Pulsed Electric Field
J Raso, S Condón, and I Álvarez, Universidad de Zaragoza, Zaragoza, Spain
Ó 2014 Elsevier Ltd. All rights reserved.
Introduction
Pulsed electric fields (PEFs) is a technology that causes elec-
troporation of cell membranes by the application of intermit-
tent electric field strength of high intensity for periods of time
in the order of microseconds. Electroporation consists of the
increment of the cell membrane permeability to ions and
macromolecules. Such an increase in permeability is related to
the formation of local defects or pores in the cell membranes.
Depending on the intensity of the external electric field strength
applied, the viability of the electroporated cell can be preserved
by recovering the membrane integrity, or the electroporation
can be permanent, leading to cell death. Reversible electro-
poration is a procedure routinely used in molecular biology
and clinical biotechnological applications to gain access to the
cytoplasm in order to introduce or deliver in vivo drugs,
oligonucleotides, antibodies, plasmids, and the like. In the
1960s, it was demonstrated that irreversible electroporation
was an effective way to inactivate microorganisms, but it was
not until the end of the 1980s that there arose increased interest
in this technology as a nonthermal preservation method that
would reduce the undesirable changes induced by heat treat-
ments in foods. Efforts conducted in the last 20 years by
researchers in different fields (microbiology, chemistry, engi-
neering) have led to the development of PEF equipment on an
industrial scale and to the first commercial applications of PEF
technology for the preservation of premium quality fruit juices.
Technological Aspects of the Pulsed Electric Field
PEF is a treatment that involves the application of direct current
voltage pulses for very short periods of time, in the range
between microseconds to milliseconds, through a material
placed between two electrodes. This voltage results in an elec-
tric field whose intensity depends on the gap between the
electrodes and the voltage delivered.
Generation of Pulsed Electric Fields
Generation of PEFs requires a fast discharge of electrical energy
within a short period of time. The pulse generator and treat-
ment chamber are the basic components of an apparatus for
application of PEF (Figure 1). The pulse generator consists of
a charger that converts the AC to DC current and charges an
energy storage device such as a capacitor. The discharge of the
electrical energy in the treatment chamber is controlled by
a switch that is the key component in a pulsed system, because
it imposes a practical limitation of the pulse generator on
power and voltage level. High-power switches that have only
turn-on capability (e.g., spark gap) required the use of circuits
with a small energy storage tank. In these circuits the pulse that
is started by closing the switch will end when the storage tank is
empty. Food conductivity and circuit parameters, including
treatment chamber geometry, determine the rate at which the
966 Encyclopedia of Food Microbiology, Volume 2
depletion of the tank occurs, and, therefore, the duration of the
pulse. Switches with turn-on and turn-off capability are
increasingly used because they add flexibility to the pulse
generator. Here, the pulse duration is determined by the
control signal of the switch, because the pulse starts by closing
the switch and it ends by opening it.
The treatment chamber is composed of two electrodes held
in position by insulating material, which forms an enclosure
containing the food material. Electrodes should be designed to
minimize the effect of electrolysis as well as corrosion. Stainless
steel is commonly used as electrode material, but recently it has
been observed that titanium electrodes show a superior resis-
tance against corrosion. Static treatment chambers with parallel
electrodes are generally used for basic studies aimed to get
a fundamental understanding of the microbial inactivation by
PEF. However, the development of continuous flow treatment
chambers for PEF processing is essential for scaling up the
technology for nonthermal microbial inactivation. Although
several different designs have been developed in the last few
years, the two most important treatment chamber designs that
are being considered for commercial application of PEF are
parallel electrode and colinear configurations (Figure 2).
Parallel electrode configuration is the simplest chamber
geometry and consists of a rectangular duct of insulating
material with two electrodes on opposite sides. This configu-
ration imparts a uniform electric field in the treatment zone.
However, for some applications, a colinear configuration is
preferred because the load resistance of the treatment chamber
is higher and the energetic requirements are consequently
lower. The colinear treatment chamber consists of an electri-
cally insulating tube through which liquid flows. On either side
of this chamber, the electrodes are located. They consist of two
metal pipes that also serve as the entrance and exit for the fluid.
The circular section of colinear configuration facilitates its
installation in the circulation pipes used in the food industry.
However, the main problem of this configuration is the inho-
mogeneity in the electric field strength and temperature
distribution in the treatment chambers during PEF processing,
observed by numerical simulation techniques. Generation of
turbulent flow by modifying the treatment chamber geometry
Figure 1 Basic components of a PEF apparatus.
http://dx.doi.org/10.1016/B978-0-12-384730-0.00397-9

