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Extraction of Aril Oil from Gac (Momordica cochinchinensis Spreng) Using

Supercritical Carbon Dioxide

Article in Journal of Food Processing and Preservation · September 2016

DOI: 10.1111/jfpp.13122

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 Journal of Food Processing and Preservation ISSN 1745-4549

EXTRACTION OF ARIL OIL FROM GAC (MOMORDICA

COCHINCHINENSIS SPRENG) USING SUPERCRITICAL
CARBON DIOXIDE
SUTHIDA AKKARACHANEEYAKORN1,3, APINYA BOONRATTANAKOM1, PORNCHANOK PUKPIN1,
SAMAPORN RATTANAWARAHA1 and NAKARIN MATTAWEEWONG2
1
Department of Agro-Industrial, Food, and Environmental Technology, Faculty of Applied Science, King Mongkut’s University of Technology North
Bangkok, 1518 Pracharat I Road, Wongsawang, Bangsue, Bangkok 10800, Thailand
2
Division of Herb and Thai Tradition Medicine, Department for Development of Thai Tradition and Alternative Medicine, Ministry of Public Health, 117
Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand

3
Corresponding author. ABSTRACT
TEL: 166 2555 2000 ext. 4722;
FAX: 166 2587 8257; The objectives of this research were to determine the suitable conditions and
EMAIL: suthida.a@sci.kmutnb.ac.th mathematical equations to predict the efficiency of aril oil extraction from gac
(Momordica cochinchinensis Spreng), including the amounts of b-carotene, lyco-
Received for Publication October 2, 2015
pene, iodine value, acidity, and peroxide in the oil, using supercritical carbon
Accepted for Publication May 27, 2016
dioxide. The experiments were planned using central composite design (CCD) for
doi:10.1111/jfpp.13122 two factors, temperature (31, 35, 45, 55, and 60C) and pressure (130, 150, 200,
250, and 271 bar), and the optimal conditions for oil extraction were 50–60C and
200–250 bar, which resulted in 80–100% extraction efficiency, 30–50 mg
b-carotene per 100 g, 10–20 mg lycopene per 100 g of oil, 60–80% iodine values,
and an acidity of 0–4 mg potassium hydroxide. In comparing the oil extraction
efficiency and the amounts of b-carotene, lycopene, iodine value, and acidity,
there was a discrepancy (%D) of <10% between the experimental values and the
values predicted from a mathematical model at 55C and 220 bar, indicating that
the mathematical model was able to predict the extraction efficiency and the
amounts of b-carotene, lycopene, iodine value, and acidity very well. Moreover,
when the pressure was increased, the extraction efficiency also increased, and
when the temperature was increased at a pressure of 150–200 bar, the extraction
efficiency decreased. These increases in temperature and pressure resulted in
greater amounts of b-carotene and lycopene in the aril oil from Momordica
cochinchinensis Spreng.

PRACTICAL APPLICATIONS
The extraction of aril oil from Momordica cochinchinensis Spreng using supercriti-
cal fluid was studied. The optimal conditions for the extraction of aril oil from
Momordica cochinchinensis Spreng using a supercritical fluid were 52–55C and
200–250 bar, which resulted in an extraction efficiency of 80–100%, 30–50 mg b-
carotene per 100 g, 10–20 mg lycopene per 100 g, 60–80% iodine values, and an
acidity of 0–4 mg potassium hydroxide per gram. Supercritical fluid extraction
resulted in a better extraction efficiency of oil with higher amounts and better
chemical properties of carotenoids compared to using a screw press.

INTRODUCTION
Gac (Momordica cochinchinensis Spreng) is a member of the
Cucurbitaceae family, which includes bitter gourd and
cucumber. It is a wild herbaceous climber found in mixed
deciduous and evergreen forests throughout Southeast Asia
and is also found in China, Bangladesh, and some parts of
Australia (Sone et al. 2010; Kubola and Siriamornpun
2011). The gac fruit is ovoid in shape, densely covered with

