Separation and Purification Technology 141 (2015) 354–365

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Separation and Purification Technology

journal homepage: www.elsevier.com/locate/seppur

Modeling and response surface analysis of supercritical extraction

of watermelon seed oil using carbon dioxide
Amit Rai a,⇑, Bikash Mohanty b, Ravindra Bhargava b
a
Department of Chemical Engineering, NIT Hamirpur, Hamirpur, Himachal Pradesh 177001, India
b
Department of Chemical Engineering, IIT Roorkee, Roorkee, Uttarakhand 247667, India

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

Article history: Supercritical fluid extraction with supercritical CO2 (SC-CO2) is applied to extract the vegetable oil from
Received 16 October 2014 watermelon seeds. A small face central composite design (SFCCD) is employed to optimize the extraction
Received in revised form 2 December 2014 parameters. The maximum value of cumulative extraction yield (CEY) for watermelon oil is found to be
Accepted 5 December 2014
about 51.83 wt%, and was obtained when SC-CO2 extraction is carried out at 60 °C, 400 bar, 15 g/min sol-
Available online 23 December 2014
vent flow with 10% co-solvent and with 0.50 mm particle diameter. The analysis of response surface
showed that, extracted data are adequately fitted to reduced second-order polynomial model and all
Keywords:
independent variables, quadratics of independent variables and some interaction parameters between
Watermelon
Response surface methodology
independent variables have significant effects on CEY. The correlation coefficients ‘‘R-square’’, ‘‘adjusted
Seed oil R-square’’ and ‘‘predicted R-square’’ values for reduced quadratic model are 0.9974, 0.9907 and 0.9226,
Supercritical fluid extraction respectively. The chemical composition of seed oil shows that 75.1% unsaturated and 24.9% saturated
fatty acids are present in the seed oil. Further, it is found that watermelon seed is a high source of linoleic
acid.
Ó 2014 Elsevier B.V. All rights reserved.

1. Introduction
Watermelon seeds contain many important functional compo-
nents and are traditionally used for herbal, therapeutic as well as
clinical applications [1] and its oil is extremely suitable emollient
in natural baby care formulations and light body emulsions due
to its stable shelf life and moisturizing capabilities [2]. Watermelon
seed oil has strong antioxidant properties [3] and has been recog-
nized for several health benefits such as prevention of the chronic
diseases such as diabetes, cardiovascular diseases, improving gut
health as well as overweight treatment [2,4]. Further, watermelon
seeds have excellent potential for their use in food formulations,
due to high contents of oil (50–51%) and protein (32–37%) [3–7].
Watermelon seed oil, contains highly unsaturated fatty acids
(77–82%) due to the presence of linoleic acid (59–67.5%) and oleic
acid (14.0–18.1%) and has been used as cooking oil and food addi-
tive in Western countries as well as in the Middle East of Africa
[3–7]. Besides being used as a fruit, watermelon is also used for
preparation of juice, jellies, jams, sauces and salads which produce
large quantities of seeds as a waste. Thus, it offers large potential to
⇑ Corresponding author. Tel.: +91 75790 75744; fax: +91 1332 276535.
E-mail address: mit.rai123456@gmail.com (A. Rai).
use these waste seeds for the production of oil and functional
ingredients.
Conventionally, the vegetable oil is extracted by the means of
mechanical operation such as pressing or by using organic solvent
extraction, such as hexane, petroleum ether, and chloroform [8].
The mechanical device such as press (cold or hot) gives good qual-
ity of oil with low yield [8] whereas organic solvent gives higher
yield with some traces of organic solvents. However, increased
public health concern, rigid environmental and safety regulations
are forcing the industry to search for alternatives to the organic
solvents as traces of residual toxic organic solvents remain with
the oil [9]. In recent years, supercritical extraction of the vegetable
oil has received considerable attention as a promising alternative
to conventional solvent extraction and mechanical pressing pro-
cess. The most commonly used solvent is supercritical carbon diox-
ide because of its unique properties such as, non-polar, non-toxic,
non-flammable, non-explosive, cost efficient, easy to remove and
also offers a higher extraction rate [10].
Under the above backdrop, the present work is focused on
experimental studies of extraction of oil from watermelon seeds
obtained from the northern part of India. The main objective of
the present work was to develop the statistical model between
input parameters such as pressure, temperature, particle diameter,
solvent flow rate, co-solvent addition, with that of the output
parameter (extraction yield). Response surface methodology

