Pulsed Electric Fields in Wineries: Potential

Applications
Guillermo Saldaa, Elisa Luengo, Eduardo Purtolas, Ignacio
lvarez, and Javier Raso

Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Application of PEF for Improving Must Expression by Pressing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Application of PEF for Improving Red Winemaking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Studies Conducted Applying PEF in Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Studies Conducted Applying PEF in Continuous Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Application of PEF for Improving Waste Recovery from Wineries . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Application of PEF for Inactivation of Wine Spoilage Microorganisms . . . . . . . . . . . . . . . . . . . . . . . .
Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Pulsed electric eld is an innovative processing technology that may improve

different operations conducted in wineries. Wineries may take advantage of the
ability of PEF to inactivate microorganisms or to electroporate the cell membrane
of plant tissues improving extraction of compounds of interest from grapes but
also from residues generated by the wineries. Electroporation of white grapes by
PEF may improve the yield of must expression from white grapes. The
G. Saldaa (*) I. lvarez J. Raso
Department of Animal Production and Food Science, Tecnologa de los Alimentos, Facultad de
Veterinaria, Universidad de Zaragoza, Zaragoza, Spain
e-mail: gsaldana@unizar.es; ialvalan@unizar.es; jraso@unizar.es
E. Luengo
School of Agriculture and Food Science, University College Dublin, Dublin, Ireland
e-mail: elisa.luengo-maranillo@ucd.ie
E. Purtolas
Food Research Division, AZTI, Derio, Bizkaia, Spain
e-mail: epuertolas@azti.es
# Springer International Publishing AG 2016
D. Miklavcic, Handbook of Electroporation,
DOI 10.1007/978-3-319-26779-1_155-1

G. Saldaa et al.

permeabilization by PEF of red grape skin cells permits reducing the duration of
the maceration through vinication or to increase the color and concentration of
anthocyanins and polyphenolic compounds in the wine without impairing its
sensorial attributes. On the other hand, PEF pretreatments have potential for
improving solvent extraction of polyphenols from winery waste. Finally, the
capability of PEF to inactivate spoilage microorganisms preserving physicochemical and sensorial properties of must and wines may contribute to enhance
the quality of wine by ensuring reproducible fermentations or by reducing or
replacing the SO2 in winemaking. This chapter provides an overview of the
research conducted so far on applications of PEF in wineries remarking the
most important challenges of the future application of PEF in the winemaking
process.
Keywords

Pulsed electric elds Wine Extraction Microbial inactivation Juice expression Polyphenolic compounds

Introduction
Grapes represent one of the most important fruit crop worldwide with an annual
world production of around 70 million tons. Although grapes may be processed into
a variety of different food products such as grape juice, jams, or raisins, approximately 80 % of the grape production worldwide is dedicated to winemaking.
Wine is a beverage obtained by alcoholic fermentation of the sugars of the juice of
grapes by yeast. Fermentation can be conducted by wild yeasts that are normally
present on the grapes or fermentation tanks can be inoculated with commercial
preparations of yeast to prevent unpredictable fermentations. Winemaking of white
and red wine diverges from the rst processing steps when the grapes are taken into a
winery (Fig. 1). White wine is made by fermenting juice which is obtained by
pressing white grapes. However, red wine is obtained from the must of red or
black grapes (mixture of crushed grapes, skins, juice, and seeds) that undergoes
fermentation together with the grape skin. In this step, called macerationfermentation, yeasts convert the sugars of the must into ethanol but also polyphenolic compounds are extracted from the grape skin. Polyphenols contribute substantially to the quality of red wines because they affect their color, avor stability, and
aging behavior. Furthermore, phenolic compounds are also associated with the
benecial physiological effects deriving from moderate wine consumption. Red
wine is sometimes stored in oak barrels for aging for a period of time of weeks to
months to provide some aromas and to obtain a smoother wine. In the case of ros
wines, fermentation is conducted from juice of red grapes obtained by long enough
maceration with the grapes skins to obtain the characteristic color of this type of
wines.

