Journal of Food Processing and Preservation ISSN 1745-4549

PROCESSING OF RED WINE BY PULSED ELECTRIC FIELDS WITH

RESPECT TO QUALITY PARAMETERS
ELIF E. ABCA1 and GULSUN AKDEMIR EVRENDILEK2,3
1
Ministry of Food Agriculture and Livestock, Ankara, Turkey
2
Faculty of Engineering and Architecture, Department of Food Engineering, Abant Izzet Baysal University, Bolu 14280, Turkey

3
Corresponding author. ABSTRACT
TEL: +90-374-253-2600 ext. 2619;
FAX: +90-374-253-45-58; Processing of red wine by pulsed electric fields (PEFs) with determination of
EMAIL: gevrendilek@yahoo.com; physical, chemical and sensory properties as well as inactivation of Escherichia coli
gevrendilek@ibu.edu.tr O157:H7, Lactobacillus delbrueckii ssp. bulgaricus, Candida lipolytica, Saccharomy-
ces cerevisiae and Hansenula anomala revealed that increased electric field strength
Received for Publication April 2, 2014
Accepted for Publication April 29, 2014
and treatment temperature did not cause significant difference on measured attri-
butes (P > 0.05); however, initial number of inoculated microorganisms signifi-
doi:10.1111/jfpp.12285 cantly decreased (P ≤ 0.05). Response surface methodology was used to design the
experiments, and a central composite design with quadratic model fitted to
explain microbial inactivation. It is concluded that processing of red wine with no
significant changes and microbial stability can be ensured with PEF treatment.

PRACTICAL APPLICATIONS
It was revealed that sulfur dioxide used in vineries to stop the fermentation and
eliminate growth of wild yeasts may cause allergic reactions, asthma and head-
ache, and thus it is recommended that sulfur dioxide dose needs to be decreased.
Alternative methods such as pulsed electric fields (PEFs) are in search to examine
the potential applications in wine-making industry. It was shown by this study
that PEF has a potential to process red wine without adversely affecting important
properties.

(Martin-Belloso et al. 1997; Evrendilek et al. 1999; Cortés

INTRODUCTION
Recent demands from consumers for fresh like quality foods
with less and/or no artificial additives have introduced
innovative processing technologies as an alternative to the
traditional ones that may lead to improve the competitive-
ness of the food industry by upgrading food quality, intro-
ducing new products in the market and reducing the cost of
energy (Tiwari et al. 2009). Pulsed electric field (PEF) is one
of the most promising novel processing techniques that
have drawn considerable attention owing to their involve-
ment of minimal food processing with fewer preservatives.
PEFs involve the application of very short-duration
pulses of high electric field strength at the magnitude of
20–80 kV/cm to food material located between two elec-
trodes (Sale and Hamilton 1967; Zimmerman et al. 1974;
Knorr and Angersbach 1998). PEF was successfully applied
to different food products including fruit and vegetable
juices, milk and milk products, soup, and beer
et al. 2005; Rivas et al. 2006; Jin and Zhang 2007).
Wine is one of the most consumed and most favored bev-
erages worldwide made of fermented fruit juice, usually
from grapes (Ancín et al. 2004; Arapitsas et al. 2004;
Garde-Cerdán et al. 2004; Morales et al. 2004). During and
after fermentation, wine can be spoiled by different micro-
organisms that are part of the natural microbiota of grape
skins and wine contact surfaces such as winery equipment
and barrels (Couto et al. 2005). Addition of sulfur dioxide
(SO2) is a usual practice in wineries to decrease the risk of
microbial spoilage during the wine-making process and fol-
lowing the conclusion of the fermentation even though the
resistance and/or sensitivity of different strains of microor-
ganisms to SO2 vary considerably (Du Toit and Pretorius
2000). In addition, due to recent findings related to adverse
effects of SO2 such as allergy, asthma and headache, the rec-
ommended dose of SO2 decreased (Usseglio-Thomasset
1992). Thus, alternative methods such as PEFs are in search

Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc. 1
PEF PROCESSING OF RED WINE
to examine the potential applications in the wine-making
industry. PEF technology has mainly been tested for the
improvement of the extraction of phenolic compounds
during the maceration process, acceleration of wine aging
and inactivation of spoilage bacteria (Garde-Cerdán et al.
2007; Lopez et al. 2008, 2009; Puertolas et al. 2009, 2010a).
However, more studies are needed to determine the effect of
PEF on wine quality, sensory properties and microbial inac-
tivation. Therefore, this study was planned to determine the
effect of PEF in combination with moderate heat treatment
on physicochemical and sensory properties of red wine as
well as the inactivation of yeasts and bacteria.
MATERIALS AND METHOD
Wine Samples
The red wine samples with 12% alcohol content from
Bogazkere and Okuzgozu grapes in Elazig Province of
Turkey before the ending fermentation were obtained from
Dimes Gida San ve Tic A.S. (Tokat, Turkey), and treated by
PEF immediately after receiving. The samples were cooled
down to room temperature immediately and taken for
analyses.
Test Microorganisms
The cultures of Saccharomyces cerevisiae, Hansenula
anomala, Candida lipolytica (Yarrowia lipolytica) and Lacto-
bacillus delbrueckii ssp. bulgaricus were obtained from
Ankara University Food Engineering Department culture
collection (Ankara, Turkey) in tryptic soy broth (TSB)
(Fluka, Munich, Germany). In order to activate, the yeasts
were transferred into TSB and incubated at 22 ± 2C for
12 h, and this procedure was repeated at least three times.
After that, they were inoculated in wine samples separately.
The culture of L. delbrueckii ssp. bulgaricus was obtained in
De Man Rogosa Sharpe (MRS) broth (Fluka) and inocu-
lated into wine samples after transferring the culture to
MRS broth and following incubation at 35 ± 2C for 12 h.
Escherichia coli O157:H7 (EDL 931 04054) culture was
obtained from Refik Saydam Hıfzıssıhha Research Center
Culture Collection Laboratory (Ankara, Turkey) in lyophi-
lized form. The culture was activated with three consecutive
transfers into McConkey Sorbitol Agar (Fluka) and follow-
ing incubation at 35 ± 2C. After activation, the culture was
inoculated into wine samples.
PEF Processing
OSU-4A bench scale continuous PEF system purchased
from the Ohio State University (Columbus, OH) providing
2 Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
E.E. ABCA and G. AKDEMIR EVRENDILEK
square wave bipolar pulses was used. The system equipped
with six treatment chambers having 0.29 cm in diameter
and 0.23 cm gap distance. The post- and pretreatment tem-
peratures (t2–t1, t4–t3 and t6–t5) at the outlet and inlet of each
pair of treatment chambers were monitored by K-type dual-
channel digital thermocouples (Fisher Scientific, Pittsburgh,
PA) attached to the outer surface of the thin-wall stainless
steel tubing. The treated sample was cooled after each pair
of chambers by cooling coils submerged in a water bath at
12C (model RTE-111; NESLAB Instruments, Inc., Newing-
ton, NH). A trigger generator (model 9300 series; Quantum
Composers, Inc., Bozeman, MT) was used to control pulse
duration time, pulse delaying time and pulse repetition rate.
Applied voltage and current were monitored by a two-
channel digital oscilloscope (model TDS 320; Tektronix,
Inc., Beaverton, OR). Specifications of the OSU-4A bench
scale PEF units were 12,000 V max of output voltage, 60 A
max of output current, 10.000 pulse per second (pps) max
of repetition rate, 200–1200 Ω of load resistance and 16 J of
energy storage in the pulse generator when fully charged.
Preliminary tests were performed to determine PEF treat-
ment conditions for red wine samples. For processing of
wine samples, different electric field strengths of 0 (control),
17, 24 and 31 kV/cm at 10, 20 and 30C were applied. For all
treatments 40 mL/min of flow rate, 3 μs of pulse duration
and 500 pps of frequency were used.
Physicochemical Analysis of Red Wine
pH measurement was conducted by 10 mL of the triplicate
samples at room temperature by Orion perpHect logR
meter (inoLab WTW, Weilheim, Germany). Titratable
acidity (TA) measurement was calculated with reference to
tartaric acid (g/L). Conductivity of the samples was mea-
sured by handheld conductivity meter (sensION 5 model,
HACH, Loveland, CO), and the results were given as
mS/cm. Color measurement was performed by Hunter
color flex spectrophotometer (Hunter Associates Labora-
tory, Inc., Reston, VA) using CIELAB color scale at D65/10°
to measure L, a and b parameters. The hue (arctan b/a) and
chroma ( a2 + b 2 ) of the wine samples were also calculated.
In order to measure the total monomeric anthocyanin
(TMAC) content of red wine, the samples were diluted with
0.025 M KCl (Sigma Chemical, Co., Stockholm, Sweden)
and 0.04 M sodium acetate (Sigma Chemical Co.), sepa-
rately, and the mixtures were centrifuged at 2,400 rpm for
2 min by vortex. The samples were set for 20 min, and both
dilutions were read at both 520 and 700 nm using a spectro-
photometer (Lambda 25 model, Perkin Elmer, Waltham,
MA). Results were calculated as cyanidin 3-glucoside
equivalent in mg/L (Lee et al. 2005).
For the total antioxidant capacity (TAC) analysis, Tris-
HCl tampon at pH 7.4 was added to 0.1 mL of the samples.
E.E. ABCA and G. AKDEMIR EVRENDILEK
The mixture was vortexed at 2,400 rpm for 5 min, and 1 mL
of DPPH (prepared in ethanol) was added. The absorbance
of red wine samples was measured at 517 nm wavelength.
Total antioxidant activity (%) was calculated according to
Moon and Terao (1998).
Total phenolic substance content (TPSC) of the samples
was measured by the spectrophotometric method devel-
oped by Spanos and Wrolstad (1990). Obtained absorbance
values were calculated from the gallic acid standard curve
prepared with 100, 200, 300, 400 and 500 mg/L gallic acid.
The total phenolic content of the samples was calculated as
mg/L gallic acid equivalence.
The metal ion concentration of the samples was mea-
sured with inductively coupled plasma mass spectroscopy
(ICP-MS) (XSERIES 2, Thermo Scientific, Schwerte,
Germany). One milliliter of the sample was taken and
0.5 mL of hydrogen peroxide was added before adding
2.5 mL of 65 % nitric acid. After complete digestion of the
samples (30 min) at room temperature, they were held for
20 min at 140C in microwave (CEM MARS 5, ABD). The
samples were then filtered through 0.45 μm filter hydro-
philic PVDF (Millipore Millex-Merck Chemicals GmbH,
Schwalbach, Germany), and the filtered samples were com-
pleted to 10 mL with distilled water. Standard metal solu-
tion was prepared daily from 1,000 mg/L stock (Merck,
Darmstadt, Germany) in 2% nitric acid Suprapur grade
(Merck). ICP-MS had babington-type nebulizer and scott-
type spray chamber, RF generator that has 10 MHz fre-
quency and 1,300 power output. Argon flow rate (L/min)
was 15 for plasma, 0.9 for auxiliary and 1–1.1 for nebulizer.
Solution uptake rate was 1.8 mL/min. Sampler cone and
skimmer were nickel with i.d. of 1.1 and 0.9 mm, respec-
tively. Pressure of interface and quadruple were 4 and
2 × 10−6 torr. Data acquisition was provided by peak
hopping, 200 ms replicate time, 200 ms dwell time, 3 sweeps
per reading and 3 readings per replicates. Analytical masses
were 75As, 40Ca, 24Mg, 111Cd, 63Cu, 52Cr, 56Fe, 202Hg, 39K, 23Na,
24
Mg, 55Mn, 31P, 208Pb, 80Se, 118Sn and 66Zn (Cubadda et al.
2001).
Microbial Enumeration
Both control and PEF-treated samples inoculated with bac-
teria and yeasts separately were diluted with 0.1% peptone
(Fluka) water, and 100 μL of appropriate dilutions was
plated into MacConkey Sorbitol Agar (Fluka) for E. coli
O157:H7, yeast extract peptone glucose (Fluka) for
S. cerevisiae, H. anomala and C. lipolytica, and MRS agar
(Fluka) for L. delbrueckii ssp. bulgaricus by surface plating
method separately. The plates for E. coli O157:H7 and
L. delbrueckii ssp. bulgaricus were incubated at 35 ± 2C for
24–48 h and plates for S. cerevisiae, H. anomala and
C. lipolytica were incubated at 22 ± 2C for 3–5 days. Plates
Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
PEF PROCESSING OF RED WINE
for L. delbrueckii ssp. bulgaricus were incubated under
anaerobic conditions using anaerobic jar. Results were cal-
culated as log colony-forming unit (cfu)/mL.
Sensory Analyses
Red wine samples at the temperature of 15–18C were ana-
lyzed for sensory properties involving visual evaluation,
smell and taste by 50 trained panelists. For visual evalua-
tion, they were asked to evaluate the samples for being
cloudy or clear (cloudiness/clearness), dull or bright
(dullness/brightness), red color intensity, density and par-
ticle status by tilting and holding the glass in front of the
white paper given. For smell, they were asked to swirl the
glasses for 0–12 s, take a quick whiff to gain a first impres-
sion and take a deep breath to receive the whole impression.
This was repeated again to complete smell evaluation for
odor/flavor. Finally, they were asked to taste the samples.
For this step they were asked to take a sip, roll it all around
the mouth and evaluate the samples in three stages: the
initial attack phase, the evaluation phase and finish to evalu-
ate the samples for wine taste, bitter taste and sour taste.
Finally, they were asked to swallow the samples for the
evaluation of aftertaste. They were informed that they must
drink water between the samples, consume couple bits of
unsalted cracker to clean their plate and drink water again
before the evaluation of the next sample. The sensory panel
was conducted based on a 9-point hedonic scale. In order to
reduce the number of samples, only 0 kV/cm at both 10 and
30C and 17, 24 and 31 kV/cm at 30C samples were sub-
jected to sensory analyses.
Data Analyses
Response surface methodology was used to determine the
influence of temperature (x1) and electric field strength (x2)
on microbial inactivation (number of surviving cells after
PEF processing at three levels). Each variable was coded are
three levels (−1, 0, +1) according to the following equation.
( xi − xi )
Xi = (1)
Δxi
where Xi is the dimensionless value of an independent vari-
able, xi is the real value of an independent variable, xi is the
real value of an independent variable at center point and Δxi
is the step change. A central composite design (CCD) was
arranged to allow for fitting of quadratic model (Nakai et al.
2006). The CCD combined the vertices of hybercube whose
coordinates were given by 2n factorial design with three rep-
licates (∝ = 1.41) at the center point. The design of the
experiments is presented in Table 1. All experiments were
carried out in order to minimize any effect of extraneous
3
PEF PROCESSING OF RED WINE E.E. ABCA and G. AKDEMIR EVRENDILEK

