Materials Today: Proceedings 42 (2021) 2000–2005

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Effect of different extraction methods on physicochemical properties,

antioxidant activity, of virgin coconut oil
Nameer Khairullah Mohammed ⇑, Ziad Tariq Samir, Mohammed Ahmed Jassim, Sami Khudhur Saeed
Department of Food Science, College of Agriculture, Tikrit University, 34001 Tikrit, Iraq

a r t i c l e i n f o a b s t r a c t

Article history: Since there is a high demand for the virgin coconut oil (VCO) in Iraqi markets due to having consumers
Available online 21 January 2021 recently recognized that VCO has unique nutrition benefits, this study aims to recover the virgin coconut
oil (VCO) using four extraction techniques; fermentation VCO (FVCO), dry VCO (DVCO), enzymatic VCO
Keywords: (EVCO), and the chilling and thawing VCO (CVCO) techniques. The fatty acid composition, physicochem-
Antioxidants ical properties and the antioxidant activities of the VCOs were determined and compared to the commer-
Coconut oil cial refined, bleached and deodorized coconut oil (RBD-CO). Physicochemical properties of extracted
Extraction
VCOs were yield, 54–72%; moisture contents, 0.12–0.16%; refractive index 1.45; viscosity, 48–51 cP; free
Physicochemical properties
Fatty acids
fatty acid, 0.16–0.2 g/100 g; iodine value, 4.17–7,13 g I2/100 g oil; peroxide value 147–259 meq O2/kg oil,
and saponification value 254.10–264.04 mg of KOH/g of oil. Fatty acid composition demonstrated that
lauric acid had the highest content for all the VCOs with the range of 47.95–48.08%. The DPPH radical-
scavenging activity (IC50) for the VCOs were 205.15–248.16 mg/mL and the total phenolic content
(TPC) were37.42–68.12 mg GAE/100 mL.
Ó 2021 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the 3rd International Con-
ference on Materials Engineering & Science.

1. Introduction
Virgin coconut oil (VCO) is usually produced using fresh coco-
nut fruits at room temperature without the use of any solvents
or chemicals [1]. It is derived from the fresh and mature kernels
(12 months after pollination) from the milk of coconut fruits (Cocos
nucifera L.), which belongs to the Arecaceae family (palm) [2]. It
can also be extracted using natural or mechanical techniques with-
out any heat, to prevent any alteration of the oil properties [3]. VCO
was seen to be a colourless oil, with a fresh coconut aroma. It is
mainly used for baking, cooking, preparing infant foods, confec-
tionery and cosmetics. In the cosmetic industry, VCO is popularly
used for most of food applications such as confectionary cooking,
emulsions, bakery and in pharmaceutical and cosmeceutical aims
such as enhancing beauty, promoting hair growth and moisturising
the skin [4]. Generally, coconut oil has been used for industrial and
food-related applications due to be rich in phenolic compounds
which is correlated with the scavenging activity [5] This oil con-
tains a high concentration of lauric acid, which is a medium-
chain fatty acid (MCFA). MCFAs display a good digestibility since
⇑ Corresponding author.
E-mail address: nameer@tu.edu.iq (N.K. Mohammed).
the body uses them as an energy source immediately after their
consumption [6]. They are not stored in the form of body fat [7].
On the other hand, it was seen that the animal fats consisted of
long-chain saturated fatty acids [8]. A majority of the
commercial-grade coconut oils are refined, bleached and deodor-
ized oil (RBD) and produced using different techniques such as
sun drying, smoke drying or their combination. If copra is used
as the primary substrate, the coconut oil that is generated is not
fit for consumption and needs to be purified. Furthermore, an
unsanitary or improper handling or drastic processing of the copra
for extracting and refining oil makes this product susceptible to
oxidative rancidity and aflatoxin contamination [9]. In comparison,
the VCO that is extracted using a wet processing technique from
the coconut milk under a controlled temperature displays better
effects than the copra oil as it retained most of its beneficial prop-
erties. VCO also has many significant biological, nutritional and
medicinal effects [6]. VCO does not undergo any RBD process and
is consumable, in its primary state, without any further chemical
processing. VCO primarily comprises of the medium-chain triglyc-
erides and acts as a potential food item with health benefits. Also,
it is colourless, with no unpleasant taste or rancid odour and does
not consist of any sediment with a fresh, natural coconut scent
[4,10] used different extraction methods for VCO and found that

https://doi.org/10.1016/j.matpr.2020.12.248
2214-7853/Ó 2021 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the 3rd International Conference on Materials Engineering & Science.