Pulsed Electric Fields in Wineries: Potential Applications

Fig. 1 Red and white winemaking steps

Technological innovation represents a driving force to maintain and enhance the

competitiveness of food industry. The development of innovations able to impact the
market are increasingly attracting for wineries once they can improve wine quality
and processing sustainability. Pulsed electric eld (PEF) technology is an innovative
processing technology with potential applications in wineries for different purposes.
During PEF processing, a pumpable product is passed through a treatment chamber

G. Saldaa et al.

where it is subjected to short pulses (s) of high voltage (kV). The applied external
voltage generates an electric eld whose strength depends not only on the voltage
intensity but also on the distance between the electrodes. When exposed to a
sufciently strong electric eld, the cell membrane undergoes an electrical breakdown, which renders it permeable to molecules that are otherwise unable to cross
it. This phenomenon is called membrane electroporation. If the intensity of the
electric eld is not high enough or if the exposure to the electric eld is sufciently
short, the membrane can spontaneously return to its initial state, remaining viable
(reversible electroporation). However, intense electric elds or longer expositions
can cause irreversible electroporation.
Applications of PEF in the food industry for improving mass transfer or for
microbial inactivation are based on the irreversible electroporation of the cell
membranes. The mass transfer phenomenon through cell membranes occurs in
many operations of the food industry that aims at obtaining a given intracellular
compound of interest, removing water from foods (drying) or introducing a given
substance into the food matrix. The breakdown of the cell membranes by different
techniques such as grinding, heating, or enzymatic maceration is a common
pretreatment step to improve mass transfer rates. Irreversible electroporation of the
cytoplasmatic membrane is an effective alternative to these pretreatments especially
when the complete disintegration of cell membranes is not desired. On the other
hand, the microbial cytoplasmic membrane acts as a semipermeable barrier, playing
an essential role in maintaining microbial homeostasis. The electroporation of the
cytoplasmic membranes of bacteria, yeast, and molds causes the loss of their
selective permeability, leading to microbial death (Garca et al. 2006). The capability
of PEF to inactivate vegetative cells of pathogenic and spoiling microorganisms at
temperatures that avoid the harmful effect of heat on the organoleptic properties and
nutrient value of foods is very attractive for the food industry which demands gentle
methods for food decontamination.
Application of PEF has been investigated over the last decades in different elds
including improving different operations conducted in wineries. Wineries may take
advantage of the ability of PEF to electroporate the cell membrane of plant tissues
improving extraction of compounds of interest from grapes but also from residues
generated by the wineries and to inactivate microorganisms at temperatures not
affecting the avor and aroma characteristic of the wine. This chapter provides an
overview of the research conducted on the main potential application of PEF in
wineries.

Application of PEF for Improving Must Expression by Pressing

Extraction by pressing called also solidliquid expression is an operation widely
used in the wineries in the elaboration of different types of wine. In white and ros
winemaking, grapes are pressed in order to separate juice. On the other hand, after
the fermentation of the red wine, the grape pomace that have been in contact with the
juice in the fermentation tanks are pressed to extract the remaining wine. Different

Pulsed Electric Fields in Wineries: Potential Applications

studies have investigated the effect of PEF on juice extraction of white grapes;
however, the effect of PEF in juice expression of red grapes or in wine extraction
from grape pomace after fermentation has not been assayed.
Juice extraction from white grapes is a critical step that has a large inuence on
the nal quality of the white wine. In order to obtain a high-quality juice for white
winemaking, pressing at moderate pressure is usually led in order to obtain low
turbidity which depends on the content of solid particles in suspension and low
concentration of color pigments. Furthermore, short pressing times are conducted to
avoid juice browning by polyphenol oxidation.
The effect on the extraction yield and quality of white grapes juice after the
application of PEF treatments before pressing of the grapes have been investigated,
although white wine from juice obtained from PEF-treated grapes has not been
produced.
A laboratory lter-press equipped with two electrodes that permitted applying
moderate electric eld strengths (0.251 kV/cm) before pressing or after a given
pressing time was used to investigate the effect of electroporation on yield and
characteristics of juice extracted from different white grapes (Muscadelle,
Sauvignon, and Semillon) (Praporscic et al. 2005). Working at a pressure of
5 bars, it was observed that the application of the PEF treatment to the white grapes
before pressing was more effective than the application of the PEF treatment to the
grapes after a pressing period. A PEF treatment with pulses of duration in the range
of milliseconds and a total duration of 0.3 s at a low electric eld (0.75 kV/cm)
increased the juice yield extraction by 24 % as compared with the untreated grapes
(Praporscic et al. 2005). The electroporation of the cells of the three varieties
investigated allowed to obtain a juice that had a lower turbidity and absorbance at
520 nm. In another study conducted with the same laboratory lter-press, the effect
of the previous electroporation of Chardonnay grapes on the juice extraction yield
using two extraction regimes was investigated: constant pressure (0.51 bar) and
progressive pressure (increasing the pressure up to 1 bar) (Grimi et al. 2009). In this
case, no signicant effect on turbidity and polyphenol content was observed when
the PEF treatment was applied to Chardonnay grapes before pressing at a constant
pressure of 1 bar but juice yield increased by 18 %. When a progressive pressureincrease regime was applied, the most remarkable PEF effect was the rise of
polyphenol content of the must by 15 %. This increment on the content of polyphenols could be of interest for obtaining white grape juice but probably not for the
elaboration of white wine.
The light pressing conditions required for obtaining a high-quality white grape
juice may be insufcient for the effective rupture of the membranes of the cells
where the juice is enclosed. The electroporation by PEF of the cells of the white
grapes before pressing may permit obtaining a high amount of juice with a quality
similar or improved with respect to the untreated grapes or perhaps to reduce the
intensity and/or duration of pressurization maintaining extraction yields and improving the quality of the juice. The promising results obtained at laboratory scale
concerning to the expression of white juice should be conrmed at higher scale,