http://dx.doi.org/10.1016/j.seppur.2014.12.016
1383-5866/Ó 2014 Elsevier B.V. All rights reserved.
 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365
(RSM) with centered composite design has been used to optimize
the number of experiments. The experimental extraction yield
obtained for different combination of input parameters, as sug-
gested by central composite design, has been interpreted by
response surface methodology. Some important properties of seed
material such as, thermal degradation, functional groups and struc-
ture of seed particles are evaluated by thermo gravimetric analysis
(TGA), Fourier transform infrared (FTIR) spectroscopy and scanning
electron microscopy (SEM) respectively. The free fatty acid present
in the extracted oil has been analyzed by gas chromatography.
2. Experimental
2.1. Materials
The shelled and dried watermelon seeds have been purchased
from local market. The shelled seed material has been unshelled
manually, dried under sunlight and stored at 4 °C. Without any
pre-treatment all the seeds have been chopped in domestic mixer
grinder (Bajaj, India). The particles of chopped seeds have been
separated and graded according to their particle size by certified
test sieves (Endecotts Ltd., London, England) with a vibratory sieve
shaker (octagon 200, Endecotts Ltd., London, England).
The seed particles have been collected in three ranges as shown
in Table 1 and for convenience, mean particle diameter, based on
sieve openings, has been assigned to a fraction between two suc-
cessive sieves. Carbon dioxide used in the experiments is of purity
99.9% and is obtained in pressurized deep tube cylinders supplied
by Sigma gases, India. The ethanol is used as a co-solvent. The
entire standard chemicals used for the analysis are the analytical
and HPLC grade. FAME 37 Mix is obtained from Sigma Chemicals
(St. Louis, MO, USA). Ethanol is purchased from Merck Ltd. (Darms-
tadt, Germany), and toluene, sulfuric acid (98.5%), sodium chloride,
n-hexane, methanol, potassium bicarbonate, anhydrous sodium
sulfate in the form of powder, have been purchased from Merck
Ltd. (Mumbai, India). All the solutions were prepared with ultra-
pure water obtained from Milli-Q system supplied by Millipore,
Bedford, MA, USA.
2.2. Supercritical fluid extraction
A supercritical extraction system, SFE 1000F, supplies by Thar
Technologies Inc., Pittsburgh, has been used for the extraction of
watermelon seed oil using supercritical carbon dioxide above crit-
ical conditions. The extraction system consists of a 1 l extraction
vessel and two separators. A cylindrical stainless steel basket
19 cm high and 7.5 cm in diameter has been used for easy loading
and unloading of the extraction vessel. The cylindrical basket has
been packed with 5 mm diameter glass beads. Initially 150 g glass
beads (3.0 cm height) have been placed at the bottom of the basket,
and then glass wool having a thickness of 0.5 cm is positioned over
it followed by 150 g of glass beads over the glass wool. This
arrangement has been helpful for uniform distribution of super-
critical CO2. Above this arrangement, a layer of 50 g of seed parti-
cles is placed followed by glass beads, glass wool and again glass
beads of heights 3.0, 0.5 & 3.0 cm respectively. This arrangement
stops the carryover of solute particles with supercritical CO2. The
Table 1
Size specification of seed fractions.
No. Sieve opening (mm) Average particle size (mm)
1 1.180 + 0.850 1.00
2 0.850 + 0.600 0.75
3 0.600 + 0.425 0.50
355
extracted oil is collected from the sample points located at the bot-
tom of the separator in every five minute interval in the separate
sample vials. Ethanol, the co-solvent, from the collected extracts
is removed by rotary vacuum evaporator and then remaining oil
samples are measured gravimetrically. The total cumulative
extraction yield is then determined from the sum of all oil samples
extracted. This procedure has been adopted for each experiment.
The operating method of the supercritical extraction unit is
shown in Fig. 1. The liquid CO2 supply from cylinder [a], is passed
through a 0.22 lm sintered stainless steel metal filter to a high
pressure pump [c]. Before reaching the high pressure pump, it is
cooled in a cooler [b]. In the high pressure pump, liquid carbon
dioxide is pressurized and then heated to a desire temperature
through the heat exchanger [d] to attain supercritical conditions.
The supercritical CO2, then enters to the extraction cell [e] where
it gets loaded at a selected supercritical temperature and pressure.
The seed particles are put into a stainless steel basket placed inside
the extractor for easy and fast charging and discharging of the
extraction cell. The extract rich supercritical-CO2 at the exit of
the extractor flows through a depressurization valve [f]. Conse-
quently, due to the lower pressure, the extracts are released from
the gaseous medium and collected at sample point [g].
2.3. Design of experiments for extraction
The large number of parameters affects the extraction yield
directly or indirectly such as; the extraction temperature, extrac-
tion pressure, particle size, extraction time, amount of co-solvent
(modifier), supercritical CO2 flow rate, bed void fraction, initial
moisture content [11–20]. However, the traditional one factor –
each time method of experimentation, is a cumbersome and time
consuming task, especially when large numbers of variables are
involved in an experiment and also neglect the interaction of these
complex parameters. In this work, extraction temperature, extrac-
tion pressure, seed particle size, supercritical CO2 flow rate and
amount of modifier (%) are used to examine the effects on extrac-
tion yield and other remaining parameters have been kept at favor-
able fixed values such as extraction time at 250 min, to achieve
higher extract, and the amount of seed as 50 g.
The values and levels of five inputs/experimental parameters
for supercritical extraction are reported in Table 2 with an observa-
tion that high pressure is required for vegetable oil extraction
[11,21]. It shows the minimal (Xi, min), the mid range (Xi, mid)
and the maximal (Xi, max) values used for each parameter, which
corresponds to 1, 0 and +1 levels, respectively, in terms of coded
variable Xi defined by Eq. (1).
 
2 X actual  X
Xi ¼ ð1Þ
X i;max  X i;min
X i;max þX i;min
where X ¼ 2
.
The response surface method (RSM) has been used to generate
relationships between above five selected input parameters with
the output parameter as CEY (g oil/g kernel). The design and anal-
ysis of the RSM have been carried out using Design Expert 7.0.3
Software (DE7).
The central composite design (CCD) of RSM, suggested by Box
and Wilson [22], is used to investigate the effect of interactions
of input parameters on the CEY, reduced the extraction time and
cost. It is used to estimate a statistical model that has many differ-
ent combinations of input parameters, as terms in the models and
has at least three levels for each variable. CCD is the most popular
of many classes of RSM designs as it can run sequentially, very effi-
cient and flexible. Among the different classes of CCDs such as
rotatable, spherical, and face centered, face centered design
(a = 1) is simpler one as it requires operating the process at only
356 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365

(e)
Extraction
cell (f)
Metering
valve
(b) (c)
Cooler Pump jacket flow
(a) (g)
CO 2 Sample
Cylinder collection

(d)
Preheater
coil

Fig. 1. Operating method for SFE.

Table 2
Input and output parameters involved in supercritical extraction of vegetable oils.

Factor Name Units Low level Xi, min = 1 Midlevel Xi, mid = 0 High level Xi, max = +1
X1 Temperature °C 60 80 100
X2 Pressure bar 20 30 40
X3 Particle size mm 5 10 15
X4 CO2 flow rate gm/min 0.5 0.75 1.00
X5 Co-solvent % of CO2 flow rate 0 5 10