Nameer Khairullah Mohammed, Ziad Tariq Samir, Mohammed Ahmed Jassim et al.
by using protease, the VCO had the highest extraction yield and the
greatest amount of unsaturated fatty acids, while the fermentation
method had the highest contents a-tocopherol and phenolic com-
pounds. In addition, the study of [11] showed that VCO obtained by
solvents had the highest oil yield while the oil extracted by fer-
mentation was the lowest and the oil extracted by freezing and
thawing was the most preferred in sensory attributes. However,
from the previous studies it is clear that extraction method have
significant impacts on the quality of VCO. Because of its beneficial
therapeutic value, VCO gained a lot of popularity in the past few
years. Day-by-day consumers are becoming more aware and have
started demanding edible oils which are natural and not chemi-
cally processed. Hence, studies that investigated the chemical com-
position are of a lot of interest to the consumers who demand
information related to the properties and characteristics of the
types of VCOs that are available. To the best of our knowledge,
there is no report yet on the extraction of VCO in Iraqi industries.
Therefore, it is important to investigate the VCO properties that
obtained in this country. Very few researchers have determined
the chemical properties of the VCOs and RBD-CO which are avail-
able in the Iraqi markets. Hence, the objective of this study was
to extract the VCO by four different extraction methods namely
fermentation, dry, enzymatic, and the chilling and thawing meth-
ods and analysed their quality compared to commercial coconut
oil. For this purpose, the total flavonoid content, total phenolic con-
tent, and the antioxidant properties of the VCOs using biochemical
and chemical assays were measured as well as physicochemical
properties including yield, refractive index, viscosity, moisture
contents, free fatty acid, saponification value, iodine value, perox-
ide value and fatty acid composition in VCO extracted in Iraq
2. Material and methods
The researchers selected fully matured coconuts (10–11-
month-old) from a local grocery in Tikrit, Iraq, for extracting the
VCOs. The refined, bleached and deodorized coconut oil (RBDCO)
was purchased from (Carrefour) the local grocery in Erbil, Iraq for
comparison. Chemicals that were used in the study were pur-
chased from Sigma Aldrich (Germany) and were of analytical
grade.
2.1. Preparation of coconut milk
Before extracting the coconut milk, good coconut fruits were
selected, dehusked, deshelled and the testa was removed, washed,
grated and finally, the coconut milk was extracted. Using a special
tool, the shell of the nut was removed, and the kernels were
scooped out using the knife. The testa of the coconut kernel was
also removed using a testa remover machine. The kernel that
was freed from the testa was fed into a mechanical grating
machine (which contained rotating blades). The grated coconut
meat was used for extracting the coconut milk with the help of
the manually-operated hydraulic press. The milk that was
extracted after extraction 1 was stored and the residue was further
used for extractions 2 and 3. Finally, all the milk extracts were
pooled and stirred vigorously for some time.
2.2. Extraction of VCO
VCO is extracted from coconut milk or coconut meat. Extraction
of oil can be done by four different techniques namely, fermenta-
tion method (FVCO), enzymatic method (EVCO), dry method
(DVCO), chilling and thawing method (CVCO) for producing VCOs.
The VCOs were extracted using the procedure described by Mansor
et al. [12] with a few modifications.
2001
Materials Today: Proceedings 42 (2021) 2000–2005
2.2.1. Fermentation method
The fresh coconut milk, extracted above, was diluted using
equal parts of distilled water. 2 g of Baker’s yeast (Saccharomyces
cerevisiae) was added to 1 L of this mixture as the inoculum for fer-
mentation. The mixture was homogenised by mixing thoroughly.
This mixture was left to stand at room temperature for 36 h, which
helped in separating the water and oil layers. The upper oil layer
was decanted. This process was repeated 3 times and the coconut
oil was refrigerated for further use.
2.2.2. Chilling and thawing method
In this study, the researchers used the chilling and thawing
technique as described by [13] with a few modifications. The coco-
nut milk that was extracted using the above-mentioned step was
centrifuged for 10 min at 3600g. The upper cream layer was
removed after chilling for 24 h at 5 °C. The chilled cream slowly
thawed in a water bath at 50 °C for extracting the coconut oil. This
process was repeated thrice and the extracted oil was refrigerated
for further use.
2.2.3. Enzymatic treatment
The coconut oil was extracted using the enzymatic technique
described by [13] with a few modifications. Papain enzyme (0.1%,
w/w) was added to the extracted coconut milk, and the mixture
was homogenised by stirring. This solution was allowed to stand
at 55 °C for 3 h, as this was the optimal temperature for papain
activity. Thereafter, this mixture was centrifuged for 25 min at
4900g for extracting the coconut oil. This process was repeated
thrice and the oil was refrigerated for further use.
2.2.4. Fresh-dry process
In this technique, the white coconut meat was shredded and
then dried in the aerated oven (35 °C) for 48 h. During the drying
process, the sample was stirred to ensure homogenous drying
and for preventing the scorching of the meat. Thereafter, the sam-
ple was screw pressed for extracting the oil. The oil that was col-
lected was filtered through the Whatman No. 1 filter paper to
remove any debris. This process was repeated thrice and the oil
was refrigerated for further use.
