Академический Документы
Профессиональный Документы
Культура Документы
Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem
a r t i c l e
i n f o
Article history:
Received 23 May 2012
Received in revised form 13 September 2012
Accepted 20 September 2012
Available online 29 September 2012
Keywords:
Pulsed electric elds
Extraction
Anthocyanin
Purple-eshed potato
a b s t r a c t
The inuence of pulsed electric eld (PEF) treatment on the anthocyanin extraction yield (AEY) from purple-eshed potato (PFP) at different extraction times (60480 min) and temperatures (1040 C) using
water and ethanol (48% and 96%) as solvents has been investigated. Response surface methodology
was used to determine optimal PEF treatment and optimise anthocyanin extraction. A PEF treatment
of 3.4 kV/cm and 105 ls (35 pulses of 3 ls) resulted in the highest cell disintegration index (Zp = 1) at
the lowest specic energy requirements (8.92 kJ/kg). This PEF treatment increased the AEY, the effect
being higher at lower extraction temperature with water as solvent. After 480 min at 40 C, the AEY
obtained for the untreated sample using 96% ethanol as the solvent (63.9 mg/100 g fw) was similar to
that obtained in the PEF-treated sample using water (65.8 mg/100 g fw). Therefore, PEF was possible with
water, a more environmental-friendly solvent than ethanol, without decreasing the AEY from PFP.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Colour is recognised as a major factor affecting food acceptance.
Enhancing food colour during processing by adding colorants is a
common practice in the food industry to make food more appealing for consumers. In recent years, there has been an increasing
interest in the food industry toward replacing synthetic colorants
by natural ones, as a consequence of the doubts about the safety
of synthetic colorants and the consumer demand for natural products (Sowbhagya & Chitra, 2010). Anthocyanins, a group of phenolic compounds, are some of the most extensive natural pigments,
responsible for the red, purple and blue colours of fruits, vegetables
and owers (Mazza & Miniati, 1993). Interest in anthocyanin pigments has increased recently, due to the range of colours of their
molecules, with their possible applications as natural dyes and also
potential health benets as dietary antioxidants (He & Giusti,
2010; Lachman et al., 2009; Suda et al., 2003).
Pigmented potato cultivars such as purple-eshed potatoes
(PFPs) are a rich source of anthocyanins. It has been reported that
the concentration of anthocyanins in PFP is similar to the highest
anthocyanin production crops, such as blueberries, blackberries,
cranberries or grapes (Bridgers, Chinn, & Truong, 2010). Due to
the fact that they are a low-cost crop, purple-eshed potatoes
may be a potential source of natural anthocyanin pigments for
industry (Jansen & Flamme, 2006).
0308-8146/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2012.09.080
Studies conducted on the extraction of anthocyanins from different plants have showed that extraction yield depends on different factors, such as solvent type and concentration or time and
temperature of extraction (Cacace & Mazza, 2003). Anthocyanins
are soluble in water; however, extraction methods generally used
for obtaining anthocyanin pigments from plants are usually based
on the use of other polar solvents, such as methanol or ethanol. For
example, Bridgers et al. (2010) observed that ethanol and methanol
extracts of PFP had approximately 34 times higher values of
anthocyanins compared to water extracts. However, the use of
these kinds of solvents increases the cost of the process and may
cause important environmental problems. Consequently, improving the anthocyanin extraction with water by the use of pretreatment steps, such as application of pulsed electric elds (PEFs)
is an interesting approach.
PEF is an emerging technology that has gained increasing interest in recent years for improving mass transfer operations in the
food industry (Donsi, Ferrari, & Pataro, 2010; Knorr et al., 2011;
Purtolas, Luengo, lvarez, & Raso, 2012). The process is based
on the application of external electric elds that induce the electroporation of eukaryotic cell membranes, enhancing the diffusion of
solutes. This permeabilisation of cell membranes can be achieved
at moderate electric elds (<10 kV/cm) and low specic energies
(<10 kJ/kg).
An enhancement in the extraction of colorants, such as anthocyanins from grapes or red cabbage (Corrales, Toep, Butz, Knorr, &
Tauscher, 2008; Gaschovska et al., 2010) or betanin from red beets
(Chalermchat, Fincan, & Djmek, 2004; Fincan, DeVito, & Dejmek,
2004; Lpez, Purtolas, Condn, Raso, & lvarez, 2009), by application of a PEF treatment has been reported. However, these studies
only evaluated the aqueous extraction of these compounds.