Journal of Food Processing and Preservation 2017; 41: e13122; V

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EXTRACTION OF ARIL OIL FROM GAC
3- to 4-mm long spiny tubercles and turns orange or red
once ripe. It can weigh from 500 g up to 2 kg, and the black
seeds inside are covered by red flesh that contains high-
value phytochemicals such as lycopene and b-carotene,
which are antioxidants. Fresh gac aril contains lycopene and
b-carotene at concentrations of 400–1,500 and 100–2,000
mg g21, respectively (Aoki et al. 2002; Betty et al. 2004;
Vuong et al. 2005). Research has shown that lycopene can
help reduce the risk of cancer, heart disease, and diabetes
(Goula and Adamopoulos 2005; Sone et al. 2010; Kubola
and Siriamornpun 2011; Martins, et al. 2015). The body
converts b-carotene into vitamin A, which is essential for
health. Aril oil from gac fruit (Momordica cochinchinensis
Spreng) is nutritious and contains lycopene and b-carotene.
In addition, the gac oil of Momordica cochinchinensis Spreng
aril contains fatty acids that are beneficial to the body, such
as oleic acid, palmitic acid, and linoleic acid (Chuyen et al.
2015).
There are several methods used to extract oil from the aril
of Momordica cochinchinensis Spreng. Screw press and
hydraulic oil extractions use force to press or squeeze,
thereby creating compression and friction between mole-
cules to release oil. These extraction methods do not use
chemicals. Screw press and hydraulic oil extraction methods
have lower extraction efficiency than solvent extraction. The
disadvantage of solvent extraction is the toxic residual sol-
vents (Cheah et al. 2006; Herrero et al. 2006; Fornari et al.
2012). Therefore, supercritical carbon dioxide extraction
has been applied. Supercritical fluid extraction uses super-
critical carbon dioxide as a solvent. In this extraction
method, carbon dioxide is in an indistinguishable state
between liquid and gas and exhibits gas-like flow behavior
with good penetration capability and liquid-like solvating
ability. This method shows improved extraction efficiency,
selectivity, and environmental benefits compared with con-
ventional extraction methods. In addition, this method
of oil extraction requires less time than other methods
(Herrero et al. 2006). Extraction using a supercritical fluid
consists of two steps. First, extraction is performed by main-
taining the temperature and pressure of the fluid above the
critical point. Second, the separation step changes the phase
of the supercritical fluid to the gas phase by depressurization
to release the extract with no chemical or toxic residues
(Lang and Wai 2001; Cheah et al. 2006; Herrero et al. 2006;
Fornari et al. 2012; Chitra et al. 2015; Martin et al. 2015).
The parameters that affect the extraction efficiency include
the characteristics of the sample preparation—such as the
sample size, homogeneity of the sample, and water/moisture
content—and extraction parameters—such as the pressure,
temperature, flow rate, and extraction time. In general,
smaller sample sizes increase the surface area for extraction
and facilitate penetration of supercritical carbon dioxide
into the food matrix (Lang and Wai 2001; Cheah et al.
2 of 12 Journal of Food Processing and Preservation 2017; 41: e13122; V
S. AKKARACHANEEYAKORN ET AL.
2006). Moisture in the sample affects the extraction process.
Water improves the permeability of the plant cell mem-
brane, causes plant matrix swelling, and aids the flow of ana-
lytes through the matrix. However, >5% w/w of moisture in
the sample may cause poor supercritical fluid and analyte
contact. The effects of pressure and temperature are as fol-
lows: at a constant temperature, increasing the pressure
increases the density and solvating power of supercritical
fluid, thus improving the extraction of the analyte. At a con-
stant pressure, increasing the temperature increases the sol-
vating power of supercritical fluid. However, high
extraction temperatures may reduce the quantity of the ana-
lyte due to thermal degradation.
There are many reports in the literature of using super-
critical carbon dioxide for extraction of high-value phyto-
chemicals from herbs and other plants, i.e., flax, solin,
pomegranate, hyssop, canola, and gac (Lang and Wai 2001;
Barthet and Daun 2002; Abbasi et al. 2008; Kazazi and
Rezaei 2009; Pederssetti et al. 2011; Martins et al. 2015).
Some studies have attempted to develop models to acquire
useful information on the mechanisms and optimization of
the supercritical carbon dioxide extraction process process
(Lang and Wai 2001; Pederssetti et al. 2011; Tai and Kim
2014; Martins et al. 2015). Martins et al. (2015) studied the
optimal conditions for the extraction of aril oil from gac
(Momordica cochinchinensis Spreng) using supercritical car-
bon dioxide to identify the conditions that would produce
the highest percentage of products at the lowest cost. They
varied the pressure from 200 to 400 bar and temperature
from 40 to 60C and used an extraction time of 1 h. They
found that the optimization process to minimize the manu-
facturing cost (8 euro per kilogram of extracted oil) was a
pressure of 400 bar, a temperature of 60C and an extraction
time of 1 h. Tai and Kim (2014) studied the aril oil extrac-
tion process of gac (Momordica cochinchinensis Spreng) and
considered the conditions that produced the highest per-
centage of product and the amount of carotene by varying
pressure from 200 to 400 bar, temperature from 40 to 70C,
and specific flow rate from 50 to 90 kg h21 carbon dioxide
per kg gac aril. They found that the optimal conditions were
a specific flow rate of 70 kg h21 carbon dioxide per kg gac
aril, pressure of 400 bar, temperature of 70C and extraction
time of 2 h. The carotene concentration in the extracted gac
oil was 11,000 ppm.
From related research, no studies have determined the
optimal conditions for the extraction of aril oil from
Momordica cochinchinensis Spreng using supercritical car-
bon dioxide that produce the highest extraction efficiency
and maintain the highest amounts of b-carotene, lycopene,
iodine value, and the lowest levels of acidity and peroxide.
Therefore, this research aimed to determine these optimal
conditions, as well as mathematical equations that can pre-
dict the percentage of extraction efficiency and the amounts
C 2016 Wiley Periodicals, Inc.
S. AKKARACHANEEYAKORN ET AL. EXTRACTION OF ARIL OIL FROM GAC

