Procedia Chemistry 9 ( 2014 ) 265 272

1876-6196 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of the Organizing Committee of ICCE UNPAR 2013
doi: 10.1016/j.proche.2014.05.032
Available online at www.sciencedirect.com
ScienceDirect
International Conference and Workshop on Chemical Engineering UNPAR 2013, ICCE UNPAR
2013
Application of supercritical fluid extraction on food processing:
black-eyed pea (Vigna unguiculata) and peanut (Arachis hypogaea)
Karmelita Anggrianto, Rinaldi Salea, Bambang Veriansyah, Raymond R Tjandrawinata*
Advanced Technology Development, Dexa Laboratories of Biomolecular Sciences (DLBS), Dexa Medica
Industri Selatan V, Block PP no. 7, Jababeka Industrial Estate II, Cikarang, West Java 17550, Indonesia
Abstract
This present study investigates application of supercritical fluid carbon dioxide extraction (SCFE-CO
2
) for removing fats from
black-eyed pea (Vigna unguiculata) and peanut (Arachis hypogaea) to produce high protein-low fat diet products. SCFE-CO
2
process used Taguchis orthogonal array at three parameters in three levels: pressures between 25-35 MPa, temperatures between
40-60
o
C and CO
2
flowrates between 10-20 g/min. The results of the experiment showed that the optimum conditions of SCFE-
CO
2
process were 25 MPa, 60
o
C, 10 g/min for black-eyed pea and 35 MPa, 60
o
C, 15 g/min for peanut. The highest yield for
black-eyed pea and peanut were 5.4% and 48.5%, respectively.
2014 Anggrianto et al. Published by Elsevier B.V.
Selection and peer-review under responsibility of the Organizing Committee of ICCE UNPAR 2013.
Keywords: Black-eyed pea; peanut; supercritical fluid extraction; taguchi method.
Nomenclature
Adj MS adjusted mean of square
Adj SS adjusted sum of square
DF degree of freedom
Pc critical pressure (MPa)
Q flow rate
Seq SS sequenced sum of square
* Corresponding author. Tel.: +62-21-898-41901; fax: +62-21-898-41905 .
E-mail address: raymond@dexa-medica.com.
2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of the Organizing Committee of ICCE UNPAR 2013
266 Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272
SF solvent/feed ratio
S/N signal-to-noise
Tc critical temperature (C)
1. Introduction
Recently, demand for low calorie and low fat healthy food is increasing for diet nutrition. The trend is promoted
by medical reports that high calorie or high fat diet will increase the risk of metabolic disease namely diabetes and
obesity that lead to cardiovascular disease
1-3
. Low calorie diet generally has high protein or fiber to promote cell
regeneration and digestion tract health. High protein-low calorie diet can be sourced from animal, such as whey
protein, or from plant, such as seed from Fabaceae family or Leguminosae. The Fabaceae family or Leguminosae,
is commonly known as the legume, pea or bean family, are a large and economically important family of flowering
plants. Legume seeds have high protein content and is used for human and animal consumption or for production of
edible oils. Peanut (Arachis hypogaea) from Genus Arachis is a native to central South America. Peanut has become
a significant agricultural commodity and its oil is a primary ingredient in the culinary process in many countries
4
.
Peanut contains approximately 38% oil w/w
5
and 25-28 % w/w of protein
6
. Black-eyed pea (Vigna unguiculata) is
another important grain legumes which forms the source of dietary protein
7,8
. The black-eyed pea protein content is
relatively high between 19.5 % and 27.3 % w/w
9
. In terms of amino acids, black-eyed pea seeds were considered as
rather well balanced in essential amino acids. This seeds are traditionally used for astringent, appetizer, laxative,
anthelmintic, aphrodisiac, diuretic, galactgoue and liver tonic
7
.