1331
Zp 1
K h K 0h K 0l
; 0 6 Zp 6 1
K 0h K h K l
where Kl, K 0l are the electrical conductivities of untreated and treated material, respectively, in a low-frequency eld (15 kHz); Kh, K 0h
are the electrical conductivities of untreated and treated material,
respectively, in a high-frequency eld (350 MHz). The Zp varies between 0 for intact tissues and 1 for a tissue with all the cells
permeabilised.
2.4. Anthocyanin extraction
PEF equipment used in this investigation was supplied by ScandiNova (Modulator PG; ScandiNova, Uppsala, Sweden). The apparatus generates square waveform pulses of a width of 3 ls with a
frequency up to 300 Hz. The maximum output voltage and current
were 30 kV and 200 A, respectively. The equipment consists of a direct current power supply which converts the 3-phase line voltage
to a regulated DC voltage. It charges up 6 IGBT switching modules
(high-power solid-state switches) to a primary voltage of around
1000 V. An external trigger pulse gates all the modules and controls its discharge to a primary pulsed signal of around 1000 V. Finally, a pulse transformer converts this primary 1000 V pulse to
the desired high-voltage pulse.
The treatment chamber consisted of a cylindrical methacrylate
tube closed with two polished stainless steel cylinders. The gap between the electrodes was 2 cm. The diameter of the treatment
chamber was 2 cm for the determination of the cell disintegration
index, and 3.4 cm for the extraction experiments.
Actual voltage and current intensity applied were measured
with a high-voltage probe (P6015A; Tektronix, Wilsonville, OR)
and a current probe (Stangenes Industries Inc., Palo Alto, CA),
respectively, connected to an oscilloscope (TDS 220, Tektronix).
PEF treatments ranging from 5 to 35 pulses of 3 ls (45105 ls),
set at electric eld strength ranging from 1 to 5 kV/cm were used.
Specic energy of these treatments ranged from 0.54 to 13.50 kJ/kg
(Table 1). A pulse frequency of 1 Hz was used.
Table 1
Cell disintegration index (Zp) of purple-eshed potato (PFP) after application of PEF treatments at different electric eld strengths and treatment times. Specic energy for the
different treatments conditions has been also included.
Zp*
1
2
2
3
3
3
4
4
5
75
45
105
15
75
145
45
105
75
0.54
1.26
3.02
0.97
4.86
8.75
5.20
12.1
13.5
0.19 0.03
0.12 0.02
0.53 0.11
0.00 0.00
0.83 0.04
1.00 0.01
0.80 0.02
0.98 0.03
1.00 0.01
1332
The CCD and the corresponding analysis of the data were carried out using the software package Design-Expert 6.0.6 (Stat-Ease
Inc., Minneapolis, MN).
Y b0
k
k
k
X
X
X
bi X i
bii X 2i
bij X i X j
i1
i1
i>j
where Y is the response variable to be modelled, Xi and Xj are independent factors, b0 is the intercept, bi the linear coefcients, bij the
quadratic coefcients, bij the cross-product coefcients, and k the
total number of independent factors. A backward regression procedure was used to determine the parameters of the models. This procedure systematically removed the effects that were not
signicantly associated (p > 0.05) with the response until a model
with only a signicant effect was obtained.
Table 2
F- and p-values of the ANOVA analysis for the quadratic model (Eq. 3) developed to
describe the inuence of electric eld strength (E) and treatment time (t) on the cell
disintegration index (Zp) of purple-eshed potato (PFP).
E
t
E2
t2
Model
Lack of t
r2
Adjusted-r2
Signal-to-noise ratio
Variation coefcient
p < 0.05 is signicant.
F-value
p-Value
103.80
137.08
9.71
20.72
65.80
0.66
0.98
0.97
22.55
16.25
<0.0001
<0.0001
0.0067
0.0003
<0.0001
>0.05
1333
105.00
90.00
0.9
0.8
75.00
0.7
0.6
60.00
0.5
0.4
0.3
45.00
2.00
2.50
3.00
3.50
4.00
Table 3
Anthocyanin extraction yield (AEY) (mg/100 g fw) resulting from the experimental conditions investigated for the untreated purple-eshed potato (untreated PFP) and PEF
treated purple-eshed potato (PEF-treated PFP) (3.4 kV/cm; 105 ls).
