Trends in Food Science & Technology 20 (2009) 544e556
Review
Effects of pulsed
electric fields on
bioactive compounds
in foods: a review
Robert Soliva-Fortunya,
Ana Balasab, Dietrich Knorrb
and Olga Martı́n-Bellosoa,*
a
Department of Food Technology, TPV-XaRTA,
University of Lleida, Rovira Roure 191, 25198 Lleida,
Spain (Tel.: D34 973 702 593; fax: D34 973 702 596;
e-mail: omartin@tecal.udl.es)
b
Department of Food Biotechnology and Process
Engineering, Berlin University of Technology, Königin-
Luise-Str. 22, D-14195, Berlin, Germany
Pulsed electric fields have the potential to be used as alterna-
tive to other conventional techniques of food production. In
the last few years, several studies have demonstrated the abil-
ity of intense treatments to obtain safe and shelf-stable liquid
foods. On the other hand, novel applications such as improve-
ment of mass transfer processes or generation of bioactive
compounds by using moderate field strengths are under cur-
rent development. However, the effects of pulsed electric field
treatments on minor constituents of foods, namely on bioac-
tive compounds, have not been always considered. This
review aims at updating the state of the art regarding the
impact of pulsed electric field processing conditions on the
stability of bioactive compounds in food systems.
Introduction
The development of pulsed electric field (PEF) technol-
ogy for non-thermal food processing applications has been
one of the major fields of research of the last decade regard-
ing the study of alternative energy efficient applications.
* Corresponding author.
0924-2244/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tifs.2009.07.003
PEF technology consists in the delivery of short high
power electrical pulses (ms or ms) to a product placed in
a treatment chamber confined between electrodes. Typical
systems for the treatment of pumpable fluids consist of
a PEF generation unit, which is in turn composed of
a high voltage generator and a pulse generator, a treatment
chamber, a suitable product handling system and a set of
monitoring and controlling devices (Fig. 1).
The permeabilising effect on biological membranes that
results from the application of pulsed alternating electrical
currents is known from the beginning of the 20th century,
but it was during the 1990s that thorough research on this
technology was undertaken. On the one hand, most studies
on high-intensity treatments have been devoted to the de-
sign of pasteurization applications for liquid products, in
order to attain proper microbial destruction levels as well
as to observe an impact on enzyme activity. On the other
hand, treatments of mild or moderate intensity are currently
under study to prospect their potential of permeabilising tis-
sue structures, thus allowing the implementation in the con-
text of already existing operations such as extraction or
drying. Furthermore, PEF can also provide a potential to
induce stress reactions in plant systems or cell cultures, so
that bioproduction of certain compounds can be enhanced
or stimulated (Toepfl, Mathys, Heinz, & Knorr, 2006).
The increasing consumer interest for high nutritious
fresh-like food products, together with the search for envi-
ronmentally friendly processing technologies, has aided in
the development of emerging non-thermal technologies
such as pulsed electric fields. However, little has been pub-
lished about the effects of PEF treatments on the bioactive
compounds in foods. Tentative studies suggest that PEF
treatments can be a feasible way of assuring pasteurization
treatments without substantially affecting the nutritional
composition of food products. In this review, an update
on the state of the art on PEF technology is presented,
with special focus on the mechanisms that rule both the
generation of endogenously present bioactive compounds
at low or moderate treatment intensities and the presence
of these health-related compounds when high intensities
are applied.
Mechanism of electropermeabilisation
The application of an external electric field on biological
cells (animal, plant or microbial) results in disruption of the
cell membrane. Till date a number of theoretical models
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Fig. 1. Schematics of a PEF processing system for pumpable products.
have been suggested, but there is still no clear evidence on
the underlying mechanism of action at the cellular level.
The most accepted theory is the electromechanical model
introduced from Zimmerman, Pilwat, and Riemann
(1974). It considers the cell membrane to be a capacitor
with a low dielectric constant. Free charges of opposite po-
larities are present at both sides of a membrane (internal
and external), resulting in a naturally occurring transmem-
brane potential. Exposure to an electric field induces accu-
mulation of cell interior and exterior charges across the
membrane and therefore an increase in the transmembrane
potential. Additional potential induced from external elec-
tric field raises compression pressure on the membrane
due to attraction between opposite charges on both sides
of the membrane simultaneously which induces membrane
thinning. Electrostatic attraction between the two sides of
a membrane is further increased due to thinning of a mem-
brane. When electrocompression exceeds the elastic resis-
tance of a membrane locale rupture with pore formation
occurs in the sub-microsecond range at a given value of
field applied.
Pore formation is a dynamic process and can be revers-
ible or irreversible depending on the treatment intensity.
When pores induced are small in comparison to the mem-
brane area and are generated with PEF treatment of low in-
tensities the electric breakdown is reversible (Angersbach,
Heinz, & Knorr, 2000). The viability of the cell is main-
tained and additional biosynthesis of secondary metabolites
can be triggered as a response to the stress condition in-
duced from PEF treatment. Increasing treatment intensity,
by increasing electric field strength and/or treatment time
545
(which considers number of pulses and pulse width applied
in a batch system) will promote formation of large pores
and reversible permeabilisation will turn into irreversible
breakdown. Critical electric field strength to induce mem-
brane permeabilisation is dependent on cell geometry and
size, in the range of 1e2 kV/cm for plant cells (cell size
40e200 mm) and in the range of 12e20 kV/cm for micro-

organisms (cell size 1e10 mm) (Heinz, Alvarez, Anger-
sbach, & Knorr, 2002).
Based on this phenomenon, electroporation (permeabili-
sation of the cell membrane caused by external electric
field) was studied in practical applications on various bio-
logical systems in the field of medicine and biology. Al-
though recently the use of moderate field strength is
considered in food science, most of the studies were done
in the range of irreversible membrane permeabilisation,
with the main objective to induce microbial inactivation
or to facilitate extraction of specific constituents and/or to
increase drying rates. Application of moderate electric
fields in food technology attracted large attention in the
last decade, but still very little information is available re-
garding membrane permeabilisation kinetics and revers-
ible-irreversible structure changes of cells in real food
systems during and after application of PEF.
Impact of mild/moderate PEF treatment on bioactive
compounds in foods
The application of Pulsed Electric Field (PEF) technol-
ogy in food processing has attracted large interest in the
last couple of decades not only as a new non-thermal pres-
ervation technology, but it offers a number of other useful
546 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
applications in the food industry. Improvement of intracel-
lular metabolite extraction (Ade-Omowaye, Angersbach,
Eshtiaghi, & Knorr, 2001; Fincan, DeVito, & Dejmek, 2004;
Geulen, Teichgraeber, & Knorr, 1994; Tedjo, Eshtiaghi,
& Knorr, 2002), enhancement of drying efficiency (Ade-
Omowaye et al., 2001; Taiwo, Angersbach, & Knorr,
2002), modification of enzymatic activity (Giner, Rauret-
Arino, Barbosa-Canovas, & Martin-Belloso, 1997; Yoem,
Streaker, Zhang, & Min, 2000), preservation of certain
food ingredients (Jia, Zhang, & Min, 1999) and production
of secondary metabolites by inducing stress reactions in
plant systems (Balasa, Toepfl, & Knorr, 2006; Guderjan,
Toepfl, Angersbach, & Knorr, 2005) and cell cultures (Dör-
nenburg & Knorr, 1993) are being investigated.
However, information available concerning the effect of
Moderate Electric Fields (MEF) on bioactive components is
more limited compared to results on food pasteurization
with high-intensity PEF treatments, and the precise mecha-
nism of PEF-induced changes is still not understood in
depth.
The application of low intensity treatment does not nec-
essarily cause irreversible cell rupture. This physical pro-
cess (reversible membrane permeabilisation) was
investigated by Angesbach, Heinz, and Knorr (2000).
They reported formation of pores across the cell membrane
when a transmembrane potential of 1.7 V was reached (po-
tato tissue). Conductive channels that occurred across the
membrane resealed again within very short time and the
cell membrane recovered its electrically insulating proper-
ties. Such a phenomenon offers numerous possibilities to
induce stress reactions in plant systems or cell cultures,
whereas cells regain their vitality and metabolic activity.
Table 1 shows some applications of PEF treatments im-
proving the recovery of metabolites of interest from differ-
ent food matrices. Dörnenburg and Knorr (1993)
investigated the influence of low intensity PEF treatment
(0e30 pulses of 0e1.6 kV/cm) on production and recovery
of secondary metabolites from cultured plant tissues (Che-
nopodium rubrum and Morinda citrifolia) in correlation
with cell vitality. A very high amount of the total amaran-
thin content (85%) from C. rubrum and only 5.7% of total
amount of anthraquinones from M. citrifolia was released
after PEF treatment (10 pulses of 1.6 kV/cm and 1 pulse
at 0.75 kV/cm, respectively). Regarding these two plant va-
rieties, electroporation had noticeable different effect on
pigment release. Results obtained from PEF-treated C. ru-
brum revealed larger impact of electric field intensity on
pigment release than number of pulses at a given voltage.