three levels of each variable in contrast to the other five level
designs. The increased number of reconfigurations in rotatable or
spherical design provides a much greater opportunity for sources
of experimental error, associated with setup and operation, to be
estimated. However, the benefits of rotatable or spherical designs
do not offset the added complexity and associated risk in most
cases. Therefore, the face-centered CCD is found more than ade-
quate for most experiments [23].
The sets of operating parameter as suggested by the DE7 soft-
ware are shown in Table 3. A small face centered central composite
design (SFCCCD) gives 26 different combinations of five different
parameters and consists of 11 factorial points, 10 axial points
and 5 center points. The combination of operating parameters cor-
responding to the central point of the design has been replicated
five times in order to confirm the validity of the model and to
reduce the estimate variance of the values predicted by the model
[24].
Based on the experimental results presented in Table 3, the fol-
lowing second order input–output relationships, called regression
models, for CEY are fitted.
X
5 X
5 X
k1 X
k
Y ¼ b0 þ bi X i þ bii X 2i þ bij X i X j
i¼1 i¼1 i¼1 j>i
where Y is the predicted response, b0 is the coefficient of intercept, bi
is the coefficient of linearity, bii are the quadratic coefficient, bij are
interaction coefficients and Xi, Xj are regressor variables (pressure,
temperature, particle size, CO2 flow rate and co-solvent percentage).
Statistical testing of the models was performed with F-test to
obtain the mathematical relationship between input and output
parameters. To examine the goodness of fit of the model, a test
for significance of regression model was performed and ANOVA
is applied to the response data and the value p < 0.5 is considered
as significant.
2.4. Soxhlet extraction
Soxhlet extraction is the most common method for extraction of
organic compounds from solid samples. Approximately 50 ± 0.2 g
of dried material of watermelon seed is loaded in the glass thimble
separately, which is connected to a 500 ml round bottom flask. The
extraction is carried out using 300 ml n-hexane for 24 h. After
extraction, hexane is removed under reduced pressure by rotary
vacuum evaporator at 50 °C. Experiments are replicated three
times and the oil samples are stored in dark for further analysis.
The amount of extracted oil is then determined gravimetrically.
2.5. Moisture and ash content of watermelon seeds
The moisture content of seed material is determined by stan-
dard method 926.12 recommended by Association of analytical
communities [25]. Samples of quantity measuring 5 ± 0.2 g are
taken into the dried dish and placed in the oven at 105 ± 1 °C. After
1 h dish is removed from the oven and lid is closed. The dish is left
to cool in the desiccators and sample is weighed. The above process
is repeated until a constant weight is achieved for the same
sample. The ash content is determined according to the method
ð2Þ
ISO-749 [26]. For this purpose, 2 ± 0.1 g oil and moisture free seed
sample are taken and heated in an electric furnace at 550 ± 10 °C
until a constant weight is achieved. The percentage of ash content
is calculated in a similar manner as moisture content using Eq. (4).
2.6. Gas-chromatographic analysis of watermelon oil
The high boiling point free fatty acids (FFAs) are converted into
low boiling point fatty acid methyl esters (FAME) by the method
suggested by Christie [27]. The 50 ± 0.001 mg oil sample is taken
in a test tube and dissolved into 1 ml toluene. The test tube is then
fitted with a condenser and 1% sulfuric acid in methanol (2 ml) is
 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365 357

Table 3
Cumulative extraction yield (gm oil/gm kernel) obtained at different experimental conditions proposed by DOE for watermelon oil seeds.

No. X1: X2: Pressure X3: Particle size X4: Solvent flow X4: Co-solvent (% of Experimental yield (gm oil/ Predicted yield (gm oil/
Temperature (bar) (mm) (gm/min) CO2 flow) gm kernel) gm kernel)
(°C)
1 80 300 0.75 10.00 5.00 0.4182 0.4017
2 80 300 0.75 10.00 5.00 0.3951 0.4017
3 80 300 0.75 10.00 5.00 0.4153 0.4017
4 80 300 0.75 10.00 5.00 0.4171 0.4017
5 80 300 0.75 10.00 5.00 0.3816 0.4017
6 60 300 0.75 10.00 5.00 0.3650 0.3701
7 80 300 0.50 10.00 5.00 0.4396 0.4469
8 80 400 0.75 10.00 5.00 0.5181 0.5237
9 80 300 0.75 15.00 5.00 0.5066 0.5050
10 80 300 0.75 10.00 0.00 0.2643 0.2701
11 80 300 0.75 10.00 10.00 0.4724 0.4802
12 80 200 0.75 10.00 5.00 0.0818 0.0898
13 80 300 1.00 10.00 5.00 0.2945 0.3007
14 80 300 0.75 5.00 5.00 0.2977 0.2985
15 100 300 0.75 10.00 5.00 0.4461 0.4545
16 100 400 0.50 15.00 0.00 0.5183 0.5193
17 100 400 0.50 5.00 10.00 0.4682 0.4677
18 100 200 0.50 15.00 10.00 0.2641 0.2636
19 60 200 1.00 15.00 10.00 0.2382 0.2392
20 100 400 1.00 5.00 0.00 0.1764 0.1789
21 100 200 1.00 15.00 0.00 0.0471 0.0496
22 60 400 1.00 15.00 0.00 0.3200 0.3213
23 60 400 0.50 15.00 10.00 0.4786 0.4787
24 100 200 1.00 5.00 10.0 0.0522 0.0532
25 60 200 0.50 5.00 0.00 0.0684 0.0672
26 60 400 1.00 5.00 10.00 0.4474 0.4484

added to the test tube. The mixture is heated at 70 °C with total
reflux conditions. After two hours, freshly prepared 5% sodium
chloride solution is added and converted FAME is extracted two
times with 5 ml n-hexane. The upper n-hexane layer is separated
with the help of Pasteur pipette. Then the n-hexane layer is washed
with 4 ml freshly prepared 2% potassium bicarbonate solution and
dried over the anhydrous sodium sulfate. The solvent is removed
under reduced pressure by a rotary vacuum evaporator.
The FAME is analyzed using Thermo Trace Ultra Gas chromato-
graph (Thermo Scientific, USA) equipped with HP 88 capillary col-
umn of 30 m length, 0.25 mm diameter and 0.20 lm film
thickness (Agilent Technologies, USA) with a flame ionization detec-
tor (FID). The nitrogen gas with a flow rate of 30 ml/min is used as a
makeup gas. The H2 and air flow rate are maintained at 35 ml/min,
350 ml/min respectively. The injector (FID) temperature is main-
tained at 250 °C. A temperature program with a total run time of
44.5 min is used. The initial column temperature is maintained at
120 °C for 1 min. Then oven temperature is increased to 145 °C at
a rate of 5 °C/min and kept constant at this temperature for 1 min.
The temperature again increased from 145 °C to 220 °C at a ramp
rate of 2 °C/min and kept constant at 220 °C for 2 min. The prepared
FAME is makeup with 1 ml n-hexane and 1 lL is injected by an auto-
sampler (HP 7683, HP Company, Wilmington, DE). The split ratio is
kept as 50:1. The peak areas and data collection are managed using
a chrom-cad (Revision. A.09.01, Thermo Scientific, USA) software.
Fatty acid peaks are identified using a standard 37 FAME mixture
(Supelco 37 component FAME mix, Supelco, Bellefonte, PA).
3. Results and discussion
The salient results of the present investigation are discussed
below:
3.1. Total oil yield, moisture and ash content
The total oil content in seed material has been investigated by
Soxhlet extraction and is found approximately to be 52.37%. The
value of total oil extracted with n-hexane is comparable with the
reported data of Acar et al. [3] who obtained 52.34% oil yield with
the watermelon seed obtained from the local market of Konya, Tur-
key. However, this value is much higher as compared to the 32.17%
yield reported by Conto et al. [7]. Such variation may be attributed
to variation in geographical condition such as soil properties, nutri-
ents, water availability and seasonal temperature [8]. The moisture
and ash content of watermelon seeds are found to be approxi-
mately 6.93% and 4.64% respectively.
3.2. Thermal characterization
Thermal degradation of the watermelon seed kernel is investi-
gated by thermo gravimetric analysis (TGA) as represented in
Fig. 2. The plots can be divided into three parts. The first part of
the thermogram, from 22 °C to 130 °C, is associated with dehydra-
tion of seed kernels and a weight loss of 4.177% (wt%) is observed.
The second part of the thermogram, which is from 130 °C to 500 °C,
is associated with the decomposition of organic matter present in
the seed kernels and a weight loss of 80.95% (wt%) is observed.
Similarly, the third part, from 500 °C to 1015 °C, is related to
decomposition of most of the component of seed kernel and a
weight loss of 4.59% (wt%) is observed. At a temperature of
1015 °C there is around 10.28% of residue left, due to the presence
of ash which probably contains inorganic oxides.
3.3. FTIR analysis
Fourier transform infrared (FTIR) spectroscopy is an important
technique for the qualitative analysis of organic compounds, pres-
ent in the natural products. The FTIR analysis of watermelon seed
kernel has been done in the dried state to identify the various func-
tional groups and also indicate the complex nature of this material.
The FTIR spectrum of dried and power watermelon seed is mea-
sured, as KBr pellet, in the range 400–4000 cm1 and is shown in
Fig. 3. FTIR spectra of watermelon seed show various peaks; how-
ever, only sharp peaks are considered.
The bandwidth centered at wavelength 3309 cm1 may be
attributed to the stretching of O–H bonds present in fatty acids,
358 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365

