laboratory on Plant Oil Processing and
Safety (POPS), Department of Food Science and Engineering, Jinan University, Guangzhou, China; eGuangdong
Research Center of Lipid Science Applied Engineering Technology, School of Food Science and Engineering, South
China University of Technology, Guangzhou, China

ABSTRACT ARTICLE HISTORY

In this study, the safety and quality of commercial cooking oils were Received 4 March 2019
evaluated. The emphasis of this evaluation was on the presence of oxida- Revised 31 May 2019
tion and polymerization products in fresh oils, although the analyses were Accepted 14 June 2019
often conducted on used frying fats and oils. This was because polymerized KEYWORDS
triacylglycerols (PTGs) and monomeric oxidized triacylglycerols (oxTAGs) Polymerized triacylglycerol;
have been proposed as potential indicators of the adulteration of palm monomeric oxidized
olein. The oil quality was evaluated based on PTG content, the presence of triacylglycerol; total polar
epoxy, keto, and hydroxy acids, fatty acid composition, and smoke point. compound; fatty acid
Principal component analysis (PCA) was conducted to identify the relation- composition; smoke point;
ships among the analytical parameters. The total polar compound content oil safety
of all fresh oil samples was within the safety limit for human consumption
(< 25% polar compounds). TAG oligomers or epoxy, keto, or hydroxy acids
were not detected in any of the fresh oil samples. Most of the packet oils
had lower smoke point (< 200 °C) and linoleic acid content than the bottled
oils. The pure palm olein samples were found to be better in terms of
overall oil quality, as indicated by the PCA biplots of all analytical
parameters.
Abbreviations: ANOVA: analysis of variance; BPO: blended palm olein; DAG:
diacylglycerol; FAME: fatty acid methyl ester; FFA: free fatty acid; HPLC:
high-performance liquid chromatography; HPSEC: high-performance size
exclusion chromatography; PC: principal component; PO: packet pure
palm olein; PPO: pure palm olein; PCA: principal component analysis; SPE:
solid-phase extraction; TAG: triacylglycerol; TPC: total polar compound

Introduction
Polymerized triacylglycerols, which are of high molecular weight, are normally abundantly formed in
cooking oil during heat treatment. These byproducts are normally nonvolatile, more polar than the
unaltered triacylglycerols, and their presence reduces the nutritional value of the oil.[1,2] The
detection of the isolated polar compound and its constituents by high-performance size-exclusion
chromatography provides a comprehensive evaluation of the impact of the three common degrada-
tion pathways that occur in oils during heat treatment, namely, polymerization, oxidation, and

CONTACT Chin Ping Tan tancp@upm.edu.my Department of Food Technology, Faculty of Food Science and Technology,
Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/ljfp.
© 2019 Yih Phing Khor, Biow Ing Sim, Faridah Abas, Oi Ming Lai, Yong Wang, Yonghua Wang and Chin Ping Tan. Published with license by Taylor &
Francis Group, LLC.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 1173

hydrolysis.[3,4] Therefore, it is vital to evaluate the quality of cooking oils, even fresh ones, as the
presence of these compounds will subsequently affect the quality of fried food products.
Polar fatty acids such as epoxy, keto, and hydroxy acids are relatively stable final products that are
formed from the decomposition of hydroperoxides.[5] These oxygenated compounds are reported to
be present at high levels, especially in thermally oxidized foods.[6] These oxidized fatty acids,
particularly long-chain epoxy fatty acids that can act as protoxins, are claimed to have toxic or
adverse effects. This term ‘protoxin’ indicates that a compound only becomes a toxin when it is
altered during metabolism.[7] Therefore, the safety issues related to the presence of these oxygenated
fatty acids in the oil cannot be underestimated.
Polymerized triacylglycerols and monomeric oxidized triacylglycerols have been proposed as
potential indicators of the adulteration of palm olein. Because their molecular weights are generally
higher, they are more difficult to remove during the refining process. The aim of this work was to
determine the presence of oxidative polymerization products in fresh commercial cooking oils,
although these analyses were often conducted on used frying fats and oils. This was considered
part of the safety evaluation of commercial cooking oils available in Malaysia, as these compounds
should not be present in fresh oil. Besides, the relationships between the analytical parameters were
studied and the quality of commercial cooking oil samples from different categories, for instance,
bottled pure palm olein, blended palm olein, and packet pure palm olein were evaluated and
compared. The safety parameters of commercial cooking oils must be closely monitored because
these oils are widely used for culinary purposes, e.g., in deep frying, cooking, and baking.

Materials and methods

Materials
A total of 23 commercial cooking oil samples were acquired from a local supermarket (Selangor,
Malaysia); the samples were categorized into three main groups, namely, pure palm olein, blended
palm olein, and packet pure palm olein. Pure palm olein is denoted PPO1 to PPO9, blended palm
olein is denoted BPO1 to BPO3, and packet pure palm olein is denoted PO1 to PO11. The blended
palm olein is a mixture consisting of major percentage of palm olein and minimal amount of several
vegetable oils such as soybean, sunflower, sesame, and peanut oils.