G. Saldaa et al.

and white wine should be elaborated with the grape juice obtained from PEF-treated
grapes to evaluate the inuence of the treatment on wine quality.

Application of PEF for Improving Red Winemaking

Red wine quality is strongly affected by the phenolic compounds which are responsible for the sensory characteristics such as color and taste and aging properties. On
the other hand, in vitro and in vivo studies have showed that some phenolic
compounds of red wine have antioxidant properties which may play a positive role
in human health including protection against cardiovascular diseases and cancer
(Nichenametla et al. 2006).
Wine phenolic compounds can be divided in nonavonoid (phenolic acids and
stilbenes) and avonoid (anthocyanins, avan-3-ols and avonols). The monomeric
forms of anthocyanins are mainly responsible for the red color of young red wines,
and they also contribute to the development of red polymeric pigments during wine
aging by association between anthocyanins pigments and other polyphenols especially tannins and phenolic acids. Flavan-3-ols (tannins) are a large family of
polyphenolic compounds which are mainly responsible for the astringency, bitterness, and structure of wines. The last group of avonoids is avonols that contribute
to bitterness, display antioxidant activity, and affect red wine color acting as
copigments of anthocyanins. Between nonavonoid compounds, stilbenes have
been also recognized as compounds with antioxidant and anti-inammatory
properties.
Anthocyanins and tannins are responsible for the major sensorial characteristics
of red wine. Red grape skins are particularly rich in these phenolic compounds but
also a part of the tannins of red wine proceeds from the seeds. Grape skin consists of
an external cuticle, the epidermis, and the hypodermis. The hypodermis is the layer
closest to the pulp and is composed of several layers that contain most of the grape
skin phenols. While anthocyanins are located in the upper cell layers of the hypodermis, tannins are located into the vacuoles of hypodermal cells.
The phenolic composition of wines is conditioned by the grape variety and by
other factors that affect the berry development such as soil, geographical location,
and weather conditions. On the other hand, the winemaking practices also play an
important role in the extraction of polyphenols from the grape skins. It has been
observed that during winemaking only about 40 % of anthocyanins and 20 % of
tannins of the grape skins are transferred to the wine. This limited extraction is a
consequence of the low permeability of the cell walls and cytoplasmic membranes of
the hypodermal cells to these compounds.
Different strategies have been developed in wineries to enhance the extraction of
phenolic compounds during the fermentation-maceration step of red winemaking
because of the large inuence of these compounds in sensory and nutritional quality
of red wine (Bautista-Ortn et al. 2007; Sacchi et al. 2005). The most traditional
procedure for obtaining wines with high phenolic content is extending the maceration time for 34 weeks representing a period of time much longer than those

Pulsed Electric Fields in Wineries: Potential Applications

Fig. 2 Effect of PEF on the extraction of grape polyphenols after 1 h of maceration

required for fermentation (12 weeks) or increasing the fermentation temperature.

However, some problems have been identied when these enological practices are
used. Longer maceration time reduces the production capacity of the wineries or
requires increasing the number of fermentation tanks to maintain productivity.
Conducting fermentation at higher temperature may cause stopping fermentation
and loss of volatile compounds. On the other hand, extraction of polyphenols is
improved by using different techniques that increases the permeability of the envelopes of the grape skin cells in order to facilitate their release such as thermovinication, grape freezing, ash-release, or the use of pectolytic enzymes. However,
some of these techniques require high energy consumption and may affect the
quality of red wines. The use of commercially pectolytic enzymes that act on the
wall of the grape skin cells is a widespread practice in the wineries to improve the
color and aroma of wines. However, some contradictory results attributed to the
different nature and activities of the commercial enzymes preparations, and the
presence of unwanted enzymes in the preparations such as -glucosidase have
been reported (Bautista-Ortn et al. 2005).
The application of PEF to accelerate and/or increase the extraction of phenolic
compounds during the maceration-fermentation step of red winemaking has been
deeply investigated. In some of these studies, it has been observed that the electroporation of the grape skin cells also improves the extraction of aromatic compounds
(Delsart et al. 2012; Garde-Cerdn et al. 2013). This electroporation can be achieved
using moderate electric elds (0,51 kV/cm) and treatment times in the range of
40100 ms or higher electric elds (110 kV/cm) and shorter treatment times
(100 s-1 ms).