TABLE 1. EXPERIMENTAL DESIGN

Variables Responses (survival log cfu/mL)

Escherichia coli Candida Lactobacillus Saccharomyces

Runs X1 X2 O157:H7 lipolytica H. anomala bulgaricus cerevisiae
1 1.00 1.00 4.96 5.30 5.29 6.13 4.38
2 1.00 −1.00 4.60 5.17 5.08 6.14 5.95
3 −1.00 1.00 4.30 3.30 2.54 1.94 1.13
4 0.00 0.00 1.26 3.90 2.40 6.85 4.00
5 0.00 −1.41 4.54 2.41 5.14 5.59 5.77
6 0.00 0.00 4.47 2.14 0.00 3.45 4.03
7 0.00 1.41 3.58 4.30 3.00 4.00 0.00
8 1.00 −1.00 3.55 2.14 2.54 3.60 5.95
9 0.00 0.00 3.46 2.24 0.00 3.68 3.93
10 0.00 1.41 2.82 2.30 3.00 4.02 0.00
11 0.00 0.00 2.20 2.41 0.00 3.99 3.93
12 0.00 −1.41 2.37 2.01 2.08 3.04 5.97
13 1.41 0.00 4.90 3.90 1.70 2.47 3.19
14 0.00 0.00 4.98 3.55 0.00 2.14 4.03
15 −1.00 1.00 4.92 2.17 1.23 5.50 1.43
16 1.41 0.00 4.55 1.30 5.73 5.51 3.07
17 −1.00 1.00 4.60 2.14 0.72 1.94 1.13
18 0.00 0.00 3.89 2.94 2.08 3.39 3.83
19 −1.41 0.00 3.26 0.30 5.35 5.14 3.30
20 0.00 0.00 3.25 3.64 2.08 5.50 4.03
21 −1.00 −1.00 3.15 0.00 0.00 6.13 5.70
22 0.00 0.00 2.04 0.00 1.14 4.00 4.03
23 0.00 0.00 2.01 2.42 5.47 2.04 4.38
24 1.00 1.00 2.73 2.41 2.09 6.80 0.00
25 −1.41 0.00 4.96 2.14 4.76 3.45 3.00
26 0.00 1.41 4.98 2.41 1.23 2.68 0.00
27 −1.41 0.00 4.80 0.90 3.00 4.00 2.12
28 0.00 0.00 4.17 5.30 0.00 3.80 4.38
29 0.00 0.00 3.55 5.30 5.35 2.74 4.00
30 1.00 1.00 3.55 3.42 1.14 2.20 1.02
31 1.00 −1.00 2.62 3.55 0.47 5.23 5.95
32 0.00 0.00 2.61 2.14 0.00 3.80 3.91
33 −1.00 −1.00 2.55 2.17 3.00 6.85 5.70
34 0.00 0.00 1.14 0.00 5.47 3.80 3.93
35 1.41 0.00 1.01 1.47 2.10 3.80 3.30
36 −1.00 −1.00 1.00 2.24 5.14 5.72 5.70
37 1.00 1.00 1.09 1.40 2.92 3.77 0.00
38 0.00 −1.41 4.92 3.30 4.80 5.23 5.77
39 0.00 0.00 3.67 0.00 0.00 7.74 4.03

cfu, colony-forming unit.

factors on the observed responses. The model proposed for significant (P < 0.05) at 95% confidence level. The data
response (Y) was: related to physical properties of red wine samples were ana-
lyzed using one-way and/or two-way ANOVA at 95% confi-
2 2 2
dence interval. Differences between electric field strengths
Y = β0 + ∑ βii X i + ∑ βii X i2 + ∑ ∑β XX ii i j (2)
i =1 i =1 i ≺ j =1 were determined by Tukey’s multiple comparison test
(Minitab 15 version, Minitab, Inc., State College, PA).
where β0, βi, βii and βj are the regression coefficients for
intercept, linear, quadratic and interaction terms, respec-
tively, and Xi and Xj are the independent variables.
RESULTS AND DISCUSSIONS
The statistical significance of the second-order model
equation was evaluated by the F test analysis of variance Water bath temperature was adjusted to 10, 20 and 30C to
(ANOVA), which revealed that this regression is statistically determine the effect of increased temperature on the quality

4 Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
E.E. ABCA and G. AKDEMIR EVRENDILEK PEF PROCESSING OF RED WINE

of wine samples during PEF processing of wine samples.