Increasing the field strength, substantial pigment release
was achieved before cell viability was lost. However, treat-
ment of M. citrifolia failed to result in satisfying pigment
release at 0.5 kV/cm and 3 pulses when cell viability was
already lost. Hence, this study indicates that the potential
of effectiveness of permeabilisation procedure depends on
the location of the metabolites within the cell (cytoplasm
or vacuoles). Unfortunately, there are no other studies
available till date on the same topic (to our best knowl-
edge), or they are unsuitable for comparison due to differ-
ent treatment conditions and/or different PEF equipment
design.
Ye, Huang, Chen, and Zhong (2004) investigated electric
field stimulation of secondary metabolites production from
suspension cultures of Taxus chinensis, whereas the exper-
imental design does not correspond to the definition of what
is considered as PEF treatment within this article. The elec-
trodes of the PEF system used for the study were not im-
mersed in the treated solution. Therefore the noticed
increase in intracellular accumulation of taxuyunnanine (a
bioactive secondary metabolite) was most likely stimulated
by electro-magnetic field induced from such high electric
field intensity (60 kV and electrode distance of 6 cm).
PEF-induced recovery of plant oils with simultaneous
release of intracellular pigments and additional production
of secondary metabolites in plant systems (maize, olives
and soybeans) was studied by Guderjan et al. (2005).
They did not only report higher oil yield obtained from
maize germ, fresh olives and soybeans, but increased phy-
tosterol production in maize germs (up to 32.4%) was no-
ticed together with increased yield of isoflavonoids in
soybeans (20e21%), after application of PEF treatment
with field strengths in a range of reversible cell permeabi-
lisation (0.6 to 1.3 kV/cm). Interestingly, such a high in-
crease of phytosterols and oil yield from maize germ
could not be asserted by irreversible membrane permeabi-
lisation at 7.3 kV/cm and 120 pulses. This shows that the
application of a low PEF treatment provides a potential
to induce stress reactions in plant systems by stimulating
metabolic activity and accumulating secondary
metabolites.
Secondary metabolite production together with extract-
ability of plant pigments from wine grapes (Vitis Vinifera)
induced by low intensity pulsed electric fields was investi-
gated by Balasa et al. (2006). Increase of total polyphenolic
(TP) content (13e28%) in fresh pressed grape juice ob-
tained from PEF-treated grapes (0.5 to 2.4 kV/cm and
50 pulses) was observed. With increasing treatment inten-
sity by increasing electric field strength, a higher content
of total polyphenolics was noticed as well as higher degree
of cell membrane permeabilisation. Interestingly, different
results were obtained concerning the extractability (follow-
ing aqueous extraction) of total polyphenolics from grape
marc. After applying an electric field strength of 0.5 kV/
cm (specific energy input of 0.1 kJ/kg) increase of TP con-
tent was much higher (24%) than with PEF treated grapes
at 2.4 kV/cm (specific energy input of 2.3 kJ/kg) where
TP content resulted in increase of 14% (comparison to un-
treated sample). It was suggested by the authors that the
reason for such an increase at low electric field intensities
could have been stress induction in grape cells and subse-
quent production of polyphenolics as secondary metabo-
lites. Additionally, no change in pH, brix-values and wine
acids was noticed after PEF treatment.
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556 547

Table 1. Effect of high-intensity pulsed electric field treatments on metabolite recovery from different sources

Compound Product Treatment conditions Effects Reference

Amaranthin Cultured plant 0e1.6 kV/cm, Metabolite recovery. Increased pigment release with Dörnenburg and
tissue: C. rubrum 0e30 pulses higher loss of cell viability. More effective pigment Knorr (1993)
release at increasing field strength, than with
increasing number of pulses.
Antraquinones Cultured plant 0e1.6 kV/cm, Metabolite recovery. No substantial pigment release Dörnenburg and
tissue: M. citrifolia 0e30 pulses at 0.5 kV/cm and 3 pulses where cell viability was Knorr (1993)
already lost.
Oil and Maize germ 0.6 kV/cm; 0.62 kJ/kg Higher oil yield (up to 88.4%) and increased Guderjan et al.
Phytosterol 7.3 kV/cm; 91.4 kJ/kg amount of phytosterols (up to 32.4%) was reached (2005)
simultaneously. Oil yield increased marginal
by 2.9% in comparison to untreated sample, and
phytosterol increase reached 14.7%.
Oil Olives 0.7 kV/cm, 30 pulses Higher oil yield of 6.5% and 7.4% respectively. Guderjan et al.
1.3 kV/cm, 100 pulses (2005)
Isoflavonoids Soybeans 1.3 kV/cm, 50 pulses, Amount of isoflavonoid diadzein increased Guderjan et al.
1.857 kJ/kg 1.3 kV/cm, by 20% in comparison to the referent sample. (2005)
20 pulses, 0.743 kJ/kg A rise of 21% of isoflavonoid genistein in
comparison to the reference was reached.
Polyphenolics Grapes 0.5 kV/cm, 50 pulses, Increase of total polyphenolic (TP) content Balasa et al.
0.1 kJ/kg 2.4 kV/cm, in fresh pressed grape juice of 13% was reached in (2006)
50 pulses 2.3 kJ/kg comparison to referent sample simultaneously with
24% increase of TP content in grape residue. 28%
increase of TP content in fresh pressed grape juice
was reached simultaneously with 14% higher TP
content in grape residue, in comparison to
referent sample.
Juice and Grapes 3 kV/cm, 50 pulses Higher juice yield (75%) of PEF-treated vine grapes Tedjo et al.
Anthocyanins in comparison to referent sample (70%). Total (2002)
anthocyanin content was almost 3 times higher
than of untreated grapes.
Betalain Beetroot tubers 1 kV/cm, 270 Solid-liquid extraction of beetroot pigment. 90% Fincan et al.
(B. vulgaris) pulses, 7 kJ/kg of total red coloring was released (release of betalain (2004)
and ionic species), in comparison to mechanical
pressing. Increase in tissue electric conductivity after
PEF treatment correlated approximately linearly with
the extraction yield of red pigment and ionic species
(only up to extraction level of 60e80%).
Sucrose Sugar beet 1.2e2.5 kV/cm, Sucrose extraction at ambient temperature Eshtiaghi and
1e200 pulses after PEF application for tissue disintegration. Knorr (2002)
Higher degree of sucrose extraction has been
obtained, higher dry matter (w30%) in pressed
pulp, shorter time of extraction.
Juice and Carrots (La rosa 0.6e2.6 kV/cm, Juice yield increased from 30.1% of untreated Geulen et al.
Carotenoids di Chioggia) 5e100 pulses material to 76.1% of PEF-treated (2.6 kV/cm, 50 pulses). (1994)
50 ms; 1 Hz No differences were detected in pH value,
total acids and dry substances after PEF treatment
in comparison to reference. Carotenoid content
(ß-carotene) has been maintained.
Vitamin C Bell peppers 0.5e2.5 kV/cm, Osmotic dehydratation (OD) was enhanced Ade-Omowaye
Carotenoids 20 pulses 400 ms, after application of PEF pre-treatment. et al. (2002)
2 Hz The percentage of vitamin C reduction
after PEF treatment prior to OD was lower
than pre-treated at different temperatures (from 25
to 55  C). Carotenoid reduction as a result of temperature
increase was z 80% to 55%, and z74% to 62% of
initial fresh content as a result of PEF pre-treatment.
Vitamin C Red bell 2 kV/cm, 1e50 pulses Vitamin C retention after osmotic dehydration / Ade-Omowaye
peppers untreated > frozen > PEF-pretreated samples with et al. (2003)
1 > 5 > 50 > 10 > 20 pulses. The data provided
clearly indicate improved mass transfer during
drying as a consequence of PEF pre-treatment and
also suggested minimal loss of vitamin C as quality
indicator due to electric field treatment.
(continued on next page)
548 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Table 1 (continued )
Compound Product Treatment conditions
Oil Tocopherols Rapeseed 5 kV/cm, 60 pulses
Polyphenols 7 kV/cm, 120 pulses
Apple (Roter 1, 3 5 kV/cm, 30 pulses
Boskoop) untreated control
juice and juice after
pectolytic mash treatment of 5 kV/cm. Antioxidant capacities (TEAC, FRAP,
PEF-induced cell permeabilisation and release of intra-
cellular pigments (anthocyanins) from wine grapes was
studied as well by Tedjo et al. (2002). After applying an
electric field strength of 3 kV/cm and 50 pulses, 65% of
the total membrane area was permeabilised, which resulted
in threefold increase of the total anthocyanin content (as
compared to untreated sample). With resemblance to
work done by Balasa et al. (2006), it could be again no-
ticed that the higher the electric field strength, the higher
the degree of cell membrane permeabilisation and subse-
quent release of intracellular compounds. In agreement
with this theory, Eshtiaghi and Knorr (2002) showed com-
pletely sucrose depleted sugar beet samples (relative gain
of approx. 98%) when applying pulsed electric field (20
pulses of 2.5 kV/cm) as pre-treatment to a conventional
extraction. Furthermore, it was shown that PEF-pretreated
beets can easily be pressed and higher yield of sucrose ob-
tained, that the pressed pulp from PEF pre-treated samples
in comparison to the conventional thermal process contain
less residual sugar, and that shorter extraction times are
required.