Chemicals
All solvents and chemicals used in the analyses were of analytical grade. For column chromatogra-
phy, silica gel 60 (0.063–0.200 mm) was purchased from Merck (Darmstadt, Germany).
Tetrahydrofuran (HPLC grade), which was used as the mobile phase for high-performance size-
exclusion chromatography (HPSEC), and platinum (IV) oxide hydrate were acquired from Thermo
Fisher Scientific (Waltham, Massachusetts, USA). Analytical standards for the analysis of monomeric
oxidized triacylglycerols such as methyl heneicosanoate, methyl 12-hydroxystearate, and methyl 12-
oxostearate were purchased from Sigma-Aldrich (Darmstadt, Germany), while methyl trans-9,10-
epoxystearate was purchased from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA). The Supelco
37 component FAME mix and methyl octanoate used in fatty acid composition analysis were
purchased from Sigma-Aldrich (Darmstadt, Germany).

Physical property
Determination of smoke point: The smoke point of all commercial cooking oil samples was determined
according to the official analytical method (Cc 91-48) of the American Oil Chemists’ Society.[8]
1174 Y. P. KHOR ET AL.

Chemical properties
Total polar compounds and their distribution: An amount of 0.5 g commercial cooking oil samples
were separated into two fractions, a polar fraction and a nonpolar fraction, by conventional silica gel
column chromatography. The separation method was conducted according to a group of European
researchers but with slight modifications.[6] A total of 10 g of silica was adjusted to a moisture
content of 5% (w/w) and was placed in a glass column (40 cm × 1 cm I.D.). First, 75 mL of n-hexane-
diethyl ether (90:10, v/v) was used to elute the unaltered TAGs (the nonpolar fraction). Our interest
in this study was the polar fraction, which was eluted with 75 mL of diethyl ether, a solvent of higher
polarity than the previous eluent. The polar fraction collected in diethyl ether was evaporated under
reduced pressure using a rotary evaporator at 40 °C for 10 min. After evaporation, the round-bottom
flask containing the residue was placed in an oven at 105 °C to ensure that the residual solvent was
fully evaporated. The collected polar fraction was then accurately weighed and dissolved in tetra-
hydrofuran (THF) at a concentration of 50 mg/ml. The sample was filtered and analyzed by high-
performance size-exclusion chromatography using a refractive index detector (HPLC-SEC/RI).
High-performance size-exclusion chromatography (HPSEC): The distribution of total polar
compounds in the commercial cooking oils was analyzed using HPSEC. TAG oligomers, TAG
dimers, monomeric oxidized triacylglycerols, DAG, and FFA were quantitated using a Shimadzu
HPLC equipped with a SIL-10AD injector, an LC-20AD pump, and a RID-10A refractive index
detector. Two 100-Å and 500-Å phenogel columns (30 cm × 0.78 cm I.D., film thickness 5 µm) with
porous, highly cross-linked styrene-divinyl benzene copolymers (Phenomenex, Torrance, CA, USA)
as the stationary phase were connected in series to improve the peak resolution. Tetrahydrofuran
was used as the mobile phase, and the flow rate was set to 1 mL/min. Each group of the polar
fraction distributions was quantified using the following equations:
 X 
Poligomer ¼ Aoligomer = A PTPC
 X 
Pdimer ¼ Adimer = A PTPC
 X 
PoxTAG ¼ AoxTAG = A PTPC
 X 
PDAG ¼ ADAG = A PTPC
 X 
PFFA ¼ AFFA = A PTPC

where PTPC, Poligomer, Pdimer, PoxTAG, PDAG, and PFFA represent the percentages of total polar
compound, TAG oligomers, TAG dimers, monomeric oxidized triacylglycerols, diacylglycerols,
and free fatty acids found in the polar fraction of the oil sample respectively; Aoligomer, Adimer,
AoxTAG, ADAG, and AFFA represent the peak area of each specific fractions; and ∑A represents the
sum of all peak areas.[6]
Epoxy, keto, and hydroxy fatty acids: The commercial cooking oil samples were derivatized with
sodium methoxide to produce fatty acid methyl esters (FAMEs), which are more volatile and more
suitable for GC analysis. A 300-mg oil sample was weighed followed by the addition of 3 mL of tert-
butyl methyl ether (TBME) and 1.5 mL of 0.2 M sodium methoxide in methanol. The mixture was
vortexed for 1 min and allowed to stand at room temperature for 2 min. For the neutralization step,
0.1 mL of 0.5 M sulfuric acid was added, and the mixture was vortexed for 5 s. Subsequently, 3 mL of
ultrapure water was added, and the mixture was vortexed for 10 s and centrifuged for 5 min at
4000 rpm. After centrifugation, the supernatant was transferred to a separate test tube and evapo-
rated to dryness under nitrogen.[6]
Solid-phase extraction (SPE): SPE is an important sample pretreatment step that is used to
fractionate FAMEs into polar and nonpolar fractions using eluents with different polarities. A silica-
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 1175

