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Industrial Crops and Products xxx (2014) xxxxxx

Contents lists available at ScienceDirect

Industrial Crops and Products

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

Extraction of tocopherol-enriched oils from Quinoa seeds

by supercritical uid extraction
Katarzyna Przygoda, Grazyna Wejnerowska
Department of Food Analytics and Environmental Protection, Faculty of Chemical Technology and Engineering, University of Technology and Life Sciences in
Bydgoszcz, Seminaryjna 3 St., 85-326 Bydgoszcz, Poland

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

Article history: The efciency of supercritical uid extraction (SFE) in the production of oil with a high concentration of
Received 10 February 2014 tocopherols (vitamin E), from seeds of Quinoa (Chenopodium Quinoa Willd.), was investigated. The effect of
Received in revised form 9 September 2014 the process parameters such as pressure, temperature and time of extraction on the total tocopherol yield
Accepted 21 September 2014
was studied. For this purpose, the response surface methodology (RSM) was applied. The optimal SFE con-
Available online xxx
ditions for the tocopherol-enriched oil extraction from Quinoa seeds were: extracting pressure 18.5 MPa,
extracting temperature 130 C and extracting time 180 min. These optimum conditions yielded in tocoph-
Keywords:
erol concentration of 336.0 mg/100 g of oil and tocopherols were more than four times concentrated than
Seed oil extraction
Supercritical carbon dioxide extraction,
in classical hexane extraction.
Tocopherols 2014 Elsevier B.V. All rights reserved.
Quinoa seeds

1. Introduction has a high concentration of natural antioxidants like -tocopherol

Quinoa (Chenopodium Quinoa Willd.) is a pseudocereal that has
been cultivated in the Andean region for thousands of years. Nowa-
days Quinoa, as a cereal plant, is mostly grown in South America
and the worlds largest producers and exporters of Quinoa seeds
are Bolivia and Peru. Proposed by the government of Bolivia and
receiving strong support from many Central and South American
countries, Quinoa has now been singled out by The Food and Agri-
cultural Organization of the United Nations (FAO) as a food with
high nutritive value, impressive biodiversity and an important
role to play in the achievement of food security worldwide (Bojanic
(ed.), 2011). Recently, there has been growing interest in a number
of countries (especially in Europe), initiating cultivation introduc-
tion and research work on Quinoa (Bhargava et al., 2006). The
American and European Test of Quinoa have yielded good results
and demonstrate the potential of Quinoa as a grain and fodder crop.
Quinoa grain is highly nutritious due to its outstanding pro-
tein quality and wide range of minerals and vitamins (Bhargava
et al., 2006). The proximate composition of Quinoa ranges from
10 to 18% of protein, from 54.1 to 64.2% of carbohydrates, from
2.4 to 3.65 of ash, from 2.1 to 4.9% of crude bre and from 4.5 to
8.75% of crude fat (Hirose et al., 2010; Vilche et al., 2003). Quinoa
oil is rich in essential fatty acids, like linoleate and linolenate and
Corresponding author. Tel.: +48 523749040.
E-mail address: grazyna@utp.edu.pl (G.. Wejnerowska).
and -tocopherol (Bhargava et al., 2006; Hirose et al., 2010; Koziol,
1990; Ruales and Nair, 1992). Content of vitamin E (-, -, - and -
tocopherols) in Quinoa seeds given in literature is rather divergent
and it is 4.65.9 mg/100 g (Coulter and Lorenz, 1990; Miranda et al.,
2010; Ruales and Nair, 1992). Vitamin E is considered to be one of
the most important natural antioxidants which has its source in
products of plant origin. The presence of tocopherols in vegetable
oils has an effect on their protection against unfavourable oxidative
processes (Lampi and Kamal-Eldin, 1998). Therefore, Quinoa oil can
be used as a valuable source of tocopherols. Due to the high quality
of its oil and the fact that some varieties show fat concentrations
up to 9.5%, Quinoa is considered as a potentially valuable new oil
crop (Bhargava et al., 2006).
The oil from Quinoa can be used in many industries such as
in cosmetic, pharmaceutical and food industries. Solvent extrac-
tion methods used to extract oils have many disadvantages such
as toxic solvents and rst of all, these methods are characterized
by low selectivity and can lead to loss of valuable components
because the process is conducted at high temperatures (Koziol,
1993). Supercritical uid extraction (SFE) is a method for extrac-
tion of valuable ingredients from natural substances with high
yield and quality, being studied widely as an alternative to con-
ventional extraction methods (Bernardo-Gil et al., 2011). Several
vegetable materials have been obtained by using SFE. Supercritical
carbon dioxide (SCO2 ) is the most preferred supercritical solvent in
food extractions because it is non-toxic, non-explosive, inexpen-
sive, separated easily and completely from the extract, having the