High protein diet from peanut can be produced from a byproduct from peanut oil production namely peanut meal
and its oil content ranging from about 1% to 7%, depending on the oil extraction method. Common method for oil
extraction is solvent extraction using hexane. However, residual solvent that cause toxicity issue on peanut meal has
become the limitation for this extraction method for food application. Physical method such as pressing method is
more environmentally safe to be implemented for the production of peanut meal but the output is still high in oil
content which can lead to oxidation and cause unpleasant odor. Therefore, another extraction method needs to
develop to produce meal with low oil content and no solvent residue.
Supercritical fluid carbon dioxide extraction (SCFE-CO
2
) has been found to be an alternative extraction
method
10-12
. This method has several advantages such as simple separation of the carbon dioxide (CO
2
) from the
extract and leaving no solvent residue in the extract. Unlike liquid solvents, the solving power of supercritical CO
2
can be easily adjusted with slight changes in the temperature and pressure, making it possible to extract particular
compounds of interest. In addition, CO
2
is inexpensive and available in high purity. CO
2
is also desirable for
compounds that are sensitive to extreme conditions because it has a relatively low critical point, with a critical
temperature (Tc) of 31.1C and a critical pressure (Pc) of 7.38 Mpa
13-15
. The efficiency of SCFE-CO
2
is determined
by various parameters such as extraction pressure, extraction temperature and CO
2
flow rate. The optimum
extraction condition can be achieved by investigating multiple factors in all possible conditions.
This present study investigated the application of supercritical fluid carbon dioxide extraction (SCFE-CO
2
) for
removing oil from black-eyed pea (Vigna unguiculata) and peanut (Arachis hypogaea) for having high protein-low
fat diet products. The objective of this study is to obtain the optimum process condition for high yield of oil extracts
from two seeds of Fabaceae family by SCFE-CO
2
using Taguchi method. Orthogonal array design (OAD) in
Taguchi method is a type of fractional factorial design that has been applied in several studies
14-19
. This approach
helps to identify the influence of individual factors and establish the relationship between variables and operational
conditions
20
. Taguchi method could also reduce and simplify experiments with large number of parameters and
levels with special orthogonal array design. This design determines configuration of each run with total number of
run, depending on the number of parameters and levels that are used.
Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272 267
2. Material and Method
2.1 Material
The peanut and black-eyed pea seeds (removed from pod and dried) were purchased from local farmer in Bogor
(West Java, Indonesia). Prior to extraction process, both seeds were grounded in disc mill (FFC-15, China) equipped
with 2 mm stainless steel round sieve. The grounded material then passed through 600 m stainless steel sieve for
peanut seed or 250 m stainless steel sieve for black-eyed pea seed. Liquid carbon dioxide (purity of 99.95%) was
supplied by PT Inter Gas Mandiri (Bekasi, West Java, Indonesia).
2.2 Method
2.2.1 Supercritical Carbon Dioxide Extraction
The supercritical fluid carbon dioxide extraction (SCFE-CO2) was perfomed using a customized supercritical
fluid apparatus described in the previous study
13
. Fig. 1 shows a schematic diagram of the system. Details of the
apparatus and experimental procedures were described in the previous paper
13
. Fifty grams of peanuts or 20 g of
black-eyed peas were used for each experiment. Extraction pressure was varied from 25 to 35 MPa and temperature
varied from 40 to 60C. The flow rate of CO
2
was varied from 10 to 20 g/min. The static stage was 60 min for all
experiments, while the dynamic stage was 180 min and 240 min, for peanut and black-eyed pea, respectively.
Fig. 1. Schematic diagram of supercritical CO
2
extraction system: 1. CO
2
Feed Tank; 2. CO
2
cylinder; 3. Condenser; 4. CO
2
pump;
5. CO
2
Pre-Heater; 6. Extractor; 7. Back Pressure Regulator; 8. Separator; 9. Absorber
2.2.2 Design of Experiments (DOE) and Statistical Analysis
Taguchi method L
9
orthogonal array was used in this experiment. Pressure, temperature and CO
2
flow rate were
used as parameters and each parameters held in three levels as shown in Table 1. The completed configuration of
Taguchi methods with L
9
array for supercritical CO
2
extraction of black-eyed pea and peanut is shown in Tables 2
and 3.