based SPE cartridge containing 1 g sorbent (Phenomenex, Torrance, CA, USA) was used in the
analysis. After preconditioning, 100 mg of FAMEs dissolved in 2 mL of n-hexane-diethyl ether (98:2;
v/v) were transferred into the SPE cartridge. Fifteen milliliters of n-hexane-diethyl ether (98:2; v/v)
was used to elute the nonpolar fraction. The separation was followed by elution of the polar fraction
using 25 mL of diethyl ether. A total of 1 mL of methyl heneicosanoate (500 µg/mL) dissolved in
TBME solution was added to the polar fraction as an internal standard. The mixture was vortexed,
and the solvent was evaporated to dryness under nitrogen.[6]
Hydrogenation: A total of 2 mL of methanol and platinum (IV) oxide hydrate, which acted as the
metal catalyst for the hydrogenation process, was added to the collected polar FAMEs. The hydro-
genation process was conducted at room temperature by bubbling hydrogen gas into the sample for
10 min. After hydrogenation, the methanol was evaporated under nitrogen. The sample was
dissolved in 1.5 mL of diethyl ether, filtered, and injected into a gas chromatograph equipped with
a flame ionization detector (GC-FID).[6]
Gas chromatography: An Agilent 6890 series chromatography system (Agilent Technologies,
Santa Clara, CA, USA) equipped with a split-splitless injector was used to analyze the epoxy, keto,
and hydroxy acids. The injector was operated at 250 °C, and the split ratio was set to 20:1. The
separation was performed using a J&W DB-Wax fused-silica capillary column (60 m × 0.25 mm
internal diameter and a film thickness of 0.25 µm) (J&W Scientific, Folsom, CA, USA). The flame
ionization detector temperature was set to 250 °C. The detector flow rates for hydrogen, air, and
nitrogen were 40 mL/min, 450 mL/min, and 45 mL/min, respectively. The column flow rate of
nitrogen as the carrier gas was 1 mL/min. The analysis was performed under isothermal conditions
at 240 °C for 30 min.[6] The signal-to-noise ratios of each peaks were monitored to ensure the
reliability of the quantified data. The monomeric oxidized triacylglycerols contents were determined
from the calibration curve constructed by plotting the concentration of standards versus the
corresponding peak areas.
Fatty acid composition: A base-catalyzed method was used to derivatize the fatty acids at room
temperature. A 100-mg sample of each commercial cooking oil was accurately weighed. Each sample
was dissolved in 2 mL of hexane containing 1 mg/mL methyl tridecanoate (C13:0), followed by the
addition of 0.1 mL of 2 M potassium hydroxide in methanol. The mixtures were vortexed for 15 s
and centrifuged at 4000 rpm for 5 min. The organic layers were filtered and analyzed using a gas
chromatograph equipped with a flame ionization detector (GC-FID).[9]
Gas chromatography: An Agilent 6890 Series chromatography system (Agilent Technologies,
Santa Clara, CA, USA) equipped with a split-splitless injector was used to analyze the FAMEs.
The split mode was set at 20:1, the split ratio was set at 250 °C, and the flame ionization detector was
set at 280 °C. An SGE BPX70 column (25 m × 0.32 mm internal diameter and 0.25 µm film
thickness) was used in this analysis. The hydrogen gas flow was 40 mL/min, the air flow was 450 mL/
min, and the flow of nitrogen, which acted as the auxiliary gas, was 25 mL/min. The carrier gas,
which was nitrogen, was fixed at 10 psi. The oven temperature program was as follows: hold at 100 °
C for 5 min, ramp to 240 °C at 4 °C/min, and hold for 20 min. A calibration curve for caprylic acid
(R2 = 0.99) was constructed separately to quantify its concentration.

Statistical analysis
All the measurement data were analyzed using one-way ANOVA, and p-values < 0.05 were
considered significant. Data analyses were conducted to identify any significant differences in the
collected data and were performed using Minitab software (Minitab Version 17.1, Minitab Pty Ltd,
Sydney, NSW, Australia). Principal component analysis (PCA) was also performed to study the
relationships among analytical parameters and at the same time to evaluate the interaction between
analytical parameters and types of commercial cooking oil samples. The PCA evaluation was
performed using XLSTAT software (version 2018, Addinsoft, New York, USA).
1176 Y. P. KHOR ET AL.

Results and discussion

Smoke point
Smoke point is one of the important physical parameters that is used to gauge the stability of oils.
The smoke point is generally related to the accumulation of decomposition products. The generation
of smoke marks the beginning of both flavor and nutritional degradation of the oil.[10] A low smoke
point is generally related to low thermal stability.[11] In other words, a higher smoke point indicates
that the oil is of better quality and contains lower levels of FFA. The results presented in Table 1
show that most of the packet oils (pure palm olein) had lower smoke points (< 200°C) than the
bottled oils (pure palm olein and blended palm olein). Generally, these packet oils are sold at the
lowest retail prices and are considered low-cost products. The use of low-quality crude palm oil with
relatively high FFA content as the input for the refining process might explain the lower smoke point
of the refined packet oils compared to bottled pure palm olein and blended palm olein. In addition,
the process used to refine the packet oils might not be well monitored and controlled; refining is an
important step in removing extraneous impurities and volatile trace constituents such as free fatty
acids that reduce the smoke point of the oil.