http://dx.doi.org/10.1016/j.indcrop.2014.09.038
0926-6690/ 2014 Elsevier B.V. All rights reserved.

Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
G Model
INDCRO-7534; No. of Pages 7 ARTICLE IN PRESS
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potential for selective extractions by varying the pressure and tem-
perature (Bernardo-Gil et al., 2011; Follegatti-Romero et al., 2009).
Therefore, it seems advisable to apply supercritical uid extrac-
tion for obtaining oil with enhanced levels of vitamin E from C.
Quinoa. Many reports on obtaining oil with a higher content of
vitamin E from various plant materials such as: Silybum mari-
anum, residual olive husk, olive tree leaves, melon and roselle seeds
are given in literature (Calvo et al., 1994; de Lukas et al., 2002;
Follegatti-Romero et al., 2009; Gracia et al., 2011; Hadolin et al.,
2001; Nyam et al., 2010). However, there is a lack of information
available in the literature on the Quinoa oil extraction using SFE.
In the present work, the efciency of supercritical uid extrac-
tion applied to obtain oil with a high concentration of vitamin
E, from Quinoa seeds, was investigated. The inuence of process
parameters (pressure, temperature, time) on the total yield of
extraction and vitamin E content were investigated. Response sur-
face methodology (RSM) was used to evaluate how the different
factors inuenced the variables.
2. Materials and methods
2.1. Materials
Quinoa seeds (C. Quinoa Willd.) were bought from a local shop
(country of origin Bolivia). Total humidity of seeds was determined
by Sartorius MA 30 and was equal to 9.8%. Size of seeds before
grinding was 2.5 mm (80.4%). Quinoa seeds were grinded in ZM
200 Retsch grinder. Size of grinded seeds was determined by per-
forming sieve analysis and it was 1.01.4 mm (68%).
2.2. Reagents and standards
Reference tocopherols, DL--T (>96%) from Sigma Chemicals,
rac--T(90+%), -T (99%) and -T (93.7%) were purchased from
Supelco (Bellefonte, PA, USA). Ethyl acetate and n-hexane were
HPLC grade and were obtained from POCH S.A. (Gliwice, Poland).
Carbon dioxide (99.5%) was obtained from Linde Gas (Poland).
The ne-grain sand for Soxhlet and SFE extractions was sieved
on sieves and fractions of 0.20.3 mm were collected. Then, the
sand was puried by successive elution with warm distilled water,
methanol and hexane. The sand was dried after each stage of elu-
tion.
2.3. Methods
2.3.1. Classical extraction
Samples of 10 g of ground Quinoa seeds were weighted with
accuracy of 0.0001 g and then were mixed with 10 g of sand to
determine the oil content by a Soxhlet extraction using n-hexane
at 60 C for 16 h. After extraction n-hexane was evaporated under
vacuum at 40 C and subsequently the solvent was totally removed
by nitrogen steam. After evaporation of solvent, the oil content was
determined gravimetrically. The mass of extracted oil was assumed
to be 100% of the extractable matter.
2.3.2. Supercritical uid extraction procedures
A laboratory-scale SFE system Lizard 2001 SEKO-K s.r.o (Brno,
Czech Republic) for maximum pressure of 40 MPa and a tempera-
ture of 150 C, was used in this study. The schematic apparatus used
for supercritical CO2 extraction is shown in Fig. 1. Ground seeds of
Quinoa were loaded into the extractor cell of 1.2 ml capacity (0.5 cm
internal diameter (I.D.) and 6.1 cm of effective height). About 0.4 g of
sand and 0.6 g of ground seeds (weighted with accuracy of 0.0001 g)
were located into the cell and the content of cell was stirred for
5 min by use of rotary stirrer. The extract was collected into 12 ml
vials (previously weighted). The experiments were carried out in
Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