TABLE 1. FACTORS AND LEVELS OF EACH FACTOR IN THE
EXPERIMENTS
Level
Factor 21.414 21 0 11
Temperature (C) 31 35 45 55
Pressure (bar) 130 150 200 250
of b-carotene, lycopene, iodine value, acidity, and peroxide
using the CCD method for factors of temperature and pres-
sure. The accuracy of the mathematical models was vali-
dated through comparison with the experimental results.
The physical properties of the aril oil extract from Momord-
ica cochinchinensis Spreng, such as color, viscosity, and con-
sumers’ acceptance of its smell, resulting from the optimal
conditions were also studied. Finally, the quality of oil
extracts from the supercritical fluid and screw press meth-
ods was compared.
EXPERIMENTAL METHOD
Aril oil was extracted from Momordica cochinchinensis
Spreng using supercritical carbon dioxide. Ripe gac fruits
were collected from the Nakornpathom Province in the cen-
tral region of Thailand 6 days after harvest (fully ripe; red
color). The gac aril was first dried using a microwave oven
to improve the oil extraction efficiency. Kha et al. (2013b)
compared two methods for drying aril oil from Momordica
cochinchinensis Spreng, microwave and hot air oven. They
found that using hot air resulted in 25% lower oil extraction
efficiency than using a microwave. In addition, using hot air
also caused greater loss of nutritional value than using a
microwave. Therefore, microwave drying was chosen for the
present research. After the gac aril was dried, it was steamed
and extracted using a supercritical carbon dioxide fluid as a
solvent because it is readily available, non-toxic, inexpen-
sive, non-flammable, does not stain the equipment, and
11.414 does not affect the environment. Carbon dioxide is a good
solvent for non-polar compounds; its critical temperature
60
(Tc) and pressure (Pc) are 31.1C and 73 atm, respectively,
271
which allow easily elimination of carbon dioxide from the
extract by leaving it at room temperature.
Sample Preparation
Drying and grounding process. To dry the gac aril, the
method presented by Kha et al. (2013a,b) was used, with
slight modifications. First, 200 g of fresh gac aril was dried
using a microwave (MW 7803, Severin, Germany) at 720 W
for 20 6 1 min. Then, 15 g of dried gac aril was blended with
a blender (Model T30, Moulinex, China) for 15 min. The
ground gac aril was stored away from sunlight at room tem-
perature in a zipper storage bag to avoid any increase in
moisture.
Steaming process. The ground gac aril was steamed
using the method of Kha et al. (2013a,b). Five hundred
grams of ground gac aril were placed on a sheet of white
cloth and spread out in a layer that was 5-mm thick. Then,
the ground gac aril was steamed in a 20-cm-diameter cooker
using 200 mL of water for 20 min, allowed to cool and
stored in a zipper storage bag away from sunlight at room
temperature.
Oil extraction. Oil extraction using supercritical carbon
dioxide was performed on 300 g of ground gac aril using a
supercritical carbon dioxide extraction device (24L-SFE,
Guangzhou Masson New Separation Technology, China).
Carbon dioxide (99.5% w/w) was pumped and then heated

TABLE 2. THE CENTRAL COMPOSITE DESIGN WITH SIX INDEPENDENT VARIABLES FOR OIL EXTRACTION FROM GAC ARIL BY SUPERCRITICAL
CARBON DIOXIDE
b-carotene Lycopene Acidity Peroxide
Temperature (C), Pressure (bar), Extraction content* content* Iodine value* value*
Exp. no. (coded value) (coded value) efficiency (%) (mg/100 g) (mg/100 g) value* (%) (mg KOH/g) (meq/kg)
1 35, (21) 150, (21) 28.71 11.98 4.16 61.43 6 1.31 3.13 6 0.32 N/A
2 60, (11.414) 200, (0) 81.42 34.58 12.42 71.49 6 3.07 2.85 6 0.29 N/A
3 31, (21.414) 200, (0) 87.57 19.79 6.26 60.87 6 3.90 2.09 6 0.42 N/A
4 55, (11) 150, (21) 28.14 30.64 10.72 62.37 6 1.40 3.04 6 0.19 N/A
5 45, (0) 200, (0) 72.96 25.97 7.97 62.66 6 0.57 1.80 6 0.21 N/A
6 45, (0) 200, (0) 95.43 28.59 8.90 62.48 6 2.96 1.82 6 0.33 N/A
7 55, (11) 250, (11) 100.00 43.04 13.53 64.29 6 2.73 2.16 6 0.29 N/A
8 35, (21) 250, (11) 93.82 17.65 5.56 70.97 6 2.32 0.78 6 0.23 N/A
9 45, (0) 130, (21.414) 16.59 29.48 9.09 59.27 6 3.37 4.68 6 0.21 N/A
10 45, (0) 200, (0) 78.71 29.41 7.33 67.50 6 1.27 1.47 6 0.16 N/A
11 45, (0) 271, (11.414) 100.00 28.25 10.95 69.62 6 3.10 4.14 6 0.49 N/A

Note: *Means 6 sd, n 5 3.

N/A indicates not detected.

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EXTRACTION OF ARIL OIL FROM GAC S. AKKARACHANEEYAKORN ET AL.

TABLE 3. MODELS, R2 AND %D OF THE OIL EXTRACTION EFFICIENCY, B-CAROTENE AND LYCOPENE CONTENTS, IODINE VALUE, AND ACIDITY
VALUE IN CODED UNITS
P value of the
Y Models R2 (%) lack of fit Ypredict Yexperimental %D
2
Oil extraction 82.3686 - 0.3867x1 1 31.8651x2 – 1.12(x1) 94.93 0.698 92.28 97.50 5.35
efficiency (%) – 14.2181(x2)2 1 1.69x1x2
b-carotene content 27.9919 1 8.1194x1 1 2.0407x2 – 0.9534(x1)2 81.38 0.072 36.48 36.05 6.19
(mg/100 g) – 0.1140(x2)2 1 1.6836x1x2
Lycopene content 8.0629 1 2.9056x11 89.90 0.209 11.79 11.01 7.08
(mg/100 g) 0.8558x2 1 0.3413(x1)2 1 0.6811(x2)2 1 0.3509x1x2
Iodine value (%) 64.2114 1 1.1579x11 3.2626x21 0.8466(x1)2 61.49 0.319 66.69 67.55 1.27
– 0.0227(x2)2 - 1.9040x1x2
Acidity value 1.6770 1 0.2928x1 – 0.5094x2 1 0.1028(x1)2 74.23 0.033 2.19 2.32 5.60
(mg KOH/g) 1 1.0843(x2)2 1 0.3673x1x2

Note:
x1 5codevalueoftemperature ð21:414  x1  1:414Þ.
x2 5codevalueofpressure ð21:414  x2  1:414Þ.
absðYpredict 2Yexperimental Þ3100%
%D5 Yexperimental .