Total polar compounds (TPC) and their distribution

Determination of the level of total polar compounds is the most reliable measure of fat deterioration
and quality control for fats and oils.[12] This is because it provides information on the total amount
of new compounds formed upon various heat treatments. This gravimetric method is precise and
accurate because it isolates polar compounds based on adsorption chromatography.[13] Table 2
summarizes the content of total polar compounds and their constituents such as triacylglygerol
(TAG) dimers, monomeric oxidized TAG, diacylglycerols (DAG), and free fatty acids in the oil
samples.

Table 1. Determination of the smoke points of commercial cooking oils.

Type of cooking oil Sample Smoke point (°C)
Pure Palm Olein (PPO) PPO1 221.00 ± 1.41i
PPO2 181.00 ± 1.41a
PPO3 189.50 ± 0.71cde
PPO4 200.00 ± 1.41fg
PPO5 226.50 ± 0.71j
PPO6 209.50 ± 0.71h
PPO7 197.50 ± 0.71f
PPO8 209.50 ± 0.71h
PPO9 222.50 ± 0.71ij
Blended Palm Olein (BPO) BPO1 225.00 ± 1.41ij
BPO2 203.50 ± 0.71g
BPO3 185.50 ± 0.71bc
Packet Oil (PO) PO1 184.70 ± 1.84ab
PO2 208.00 ± 1.41h
PO3 189.50 ± 0.71cde
PO4 192.00 ± 1.41e
PO5 189.50 ± 0.71cde
PO6 191.35 ± 0.92de
PO7 196.90 ± 0.14f
PO8 187.55 ± 0.64bcd
PO9 202.50 ± 0.71g
PO10 187.70 ± 0.42bcd
PO11 192.50 ± 0.71e
*Data are expressed as the mean ± standard deviation (n = 4). Mean values
with different superscripts in the same column are significantly different at
p < 0.05.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 1177

Table 2. Determination of the polar fraction content and composition of commercial cooking oils.
Polar fraction composition (%, g 100 g−1)
Type of Polar fraction content TAG Oxidized TAG Free Fatty
cooking oil Sample (%, g 100 g−1) Oligomers TAG Dimers Monomers Diacylglycerols Acids
Pure Palm PPO1 7.04 ± 0.13abc N.D. 0.31 ± 0.04bcdef 0.97 ± 0.11ab 5.58 ± 0.01ab 0.18 ± 0.01ab
Olein (PPO) PPO2 8.47 ± 0.06cd N.D. 0.39 ± 0.01ef 1.10 ± 0.24abc 6.40 ± 0.08abc 0.58 ± 0.10c
PPO3 7.52 ± 0.00abc N.D. 0.36 ± 0.07def 0.80 ± 0.09ab 6.01 ± 0.05ab 0.36 ± 0.21abc
PPO4 6.96 ± 0.91ab N.D. 0.28 ± 0.04bcdef 0.89 ± 0.08ab 5.56 ± 0.75ab 0.22 ± 0.03ab
PPO5 7.87 ± 0.30abc N.D. 0.24 ± 0.01abcde 1.18 ± 0.38abc 6.22 ± 0.21ab 0.22 ± 0.12ab
PPO6 7.26 ± 0.33abc N.D. 0.26 ± 0.02bcdef 0.94 ± 0.26ab 5.80 ± 0.12ab 0.25 ± 0.03ab
PPO7 8.19 ± 0.40abcd N.D. 0.29 ± 0.03bcdef 0.84 ± 0.06ab 6.73 ± 0.30abc 0.34 ± 0.07abc
PPO8 7.22 ± 0.17abc N.D. 0.42 ± 0.01f 1.00 ± 0.11ab 5.56 ± 0.17ab 0.24 ± 0.11ab
PPO9 8.25 ± 0.01abcd N.D. 0.38 ± 0.01ef 1.73 ± 0.16cd 5.99 ± 0.19ab 0.14 ± 0.00ab
Blended Palm BPO1 7.31 ± 0.01abc N.D. 0.24 ± 0.02abcde 0.93 ± 0.17ab 5.98 ± 0.08ab 0.16 ± 0.08ab
Olein (BPO) BPO2 7.40 ± 0.26abc N.D. 0.32 ± 0.03bcdef 1.30 ± 0.21abcd 5.47 ± 0.42a 0.30 ± 0.08abc
BPO3 9.67 ± 0.38d N.D. 0.32 ± 0.04bcdef 1.41 ± 0.32bcd 7.52 ± 0.06c 0.42 ± 0.03bc
Packet Oil (PO) PO1 7.67 ± 0.13abc N.D. 0.27 ± 0.06bcdef 0.93 ± 0.15ab 6.21 ± 0.25ab 0.25 ± 0.03ab
PO2 6.84 ± 0.02a N.D. 0.17 ± 0.01abc 0.66 ± 0.04a 5.80 ± 0.05ab 0.20 ± 0.03ab
PO3 7.91 ± 0.01abc N.D. 0.36 ± 0.00def 1.29 ± 0.01abcd 5.99 ± 0.02ab 0.28 ± 0.01ab
PO4 7.69 ± 0.11abc N.D. 0.29 ± 0.10bcdef 1.12 ± 0.01abc 6.10 ± 0.03ab 0.19 ± 0.01ab
PO5 8.24 ± 0.02abcd N.D. 0.16 ± 0.04ab 1.35 ± 0.02abcd 6.38 ± 0.02abc 0.34 ± 0.07abc
PO6 7.37 ± 0.11abc N.D. 0.19 ± 0.01abcd 0.79 ± 0.14ab 6.17 ± 0.09ab 0.21 ± 0.05ab
PO7 7.61 ± 0.38abc N.D. 0.34 ± 0.06cdef 1.05 ± 0.28abc 5.89 ± 0.19ab 0.32 ± 0.04abc
PO8 8.35 ± 1.00bcd N.D. 0.08 ± 0.01a 1.93 ± 0.13d 6.10 ± 0.90ab 0.24 ± 0.03ab
PO9 7.57 ± 0.59abc N.D. 0.16 ± 0.05ab 1.08 ± 0.11abc 6.03 ± 0.44ab 0.29 ± 0.00abc
PO10 8.18 ± 0.21abcd N.D. 0.24 ± 0.01abcde 1.15 ± 0.03abc 6.40 ± 0.14abc 0.39 ± 0.05abc
PO11 7.41 ± 0.37abc N.D. 0.76 ± 0.10g 0.87 ± 0.04ab 5.66 ± 0.28ab 0.12 ± 0.05a
N.D.: not detected
*Data are expressed as the mean ± standard deviation (n = 4). Mean values with different superscripts in the same column are
significantly different at p < 0.05.