2.3. Physical analysis
The colour of the VCO samples were determined by Hunter Lab
Colorimeter (Hunter Associates Laboratory Inc., Virginia, USA). The
amount of moisture in the samples was determined using the
method of AOAC [14]. The Refractive Index (RI) of the VCO samples
were assessed by Abbé refractometer (Bellingham & Stanley, U.K.).
The viscosity was measured using the AR G2 Rheometer (TA Instru-
ments, US).
2.4. Chemical analysis
The Free Fatty Acid (FFA) values were determined using the
IUPAC [15] technique. The Iodine Value (IV) of the samples was
determined using the Wij method (AOCS) [16]. The researchers
determined the Saponification Value (SV) of the oil samples using
the IUPAC technique [15].
2.5. Fatty acid methyl esters (FAME)
The researchers prepared the FAMEs of the oil samples using
the method described by Hartman and Lago [17] with a few mod-
ifications for ensuring the samples were volatile so that they could
be analysed using the GC-FID instrument.
Nameer Khairullah Mohammed, Ziad Tariq Samir, Mohammed Ahmed Jassim et al.
2.6. Antioxidant activity
The total phenolic compounds of the VCO sample were by the
Folin–Ciocalteu Reagent (FCR), based on the method described by
Singleton [18] with a few modifications. The radical scavenging
activities of the 5 VCO and RBD-CO samples as the described
method of Blois [19] using the DPPH assay.
2.7. Statistical analysis
Probability of p > 0.05 was regarded as significant. The statisti-
cal analyses were carried out with the help of the MINITAB 16 sta-
tistical software (Minitab Inc., State College, PA, USA).
3. Results and discussion
3.1. Physicochemical properties
The effects of extraction methods including FVC, EVC, CVC and
DVC on the VCO yields were investigated. All the VCO yield had
notably significant differences (p < 0.05). The highest yield was
detected for the FVCO (72%), followed by CVCO (70%), EVCO
(62%) and DVCO oil (54%) (Table 1). The low oil yield from the
fresh-dry method may be attributed to the limitation of the screw
pressed pressure on the coconut meat. The enzymatic treatment
showed the highest yield among all the extraction methods. This
result was agreed with [7,13] who reported the application of
enzymatic treatment for the VCO production and obtained the
highest yield compared to other techniques. Other study by [8]
was also reported extraction VCO by aid of crude protease extract
from white shrimp resulted in higher yield up to 93%. It was noted
that the VCO yield depended on the extraction techniques along
with many additional factors like the age of the coconuts and
copra, time of harvesting and the location of the plantation [4].
One of the major factors which affected the quality of the VCO pro-
duct was moisture. The moisture content of the VCOs that were
extracted from the coconut milk using different techniques was
compared to the RBD-VCO sample, and results are presented in
Table 1. The moisture content of the VCOs should be lower than
0.1% according to the APCC standards (APCC 2009). VCO samples
extracted using the fermentation process and enzymatic treatment
showed similar moisture content (0.15%). The DVCO was found to
be the lowest moisture content among all the VCO samples with
significant differences (p < 0.05). This may be due to the drying
conditions that significantly remove the water components from
the VCO. As shown in the table, the RBD-VCO sample exhibited
the highest moisture content (0.17%, p < 0.05) amongst all the sam-
ples. Higher moisture content reduces the shelf-life of the VCO
samples. It also increases the hydrolysis and oxidation levels,
which increases the free fatty acid content and the hydrolytic ran-
cidity of the samples [21]. The viscosities of VCO samples extracted
using four different extraction methods were given in Table 1. The
Table 1
Chemical properties of VCO extracted by four different extraction methods, commercial coconut oil and Codex standard.
Physiochemical properties FVCO EVCO
FFA (as oleic %) 0.2 ± 2.00e 0.2 ± 1.01c
IV (g of I2/100 g of oil) 6.05 ± 0.03c 4.33 ± 0.03b
PV (meq O2/kg oil) 2.59 ± 0.00b 2.34 ± 0.00c
SN (mg of KOH/g of oil) 262.55 ± 0.07c 259.55 ± 0.12d
Yield% 72 ± 2.00d 62 ± 1.04b
Moisture content 0.15 ± 2.00c 0.15 ± .50c
Viscosity (cP) 51 ± 0.03d 49 ± 0.05b
RI 1.45 ± 0.04a 1.45 ± 0.05a
Means ± SD (n = 3). Different lowercase superscripts in the same column indicate significant differences (P < 0.05).
2002
Materials Today: Proceedings 42 (2021) 2000–2005
lowest viscosity was recorded for the VCO obtained by dry method
and RBD-CO while the highest value was for the fermentation
treatment with significant differences (p < 0.05). The range of the
viscosity records was similar to that reported by [12] VCO
extracted with dry method, fermentation method and enzymatic
method. These techniques are applied without any effect on the
compositions of the oils. It is well known that VCO has medium-
chain acid which might be the reason behind its lower viscosity
compared to the oils consist of long-chain fatty acid [21]. The RI
of VCO samples resulted in negligible differences values ranging
between a minimum values of 1.332 to the maximal value of
1.345 with no significant differences (p < 0.05) Table 1. These RI
values ranged between the standard values defined by APCC [22].