Fig. 1. Schematic diagram of supercritical CO2 extraction apparatus of LIZARD 2001
SEKO-K. 1 CO2 cylinder; 2 onoff valve; 3 CO2 pump; 4 CO2 pump chiller;
5 cosolvent; 6 cosolvent pump; 7 extraction cell; 8 restrictor heating 9
trapping vial; 10 heating/cooling block.
temperature from 30 to 130 C, at pressure from 12.5 to 28.5 MPa
for 15180 min.
The adjustment of CO2 ow is not possible in this SFE system.
Measurements of CO2 ow rate were performed at the end of cap-
illary (restrictor) with diameter of 45 m and length of 7 cm. The
SCO2 ows were dependent on extraction conditions and they were
within the range from 10.5 to 27.0 l/h.
The extraction yield was determined by comparing the weight
of oil obtained by SFE with the weight of oil obtained by Soxhlet
extraction. Extracts obtained by SFE at the different conditions were
tested to determine concentration of tocopherols. All experimental
results (SFE and HPLC analysis) reported are average values from
three repeated independent experimental runs. Standard error
among the data was about 5%.
2.3.3. Determination of tocopherol concentrations in the extract
Content of tocopherols in the extracted oil samples was
determined using a high-performance liquid chromatography sys-
tem from Shimadzu (Kyoto, Japan). The chromatographic system
included a uorescence detector (ex = 294 nm, em = 330 nm) and
NH2 -NP (250 mm 4.6 mm, 5 m) column (Supelco, Bellefonte, PA,
USA). The eluent was n-hexane/ethyl acetate 70:30 (v/v) at constant
1 ml/min ow rate. Prior to HPLC analysis, the oils were diluted with
0.5 or 1.0 ml n-hexane, ltered (0.45 m nylon syringe lter) and a
20 l sample was injected.
3. Results and discussion
The aim of the studies was to obtain tocopherol-enriched oils
from Quinoa by supercritical uid extraction with CO2 . Total con-
tent of oil in Quinoa seeds was determined by weight using
Soxhlet method and it was 7.21 0.42 g/100 g of seeds. Concen-
trations of the individual tocopherols in the oil studied were
as follows: -tocopherol 22.74 0.40, -tocopherol 48.42 0.44,
-tocopherol 1.62 0.07 mg/100 g. Content of -tocopherol was
below 0.04 mg/100 g of oil. The sum of tocopherol homologues in
the oil obtained was 72.78 mg/100 g of oil.
As it is known, the parameters (pressure, temperature, SCO2
ow and extraction time) of the SFE process have a signicant
impact on the quantity of oil extracted from seeds, and most impor-
tantly on its composition. In the initial stage of research, the SFE
parameters were chosen on the basis of literature reports. Nyam
et al. (2010) investigated the possibilities of using Kalahari melon
and roselle seeds to obtain tocopherol-enriched oils. They obtained
the highest concentrations of tocopherols (274.74 mg/100 g of
oil) for Kalahari melon by SCO2 extraction under conditions:
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29 MPa; 58 C; 180 min, while for roselle seeds, the result was

Coded and uncoded matrix of the experiments and the extraction yields of Quinoa oils (g/100 g of seeds; %) and yields of tocopherols (mg/100 g of oil; %) using supercritical CO2 with different combinations of temperature ( C),

Tocopherol
89.75 mg/100 g of oil at optimum conditions: MPa; 80 C; 180 min.

yield (%)
Other researchers who have dealt with the determination of opti-

13.74
0.57
24.43
2.67
31.49
12.98
9.92
18.13
0.38
25.19
11.64
12.98
13.36
0.19
27.29
14.31
8.21
25.57
12.79
13.36
mal conditions for SCO2 extraction of tocopherols from residual
olive husk state that the best yields were achieved under the pro-
cess conditions: 25 MPa; 40 C; 360 min and the result obtained
was 15.79 mg of tocopherols in 100 g of olive husk oil (Gracia et al.,
2011). Hadolin et al. (2001) obtained vitamin E-rich oil from S.