to the desired extraction pressure and temperature, which Analyses

were 31, 35, 45, 55, and 60C and 130, 150, 200, 250, and
271 bar for 2 h. Before pumping, precooling of the solvent
was generally required to avoid pump cavitation. The
supercritical solvent was continuously loaded from the bot-
tom of the extraction vessel. At the exit of the extractor, the
supercritical solvent with gac aril oil extract flowed to two
separators in series; because of the lower pressure, the
extracted gac aril oil was separated from the gaseous solvent
and collected.
The obtained gac aril oil was stored in a completely
sealed bottle in the dark at room temperature and at 5C.
The percentage of extraction efficiency was determined,
and chemical analysis was performed to determine the
iodine value, acidity, peroxide, b-carotene, and lycopene.
Physical analysis was performed to determine the color
and viscosity. The acceptance of its smell by consumers was
also analyzed.
Extraction efficiency percentage. The efficiency of the
oil extraction was derived from the ratio of the weight of the
obtained oil extract after subtracting the moisture content
and the weight of the oil extract, as determined using a
Soxhlet extractor.
weightof oilextract2moisturecontent
Extractionefficiency5
weightof oilextractusingaSoxhletextractor
The moisture content was measured using the Ca 2e-55
method (AOCS 1997).
The total oil from the gac fruit was treated with a micro-
wave before extraction and was then analyzed using a BUCHI €
€
extraction system (B-8118, BUCHI, Switzerland) by weighing
3 g of the gac aril in a cellulose thimble and placing it in the
extractor. Filtered petroleum ether (boiling point 60–90C)
was added to the weighed aluminum vessel, which was then
connected to the extraction system. The temperature used
was 145C, and the extraction time was 120 min; the time
required to wash out the remaining fatty acids was 20 min,
and the aluminum vessel used for storing the oil extract was
dried for 20 min. The sample was dried in an oven at 100C
for 30 min until the weight was constant. The amount of oil
in the sample was then used to calculate the weight.
Iodine values were measured according to AOCS using
the 1d-92 method (1997); the acidity values were measured
according to the AOCS Cd 3d-63 method (1997); and the
peroxide values were measured according to the AOCS Cd
8b-90 method (1997).

FIG. 1. THE 3D GRAPH SHOWING EXTRACTION EFFICIENCY OF ARIL b-Carotene and lycopene contents. The method for
OIL FROM MOMORDICA COCHINCHINENSIS SPRENG AT VARIOUS determining the b-carotene and lycopene contents was
TEMPERATURES (X1) AND PRESSURES (X2) modified from Nagata and Yamashita (1992). A 0.1 g