Similar results were observed by Ghani et al. [4] who observed that
the VCOs extracted from the coconut milk as the substrate using
different techniques did not show any significant difference in
their RI values. The RI value was related to the degree of unsatura-
tion present in the lipid sample. It increased proportionally with an
increasing number of double bonds in the sample. The similar RI
values confirmed that VCO has a similar saturation degree due to
the double bond number. The RI factor also helped in determining
the purity of the oil sample as every substance exhibits a particular
and different RI value [21]. The colour of the VCO samples was an
important parameter which could affect the acceptability of the
final sample as a food product by the consumers. The researchers
compared the various associated factors like redness (a*), yellow-
ness (b*), and brightness (L*) of the VCO samples (Table 3). The
brightness and yellowness of the samples need to be especially
determined. The results indicated that the L* values of the VCO
samples were similar without any significant difference
(p < 0.05), as the VCO must be transparent and almost colourless.
The oil colour can be affected by numerous parameters like the
variety and type of the coconut and even its ripeness. Table 3
shows the L* had slight differences while the highest brightness
was achieved by (EVCO) (99.725 ± 0.219), followed by (CVCO)
(99.34 ± 0.28), (FVCO) (99.335 ± 0.12), (RBD-VCO) (98.91 ± 0.15),
and (DVCO) (98.855 ± 0.02). These findings were in accordance
with that reported by [12] VCO samples extracted by different
techniques showed similar colour properties. Similar results were
given by [23] VCO extracted with enzymatic treatment showed
the highest brightness compared to other methods. On the other
hand, the lowest yellowness value was noted for the EVCO sample
(0.735 ± 0.021), while the DVCO sample showed the highest value
(2.755 ± 0.120), followed by the FVCO (2.105 ± 0.03), CVCO
(1.78 ± 0.02) and the RBD-VCO (1.5 ± 0.05) oil samples. The VCO
that was extracted using the mechanical press showed a lighter
colour and a low FFA content. The refining process of the vegetable
oils that were extracted using the mechanical press or solvent
extraction techniques included the removal of coloured particu-
lates by bleaching. The oils extracted using the Supercritical Fluid
(SFE) extraction method were light in colour. Different colours
were attributed to an increasing amount of the dissolved colorants
CVCO DVCO RBD-CO CODEX
0.19 ± 1.20d 0.16 ± 1.03a 0.17 ± 0.04b Max 0.2
7.13 ± 0.01d 4.17 ± 0.07a 5.70 ± 0.05c 4.1–11
2.19 ± 0.00e 1.467 ± 0.00a 2.18 ± 0.00d Max 3
254.10 ± 0.01e 264.04 ± 0.02b 264.38 ± 0.06a 250 – 260
70 ± 0.80c 54 ± 0.05a –
0.14 ± 1.30b 0.12 ± 1.02a
50 ± 2.03c 48 ± 1.04a 48 ± 1.30a –
1.45 ± 0.04a 1.45 ± 0.05a 1.45 ± 0.02a –
Nameer Khairullah Mohammed, Ziad Tariq Samir, Mohammed Ahmed Jassim et al. Materials Today: Proceedings 42 (2021) 2000–2005

in the extracted oil samples. Table 2 presents the values for the Table 3
Free Fatty Acid (FFA) content of the oil samples using different Color (Hunter Lab L* a* b*) value VCO for five samples of VCOs and RBD-VCO.

techniques. These FFA values ranged between 0.16 and 0.2 (lauric
acid) (Table 2). All the VCO samples in the present study had FFA
aligned with the Codex standards [24] good coconut oil must have
FFA content less than 0.2. The highest FFA level was noted in the
FVCO and EVCO samples, whereas the lowest FFA content was seen
in the DVCO samples. These samples showed a significant differ-
ence in their lauric acid content (p < 0.05). Similar findings were
reported by [25] FFA was higher for VCO obtained by enzymatic
method compared to other methods. Also results were reported
by [8] also found that fermentation extraction method resulted
in VCO with high FFA. High FFA content was seen when they used
a fermentation process for VCO extraction. Furthermore, the VCO
samples that were extracted using the enzymatic hydrolysis of
the esterified lipids showed high FFA content. The phospholipases
and lipases in the coconut milk were still active, which increased
the lipid hydrolysis rate [8]. Marina, [26] noted similar results as
they did not note any changes in the distribution of the fatty acids
in the VCO and RBD-VCO samples. The variation in the FFA content
of the VCO oils could be attributed to the different processing tech-
niques that were used. According to [23] the coconut oils with a
higher moisture content showed a higher FFA value. All vegetable
oils naturally show low FFA content.