Fincan et al. (2004) investigated extractability of beet-
root pigment (betalain, the main constituents of red color-
ing of beetroots, accumulates in vacuoles) and different
ionic species from fresh red beetroot tubers (Beta vulgaris),
using low intensity PEF treatments. 90% of total red color-
ing was released into the isotonic solution (following 1 h
aqueous extraction) after applying 270 rectangular pulses
of 10 ms at field strength of 1 kV/cm. The amount of ex-
tracted pigment was found to be directly proportional to
the release of ionic species, thus authors postulated that
no differential permeabilisation of any of the intracellular
compartments occurred.
Impact of low intensity PEF treatments (0.6 to 2.6 kV/
cm, 5e100 pulses with frequency of 1 Hz) on b-carotene
content of carrot juice obtained from PEF-treated carrots
was studied by Geulen et al. (1994). When applying PEF
treatment with a field strength of 2.6 kV/cm and 50 pulses,
juice yield increased from 30.1% (untreated sample) to
76.1%, while no significant difference was noticed in b-car-
otene content, pH value, dry substances and total acids.
Effects Reference
Increased content of polyphenols and tocopherols, Guderjan et al.
which are important antioxidants in rapeseed, most (2007)
likely caused increased antioxidant capacity, measured
in PEF-treated samples.
Polyphenolic content of PEF-treated samples Schilling et al.
with 1 and 3 kV/cm were insignificant, and slightly (2007)
lower contents were partially observed after treatment
DPPH) remained unchanged
Considering pH, TSS, TA and the sugar-acid ratio, the
juices obtained after PEF treatment of apple mash did
not differ from the respective controls.
The effectiveness of PEF treatment (from 0.5 to 2.5 kV/cm)
on b-carotene and vitamin C content in bell peppers, as
a pre-treatment to osmotic dehydration (OD) in comparison
to elevated temperature treatment (from 25 to 55  C) was
investigated by Ade-Omowaye, Rastogi, Angersbach, and
Knorr (2002). A higher vitamin C content (182.8 mg/
100 g ids) was determined when an electric field strength
of 2.5 kV/cm (20 pulses, pulse duration 400 ms, frequency
of 2 Hz) was used than in osmosed bell pepper at 55  C
(55.7 mg/100 g ids). Additionally, lower reduction of b-
carotene content was observed after PEF treatment in com-
parison to b-carotene reduction after treatment at elevated
temperature. The same research group (Ade-Omowaye,
Taiwo, Eshtiaghi, Angersbach, & Knorr, 2003) showed
that vitamin C retention immediately after PEF treatment
(50 pulses at 2 kV/cm, pulse duration 400 ms) ranged
from 89.6 to 96.5%, from which it could be concluded
that PEF itself has no detrimental impact on vitamin C.
Another application of pulsed electric fields as a pre-treat-
ment method before mechanical pressing or solvent extrac-
tion of rapeseed and its impact on functional food
ingredients (antioxidants, tocopherols, polyphenolics and
phytosterols) was studied by Guderjan, Elez-Martı́nez, and
Knorr (2007). After applying PEF treatments (60 pulses of
5 kV/cm, and 120 pulses of 7 kV/cm; pulse duration
30 ms), oil yield increased by pressing as well as with solvent
extraction and additionally higher concentrations of tocoph-
erols, polyphenols and phytosterols were measured in oil.
The total polyphenolic content of PEF-treated samples
(120 pulses at 7 kV/cm) after pressing was three times higher
than of non-treated rapeseed. With increased PEF treatment
intensity, by increasing field strength and number of pulses,
higher amounts of tocopherols and polyphenols in oil were
observed. The higher content of polyphenols and tocoph-
erols, which are important antioxidants in rapeseed, most
likely caused increased antioxidant capacity measured in
samples treated with high-intensity electric field pulses.
The antioxidative capacity of juice obtained from PEF-
treated apple mash was investigated by Schilling et al.
(2007), where no significant differences were found be-
tween the control groups and samples treated with electric
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
field intensities of 1, 3 and 5 kV/cm, and 30 pulses. In-
creased yield of valuable ingredients using PEF treatment
was not observed as reported for some other fruits, but it
stayed unchanged for samples treated with electric field
strengths of 1 and 3 kV/cm, and a slightly lower content
of apple polyphenolics was partially noticed after a treat-
ment of 5 kV/cm.
In summary, many reports have demonstrated advan-
tages of the application of moderate electric field pulse
technology, including higher juice yield as well as high
juice purity, and unchanged or upgraded antioxidant capa-
city of the juice in combination with low energy consump-
tion. Good pressing efficiency, faster extraction processes
and improvement in drying processes are shown as advan-
tages to standard (conventional) treatments. The observed
effects are difficult to compare due to different electric field
systems used, different electric process parameters, insuffi-
ciently described equipment and/or treatment conditions.
These parameters should be standardized to enhance the
evaluation of the processes described. There are several
important parameters that should be concerned: electric
field strength, E (kV/cm); total treatment time; number of
pulses; pulse geometry; treatment chamber design; flow
rate of the product (for continuous systems); fluctuation
of temperature; total energy input.
Effects of high-intensity PEF treatments on bioactive
compounds in foods
Proteins, peptides and amino acids
Little detailed work has been conducted to date on the
effects of PEF on food proteins. According to Chen and
Wu (2006) the possible mechanisms involved in the loss
of functionality include Joule heating damages, electrocon-
formational changes, or both. From the point of view of
food preservation, it has been demonstrated that treatments
with pulse widths of a few microseconds can attain micro-
bial inactivation without significant damage in the protein
matrix (Perez & Pilosof, 2004). However, PEF treatments
can also be used to induce changes in the protein structure
and functionality. Most research works studying the effect
of PEF on proteins have been carried out on egg white
and whey proteins, probably because of their good perfor-
mance as gelling agents in many food products.
Among as many as 40 different proteins contained in the
egg white, the major proteins involved in the heat-induced
gelation are ovalbumin (54%), conalbumin (12%), ovomu-
coide (11%) and lysozyme (3.5%). Ovalbumin is the most
heat-stable whereas conalbumin is the most heat-sensitive
(Fernandez-Diaz, Barsotti, Dumay, & Cheftel, 2000). Mar-
tı́n-Belloso et al. (1997) reported no protein coagulation
when liquid whole egg was exposed to up to 100 square-
wave pulses of 2e4 ms using an electric field intensity of
26 kV/cm. The application of 10 pulses per unit volume
of exponential decay pulses of 2 ms with an electric field
of 48 kV/cm did not alter the electrophoretic pattern of
egg proteins (Ma, Chang, Góngora-Nieto, Barbosa-
549
Cánovas, & Swanson, 2001). Consistently, surface hydro-
phobicity of egg white proteins was not increased by apply-
ing from 2 to 8 exponential pulses of 20e35 kV/cm at
100e900 Hz (Jeantet, Baron, Nau, Roignant, & Brulé,
1999). Barsotti, Dumay, Mu, Fernandez-Diaz, and Cheftel
(2002) reported that a series of 20 exponential decay pulses
of 31.5 kV/cm at 1 Hz applied to ovalbumin in a buffer so-
lution induced at least a partial unfolding of the protein
structure or enhanced the ionization of SH groups into
a more reactive S form. The number of pulses and the en-
ergy applied per pulse significantly influenced SH group re-
activity. However, these modifications appeared to be
transient, provided that when the pulse-processed
ovalbumin solution was kept at 4  C for more than
30 min the extent of the observed changes decreased and
were completely reversed after 24 h.
b-Lactoglobulin is the primary gelling agent in whey,
undergoing complex changes upon heating (Perez & Pilo-
sof, 2004). Barsotti et al. (2002) observed that the fourth
derivative of the UV spectra of native and pulse-processed
b-lactoglobulin solutions were identical and, in turn,
differed significantly from the spectra of heated solutions,
which exhibited shifting peaks for tyrosine and tryptophan,
and therefore concluded that PEF treatments (31.5 kV/cm,
200 exponential decay pulses applied at 1 Hz, 2.3 J/pul-
se mL) shown to cause a 2e5 log cycle inactivation of
E. coli NCTC 9001 did not induce marked unfolding of
the b-lactoglobulin molecule. In contrast, Perez and Pilosof
(2004) reported a partial modification of up to 40% of the
native structure of b-lactoglobulin using 10 exponentially
decaying pulses of 2 ms at 12.5 kV/cm. These results
were supported by electrophoresis analyses, indicating for-
mation of aggregates involving covalent bonds. It was also
shown that the thermal stability of b-lactoglobulin was
greatly reduced when applying PEF treatments, in contrast
with a relative enhancement of the heat resistance of egg
white proteins. Odriozola-Serrano, Bendicho-Porta, and
Martı́n-Belloso (2006) found significant differences be-
tween PEF-treated and thermally treated milk for a-lacto-
globulin, b-lactoglobulin and serum albumin. The effect
of PEF treatments on other milk proteins has been scarcely
studied. No detectable changes in the secondary structure
or the thermal stability of secondary structure were detected
by Li, Bomser, and Zhang (2005) with a PEF bipolar treat-
ment at 41.1 kV/cm for 54 ms, contrasting with the signifi-
cant alteration of the secondary structure and loss of
immunoactivity after heat treatments.