(mg/100 g seeds)
Tocopherol yield
marianum and the most concentrated extract of -tocopherol was
achieved at 20 MPa; 80 C.
Mendiola et al. (2008) studied possibilities to obtain fractions
highly enriched in vitamin E from Spirulina platensis micro alga

0.72

1.28
0.14
1.65
0.68
0.52
0.95

1.32
0.61
0.68

1.43
0.75
0.43
1.34
0.67
0.03

0.02

0.70
0.01

0.70
by SFE and under conditions: 22 MPa, 83.3 C and 75 min. They
obtained the highest concentration of tocopherols in the material
studied. It results from these data that the extraction parameters

117.52
24.99
58.85
85.32
120.11

149.75
66.18

112.88
56.32
201.32

43.22
120.59
107.73
169.77
119.93
111.53
116.50

22.01
85.06

14.03
in the cases described were different.

Sum
No data concerning extraction of oil from Quinoa seeds by SFE
method have been found in literature. Taking into account the
literature data and the results of preliminary experiments con-

5.21
5.15

6.23
4.87
6.48
1.94
2.86
3.46
4.12
2.19

2.42
1.37
5.96
4.93
5.47
5.69
4.45
1.09

4.80
6.00
-T
ducted in our laboratory, we decided to carry out the extraction
in temperature range of 50100 C, at pressure of 12.524.5 MPa

Tocopherol concentration
and in time 3080 min. Response surface methodology (RSM) using

82.32
15.78
41.34
59.49
78.12
82.25

79.88
37.54
141.54

29.53
83.35
66.52
115.48

79.86
103.03
47.03
11.01
57.30

7.50

84.06
central composite design (CCD) was employed to determine the

(mg/100 g of oil)

-T
effect of three variables on the tocopherol concentration. Zas-
tosowalismy RSM poniewaz jest najbardziej znana i najczesciej
stosowana metoda do optymalizacji wielu procesw chemicznych

29.99

16.42
19.83
35.76
29.38
40.24
17.21
8.14

28.88
16.59
54.98
4.11
12.32
31.28
36.28
48.82
30.18
27.22
4.06

24.30
i biochemicznych. In our studies, 20 experiments were planned

-T
in which the impact of three independent parameters, namely,
pressure, temperature and extraction time on the amount of oil
obtained was determined. The computer program Statistica 6.0 was
Oil yield (%)

applied to experiment planning, analysis of experimental data and

graphical presentation of spatial response surface plots. The results 8.46

2.22

4.85
19.97
1.66

7.49
16.78
5.27
0.55
45.63

5.55
10.96
7.77
8.74
1.80
30.10

8.04

21.50

8.60
19.00
of studies were presented as the amount of oil obtained (g/100 g of
seeds) and tocopherol concentration in oil (mg/100 g). The yields of
the process for the oil obtained and tocopherols (%) were presented
in comparison with the results obtained by the Soxhlet method.
(g/100 g seeds)

The mean values of results were calculated from at least three rep-
etitions of experiments. Parameters of extraction and results of
Oil yield

studies are presented in Table 1.

0.61
0.13
2.17
0.16
1.37
0.58
0.35
1.44
0.12
1.55
0.54
1.21
0.38

3.29
0.62

0.79
0.56
0.63
0.04

0.40
Response surface plots showing the effect of extraction time,
temperature and pressure on tocopherol yield are presented in
Fig. 2(a)(c).
Time

55
30
80
80
30
55
80
30
30
80
55
55
55
55
55
55
15
100
55
55
Table 1 shows the tocopherol concentration in oil obtained
under the fteen different testing conditions (T, P, t). Variance anal-
ysis of the factors studied for the response surface model is given in
Actual parameter values

Pressure

Table 2. The mathematical model generated from the experimen-

tal data using Statistica 6.0 is expressed by the following quadratic
18.5
12.5
24.5
12.5
24.5
18.5
12.5
24.5
12.5
24.5
18.5
18.5
18.5
8.5
28.5
18.5
18.5
18.5
18.5
equation (Y is the total concentration of tocopherols in oil): 18.5

Y = 117.3088 + 18.7766 T 0.0967 T 2 + 7.1305

Temp.