TPSC (g/100 mL 0.0445 ± 0.005 0.0445 ± 0.005 0.0445 ± 0.005 0.0442 ± 0.002 0.0443 ± 0.003 0.0446 ± 0.004 0.0443 ± 0.003 0.0444 ± 0.003 0.0452 ± 0.002 0.0443 ± 0.004 0.0443 ± 0.003 0.0441 ± 0.003
2.94 ± 0.04
2.36 ± 0.01

4.90 ± 0.50

2.39 ± 0.37
15.56 ± 2.07
2.69 ± 0.04
5.72 ± 1.42
15.79 ± 2.01

59.45 ± 2.35
28.96 ± 3.44
With increased electric field strength from 0 to 31 kV/cm,
sample temperatures increased from 10 ± 2C to 40 ± 2C.

30C
After PEF treatment, the samples immediately were cooled
down to room temperature by immersing into ice bath

3.02 ± 0.01

5.40 ± 0.30

2.57 ± 1.41
14.67 ± 0.13
2.57 ± 0.75
5.65 ± 1.23
14.89 ± 0.89

59.10 ± 0.35
28.55 ± 2.43
2.31 ± 0.0
before analyses. Measured sulfur dioxide level was <20 mg/L
in red wine samples used in the study.

20C
Control samples had a pH of 3.01 ± 0.04, conductivity of

TABLE 2. MEASUREMENT OF PHYSICAL PROPERTIES OF RED WINE PROCESSED BY PEF AS A FUNCTION OF ELECTRIC FIELD STRENGTH AND TREATMENT TEMPERATURE
2.31 ± 0.01 mS/cm and TA of 5.00 ± 0.30 g/L tartaric acid at

3.07 ± 0.02
2.28 ± 0.01

5.40 ± 0.20

2.56 ± 0.11
12.35 ± 0.15
2.70 ± 1.28
7.20 ± 1.15
12.46 ± 1.22

59.83 ± 1.21
29.53 ± 2.34
10, 20 and 30C. With increased electric field strength and
treatment temperature, pH, conductivity and TA of the
samples were not significantly changed (P > 0.05) (Table 2).

10C
31
Regardless of the applied electric field strength and treat-

2.96 ± 0.04
2.36 ± 0.03

5.00 ± 0.40

3.15 ± 0.51
14.48 ± 2.06
2.70 ± 0.06
5.30 ± 1.06
14.72 ± 1.12

59.67 ± 0.29
29.32 ± 2.93
ment temperature, no significant difference was detected for
“L,” “a” and “b” values (P > 0.05) (Table 2). Hue and chroma

PEF, pulsed electric field; TA, titratable acidity; TAC, total antioxidant capacity; TMAC, total monomeric anthocyanin content; TPSC, total phenolic substance content.
30C
values also did not significantly change with increased elec-
tric field strength and treatment temperature (P > 0.05)

3.04 ± 0.02
2.31 ± 0.00

5.50 ± 0.70

2.70 ± 0.60
14.67 ± 2.18
2.88 ± 0.65
5.02 ± 1.42
14.95 ± 1.35

59.87 ± 0.70
29.88 ± 3.04
(Table 2).
TPSC of the control samples was 0.0445 ± 0.003 mg/mL

20C
gallic acid equivalence, and this value did not change
with increased temperature and/or electric field strength

3.08 ± 0.03
2.28 ± 0.02

5.50 ± 0.30

2.80 ± 0.13
12.40 ± 0.14
2.56 ± 0.14
4.76 ± 0.14
12.66 ± 1.14

60.10 ± 0.39
32.20 ± 2.09
(P > 0.05) (Table 2). TACs of the control samples were
60.69 ± 1.01, 59.83 ± 2.68 and 59.48 ± 2.89% at 10, 20 and
30C. There was a slight decrease on TAC with increased 10C
24

electric field strength and treatment temperature, but this

4.90 ± 0040
2.96 ± 0.03
2.35 ± 0.03

2.35 ± 0.26
13.20 ± 0.28
2.59 ± 0.20
5.01 ± 0.24
13.45 ± 1.24

60.83 ± 0.18
28.30 ± 2.63
difference was not significant (P > 0.05) (Table 2). It was
observed that with increased electric field strength and
treatment time, there was a slight decrease on TMAC of the
30C

samples; however, this difference was not significant among

3.04 ± 0.02
2.30 ± 0.00

5.70 ± 0.30

2.27 ± 0.60
14.35 ± 3.12
2.80 ± 0.75
5.05 ± 1.93
14.62 ± 1.75

60.44 ± 0.63
29.62 ± 2.53
the control and PEF-treated samples (P > 0.05) (Table 2).
The measured red wine samples revealed that the amount
of K was highest with 2,000.00 ± 86.13 ppm in control
20C

samples. Amounts of Mg, P, Ca, Na, Mn and Fe were

363.73 ± 45.38, 196.43 ± 17.03, 177.88 ± 19.73, 33.00 ± 9.72,
3.07 ± 0.03
2.30 ± 0.03

5.20 ± 0.10

2.33 ± 0.86
12.35 ± 1.09
2.70 ± 0.22
4.50 ± 0.15
12.64 ± 0.82

60.67 ± 0.37
33.73 ± 2.00
0.77 ± 1.25 and 7.19 ± 0.89 ppm, respectively. The amounts
of As, Cd, Cu, Hg, Pb, Se, Sn and Zn were very low, and
10C
17

amounts of Cd and Sn could not be detected. Application of

3.01 ± 0.03
2.31 ± 0.01

5.00 ± 0.30

2.17 ± 0.49
13.27 ± 2.84
2.64 ± 1.34
4.19 ± 2.09
13.53 ± 1.34

59.48 ± 2.89
28.22 ± 5.05

increased electric field strength and treatment time did not

cause significant difference on the measured concentrations
Data in the same row are not significantly different (P ≤ 0.05).

(P > 0.05) (Table 3).