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S. AKKARACHANEEYAKORN ET AL.
FIG. 2. THE 3D GRAPH SHOWING THE AMOUNT OF b-CAROTENE IN
OIL FROM GAC ARIL OF MOMORDICA COCHINCHINENSIS SPRENG AT
VARIOUS TEMPERATURES (X1) AND PRESSURES (X2)
sample of oil was added to a mixture of acetone:hexane, 4:6 (v/
v), and was vortexed for 1 min. It was then diluted by a factor
of 10, and the light absorption of the solution was subse-
quently determined on a UV–visible spectrophotometer at
wavelengths of 663, 645, 505, and 453 nm, using an acetone:-
hexane solution at a 4:6 (v/v) ratio as a blank. The values were
used to calculate the lycopene content with Eq. 1 and the b-
carotene content with Eq. 2.
Lycopeneðmg=100gfreshweight;FWÞ520:0458A663
(1)
10:204A645 10:372A505 20:0806A453
b2caroteneðmg=100gfreshweight;FWÞ50:216A663
(2)
21:220A645 10:304A505 20:452A453
Color value. A 2-mL oil sample from the gac fruit was
added to a sample container for color measurement. The
measurement was performed in triplicate using a HunterLab
ColorQuest 450 (Color Global, Reston, VA) in a CIE lab
system.
FIG. 3. THE 3D GRAPH SHOWING THE AMOUNT OF LYCOPENE IN OIL
FROM GAC ARIL OF MOMORDICA COCHINCHINENSIS SPRENG AT
VARIOUS TEMPERATURES (X1) And Pressures (X2)
Journal of Food Processing and Preservation 2017; 41: e13122; V
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EXTRACTION OF ARIL OIL FROM GAC
Apparent viscosity. The apparent viscosity was meas-
ured on 15 mL of oil using a Brookfield viscometer (RVDV-
II, Brookfield, Middleboro, MA) with the No. 27 spindle at
100 rpm and room temperature.
Acceptance of smell by consumers. The acceptance of
smell by consumers was studied using a nine-point hedonic
scale with 20 untrained panelists.
Statistical Analysis
Determining the optimal conditions and equations for pre-
dicting the percentage of extraction efficiency and the
amounts of b-carotene, lycopene, iodine value, acidity, and
peroxide in the extraction of gac aril oil using supercritical
carbon dioxide was planned using response surface method-
ology (RSM) with the CCD method. Temperature and pres-
sure were selected as independent variables. Each
independent variables had five levels, as shown in Table 1,
and the middle levels were repeated twice, as shown in Table
2. There were six response values (Y): the percentage of
extraction efficiency and the amounts of b-carotene, lyco-
pene, iodine value, acidity, and peroxide. Each value was
measured three times. Results were fitted into the empirical
second order polynomial model. Data analysis was per-
formed using Minitab software version 16.2.2.0 at the 95%
confidential level.
To compare the extraction efficiency, chemical properties,
such as the amounts of b-carotene, lycopene, iodine value,
acidity, and peroxide, and physical properties, such as the
color (L*, a*, b*, and h*) and viscosity, were analyzed using
one-way ANOVA at a 95% confidential level in Minitab soft-
ware version 16.2.2.0, and the acceptance of smell by con-
sumers was determined using two-sided, two alternative
forced choice (2-AFC) at a 95% confidential level on gac aril
oil products extracted using screw press and supercritical
fluid methods.
RESULTS AND DISCUSSION
Table 2 shows the results of determining the optimal condi-
tions planned using the CCD method for two factors and
using the response surface to find the optimal temperature
and pressure for the extraction of aril oil from Momordica
cochinchinensis Spreng to maximize the extraction efficiency
and the amounts of b-carotene, lycopene, and iodine value
and to minimize the acidity.
Analysis of the Extraction Efficiency of Aril
Oil from Momordica cochinchinensis Spreng
Under Various Conditions
The observed responses from Table 2 were used to compute
the model coefficients using the least square method. Table
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EXTRACTION OF ARIL OIL FROM GAC
FIG. 4. THE 3D GRAPH SHOWING THE IODINE VALUE IN OIL FROM
GAC ARIL OF MOMORDICA COCHINCHINENSIS SPRENG AT VARIOUS
TEMPERATURES (X1) AND PRESSURES (X2)
3 shows the prediction of the extraction efficiency of aril oil
from Momordica cochinchinensis Spreng with an R2 of 94.93,
indicating that the independent variables (temperature and
pressure) can explain the variation or change in the depend-
ent variable (extraction efficiency) and that the quadratic
model can be used to create an equation that accurately and
appropriately predicts the response.
Considering the lack of fit of the equation, the P value of
the lack of fit was 0.698, which is higher than 0.05, indicat-
ing that it was not significantly different. This means that
the established equation does not have a lack of fit or that
the equation fit well with the results, indicating that any
independent variable (temperature and pressure) can be
used in the equation to explain or predict the value of the
dependent variable (extraction efficiency). This includes
R2, which was 94.93%, indicating that the quadratic model
can be used to create an equation that accurately and suit-
ably explains or predicts the value of the dependent vari-
able. Therefore, the above equation can be used for the
prediction of the extraction efficiency of gac aril oil from
Momordica cochinchinensis Spreng under range operation
conditions.
FIG. 5. THE 3D GRAPH SHOWING THE ACIDITY OF OIL FROM THE ARIL
OF MOMORDICA COCHINCHINENSIS SPRENG AT VARIOUS
TEMPERATURES (X1) AND PRESSURES (X2)
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S. AKKARACHANEEYAKORN ET AL.
After obtaining the equation to predict the results, a sur-
face graph of the extraction efficiency results was plotted, as
shown in Fig. 1. Figure 1 shows that pressure is the key
parameter that significantly influences the extraction effi-
ciency. From the results, it can be observed that increasing
the pressure at a constant temperature increased oil extrac-
tion efficiency. Increasing the pressure increased the density
and solvating power due to greater attractive forces between
supercritical carbon dioxide and solute molecules, which
also increased the oil extraction efficiency (McHugh et al.
1994; Tonthubthimthong et al. 2001; Cheah, et al. 2006).
When the temperature increased at pressures of 150 and 200
bar, the extraction efficiency decreased because increasing
the temperature caused the density of carbon dioxide to
decrease, reducing the extraction efficiency. However,
increasing the temperature did not have an obvious effect
on the extraction efficiency because increasing the tempera-
ture increased the vapor pressure of the oil, causing the oil
to dissolve better in supercritical carbon dioxide. Mean-
while, the density of the supercritical carbon dioxide also
decreased, causing the solubility by carbon dioxide to
decrease as well. Nevertheless, the optimal temperature and
pressure also depends on the samples extracted.
b-Carotene and Lycopene Analysis of the Oil
from Arils of Momordica cochinchinensis
Spreng Under Various Conditions
The highest amounts of b-carotene and lycopene of 43.04
and 13.53 mg/100 g, respectively, were obtained at 55C and
250 bar in experiment number 7, and the lowest amounts of
b-carotene and lycopene of 11.98 and 4.16 mg/100 g,
respectively, were obtained at 35C and 150 bar in experi-
ment number 1, as shown in Table 2.
Analysis of the coefficient of determination and variance
from regression analysis of the response surface of b-
carotene and lycopene in the oil revealed an R2 of 81.38,
indicating that the independent variables (temperature and
pressure) can explain the variation or change in the depend-
ent variable (the amount of b-carotene). This also means
that the second order polynomial model can be used to cre-
ate an equation that accurately and appropriately predicts
the response value.
Table 3 shows the creation of the equation to predict the
amount of b-carotene in the gac aril oil using the factor val-
ues from the analysis of the coefficient of regression equa-
tion for the amount of b-carotene in the oil.