75
50
50
100
100
75
50
50
100
100
75
35
120
75
75
75
75
75
75
75

P 35.8806 P 2 + 18.3458 t + 3.4497 t 2 + 18.4437

T P 11.1462 T t 28.8063 P t
1.68
1.68
0.0
1.0
1.0
1.0
1.0
0.0
1.0
1.0
1.0
1.0
0.0
0.0
0.0
0.0
0.0
0.0

0.0
0.0
t

Comparison of tocopherol concentrations in oil, actual values

and predicted values are presented in Fig. 3. The second order
pressure (MPa) and time (min).

1.68
1.68
Coded parameter

polynomial equation model was in quite good agreement with the

0.0
1.0
1.0
1.0
1.0
0.0
1.0
1.0
1.0
1.0
0.0
0.0
0.0

0.0
0.0
0.0
0.0
0.0

experimental results, with R2 of 0.84.

P

The response surface plot was plotted with a single variable

holding constant at zero (zero-level) and the rest of the extrac-
1.68
1.68
0.0
1.0
1.0
1.0
1.0
0.0
1.0
1.0
1.0
1.0
0.0

0.0
0.0
0.0
0.0
0.0
0.0
0.0

tion parameters changed in the ranges of the experiment. The zero

T

values for experiment were 75 C, 18.5 MPa, and 55 min for temper-
ature, pressure and time, respectively. It can be observed from the
Table 1

Run

results obtained that pressure of extraction is an important param-

11
12
13
14
15
16
17
18
19
10

20
1
2
3
4
5
6
7
8
9

eter independently of temperature and time of extraction. On the

Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
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Fig. 2. Response surface plots of SCO2 for tocopherol yields (mg tocopherols/100 g of oil): (a) effect of pressure and temperature at 55 min; (b) effect of time and pressure at
75 C; (c) effect of time and temperature at 18.5 MPa.

Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
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Table 2
ANOVA table of varieties.

Sourcea Sum of squares df Mean square F value p-Valueb

Model 39,470.4 9 4385.6 5.96 <0.0001

T 4794.8 1 4794.8 6.52 0.0287
T2 0.13 1 0.13 1.8 104 0.9895
P 691.5 1 691.5 0.94 0.3550
P2 18,275.9 1 18,275.9 24.86 0.0005
t 4577.3 1 4577.3 6.22 0.0317
t2 168.9 1 168.9 0.22 0.6420
TP 2721.4 1 2721.4 3.70 0.0833
Tt 993.9 1 993.9 1.35 0.2720
Pt 6638.4 1 6638.4 9.03 0.0132
Residual 7352.8 10 735.8
Lack of t 7285.4 5 1457.1 108.14 <0.0001
Pure error 67.4 5 13.5
Corrected total 46,823.1 19
a
Pressure (P), temperature (T), time (t).
b
P < 0.05 indicates statistical signicance.