Figure 2 Treatment chamber configurations for continuous PEF processing and electric field strength distribution.
or by inserting a grid before the treatment zone has been sug-
gested to improve the treatment uniformity in colinear
configurations.
Definition of Process Parameters
The most typical process parameters that characterize PEF
technology are electric field strength, pulse shape, treatment
time pulse width, number of pulses, pulse-specific energy, and
frequency (Figure 3).
Electric field strength: The distance between the electrodes of
the treatment chamber and the voltage delivered defines the
strength of the electric field, which is generally reported in
kV cm
1. While in treatment chambers with parallel electrode
configurations, the electric field strength between the electrodes
is uniform, in colinear configuration the electric field strength is
not uniform and changes depending on the location.
Pulse shape: Depending on type of switch and the configu-
ration of the discharge circuit, several pulse shapes are possible,
but the main ones used are exponential decay and square wave
pulses. Exponential decay pulses are generated by high-power
switches that have only turn-on capability, and therefore,
discharge the total energy stored in the capacitor bank. These
pulses have a drastic surging rate, but a very slow decaying rate,
causing a long tail section that is ineffective in killing micro-
organisms and yields extra heat. Square waveform pulses can
be obtained by an incomplete discharge of a capacitor by
a switch with on/off capability or by using a more complex
pulse-forming network. These pulses are more suitable for PEF
microbial inactivation because they produce stable peak
voltage for the pulse duration. Both exponential and square
wave pulses can be unipolar or bipolar. It has been reported
that bipolar pulses may reduce unwanted electrolysis and the
deposition of food particles on the electrode surface.
NON-THERMAL PROCESSING j Pulsed Electric Field 967
Treatment time: Treatment time is defined as a function of
the duration of pulse width and the number of pulses applied.
It is generally reported in ms. In square waveform pulses, pulse
width corresponds to the duration of the pulse, but in expo-
nential decay pulses, the time required for the input voltage to
decay to 37% of its maximum value has been adopted as the
effective pulse width.
Specific energy of the pulse: This parameter depends on the
voltage applied, pulse width, and resistance of the treatment
chamber, which vary according to the geometry and conduc-
tivity of the material treated. It is commonly reported in
kJ kg
1. This parameter makes it possible to evaluate the energy
costs of the PEF process and, consequently, to compare the PEF
treatment efficiency with other technologies. As all the electrical
energy delivered for generation of PEFs in the treatment
chamber is dissipated as heat and the residence time of food
materials in the treatment chamber during a PEF treatment
is lower than 1 s, this parameter permits estimating the incre-
ment of the temperature of a food as a consequence of the
treatment.
Frequency: This parameter indicates the number of pulses
applied by unit of time, and it is reported in Hz (pulses per
second). For an industrial exploitation with high flow rates and
short residence times, high pulse repetition rates are up to
several hundred hertz.
Mechanisms of Microbial Inactivation by the Pulsed
Electric Field
It is generally accepted that electroporation of the microbial
cytoplasmatic membrane plays a major role in cell death
caused by PEF. Maintenance of the integrity and functionality
of the cytoplasmatic membrane is vital for microorganisms
968 NON-THERMAL PROCESSING j Pulsed Electric Field
Figure 3 Main process parameters of PEF technology.
because it protects cells from the surrounding environment by
acting as a semipermeable barrier. The application of an
external electric field of high intensity induces the formation of
pores, increasing the permeability of the membrane to ions
and macromolecules. Different techniques such as electron
microscopy examination, measurement of leakage of
intracellular material, measurement of osmotic response, or
measurement of the uptake of fluorescent dyes studies have
demonstrated that PEF causes electroporation of the cyto-
plasmatic membrane of microorganisms. Observation by
electron microscopy of PEF-treated bacteria and yeasts has
revealed morphological alterations, such as increasing surface
roughness, ruptures in the membranes, disruption of organ-
elles, and even leakage of cellular contents. However, these
modifications affect only a small number of the cells observed
under the microscope, and a correspondence between the
frequencies of appearance of morphological alterations and the
loss of viability determined by counting survivors in plate has
not been demonstrated. It suggests that other pathways
different from the morphological changes observed under the
electronic microscope are involved in microbial inactivation by
PEF. However, the direct involvement of cytoplasmatic
membrane permeabilization in cell inactivation by PEF has
been established using other techniques. Measurements of the
osmotic response of microbial cells after application of a PEF
treatment reveal that microorganisms partially lost the ability
to plasmolize in a hypertonic medium. On the other hand,
a correlation between the number of microorganisms inacti-
vated by PEF and the measurement of the increased uptake of
fluorescent dyes such as propidium iodide unable to go
through intact membrane has been found, or intracellular UV-
absorbing material (nucleic acids, proteins) outside the cells
after PEF treatments has been detected.
The scheme represented in Figure 4 aims to summarize the
possible consequences derived from the electroporation of
a microbial cell by PEF. Early studies indicated that microbial
inactivation by PEF was an all-or-nothing effect because after
the treatment alive or dead cells were detected, but not
sublethal, injured ones. Sublethally injured cells are those
microorganisms that have suffered some kind of damage as
a result of the treatment applied, and they are able to recover
and grow only when the recovery conditions (temperature,
growth medium, etc.) are optimal. Presently, it is well estab-
lished that similar to other inactivation techniques, PEF causes
sublethal injury. The fact that the presence of sodium chloride
in the recovery medium prevents the growth of sublethally
injured cells after PEF treatment and the demonstration that
these damaged cells required the synthesis of lipids for repair
supports the involvement of the cytoplasmatic membrane in
cell injury. However, these observations cannot overlook the