Very little is known about the effect of PEF on protein
functionality. In the case of soybean, a protein with high
biological value, Li and Chen (2006) reported a significant
increase in both solubility, emulsibility, foaming capacity,
hydrophobicity and degree of denaturation of a protein iso-
late with increasing electric field (up to 41.1 kV/cm) and
treatment time (up to 91 ms).
On the other hand, proteins with enzymatic activity can
be inactivated under some PEF processing conditions
550 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556

(Martı́n-Belloso & Elez-Martı́nez, 2005; Yeom & Zhang,
2001). Results from several studies indicate that field
strength, pulse width, number of pulses and pulse shape
are the main critical parameters affecting enzyme inactiva-
tion. The influence of food matrix, pH and temperature
should not be neglected. Several authors described temper-
ature distribution inside PEF treatment chamber (Fiala,
Wouters, van den Bosch, & Creyghton, 2001; Gerlach
et al., 2008; Lindgren, Aronsson, Galt, & Ohlsson, 2002).
Critical zones which are located near the insulators may
reach temperatures of 90  C which could have a detrimental
impact on either enzymes or any heat-labile compound.
Unfortunately this temperature inside the treatment cham-
ber cannot be measured, but it can be detected through
mathematical modelling of such processes. Enzymes are
as well affected by changes in pH. Due to electrochemical
reactions that might occur at the electrode surface (Morren,
Roodenburg, & Han, 2003) partial electrolysis of solution
can take place and therefore a local pH change may cause
changes in enzyme activity. Nevertheless, the mechanism
of enzyme inactivation is still unclear. Contrariwise to the
extensive research conducted on microorganism inactiva-
tion, PEF studies on enzymes are not as systematic and of-
ten show controversial results (Table 2). Some enzymes are
strongly inactivated, such as polyphenol oxidase (PPO) (Gi-
ner et al., 2000; Ho, Mittal, & Cross, 1997; Yang, Li, &
Zhang, 2004a; Zhong et al., 2007) pectin methyl esterase
(Elez-Martı́nez, Suárez-Recio, & Martı́n-Belloso, 2007; Es-
pachs-Barroso, van Loey, Hendrickx, & Martı́n-Belloso,
2006; Giner, Gimeno, Barbosa-Cánovas, & Martı́n, 2001;
Giner et al., 2002; Rodrigo, Barbosa-Cánovas, Martı́nez,
& Rodrigo, 2003; Yeom, Streaker, Zhang, & Min, 2000),
papain (Yeom, Zhang, & Dunne, 1999), plasmin (Vega-
Mercado, Powers, Barbosa-Cánovas, & Swanson, 1995), li-
pase (Bendicho, Estela, Giner, Barbosa-Cánovas, & Martı́n,
2002; Ho et al., 1997), protease (Bendicho, Barbosa-
Cánovas, & Martı́n, 2003a), glucose oxidase, lysoenzyme,
amylase (Ho et al., 1997) or alkaline phosphatase (Castro,
Swanson, Barbosa-Cánovas, & Zhang, 2001). However,
other researchers have reported no significant influence of
PEF or even an increase in enzyme activity. It is generally
known that pulses of increasing width cause a more severe
modification of enzyme structure. For instance, almost no
inactivation of alkaline phosphatase in milk was observed
after 20 pulses of 0.39 ms at 21.5 kV/cm (Grahl & Märkl,
1996), whereas 65% inactivation was obtained with 70
pulses of 0.7 ms at 22 kV/cm (Castro et al., 2001). Some
authors have suggested that conformational changes in en-
zyme structure may be responsible for the modifications in
enzyme activity due to PEF treatments. Yeom et al. (1999)
related the reduction of papain activity to the loss of a-helix
structure. Giner et al. (2002) attributed the differences in
the inactivation of PPO and PME by PEF to the differences
in the molecular enzyme structures. Consistently, Yang, Li,
and Zhang (2004b) found that inactivation of pepsin by
PEF was correlated to the loss of b-sheet secondary struc-
ture of pepsin molecules. Similar results were reported by
Zhong et al. (2007) who reported dramatic changes in the
secondary structure of PPO and peroxidase (POD) enzymes
treated with intense treatments of 25 kV/cm for 1740 ms
and 744 ms, respectively. Detailed studies are in progress
to possibly identify the reasons for controversial results
concerning enzyme activity after PEF treatments.
There is no evidence that PEF treatments affect the
primary structure of proteins (peptides and amino acids).
Results obtained by Garde-Cerdán, Arias-Gil, Marsellés-
Fontanet, Ancı́n-Azpilicueta, and Martı́n-Belloso (2007)
support this statement, thus indicating that intense treat-
ments, consisting of 4-ms pulses of 35 kV/cm applied at
1000 Hz for an effective treatment time of 1 ms, did not
induce substantial changes in the amino acids profile and
overall content of grape must.

Table 2. Inactivation of enzymes in foods by high-intensity pulsed electric field treatments

Enzyme Media Treatment intensity Inactivation (%) Reference

Polyphenoloxidase Apple juice 31 kV/cm, 1000 ms <10 Van Loey, Verachtert, and Hendrickxn (2002)
Peroxidase Milk 21.5 kV/cm, 20 pulses 25 Grahl and Märkl (1996)
19 kV/cm, 500 ms 0 Van Loey et al. (2002)
Orange juice 35 kV/cm, 1500 ms, 35  C 100 Elez-Martı́nez, Aguiló-Aguayo, and
Martı́n-Belloso (2006)
‘Horchata’ 25 kV/cm, 300 ms, 35  C 72.4 Cortés et al. (2006)
Lipoxygenase Green pea juice 20 kV/cm, 400 ms 0 Van Loey et al. (2002)
Tomato juice 35 kV/cm, 50 ms, 30  C 80 Min, Min, and Zhang (2003)
Pectin methyl Orange juice 35 kV/cm, 59 ms, 60.1  C 88 Yeom, Streaker, and Zhang (2000)
esterase 35 kV/cm, 1500 ms, 35  C 90 Yeom, Zhang, and Chism (2002)
35 kV/cm, 1500 ms, 35  C 80 Elez-Martı́nez et al. (2007)
Orange-carrot 35 kV/cm, 1500 ms, 35  C 81.4 Rodrigo et al. (2003)
juice blend
Protease Skim milk 14e15 kV/cm, 196 ms 60 Vega-Mercado, Powers, Barbosa-Cánovas,
Luedecke, and Swanson (2001)
35.5 kV/cm, 866 ms, 46  C 81.1 Bendicho et al. (2003a)
Whole milk 35.5 kV/cm, 866 ms, 46  C 57.1 Bendicho, Barbosa-Cánovas, and Martı́n (2003b)
Lipase Milk 21.5 kV/cm, 20 pulses 60 Grahl and Märkl (1996)
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Phenolic compounds
Little information is available about the effect of PEF
treatments on complex phenolic molecules, on the phenolic
content and composition of foods and with possible interac-
tions with other compounds found in the food matrix.
Since some of these compounds are partially responsible
for the colour of some fruits, colour determinations can be
a good indicative of the effect of PEF treatments on the
quantitative content these compounds, especially in those
food products containing high amounts of anthocyanins.
No significant colour modifications have been found be-
tween non-treated samples and a cranberry juice treated
with 40 kV/cm for 150 ms (Jin & Zhang, 1999) or a yo-
gurt-based drink containing a strawberry fruit syrup (Evren-
dilek, Yeom, Jin, & Zhang, 2004), indicating that the
changes induced by PEF treatments appear to be scarce.
In addition, little has been published on the effect of PEF
on the qualitative phenolic content. Sánchez-Moreno et al.