G. Saldaa et al.

Studies Conducted Applying PEF in Batch

The potential of PEF for improving extraction of polyphenols during red
winemaking was reported by rst time by Lpez et al. (2008) (Fig. 2). These authors
used a batch parallel electrode treatment chamber to apply to the PEF treatments and
the effect of the treatment was evaluated along the maceration-fermentation time in
0.5 L ask containing 150 g of Tempranillo grape skins. A signifcant improvement
in the release of phenolic compounds was observed when grapes were previously
treated by PEF (Fig. 2). The color intensity (CI), the anthocyanin content (AC), and
the total polyphenol index (TPI) at the end of the fermentation of red wine that
contained skins treated at 10 kV/cm, 6.7 kJ/kg were a 23 %, 26 %, and 24 % higher,
respectively, than in the control sample. On the other hand, no signicant effect of
the PEF treatment was observed in the other physicochemical properties of red wine.
These rst results indicated that electroporation of the grape skins preceding the
maceration step could contribute to reduce the duration of the maceration during
vinication or to increase the CI, AC, and TPI in the nal wine.
Similar effects of PEF treatment applied in batch treatment chambers of parallel
electrode with a capacity from few grams to 4.5 kg has been reported for different
varieties of grapes such as Cabernet Sauvignon, Merlot, Syrah, Grenache, Mazuelo,
Graciano, Aglianico, and Piedirosso harvested in different countries such as Spain,
France, Lebanon, and Italy. The improvements obtained by application of PEF
treatments are dependent on the intensity of the treatment and on the grape variety.
Generally the PEF treatments were more effective at higher electric eld strengths,
although in some varieties such as Graciano the most suitable treatment was the
lowest intensity applied (in the range of 210 kV/cm) and in Cabernet Sauvignon it
was an intermediate intensity (5 kV/cm). These results indicate that for some grape
varieties, an intense electroporation of the skin cells may promote a very intense
release of phenolic compounds that precipitate rather than keep stabilized in the
wine. The different effect of PEF on the extraction of polyphenols for different grape
varieties has been explained in terms of polyphenol extractability that depends on
grape maturity, cell morphology, and composition of the skin cell wall. Results
obtained suggest that the electroporation of the cells of the grape skin was more
useful in those situations in which the extraction of phenolic compounds from the
grape skins are more difcult.
The potential of PEF for improving red winemaking has been also compared to
other innovative treatments such as ultrasound and common technologies used in
wineries to improve phenolic extraction such as the addition of enzymes and
thermovinication treatments (El Darra et al. 2013a, b, 2016). Results obtained
showed that the pretreatment of the grapes of the variety Cabernet Franc by
ultrasound, thermovinication, and PEF improved polyphenol extraction (anthocyanins and tannins content), color intensity, and scavenging activity of the samples
during fermentation. However, the PEF treatments of similar total specic energy
(around 50 kJ/kg) applied at moderate (0.8 kV/cm, 100 ms) and higher (5 kV/cm,
1 ms) intensity were found to be the most effective pretreatments in terms of
phenolic extraction and energy consumption (El Darra et al. 2013a, b). Similar

Pulsed Electric Fields in Wineries: Potential Applications

conclusions were obtained when it was compared to the effects of different pretreatments of the Cabernet Sauvignon variety grapes with PEF, enzymes, and
thermovinication. The PEF treatment allowed to obtain a new prole of red wine
in terms of color attributes and polyphenol content without addition of additives such
as enzymes and requiring less energy consumption (48 kJ/kg) compared to
thermovinication (418 kJ/kg) (El Darra et al. 2016).
In studies in which the anthocyanins were analyzed by high performance liquid
chromatography (HPLC), it was observed that the PEF treatment did not affect the
HPLC prole of the wine after alcoholic fermentation (Lpez et al. 2009) and after
6 months of bottling (Delsart, et al. 2013) indicating that the enhancement in the
extraction of anthocyanins by PEF was not selective for any specic anthocyanin
molecule. Similar results were observed when it was compared after 6 month of
bottling the concentration of monomers and dimers of avan-3-ols analyzed by
HPLC in wines obtained from untreated and PEF-treated grapes of Cabernet
Sauvignon (Delsart et al. 2013).
The effect of treating the grapes by PEF before red wine vincation on the
organoleptic characteristics of the wine by sensory evaluation has been also evaluated (Delsart et al. 2012). A panel of professional testers preferred Merlot wine
obtained by grapes treated by PEF at 0.5 and 0.7 kV/cm for 40 ms than control wine
or wine obtained by grapes after an intense PEF treatment (0.7 kV/cm, 100 ms).
While after applying the most intense treatment, the diffusion of tannins to the wine
was probably excessive; in the case of less-intense PEF treatments, the diffusion of
these compounds was lower and the wines were more aromatic and fruity suggesting
that PEF treatments promote an additional diffusion of aromatic compounds from
the skin cells.