Figure 4 Possible consequences derived from the electroporation of a microbial cell by PEF.
fact that sublethal injury was also caused by additional
damages of PEF in other microbial structures. The occurrence
of reversible permeabilization of microbial cytoplasmatic
membrane has been shown by comparing the uptake of pro-
pidium iodine when the dye is present in the treatment
medium during the PEF treatment or when it is added just
after the treatment. Therefore, although a percentage of the
microbial population may recover membrane integrity after
completing the PEF treatment, during the treatment the
increment of the cell membrane permeability may cause the
intake or uptake of ions and macromolecules in the microbial
cytoplasm, rendering damaged cells with an intact cyto-
plasmatic membrane.
Although a relationship between the increment of the
permeability of the cytoplasmatic membrane by application of
external electric field pulses and the microbial inactivation has
been demonstrated, very little is known about what is really
occurring in the membranes at the molecular level. Several
theories have been proposed to explain the mechanisms of
membrane electroporation. Electromechanical theories assume
that the external electric field applied causes membrane
compression, leading to membrane rupture when the electrical
force exceeds the elastic restoring force. From an electrical point
view, due to the low electrical conductivity of a membrane as
compared with the surrounding liquid, a cell can be considered
to resemble a spherical capacitor. When the cell is exposed to
external electric field strength, a time- and position-dependent
transmembrane potential would be induced across the cyto-
plasmic membrane because of the accumulation of oppositely
charged ions at both sides of the nonconductive membrane.
The attraction between these ions would cause membrane
thickness reduction and formation of pores. A critical value of
the external electric field is required to induce a transmembrane
potential (0.2–1.0 V) that leads to the formation of reversible
or irreversible pores in the membrane. When the applied
external electric field is around the critical value, reversible
electroporation would occur, allowing the cell membrane to
recover its structure and functionality. Irreversible electro-
poration resulting in membrane disintegration and loss of cell
viability are expected to occur when electric field strengths
higher than the critical value are applied.
NON-THERMAL PROCESSING j Pulsed Electric Field 969
Other theories assume that electroporation in a cell
membrane occurs both in protein channels and in the lipid
domain. According to these theories, an external electric field
may cause reorientation of lipid molecules of the membrane,
creating hydrophilic pores that could conduct current. Local
Joule heating generated by passage of electrical current would
induce thermal phase transition of the lipid bilayer. The
molecular dynamics of these events would involve changes in
conformation of lipid molecules and rearrangement of the
lipid bilayer by expanding the existing pores, creating new
hydrophobic pores and forming structurally more stable
hydrophilic pores. On the other hand, as the opening/closing
of many protein channels is dependent on transmembrane
potentials, it would be expected that when a PEF is applied,
many voltage-sensitive channel proteins would be opened.
Once these channels are opened, they would conduct higher
current than that for which they are designed; as result, these
channels would become irreversibly denatured by Joule heat-
ing or electrical modification of their functional groups.
The proposed theories to explain electropermeabilization
are based on experiments on model systems such as liposomes
or on individual eukaryote cells. However, in microorganisms,
the cytoplasmatic membrane is not the only envelope that
separates the cytoplasm from the environment. Yeast, bacteria,
and bacterial spores have additional structures such as the wall
cell (yeast and Gram-positive bacteria) and wall cell and
external membrane (Gram-negative bacteria) whose influence
in the membrane electroporation by PEF remains unknown. In
the case of bacterial spores, PEF fails to inactivate them,
probably because of the spores’ envelopes such as the coat and
the cortex, preventing the permeabilization effects of PEF on
the spore cytoplasmatic membrane.
Factors Affecting Microbial Inactivation by Pulsed
Electric Fields
The microbial resistance to a given processing technology that
acts by inactivating microorganisms has been found to depend
on many factors. In order to establish the process conditions to
ensure microbiological safety and stability, the influence of
970 NON-THERMAL PROCESSING j Pulsed Electric Field