(2005) evaluated the effect of a PEF treatment at 35 kV/cm
for 750 ms with 4-ms bipolar pulses at 800 Hz on the flava-
none content of orange juice. No changes in the total flava-
nones were observed, nor in the individual flavanone
glycosides and their aglycons hesperetin and naringenin.
Isoprenoid compounds
It is well known that carotenoids are widespread pig-
ments in a great number of plant-based foods. Nevertheless,
there is no evidence about the extent of the changes induced
by PEF treatments in their structure or bioactivity. Table 3
presents a summary of studies on the effect of PEF treat-
ments on carotenoids, as well as on other compounds
such as phenolic substances and vitamins. Cortés, Esteve,
Rodrigo, Torregrosa, and Frı́gola (2006) reported a non-sig-
nificant decrease in orange juice total carotenoids when ap-
plying bipolar treatments of 25e40 kV/cm for 30e340 ms.
Individual carotenoids with antioxidant activity (b-caro-
tene, b-cryptoxanthin, zeaxanthin and lutein) appeared to
be resistant to the applied treatments, thus resulting into
a better preservation of the antioxidant activity of the juices
as compared to the thermally pasteurized. Consistently, im-
proved stability of these compounds was achieved during
the storage shelf-life of orange juice processed under sim-
ilar conditions (Cortés, Torregrosa, Esteve, & Frı́gola,
2006). Furthermore, controversial results were reported
by Torregrosa, Cortés, Esteve, and Frı́gola (2005), who
observed that carotenoids with provitamin A effect
(a-carotene, b-carotene, z-carotene, a-cryptoxanthin and
b-cryptoxanthin) of an orange:carrot juice mixture (80:20
v/v) were affected by PEF treatments. The differences re-
ported could also be due to the different temperatures
reached during processing, given that temperatures above
65  C were reported to be achieved with treatments of
long duration.
On the other hand, studies on tomato indicate that there
is no significant difference in the concentrations of lyco-
pene between tomato juice processed by a commercial
551
scale PEF system at 40 kV/cm for 57 ms and tomato juice
thermally processed at 92  C for 90 s (Min, Jin, & Zhang,
2003). Changes in lycopene concentrations appeared to
be well correlated with colorimetric indexes, although
some differences were detected probably because of in-
creased rates of non-enzymatic browning reaction in the
thermally processed juice (Min & Zhang, 2003). As well
as for lycopene in tomato juice, spectrometric assays can
be a good indicator of the changes in the carotenoid content
of some juices. Results reported by different authors show
that colour in citrus juices was not substantially affected by
PEF treatments, indicating that no significant losses in the
total amount of carotenoids were induced (Ayhan, Yeom,
Zhang, & Min, 2001; Cserhalmi, Sass-Kiss, Tóth-Markus,
& Lechner, 2006; Elez-Martı́nez, Soliva-Fortuny, &
Martı́n-Belloso, 2006; Yeom et al. 2000). However, little
information has been published on the effect of PEF on in-
dividual carotenoid compounds in most food matrixes.
Furthermore, to our best knowledge, there is no informa-
tion available on the effect of PEF treatments on phytos-
terols and other terpenoid compounds.
Chlorophylls
Dietary chlorophyll derivatives other than naturally
abundant Mg-chlorophylls, including metallo-chlorophyl-
lins and metal-free pheophorbides, pheophytins, and pyro-
pheophytins have been identified as potential
chemopreventative agents with antioxidant and antimuta-
genic activities (Ferruzzi, Böhm, Courtney, & Schwarz,
2002). However, little is known about the effect of PEF
on these compounds. Yin, Han, and Liu (2007) proposed
the combined use of intense PEF treatments with water-sol-
uble Zn2þ concentrations to prevent chlorophyll breakdown
without significantly affecting flavour and vegetable quality
of spinach puree. These authors found that increasing field
strength, up to 60 kV/cm, increased the green colour of the
treated product, which was attributed to the destruction of
microorganisms and enzymes that mediate chlorophyll deg-
radative reactions. Field strengths higher than 60 kV/cm ap-
peared to be detrimental to the colour of spinach puree but
whether this negative effect is due to the direct action on
chlorophylls or to changes in the microstructure of the
product, thus affecting chlorophylls protection, needs still
to be investigated.
Fatty acids
High-intensity PEF treatments induce little changes in
the profile of fatty acids and their functionality. Garde-Cer-
dán et al. (2007) reported that intense treatments of 35 kV/
cm for 1 ms applied as bipolar 4-ms square pulses at
1000 Hz rendered grape juices with a similar content of to-
tal fatty acids compared to the fresh must. No changes in
individual fatty acids were found, except for lauric acid,
which seemed to be negatively affected by the action of
PEF. Consistently, no significant differences were found
in the content of free fatty acids of whole milk when
552 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556

Table 3. Effect of high-intensity pulsed electric field treatments on some health-related compounds in food systems

Compound Media Treatment intensity Effects Reference

Flavonoids Orange juice 35 kV/cm, 750 ms No changes in either individual Sánchez-Moreno et al. (2005)
flavanones nor in total content
Carotenoids Orange juice 25e40 kV/cm, 30e340 ms No significant changes in overall content. Cortés, Esteve, et al. (2006)
Better stability of individual compounds
compared to thermal pasteurization
Orange-carrot 25e40 kV/cm, 30e340 ms Rise in carotenoids content with Torregrosa et al. (2005)
juice blend increasing treatment time Increase of
compounds with provitamin A effect
at 25 and 30 kV/cm compared to
heat treatments
Tomato juice 40 kV/cm, 57 ms No changes in lycopene with Min, Jin, and Zhang (2003)
respect to thermal treatment
Vitamin B1 Milk 18e3e27.1 kV/cm, Very low or negligible reductions Bendicho, Espachs, et al. (2002)
up to 400 ms
Vitamin B2 Milk 18e3e27.1 kV/cm, Very low or negligible reductions Bendicho, Espachs, et al. (2002)
up to 400 ms
Vitamin C Protein fortified 28 kV/cm, 100e300 ms Loss increasing from 4 to Sharma et al. (1998)
orange juice-based 13% as treatment time increased
beverage
Orange juice 87 kV/cm, 40 instant Very low or negligible reductions Hodgins et al. (2002)
charge reversal pulses, 50  C
15e35 kV/cm, 100e1000 ms Loss ranging from 1.8 to 12.5% Elez-Martı́nez and
Martı́n-Belloso (2007)
Grape juice reversal pulses, 50  C Very low or negligible reductions Wu, Mittal, and Griffiths (2005)
Apple juice and cider 22e35 kV/cm, 94e166 ms Very low or negligible reductions Evrendilek et al. (2000)
‘Gazpacho’ soup 15e35 kV/cm, 100e1000 ms Loss ranging from 2.9 to 15.7% Elez-Martı́nez and
Martı́n-Belloso (2007)
Milk 22.6 kV/cm, 400 ms 6.6% depletion Bendicho, Espachs, et al. (2002)
Vitamin D Milk 18e3e27.1 kV/cm, Very low or negligible reductions Bendicho, Espachs, et al. (2002)
up to 400 ms
Vitamin E Milk 18e3e27.1 kV/cm, Very low or negligible reductions Bendicho, Espachs, et al. (2002)
up to 400 ms

applying monopolar square-wave pulses of 7 ms at a repeti-
tion rate of 111 Hz and field strength of 35.5 kV/cm during
300e1000 ms (Odriozola-Serrano et al., 2006). In accor-
dance with these results, no significant differences were
found between the contents of saturated, monounsaturated
or polyunsaturated fatty acids found in an orange juice-
milk beverage processed at 35e40 kV/cm for 40e180 ms
using 2.5-ms bipolar square-wave pulses and the contents
in untreated samples (Zulueta, Esteve, Frasquet, & Frı́gola,
2007). The slight reduction of the fat content was attributed
to a possible effect of PEF treatments on fat globule aggre-
gates by dissociating them, leading to a reduction of the sta-
bility of oil-water interface after PEF treatments.
Vitamins
High-intensity PEF treatments have been reported to
cause less change in vitamin content than conventional
processing treatments. Sharma, Zhang, and Chism
(1998) found that a protein fortified orange juice-based
beverage processed with a PEF treatment of 28 kV/cm un-
derwent vitamin C reductions of 4.0e13.0% depending on
the number of pulses applied in comparison with un-
treated juice. Consistently, very low or negligible reduc-
tions in the vitamin C content of fruit juices have been
reported when combining treatments of 20 reversal pulses
at 80 kV/cm with bacteriocin incorporation in orange
(Hodgins, Mittal, & Griffiths, 2002) and grape (Wu, Mittal
and Griffiths, 2005) juices. Qiu, Sharma, Tuhela, Jia, and
Zhang (1998) found that changes in vitamin C content of
heat-pasteurized orange juice due to processing were be-
tween 2 and 3 fold higher than those of PEF-processed
juices, which is in agreement with the results reported
by Elez-Martı́nez, Soliva-Fortuny, et al. (2006) and Evren-
dilek, Jin, Ruhlman, and Qiu (2000) in apple cider and in
a ‘gazpacho’ type cold vegetable soup.