120 seed particles. The seed samples, whether non-extracted or

extracted, are first coated with gold–palladium alloy. The SEM
has been performed using LEO-1550 scanning electron microscope.
100
Fig. 4(a) and (b) shows the SEM images of watermelon seed of
0.75 mm particle diameter. The structure examination of all seed
80 material, non-extracted (a) and extracted (b) is carried out from
the SEM images at 500 magnifications. The images of non-
TG (wt%)

extracted seeds exhibit an uneven and rough surface. It appeared

60
40
20
0
0 200 400
Temperature ( C)
Fig. 2. TG thermograms for watermelon eeed.
carbohydrates, and proteins [28–30]. The peaks observed at
2924 cm1 and 2856 cm1, respectively, correspond to asym-
metric and symmetric stretching of the C–H bond of the CH2 group
present in fatty acids [31–33]. Watermelon seeds also have a high
content of protein and hence it also contributes toward N–H
stretching of the amide group. The carbonyl component that
appears due to the presence of lipids or ester can be seen at
1743 cm1 [30,31], and the peak that appears at 1632 cm1 is
attributed to the vibrational frequency of carbonyl group of amides
present in the protein portion [30]. The peak that appears at
1542 cm1 supports the carbonyl functional group present in
the amide II region shows the presence of a-helices structure of
the protein or shows the deformation of the amide N–H stretch
[33]. The –C–H bending of alkanes observed at the absorption
wavelength 1456 cm1 (1466–1437 cm1) and the peak at wave-
length 1234 cm1 (1246–1016 cm1) support the stretching
vibration of the (–C–O) ester groups [31]. The peak at
1153 cm1 shows the NH3 rocking.
3.4. Study of structure of samples using scanning electron microscopy
(SEM)
Scanning electron microscopy (SEM) analysis technique has
been used to observe the surface morphology of the watermelon
from the microphotographs of all non-extracted seed particles that
the particle surfaces of seed are formed by a sequence of layers and
oil and are uniformly distributed in every layer. SEM images clearly
show that oil and non-extracted solid phase are closely interpene-
trating. However, for the extracted seed the layers are found
cracked and pour clearly appeared as shown in (b) part of the
image (Fig. 4). This indicates that high pressure is required to rup-
ture the layer of the seed material for better oil extraction.
600 800 1000
o
3.5. Response surface analysis
Experimental results obtained for CEY, reported in Table 3, are
analyzed using ANOVA to figure out the best correlation between
CEY and input parameters. The three available models such as lin-
ear, two-factor interaction model (2FI) and quadratic are tested for
ANOVA and the results of ANOVA analysis are discussed below.
3.5.1. Linear model
First of all, the regressed linear model as given by Eq. (4) is fitted
considering all the five input parameters.
Yield ðg oil=g kernelÞ ¼ 0:0856  2:6797  104  X 1
þ 1:4610  103  X 2  0:2261  X 3
þ 0:0105  X 4 þ 0:0128  X 5 ð3Þ
An ANOVA is performed to establish the relative significance of
the individual factors and found that X2, X3 and X5 are the signifi-
cant parameters, whereas X1, X4 are insignificant. The p-value for
the model is less than 0.0001 but correlation coefficients, ‘‘R-
square’’, ‘‘adjusted R-square’’ and ‘‘predicted R-square’’ values for
Eq. (4) are 0.7079, 0.6349 and 0.3860, respectively. Thus, it can
be said that Eq. (4) offers low ‘‘predicted R-square’’ and very small
‘‘R-square’’ values. The model can be improved by removing the
insignificant model terms. But to maintain the hierarchy, these
terms were kept in the model according to the hierarchy principle
[23].

100

95

90

85

80
%Transmittance

75
432.16

70

65
1632.49

1453.99
3482.70

448.18
721.46
2855.18

1743.73

1153.20

60
2924.84

55

50

45

4000 3500 3000 2500 2000 1500 1000 500

Wavenumbers (cm-1)

Fig. 3. FT-IR spectra of watermelon oil seeds.