Considering the lack of fit of the equation, the P value of
the lack of fit was 0.072, which is higher than 0.05 and,
therefore, not significantly different (P > 0.05). This either
means that the created equation does not have a lack of fit
or the obtained equation fit the results and any independent
variable (temperature and pressure) can explain or predict
C 2016 Wiley Periodicals, Inc.
S. AKKARACHANEEYAKORN ET AL.
FIG. 6. DETERMINING THE OPTIMAL
EXTRACTION CONDITIONS FOR GAC ARIL
OIL FROM MOMORDICA
COCHINCHINENSIS SPRENG BY
SUPERCRITICAL FLUID EXTRACTION
the value of the dependent variable (the amount of b-caro-
tene). Therefore, this equation is suitable for use. The R2
was 81.38%, indicating that the quadratic model can be
used to create an equation that accurately and appropriately
predicts the response value. Therefore, the above equation
can be used to predict the amount of b-carotene in oil from
the aril of Momordica cochinchinensis Spreng.
After obtaining the prediction equation, the amount of
b-carotene in the gac aril oil was plotted using response sur-
face graphs, as shown in Fig. 2. Figure 2 shows that tempera-
ture and pressure are the crucial parameters that influence
the amount of b-carotene. From the results, it can be
observed that increasing the temperature at high pressure
increased the amount of b-carotene in the oil extracted
from gac aril of Momordica cochinchinensis Spreng. Increas-
ing the temperature at high pressure of supercritical carbon
dioxide increased the volatility and enhanced the solvating
power, thereby increasing the amount of b-carotene. Tem-
perature directly affects the desorption, diffusion and disso-
lution of the analyte (Lang and Wai 2001; Cheah, et al.
2006). Figure 2 shows that at a constant temperature,
increasing the pressure increased the amount of b-carotene.
This result is consistent with the results of Tai and Kim
(2014). Increasing the pressure increased the solvating
power, and more diffusion in the plant matrix led to an
increased amount of b-carotene.
Lycopene Analysis of Gac Aril Oil under
Various Conditions
From the regression analysis of the response surface of the
amount of lycopene in the oil, the R2 was 89.90%, indicating
that the independent variables (temperature and pressure)
can explain the variation or change in the dependent vari-
able (the amount of lycopene). This means that the quad-
Journal of Food Processing and Preservation 2017; 41: e13122; V
C 2016 Wiley Periodicals, Inc.
EXTRACTION OF ARIL OIL FROM GAC
ratic model can be used to create an equation that accurately
and appropriately predicts the response value.
Table 3 shows the creation of an equation to predict the
amount of lycopene in oil from gac arils by taking the values
of factors from the analysis of the coefficient of regression
equation of the lycopene content.
Considering the lack of fit of the equation, the P value of
the lack of fit was 0.209, which is higher than 0.05 and,
therefore, not significantly different (P > 0.05). This means
that the created equation does not have lack of fit with the
results and that any independent variable (temperature and
pressure) can explain or predict the value of the dependent
variable (the amount of lycopene). Therefore, this equation
is suitable for use. In addition, the R2 was 89.90%, indicating
that the quadratic model can be used to create an equation
that accurately and appropriately predicts the response
value. Therefore, the above equation can be used to predict
the amount of lycopene in oil from the aril of Momordica
cochinchinensis Spreng.
After obtaining the prediction equation, the lycopene
content in the oil was plotted using response surface graphs,
as shown in Fig. 3. Figure 3 shows that temperature and
pressure are the crucial parameters that influence the
amount of lycopene. The amount of lycopene in the oil
extracted from the gac aril of Momordica cochinchinensis
Spreng increased linearly with temperature and pressure.
Iodine Analysis of Oil from the Aril of
Momordica cochinchinensis Spreng
The iodine value in the extracted oil using supercritical fluid
ranged from 59.27 to 71.49%, as shown in Table 2, which is
consistent with the research by Kha et al. (2013b). These
authors found that gac arils contained 70% saturated fatty
acids out of all fatty acids and that 50% of all unsaturated
fatty acids contained polyunsaturated fatty acids. Our values
7 of 12
EXTRACTION OF ARIL OIL FROM GAC
FIG. 7. DETERMINING THE OPTIMAL CONDITIONS FOR THE
EXTRACTION OF GAC ARIL OIL
are also consistent with the research by Betty et al. (2004),
who found that arils of Momordica cochinchinensis Spreng
consisted of the following fatty acids by weight: 32% oleic
acid, 29% palmitic acid, and 28% linoleic acid. Therefore,
oil from the arils of Momordica cochinchinensis Spreng con-
tains high amounts of unsaturated fatty acids.
Table 3 shows the creation of a second order polynomial
equation to predict the values of iodine in gac aril oil by tak-
TABLE 4. THE EXTRACTION EFFICIENCY, AMOUNT OF CAROTENE AND CHEMICAL PROPERTIES OF GAC ARIL OIL EXTRACTED USING
SUPERCRITICAL CARBON DIOXIDE AND A SCREW PRESS
Extraction
Extraction method efficiency (%)
Supercritical fluid (this study) 97.50*
Screw press (Akkarachaneeyakorn et al. 2015) 61.82*
Statistical relationship of extraction methods * *
Note: *Significant difference in the same column (P  0.05).
*Significant at P  0.05.
N.S. Non-significant at P > 0.05.
8 of 12
S. AKKARACHANEEYAKORN ET AL.
ing the values of the factors from the analysis of the coeffi-
cient of regression equation of the iodine value in the oil.
From the regression analysis of the iodine value, the R2
was 61.49%, and the P value of the lack of fit of the equation
was 0.319, which is higher than 0.05 and, therefore, not sig-
nificantly different (P > 0.05). This either means that the
created equation does not have a lack of fit or that the equa-
tion fits the results and any independent variable (tempera-
ture and pressure) in the equation can explain or predict the
value of the dependent variable (the iodine value). There-
fore, this equation is suitable for use and can be used to pre-
dict the iodine value in oil from gac arils.
After obtaining the prediction equation, the iodine value
of the oil was plotted using response surface graphs, as
shown in Fig. 4. Figure 4 shows that temperature and pres-
sure are the crucial parameters that influence the amount of
iodine. The iodine concentration in the oil extracted from
the gac aril of Momordica cochinchinensis Spreng increased
when the temperature and pressure increased. However, it
was observed that at pressure of 250 bar, the amount of
iodine decreased with increasing temperature.
Acidity Analysis
The acidity of the oil from gac arils was 0.78–4.68 mg potas-
sium hydroxide per gram, as shown in Table 2. The Notifica-
tion of the Ministry of Public Health, Thailand No. 205 B.E.
2543 (2000), specified that the acidity shall not exceed
4.00 mg potassium hydroxide per gram.
Creating a second order polynomial equation to predict
the acidity of oil from gac arils by taking factor values from
the analysis of the coefficient of regression equation for the
acidity of the oil is shown in Table 3.
From the regression analysis of the oil’s acidity, the R2 was
74.23%, indicating that the model can be used to accurately
and appropriately predict the values of the dependent variables.
Therefore, the above equation can be used to predict the acidity
of oil from the arils of Momordica cochinchinensis Spreng.
After obtaining the prediction equation, the oil acidity
was plotted using response surface graphs, as shown in
Fig. 5. Figure 5 shows that temperature and pressure are the
b-carotene Lycopene Iodine Acidity Peroxide
(mg/100 g) (mg/100 g) value (%) (mg KOH/g) (meq/kg)
36.05* 11.01 67.55* 2.32* N/A
18.22* 5.02 63.42* 3.92* N/A
N.S. * * –
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S. AKKARACHANEEYAKORN ET AL. EXTRACTION OF ARIL OIL FROM GAC