Cortés, Torregrosa, et al. (2006) reported a decrease of
7.52% vitamin A in orange juice treated at 30 kV/cm for
100 ms, whereas a heat pasteurization treatment (90  C,
20 s) induced a loss of 15.62% of the initial vitamin A
content.
Regarding vitamins in milk, Bendicho, Espachs, Aránte-
gui, and Martı́n (2002) evaluated the effect of PEF treat-
ments on both water-soluble (riboflavin, thiamine, and
ascorbic acid) and fat-soluble vitamins (cholecalciferol
and tocopherol). No changes in the vitamin content were
observed, with the exception of ascorbic acid, which de-
creased slightly. Thus, a treatment at 22.6 kV/cm for
400 ms was more effective than either mild or intense
heat pasteurization treatments in preserving the initial vita-
min content of milk.
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Though specific conditions that assure microbial inacti-
vation have been tried in different products, there are few
studies about the effect of PEF critical parameters on vita-
mins. Elez-Martı́nez and Martı́n-Belloso (2007) reported
that pulses applied in bipolar mode, as well as decreasing
field strength, treatment time, pulse frequency and width,
led to higher levels of vitamin C retention ( p < 0.05) in
both orange juice and ‘gazpacho’ soup. PEF-treated prod-
ucts always showed vitamin C retention higher than that
of the heat-pasteurized food.
Flavour compounds
The losses of volatile compounds in PEF-treated prod-
ucts are usually dependent on the type of compound. The
amount of ethyl butyrate in orange juice decreased
a 9.7% when applying a treatment of 30 kV/cm for
480 ms at 1000 Hz with 2-ms square-wave pulses, which
contrasts with a 22.4%-depletion reached when heat
(90  C, 1 min) was used (Jia et al., 1999). In addition,
PEF treatments did not affect the amount of decanal in
the juice, which contrasts with a 41% loss that was reported
in the heat processed juice compared to the freshly
squeezed juice. In general, a loss of 3e9% was found after
PEF treatments whereas a reduction of a 22% of the volatile
compounds was reported for heat pasteurization. Min, Jin,
Min, Yeom, and Zhang (2003) and Yeom et al. (2000)
also reported a higher content of flavour components (a-pi-
nene, myrcene, octanal, limonene and decanal) in PEF-
treated orange juice (35e40 kV/cm, 59e97 ms) than in
heat-pasteurized juice (94.6  C, 30 s). Consistently, the vol-
atile aroma components of citrus juices such as orange,
lemon, grapefruit and tangerine were not significantly lost
due to PEF treatments (Cserhalmi, et al., 2006). Similar re-
sults have been reported in tomato juice treated at 40 kV/cm
for 57 ms, which retained more flavour compounds (trans-
2-hexenal, 2-isobutylthiazole, cis-3-hexanol) than ther-
mally processed or unprocessed tomato juice (Min &
Zhang, 2003). Flavour loss in the PEF-processed juices
has been frequently attributed to other factors related to
the treatment system. For instance, it has been reported
that the effectiveness towards flavour loss can be substan-
tially increased by modifying the degassing system or by
using PEF systems that allow products with a greater parti-
cle size. Nevertheless, little is know about the effects of in-
tense PEF conditions on the release of flavour compounds
from the food matrix, the mechanism and factors that
take part in the release of these compounds and, indeed,
what is the effect of PEF treatments on the flavour profile
and related compounds in different foods.
Conclusions and future research needs
Food applications of pulsed electric fields provide a po-
tential to implement energy efficient processes at an indus-
trial scale in order to obtain mildly processed products with
enhanced content of bioactive compounds. It is noteworthy
that the application of moderate pulsed electric fields
553
requires low energy inputs and may be useful to improve
the extractability of valuable bioactive compounds from
different food matrices or even induce the generation of
certain compounds when treating metabolically active tis-
sues. On the other hand, high-intensity treatments may be
applied to pasteurize liquid foods as an alternative to con-
ventional heat treatments. The application of these treat-
ments in order to obtain safe and stable products while
maintaining their fresh-like bioactive potential looks prom-
ising and requires further investigation.
Nevertheless, application of PEF technology has still sev-
eral major handicaps that need to be solved before the tech-
nology can be industrially implemented. On the one hand,
literature studies have been carried out with systems greatly
differing in both building-up, way of operation, and con-
struction materials, which makes it extreemly difficult to
compare results from different research groups. Further-
more, because of the intrinsic characteristics of the technol-
ogy, it is not easy to on-time monitor the process conditions
that determine the boundary regions delimiting areas in
which the primary treatment effects (caused by the electrical
treatment) outweigh the secondary effects (caused by heat).
Future studies should try to give response to these needs.
Studies on the effect of PEF processing parameters on the
bioactive content of treated foods need to be extensively
carried out. In-depth research is needed in order to study
the kinetics of generation, retention or degradation of
health-related compounds as affected by PEF treatment
conditions, as well as to elucidate the mechanistic insight
of the induced changes.
Acknowledgements
This study has been carried out with financial support
from the Commission of the European Communities,
Framework 6, Priority 5 ‘Food Quality and Safety’, Inte-
grated Project NovelQ FP6-CT-2006-015710.
References
Ade-Omowaye, B. I. O., Angersbach, A., Eshtiaghi, N. M., & Knorr, D.
(2001). Impact of high intensity electric field pulses on cell per-
meabilisation and as pre-processing step in coconut processing.
Innovative Food Science and Emerging Technologies, 1, 203e209.
Ade-Omowaye, B. I. O., Rastogi, N. K., Angersbach, A., & Knorr, D.
(2002). Osmotic dehydration of bell peppers: influence of high
intensity electric field pulses and elevated temperature treatment.
Journal of Food Engineering, 54, 35e43.
Ade-Omowaye, B. I. O., Taiwo, K. A., Eshtiaghi, N. M.,
Angersbach, A., & Knorr, D. (2003). Comparative evaluation of the
effects of pulsed electric field and freezing on cell membrane
permeabilisation and mass transfer during dehydration of red bell
peppers. Innovative Food Science and Emerging Technologies, 4,
177e188.
Angesbach, A., Heinz, V., & Knorr, D. (2000). Effects of pulsed electric
fields on cell membranes in real food systems. Innovative Food
Science and Emerging Technologies, 1, 135e149.
Ayhan, Z., Yeom, H. W., Zhang, H., & Min, D. B. (2001). Flavor, color,
and vitamin C retention of pulsed electric field processed orange
554 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
juice in different packaging materials. Journal of Agricultural and
Food Chemistry, 49(2), 669e674.
Balasa, A., Toepfl, S., & Knorr, D. (2006). Pulsed electric field treat-
ment of grapes. Food Factory of the Future 3, Gothenburg,
Sweden.
Barsotti, L., Dumay, E., Mu, T. H., Fernandez-Diaz, M. D., &
Cheftel, J. C. (2002). Effects of high voltage electric pulses on
protein based food constituents and structures. Trends in Food
Science and Technology, 12, 136e144.
Bendicho, S., Barbosa-Cánovas, G. V., & Martı́n, O. (2003a). Reduc-
tion of protease activity in milk by continuous flow high-intensity
pulsed electric field treatments. Journal of Dairy Science, 86,
697e703.
Bendicho, S., Barbosa-Cánovas, G. V., & Martı́n, O. (2003b). Reduc-
tion of protease activity in simulated milk ultrafiltrate by continu-
ous flow high intensity pulsed electric field treatments. Journal of
Food Science, 68(3), 952e957.
Bendicho, S., Espachs, A., Arántegui, J., & Martı́n, O. (2002). Effect of
high intensity pulsed electric fields and heat treatments on vitamins
of milk. Journal of Dairy Research, 69, 113e123.
Bendicho, S., Estela, C., Giner, J., Barbosa-Cánovas, G. V., &
Martı́n, O. (2002). Effects of high intensity pulsed electric field and
thermal treatments on a lipase from Pseudomonas fluorescens.
Journal of Dairy Science, 85, 19e27.
Castro, A. J., Swanson, B. G., Barbosa-Cánovas, G. V., & Zhang, Q. H.
(2001). Pulsed electric field modification of milk alkaline phos-
phatase activity. In G. V. Barbosa-Cánovas, & Q. H. Zhang (Eds.),
Pulsed electric fields in food processing (pp. 65e82). Lancaster:
Technomic Publishing.
Chen, W., & Wu, W. (2006). Electric field induced changes in
membrane proteins charge movement currents. Burns, 32(7),
833e841.
Cortés, C., Esteve, M. J., Rodrigo, D., Torregrosa, F., & Frı́gola, A.