 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365 359

3.5.2. Two-factor interaction model
The regressed 2FI model is given by Eq. (4). An ANOVA analysis
is performed and it is found that only one individual parameter X2
is significant and all other parameter terms, interaction terms are
insignificant.
Yield ðg oil=g kernelÞ ¼ 1:1184  7:458  103  X 1
model, Eq. (5), can be improved by removing the insignificant
model terms. The removal of insignificant terms one by one in a
stepwise manner (starting from largest insignificant to smallest
insignificant term) was carried out to improve the model which
produced higher ‘‘R-square’’ or ‘‘predicted R-square’’. The regressed
reduced quadratic model is given by Eq. (6). The regression coeffi-
cients of regression analysis are given in Table 5

þ 2:0763  103  X 2  2:8435  X 3 Yield ðg oil=g kernelÞ ¼ 0:1033  8:9073  103  X 1

 0:0178  X 4  0:0399  X 5
6
 5:3994  10  X 1 X 2 þ 0:0116  X 1 X 3
þ 1:955  104  X 1 X 4 þ 8:7012  105  X 1 X 5
3 5
þ 1:901  10  X 2 X 3  7:2398  10
5
 3:2948  10  X 2 X 5 þ 0:0613  X 3 X 4
4
þ 0:0881  X 3 X 5  2:4395  10  X4X5
The correlation coefficients, ‘‘R-square’’, ‘‘adjusted R-square’’
and ‘‘predicted R-square’’ values for Eq. (4) are 0.8117, 0.5292
and 36.2282 respectively. As Eq. (4), offers negative ‘‘predicted
R-square’’ value, the model can be improved by removing the insig-
nificant model terms. The ‘‘Adequate precision’’ value of 5.838
indicates an adequate signal. For the present model, the lack of
fit is significant. Thus, taking all the above facts into consideration,
it can be concluded that the 2FI model is not an effective model.
3.5.3. Development of quadratic model
The quadratic model is developed using the experimental CEY
values reported in Table 3. The regressed quadratic model is given
by Eq. (5).
þ 8:2065  103  X 2  2:0018  X 3
 8:5360  103  X 4  0:0129  X 5
 8:2759  106  X 1 X 2 þ 0:0105  X 1 X 3
 X2X4 þ 1:3798  104  X 1 X 4
þ 1:7771  103  X 2 X 3
ð4Þ  7:8597  105  X 2 X 4
 4:4454  105  X 2 X 5 þ 0:0588  X 3 X 4
þ 0:0835  X 3 X 5  4:7407  104  X 4 X 5
þ 2:6528  105  X 21  9:4989  106  X 22
 0:4462  X 23  1:0636  103  X 25 ð6Þ
ANOVA results for reduced quadratic model for CEY are given in
Table 4. From the table, it is perceptible that the individual parame-
ter terms and second order interaction terms such as X1, X2, X3, X4, X5,
X1X2, X1X3, X1X4, X2X3, X2X4, X2X5, X3X4, X3X5, X4X5, X21, X22, X23 and X25 are
significant. The p-value of the reduced quadratic model is less than
0.001 which shows excellent fit of experimental data. ‘‘Adequate
precision’’ of 38.454 indicates an adequate signal and the ‘‘lack of

Yield ðg oil=g kernelÞ ¼ 0:12936  8:8936  103  X 1

þ 8:3620  103  X 2  1:93960  X 3
 0:01133  X 4  0:0166  X 5
 7:9371  106  X 1 X 2 þ 0:0106  X 1 X 3
þ 1:4476  104  X 1 X 4
þ 3:6258  105  X 1 X 5
þ 1:6980  103  X 2 X 3
 8:2548  105  X 2 X 4
 4:3098  105  X 2 X 5 þ 0:0572  X 3 X 4
þ 0:0840  X 3 X 5  4:4697  104  X 4 X 5
þ 2:3247  105  X 21  9:6301  106  X 22
 0:4672  X 23 þ 2:3595  104  X 24
 1:1161  103  X 25 ð5Þ
ANOVA results for the quadratic model for CEY suggested that
the individual parameters and second order interaction terms such
as X1, X2, X3, X4, X5, X1X3, X2X3, X2X4, X3X4, X3X5, X22X23 and X25 are sig-
nificant, whereas all other terms are insignificant. The correlation
coefficients, ‘‘R-square’’, ‘‘adjusted R-square’’ and ‘‘predicted R-
square’’ values for Eq. (5) are 0.9976, 0.9879 and –3.5506 respec-
tively. Eq. (5) offers negative ‘‘predicted R-square’’, which implies
that the overall mean is a better predictor of the response than
the current model. However, adequate precision of 32.03 indicates
an adequate signal and the lack of fit for the present model is
insignificant.
Thus, taking all the above facts into consideration, it can be con-
cluded that the quadratic model is not a good model. Further, for
the model, the ‘‘R-square’’ value is higher, but the model contains
a number of insignificant terms and thus leading to an inferior Fig. 4. (a) Surface of watermelon seed before extraction and (b) surface of
model and hence negative ‘‘predicted R-square’’ value. Thus, the watermelon seed after extraction.
360 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365

Table 4
ANOVA for RSM variables fitted to reduced quadratic model, Eq. (6).

Source Sum of squares df Mean square F-value p-value Significance

Model 0.56 18 0.031 148.90 <0.0001 Significant
X1: Temperature 4.164  103 1 4.164  103 19.91 0.0029 Significant
X2: Pressure 0.11 1 0.11 538.15 <0.0001 Significant
X3: Particle size 0.013 1 0.013 62.14 0.0001 Significant
X4: CO2 flow rate 0.026 1 0.026 121.91 <0.0001 Significant
X5: Co-solvent% 0.026 1 0.026 126.58 <0.0001 Significant
X1X2 3.709  104 1 3.709  104 1.77 0.2247 Insignificant
X1X3 3.710  103 1 3.710  103 17.74 0.0040 Significant
X1X4 2.578  104 1 2.578  104 1.23 0.3036 Insignificant
X2X3 3.731  103 1 3.731  103 17.84 0.0039 Significant
X2X4 2.920  103 1 2.920  103 13.96 0.0073 Significant
X2X5 6.688  104 1 6.688  104 3.20 0.1169 Insignificant
X3X4 0.010 1 0.010 48.83 0.0002 Significant
X3X5 0.015 1 0.015 70.50 <0.0001 Significant
X4X5 1.902  104 1 1.902  104 0.91 0.3721 Insignificant
X21 2.902  104 1 2.902  104 1.39 0.2773 Insignificant
X22 0.023 1 0.023 111.18 <0.0001 Significant
X23 2.005  103 1 2.005  103 9.58 0.0174 Significant
X25 1.822  103 1 1.822  103 8.71 0.0214 Significant
Residual 1.464  103 7 2.092  104
Lack of fit 3.931  104 3 1.310  104 0.49 0.7083 Insignificant
Pure error 1.071  103 4 2.678  104
Cor total 0.56 25

Table 5
0.6
Regression analysis for reduced quadratic model, Eq. (6).
Pridicted Extraction Yield (gm oil/gm kernel)

