Journal Pre-proofs

Review

Pulsed electric field technology in vegetable and fruit juice processing: A re-
view

Iuri Procopio Castro Brito, Eric Keven Silva

PII: S0963-9969(24)00277-1
DOI: https://doi.org/10.1016/j.foodres.2024.114207
Reference: FRIN 114207

To appear in: Food Research International

Received Date: 20 December 2023

Revised Date: 22 February 2024
Accepted Date: 10 March 2024

Please cite this article as: Brito, I.P.C., Silva, E.K., Pulsed electric field technology in vegetable and fruit juice
processing: A review, Food Research International (2024), doi: https://doi.org/10.1016/j.foodres.2024.114207

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Pulsed electric field technology in vegetable and fruit juice processing: A review

Iuri Procopio Castro Brito and Eric Keven Silva*

Faculdade de Engenharia de Alimentos (FEA), Universidade Estadual de Campinas

(UNICAMP), Rua Monteiro Lobato, 80, Campinas-SP CEP:13083-862, Brazil.

*
Corresponding author: ekeven@unicamp.br.

1
Abstract: The worldwide market for vegetable and fruit juices stands as a thriving sector
with projected revenues reaching to $81.4 billion by 2024 and an anticipated annual growth
rate of 5.27% until 2028. Juices offer a convenient means of consuming bioactive compounds
and essential nutrients crucial for human health. However, conventional thermal treatments
employed in the juice and beverage industry to inactivate spoilage and pathogenic
microorganisms, as well as endogenous enzymes, can lead to the degradation of bioactive
compounds and vitamins. In response, non-thermal technologies have emerged as promising
alternatives to traditional heat processing, with pulsed electric field (PEF) technology
standing out as an innovative and sustainable choice. In this context, this comprehensive
review investigated the impact of PEF on the microbiological, physicochemical, functional,
nutritional, and sensory qualities of vegetable and fruit juices. PEF induces electroporation
phenomena in cell membranes, resulting in reversible or irreversible changes. Consequently,
a detailed examination of the effects of PEF process variables on juice properties is essential.
Monitoring factors such as electric field strength, frequency, pulse width, total treatment time,
and specific energy is important to ensure the production of a safe and chemically/kinetically
stable product. PEF technology proves effective in microbial and enzymatic inactivation
within vegetable and fruit juices, mitigating factors contributing to deterioration while
maintaining the physicochemical characteristics of these products. Furthermore, PEF
treatment does not compromise the content of substances with functional, nutritional, and
sensory properties, such as phenolic compounds and vitamins. When compared to alternative
processing methods, such as mild thermal treatments and other non-thermal technologies,
PEF treatment consistently demonstrates comparable outcomes in terms of physicochemical
attributes, functional properties, nutritional quality, and overall safety.

Keywords: Electroporation; microbial inactivation; phenolic compounds, ascorbic acid.

2
1. Introduction

The modern consumer has access to a wide range of information through various
communication channels, directly influencing their dietary choices. Therefore, new
market trends focused on health and sustainability have consolidated over the past decade.
Within this context, food is no longer solely seen as a source of nutrients. From the current
perspective, the food also must contribute to the proper functioning of the organism,
promoting health and well-being while presenting a high standard of sensory quality.
Consequently, processed food products should promote a sensory experience similar to
the fresh product (Fabjanowicz et al., 2024; Gupta et al., 2023; Lepaus et al., 2023;
Martins, Rosenthal, Ares, & Deliza, 2020; Nonglait, Chukkan, Arya, Bhat, & Waghmare,
2022; Salazar-Zúñiga et al., 2023). As a result of this, food industries must align their
production lines or operational methods with the demands of the modern consumer,
directly implying the adoption of new technologies and suitable processing approaches.

Within this context, non-thermal technologies have been investigated as a

promising alternative for food and beverage processing, as they meet the main demands
of the current consumer market (Martins et al., 2020; Nonglait et al., 2022; Salazar-
Zúñiga et al., 2023). These innovative technologies incorporate principles that enhance
safety and physicochemical properties without compromising the nutritional, functional,
and sensory aspects of the food products. Additionally, the role of these technologies in
environmental preservation stands out. The incorporation of non-thermal technologies in
food and beverage production represents a contribution to sustainability in the production
sector. Conventional thermal treatments contribute to global warming primarily due to
the release of greenhouse gas, particularly CO2. When fossil fuels, such as coal, oil and
natural gas, are burned for thermal energy in industrial processes or power generation,
they release large amounts of CO2 into the atmosphere. This process significantly adds to
the concentration of greenhouse gas, which trap heat and contribute to a greenhouse
effect.