268 Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272
Table 1. Parameters and levels for supercritical CO
2
extraction of black-eyed pea and peanut
Parameters Level 1 Level 2 Level 3
Pressure (MPa) 25 30 35
Temperature (C) 40 50 60
Q-CO
2
(g/min) 10 15 20
The statistical analysis of the data obtained was performed in the form of analysis of variance (ANOVA) and
signal-to-noise (S/N) ratio calculation. ANOVA was used to determine the significant difference among the yield of
extraction. A probability of P<0.05 was considered significant with 95% confidence interval. MINITAB v.15
(Minitab Inc., USA) was used for all statistical analysis of the data obtained.
3. Result and Discussion
Nine experiments were performed for each peanut and black-eyed pea seeds based on standard L
9
orthogonal
arrays. Results of experiments in various conditions of black-eyed pea and peanut extraction are shown in Tables 2
and 3. Oil yield at each run of SCFE-CO
2
process is calculated by dividing weight of oil to the weight of raw
material. In this study, a light yellow transparent oil with the characteristic scent was obtained. The peanut and seeds
after SCFE-CO
2
process became a brittle chalky white color which indicated less or no oil content remain in the raw
material.
Table 2. Result of peanut extraction using supercritical CO
2
Run P T Q-CO
2 Time (min) SF Yield
(MPa) (C) (g/min)
Static Dynamic
(%)
1 25 40 10 60 180 180 38.36
2 25 50 15 60 180 270 42.48
3 25 60 20 60 180 360 45.25
4 30 40 15 60 180 270 46.94
5 30 50 20 60 180 360 44.37
6 30 60 10 60 180 180 45.43
7 35 40 20 60 180 360 45.73
8 35 50 10 60 180 180 48.19
9 35 60 15 60 180 270 48.61
Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272 269
Table 3. Result of black-eyed pea extraction using supercritical CO
2
Run P T Q-CO
2 Time (min) SF Yield
(MPa) (C) (g/min)
Static Dynamic
(%)
1 25 40 10 60 240 120 4.31
2 25 50 15 60 240 180 3.47
3 25 60 20 60 240 240 4.59
4 30 40 15 60 240 180 1.23
5 30 50 20 60 240 240 2.33
6 30 60 10 60 240 120 4.97
7 35 40 20 60 240 240 2.52
8 35 50 10 60 240 120 4.42
9 35 60 15 60 240 180 5.60
Fig. 2 shows the main effect plot of the extraction process to oil yield. Optimum condition of each parameter was
shown as the highest point of plot. Maximum yield of peanut oil extraction was achieved at pressure of 35 MPa,
temperature of 60
o
C and CO
2
flow rate of 15 g/min. Whereas the optimum conditions for black-eyed pea oil
extraction is achieved at extraction pressure of 25 MPa, temperature of 60
o
C and CO
2
flow rate of 10 g/min. In
addition to main effect plot, the signal-to-noise (S/N) ratio analysis was used to measure the quality of results. S/N
ratio with larger is better goal was chosen to optimize the process. Table 4 shows the result of S/N ratio analysis.
S/N ratio analysis revealed that the optimum condition for peanut oil extraction and black-eyed pea oil extraction is
similar with the optimum condition obtained using main effect plot (at 35 MPa, 60
o
C, 15 g/min for peanut oil
extraction and at 25 MPa, 60
o
C, 10 g/min for black-eyed pea oil extraction). S/N ratio analysis showed that pressure
is the highest influence parameter on oil yield in the supercritical extraction of peanut whereas temperature is the
highest influence parameter on oil yield in the supercritical extraction of black-eyed pea.
