Application of Supercritical Fluid Extraction on Food Processing: Black-eyed Pea (Vigna Unguiculata) and Peanut (Arachis Hypogaea)

Available online at www.sciencedirect.com

ScienceDirect Procedia Chemistry 9 (2014) 265 – 272

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-CO2) for removing fats from black-eyed pea (Vigna unguiculata) and peanut (Arachis hypogaea) to produce high protein-low fat diet products. SCFE-CO2 process used Taguchi’s orthogonal array at three parameters in three levels: pressures between 25-35 MPa, temperatures between 40-60oC and CO2 flowrates between 10-20 g/min. The results of the experiment showed that the optimum conditions of SCFECO2 process were 25 MPa, 60oC, 10 g/min for black-eyed pea and 35 MPa, 60oC, 15 g/min for peanut. The highest yield for black-eyed pea and peanut were 5.4% and 48.5%, respectively. © Authors. et Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license © 2014 2014The Anggrianto al. Published by Elsevier B.V. (http://creativecommons.org/licenses/by-nc-nd/3.0/). Selection and peer-review under responsibility of the Organizing Committee of ICCE UNPAR 2013. 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 Adj SS DF Pc Q Seq SS

adjusted mean of square adjusted sum of square degree of freedom critical pressure (MPa) flow rate sequenced sum of square

* Corresponding author. Tel.: +62-21-898-41901; fax: +62-21-898-41905 . E-mail address: [email protected].

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

266

SF S/N Tc

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solvent/feed ratio signal-to-noise critical temperature (ΣC)

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 disease1-3. 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 countries4. Peanut contains approximately 38% oil w/w5 and 25-28 % w/w of protein6. Black-eyed pea (Vigna unguiculata) is another important grain legumes which forms the source of dietary protein7,8. The black-eyed pea protein content is relatively high between 19.5 % and 27.3 % w/w9. 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 tonic7. 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-CO2) has been found to be an alternative extraction method10-12. This method has several advantages such as simple separation of the carbon dioxide (CO2) from the extract and leaving no solvent residue in the extract. Unlike liquid solvents, the solving power of supercritical CO2 can be easily adjusted with slight changes in the temperature and pressure, making it possible to extract particular compounds of interest. In addition, CO2 is inexpensive and available in high purity. CO2 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 31.1°C and a critical pressure (Pc) of 7.38 Mpa13-15. The efficiency of SCFE-CO2 is determined by various parameters such as extraction pressure, extraction temperature and CO2 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-CO2) 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-CO2 using Taguchi method. Orthogonal array design (OAD) in Taguchi method is a type of fractional factorial design that has been applied in several studies14-19. This approach helps to identify the influence of individual factors and establish the relationship between variables and operational conditions20. 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.

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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 µ m stainless steel sieve for peanut seed or 250 µ 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 study13. Fig. 1 shows a schematic diagram of the system. Details of the apparatus and experimental procedures were described in the previous paper13. 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 60ΣC. The flow rate of CO2 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 CO2 extraction system: 1. CO2 Feed Tank; 2. CO2 cylinder; 3. Condenser; 4. CO2 pump; 5. CO2 Pre-Heater; 6. Extractor; 7. Back Pressure Regulator; 8. Separator; 9. Absorber

2.2.2 Design of Experiments (DOE) and Statistical Analysis Taguchi method L9 orthogonal array was used in this experiment. Pressure, temperature and CO2 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 L9 array for supercritical CO2 extraction of black-eyed pea and peanut is shown in Tables 2 and 3.

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Karmelita Anggrianto et al. / Procedia Chemistry 9 (2014) 265 – 272 Table 1. Parameters and levels for supercritical CO2 extraction of black-eyed pea and peanut Parameters

Level 1

Level 2

Level 3

Pressure (MPa)

25

30

35

Temperature (ιC)

40

50

60

Q-CO2 (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 L9 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-CO2 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-CO2 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 CO2 T

Q-CO2

(MPa)

(϶C)

(g/min)

1

25

40

2

25

3

Run

P

Time (min)

SF

Yield (%)

Static

Dynamic

10

60

180

180

38.36

50

15

60

180

270

42.48

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 Table 3. Result of black-eyed pea extraction using supercritical CO2 P (MPa)

T (϶C)

Q-CO2 (g/min)

Time (min) Static

Dynamic

1

25

40

10

60

2

25

50

15

3

25

60

20

4

30

40

5

30

6

30

7 8 9

Run

SF

Yield (%)

240

120

4.31

60

240

180

3.47

60

240

240

4.59

15

60

240

180

1.23

50

20

60

240

240

2.33

60

10

60

240

120

4.97

35

40

20

60

240

240

2.52

35

50

10

60

240

120

4.42

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 60oC and CO2 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 60oC and CO2 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, 60oC, 15 g/min for peanut oil extraction and at 25 MPa, 60oC, 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

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Table 4. S/N Ratios calculation of peanut and black-eyed pea Level P Level P T Q-CO2

T

Q-CO2

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

ȴ

1.08

0.56

0.42

ȴ

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 CO2 flow rate on the amount of black-eyed pea and peanut oil extracted using supercritical CO2 extraction. By ANOVA, pressure has the greater effect than those of temperature and CO2 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

3.31

0.232

Q-CO2

2

29.201

14.600

14.600

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

0.44

0.694

Q-CO2

2

0.2637

0.2637

0.1318

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 CO2 extraction oil yield of black-eyed pea and peanut (Table 7). Oil yield extraction using supercritical CO2 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%. Mukhopadhyay22 reported oil yield from peanut using mechanical pressing and solvent extraction was 38%. Similar result was reported by Kroschwitz and Howe-grant23 for oil yield from peanut using solvent extraction. Therefore, oil yield for both peanut and black-eyed pea extraction using supercritical CO2 extraction were higher than conventional extraction methods. These reports suggest that supercritical CO2 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

Table 7. Confirmation experiments at optimum conditions P (MPa)

T (ΣC)

Peanut

35

Black-eyed pea

25

Run Optimum

Q-CO2 (g/min)

Time (min) static

dynamic

60

15

60

60

10

60

SF

Yield (%)

180

270

48.53

240

120

5.40

4. Conclusion The optimization of supercritical carbon dioxide extraction (SCFE-CO2) of black-eyed pea and peanut seeds with three parameters (pressure, temperature and CO2 flowrate) with three levels has been performed using L9 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, 60ΣC and 15 g/min of CO2 flowrate, while for black-eyed pea extraction the optimum condition with yield of 5.4% black-eyed pea oil were obtained at 25 MPa, 60oC and 10 g/min of CO2 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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