Extraction time (min)
60
60
60
60
60
270
270
270
270
270
480
480
480
480
480
Temperature (C)
10
10
25
40
40
10
25
25
25
40
10
10
25
40
40
Ethanol (%)
0
96
48
0
96
48
0
48
96
48
0
96
48
0
96
AEY untreated
a
8.1 1.04
15.2 0.28a
19.2 0.78a
25.2 0.35a
30.6 0.65a
29.8 0.39a
36.8 1.,53a
39.1 1.55a
49.2 2.68a
52.5 2.23a
28.9 1.22a
41.9 0.90a
50.5 0.14a
61.5 1.44a
63.9 1.68a
AEY PEF-treated
14.1 2.23b
17.1 0.05b
25.1 1.87b
29.9 0.38b
30.7 2.42a
35.9 0.77b
46.8 0.74b
50.5 0.86b
51.3 2.43a
55.6 1.85a
41.8 0.73b
49.5 3.06b
62.5 0.85b
65.8 1.11b
67.9 2.62a
t
T
Ec
t2
T2
tT
Model
r2
Adjusted-r2
Signal-to-noise ratio
Variation coefcient
p < 0.05 is signicant.
Untreated PFP
PEF-treated PFP
F-value
p-Value
F-value
p-Value
364.16
199.35
26.86
26.85
10.10
125.46
0.99
0.98
38.38
6.68
<0.0001
<0.0001
0.0006
0.0006
0.0112
<0.0001
990.63
284.97
11.15
33.98
15.41
7.19
229.09
0.99
0.99
47.86
3.99
<0.0001
<0.0001
0.0102
0.0004
0.0044
0.0279
<0.0001
The main difference between Eqs. (4 and 5) was that the square
of temperature was also a signicant term for anthocyanin extraction in PEF-treated PFP. The negative effect of this square term
indicates that in the range of temperatures investigated when
the PFP is treated by PEF above a maximum value, the increment
of temperature will not signicantly increase the extraction yield.
It has been estimated that the average diffusion coefcient of a
small solute in a membrane is often about a million times lower
than that in the adjacent aqueous solutions (Nobel, 1999). Therefore, the inuence of the temperature observed in the PEF-treated
PFP could be a consequence of the fact that the effect of the temperatures in anthocyanin diffusivity is lower when the cell membranes are permeabilised by the application of the PEF treatment.
In order to illustrate the advantages of the PEF permeabilisation
of the PFP before extraction in terms of increasing AEY or reducing
temperature and concentration of ethanol, Fig. 2 was plotted using
the corresponding regression models for untreated and PEF-treated
PFP (Eqs. 4 and 5). Fig. 2 shows that AEY increased with treatment
temperature and ethanol concentration in both untreated and PEFtreated samples. Independently of the extraction temperature, the
use of ethanol as solvent was more effective for the untreated PFP
than for the PEF-treated PFP. For example, for the untreated PFP at
25 C, the AEY using 96% ethanol was 20% higher than using water
as solvent, but only 5% for the PEF-treated PFP. This decrease of the
efcacy of ethanol as solvent as compared with water when samples were treated by PEF could be due to the high solubility of
anthocyanins in ethanol and the fact that the improvement of
extraction due to the increment of the diffusivity as a consequence
of the ethanolic denaturation of the phospholipid bilayer is less
signicant when the cells have been previously permeabilised by
PEF (Lapornik, Prosek, & Wondra, 2005).
A possible benet of the PEF permeabilisation of the PFP before
extraction is the possibility of decreasing the extraction temperature without affecting the AEY. For example, for a constant AEY
of 60 mg/100 g fw, the application of a PEF treatment permitted
the reduction of the extraction temperature from 40 to 25 C using
water as solvent and from 31 to 20 C when the solvent was 96%
80
70
1334
60
50
40
30
20
10
15
20
25
30
35
40
Temperature (C)
Fig. 2. Inuence of extraction temperature (C) and ethanol concentration (0%, solid
line; 48%, segmented line; 96%, dotted line) on anthocyanin extraction yield (AEY;
mg/100 g fw) for both untreated purple-eshed potato (untreated PFP; red colour)
and PEF-treated purple-eshed potato (PEF-treated PFP; green colour) (3.4 kV/cm;
105 ls) after an extraction time of 480 min. (For interpretation of the references to
colour in this gure legend, the reader is referred to the web version of this article.)