Furthermore, during extraction or storage, the FFA content
could increase because of the reaction between the oil and residual
water. This was the result of hydrolytic rancidity, which hydrol-
ysed the ester, either chemically or enzymatically (using lipases).
A high FFA content was responsible for the development of unde-
sirable aromas and flavour in the oil samples. IV was used for mea-
suring the degree of unsaturation present in the oils and fat
samples. It was expressed as the grams of iodine that was absorbed
by 100 g fats under the conditions used for the test. In this study,
the IV of the VCO samples ranged between 4.17 and 7.13 g
I2/100 g oil. This low IV content showed that the VCO has a higher
saturation level, which increased its resistance to the oxidative
rancidity [27]. All the VCO samples showed a significant difference
between the IV values (p < 0.05). The CVCO sample showed the
highest IV whereas the DVCO sample showed the lowest IV
(Table 1). The IV values displayed by the samples ranged between
the limits defined by the Codex [24] and APCC [22] standards for
the edible coconut oils. The IV value of the oil obtained by the
enzymatic method was in accordance with the study described
by [20]. The EVCO sample showed an IV of 4.89 g I2/100 g oil.
Peroxide Value (PV) used for determining the concentration of
hydroperoxides and peroxides formed during the initial lipid oxi-
dation stage. The peroxides in the oil samples reflected the oxida-
tive level and its likelihood of becoming rancid. Table 1 presents
the PV of the VCO samples. A significant difference was noted
amongst the oil samples (p < 0.05). The highest PV value was seen
in the FVCO (2.59 meq. O2/kg oil), whereas the DVCO sample
showed the lowest PV of 1.59 meq. O2/kg oil (Table 1). These values
Color L* a* b*
Oil samples
DVCO 98.855 ± 0.24c 1.245 ± 0.01d 2.755 ± 0.04a
FVCO 99.335 ± 0.07b 0.94 ± 0.00a 2.105 ± 0.05b
EVCO 99.725 ± 0.00a 0.305 ± 0.00b 0.735 ± 0.01d
CVCO 99.34 ± 0.05b 1.14 ± 0.18d 1.78 ± 0.03c
RBD-VCO 98.91 ± 0.03c 0.895 ± 0.00c 1.5 ± 0.01e
Means of triplicate measurements in the same column ± standard deviation with
different letters are significantly different (p < 0.05).
were within the APCC standards [22] (3.0 meq. O2/kg). The PV
values for all the oils were relatively low, which indicated that
the samples were highly resistant to oxidation. However, the
Codex standard [24] for the maximal PV value of VCO samples
was 15 meq. O2/kg oil. Hence, the PV of the oils indicated that all
samples were fresh. In theory, the VCOs must exhibit a low oxida-
tion rate because of their lower unsaturated fatty acid content [28].
The unsaturated fatty acids react with the oxygen molecules to
form peroxides. The oils having a higher PV were unstable and
easily turned rancid. As stated by [29] fats or oils contained higher
degree of unsaturated fatty acids are more vulnerable to the
oxidation.
Table 1 presents the Saponification Values (SV) of the VCOs. The
SV was a measure of the mean molecular weights of the fatty acids
present in the oils. The SV for all the VCOs investigated in this
study ranged between 254.10 and 264.38 mg KOH/g (Table 1).
The Codex standard [24] stated that the SV for the edible coconut
oils must range between 248 and 265 mg KOH/g oil. The EVCO
sample showed the maximal SV of 264.38 mg KOH/g. These results
were similar to those observed by [20] who noted that the SV of
the VCOs extracted from the coconut milk using enzymatic tech-
niques was 262.72 mg KOH/g oil. Higher the SV of the oils, shorter
were the fatty acids present in their glycerol backbone. In compar-
ison to the other vegetable oils, the VCOs showed a higher SV,
which indicated that the VCO contained a higher number of
short-chain fatty acids.
3.2. Fatty acid methyl esters (FAME) compositional analysis
The fatty acid analysis provided information regarding the fatty
acid distribution in the oil and fat samples. Table 2 presents the
fatty acid composition of the RBD-VCO and 5 VCO samples that
were extracted using different techniques. The results indicated
that the fatty acid composition of the VCOs was in accordance with
the APCC and Codex standards for coconut oil. As shown in Table 2,
the concentration of lauric acid (C12:0) in the VCO samples ranged
between 47.95% and 48.83%, which was as per the standard values.
The total MCFA concentration in the VCOs (C8, caprylic acid; C10,
capric acid and C12, lauric acid) was 61.77%, 62.13%, 61.75%,

Table 2
Fatty acid composition of VCO extracted by four different types of extraction methods, commercial coconut oil, Codex standard and APCC standard.