Table 1 Relative significance of factors affecting microbial resistance to PEF

Process parameters Microbial characteristics Product parameters

Electric field strength *** Strain *** Composition ***

Treatment time *** Specie *** Conductivity *
Pulsed width * Growth conditions pH ***
Specific energy *** Growth temperature NMR aw NMR
Frequency * Growth phase **
Temperature *** Recovery conditions
Pulse shape * Medium composition ***
Temperature ***
Recovery time **
Oxygen concentration NMR

NMR: Need More Research.

***
Very significant.
**
Significant.
*Slightly significant.

these factors on microbial inactivation must be understood. 0

Log10 survival fraction (N/N0)

Factors affecting microbial inactivation by PEF have been
classified into three groups: processing parameters, microbial –1
characteristics, and treatment medium characteristics (Table 1). –2
The relative influence of these factors on microbial resistance to
–3
PEF is shown in Table 1.
–4

Processing Parameters –5

Among the process parameters, electric field strength and –6

treatment time are critical to the effectiveness of microbial
inactivation by PEF. Microbial inactivation increases by
increasing the strength of the electric field over a threshold
field strength called critical electric field strength (Ec). The Ec
differs for different microorganisms, but overall to obtain
significant microbial destruction, electric field strengths above
5 kV cm
1 are generally required. Generally, studies on
microbial inactivation have been conducted from 10 to
30 kV cm
1 because applications of higher electric field
strengths have technical limitations, especially at the indus-
trial scale, and may cause the dielectric breakdown of the food
material.
In general, PEF lethality increases with the treatment time.
The survival curves at constant electric field strength are char-
acterized by a fast inactivation in the first moments of the
treatment, and then the number of survivors slowly
decreases as the number of pulses applied becomes longer
(Figure 5).
It has been observed that PEF microbial inactivation
increases with the high-voltage electrical energy applied per
mass unit (specific energy). The specific energy has been
proposed as a control parameter of the PEF process, especially
when exponential decay pulses are used because of the lack of
precision in the measurement of the pulse width. It has been
reported that when applying different treatments of the same
specific energy by changing the electric field strength and
treatment time, those applied at higher electric fields are more
effective in terms of microbial inactivation. Therefore, to
characterize a PEF treatment, both specific energy and specific
energy should be reported together.
Some controversy has arisen concerning the influence
of the pulse shape, width, and frequency on PEF microbial
–7
0 25 50 75 100 125 150
Time ( s)
Figure 5 Theoretical survival curves corresponding to microbial inac-
tivation by PEF treatments at different electric field strengths (15 kV cm
1
(n), 20 kV cm
1 (:), 25 kV cm
1 (;), 30 kV cm
1 (A), and
35 kV cm
1 (l)).
inactivation. It is generally accepted that square wave pulses are
better than exponential decay ones because the slow decaying
rate causes a long tail section that is ineffective to kill the
microorganisms in the food material and yields extra heat.
Some authors have reported that when treatments of the same
duration are applied with pulses of different widths or at
different frequencies, longer pulses and higher frequencies are
more effective. However, these two parameters apparently exert
no influence on microbial inactivation when the temperature
rise of the medium caused by the application of longer pulses
of higher frequencies is avoided.
Microbial inactivation by PEF is usually enhanced when the
temperature of the treatment medium is increased, even in
ranges of temperatures that are not lethal for microorganisms
(Figure 6). This effect has been attributed to changes in the
phospholipid bilayer structure of the cell membranes, from
a gel-like consistency to a liquid crystalline state that is caused
by the temperature increase. Recently, it has been demonstrated
that the application of PEF treatments at moderate tempera-
tures (>50  C) introduces the possibility of pasteurizing liquid
foods by using short treatments at moderate electric field
strengths (25 kV cm
1).
 NON-THERMAL PROCESSING j Pulsed Electric Field 971