TABLE 5. PHYSICAL PROPERTIES OF THE ARIL OIL FROM MOMORDICA COCHINCHINENSIS SPRENG EXTRACTED USING SUPERCRITICAL
CARBON DIOXIDE AND A SCREW PRESS
Apparent Smell**
Extraction method L* a* b* h* (8) viscosity (cP) (n 5 20 panelists)
Supercritical fluid (this study) 12.18* 26.91* 13.60 26.67 97.20 6 3.45 10
Screw press (Akkarachaneeyakorn et al. 2015) 7.67* 21.60* 11.02 25.16 105.75 6 6.66 10
Statistical relationship of extraction methods * * N.S. N.S. N.S. N.S.

Note:*Significant difference in the same column (P  0.05).

**Significant at P  0.05.
N.S. Non-significant at P > 0.05.

crucial parameters that influence the acidity. The acidity of
the oil extracted from the gac aril of Momordica cochinchi-
nensis Spreng increased when the temperature increased and
the pressure was held constant at 250 bar, owing to a hydro-
lysis reaction. High temperature causes reduction of the
activation energy for lipid peroxidation and decomposes
preformed hydroperoxides into free radicals, which stimu-
lates the autoxidation process and leads to elevated oil
acidity.
Peroxide Analysis
Peroxide was not found in the gac aril oil, as shown in Table 2,
because oil from the arils of Momordica cochinchinensis
Spreng contains vitamin E, which helps prevent rancidity in
the oil. This is consistent with the research by Vuong and
King (2003), who found that oil from gac arils contained 334
mg g21 of vitamin E, 69% unsaturated fatty acids, and 35% of
polyunsaturated fatty acids. Furthermore, Vuong et al. (2005)
found that the arils of Momordica cochinchinensis Spreng con-
tained 76.30 6 22.20 mg mL21 of a-tocopherol.
Determining the Optimal Extraction
Conditions for Gac Aril Oil Using
Supercritical Fluid Extraction
Determining the optimal conditions to yield the highest
extraction efficiency, the highest amounts of b-carotene, lyco-
pene, iodine value, and the lowest acidity was performed using
the response optimizer function in Minitab 16.2.2.
Figure 6 shows the temperature and pressure interval that
resulted in an extraction efficiency between 80 and 100%,
30–50 mg of b-carotene per 100 g, 10–20 mg of lycopene
per 100 g, 60–80% iodine value, and an acidity of 0–4 mg
potassium hydroxide. The white area represents the optimal
conditions that produced the highest extraction efficiency,
the highest amounts of b-carotene, lycopene, and iodine
value, and the lowest acidity and peroxide values.
The factors affecting the supercritical fluid extraction were
temperature and pressure because the increase or decrease in
temperature causes the density of the solvent to increase or
decrease, respectively, which affects the amount of compounds
obtained from the extraction and the loss in nutritional value
of the sample. The pressure affects the density of the solvent,
which also affects the solubility due to the solvent at which the
extraction by the fluid near the supercritical phase would have
the optimal pressure. If the pressure is increased beyond this
point, the extraction rate will decrease because, at high pres-
sure, the dispersion ability of the fluid decreases and greatly
compresses the raw materials, reducing the space.
Validation of the Accuracy of the
Mathematical Model of the Extraction Using
Supercritical Fluid
The mathematical model used to predict the extraction coeffi-
cient of the amounts of b-carotene, lycopene, iodine value, and
acidity is shown in Table 3. To confirm the results of the analy-
sis, an experiment at 55C and 220 bar was performed. The pro-
gram calculated an extraction efficiency of 92.28%, an amount
of b-carotene of 36.48 mg/100 g, an amount of lycopene of
11.79 mg/100 g, an iodine value of 66.69%, and an acidity of
2.19 mg potassium hydroxide per gram, as shown in Fig. 7.

TABLE 6. EXTRACTION EFFICIENCY PERCENTAGE FOR OIL EXTRACTION FROM GAC ARILS
Extraction equipment

Drying method Supercritical fluid Screw press Hydraulic

Microwave 97.50 (this study) 61.82 (Akkarachaneeyakorn et al. 2015) 93.00 (Kha et al. 2013b)
Hot air 95.00 (Kha et al. 2014) – 68.00 (Kha et al. 2013b)

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EXTRACTION OF ARIL OIL FROM GAC S. AKKARACHANEEYAKORN ET AL.