30C

Initial number of E. coli O157:H7 in control samples was

4.97 ± 0.08 log cfu/mL. Application of 31 kV/cm at 10, 20
3.01 ± 0.03
2.31 ± 0.01

5.00 ± 0.30

2.17 ± 0.49
13.27 ± 2.84
2.64 ± 1.34
4.19 ± 2.09
13.53 ± 1.34

59.83 ± 2.68
30.12 ± 4.62
Electric field strength (kV/cm)

and 30C temperatures ended up with 2.73 ± 0.33,

2.37 ± 0.34 and 1.26 ± 0.12 log cfu/mL E. coli O157:H7
20C

counts, respectively (P ≤ 0.05) (Fig. 1; Table 4). Before PEF

application, initial number of L. delbrueckii ssp. bulgaricus
3.01 ± 0.03
2.31 ± 0.01

5.00 ± 0.30

2.17 ± 0.49
13.27 ± 2.84
2.64 ± 1.34
4.19 ± 2.09
13.53 ± 1.34

60.69 ± 1.01
TMAC (cyanidin 35.03 ± 4.18

inoculated in red wine was 6.14 ± 0.48 log cfu/mL. After

application of PEF processing at 10, 20 and 30C tempera-
10C

tures, the number of survivals was 3.39 ± 0.47, 2.74 ± 0.39,

0

2.68 ± 0.45 at 24 kV/cm and 3.45 ± 0.34, 2.47 ± 0.40 and

3-glucoside
equivalent)

equivalent
Conductivity

gallic acid

2.04 ± 0.19 at 31 kV/cm, respectively (P ≤ 0.05) (Fig. 1;

TA (tartaric
acid g/L)
(mS/cm)
properties

Color (b)
Color (a)
Color (L)

TAC (%)
Chroma

mg/L)
Physical

Table 4). Initial number of C. lipolytica was counted as

Hue

5.30 ± 0.24 log cfu/mL. After application of 31 kV/cm

pH

Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc. 5
PEF PROCESSING OF RED WINE E.E. ABCA and G. AKDEMIR EVRENDILEK

TABLE 3. MINERAL CONCENTRATION OF RED WINE SAMPLES PROCESSED BY PEF AS A FUNCTION OF ELECTRIC FIELD STRENGTH AND
TREATMENT TEMPERATURE

Electric field strength (kV/cm)

0 17 24 31
Mineral Temperature (C) Mineral concentration (ppm)
As 10 0.01 ± 0.00a 0.01 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a
20 0.01 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a
30 0.01 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a
Ca 10 177.88 ± 19.73a 176.90 ± 8.70a 179.68 ± 30.43a 172.09 ± 18.42a
20 177.88 ± 19.73a 176.26 ± 16.35a 174.36 ± 20.55a 172.44 ± 31.36a
30 177.88 ± 19.73a 155.38 ± 66.03a 177.85 ± 49.25a 196.90 ± 9.04a
Cu 10 0.09 ± 0.02a 0.03 ± 0.02a 0.03 ± 0.02a 0.04 ± 0.02a
20 0.09 ± 0.02a 0.03 ± 0.01a 0.04 ± 0.03a 0.03 ± 0.02a
30 0.09 ± 0.02a 0.09 ± 0.08a 0.05 ± 0.04a 0.05 ± 0.04a
Na 10 33.00 ± 9.72a 33.08 ± 2.44a 28.51 ± 6.39a 32.04 ± 8.54a
20 33.00 ± 9.72a 30.56 ± 7.26a 32.36 ± 9.50a 34.40 ± 7.36a
30 33.00 ± 9.72a 21.07 ± 28.67a 32.57 ± 21.05a 30.76 ± 8.18a
Mg 10 363.73 ± 45.38a 364.15 ± 9.75a 354.80 ± 36.70a 356.80 ± 42.50a
20 363.73 ± 45.38a 348.80 ± 36.50a 360.15 ± 49.25a 374.50 ± 70.60a
30 363.73 ± 45.38a 362.90 ± 35.10a 361.80 ± 26.70a 366.90 ± 21.90a
P 10 196.43 ± 17.03a 193.80 ± 8.46a 193.90 ± 8.02a 195.80 ± 7.45a
20 196.43 ± 17.03a 195.60 ± 10.00a 194.10 ± 9.88a 196.60 ± 11.04a
30 196.43 ± 17.03a 203.80 ± 7.93a 198.70 ± 6.90a 196.90 ± 9.04a
K 10 2,000.00 ± 86.13a 2,039.50 ± 88.50a 2,120.00 ± 85.50a 2,119.00 ± 88.95a
20 2,000.00 ± 8.13a 2,161.00 ± 615.85a 2,150.00 ± 76.90a 2,141.00 ± 84.05a
30 2,000.00 ± 8.13a 1,990.00 ± 1,085.05a 1,971.00 ± 80.15a 2,000.00 ± 75.65a
Fe 10 7.19 ± 0.89a 7.69 ± 0.23a 7.19 ± 0.85a 7.20 ± 0.09a
20 7.19 ± 0.89a 7.39 ± 0.51a 7.05 ± 0.77a 7.44 ± 0.71a
30 7.19 ± 0.89a 7.13 ± 2.23a 7.94 ± 1.63a 7.85 ± 1.52a
Hg 10 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a
20 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a
30 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a 0.04 ± 0.00a
Mn 10 10.77 ± 1.25a 8.46 ± 2.30a 9.26 ± 0.68a 9.65 ± 0.84a
20 10.77 ± 1.25a 9.43 ± 1.68a 9.68 ± 1.19a 9.07 ± 1.81a
30 10.77 ± 1.25a 8.38 ± 3.98a 10.01 ± 2.78a 10.87 ± 2.70a
Se 10 0.02 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.00a 0.01 ± 0.00a
20 0.02 ± 0.00a 0.01 ± 0.00a 0.03 ± 0.02a 0.03 ± 0.02a
30 0.02 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.00a
Pb 10 0.03 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.01a 0.02 ± 0.00a
20 0.03 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.00a 0.02 ± 0.00a
30 0.03 ± 0.00a 0.02 ± 0.01a 0.02 ± 0.01a 0.02 ± 0.01a
Zn 10 0.34 ± 0.04a 0.27 ± 0.02a 0.18 ± 0.04a 0.20 ± 0.04a
20 0.34 ± 0.04a 0.18 ± 0.03a 0.23 ± 0.02a 0.22 ± 0.02a
30 0.34 ± 0.04a 0.35 ± 0.18a 0.28 ± 0.14a 0.21 ± 0.09a

Data at the same row with different superscript letters are significantly different (P ≤ 0.05) (n = 6).
PEF, pulsed electric field.

electric field strength at 10, 20 and 30C, the number
C. lipolytica was counted as 0.90 ± 0.00, 0.00 ± 0.00 and
0.00 ± 0.00 log cfu/mL (P ≤ 0.05) (Fig. 1; Table 4). The
number of H. anomala in control samples was 5.29 ± 0.10
log cfu/mL. Processing of red wine samples at 30C with 17,
24 and 31 kV/cm electric field strengths reduced the initial
H. anomala number to 1.70 ± 0.23, 0.72 ± 0.09 and 0.00 ±
0.00 log cfu/mL, respectively (P ≤ 0.05) (Fig. 1; Table 4).
The number of S. cerevisiae in control samples was
5.95 ± 0.16 log cfu/mL. When electric field strength was
applied at 31 kV/cm, the number of S. cerevisiae was
reduced to 1.43 ± 0.27, 1.02 ± 0.07 and 0.00 ± 0.00 for 10, 20
and 30C (P ≤ 0.05) (Fig. 1; Table 4). Increased electric field
strength and treatment temperature caused a complete
inactivation of C. lipolytica, H. anomala and S. cerevisiae.
Inactivation of the all microorganisms inoculated into wine
samples was significant (P ≤ 0.05) (Fig. 1; Table 4). Inactiva-
tion rate of the microorganisms was H. anomala =
C. lipolytica = S. cerevisiae > E. coli O157:H7 > L. delbrueckii
ssp. bulgaricus.