Fatty Acid (%) FVCO EVCO CVCO DVCO RBD-VCO APCC Standard (%) CODEX Standard (%)
C8:0 7.64 ± 0.01c 7.43 ± 0.01d 7.5 ± 0.02c 7.97 ± 0.01b 7.88 ± 0.01a 4–10 4.6–10.0
C10:0 6.18 ± 0.01b 6.02 ± 0.01d 6.01 ± 0.01c 6.31 ± 0.00a 6.35 ± 0.00a 4–8 5.0–8.0
C12:0 47.95 ± 0.01d 48.68 ± 0.01e 48.24 ± 0.01a 48.83 ± 0.01c 48.74 ± 0.03b 45–56 45.1 53.2
C14:0 18.58 ± 0.01d 18.94 ± 0.01a 18.85 ± 0.01b 18.7 ± 0.01c 18.52 ± 0.01e 16–21 16.8–21.0
C16:0 9.04 ± 0.00b 9.08 ± 0.01a 8.99 ± 0.00b 8.59 ± 0.02d 8.68 ± 0.01c 7.5–10.2 7.5–10.2
C18:0 3.16 ± 0.20a 3.13 ± 0.00a 3.1 ± 0.00a 3.27 ± 0.01a 3.09 ± 0.01a 2–4 2.0–4.0
C18:1 6.07 ± 0.00b 6.17 ± 0.00a 6.08 ± 0.01d 5.36 ± 0.01c 5.62 ± 0.01c 4.5–10 5.0–10.0
C18:2 1.38 ± 0.00a 1.36 ± 0.00b 1.23 ± 0.00c 0.97 ± 0.00d 1.12 ± 0.01e 0.7–2.5 1.0–2.5

Means within each column with different superscript are significantly different at (p < 0.05).

2003
Nameer Khairullah Mohammed, Ziad Tariq Samir, Mohammed Ahmed Jassim et al.
63.11% and 62.97% for the FVCO, EVCO, CVCO, DVCO and RBD-VCO,
respectively. The highest lauric acid was obtained from the sample
DVCO with 48.83% ± 0.01% with significant differences between all
the VCO samples (p < 0.05). These results were similar to the fatty
acid composition of the VCOs extracted using the integrated wet
technique (62.6%–63.7%) that was reported earlier [30]. On the
other hand, the FVCO, EVCO, CVCO, DVCO and the RBD-VCO sam-
ples showed a long-chain fatty acid (C14-C18) content of 30.78%,
31.15%, 30.94%, 30.56% and 30.29%, respectively. The unsaturated
linoleic acid (C18:1) concentration for the VCO samples ranged
between 5.36% and 6.17%, while the linoleic acid (C18:2) ranged
from 0.97% to 1.12%. The coconut oil that was extracted using dif-
ferent techniques in this study showed a different fatty acid com-
position. This was attributed to the oxidation of the sample which
occurred due to the hydraulic process, type of extraction method
and the nature or structure of fatty acids. The results indicated that
the various extraction methods could affect the fatty acid compo-
sition of the oil samples.
3.3. Antioxidant activity
Total phenolics content (TPC) are secondary metabolites widely
distributed in plants. TPC for the VCO extracted by different pro-
cessing method, fermentation method, enzymatic, chilling and
drying technique and refined, bleached, deodorised oil (RBD) were
evaluated. The TPCs have been detected for the VCO samples in the
range of (37.42 to 68.12 mg GAE/100 mL) and presented in Fig. 1.
The CVCO sample showed the highest TPC (68.12 mg GAE/100 mL
oil), whereas the RBD-VCO showed the lowest TPC (37.42 mg
GAE/100 mL). This finding can be due to the low temperature
applied for the CVCO extraction with chilling method resulted in
high phenolic compounds, compared to the DVCO obtained with
dry method extraction. Furthermore, the FVCO sample showed a
high TPC (61.98 mg GAE/100 mL oil), which was similar to the
results noted by [31] who noted that the VCO extracted using
the fermentation technique showed a high TPC (59.44 mg
GAE/100 mL oil). Seneviratne and Dissanayake [32] observed that
the VCOs extracted using different techniques showed a TPC rang-
ing between 91 and 618 mg GAE/100 mL. It has been reported that
the phenolic compounds were thermally unstable if high tempera-
tures are used for extraction of coconut oil. The low total phenolic
Fig. 1. Total phenolic content (TPC), total flavonoid compounds (TFC) and antioxidant activity by (DPPH) in four types of VCO and commercial coconut oil.