Studies Conducted Applying PEF in Continuous Flow

The implementation of the PEF technology for improving red winemaking requires
the application of the treatments in continuous ow. Continuous ow process
permits to process big amount of grapes to perform the maceration-fermentation
step and obtaining the required quantity of wine to be able to evaluate if the benets
observed in the studies conducted in batch at laboratory scale are maintained during
wine aging.
The feasibility of processing red grape by PEF in continuous ow (118 kg/h)
using a colinear treatment chamber and the inuence of the PEF treatment on the
evolution of chromatic parameters and phenolic content during winemaking and
aging in bottle and in oak barrels were investigated by Purtolas et al. (2010b) using
Cabernet-Sauvignon grapes. Wine was elaborated in tanks of 80 l, and in order to
evaluate the potential of the PEF technology for reducing maceration time, the grape
skins were separated from the fermenting must after 96 h while in the control the
grape skins were in contact with the fermenting must until the end of fermentation
(144 h). Although the maceration time for wine obtained from Cabernet-Sauvignon
grapes treated by PEF (5 kV/cm, 150 s, 3.67 kJ/kg) was 48 h shorter, the wine at the

10

G. Saldaa et al.

end of the alcoholic fermentation presented higher CI, AC, and TPI than the control
wine. Differences between wine obtained from PEF-treated and PEF-untreated
grapes were also observed after aging the wine in bottle. After 4 months, the CI,
AC, and TPI, of the wine obtained from PEF-treated grapes, were 27 %, 18 %, and
10 % higher, respectively, than in the control wine (Fig. 3). On the other hand, no
differences were observed in the HPLC polyphenolic proles of both wines, indicating that the PEF treatment did not produce a selective effect on the extraction of
any specic polyphenol. Results of a sensorial evaluation indicated that PEF wine
did not have any strange taste or off-avors that could be caused by the PEF
treatment. The better chromatic characteristics and higher polyphenolic content
obtained from the PEF treatment after the fermentation process remained or even
increased during aging in American oak barrels for 6 month and their posterior
storage in bottles for 8 months (Purtolas et al. 2010b).
The effect of PEF applied in continuous ow (400 kg/h) on improving the
phenolic compound extraction from three grape varieties (Graciano, Tempranillo,
Grenache) during two vintages has been also studied (Lpez-Giral et al. 2015). This
investigation not only conrms that the PEF effect depends on the grape variety but
also that grape physicochemical composition, that it was different for each vintages,
inuences PEF effect. The improvement in the extraction of polyphenols from the
grapes treated by PEF was more important when the concentration of phenolic
compounds in the skins was lower. Therefore, the application of the PEF technology
in the wineries for improving extraction of polyphenols would be particularly
interesting in those vintages in which the concentration of these compounds in the
grape skins is poor.
Studies conducted by the same research group demonstrated that in the three
varieties, the treatment of the grapes by PEF increased the stilbene content including
resveratrol and piceid in the must up to 200 % in Tempranillo, 60 % in Greneche, and
50 % in Graciano (Lpez-Alfaro et al. 2013). Resveratrol is a polyphenol belonging to
the stilbene family, the most abundant glycosylated form of this molecule (piceid) in
grapes. Resveratrol is one of the most widely investigated phenol in wines due to the
benecial properties attributed to this compound such as cardioprotective, anticancer,
antidiabetic, neuroprotective, and antiaging activities.
The enhancement of the aromatic characteristics of the wine by PEF previously
observed for the wine of Merlot variety in experiments conducted in batches was
conrmed for the wine of the Grenache variety obtained from grapes treated by PEF
in continuous ow by analyzing the volatile compounds by solid-phase
microextraction gas chromatography mass spectrometry(SPME-GC-MS). The PEF
treatment enhanced the aromatic composition of Grenache wine by increasing the
quantity of monoterpenoids, -ionone, total esters, and benzenoid compounds.
However, the volatile composition of Tempranillo and Graciano wines was not
improved by treating the grapes by PEF (Lpez-Alfaro et al. 2013).
Trials conducted in a small winery in which 6000 kg of grapes of Grenache
variety were PEF treated (4.3 kV/cm, 60 s) using a colinear treatment chamber at a
ow of 1900 kg/h conrmed results obtained at laboratory and pilot plant scale
(Luengo et al. 2014). Wine obtained from PEF-treated grapes with a maceration time