1335
Fig. 3. HPLC chromatograms of anthocyanin proles of aqueous extract of purple-eshed potato (PFP) at 520 nm for (A) control and (B) PEF treated sample (3.5 kV/cm102 ls). 1, malvidin 3-O-rutinoside-5-O-glucoside; 2, petunidin 3-O-caffeoyl-rutinoside-5-O-glucoside; 3, delphinidin 3-O-p-coumaroyl-rutinoside-5-O-glucoside; 4,
petunidin 3-O-p-coumaroyl-rutinoside-5-O-glucoside; 5, petunidin 3-O-feruloyl-rutinoside-5-O-glucoside; 6, malvidin 3-O-p-coumaroyl-rutinoside-5-O-glucoside; 7,
malvidin 3-O-feruloyl-rutinoside-5-O-glucoside.
ethanol. The potential of reducing extraction temperature by permeabilisation of cells by PEF has been observed by other authors
in the extraction of other compounds such as sugar from sugar
beets (Loginova, Vorobiev, Bals, & Lebovka, 2011; Lpez, Purtolas,
Condn, Raso, & lvarez, 2009). Finally, it is observed that in the
range of extraction temperatures between 35 and 40 C, where
the AEY was the highest for all solvents, the quantity of anthocyanins extracted from the untreated sample using 96% ethanol as solvent was similar to those extracted in the PEF-treated PFP using
water or ethanol. Therefore, in this range of temperatures, the
application of a PEF treatment to the PFP before extraction could
permit the use of water, a more environmentally friendly solvent
than ethanol, without decreasing the amount of anthocyanins
recovered from PEP.
3.4. HPLC characterisation of anthocyanins extracted from purpleeshed potato treated by PEF
Reverse-phase HPLC chromatogram proles detected at 520 nm
for untreated and PEF-treated PFP are presented in Fig. 3A and B.
HPLC chromatograms were similar to those obtained by Hillebrand
et al. (2011) for the same PFP cultivar. The PFP cultivar Vitelotte
contains ve minor anthocyanins and two major anthocyanins that
correspond to petunidin 3-p-coumaroylrutinoside-5-glucoside and
malvidin 3-p-coumaroyl-rutinoside-5-glucoside. The results of the
HPLC analyses showed that the application of a PEF treatment to
the PFP before extraction did not affect the extraction of a determined anthocyanin. The only difference observed was that the
peak areas were approximately 20% higher in the chromatograms
corresponding to the PEF-treated PFP. Similar results were obtained by Lpez, Purtolas, Hernndez-Orte, lvarez, and Raso
(2009) when comparing the anthocyanin prole of a control wine
with a wine obtained from grapes treated by PEF.
4. Conclusions
In this investigation, it has been demonstrated that independently of the extraction temperature or solvent used for extraction
of anthocyanins from PFP, the extraction yield was always higher
References
Angersbach, A., Heinz, V., & Knorr, D. (1999). Electrophysiological model of intact
and processed plant tissues: Cell disintegration criteria. Biotechnology Progress,
15, 753762.
Bridgers, E. N., Chinn, M. S., & Truong, V. D. (2010). Extraction of anthocyanins from
industrial purple-eshed sweet potatoes and enzymatic hydrolysis of residues
for fermentable sugars. Industrial Crops and Products, 32, 613620.
Brigita, L., Mirko, P., & Alenka, G. W. (2005). Comparison of extracts prepared from
plant by-products using different solvents and extraction time. Journal of Food
Engineering, 71, 214222.
Brown, C. R., Culley, D., Yang, C. P., Durst, R., & Wrolstad, R. (2005). Variation of
anthocyanin and carotenoid contents and associated antioxidant values in
potato breeding lines. Journal of the American Society for Horticultural Science,
130, 174180.
Cacace, J. E., & Mazza, G. (2003). Optimization of extraction of anthocyanins from
black currants with aqueous ethanol. Journal of Food Science, 68, 240248.
Cevallos-Casals, B. A., & Cisneros-Zevallos, L. (2003). Stoichiometric and kinetic
studies of phenolic antioxidants from Andean purple corn and red-eshed
sweet potato. Journal of Agricultural and Food Chemistry, 51, 33133319.
Chalermchat, Y., Fincan, M., & Djmek, P. (2004). Pulsed electric eld treatment for
solidliquid extraction of red beetroot pigment: Mathematical modelling of
mass transfer. Journal of Food Engineering, 64, 229236.