7 5 pH 4.0
pH 7.0
6

Log10 cycles of inactivation

Log10 cycles of inactivation
4
5
3
4

3 2

2
1

1
0
0

32
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2
1
7
10

67

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36

03
47
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7:
ºC

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ºC

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.5

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.5

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c.

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15

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27
15

38

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4

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Figure 6 Influence of the temperature of the treatment medium on the
lethality of Salmonella typhimurium by a PEF treatment (30 kV cm
1, 0.5 Hz,
square wave pulses of 3 ms) in media of pH 3.5. Error bars correspond
to standard deviation. Adapted from Saldaña, G., Puértolas, E., Álvarez, I.,
Meneses, N., Knorr, D., Raso, J., 2010. Evaluation of a static treatment
chamber to investigate kinetics of microbial inactivation by pulsed electric
fields at different temperatures at quasi-isothermal conditions. Journal of
Food Engineering 100, 349–356.
Microbial Characteristics
Microbial inactivation by PEF depends on microbial properties
such as the type of microorganism, characteristics of the cell
envelopes (Gram-positive or negative), cell size, and shape.
Generally, it has been reported that bacteria are more PEF
resistant than yeast, Gram-negative microorganisms are more
sensitive than Gram-positive microorganisms, and coccus
bacteria are more resistant than rods. However, it seems that
the intrinsic microbial resistance is more important than the
effect of the microbial characteristics in determining the
microbial sensitivity to PEF. When the PEF resistance of
different microorganisms is compared under the same experi-
mental conditions, it is observed that some yeast cells are more
PEF resistant than some bacteria, some Gram-positive micro-
organisms are more sensitive than some Gram-negative
microorganisms, and some yeast species and some rod bacteria
are more resistant than some coccus bacteria.
Several studies have demonstrated that the PEF resistance of
different strains of bacterial species may vary greatly. It has been
observed that depending on the strain and pH of the treatment
medium, the inactivation of different strains of the same
microorganism may range from 0.1 to 4.5 log10 CFU m
1
(Figure 7). As the PEF resistance of the different strains
depended on the pH of the treatment medium, the target
microorganisms to define treatment conditions for PEF
pasteurization could be expected to be different for foods,
depending on their pH.
Treatment Medium Characteristics
Generally, studies on microbial inactivation by PEF have
been conducted with microorganisms suspended in liquid
media. The effect of PEF treatments on the inactivation of
c.
E.
Figure 7 Variability in the PEF resistance (30 kV cm
1; 50 pulses of
3 ms) of different strains of Escherichia coli (E.c.) and Listeria mono-
cytogenes (L.m.) treated in media of pH 4.0 (black bars) and 7.0 (white
bars). Adapted from Saldaña, G., Puértolas, E., López, N., García, D.,
Álvarez, I., Raso, J., 2009. Comparing the PEF resistance and occurrence
of sublethal injury on different strains of Escherichia coli, Salmonella
typhimurium, Listeria monocytogenes and Staphylococcus aureus in media
of pH 4 and 7. Innovative Food Science and Emerging Technologies
10, 160–165.
microorganisms in solid media or in media containing parti-
cles has received less attention. The influence of the electrical
conductivity and the pH of the substrate on microbial inacti-
vation has been widely investigated. Several studies concluded
that the conductivity of the treatment medium affects micro-
bial inactivation. However, these investigations do not make it
clear if the conductivity influences the effect of the electric field
on microbial membrane, or if the effect observed is a conse-
quence of the influence of conductivity on the characteristics of
the PEF treatment applied. A change in conductivity modifies
the resistance of the treatment chamber, and as a result it may
cause changes in the electric field strength and the pulse width
and total specific energy of the pulses. In a range of conductivity
from 0.5 to 4.0 mS cm
1, which corresponds to the conduc-
tivity of most liquid foods, it has been observed that the
conductivity did not affect microbial inactivation when the
input voltage and input pulse width were modified in order to
obtain the same treatment (field strength and treatment time)
in media of different conductivities.
Published research indicates that microbial PEF resistance
varies considerably depending on the pH of the treatment
medium. Researchers have reported that a variation of the pH
of the treatment medium can increase, reduce, or have no effect
on modifying the microbial sensitivity to PEF. Generally,
Gram-positive microorganisms are more PEF resistant in media
of neutral pH than in acidic conditions, and Gram-negative
ones are more resistant in media of acidic pH than in neutral
conditions. This effect of the pH on microbial resistance has
been confirmed in both buffers and liquid foods. The mecha-
nism that explains these differences seems to be related to the
occurrence of sublethal membrane damage by PEF. It has been
observed that when Gram-positive bacteria are treated in
972 NON-THERMAL PROCESSING j Pulsed Electric Field
neutral media, their ability to repair sublethal injury caused by
PEF is higher than when treated in low pH media. On the
contrary, the higher PEF ability to repair sublethal injury in
Gram-negative bacteria occurs when they are treated in acidic