Predictor Coefficient
0.5
Actual Coded
Constant 0.10329 0.40 0.4
9.73%
X1: Temperature 8.90731  103 0.042
X2: Pressure 8.20654  103 0.22 -3.94%
X3: Particle size 2.00178 0.074 0.3
X4: CO2 flow rate 8.53604  103 0.10
X5: Co-solvent% 0.012866 0.11
X1X2 8.27589  106 0.017 0.2
X1X3 0.010470 0.052
X1X4 1.37982  104 0.014
0.1
X2X3 1.77707  103 0.044
X2X4 7.85967  105 0.039
X2X5 4.44535  105 0.022 0.0
X3X4 0.058801 0.074 0.0 0.1 0.2 0.3 0.4 0.5 0.6
X3X5 0.083499 0.10 Experimental Extraction Yield (gm oil/gm kernel)
X4X5 4.74071  104 0.012
X21 2.65280  105 0.011
Fig. 5. Parity plot for the reduced quadratic model, Eq. (6), developed for prediction
X22 9.49888  106 0.095
of extraction yield (gm oil/gm kernel).
X23 0.44622 0.028
X25 1.06355  103 0.027

fit’’ is insignificant (p > 0.05). The p-value of the ‘‘lack of fit’’ is 0.7083,
which suggests that insignificant lack of fit is good for data fitness to
the model. However, the correlation coefficients, ‘‘R-square’’,
‘‘adjusted R-square’’ and ‘‘predicted R-square’’ values for Eq. (6)
are 0.9974, 0.9907 and 0.9226, respectively. The ‘‘Predicted R-
Square’’ of 0.9226 is in good agreement with the ‘‘Adjusted R-
Square’’ of 0.9907.
Thus, it can be concluded that Eq. (6) offers 92.26% (predicted R-
Square) of the variability in predicting new observations in com-
parison with approximately 99.74% (R-square) variability in the
original data. Thus, this model can be used to navigate the design
space. Now, taking all the facts into consideration, it can be con-
cluded that the reduced quadratic model is the best model.
The parity plot for the reduced quadratic model, given by Eq.
(6), for the prediction of CEY, is shown in Fig. 5. The error band
extends from –3.94% to +9.73%, and all data points fall within this
error band. Thus, the CEY predicted by Eq. (8) lies within 3.94% to
+9.73% of experimental values.
The error% is computed using the formula
YðPÞ  YðEÞ
Errorð%Þ ¼  100 ð7Þ
YðEÞ
where Y(P) is the value of the predicted output obtained using the
model equation, Eq. (6), and Y(E) is the experimental output
obtained from experiments.
Based on the above discussed criteria for individual models, it
can be concluded that the reduced quadratic model described by
Eq. (6) for the CEY, fits the experimental values best in comparison
with other two models, i.e. linear and 2-factor interaction models.
This is because of the fact that the error band for this model is
small and the ‘‘R-square’’, ‘‘adjusted R-Square’’ and ‘‘predicted R-
Square’’ values are close to each other and also the error band con-
tains all the data points.
3.6. Effect of various parameters on CEY
The effect of various input parameters, such as temperature
(X1), pressure (X2), particle size (X3), CO2 flow rate (X4) and co-
solvent% (X5), on the CEY (g oil/g kernel) of watermelon seed oil is
 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365 361

Perturbation Fig. 6 shows that CEY is highly sensitive to pressure as compared to

0.53 B other parameters and temperature has the lower effect on the CEY.
Yield D
E The other parameters such as, particle size, CO2 flow rate and co-
Actual Factors C A solvent percentage have a moderate effect on CEY. The extraction
A: Tem (X1) = 80.00
B: press (X2) = 300.00 0.4075 temperature, pressure, CO2 flow rate and co-solvent percentage
C: PS (X3) = 0.75
D: flow (X4) = 10.00
A have a positive effect, whereas particle size has a negative effect
E: Co (X5) = 5.00 on the CEY.
Yield D C
0.285 E
3.6.2. Effect of 2-parameter interactions on CEY (g oil/g kernel)
To observe the effect of interaction of various input parameters,
0.1625 such as temperature (X1), pressure (X2), particle size (X3), CO2 flow
rate (X4) and co-solvent% (X5), on the CYE (g oil/g kernel) of water-
B
melon seed oils, two factor interaction plots have been plotted as
0.04 given in Fig. 7(a)–(i). The 2-parameter interaction response surface
plot obtained from DE7 has been plotted for the different input
-1.000 -0.500 0.000 0.500 1.000 parameters controlling CEY while other parameters are constant
Deviation from Reference Point (Coded Units) at the base case as shown in Table 6. It is found that, all the input
parameters were found to be involved in interactions within the
Fig. 6. Effect of individual parameters on watermelon cumulative extraction yield
range of parameters studied in the present investigation.
(gm oil/gm kernel).
Fig. 7(a), (d), (e) and (f) shows the simultaneous effects of
pressure and temperature, pressure and particle size, pressure
and CO2 flow rate & pressure and the addition of co-solvent (%)
Table 6
on the CEY at the flow rate of 10 g/min respectively. It can be
Base case for determining the effects of parameters in MPC.
seen that dependency of CEY of oil is large on pressure as com-
Name of input parameters Base case Range pared to temperature. At a given temperature or particle size or
Lower Upper flow rate or co-solvent (%), CEY increases with increasing pres-
X1: Temperature 80 60 100 sure and can be explained by the fact that, as pressure increases
X2: Pressure 300 200 400 solvent density increases hence, the interaction between the oil
X3: Particle size 0.75 0.50 1.00 and CO2 molecules increases, which lead to the increase of oil
X4: CO2 flow rate 210 180 240 solubility in CO2.
X5: CO-solvent% 05 00 10
Similarly, sometime it is very difficult to isolate the effect of
temperature than pressure as both have similar effects on CEY.
The increase in CEY with the increase in temperature can in
found by perturbation plot. A perturbation plot is like one factor- one hand be due to the decrease in supercritical carbon dioxide
at-a-time experimentation and does not show the effect of density and, on the other hand, due to the enhanced vapor pres-
parameter interactions. It is very helpful to compare the effect of sure of fatty acid ingredients [19]. As shown in Fig. 7(a), at low
all independent variables at a particular point in the design space. pressure and/or larger particle size, temperature has larger
The response is plotted by changing only one factor over its range impact on the CEY as compared to high pressure and increasing
while holding the other factors constant at the base case as shown the temperature from 60 to 100 °C favors the extraction because
in Table 6. A steep slope or curvature in a factor shows that the of increase of vapor pressure of fatty acids which dominates over
response is sensitive to that factor and relatively flat line shows the decrease in CO2 density effect. Fig. 7(b) shows the interaction
insensitivity to change in that particular factor [34]. of particle size and temperature, Fig. 7(g) shows the interaction of
particle size and addition of co-solvent (%) to CO2 rate and simi-
3.6.1. Effect of individual parameter on CEY larly Fig. 7(h) shows the interaction of particle size and CO2 flow
To understand the effect of each parameter on CEY Fig. 6 is plot- rate (appears to be repeated see blue marking). At lower temper-
ted. The mechanism of the supercritical fluid extraction process ature and/or lower co-solvent percentage and/or lower CO2 flow
can be explained by the following steps: (1) Transport of supercrit- rate, size of particle plays an important role to increase the
ical solvent to the particle surface and then from the particle sur- CEY. As the particle size increases from 0.500 mm to 1.000 mm,
face to interior of particle by diffusion; (2) Dissolution of the the extraction yield decreases considerably. At higher tempera-
solute with the supercritical solvent; (3) Transport of supercritical ture and/or higher co-solvent percentage and/or higher CO2 flow
solvent with molecules from interior of particle to particle surface; rate, the effect of particle size on CEY is not much. The effect of
and (4) Transport of supercritical solvent and solute molecules particle size on CEY can be explained in terms of mass transfer
from the particle surface to bulk solvent [16]. resistance. The extraction of solute from the solid matrix offers
The behavior of CEY plot is explained by effect of each parame- two types of mass transfer resistance, namely internal mass
ter on CEY in terms of mass transfer coefficients coupled with transfer resistance and external mass transfer resistance. If the
effective intra-particle diffusion coefficient in steps 1–5. It is noted internal mass transfer mechanisms constitute the controlling step
that for a given time period, the CEY increases with increase in of the extraction process, the particle size of the vegetable matter
temperature. The observation can be explained by the fact that can significantly influence the CEY. In this case, extraction from
the mass transfer coefficients increase with rise in temperature different particle sizes will largely depend on the length of the
as a result of high diffusivity of watermelon seed oil in SC-CO2 at diffusion path. If external mass transfer or equilibrium is the con-
higher temperatures. Thus the rate of mass transfer of solute to trolling steps of the process, the particle size cannot affect the
bulk liquid phase increases with increase in temperature. Similarly, extraction rate very much [20]. It is observed from Fig. 7(c), (e),
CEY increases with increase in pressure. The observation can be (h) and (i), that the CEY increases significantly with the increase
explained by the fact that the solubility of watermelon seed oil in CO2 flow rate from 5 g/min to 15 g/min. It can be safely con-
in SC-CO2 increases at higher pressures despite the fact that mass cluded that, CEY increases with the increase of the flow rate
transfer coefficients decrease with a rise in pressure. Apart from it, because as the flow rate increases, thickness of the film layer
362 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365