Pulsed Electric Fields in Wineries: Potential Applications

11

30

Colour intensity

25
20
15
10
5
0
EAF

EMF

4B

EAF

EMF

4B

EAF

EMF

4B

1400

Anthocyanin (mg/L)

1200
1000
800
600
400
200
0

Total polyphenol index

70
60
50
40
30
20
10
0

Fig. 3 Evolution of color intensity (a), anthocyanin content (b), and index of total polyphenols (c)
during vinication and maturation of untreated and PEF-treated Cabernet Sauvignon grapes. EAF: end
of alcoholic fermentation, EMF: end of malolactic fermentation, B: bottling, B4: after 4 months of aging
in bottle (Adapted from Puertolas et al. 2010a)

12

G. Saldaa et al.

of 7 days was compared with wine obtained from untreated and PEF-treated grapes
with the current maceration time used in the winery (14 days). After 7 days of
maceration, the CI, AC, and IPT of the wine obtained from grapes treated by PEF
were 12.5 %, 25 % and 23.5 % higher, respectively, than in wine obtained from
untreated grapes after 14 days of maceration. However, after 14 days of maceration
no signicant differences were observed between the control wine and the wine
obtained from grapes treated by PEF for these three indices. These results conrm
the potential of PEF to obtain wine with a sufcient concentration of polyphenolic
compounds with moderate maceration times. A sensory analysis revealed that the
wine obtained from PEF-treated grapes with a maceration of 7 days was more
signicantly preferred (95 % CL) than the control wine obtained with a longer
maceration time (14 days).
Results obtained in the studies conducted to evaluate the potential of PEF for
improving red winemaking indicate that the low energy consumption (0.46.7 kJ/kg)
and the short processing time (<1 s) required for the electroporation of grape skin
cells are key advantages of this technology for obtaining wines with a higher content
of polyphenolic compounds and with better chromatic and sensorial characteristics
and for reducing the time duration of maceration during vinication.

Application of PEF for Improving Waste Recovery from Wineries

Winemaking is currently one of the most important agricultural activities in the
world that generates a big amount of wastes such as grape pomace, the solid fraction
that remains after pressing. In white and ros winemaking, the seeds and skins of
grapes are pressed to obtain the must to be fermented, but in red winemaking, the
pomace consists of these solids that have been in contact with the fermenting must.
Grape pomace has been traditionally used as a soil conditioner or as a feed for
animals. However, in recent decades, grape pomace has been recognized as a
valuable source of good and cheap phenolic compounds. During red winemaking,
about 60 % of anthocyanins and 20 % of tannins of the grape skin remain in the skin.
In the last years, considerable effort has been conducted to valorize winery waste for
extracting polyphenols to be used as food colorants of natural origin or dietary
supplements due to their antioxidant properties.
The potential of PEF for enhancing extraction of polyphenols enclosed in skin or
seed cells by conventional solvent extraction that is the technique generally used at
industrial scale for extracting bioactive compounds from plant matrices has been
studied. The application of PEF treatment (3 kV/cm, 30 pulses, 10 kJ/kg) to the
grape skin waste of the Dornfelder variety enhanced the total phenolic content and
the anthocyanin concentration in a mixture of ethanol and water (50/50 v/v) heated at
70  C by 100 % and 17 %, respectively (Corrales et al. 2008). The antioxidant
activity of the extracts was fourfold higher for the samples treated by PEF. In the case
of extraction of polyphenols from the skin of a white grape variety (Chardonnay), the
application of a PEF treatment at lower electric eld strength (1.3 kV/cm) but with a