basis of Fig. 1a and b, presenting an effect of pressure, it was found
that the highest concentration of tocopherols was obtained for
18.5 MPa. Extraction performed both at lower and higher pressures
resulted in lower extraction yields. Moreover, it can be concluded
from the results obtained (Table 1) that when extraction was con-
ducted at time 5580 min and temperature 5075 C, the increase
in pressure had unfavourable effect on the content of tocopherols
in the extract. Simultaneously with the increase of pressure, the
amount of tocopherols (g/100 g seeds) extracted under conditions
of high temperature (100 C) and long time of extraction (80 min)
increases.
Effect of temperature of extraction on concentration of toco-
pherols was presented in Fig. 1a and c. The temperature range in
our research plan was within the range from 35 to 120 C. The
extraction yield increases with temperature rise at constant time
(Fig. 1a) and constant pressure (Fig. 1c). These results are in agree-
ment with results of other researchers who described the similar
dependences.
Effect of time of extraction on concentration of tocopherols was
presented in Fig. 1b and c and it was observed that similarly as for
temperature, concentration of tocopherols in a sample increased
with the longer time of extraction. The highest content of total
tocopherols i.e. 201.3 mg/100 g of oil was obtained at temperature
of 120 C (18.5 MPa; 55 min) (Table 3).
The further studies allowed us to formulate more precise
conclusions concerning the effect of parameters on efciency of
tocopherol SCO2 extraction from Quinoa seeds. The content of
tocopherols in the extracts increases very signicantly at high tem-
peratures and at lower pressures (130 C, 18.5 MPa). A contrary
dependence is observed in the case of oil yield which is the most
favourable at lower temperatures and at high pressures (75 C,
24.5 MPa). Similar dependences were observed by Hadolin et al.
(2001). They stated that increase in pressure during extraction
reduced concentration of tocopherols in the extract.
It was observed that increase in extraction pressure had a
favourable effect on concentration of tocopherols (mg/100 g of oil)
at short time of extraction (30 min), both at low 50 C and high
120 C temperature. However, temperature rise to 130 C results in
decrease in concentration of tocopherols in oil. Effect of pressure,
time and temperature of extraction on tocopherol content in the
extract is presented in Fig. 4.
Independently of the pressure and temperature conditions,
longer time of extraction has a positive effect on the amount both
of oil and tocopherol extracted. Extraction conducted for 60 min
(18.5 MPa; 130 C) (Table 2) caused the increase in tocopherol con-
tent in the extract to 299.5 mg tocopherols/100 g of oil. Only longer
time of extraction could lead to higher yield what was veried
in further studies. It was decided to extend the extraction time
to 180 min. Parameters of extraction and the results obtained are
presented in Table 4.
As expected, longer time of extraction resulted in increase in the
amount of tocopherols extracted. The most preferred results were
obtained when the extraction was carried out for 180 min, which
resulted in obtaining the oil containing tocopherols in amount of
336.0 mg/100 g of oil. Extraction conducted under the selected con-
ditions (18.5 MPa; 130 C; 180 min) allowed us to obtain tocopherol

Fig. 3. Comparison of actual values and predicted values of tocopherols concentra- Fig. 4. Effect of pressure, time and temperature of extraction on tocopherol concen-
tion in oil. Predicted values were calculated from equation. tration in the extract (mg tocopherols/100 g of oil).

Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
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Table 3
The extraction yields of Quinoa oils (g/100 g of seeds; %) and yields of tocopherols (mg/100 g of oil; %) using supercritical CO2 with different combinations of temperature
( C), pressure (MPa) and time (min).

Run Actual parameter values Oil yield Oil yield % Tocopherol concentration Tocopherol yield Tocopherol
g/100 g seeds mg/100 g of oil mg/100 g seeds yield%

Temperature Pressure Time -T -T -T Sum

1 110 24.5 30 0.86 11.93 31.57 90.89 4.80 127.26 1.09 20.96
2 110 24.5 60 1.07 14.84 45.12 107.21 3.46 155.79 1.66 31.92
3 110 18.5 30 0.27 3.74 32.40 86.33 4.57 123.30 0.33 6.35
4 110 18.5 60 0.30 4.16 108.95 128.23 8.33 245.51 0.74 14.23
5 120 24.5 30 0.58 8.04 57.81 142.64 7.29 207.74 1.21 23.27
6 120 24.5 60 0.80 11.10 55.66 122.39 7.26 185.31 1.49 28.65
7 120 18.5 30 0.21 2.91 44.87 121.54 7.39 173.80 0.36 6.92
8 120 18.5 60 0.34 4.72 46.84 111.43 6.43 164.70 0.56 10.77
9 130 24.5 30 0.40 5.55 38.00 128.78 8.09 174.87 0.69 13.27
10 130 24.5 60 0.47 6.52 64.48 142.22 9.35 216.05 0.99 19.04
11 130 18.5 30 0.29 4.02 39.94 91.11 6.52 137.57 0.40 7.69
12 130 18.5 60 0.31 4.30 86.13 193.09 20.31 299.53 0.93 17.88

Table 4
Effect of time on the yield of tocopherol extraction from Quinoa seeds.