pH media.
The influence of the aw of the treatment medium on
microbial inactivation has scarcely been investigated, and the
effect of the type of solute used to reduce the aw is unclear. Few
studies conducted indicate that a decrease in the aw increased
microbial PEF resistance. This effect has been explained by
a reduction of the cell volume and/or changes in the thickness,
permeability, and fluidity of the microbial membrane when
microorganisms are transferred to an environment with
lower aw.
The possible protection or sensitization to electric fields
conferred by different food components, such as carbohydrates,
lipids, or proteins, has been investigated for different authors.
However, the different treatment conditions and media used
make it difficult to obtain definitive conclusions in this respect.
For example, while some authors determined that microbial
resistance increased with the fat content of milk, others found
that microbial inactivation was independent of the fat or protein
content when buffers were used as treatment media. On the
other hand, a protective effect that made Escherichia coli more
PEF resistant has been reported as a consequence of the presence
of organic acids in both buffers of pH 4 and fruit juices. Further
research is necessary to determine the mechanism involved in
the protective effects observed.
Kinetics of Microbial Inactivation by Pulsed Electric
Field
A description of the kinetics of microbial inactivation by PEF
and quantification of how different factors can influence the
speed of microbial death is required for the development of
predictive models.
Some of the first investigations on microbial inactivation by
PEF suggested a linear relationship between the log of survivors
and the treatment times describe the survival curves. However,
later on it was observed that when the treatment time was
prolonged to achieve higher inactivation, the shape of the
survival curves was generally concave upward. Several equa-
tions have been proposed to describe these survival curves, but
presently an equation based on the Weibull distribution is the
most frequently used, because of its simplicity and flexibility.
Generally, the secondary models that are used to describe the
microbial inactivation by PEF are based on quadratic equations
whose complexity increases with the number of processing
variables investigated and the experimental range considered.
Recently, the combination of experimental design techniques
with multiple regression analysis and Monte Carlo simulation
have been used to establish the most influential factors on the
inactivation of pathogenic microorganisms by PEF.
Predictive models are useful tools for product development
and PEF process design in order to define the treatment
condition required for the food materials to meet specifications
for safety and stability; establish the requirements that the PEF
equipment must meet to apply the treatment on a commercial
scale; or conduct a cost analysis of the processing options so that
120 0
Input energy (kJ kg–1)
100
Treatment time ( s)
40
35 ºC
80 30 ºC
80
25 ºC
60
20 ºC
120
20 ºC
40 25 ºC
30 ºC
160
20 35 ºC
0 200
20 22 24 26 28 30
Electric field strength (kV cm–1)
Figure 8 Example of application of mathematical modeling for optimi-
zation treatment conditions. Treatment time (black lines) and specific
energy input (red lines) required to inactivate 5 log10 cycles the population
of E. coli O157:H7 suspended in apple juice by PEF at different electric field
strengths and initial treatment temperatures. Dotted lines indicate an
example of the input energy and treatment time required for 5 log10
reductions at 24 kV cm
1 and an inlet temperature of 25  C. Adapted from
Saldaña, G., Puértolas, E., Monfort, S., Raso, J., and Álvarez, I., 2011.
Defining treatment conditions for PEF pasteurization of apple juice.
International Journal of Food Microbiology 151, 29–35.
production can run as economically as possible while delivering
a microbiologically safe product to the consumer (Figure 8).
Microbial Inactivation by Combined Processes
Including Pulsed Electric Fields
Since microbial resistance by PEF is affected by many different
factors, in some cases very intense treatments are necessary to
obtain the microbial inactivation levels required for assuring
food safety and stability. Combining PEF with other preserva-
tion methods has been widely investigated in order to increase
the lethal effect of PEF. PEF, in combination with other phys-
ical methods of microbial inactivation based on thermal and
nonthermal effects (i.e., high hydrostatic pressure, high-
pressure carbon dioxide, ultrasound, ultraviolet radiation, or
high-intensity light pulses), has proven to be effective in
enhancing microbial inactivation. Generally, the combination
of these treatments has consisted of a successive application of
hurdles, and it has proven to cause microbial reductions higher
than 5 log10 cycles in different pathogenic microorganisms.
The combination of PEF with antimicrobials such as bacte-
riocins (e.g., nisin, enterocin AS-48), enzymes (e.g., lysozyme),
organic acids (e.g., citric, lactic, acetic, malic), or essential oils
(e.g., clove, carvacrol, citral) is another approach that is
considered to improve microbial lethality. Several authors
reported additive or synergistic effects on microbial inactivation
when antimicrobials were added to the treatment medium,
including buffers and liquid foods. However, it is necessary to
bear in mind that some antimicrobials used in these combi-
nations, such as nisin or lysozyme, are ineffective or scarcely
effective against Gram-negative bacteria: The impermeability of
the outer membrane of these bacteria does not allow these
antimicrobials to reach its site of action; the site of action for
lysozyme is the cell wall, and for nisin the cytoplasmatic
membrane.