Polymerized TAG, the major decomposition products of cooking oil, are nonvolatile and relatively
stable.[14] These compounds are separated based on their molecular weights using high-performance
size-exclusion chromatography; in this method, TAG oligomers of the highest molecular weights will be
eluted first, followed by TAG dimers, monomeric oxidized TAG, DAG, and, lastly, free fatty acids.
Because palm olein contains a high amount of DAG, the percentage of total polar compounds in palm
olein is slightly higher than that in other soft oils.[15] Therefore, determination of the distribution of polar
compounds is very important.
The total polar compounds for all fresh oil samples fell within the safety limit for human
consumption (< 25% polar compounds) set in many European countries.[16] No TAG oligomers
were detected in any of the commercial oil samples. The TPC content in PPO ranged from 6.96% to
8.47%, BPO ranged from 7.31% to 9.67%, and PO ranged from 6.84% to 8.35%. High TPC is
unfavorable because it accelerates the oil oxidation process, increases the oil’s viscosity, reduces heat
transfer, causes a foaming reaction, produces undesirable color in the fried food, and increases the
rate of oil absorption by foods.[14] The TAG dimer levels ranged from 0.08% to 0.76% in all fresh
commercial oil samples, which were far below the rejection limit (12–13%).[17]
The presence of TAG dimers in fresh cooking oils is normal. This is because dimerization of
triacylglycerols occurs during the deodorization step of the oil refining process due to the high-
temperature processing. Acyclic dimers of triacylglycerols, such as oxygenated dimers (C-O-C), and
nonpolar dimers (C-C bridges), can be formed when the oil is subjected to extreme conditions.[14] These
dimers could possibly lead to the formation of polyaromatic hydrocarbons or cyclic compounds.[18]
The fresh commercial cooking oils also possessed a relatively high amount of free fatty acids
(FFA) compared to the values reported in the literature.[19] In fact, FFA are volatile, and they are
mostly removed by steam during the deodorization process.[20] FFA, which possess both hydrophilic
and hydrophobic groups in the same molecule, act as pro-oxidants.[21] FFA normally remain at the
surface of cooking oils and decrease the surface tension of the cooking oil. Thus, they help increase
1178 Y. P. KHOR ET AL.

the rate of diffusion of oxygen from the headspace into the oil and thereby accelerate oil
oxidation.[20] Therefore, a good-quality crude palm oil with low FFA is a prerequisite for the
production of good-quality refined palm olein as well as for minimization of refining losses.
Refined oils with high FFA levels might also result from poor vacuum conditions during the
deodorization process, inadequate steam sparging, or leakage of air.