Pulsed Electric Fields in Wineries: Potential Applications

13

higher specic energy (120 kJ/kg) increased the concentration of polyphenols by

10 % after 60 min of extraction at 20  C in water (Boussetta et al. 2009).
Different pressures (010 bars) were applied to the skin of grapes of Dunkelfelder
variety after maceration-fermentation in order to assess the effect of the density
(0.61.3 g/cm3) on PEF efciency (Brianceau et al. 2015). After the application of
the treatment, the polyphenol extraction was conducted in a mixture of ethanol and
water (50/50 %) at different temperatures (2050  C). The highest efciency of the
PEF treatment (1.2 kV/cm, 18 kJ/kg) was obtained when the density of the grape
pomace was 1.0 g/cm3 that was attained by compression at 2 bars. Under these
optimal processing parameters, the PEF treatment increased the extraction of total
polyphenols regardless of the extraction temperatures. For example, at 20  C the
total quantity of additionally extracted polyphenols increased by 15 %. On the other
hand, this treatment increases the extraction of total anthocyanins in the range of
5.8 % (50  C) to 18.9 % (20  C). The effect of PEF (13.3 kV/cm, up to 564 kJ/kg) on
the recovery of anthocyanins from fermented pomace of the same grape variety was
compared with other alternative physical techniques such as ultrasound (US) and
high-voltage electrical discharges (HVED) (Barba et al. 2015). PEF remarkably
increased the extraction yield of total anthocyanins up to 22 % and 55 % in
comparison with US and HVED, respectively.
Grape seeds are the most studied and exploited residues from the wineries due to
the amount of these residues generated in each harvest (3852 % of grape) and its
high content of phenols, mainly avonols and tannins. The electroporation by PEF
of the cells of the grape seed of Pinot Meunier required higher electric eld strengths
(8 to 20 kV/cm) than those required for electroporation of the skin grape cells
(Boussetta et al. 2012). The improvement in the polyphenols extraction by the
application of a PEF treatment to the grape seeds was more remarkable when the
extraction temperature was increased until 50  C in presence of ethanol (3050 %).
A PEF treatment at 20 kV/cm for 20 ms increased the polyphenol extraction yield
more than three times in water and more than four times in ethanol (30 %) after
60 min of extraction at 50  C.
Studies conducted have demonstrated that PEF pretreatments have potential for
improving solvent extraction of polyphenols from winery waste. However, for future
industrial implementation the current studies that have been only focused on the
improvement of the extraction yield should be complemented with additional investigations to demonstrate other potential benets derived from the electroporation of
the cells such as reducing or avoiding the use of organic solvents and reducing
energy consumption by shortening the extraction time and/or decreasing the extraction temperature.

Application of PEF for Inactivation of Wine Spoilage

Microorganisms
Winemaking is inherently a microbial process wherein components of the grape are
transformed by yeast and bacteria into avor and aroma characteristics of the wine.
The primary fermentation that occurs in winemaking is bring about by the yeast

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G. Saldaa et al.

Saccharomyces (S. cerevisiae and S. bayanus are commonly found in wine fermentations) that metabolizes the sugars producing ethanol. Fermentation can be
conducted by autochthonous or native yeasts which are originally present in the
must or by commercial active dry preparation of a yeast strain with key enological
properties. Autochthonous or native fermentations can be at high risk for the
development of off-odors or incomplete fermentations. In order to assure correct
fermentation completion, sulfur dioxide is added to prevent the growth of
non-Saccharomyces yeast and bacteria that are more intolerant to this compound
than Saccharomyces.
In the elaboration of red wines and white wine with aging step, generally a second
fermentation occurs in which lactic acid bacteria convert malic acid into lactic acid
during malolactic fermentation. When both fermentation processes are nished,
microbial populations should be reduced to avoid post-fermentation processes that
may negatively affect the wine organoleptic characteristics.
Similarly to other food industries, the development of spoilage microorganisms is
one of the most important problems causing great economic losses in wineries. The
growth of undesirable yeast and bacteria may impair the development of the
inoculated yeast added to the must for alcoholic fermentation and may also produce
important sensorial changes in the wine. For example, acetic acid bacteria are able to
spoil the wine by ethanol acidication, lactic acid bacteria are responsible for the
alteration named piqre lactique, and yeast from the genus Brettanomyces are
involved in the formation of unpleasant odors in the wine which are described as
leather, animal, and horse sweat.
As thermal process may cause undesirable effect on wines, the addition of sulfur
dioxide (SO2) is the common practice used in wineries to decrease the risk of
microbial spoilage during the winemaking process. However, the microbial sensitivity to SO2 varies considerably between strains of microorganisms, and there is a
worldwide trend to reduce SO2 levels in wine due to its possible negative effects in
the health of consumers with a special sensitivity (Purtolas et al. 2009).
Several studies have demonstrated that PEF is an effective technology to inactivate bacteria and yeast in must and wine. The application of PEF as an alternative to
the addition of SO2 as antimicrobial compound was investigated using Parellada
white grape variety (Garde-Cerdn et al. 2008). These authors demonstrated that
when grape must was treated by PEF before fermentation, the SO2 concentration
could be reduced to safer levels or even eliminated. PEF treatments together with the
inoculation of a starter strain of yeast could lead to obtain a reproducible fermentation of must without modifying signicantly the composition of volatile compounds
responsible for the typical avor of wines through alcoholic fermentation and aging
of white wine.
The potential of PEF technology to control wine spoilage microbiota such as
Dekkera anomala, Dekkera bruxellensis, Lactobacillus plantarum, and Lactobacillus hilgardii has been investigated in must and wine (Purtolas et al. 2009). Yeasts
were more PEF sensitive than bacteria. At 29 kV/cm, a treatment of a specic energy
of around 150 kJ/kg and 186 kJ/kg was required to inactivate 3 log10 cycles the
population of the two Dekkera strains and the two Lactobacillus strains, respectively.