Industrial Application Based on Microbial
Inactivation by Pulsed Electric Field
PEF’s ability to inactivate the vegetative cells of microorganisms
at temperatures that avoid the harmful effects of heat on the
organoleptic properties and the nutrient values of liquid foods
makes this technology very attractive for the food industry. As
bacterial spores are resistant to PEF treatments, applications of
PEF should be focused on pasteurization. Although the main
objective of PEF pasteurization is to guarantee food safety,
a large proportion of the population of vegetative spoilage
microorganisms is also inactivated by the treatment, contrib-
uting to extending the shelf life of foods. However, PEF is not
capable of achieving commercial sterility because spores or
other nonpublic health-significant microorganisms can be
present. Thus, other preservation techniques, such as refriger-
ation, atmosphere modification, the addition of preservatives,
or a combination of these techniques, will be required to
preserve the quality and stability of the food during its distri-
bution and storage.
The lack of reliable and economically viable industrial-scale
equipment limited the commercial exploitation of PEF in the
food industry. However, recent developments in pulse power
generators have permitted the design of compact, reliable PEF
equipment for liquid food pasteurization at flow rates from
1000 to 2000 l h
1. The first commercial PEF-processed prod-
ucts have been fruit juices and smoothies. The major benefits of
PEF processing include the extension of shelf life from 7 to
21 days while maintaining superior taste and freshness as
compared to thermal processing.
Although the first commercial applications of PEF are
available in the market, more multidisciplinary research efforts
are required to improve the distribution of the electric field
strength distribution in continuous flow treatment chambers, to
develop suitable sensors to assess the PEF process, to identify
the most PEF-resistant pathogens of concern for each specific
food, to define process criteria for PEF pasteurization, and to get
a better mechanistic understanding of the critical parameters
affecting microbial inactivation. A deeper knowledge of these
aspects is needed to satisfy regulatory agencies and to enhance
the safety and stability of minimal process foods of the future.
See also: Heat Treatment of Foods – Principles of Pasteurization;
Heat Treatment of Foods: Action of Microwaves; Minimal
Methods of Processing: Manothermosonication; Predictive
Microbiology and Food Safety; Ultraviolet Light;
Nonthermal Processing: Pulsed UV Light; Nonthermal
Processing: Irradiation; Nonthermal Processing: Microwave;
Nonthermal Processing: Ultrasonication; Nonthermal
Processing: Cold Plasma for Bioefficient Food Processing;
Thermal Processes: Pasteurization; Injured and Stressed
Cells; Fruit and Vegetable Juices.
NON-THERMAL PROCESSING j Pulsed Electric Field 973