0.57 0.47

0.4375 0.405

0.305 0.34
Yield Yield
0.1725 0.275

0.04 0.21

400.00 100.00 1.00 100.00

350.00 90.00 0.88 90.00
300.00 80.00 0.75 80.00
B: press (X2) 250.00 70.00 A: Tem (X1) 0.63 70.00
C: PS (X3) A: Tem (X1)
200.00 60.00 0.50 60.00

(a) (b)

0.58 0.53

0.5025 0.3825

0.425 0.235
Yield Yield

0.3475 0.0875

0.27 0.06
-

15.00 100.00 1.00 400.00

12.50 90.00 0.88 350.00
10.00 80.00 0.75 300.00
7.50 70.00 0.63 250.00
D: flow (X4) A: Tem (X1) C: PS (X3) B: press (X2)
5.00 60.00 0.50 200.00

(c) (d)

0.59 0.58

0.4275 0.415

0.265 0.25
Yield
Yield
0.1025 0.085

0.06
-
0.08
-

15.00 10.00 400.00

400.00
7.50 350.00
12.50 350.00
5.00 300.00
10.00 300.00
2.50 250.00
7.50 250.00 B: press (X2) E: Co (X5) B: press (X2)
D: flow (X4) 0.00 200.00
5.00 200.00

(e) (f)

Fig. 7. Effect of two parameter interaction on watermelon cumulative extraction yield.

around the solid particles reduces and thus mass transfer resis- concluded that CEY increases substantially with the increase in
tance becomes small. As a result, CEY increases [14,35,36]. co-solvent (%). Fig. 7(g) shows that, the addition of co-solvent with
The effect of ethanol as a polar co-solvent (%) on the CEY can be larger particle has a larger impact on CEY. The addition of a co-
observed from Fig. 7(f), (g), and (i). From the figures it can be solvent to a supercritical fluid, generally, increases the polarity,
 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365 363

0.49 0.51

0.3825 0.4125

0.275 0.315
Yield Yield
0.1675 0.2175

0.06 0.12

10.00 1.00 15.00 1.00

7.50 0.88 12.50 0.88
5.00 0.75 10.00 0.75
E: Co (X5) 2.50 0.63 C: PS (X3) 7.50 0.63
D: flow (X4) C: PS (X3)
0.00 0.50 5.00 0.50

(g) (h)

0.58

0.4725

0.365
Yield
0.2575

0.15

10.00 15.00
7.50 12.50
5.00 10.00
E: Co (X5) 2.50 7.50 D: flow (X4)
0.00 5.00

(i)
Fig. 7 (continued)