6 Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
E.E. ABCA and G. AKDEMIR EVRENDILEK PEF PROCESSING OF RED WINE

A B

5 6
4
E. coli O157:H7 4
3 C. lipolitica

30 2
2 30
20 0
0 temperature (C) 20 temperature (C)
10 10 0
20 10
kV/cm 30 20 10
kV/cm 30

C D

6
6
4
P. anomola L. bulgaricus
2 4
30 30
0 20 2
temperature (C) 20
0 0
temperature (C)
10 10 10
20 20 10
kV/cm 30 kV/cm 30

4
S. cerevisiae

2
30
0
20 temperature (C)
0
10 10
20
kV/cm 30

FIG. 1. INACTIVATION OF ESCHERICHIA COLI O157:H7 (A), CANDIDA LIPOLYTICA (B), H. ANOMALA (C), LACTOBACILLUS DELBRUECKII SSP.
BULGARICUS (D) AND SACCHAROMYCES CEREVISIAE (E) BY PULSED ELECTRIC FIELD AS A FUNCTION OF ELECTRIC FIELD STRENGTH AND
TREATMENT TEMPERATURE

It was revealed that all microbial inactivation data fitted
response surface quadratic models. The predicted values of
microbial inactivation were calculated using the regression
model and compared with experimental values. The total
determination coefficients were changed among microor-
ganisms, and the R2 values for the inactivation of E. coli
O157:H7, C. lipolytica, H. anomala, L. delbrueckii ssp.
bulgaricus and S. cerevisiae for SRM validation were 91.2,
91.6, 91.8, 92.0 and 97.3, respectively. The R2 values for qua-
dratic models were also 99.2, 95.7, 97.5, 91.4 and 99.2 for
E. coli O157:H7, C. lipolytica, H. anomala, L. delbrueckii ssp.
bulgaricus and S. cerevisiae, consequently (Table 5).
Control samples at 10C were rated as 8.02 ± 1.25,
7.65 ± 0.96, 8.00 ± 1.05, 1.85 ± 0.40, 6.46 ± 0.46, 7.00 ± 0.58,
6.70 ± 0.33, 4.40 ± 0.72, 6.60 ± 0.63 and 6.70 ± 0.70 for
cloudiness/clearness, dullness/brightness, red color density,
particle status, density, odor/flavor, wine taste, bitter taste,
sour taste and after taste, and there was no significant differ-
ence among the control and PEF-processed samples for the
measured sensory attributes (P > 0.05) (Fig. 2).
It is an important factor that any processing method
applied to wine should not adversely affect its quality. The
quality of red wine depends on several factors including
TPSC, TAC, TMAC, color, aroma, sensory perception, etc.

Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc. 7
PEF PROCESSING OF RED WINE E.E. ABCA and G. AKDEMIR EVRENDILEK

TABLE 4. ANALYSIS OF VARIANCE RESULTS

Microorganism df Sum of squares Mean square F value P
FOR MICROBIAL INACTIVATION
Escherichia coli O157:H7
Efs 3 92.873 30.9578 327.44 0.000
Tt 2 9.962 4.9808 52.68 0.000
Efs × Tt 6 2.578 0.4296 4.54 0.001
Candida lypolitica
Efs 3 206.222 68.7408 1,338.49 0.000
Tt 2 56.934 28.4620 554.20 0.000
Efs × Tt 6 8.499 1.4166 27.58 0.000
Saccharomyces cerevisiae
Efs 3 294.089 98.0296 1,872.59 0.000
Tt 2 11.082 5.5409 105.84 0.000
Efs × Tt 6 11.153 1.8588 35.51 0.000
H. anomala
Efs 3 281.944 93.9815 1,378.32 0.000
Tt 2 5.837 2.9187 42.81 0.000
Efs × Tt 6 2.786 0.4644 6.81 0.000
Lactobacillus bulgaricus
Efs 3 156.998 52.3325 526.52 0.000
Tt 2 17.229 8.6497 87.03 0.000
Efs × Tt 6 8.496 1.4160 14.25 0.000

Efs, electric field strength; Tt, treatment temperature.

(Boulton 2001; Budić-Leto et al. 2006; Puertolas et al.
2010b). It was reported that phenolic compounds in red
wine exhibit antioxidant and free radical scavenging proper-
ties (Rice-Evans et al. 1996) that may play an important role
in human health, reducing the risk of various degenerative
diseases, such as cardiovascular diseases, osteoporosis and
cancer (Nichenametla et al. 2006; Puertolas et al. 2010a).
They make an essential contribution to the red wine
organoleptic attributes, particularly to color, bitterness,
astringency and mouthfeel (Boulton 2001). Wine color is
the first characteristic perceived by consumers; therefore, it
is one of the most important quality characteristics of red
wines. These compounds, in their monomeric form, are the
pigments responsible for the red color in red wines, and
they contribute to the stabilization of color during wine
aging, due to the development of red polymeric pigments
with other polyphenols (Boulton 2001; Budić-Leto et al.
2006).
The applicability of PEF to process red wine without
affecting important quality parameters is very important.
This study revealed that PEF processing with maximum of
31 kV/cm at 40 ± 2C can be successfully used to process red
wine samples with no significant change on pH, °Brix, TA,
color (L, a, b, hue and chroma), TMAC, TPSC and TAC as
well as sensory properties.
Previous studies related to PEF processing of wine
samples mostly focused on extraction of phenolics from
grape tissue, amelioration of red color for extraction, pro-
cessing of must by PEF and acceleration of aging process
(Garde-Cerdán et al. 2008; Lopez et al. 2009; Puertolas et al.