Epoxy, keto, and hydroxy fatty acids

Based on the results shown in Table 3, epoxy, keto, and hydroxy fatty acids were not detected in any
of the 23 commercial fresh cooking oil samples. The analysis of epoxy, keto, and hydroxy fatty acids
is normally conducted on thermally oxidized oils because they are reported to form in high amounts
during heat treatment. However, one study reported that epoxy fatty acids were detected in amounts
ranging from 0.03 to 2 mg g−1 even in fresh oil that was high in polyunsaturated fatty acids.[7]
According to literature, the major precursors of these oxygenated fatty acids are polyunsaturated
fatty acids.[7]. The formation of epoxy fatty acids was related to the concentration of unsaturated
fatty acids present in the oil and not to the relative oxidizability rates. Therefore, palm olein, which is
high in monounsaturated fatty acids (oleic acid), might also contribute to the formation of these
oxygenated fatty acids during heat treatment. These compounds are nonvolatile, relatively stable and
remain in the oil after their formation. Therefore, to ensure the standard quality and safety of the
commercial cooking oils, it is also important to evaluate whether or not the fresh commercial
cooking oils contain these oxygenated compounds.

Fatty acid composition

Determination of fatty acid composition is a pivotal issue in the field of food research, especially in lipid
studies.[22] Table 4 shows the fatty acid compositions of 23 commercial cooking oil samples available in