Run Actual parameter values Oil yield (g/100 g Oil yield (%) Tocopherol concentration Tocopherol yield Tocopherol
seeds) (mg/100 g of oil) (mg/100 g seeds) yield (%)

Temp. Pressure Time -T -T -T Sum

1 130 18.5 90 0.33 4.58 88.91 202.04 17.13 308.08 1.02 19.96
2 130 18.5 120 0.35 4.85 92.19 207.43 20.07 319.69 1.12 21.54
3 130 18.5 180 0.40 5.54 109.08 208.82 18.10 336.00 1.34 25.77

Table 5
Comparison the Soxhlet extraction with SFE at 130 C/18.5 MPa.

Extraction methods Tocopherol concentration (mg/100 g of oil) Tocopherol yield (mg/100 g seed) Tocopherol yield (%)

-T -T -T Total

Soxhlet extraction 22.74 48.42 1.62 72.78 5.24 100.0

SFE (130 C; 18.5 MPa; 60 min) 86.13 193.09 20.31 299.53 0.93 17.9
SFE (130 C; 18.5 MPa; 180 min) 109.08 208.82 18.10 336.00 1.34 25.8

Signicantly lower values (2838%) for tocopherol-enriched oils

were obtained for Kalahari melon, roselle seeds and Sacha inchi
(Follegatti-Romero et al., 2009; Nyam et al., 2010). However, other
researches who obtained vitamin E-rich oil from S. marianum, resid-
ual olive husk and sunower seeds achieved signicant increase
(98300%) in tocopherol concentration in oil (de Lukas et al., 2002;
Gracia et al., 2011; Hadolin et al., 2001).

4. Conclusion

In this work, we have used supercritical carbon dioxide for the

Fig. 5. Effect of extraction time on the yield (18.5 MPa; 130 C).
yield of 25%. Effect of time on the yield of tocopherol extraction from
Quinoa seeds is presented in Fig. 5.
Prolongation of reaction time from 60 min to 180 min resulted
in increase in the yield of tocopherol extraction from seeds by
about 44% (from 0.93 to 1.34%). Moreover, a signicant increase
in tocopherol concentration in the oil obtained from Quinoa seeds
was observed. Concentration of tocopherols in oil obtained by
Soxhlet method was 72.8 mg/100 g of oil, while the content of toco-
pherols in the extract obtained by SFE was 336.0 mg/100 g of oil, i.e.
tocopherol concentration in the extract increased by about 462%.
Such a result was obtained for the following extraction conditions:
130 C, 18.5 MPa and 180 min extraction. Comparison of the results
obtained by SFE and Soxhlet extraction is presented in Table 5.
extraction oil with high concentration of tocopherols (vitamin E)
from Quinoa seeds. Results were compared with those obtained
for conventional extraction with organic solvent (n-hexane).
Supercritical CO2 extraction was conducted at temperatures
of 30130 C; pressures of 12.528.5 MPa; times 15180 min and
CO2 ow rate of 10.527.0 l/h. Application of response surface
methodology (RSM) with central composite design (CCD) enabled
to optimize SFE parameters. As a result of our studies, it was found
that the extract with high content of tocopherols was obtained
at high temperatures (120130 C) and at pressure of 1820 MPa.
Independently of the pressure and temperature conditions, longer
time of extraction has a positive effect on the amount both of oil and
tocopherol extracted. The optimum conditions were: extracting
pressure 18.5 MPa, extracting temperature 130 C and extracting
time 180 min. The yield of tocopherol extraction under optimum
conditions is relatively high and it is 25.65%, the total content of
tocopherols in the extract is 336.0 mg/100 g. The results indicate

Please cite this article in press as: Przygoda, K., Wejnerowska, G.., Extraction of tocopherol-enriched oils from Quinoa seeds by super-
critical uid extraction. Ind. Crops Prod. (2014), http://dx.doi.org/10.1016/j.indcrop.2014.09.038
G Model
INDCRO-7534; No. of Pages 7 ARTICLE IN PRESS
K. Przygoda, G.. Wejnerowska / Industrial Crops and Products xxx (2014) xxxxxx
that tocopherols are more than four times concentrated than those
obtained by classical hexane extraction.
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