2004
Materials Today: Proceedings 42 (2021) 2000–2005
content observed in RBD-CO maight be due to the multi steps
applied for refining the oil and also due to the exposure of oil to
high heat during the extraction process [33]. The health benefits
of the VCOs were attributed to their potent phenolic compounds
that could be conserved using the traditional wet production tech-
nique [34]. The researchers also determined the antioxidant capac-
ity of the VCO samples. They used the DPPH technique with the
stable, organic 1,1-diphenyl-2-picrylhydrazyl radical for determin-
ing the free radical scavenging activity (expressed as IC50). This was
described as the number of antioxidants required for decreasing
the initial DPPH concentration by 50%. Thus, lower the IC50 value
of the VCO, higher was its antioxidant activity. This assay is based
on the colorimetric change that DPPH undergoes in the presence of
an antioxidant. It can be quantified by measuring its absorbance,
wherein a decrease correlates with DPPH scavenging activity of
the antioxidant. The IC50 for the VCO samples were in the range
of (205.15–248.16) mg/mL Fig. 1. The IC50 value for the FVCO
was demonstrated the highest radical activity 205.15 mg/mL while
the lowest radical activity was recorded for the DVCO 248.16 mg/
mL with significant difference (p < 0.05). Similar results were
reported by [25] who found that VCO obtained by fermentation
method had highest antioxidant activity among different extrac-
tion methods. This finding also was in alignment to the study
reported by [2] VCO extracted by fermentation treatment had
highest radical scavenging activity followed by the chilling method
compared to the RPD oil with the lowest activity. The previous
study carried out by Librado and Von Luigi (2013) using DPPH
assay has reported lower free radical scavenging activity of VCO
(IC50) is at 313.46 mg/mL. The processing conditions used for the
VCO samples contributed to their free radical scavenging and
antioxidant activities.
4. Conclusions
This study was conducted for determining the manner in which
the analysis, composition and the properties of the VCO samples
could increase the industrial application of these samples. The
researchers evaluated the effect of the extraction technique on
the quality of the oil that was extracted. In this study, they deter-
mined the properties of the VCOs that were extracted using 4 tech-
niques, i.e., the enzymatic, fermentation, drying and chilling-and-
Nameer Khairullah Mohammed, Ziad Tariq Samir, Mohammed Ahmed Jassim et al.
thawing techniques. The results of the physicochemical properties
of VCOs including the RBD-VCO were compared to the Codex and
APCC standard for edible coconut oil and revealed values within
the limit The results indicated that the antioxidant activity and
the TPC of the VCOs were statistically significant (p  0.05). From
an economic point of view, the highest yield was recorded for
the FVCO, which was considered as a vital factor that affected
the cost of the oil products. On the other hands, the quality of
the oil in terms of its health benefits and biological activities CVCO
showed better quality among the other oils.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
References
[1] A. Rohman et al., Virgin coconut oil: extraction, physicochemical properties,
biological activities and its authentication analysis, Food Rev. Int. (2019) 1–21.
[2] A.M. Marina, Y.B. Che Man, S.A.H. Nazimah, I. Amin, Chemical properties of
virgin coconut oil, J. Am. Oil Chem. Soc. 86 (4) (2009) 301–307.
[3] P. Soo et al., Enzymatic and mechanical extraction of virgin coconut oil, Eur. J.
Lipid Sci. Technol. 122 (5) (2020) 1900220.
[4] N.A.A. Ghani, A.-A. Channip, P. Chok Hwee Hwa, F. Ja’afar, H.M. Yasin, A.
Usman, Physicochemical properties, antioxidant capacities, and metal contents
of virgin coconut oil produced by wet and dry processes, Food Sci. Nutr. (2018).
[5] S.N.A. Abd Rashid, M. Misson, H. Yaakob, N.A. Latiff, M.R. Sarmidi, Addition of
virgin coconut oil: influence on the nutritional value and consumer acceptance
of dark chocolate, Trans. Sci. Technol. 4 (3–3) (2017) 426–431.
[6] M. DebMandal, S. Mandal, Coconut (Cocos nucifera L.: Arecaceae): in health
promotion and disease prevention, Asian Pacific J. Tropical Med. 4 (3) (2011)
241–247.
[7] R. Handayani, J. Sulistyo, R.D. Rahayu, Extraction of coconut oil (Cocos nucifera
L.) through fermentation system, Biodiversitas J. Biol. Divers. 10 (3) (2009).
[8] T. Senphan, S. Benjakul, Chemical compositions and properties of virgin
coconut oil extracted using protease from hepatopancreas of Pacific white
shrimp: virgin coconut oil extracted using shrimp protease, Eur. J. Lipid Sci.
Technol. 118 (5) (2016) 761–769.
[9] P. Nakpong, S. Wootthikanokkhan, High free fatty acid coconut oil as a
potential feedstock for biodiesel production in Thailand, Renew. Energy 35 (8)
(2010) 1682–1687.
[10] R. Prapun, N. Cheetangdee, and S. Udomrati, ‘‘Characterization of virgin
coconut oil (VCO) recovered by different techniques and fruit maturities.,” Int.
Food Res. J., 23(5), 2016.
[11] J. Ndife, D. Obot, and K. Abasiekong, ‘‘Quality Evaluation of Coconut (Cocos
nucifera L) Oils Produced by Different Extraction Methods,” Asian Food Sci. J.,
pp. 1–10, 2019.