Table 3. Quantitative determination of epoxyFAMEs, ketoFAMEs and hydroxyFAMEs (g/100 g) in commercial cooking
oils.
Type of cooking oil Sample EpoxyFAMEs KetoFAMEs HydroxyFAMEs
Pure Palm Olein (PPO) PPO1 N.D. N.D. N.D.
PPO2 N.D. N.D. N.D.
PPO3 N.D. N.D. N.D.
PPO4 N.D. N.D. N.D.
PPO5 N.D. N.D. N.D.
PPO6 N.D. N.D. N.D.
PPO7 N.D. N.D. N.D.
PPO8 N.D. N.D. N.D.
PPO9 N.D. N.D. N.D.
Blended Palm Olein (BPO) BPO1 N.D. N.D. N.D.
BPO2 N.D. N.D. N.D.
BPO3 N.D. N.D. N.D.
Packet Oil (PO) PO1 N.D. N.D. N.D.
PO2 N.D. N.D. N.D.
PO3 N.D. N.D. N.D.
PO4 N.D. N.D. N.D.
PO5 N.D. N.D. N.D.
PO6 N.D. N.D. N.D.
PO7 N.D. N.D. N.D.
PO8 N.D. N.D. N.D.
PO9 N.D. N.D. N.D.
PO10 N.D. N.D. N.D.
PO11 N.D. N.D. N.D.
N.D.: not detected
*Data are expressed as the mean ± standard deviation (n = 4). Mean values with different superscripts in the same
column are significantly different at p < 0.05.
Table 4. Determination of the fatty acid composition of commercial cooking oils.
Fatty acid composition (relative percentages)
Type of cooking oil Sample C12:0 C14:0 C16:0 C18:0 C18:1 C18:2 C18:3 C18:2/C16:0 C8 (ppm)
Pure Palm Olein (PPO) PPO1 0.29 ± 0.00efgh 1.57 ± 0.32a 38.28 ± 0.07defg 4.18 ± 0.02ijk 44.70 ± 0.21bc 10.94 ± 0.01fghij 0.00 ± 0.00a 0.29 ± 0.00ij 0.15 ± 0.22a
PPO2 0.52 ± 0.00i 1.80 ± 0.01a 36.26 ± 0.02bc 3.99 ± 0.01cdefghi 45.04 ± 0.02bc 12.33 ± 0.00k 0.00 ± 0.00a 0.34 ± 0.00n 0.88 ± 0.01c
PPO3 0.25 ± 0.00bcdefg 1.82 ± 0.16a 38.88 ± 0.05g 4.22 ± 0.02jk 44.12 ± 0.06b 10.68 ± 0.03defgh 0.00 ± 0.00a 0.27 ± 0.00h 0.00 ± 0.00a
PPO4 0.32 ± 0.00fgh 1.79 ± 0.04a 36.55 ± 0.01bcde 3.86 ± 0.01bcd 45.90 ± 0.04bc 11.54 ± 0.00j 0.00 ± 0.00a 0.32 ± 0.00m 0.07 ± 0.03a
PPO5 0.36 ± 0.00gh 1.47 ± 0.01a 36.42 ± 0.01bcd 3.73 ± 0.01b 46.90 ± 0.00c 11.07 ± 0.01ghij 0.00 ± 0.00a 0.30 ± 0.00k 0.54 ± 0.12b
PPO6 0.39 ± 0.00h 1.86 ± 0.03a 37.97 ± 0.00cdefg 3.96 ± 0.00cdefgh 45.81 ± 0.02bc 9.97 ± 0.04bc 0.00 ± 0.00a 0.26 ± 0.00de 0.20 ± 0.02a
PPO7 0.12 ± 0.00a 1.81 ± 0.07a 38.50 ± 0.01efg 3.86 ± 0.00bc 44.54 ± 0.05b 11.17 ± 0.01hij 0.00 ± 0.00a 0.29 ± 0.00j 0.00 ± 0.00a
PPO8 0.32 ± 0.00fgh 1.80 ± 0.06a 39.02 ± 0.00g 4.26 ± 0.01k 43.96 ± 0.05b 10.61 ± 0.01cdefgh 0.00 ± 0.00a 0.27 ± 0.00gh 0.17 ± 0.02a
PPO9 0.29 ± 0.00defgh 1.49 ± 0.00a 39.17 ± 0.00g 4.09 ± 0.00fghijk 44.57 ± 0.01b 10.37 ± 0.00cdef 0.00 ± 0.00a 0.26 ± 0.00ef 0.00 ± 0.00a
Blended Palm Olein (BPO) BPO1 0.25 ± 0.00bcdefg 1.50 ± 0.02a 36.86 ± 0.00bcdef 4.09 ± 0.01fghijk 45.87 ± 0.01bc 11.41 ± 0.00ij 0.00 ± 0.00a 0.31 ± 0.00l 0.00 ± 0.00a
BPO2 0.31 ± 0.00fgh 2.08 ± 0.17a 37.98 ± 0.01cdefg 4.05 ± 0.01cdefghij 44.68 ± 0.11bc 10.80 ± 0.04efghi 0.00 ± 0.00a 0.28 ± 0.00i 2.13 ± 0.25d
BPO3 0.32 ± 0.00fgh 1.94 ± 0.10a 35.63 ± 0.00b 3.98 ± 0.01cdefgh 45.56 ± 0.07bc 12.53 ± 0.02k 0.00 ± 0.00a 0.35 ± 0.00° 0.12 ± 0.05a
Packet Oil (PO) PO1 0.13 ± 0.00ab 1.97 ± 0.01a 39.70 ± 0.01g 3.91 ± 0.00bcdef 44.13 ± 0.02b 10.16 ± 0.00bcde 0.00 ± 0.00a 0.26 ± 0.00bc 0.00 ± 0.00a
PO2 0.34 ± 0.00gh 2.11 ± 0.04a 37.93 ± 0.00cdefg 3.89 ± 0.00bcde 45.30 ± 0.03bc 10.39 ± 0.01cdef 0.00 ± 0.00a 0.27 ± 0.00h 0.00 ± 0.00a
PO3 0.19 ± 0.00abcde 2.22 ± 0.06a 39.10 ± 0.04g 4.12 ± 0.01hijk 44.30 ± 0.00b 10.07 ± 0.01bcd 0.00 ± 0.00a 0.26 ± 0.00bc 0.00 ± 0.00a
PO4 0.14 ± 0.00abc 2.79 ± 0.18a 39.34 ± 0.06g 3.92 ± 0.00bcdefg 43.63 ± 0.11b 10.18 ± 0.01bcde 0.00 ± 0.00a 0.26 ± 0.00cd 0.00 ± 0.00a
PO5 0.26 ± 0.00defg 2.34 ± 0.09a 38.52 ± 0.02efg 4.01 ± 0.01cdefghi 45.21 ± 0.06bc 9.63 ± 0.00b 0.00 ± 0.00a 0.25 ± 0.00a 0.00 ± 0.00a
PO6 0.22 ± 0.00abcdef 1.70 ± 0.04a 38.75 ± 0.03fg 4.07 ± 0.00efghijk 44.82 ± 0.02bc 10.42 ± 0.00cdefg 0.00 ± 0.00a 0.27 ± 0.00fg 0.00 ± 0.00a
PO7 0.17 ± 0.00abcd 2.50 ± 0.03a 38.95 ± 0.00g 4.11 ± 0.00ghijk 44.19 ± 0.02b 10.07 ± 0.00bcd 0.00 ± 0.00a 0.26 ± 0.00cd 0.00 ± 0.00a
PO8 0.10 ± 0.14a 19.15 ± 6.19 b 32.82 ± 2.35a 3.23 ± 0.22a 36.40 ± 2.71a 8.31 ± 0.77a 0.00 ± 0.00a 0.25 ± 0.01ab 0.00 ± 0.00a
PO9 0.28 ± 0.00defgh 2.23 ± 0.09a 38.84 ± 0.02fg 4.05 ± 0.00defghij 44.31 ± 0.05b 10.25 ± 0.01bcde 0.00 ± 0.00a 0.26 ± 0.00e 0.00 ± 0.02a
PO10 0.26 ± 0.00cdefg 1.96 ± 0.07a 37.82 ± 0.01cdefg 3.93 ± 0.00bcdefg 45.54 ± 0.05bc 10.46 ± 0.02cdefg 0.00 ± 0.00a 0.28 ± 0.00h 0.00 ± 0.01a
PO11 0.28 ± 0.00defgh 2.00 ± 0.02a 38.99 ± 0.03g 4.03 ± 0.00cdefghi 44.37 ± 0.04b 10.30 ± 0.02cdef 0.00 ± 0.00a 0.26 ± 0.00ef 0.00 ± 0.00a
*Data are expressed as the mean ± standard deviation (n = 4). Mean values with different superscripts in the same column are significantly different at p < 0.05.
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1180 Y. P. KHOR ET AL.