solute–solvent interaction and bulk density of the fluid mixture
which would contribute to solubility enhancement. The hydroxyl
group present in ethanol may lead to form hydrogen bond to
increase the solubility of oils [37]. Since, the solubility enhance-
ment with the addition of a co-solvent is mainly caused by the for-
mation of special interactions between the solute and co-solvent
molecules, solubility of oils in supercritical carbon dioxide greatly
enhanced with the addition of ethanol as a co-solvent. Cocero et al.
[38] successfully demonstrated that the solubility of sunflower oil
increases with increasing in ethanol percentage as a co-solvent and
also suggested that, the extraction of some phospholipids is
directly proportional to the added ethanol volume which explains
the higher CEY of watermelon seed oil.
3.7. Fatty acid (unsaturated and saturated)
The CEY for SC-CO2 extraction is somewhat lower than that of
hexane extraction. As both solvent, hexane and SC-CO2, are non-
polar they should exhibit similar behavior in the extraction pro-
cesses. The observed difference in the oil yield between these
two methods could be due to the high pressure and different
extraction times. As said earlier, the extraction time for the Soxhlet
method is six times longer than that of Supercritical fluid
extraction.
The fatty acid compositions (saturated and unsaturated) of the
oil extracted by supercritical CO2 and by hexane are given in
Table 7 and GC chromatographs are given in Fig. 8. Table 7 shows
that, oil comprised of unsaturated fatty acid ranging from 73.597%
to 75.597% and saturated fatty acids ranging from 26.634% to
24.182%. The average value of saturated and unsaturated fatty
acids present in WM oil are 25.12% and 74.23% respectively. Table 7
shows that linoleic acid (C18:2 n6c) is a major unsaturated fatty
acid (61%) in watermelon seed oil, which is in close agreement
with those reported by Milovanovic and Picuric-Jovanovic [6]
(62.2%), and a little bit higher as those reported by El-Adawy and
Taha [39] (59.64%) and lower than those reported by Acar et al.
[3] (63.19%), Conto et al. [7] (65.61%), Baboli and Kardi [40]
(68.3%). Watermelon seed oil is a high source of linoleic acid
(C18:2 n6c), is important polyunsaturated omega-6 fatty acid,
helps to skin, bone and hair growth, controls metabolism and
maintains the reproductive system. Omega-6 also plays an impor-
tant role in the nervous system, brain and muscle growth [41]. The
watermelon seed oil is also rich in cis-oleic acid (C18:1 n9c)
(11.275–14.325%), which is an important monounsaturated
omega-9 fatty acid because of its stability and health benefit prop-
erties such as treatment and prevention of heart disease, decreases
the chances of cardiovascular disease [42,43].
From Table 7, it is noted that, small amounts of palmitoleic acid
(C16:1) (0.04–0.091%) and eicosenoic acid (C20:1) (0.019–0.346%)
also are adding up to unsaturation of watermelon oil. In the entire
oil composition, saturated fatty acids such as, myristic acid (C14:0),
palmitic (C16:0), Stearic (C18:0) and arachidic (C20:0), contribute
364 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365

Table 7
Fatty acid composition of watermelon seed oil.

Run Compound
C14:0 C16:0 C16:1 C18:0 C18:1 n9c C18:2 n6c C20:0 C20:1 SFA USFA
1 0.058 13.602 0.062 11.959 12.737 60.885 0.554 0.142 26.173 73.827
2 0.095 12.882 0.053 11.317 13.025 62.015 0.482 0.130 24.776 75.223
3 0.087 12.561 0.049 11.403 12.860 62.426 0.462 0.152 24.513 75.487
4 0.053 12.530 0.072 11.328 12.851 62.542 0.492 0.132 24.403 75.597
5 0.061 12.835 0.042 11.465 12.642 62.090 0.529 0.333 24.890 75.107
6 0.063 12.647 0.512 10.903 12.615 62.185 0.829 0.246 24.442 75.558
7 0.073 12.769 0.051 10.989 12.660 62.592 0.702 0.164 24.533 75.467
8 0.088 12.698 0.048 11.830 12.598 62.270 0.449 0.019 25.065 74.935
9 0.075 12.259 0.052 11.830 12.667 62.513 0.452 0.152 24.616 75.384
10 0.068 12.376 0.057 11.623 12.690 62.512 0.520 0.154 24.587 75.413
11 0.076 12.797 0.051 11.455 12.522 62.156 0.532 0.384 24.860 75.113
12 0.082 12.212 0.062 11.623 12.623 62.429 0.623 0.346 24.540 75.460
13 0.060 12.595 0.049 10.956 12.273 62.369 1.430 0.268 25.041 74.959
14 0.058 12.726 0.041 13.245 11.275 61.962 0.605 0.088 26.634 73.366
15 0.079 13.160 0.052 11.522 12.294 62.179 0.495 0.219 25.256 74.744
16 0.085 13.428 0.048 11.447 12.516 61.993 0.441 0.042 25.401 74.599
17 0.069 12.462 0.051 11.758 13.140 60.407 1.989 0.124 26.278 73.722
18 0.083 13.883 0.050 10.325 12.604 61.712 0.433 0.910 24.724 75.276
19 0.072 13.442 0.049 11.344 12.949 61.396 0.344 0.404 25.202 74.798
20 0.073 13.523 0.052 11.497 12.324 61.462 0.643 0.426 25.736 74.264
21 0.103 13.521 0.049 09.798 15.002 60.672 0.772 0.083 24.194 75.806
22 0.091 12.872 0.048 11.928 12.195 62.130 0.606 0.120 25.497 74.493
23 0.082 12.392 0.047 11.124 14.325 61.371 0.584 0.075 24.182 75.818
24 0.103 14.588 0.052 11.074 13.172 60.297 0.571 0.143 26.336 73.664
25 0.091 13.827 0.091 10.691 13.221 61.422 0.501 0.156 25.110 74.890
26 0.087 12.957 0.073 11.523 13.726 60.990 0.482 0.162 25.049 74.951
Hexane Extracted 0.084 13.350 0.086 11.738 13.823 60.231 0.562 0.126 25.734 74.266

SFA: Saturated Fatty Acids, USFA: Unsaturated Fatty Acids.

Fig. 8. Gas chromatograph of supercritical CO2 extracted watermelon seed oil.

0.53–0.95%, 12.212–14.588%, 9.798–11.928% and 0.033–1.989%
share respectively in the total oil composition. It is noticeable from
Table 6.64, that the composition of supercritical CO2 and hexane
extracted watermelon oils are almost same and no significant var-
iation in the composition of the fatty acid is observed. Apart from
it, oil extracted by supercritical CO2 is free from organic solvent
such as hexane improving the quality of WM oil.
4. Conclusion
The present work studies the performance of supercritical fluid
extraction of watermelon seeds with the help of small face
centered composite design of response surface methodology by
using five important input parameters such as extraction pressure,
temperature, particle size, CO2 flow rate and modifier percentage.
From the above work, it can easily be concluded that reduced sec-
ond order polynomial model successfully predicts the CEY with the
change in independent parameters within the experimental
ranges. The results indicate that, all the individual parameters have
a significant effect (p < 0.05) on CEY. The watermelon oil is highly
unsaturated with rich in linoleic acid, which makes it suitable for
edible purpose and a high source of protein. It should show the
percent of error of fitting, interaction parameters, CEY of water-
melon seed, etc.
 A. Rai et al. / Separation and Purification Technology 141 (2015) 354–365 365

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