Corrales, M., Toep, S., Butz, P., Knorr, D., & Tauscher, B. (2008). Extraction of
anthocyanins from grape by-products assisted by ultrasonics, high hydrostatic
pressure or pulsed electric elds: A comparison. Innovative Food Science and
Emerging Technologies, 9, 8591.
De Vito, F., Ferrari, G., Lebovka, N. I., Shynkaryk, N. V., & Vorobiev, E. (2008). Pulse
duration and efciency of soft cellular tissue disintegration by pulsed electric
elds. Food and Bioprocess Technology, 1, 307313.
Donsi, F., Ferrari, G., & Pataro, G. (2010). Application of pulsed electric elds
treatments for the enhancement of mass transfer from vegetable tissue. Food
Engineering Reviews, 2, 109130.
El-Belghiti, K., & Vorobiev, E. (2005). Modelling of solute aqueous extraction from
carrots subjected to a pulsed electric eld pre-treatment. Biosystems
Engineering, 90, 289294.
Fan, G., Han, Y., Gu, Z., & Chen, D. (2008). Optimizing conditions for anthocyanins
extraction from purple sweet potato using response surface methodology
(RSM). LWT Food Science and Technology, 41, 155160.
1336
Fincan, M., DeVito, F., & Dejmek, P. (2004). Pulsed electric eld treatment for solid
liquid extraction of red beetroot pigment. Journal of Food Engineering, 64,
381388.
Francis, F. J. (1989). Food colorants: Anthocyanins. Critical Reviews in Food Science
and Nutrition, 28, 273314.
Gaschovska, T., Cassada, D., Subbiah, J., Hanna, M., Thippareddi, H., & Snow, D.
(2010). Enhanced anthocyanin extraction from red cabbage using pulsed
electric eld processing. Journal of Food Science, 75, 323329.
Giusti, M. M., & Wrolstad, R. E. (2001). Unit F1.2. Anthocyanins. Characterization
and measurement with UV visible spectroscopy. In R. E. Wrolstad (Ed.), Current
protocols in food analytical chemistry (pp. 113). New York: Wiley.
Goldstein, D. B., & Chin, J. H. (1981). Interaction of ethanol with biological
membranes. Federation Proceedings, 40, 20732076.
He, J., & Giusti, M. M. (2010). Anthocyanins: Natural colorants with healthpromoting properties. Annual Review of Food Science and Technology, 1, 163187.
Hillebrand, S., Husing, B., Schliephake, U., Trautz, D., Herrmann, M. E., &
Winterhalter, P. (2011). Effect of thermal processing on the content of
phenols in pigmented potatoes (Solanum tuberosum L.). Enrahrungs Umschau,
58, 349353.
Hojnik, M., Skerget, M., & Knez, Z. (2008). Extraction of lutein from marigold ower
petals Experimental kinetics and modeling. LWT Food Science and
Technology, 40, 20082016.
Ieri, F., Innocenti, M., Andrenelli, L., Vecchio, V., & Mulinacci, N. (2011). Rapid HPLC/
DAD/MS method to determine phenolic acids, glycoalkaloids and anthocyanins
in pigmented potatoes (Solanum tuberosum L.) and correlations with variety and
geographical origin. Food Chemistry, 125, 750759.
Jansen, G., & Flamme, W. (2006). Coloured potatoes (Solanum tuberosum L.)
Anthocyanin content and tuber quality. Genetic Resources and Crop Evolution, 53,
13211331.
Knorr, D., Froehling, A., Jaeger, H., Reineke, K., Schlueter, O., & Schoessler, K. (2011).
Emerging technologies in food processing. Annual Review of Food Science and
Technology, 2, 203235.
Lachman, J., Hamouz, K., Sulc, M., Orsk, M., Pivec, V., Hejtmnkov, A., et al. (2009).
Cultivar differences of total anthocyanins and anthocyanidins in red and
purple-eshed potatoes and their relation to antioxidant activity. Food
Chemistry, 114, 836843.
Lachman, J., Hamouz, K., Sulc, M., Orsk, M., Pivec, V., Hejtmnkov, A., et al. (2012).
Impact of selected factors Cultivar, storage, cooking and baking on the content
of anthocyanins in coloured-esh potatoes. Food Chemistry, 133, 11071116.
Lapornik, B., Prosek, M., & Wondra, A. G. (2005). Comparison of extracts prepared
from plant by-products using different solvents and extraction time. Journal of
Food Engineering, 71, 214222.