(2006). Changes of colour and carotenoids contents during high
intensity pulsed electric field treatments in orange juices. Food and
Chemical Toxicology, 44, 1932e1939.
Cortés, C., Torregrosa, F., Esteve, M. J., & Frı́gola, A. (2006). Caroten-
oid profile modification during refrigerated storage in untreated
and pasteurized orange juice and orange juice treated with high-
intensity pulsed electric fields. Journal of Agricultural and Food
Chemistry, 54, 6247e6264.
 Tóth-Markus, M., & Lechner, N. (2006).
Cserhalmi, Zs, Sass-Kiss, A.,
Study of pulsed electric field treated citrus juices. Innovative Food
Science and Emerging Technologies, 7, 49e54.
Dörnenburg, H., & Knorr, D. (1993). Cellular permeabilisation of
cultured plant tissues by high electric field pulses or ultra high
pressure for the recovery of secondary metabolites. Food Biotech-
nology, 7(1), 35e48.
Elez-Martı́nez, P., & Martı́n-Belloso, O. (2007). Effects of high intensity
pulsed electric field processing conditions on vitamin C and anti-
oxidant capacity of orange juice and gazpacho, a cold vegetable
soup. Food Chemistry, 102, 201e209.
Elez-Martı́nez, P., Soliva-Fortuny, R. C., & Martı́n-Belloso, O. (2006).
Comparative study on shelf-life of orange juice processed by high
intensity pulsed electric fields or heat treatment. European Food
Research and Technology, 222(3e4), 321e329.
Elez-Martı́nez, P., Aguiló-Aguayo, I., & Martı́n-Belloso, O. (2006). In-
activation of orange juice peroxidase by high-intensity pulsed
electric fields as influenced by process parameters. Journal of the
Science of Food and Agriculture, 86, 71e81.
Elez-Martı́nez, P., Suárez-Recio, M., & Martı́n-Belloso, O. (2007).
Modeling the reduction of pectin methyl esterase activity in orange
juice by high intensity pulsed electric fields. Journal of Food En-
gineering, 78(1), 184e193.
Eshtiaghi, M. N., & Knorr, D. (2002). High electric field pulse pre-
treatment: potential for sugar beet processing. Journal of Food En-
gineering, 52, 265e272.
Espachs-Barroso, A., van Loey, A., Hendrickx, M., & Martı́n-
Belloso, O. (2006). Inactivation of plant pectin methylesterase by
thermal or high intensity pulsed electric field treatments.
Innovative Food Science and Emerging Technologies, 7(1e2),
40e48.
Evrendilek, G., Jin, Z. T., Ruhlman, K. T., & Qiu, X. (2000). Microbial
safety and shelf-life of apple juice and cider processed by bench
and pilot scale PEF systems. Innovative Food Science and Emerging
Technologies, 2000(1), 77e86.
Evrendilek, G. A., Yeom, H. W., Jin, Z. T., & Zhang, Q. H. (2004).
Safety and quality evaluation of yogurt-based drink processed by
a pilot plant PEF system. Journal of Food Process Engineering,
27(3), 197e212.
Fernandez-Diaz, M. D., Barsotti, L., Dumay, E., & Cheftel, J. C. (2000).
Effects of pulsed electric fields on ovalbumin solutions and dia-
lyzed egg white. Journal of Agricultural and Food Chemistry, 48(6),
2332e2339.
Ferruzzi, M. G., Böhm, V., Courtney, P. D., & Schwartz, S. J. (2002).
Antioxidant and antimutagenic activity of dietary chlorophyll de-
rivatives determined by radical scavenging and bacterial reverse
mutagenesis assays. Journal of Food Science, 67(7), 2589e2595.
Fiala, A., Wouters, P. C., van den Bosch, E., & Creyghton, Y. L. M.
(2001). Coupled electrical-fluid model of pulsed electric field
treatment in a model food system. Innovative Food Science and
Emerging Technologies, 2, 229e238.
Fincan, M., DeVito, F., & Dejmek, P. (2004). Pulsed electric field
treatment for solid-liquid extraction of red beetroot pigment.
Journal of Food Engineering, 64, 381e388.
Garde-Cerdán, T., Arias-Gil, M., Marsellés-Fontanet, A. R., Ancı́n-
Azpilicueta, C., & Martı́n-Belloso, O. (2007). Effects of thermal and
non-thermal processing treatments on fatty acids and free amino-
acids of grape juice. Food Control, 18, 473e479.
Gerlach, D., Alleborn, N., Baars, A., Delgado, A., Moritz, J., & Knorr,
D. (2008). Numerical simulations of pulsed electric fields for food
preservation: a review. Innovative Food Science & Emerging
Technologies, 9(4), 408e417.
Geulen, M., Teichgraeber, P., & Knorr, D. (1994). Zellaufscluss durch
elektrische hochspannungsimpulse. Lebensmitteltechnik, ZFL,
45(7/8), 24e27.
Giner, J., Gimeno, V., Barbosa-Cánovas, G., & Martı́n, O. (2001). Effects
of pulsed electric field processing on apple and pear polyphenolox-
idases. Food Science and Technology International, 7(4), 339e345.
Giner, J., Gimeno, V., Espachs, A., Elez, P., Barbosa-Cánovas, G. V., &
Martı́n, O. (2000). Inhibition of tomato (Licopersicon esculentum
Mill.) pectin methylesterase by pulsed electric fields. Innovative
Food Science and Emerging Technologies, 1(1), 57e67.
Giner, J., Ortega, M., Mesegué, M., Gimeno, V., Barbosa-
Cánovas, G. V., & Martı́n, O. (2002). Inactivation of peach poly-
phenoloxidase by exposure to pulsed electric fields. Journal of
Food Science, 67(4), 1467e1472.
Giner, J., Rauret-Ariño, A., Barbosa-Canovas, G. V., & Martin-Belloso,
O. (1997). Inactivation of polyphenoloxidase by pulsed electric
fields. In Proceedings IFT Annual meeting, p. 19, Orlando, USA.
Grahl, T., & Märkl, H. (1996). Killing of microorganisms by pulsed
electric fields. Applied Microbiology and Biotechnology, 45,
148e157.
Guderjan, M., Elez-Martinez, P., & Knorr, D. (2007). Application of
pulsed electric fields at oil yield and content of functional
food ingredients at the production of rapeseed oil. Innovative Food
Science and Emerging Technologies, 8, 55e62.
Guderjan, M., Toepfl, S., Angersbach, A., & Knorr, D. (2005). Impact of
pulsed electric field treatment on the recovery and quality of plant
oils. Journal of Food Engineering, 67(3), 281e287.
Heinz, V., Alvarez, I., Angersbach, A., & Knorr, D. (2002). Preservation
of liquid foods by high intensity pulsed electric fields-basic con-
cepts for process design. Trends in Food Science and Technology,
12, 103e111.
 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Ho, S. Y., Mittal, G. S., & Cross, J. D. (1997). Effects of high field
electric pulses on the activity of selected enzymes. Journal of Food
Engineering, 31, 69e84.
Hodgins, A. M., Mittal, G. S., & Griffiths, M. W. (2002). Pasteurization
of fresh orange juice using low energy pulsed electrical field.
Journal of Food Science, 67(6), 2294e2299.
Jeantet, R., Baron, F., Nau, F., Roignant, M., & Brulé, G. (1999). High
intensity pulsed electric fields applied to egg white: effect on Sal-
monella enteritidis inativation and protein denaturation. Journal of
Food Protection, 62(12), 1381e1386.
Jia, M., Zhang, H. Q., & Min, D. B. (1999). Pulsed electric field pro-
cessing effects on flavor compounds and microorganisms of orange
juice. Food Chemistry, 65(4), 445e451.
Jin, Z. T., & Zhang, Q. H. (1999). Pulsed electric field inactivation of
microorganisms and preservation of quality of cranberry juice.
Journal of Food Processing and Preservation, 23(6), 481e497.
Lindgren, M., Aronsson, K., Galt, S., & Ohlsson, T. (2002). Simulation
of the temperature increase in pulsed electric field (PEF) continuous
flow treatment chambers. Innovative Food Science & Emerging
Technologies, 3, 233e245.
Li, S.-Q., Bomser, J. A., & Zhang, Q. H. (2005). Effects of pulsed
electric fields and heat treatment on stability and secondary
structure of bovine immunoglobulin G. Journal of Agricultural and
Food Chemistry, 53, 663e670.
Li, Y., & Chen, Z. (2006). Effect of high intensity pulsed electric field on the
functional properties of protein isolated from soybean. Transactions
of the Chinese Society of Agricultural Engineering, 22(8), 194e198.