[12] T.S.T. Mansor, Y.B.C. Man, M. Shuhaimi, M.J.A. Afiq, F.K.M.K. Nurul,
Physicochemical properties of virgin coconut oil extracted from different
processing methods, Int. Food Res. J. 19 (3) (2012) 837.
2005
Materials Today: Proceedings 42 (2021) 2000–2005
[13] U. Patil, S. Benjakul, Coconut milk and coconut oil: their manufacture
associated with protein functionality, J. Food Sci. 83 (8) (2018) 2019–2027.
[14] AOAC, ‘‘Official Methods of Analyzes,” in Association of Official Analytical
Chemist, 1995.
[15] IUPAC, ‘‘Standard Methods For The Analysis Of Oils,” in Fats And Derivatives,
1979.
[16] AOCS, ‘‘Method Cd 3-25. Estimation of saponification value.” Official Methods
and Recommended Practices of the American Oil Chemist’s Society,
Champaign IL, USA, 2004.
[17] L. Hartman, Rapid preparation of fatty acid methyl esters from lipids, Lab.
Pract. 22 (1973) 475–476.
[18] V.L. Singleton, R. Orthofer, R.M. Lamuela-Raventós, [14] Analysis of total
phenols and other oxidation substrates and antioxidants by means of folin-
ciocalteu reagent, in: Methods in enzymology, Elsevier, 1999, pp. 152–178.
[19] M.S. Blois, Antioxidant determinations by the use of a stable free radical,
Nature 181 (4617) (1958) 1199.
[20] T. Senphan, S. Benjakul, Comparative study on virgin coconut oil extraction
using protease from hepatopancreas of pacific white shrimp and alcalase, J.
Food Process. Preserv. 41 (1) (2017) e12771.
[21] U. Patil and S. Benjakul, ‘‘Use of Protease from Seabass Pyloric Caeca in
Combination with Repeated Freeze–Thawing Cycles Increases the Production
Efficiency of Virgin Coconut Oil,” Eur. J. Lipid Sci. Technol., p. 1800460, 2019.
[22] A. and P. C. C. APCC, ‘‘APCC standards for virgin coconut oil,” 2009.
[23] Y.B. Che Man, M.I.B. Abdul Karim, C.T. Teng, Extraction of coconut oil with
Lactobacillus plantarum 1041 IAM, J. Am. Oil Chem. Soc. 74 (9) (1997) 1115–
1119.
[24] A.C. Codex, Codex, alimentarius commission, Codex Standard for Named
Vegetable Oils, Food and Agriculture Organization of the United Nations,
World Health Organization (WHO), 2001.
[25] N. T. Oseni, W. Fernando, R. Coorey, I. Gold, and V. Jayasena, ‘‘Effect of
extraction techniques on the quality of coconut oil,” African J. Food Sci., 11(3),
pp. 58–66, 2017.
[26] A.M. Marina, Y.B. Che Man, I. Amin, Virgin coconut oil: emerging functional
food oil, Trends Food Sci. Technol. 20 (10) (2009) 481–487.
[27] A. Rohman, Y. B. Che Man, A. Ismail, and P. Hashim, ‘‘Monitoring the oxidative
stability of virgin coconut oil during oven test using chemical indexes and FTIR
spectroscopy.,” Int. Food Res. J., 18(1), 2011.
[28] D. Moigradean, M.-A. Poiana, I. Gogoasa, Quality characteristics and oxidative
stability of coconut oil during storage, J. Agroaliment. Process. Technol. 18 (4)
(2012) 272–276.
[29] S. Cunha, M. Oliveira, Discrimination of vegetable oils by triacylglycerols
evaluation of profile using HPLC/ELSD, Food Chem. 95 (3) (2006) 518–524.
[30] M.A. Hamid, M.R. Sarmidi, T.H. Mokhtar, W.R.W. Sulaiman, R.A. Aziz,
Innovative integrated wet process for virgin coconut oil production, J. Appl.
Sci. 11 (13) (2011) 2467–2469.
[31] C. Ngampeerapong, V. Chavasit, R.W. Durst, Bioactive and nutritional
compounds in virgin coconut oils, Malays. J. Nutr. 24 (2) (2018) 257–267.
[32] K. N. Seneviratne and D. M. Sudarshana Dissanayake, ‘‘Variation of phenolic
content in coconut oil extracted by two conventional methods,” Int. J. food Sci.
Technol., 43(4), pp. 597–602, 2008.
[33] A.M. Marina, Y.B. Che man, S.A.H. Nazimah, I. Amin, Antioxidant capacity and
phenolic acids of virgin coconut oil, Int. J. Food Sci. Nutr. 60 (sup2) (2009) 114–
123.
[34] S.P. Illam, A. Narayanankutty, A.C. Raghavamenon, Polyphenols of virgin
coconut oil prevent pro-oxidant mediated cell death, Toxicol. Mech. Methods
27 (6) (2017) 442–450.