From Table 3, the %D values of the extraction efficiency
and the amounts of b-carotene and lycopene, iodine value,
and acidity were all under 10%, indicating that all mathe-
matical models were accurate and appropriate for use.
Comparison of the Extraction of Aril Oil
from Momordica cochinchinensis Spreng
Using the Supercritical Fluid and Screw
Press Methods
Table 4 shows the extraction efficiency and the amounts of
b-carotene, iodine value, and acidity. These values were sig-
nificantly different (P  0.05) for extraction using the super-
critical fluid and screw press methods. Table 5 shows
physical properties of the aril oil from Momordica cochinchi-
nensis Spreng extracted using supercritical carbon dioxide
and a screw press. The L* and a* of the gac aril oil from both
methods were also significantly different (P  0.05). For
lycopene, b* and h* as well as viscosity were not significantly
different (P > 0.05). The similarity of the smell was eval-
uated using 20 untrained panelists, and the difference was
not significant (P > 0.05). Therefore, supercritical fluid
extraction was better than using a screw press in terms of
extraction efficiency, the carotenoid contents, and the
chemical properties because supercritical fluid affords pure
compounds with unwanted chemicals separated out, but a
screw press compresses the sample. The disadvantages of
supercritical fluid extraction involve its expense and the fact
that the solvent used must be suitable for the sample. There-
fore, supercritical fluid extraction is suitable for a medium
industrial level, and screw press extraction is suitable for a
community level and a small industrial level because of its
low cost. However, the extraction using a screw press results
in lower extraction efficiency and a lesser quality of oil
because the extraction temperature cannot be controlled.
The results in Table 6 show that extraction using a super-
critical fluid combined with microwave drying of the gac
arils before extraction resulted in the highest extraction effi-
ciency compared to other studies.
Drying using a microwave resulted in 2.5% better extrac-
tion efficiency than using hot air as shown in Table 6
because drying using a microwave evaporates water from
the cells, causing the cells of the gac aril to break and yield a
greater amount of oil when extracted. Compared with drying
using hot air, which is based on the principle of introducing
heat to the surface to evaporate water, the cells are not dam-
aged. Therefore, the oil is more difficult to remove from cells
than when drying using a microwave (Kha et al. 2013b). This
is consistent with the research of Rama et al. (2010). They
studied the extraction of oil from flaxseeds by comparing
Soxhlet, screw press, and supercritical fluid extraction meth-
ods and found that extraction using a supercritical fluid pro-
duced a greater amount of oil than when using a screw press.
In terms of carotenoid content, this study only measured the
carotenoid content trend using the method of Nagata and
Yamashita (1992) and could not determine the exact amount
of the compound due to limitations of the equipment used for
the measurement. However, the carotenoid content trend was
consistent with the research by Kha et al. (2014) and Martine
et al. (2008), who found that supercritical fluid extraction pro-
duced higher amounts of carotenoids than using a screw press.
In addition, the chemical properties of the gac aril oil
extracted using the supercritical fluid were significantly differ-
ent from the gac aril oil extracted using a screw press
(P  0.05) as shown in Table 4. Moreover, when the amount
of b-carotene and lycopene increased, the iodine value also
increased because iodine will react with the double bonds in
b-carotene and lycopene as the double bonds in unsaturated
fatty acids.
CONCLUSION
Oil Extraction from the Arils of Momordica
cochinchinensis Spreng Using a Supercritical
Fluid
The extraction using supercritical fluid was studied to show
how the temperature and pressure affected the extraction
efficiency, amount of b-carotene, amount of lycopene,
iodine value, and acidity, which resulted in the following
mathematical models.

Oil extraction efficiency 5 82:3686 2 0:3867x1 1 31:8651x2 – 1:12ðx1 Þ2 – 14:2181ðx2 Þ2 11:69x1 x2

b2carotene content 5 27:9919 1 8:1194x1 1 2:0407x2 – 0:9534ðx1 Þ2 – 0:1140ðx2 Þ2 11:6836x1 x2
Lycopene content 5 8:0629 1 2:9056x1 1 0:8558x2 1 0:3413ðx1 Þ2 1 0:6811ðx2 Þ2 10:3509x1 x2
Iodine value 5 64:2114 1 1:1579x1 1 3:2626x2
Acidity value 5 1:6770 1 0:2928x1 – 0:5094x2 1 0:1028ðx1 Þ2 1 1:0843ðx2 Þ2 10:3673x1 x2

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S. AKKARACHANEEYAKORN ET AL.
Where:
x1 5codevalueof temperature ð21:414  x1  1:414Þ;
x2 5codevalueof pressure ð21:414  x2  1:414Þ
The Optimal Conditions for the Extraction
of Gac Aril Oil
The optimal conditions for the extraction of aril oil from
Momordica cochinchinensis Spreng using a supercritical fluid
were 52–55C and 200–250 bar, which resulted in an extrac-
tion efficiency of 80–100%, 30–50 mg b-carotene per 100 g,
10–20 mg lycopene per 100 g, 60–80% iodine values, and an
acidity of 0–4 mg potassium hydroxide per gram.
Comparison of Gac Aril Oil Extracts
Extracted from the Supercritical Fluid and
Screw Press Methods
Supercritical fluid extraction resulted in a better extraction
efficiency of oil with higher amounts and better chemical
properties of carotenoids compared to using a screw press.
ACKNOWLEDGMENT
The authors are grateful to the King Mongkut’s University
of Technology North Bangkok for funding the research
(Contract no. KMUTNB-GEN-58-23).
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