On the other hand, non-thermal technologies enable the use of sustainable energy
sources such as wind, solar, biogas, among others. Pulsed electric field (PEF), high
intensity ultrasound (HIUS), high hydrostatic pressure (HPP) processing, supercritical
carbon dioxide (scCO2), among others, are examples of non-thermal technologies being
developed and consolidated in the food industry (Arya, Nachiappan, Waghmare, & Bhat,
2023; Ghoshal, 2023; M. A. Lima & Rosenthal, 2022; Rathnakumar et al., 2023; Silva,
Meireles, & Saldaña, 2020; Vignali, Gozzi, Pelacci, & Stefanini, 2022).

The global market for vegetable and fruit juices is a flourishing industry, expected
to achieve revenues of $81.4 billion by 2024, with a projected annual growth rate of
5.27% until 2028 (Statista, 2024). Vegetable and fruit juices are commonly consumed as
a means to incorporate health-promoting bioactive compounds into dietary habits of
people across various cultures worldwide (Gupta et al., 2023; M. A. Lima & Rosenthal,
2022; Ravichandran, Jayachandran, Kothakota, Pandiselvam, & Balasubramaniam,
2023). Moreover, juice consumption is convenient and facilitates the intake of nutrients,
vitamins, antioxidants, fibers, and phytochemical compounds found in vegetable and fruit
(Fabjanowicz et al., 2024; Liao et al., 2023). Thermal processes can induce various
physicochemical, nutritional, and sensory changes in these beverages, resulting in
significant differences compared to fresh juices (Arya et al., 2023; Nonglait et al., 2022;
Pandiselvam et al., 2023; Petruzzi et al., 2017). These differences between fresh and
3
thermally processed juice result from the degradation of substances that contribute to the
nutritional and functional value and sensory aspects of the products. Therefore,
considering the new demands of the consumer market, the design of non-thermal
stabilization processes capable of preserving the characteristics of fresh juice is a
promising strategy to produce beverages of a high technological standard.

Among the emerging non-thermal technologies, PEF is an innovative technology

that involves the application of electrical pulses in food processing. In this approach,
electric current is applied at short intervals, promoting changes in the microstructure of
the food by redistributing charges in cellular structures and macromolecules (Cavalcanti
et al., 2023; Ghoshal, 2023; Rostamabadi et al., 2023). These changes result in microbial
and enzymatic inactivation, providing microbiological safety and kinetic stability to the
beverage. Furthermore, PEF technology has various applications, such as for the recovery
of bioactive compounds in food matrices and agro-food waste, molecular complexation,
biopolymer modification, and serving as an assisting technology in freezing/thawing and
drying processes, and others (Achayuthakan, Wongsagonsup, Sriprablom,
Suphantharika, & Intra, 2023; Bernabeu, Salgado-Ramos, Barba, Collado, & Castagnini,
2024; Llavata, Collazos-Escobar, García-Pérez, & Cárcel, 2024; Nemli et al., 2024;
Nyoto & Gómez Galindo, 2023; Öztürk, Buzrul, Bilge, & Yurdakul, 2024). The electric
field can still preserve nutritional and functional products without compromising sensory
characteristics. Recent studies in the literature indicate PEF as a promising technology
for the non-thermal processing of fruit and vegetable juices (Akdemir Evrendilek, Keskin,
& Golge, 2020; Buitimea-Cantúa et al., 2022; Delso, Berzosa, Sanz, Álvarez, & Raso,
2022; Jin & Aboelhaggag, 2022; Mendes-Oliveira, Jin, & Campanella, 2022; Wibowo et
al., 2019; Yildiz, Pokhrel, Unluturk, & Barbosa-Cánovas, 2021).

Several studies on the impact of PEF processing on the safety, nutritional,

functional and sensory parameters of vegetable and fruit juices can be found in the
literature (Akdemir Evrendilek, Agcam, & Akyildiz, 2021; Delso, Berzosa, et al., 2022;
Dziadek et al., 2019; Lee et al., 2021; Rahaman et al., 2020; Wibowo et al., 2019; Yildiz
et al., 2021). Innovative processing technologies such as PEF are being assessed due to
the high demand for process that primarily preserve the health-beneficial and sensory
properties of juices. In this context, this review discussed the impacts of PEF technology
on vegetable and fruit juices processing. Challenges and opportunities of implementing
this non-thermal process in juice production were examined, reviewing the impacts of the
electric field on physicochemical, microbiological, nutritional, functional, and sensory
characteristics. Additionally, aspects related to the severity level of PEF treatments were
discussed.