Pulsed Electric Fields in Wineries: Potential Applications

15

Considering the normal variation in concentration of these spoilage microorganisms

in must and wine, this inactivation could be enough to avoid the contamination of the
processing contact surfaces and controlling the development of alterations during the
wine aging in barrels and storage in bottles.
The most complete study on inactivation of wine-associated microbiota by PEF
was conducted by Gonzlez-Arenzana et al. (2015). Four different PEF treatments in
continuous process were tested in a continuous-ow system for inactivating 25 species of yeast, lactic acid bacteria, and acetic acid bacteria associated with
winemaking. The inactivation obtained was highly dependent on the microorganism
investigated. Overall, the level of inactivation obtained varies from 0.64 to 4.94 log
cycles. These results support that establishing the treatment conditions for any
industrial specic application will require validating the process with the microorganisms of concern. Although, the potential of the PEF technology for inactivating
microorganisms in must and wine at lower temperatures than those required in
thermal processing has been demonstrated, studies on the effect of the PEF treatments on the sensory properties of wine should be conducted before application of
the technology in the wineries.
The improvement of fermentation by application of PEF treatments at low electric
eld strength (0.1 and 6 kV/cm) to the yeast population before inoculation is other
potential application of the technology in wineries. A positive impact of PEF
treatment of S. cerevisiae yeast on the fermentation process of wine has been
observed in terms of mass losses, consumption of sugars, and synthesis of proteins.
Reduction of fermentation time may be of interest in the wineries in order to avoid
growth of undesirable microorganisms and to improve its productive capacity.

Conclusions
One of the most investigated application of PEF technology in the last years has been
the use of this technology for improving different operations conducted in wineries.
Research conducted has demonstrated that a pretreatment of grapes by PEF may
increase the must expression yield, reduce the maceration time during red
winemaking, and increase the color and concentration of polyphenolic compounds
in red wine without impairing sensory attributes. On the other hand, inactivation of
spoiling microorganisms by PEF may contribute to improve the wine quality by
assuring reproducible and correct fermentation completion or by reducing or
replacing the use of SO2. Finally, this technology may contribute to valorize winery
waste by improving extraction polyphenols to be used as food colorants of natural
origin or dietary supplements.
While some of these applications such as improving polyphenols extraction
during red winemaking have already been evaluated in wineries, the promising
results obtained for other applications at laboratory scale should be validated in
industrial tests. The low energy consumption of the PEF for the applications in
wineries, the availability of PEF generators with sufcient power to meet the
production capacity of the wineries, and the easy implementation of the treatment

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G. Saldaa et al.

Fig. 4 Integration in a winery of a PEF treatment chamber to process grapes before macerationfermentation

chambers into the existing processing lines of wineries should contribute to become
PEF in a viable technology in the wineries (Fig. 4).
Acknowledgment This work has been supported by the European Commission (635632FieldFOOD-H2020-SFS-17-2014).

Cross-References
Application of Pulsed Electric Energy for Grape Wastes Biorenery
Basic Concepts of High Voltage Pulse Generation
Electric Field Distribution and Electroporation Threshold
Electroporation and Electropermeabilization
Energy and Cost Analyses of Pulsed Electric Field Applications
High-Voltage Electrical Discharge Assisted Extraction of Phenolic Compounds
from Grape Seeds
Impact of Pulsed Electric Field Treatment on Must and Wine Quality
Industrial Pulsed Electric Fields Systems
Large-Scale Pulsed Electric Field Treatment Devices: Overview of Commercially
Available Equipment
Optimization of Pulsed Electric Field Treatment Chamber
Process Design, Improvement and Integration of Pulsed Electric Field
Pulsed Electric Field Treatment for Beverage Production and Preservation
Pulsed electric eld treatment for fruit and vegetable processing
Pulsed Electric Fields Assisted Extraction of Polyphenols from Grape Pomace

Pulsed Electric Fields in Wineries: Potential Applications

17

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