Ma, L., Chang, F. J., Góngora-Nieto, M. M., Barbosa-Cánovas, G. V., &
Swanson, B. G. (2001). Comparison study of pulsed electric fields,
high hydrostatic pressure, and termal processing on the electro-
phoretic patterns of liquid whole eggs. In G. V. Barbosa-Cánovas, &
Q. H. Zhang (Eds.), Pulsed electric fields in food processing
(pp. 225e240). Lancaster: Technomic Publishing.
Martı́n-Belloso, O., & Elez-Martı́nez, P. (2005). Enzymatic inactivation
by pulsed electric fields. In D-W. Sun (Ed.), Emerging technologies
for food processing (pp. 155e181). London: Elsevier Academic
Press.
Martı́n-Belloso, O., Vega-Mercado, H., Qin, B. L., Chang, F. J., Barbosa-
Cánovas, G. V., & Swanson, B. G. (1997). Inactivation of Escherichia
coli suspended in liquid egg using pulsed electric fields. Journal of
Food Processing and Preservation, 21(3), 193e208.
Min, S., & Zhang, Q. H. (2003). Effects of commercial-scale pulsed
electric field processing on flavor and color of tomato juice. Journal
of Food Science, 68(5), 1600e1606.
Min, S., Jin, Z. T., & Zhang (2003). Commercial scale pulsed electric
field processing of tomato juice. Journal of Agricultural and Food
Chemistry, 51(11), 3338e3344.
Min, S., Jin, Z. T., Min, S. K., Yeom, H., & Zhang, Q. H. (2003).
Commercial-scale pulsed electric field processing of orange juice.
Journal of Food Science, 68(4), 1265e1271.
Min, S., Min, S. K., & Zhang, Q. H. (2003). Inactivation kinetics of
tomato juice lipoxygenase by pulsed electric fields. Journal of Food
Science, 68(6), 1995e2001.
Morren, J., Roodenburg, B., & Han, S. W. H. D. (2003). Electro-
chemical reactions and electrode corrosion in pulsed electric field
(PEF) treatment chambers. Innovative Food Science & Emerging
Technologies, 4(3), 285e295.
Odriozola-Serrano, I., Bendicho-Porta, S., & Martı́n-Belloso, O.
(2006). Comparative study on shelf-life of whole milk processed by
high-intensity pulsed electric field or heat treatment. Journal of
Dairy Science, 89, 905e911.
Perez, O. E., & Pilosof, A. M. R. (2004). Pulsed electric fields effects on
the molecular structure and gelatin of b-lactoglobulin concentrate
and egg white. Food Research International, 37(1), 102e110.
Qiu, X., Sharma, S., Tuhela, L., Jia, M., & Zhang, Q. (1998). An inte-
grated PEIAC pilot plant for continuous nonthermal pasteurization
of fresh orange juice. Agricultural Engineering, 41, 1069e1074.
555
Rodrigo, D., Barbosa-Cánovas, G. V., Martı́nez, A., & Rodigro, M.
(2003). Pectin methyl esterase and natural micro flora of fresh
mixed orange and carrot juice treated with pulsed electric fields.
Journal of Food Protection, 66(12), 2336e2342.
Sánchez-Moreno, C., Plaza, L., Elez-Martı́nez, P., De Ancos, B.,
Martı́n-Belloso, O., & Cano, M. P. (2005). Impact of high pressure
and pulsed electric fields on bioactive compounds and antioxidant
activity of orange juice in comparison with traditional thermal
processing. Journal of Agricultural and Food Chemistry, 53,
4403e4409.
Schilling, S., Alber, T., Toepfl, S., Neidhart, S., Knorr, D., Schieber, A.,
et al. (2007). Effects of pulsed electric field treatment of apple mash
on juice yield and quality attributes of apple juices. Innovative
Food Science and Emerging Technologies, 8, 127e134.
Sharma, S. K., Zhang, Q. H., & Chism, G. W. (1998). Development of
a protein fortified fruit beverage and its quality when processed
with pulsed electric field treatment. Journal of Food Quality, 21(6),
459e473.
Taiwo, K. A., Angersbach, A., & Knorr, D. (2002). Influence of high
intensity electric field pulses and osmotic dehydration on the re-
hydration characteristics of apple slices at different temperatures.
Journal of Food Engineering, 52, 185e192.
Tedjo, W., Eshtiaghi, M. N., & Knorr, D. (2002). Einsatz, nicht-ther-
mischer verfahren zur zell-permeabilisierung von weintrauben ung
gewinnung von inhaltsstoffen. Fluessiges Obst, 9, 578e583.
Toepfl, S., Mathys, A., Heinz, V., & Knorr, D. (2006). Potential of high
hydrostatic pressure and pulsed electric fields for energy efficient
and environmentally friendly food processing. Food Reviews In-
ternational, 22, 405e423.
Torregrosa, F., Cortés, C., Esteve, M. J., & Frı́gola, A. (2005). Effect of
high-intensity pulsed electric fields processing and conventional
heat treatment on orange-carrot juice carotenoids. Journal of Ag-
ricultural and Food Chemistry, 53, 9519e9525.
Van Loey, A., Verachtert, B., & Hendrickx, M. (2002). Effects of high
electric field pulses on enzymes. Trends in Food Science and
Technology, 12, 94e102.
Vega-Mercado, H., Powers, J. R., Barbosa-Cánovas, G. V., &
Swanson, B. G. (1995). Plasmin inactivation with pulsed electric
fields. Journal of Food Science, 60, 1143e1146.
Vega-Mercado, H., Powers, J. R., Barbosa-Cánovas, G. V.,
Luedecke, L., & Swanson, B. G. (2001). Change in susceptibility of
proteins to proteolysis and the inactivation of an extracellular
protease from Pseudomonas fluorescens M3/6 when exposed to
pulsed electric fields. In G. V. Barbosa-Cánovas, & Q. H. Zhang
(Eds.), Pulsed electric fields in food processing (pp. 105e120).
Lancaster: Technomic Publishing.
Wu, Y., Mittal, G. S., & Griffiths, M. W. (2005). Effect of pulsed electric
field on the inactivation of microorganisms in grape juices with and
without antimicrobials. Biosystems Engineering, 90(1), 1e7.
Yang, R., Li, S.-Q., & Zhang, Q. H. (2004a). Effects of pulsed electric
fields on the activity of enzymes in aqueos solution. Journal of
Food Science, 69(4), 241e248.
Yang, R., Li, S.-Q., & Zhang, Q. H. (2004b). Effects of pulsed electric
fields on the activity and structure of pepsin. Journal of Agricultural
and Food Chemistry, 52(24), 7400e7406.
Ye, H., Huang, L. L., Chen, S. D., & Zhong, J. J. (2004). Pulsed electric
field stimulates plant secondary metabolism in suspension cultures
of Taxus chinensis. Biotechnology and Bioengineering, 88,
788e795.
Yeom, H. W., Streaker, C. B., Zhang, Q. H., & Min, D. B. (2000). Ef-
fects of pulsed electric fields on the quality of orange juice and
comparison with heat pasteurization. Journal of Agricultural and
Food Chemistry, 48(10), 4597e4605.
Yeom, H. W., Streaker, C. B., & Zhang, Q. H. (2000). Effects of pulsed
electric fields on the activities of microorganisms and pectin
methyl esterase in orange juice. Journal of Food Science, 65(8),
1359e1363.
556 R. Soliva-Fortuny et al. / Trends in Food Science & Technology 20 (2009) 544e556
Yeom, H. W., Zhang, Q. H., & Dunne, P. (1999). Inactivation of papain
by pulsed electric fields in a continuous system. Food Chemistry,
67, 53e59.
Yeom, H. W., & Zhang, Q. H. (2001). Enzymatic inactivation by pulsed
electric fields: a review. In G. V. Barbosa-Cánovas, & Q. H. Zhang
(Eds.), Pulsed electric fields in food processing (pp. 57e64).
Lancaster: Technomic Publishing.
Yeom, H. W., Zhang, Q. H., & Chism, G. W. (2002). Inactivation of
pectin methyl esterase in orange juice by pulsed electric fields.
Journal of Food Science, 67(6), 2154e2159.
Yin, Y., Han, Y., & Liu, J. (2007). A novel protecting method for
visual green color in spinach puree treated by high intensity
pulsed electric fields. Journal of Food Engineering, 79(4),
1256e1260.
Zhong, K., Wu, J., Wang, Z., Chen, F., Liao, X., Hu, X., et al. (2007).
Inactivation kinetics and secondary structural change of PEF-
treated POD and PPO. Food Chemistry, 100(1), 115e123.
Zimmermann, U., Pilwat, G., & Riemann, F. (1974). Dielectric
breakdown in cell membranes. Biophysical Journal, 14,
881e899.
Zulueta, A., Esteve, M. J., Frasquet, I., & Frı́gola, A. (2007). Fatty acid
profile changes during orange juice-milk beverage processing by
high-pulsed electric field. European Journal of Lipid Science and
Technology, 109(1), 25e31.