the Malaysian market. The results of the analysis of cooking oil packaged in plastic bottles were in
accordance with the findings of the Malaysian Palm Oil Board.[23] Sample BPO3 contained
a significantly high (p < 0.05) percentage of oleic and linoleic acid, likely because this sample was
blended with sesame oil and groundnut oil, both of which are high in oleic and linoleic acid. Most of the
packet oils showed significantly low (p < 0.05) linoleic acid content compared with the bottled oils.
There was also a possibility that the palm oil was not well fractionated, especially in the packet
oils, as some fat solids were noticeable in the liquid fraction despite the fact that the oil samples were
carefully maintained at ambient temperature. This might explain the higher content of myristic and
palmitic acid in packet oils compared with bottled oils; an exception was sample PO8, which had
significantly low (p < 0.05) percentages of palmitic, stearic, oleic, and linoleic acids but a significantly
high (p < 0.05) percentage of myristic acid.
Caprylic (C8:0) acid was quantified separately because it is commonly detected in palm kernel oil
but not in palm olein.[24] Short-chain fatty acids such as caprylic acid might be formed in small
amounts during the thermal decomposition of fatty acid hydroperoxides. One study showed that all
the major unsaturated fatty acids, including oleic, linoleic, and linolenic acids, are capable of forming
caprylic acid from their 9-hydroperoxides.[25] Caprylic acid was detected in residual amounts
ranging from 0.07 to 2.13 ppm in some of the fresh palm olein samples.

Principal component analysis (PCA)

Due to the large number of samples (variables) with multiple responses, the use of PCA yields more
robust and reliable results than univariate approaches, especially with respect to the correlations among
analytical parameters.[26] Based on the PCA plot (Figure 1), the first and second principal components
(PCs) had eigenvalues greater than 1 and described 48.39% and 20.70% of the variance, respectively.
Together, the first two PCs represented 69.10% of the total variability. The loading plot of the two PCs
provides information on the correlations among the analytical parameters. The levels of TAG dimers
and the FFAs of the commercial cooking oil samples were found to be negatively correlated with other
measured properties such as smoke points, TPC, DAGs, and oxTAGs along the PC1 axis.
The primary axis was represented by the TPCs, DAGs, smoke point, and FFAs, whereas the
secondary axis was related to the TAG dimers and oxTAGs of the cooking oil samples. The TPCs,

(a) (b)
Figure 1. Principal component (a) score plot and (b) biplot based on quality of commercial cooking oil samples (PPO = pure palm
olein; BPO = blended palm olein; PO = packet oil).
* TPC: total polar compounds; TAG: triacylglycerol; OXTAG: oxidized triacylglycerol monomers; DAG: diacylglycerols; FFA: free fatty
acids
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 1181

oxTAGs, and DAG levels were strongly intercorrelated as these parameters were located in the same
quadrant (right upper quadrant). In this series, the emphasis focused on the quality of different
categories of cooking oils. A biplot is a combination of a score plot and a loading plot. Each of the
dots in the score plot (Figure 1(a)) represents a different type of cooking oil. The biplot (Figure 1(b))
indicated that most of the packet oils contained high TPC, oxTAGs, DAGs, and FFA content. In
addition, most of the pure palm oleins had higher smoke points than the blended oils and packet
oils. In contrast to the packet oils, the pure palm olein samples were found to be better in overall
quality as indicated by the biplot of all analytical parameters.

Conclusion
In this study, the safety and quality of 23 commercial cooking oils from different categories available in
a Malaysian market were successfully evaluated and compared. The presence of the potential markers
for the detection of recycled oil in the commercial oil samples was also determined. Correlations among
analytical parameters were also evaluated through principal component analysis. Most of the packet oils
(pure palm olein) had lower smoke points (< 200°C) than the bottled oils (pure palm olein and blended
palm olein). The total polar compound content of all samples fell within the recommended safety limits
for human consumption (< 25% polar compounds). Unsurprisingly, no TAG oligomers or epoxy, keto,
or hydroxy acids were detected in the fresh oil samples. This result shows that the freshness of oil was
well maintained and that the quality of cooking oils available in Malaysia is satisfactory. This finding
also suggests that the presence of TAG oligomers and epoxy, keto, and hydroxy acids in fresh oil might
serve as a potential indicator of the adulteration of palm olein. The reason is because the molecular
weights of these compounds are high, making them more difficult to remove during the refining
process. The pure palm olein samples were found to be superior to the other types of commercial oil
samples in terms of overall oil quality. The results of this study provide useful insight into the quality of
the commercial cooking oils available in the Malaysian market. This issue is of considerable concern
because cooking oil is widely used during daily meal preparation.

Acknowledgments
The work was supported by the Putra Grant, Universiti Putra Malaysia (Project number UPM/700-1/2/GIPP/2017/
9532400). The authors have declared no conflict of interest.

Funding
This work was supported by the Universiti Putra Malaysia [UPM/700-1/2/GIPP/2017/9532400].

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