Further Reading
Barbosa-Cánovas, G.V., Tapia, M.S., Cano, M.P., 2005. Novel Food Processing
Technologies. Marcel Dekker/CRC Press, Boca Raton, FL.
García, D., Gómez, N., Mañas, P., Condón, S., Raso, J., Pagán, R., 2005. Occurrence
of sublethal injury after pulsed electric fields depending on the micro-organism, the
treatment medium pH and the intensity of the treatment investigated. Journal of
Applied Microbiology 99, 94–104.
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 and Emerging Technologies 9, 408–417.
Lelieveld, H.L.M., Notermans, S., de Haan, S.W.H., 2007. Food Preservation by Pulsed
Electric Fields: From Research to Application. Woodhead, Abington.
Martín-Belloso, O., Sobrino-López, A., 2011. Combination of pulsed electric fields with
other preservation techniques. Food and Bioprocess Technology 4, 954–968.
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, 1265–1271.
Raso, J., Barbosa-Cánovas, G.V., 2003. Non-thermal preservation of foods using
combined processing techniques. Critical Reviews in Food Science and Nutrition
43, 265–285.
Raso, J., Heinz, V., 2006. Pulsed Electric Fields Technology for the Food Industry:
Fundamentals and Applications. Springer, New York.
Saldaña, G., Puértolas, E., López, N., García, D., Álvarez, I., Raso, J., 2009.
Comparing the PEF resistance and occurrence of sublethal injury on different
strains of Escherichia coli, Salmonella typhimurium, Listeria monocytogenes and
Staphylococcus aureus in media of pH 4 and 7. Innovative Food Science and
Emerging Technologies 10, 160–165.
Saldaña, G., Puértolas, E., Álvarez, I., Meneses, N., Knorr, D., Raso, J., 2010.
Evaluation of a static treatment chamber to investigate kinetics of microbial
inactivation by pulsed electric fields at different temperatures at quasi-isothermal
conditions. Journal of Food Engineering 100, 349–356.
Saldaña, G., Puértolas, E., Monfort, S., Raso, J., Álvarez, I., 2011. Defining treatment
conditions for PEF pasteurization of apple juice. International Journal of Food
Microbiology 151, 29–35.
Sampedro, F., Rodrigo, D., Martínez, A., 2011. Modelling the effect of pH and pectin
concentration on the PEF inactivation of Salmonella enterica serovar Typhimurium
by using the Monte Carlo simulation. Food Control 22, 420–425.
Saulis, G., 2010. Electroporation of cell membranes: the fundamental effects of pulsed
electric fields in food processing. Food Engineering Reviews 2, 52–73.
Toepfl, S., Heinz, V., Knorr, D., 2007. High intensity pulsed electric fields applied for
food preservation. Chemical Engineering and Processing 46, 537–546.
Wouters, P.C., Álvarez, I., Raso, J., 2001. Critical factors determining inactivation
kinetics by pulsed electric field food processing. Trends in Food Science and
Technology 12, 112–121.
Relevant Websites
Manufactures of PEF industrial scale equipment:
http://www.elcrack.de/.
http://purepulse.eu/.
http://www.divtecs.com/food-and-wastewater-processing/.