TABLE 5. ANALYSIS OF VARIANCE AND QUADRATIC MODEL RESULTS FOR MICROBIAL INACTIVATION

R2 for
F value P value R2 for model quadratic
Microorganisms for SRM for SRM validation Quadratic model model

Escherichia coli O157:H7 84.29 0.001 91.2 4.12 + 0.0937 temp − 0.00219 temp2 − 0.00302efs2 + 0.0473 99.2
efs − 0.00233 efs × temp
Candida lipolytica 23.78 0.001 91.6 7.37 − 0.209 temp + 0.00458 temp2 + 0.00150 efs2 − 0.166 95.7
efs − 0.00229 efs × temp
H. anomala 14.45 0.001 91.8 4.99 + 0.049 temp − 0.00149 temp2 + 0.00330 efs2 − 0.184 97.5
efs − 0.00298 efs × temp
Lactobacillus bulgaricus 8.58 0.001 92.0 6.57 − 0.026 temp + 0.00032 temp2 − 0.00131 efs2 − 0.0412 91.4
efs − 0.00196 efs × temp
Saccharomyces cerevisiae 33.06 0.001 97.3 5.76 + 0.0277 temp − 0.00073 temp2 − 0.00112 efs2 − 0.0846 99.2
efs − 0.00246 efs × temp

efs, electric field strength; SRM, surface response methodology; temp, temperature.