Fig. 2. Main effect plot for yield of black-eyed pea and peanut oil
270 Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272
Table 4. S/N Ratios calculation of peanut and black-eyed pea
Level P T Q-CO
2
Level P T Q-CO
2
1 32.45 32.77 32.83 1 12.24 7.49 13.17
2 33.17 33.05 33.24 2 7.69 10.36 9.18
3 33.53 33.33 33.09 3 11.96 14.04 9.54

1.08 0.56 0.42

4.55 6.55 3.99
Rank 1 2 3 Rank 2 1 3
Analysis of variance (ANOVA) was carried out to evaluate the statistical significance of pressure, temperature
and CO
2
flow rate on the amount of black-eyed pea and peanut oil extracted using supercritical CO
2
extraction. By
ANOVA, pressure has the greater effect than those of temperature and CO
2
flow rate on the yield of peanut oil
extraction. While on black-eyed pea oil extraction, temperature has the greatest effect (Tables 5 and 6). However,
the statistical analysis shows that there is a slight tendency not to be significant at confidence interval at 95%.
Table 5. ANOVA for S/N ratios of black-eyed pea
Source DF Seq SS Asj SS Adj MS F P
P 2 39.134 19.567 19.567 4.44 0.184
T 2 64.734 32.367 32.367 7.35 0.120
Q-CO
2
2 29.201 14.600 14.600 3.31 0.232
Residual Error 2 8.809 8.809 4.405
Total 8 141.878

Table 6. ANOVA for S/N ratios of peanut
Source DF Seq SS Asj SS Adj MS F P
P 2 1.8197 1.8197 0.9098 3.04 0.248
T 2 0.4711 0.4711 0.2356 0.79 0.560
Q-CO
2
2 0.2637 0.2637 0.1318 0.44 0.694
Residual Error 2 0.5989 0.5989 0.2994
Total 8 3.1534

Optimum condition obtained from means calculation and Taguchi method must be evaluated through real
experiment to check the accuracy of proposed optimum condition. Confirmation experiment was performed using
the proposed optimum conditions from Taguchi method. As a result, the confirmation experiment confirmed the
accuracy of Taguchi method for supercritical CO
2
extraction oil yield of black-eyed pea and peanut (Table 7). Oil
yield extraction using supercritical CO
2
extraction is 48.53% for peanut and 5.40% for black-eyed pea. Zia-ul-haq et
al.
21
reported oil yield from black-eyed pea using solvent extraction in a soxhlet apparatus was 2.85%.
Mukhopadhyay
22
reported oil yield from peanut using mechanical pressing and solvent extraction was 38%. Similar
result was reported by Kroschwitz and Howe-grant
23
for oil yield from peanut using solvent extraction. Therefore,
oil yield for both peanut and black-eyed pea extraction using supercritical CO
2
extraction were higher than
conventional extraction methods. These reports suggest that supercritical CO
2
extraction is an effective and efficient
method for oil extraction from black-eyed pea and peanut.
Karmelita Anggrianto et al. / Procedia Chemistry 9 ( 2014 ) 265 272 271
Table 7. Confirmation experiments at optimum conditions
Run Optimum P
(MPa)
T
(C)
Q-CO
2
(g/min)
Time (min) SF Yield
(%)
static dynamic
Peanut 35 60 15 60 180 270 48.53
Black-eyed pea 25 60 10 60 240 120 5.40
4. Conclusion
The optimization of supercritical carbon dioxide extraction (SCFE-CO
2
) of black-eyed pea and peanut seeds with
three parameters (pressure, temperature and CO
2
flowrate) with three levels has been performed using L
9
array of
Taguchi method to obtain the highest yield of oil. Based on the main effect plot and S/N ratio, the optimum
conditions for peanut extraction with yield of 48.53% peanut oil were at 35 MPa, 60C and 15 g/min of CO
2
flowrate, while for black-eyed pea extraction the optimum condition with yield of 5.4% black-eyed pea oil were
obtained at 25 MPa, 60
o
C and 10 g/min of CO
2
flowrate. S/N ratio analysis showed that the most prominent effect in
peanut oil extraction is pressure whereas in black-eyed pea is temperature, although from ANOVA test, it showed
that the effect is not significant (95% confidence level). Supercritical extraction can be used as alternative extraction
method to remove oil content from high-protein diet source for producing healthy food with low calorie and fat
content.
Acknowledgements
Authors thank to Sherly Juliani and Edward Widjojokusumo for critical review on this manuscript. This research
was supported by PT. Dexa Medica, Indonesia.
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