8 Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
E.E. ABCA and G. AKDEMIR EVRENDILEK
cloudiness/clearness
after taste
sour taste
FIG. 2. SENSORY ANALYSES OF RED WINE bitterness
SAMPLES PROCESSED BY PULSED ELECTRIC
FIELD AS A FUNCTION OF ELECTRIC FIELD
STRENGTH AND TREATMENT TEMPERATURE
typical wine taste
2010b). In most of the studies, the magnitude of applied
electric field is much lower than that of the study; therefore,
it is not really possible to compare the results obtained in
this study with the others. However, these studies also
revealed that PEF provided better extraction of phenolics,
better retention of color and more aroma compounds.
Concentration of metal ions is another important factor
for red wine quality. Increase in some metal ions such as Fe
and Cu can cause turbidity in wine. Moreover, ions such as
Fe, Cu and Al may cause metallic taste (Karadjova et al.
2002). The amount and type of ions can be changed by dif-
ferent factors, but migration from PEF treatment chamber
can also change the amount and type of metal ions. For all
the metal ions measured in the study, no significant differ-
ence was detected between the control and PEF-treated
samples. That means no significant migration occurred
from treatment chambers to red wine samples during PEF
processing.
Limited number of studies focused on inactivation of
microorganisms in wine. According to these studies PEF
processing at 35 kV/cm provided 4 log inactivation of
Kloeckera apiculata, S. cerevisiae, L. plantarum, L. hilgardii
and Gluconobacter oxydans (Marsellés-Fontanet et al.
2009). Inactivation of Dekkera anomala, D. bruxellensis,
L. plantarum and L. hilgardii in both must and wine
with 16–31 kV/cm electric field strength provided 3 log
reductions (Puertolas et al. 2009). Results showed that
L. delbrueckii ssp. bulgaricus was more resistant than E. coli
O157:H7 and bacteria were more resistant than yeasts.
The applicability of PEF processing on red wine shown in
this study revealed that red wine can be processed by PEF
without significantly affecting the important physical
parameters and sensory properties with significant inactiva-
tion of microorganisms. However, future studies need to be
Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
PEF PROCESSING OF RED WINE
0 kV/cm 10 C
0 kV/cm 30C
9
17 kV/cm 30C
8 dullness/brightness
7 24 kV/cm 30C
6 31 kV/cm 30C
5
4 red color intensity
3
density
odour/flavor
designed to search the effect of PEF processing on wine
aging, aroma compounds and sensory analyses to better
understand the effects of PEF.
ACKNOWLEDGMENTS
The authors would like to thank the Ministry of Develop-
ment State Planning Organisation (DPT, project no: DPT
2009 K 120 410) and Abant Izzet Baysal University Research
Fund (AİBU BAP project no: 2009.09.01.307) for financial
support, and Dimes Gida San ve Tic A.S. (Tokat, Turkey) for
providing wine samples.
REFERENCES
ANCÍN, C., GARDE, T., TORREA, D. and JIMENEZ, N. 2004.
Extraction of volatile compounds in model wine from
different oak woods: Effect of SO2. Food Res. Int. 4, 375–383.
ARAPITSAS, P., ANTONOPOULOS, A., STEFANOU, E. and
DOURTOGLOU, V.G. 2004. Artificial aging of wines using
oak chips. Food Chem. 86, 563–570.
BOULTON, R.B. 2001. The copigmentation of anthocyanins and
its role in the colour of red wine: A critical review. Am. J.
Enol. Viticul. 55, 67–87.
BUDIĆ-LETO, I., LOVRIĆ, T., GAJDOŠ KLJUSURIĆ, J., PEZO,
I. and VRHOVŠEK, U. 2006. Anthocyanin composition of the
red wine Babić affected by maceration treatment. Euro. Food
Res. Technol. 3–4, 397–402.
CORTÉS, C., ESTEVE, M.J., FRI´GOLA, A. and TORREGROSA,
F. 2005. Quality characteristics of horchata (a Spanish
vegetable beverage) treated with pulsed electric fields during
shelf-life. Food Chem. 91, 319–325.
COUTO, J.A., NEVES, F., CAMPOS, F. and HOGG, T. 2005.
Thermal inactivation of the wine spoilage yeasts
Dekkera/Brettanomyces. Int. J. Food Microbiol. 104, 337–344.
9
PEF PROCESSING OF RED WINE
CUBADDA, F., BALDINI, M., CARCEA, M., PASQUI, L.A.,
RAGGI, A. and STACCHINI, P. 2001. Influence of laboratory
homogenization procedures on trace element content of food
samples: An ICP-MS study on soft and durum wheat.
Food Addit. Contam. 18, 778–787.
DU TOIT, M. and PRETORIUS, I.S. 2000. Microbial spoilage
and preservation of wine: Using weapons from nature’s own
arsenal – a review. South African J. Enol. Viticult. 21, 74–92.
EVRENDILEK, G.A., ZHANG, Q.H. and RICHTER, R.E. 1999.
Inactivation of E. coli O157:H7 and E. coli 8739 in apple juice
by pulsed electric fields. J. Food Prot. 62, 793–796.
GARDE-CERDÁN, T., ARIAS-GIL, M.,
MARSELLES-FONTANET, A.R., ANCÍN-AZPILICUETA, C.
and MARTIN-BELLOSO, O. 2007. Effects of thermal and
non-thermal processing treatments on fatty acids and free
amino acids of grape juice. Food Cont. 18, 473–479.
GARDE-CERDÁN, T., MARSELLES-FONTANET, A.R.,
ARIAS-GIL, M., ANCÍN-AZPILICUETA, C. and
MARTIN-BELLOSO, O. 2008. Effect of storage conditions on
the volatile composition of wines obtained from must
stabilized by PEF during ageing without SO2. Innov. Food Sci.
Emerg. Technol. 9, 469–476.
GARDE-CERDÁN, T.G., GOÑI, D.T. and AZPILICUETA, C.A.
2004. Accumulation of volatile compounds during ageing of
two red wines with different composition. J. Food Eng. 65,
349–356.
JIN, Z.T. and ZHANG, Q.H. 2007. Pulsed electric field
inactivation of microorganisms and preservation of quality of
cranberry juice. J. Food Proc. Preserv. 23, 481–497.
KARADJOVA, I., IZGI, B. and GUCER, S. 2002. Fractionation
and speciation of Cu, Zn and Fe in wine samples by atomic
absorption spectrometry. Spectrochim. Acta 57, 581–590.
KNORR, D. and ANGERSBACH, A. 1998. Impact of
high-intensity electric field pulses on plant membrane
permeabilization. Trends Food Sci. Technol. 9, 185–191.
LEE, J., DURST, R.W. and WROLSTAD, R.E. 2005.
Determination of total monomeric anthocyanin pigment
content of fruit juices, beverages, natural colorants, and wines
by the pH differential method: Collaborative study. J. AOAC
Int. 88, 1269–1278.
LOPEZ, N., PUERTOLAS, E., CONDON, S., ALVAREZ, I. and
RASO, J. 2008. Effects of pulsed electric fields on the
extraction of phenolic compounds during the fermentation of
must of Tempranillo grapes. Innov. Food Sci. Emerg. Technol.
9, 477–482.
LOPEZ, N., PUERTOLAS, E., HERNANDEZ-ORTE, P.,
ALVAREZ, I. and RASO, J. 2009. Effect of a pulsed electric
field treatment on the anthocyanins composition and other
quality parameters of Cabernet Sauvignon freshly fermented
model wines obtained after different maceration times.
LWT – Food Sci. Technol. 42, 1225–1231.
MARSELLÉS-FONTANET, A.R., PUIG, A., OLMOS, P.,
MINGUEZ-SANZ, S. and MARTÍN-BELLOSO, O. 2009.
Optimising the inactivation of grape juice spoilage organisms
by pulsed electric fields. Int. J. Food Microbiol. 130, 159–165.
10 Journal of Food Processing and Preservation •• (2014) ••–•• © 2014 Wiley Periodicals, Inc.
E.E. ABCA and G. AKDEMIR EVRENDILEK
MARTIN-BELLOSO, O., VEGA-MERCADO, H., QIN, B.L.,
CHANG, F.J., BARBOSA-CÁNOVAS, G.V. and SWANSON,
B.G. 1997. Inactivation of Escherichia coli suspended in liquid
egg using pulsed electric fields. J. Food Proc. Preserv. 21,
193–208.
MOON, J.-A. and TERAO, J. 1998. Antioxidant activity of
caffeic acid and dihydrocaffeic acid in lard and human
low-density lipoprotein. J. Agric. Food Chem. 46,
5062–5065.
MORALES, M.L., BENITEZ, B. and TRONCOSO, A.M. 2004.
Accelerated aging of wine vinegars with oak chips: Evaluation
of wood flavour compounds. Food Chem. 88, 305–315.
NAKAI, S., LI-CHEN, E.C.Y. and DOU, J. 2006. Experimental
design and response surface methodology. In Handbook of
Food and Bioprocess Modelling Techniques, Vol. 1 (S. Sablani,
A. Data, M.S. Rahman and A. Mujumdar, eds.) pp. 293–323,
CRC Press, Boca Raton, FL.
NICHENAMETLA, S., TARUSCIO, T., BARNEY, D. and EXON,
J. 2006. A review of the effects and mechanisms of
polyphenolics in cancer. Crit. Rev. Food Sci. Nutr. 46,
161–183.
PUERTOLAS, E., LOPEZ, N., CONDON, S., RASO, J. and
ALVAREZ, I. 2009. Pulsed electric fields inactivation of wine
spoilage yeast and bacteria. Int. J. Food Microbiol. 130, 49–55.
PUERTOLAS, E., SALDANA, G., ALVAREZ, I. and RASO, J.
2010a. Effect of pulsed electric fields processing on red wine
chromatic and phenolic characteristics during aging in oak
barrels. J. Agric. Food Chem. 58, 2351–2357.
PUERTOLAS, E., SALDANA, G., CONDON, S., ALVAREZ, I.
and RASO, J. 2010b. Evolution of polyphenolic compounds in
red wine from Cabernet Sauvignon grapes processed by
pulsed electric fields during aging in bottle. Food Chem. 119,
1063–1070.
RICE-EVANS, C.A., MILLER, N.J. and PAGANGA, G. 1996.
Structure-antioxidant activity relationships of flavonoids and
phenolic acids. Free Rad. Biol. Med. 20, 933–956.
RIVAS, A., RODRIGO, D., MARTÍNEZ, A.,
BARBOSA-CÁNOVAS, G.V. and RODRIGO, M. 2006. Effect
of PEF and heat pasteurization on the physical-chemical
characteristics of blended orange and carrot juice.
LWT – Food Sci. Technol. 39, 163–1170.
SALE, A.J.H. and HAMILTON, W.A. 1967. Effects of high
electric fields on microorganisms I. Killing of bacteria and
yeast. Biochim. Biophys. Acta 148, 781–788.
SPANOS, G.A. and WROLSTAD, R.E. 1990. Influence of variety,
maturity, processing and storage on the phenol composition
of pear juice. J. Agric. Food Chem. 38, 817–824.
TIWARI, B.K., O’DONNELL, C.P. and CULLEN, P.J. 2009. Effect
of non thermal processing technologies on the anthocyanin
content of fruit juices. Trends Food Sci. Technol. 20, 137–145.
USSEGLIO-THOMASSET, L. 1992. Properties and use of
sulphur dioxide. Food Addit. Contam. 9, 399–404.
ZIMMERMAN, U., PILWART, G. and RIEMANN, F. 1974.
Dielectric breakdown in cell membranes. Biophys. J. 14,
881–889.