2. Fundamentals of PEF technology

PEF is a non-thermal technology that causes the formation of pores in the cell
membrane. This phenomenon is called electroporation. The application of an electric field
generates charges in the membrane components. Charges of opposite polarity attract each
other, and charges of the same polarity repel each other. This creates spaces in the cell
membrane called pores (Naliyadhara et al., 2022; Vignali et al., 2022).

4
 The intensity of the applied PEF determines whether the pore formation will be
reversible or irreversible. Reversible electroporation occurs when the pore exists only
during the application of the electric field. Consequently, after the application, the
membrane returns to its original conformation. However, irreversible electroporation,
also referred to as permanent, happens when the membrane is ruptured due to the
application of the PEF (Demir, Tappi, Dymek, Rocculi, & Gómez Galindo, 2023;
Rostamabadi et al., 2023). Figure 1A shows reversible and irreversible electroporation.
Electroporation depends on the size, type and shape of the cell, membrane characteristics,
process conditions, among other factors. In general, the presence of pores increases cell
permeability through the membrane. Therefore, obtaining intracellular compounds
present in the plasma is possible (López-Cruz, Sandoval-Contreras, & Iñiguez-Moreno,
2023; Naliyadhara et al., 2022; Vignali et al., 2022). Furthermore, electroporation
compromises cellular integrity, resulting in alterations in enzymes and proteins,
culminating in microbial inactivation (Arya et al., 2023; V. Lima, Pinto, & Saraiva, 2023;
Pravallika & Chakraborty, 2022; Vignali et al., 2022).

The PEF treatment involves the application of high-voltage electric current (10 to
80 kV/cm) in short-duration intervals known as pulses, lasting approximately from
microseconds to nanoseconds (Cavalcanti et al., 2023; Ghoshal, 2023; Pravallika &
Chakraborty, 2022; Raghunath et al., 2023). Thus, the efficiency of the treatments of this
innovative technology in food and beverage quality depends on the process conditions,
such as field strength (10 - 80 kV/cm), pulse width (0.2 - 80 µs) and frequency (1 - 1500
Hz), treatment time (100 - 800 µs), membrane polarity, electrodes used, food matrix,
sample conductivity, among others (Arshad et al., 2021; Faisal Manzoor et al., 2021;
Kantala, Supasin, Intra, & Rattanadecho, 2022; L. Li, Yang, & Zhao, 2021; Mendes-
Oliveira et al., 2022; Oziembłowski, Trenka, Czaplicka, Maksimowski, & Nawirska-
Olszańska, 2022; Pravallika & Chakraborty, 2022). Figure 1B shows the PEF application
modes in a continuous and batch system. During treatment with the electric field, the
product can heat up (Vignali et al., 2022). This phenomenon is a consequence of the Joule
effect, in which heat is generated when an electric current passes through a conductor.
Yildiz, Pokhrel, Unluturk, and Barbosa-Cánovas (2019) applied PEF (35 kV/cm,
treatment time 27 µs, 155 Hz, 14 pulses) to strawberry juice and observed a temperature
increase from 22.7 to 46.0 ºC. Kantala et al. (2022) applied PEF (20 to 40 kV/cm, 10 to
50 pulses, pulse width 10 µs, 1 Hz) with different field strengths and number of pulses to
orange juice initially at 25.0 ºC and, after treatment, the temperature ranged from 26.1 to
43.6 ºC. For this reason, temperature monitoring and control measures such as the use of
cooling baths are recommended so that PEF treatment can truly be classified as a non-
thermal treatment (Pravallika & Chakraborty, 2022; Vignali et al., 2022).

Among emerging technologies, PEF and OH are techniques that involve the
application of electric current through two electrodes in a known mass or food product to
alter characteristics, achieve microbial and enzymatic inactivation, among other
consequences. The differentiation between the two techniques lies in the mode of
applying electric current and the process characteristics, specifically the temperature at
which the process occurs. PEF is a non-thermal technique, meaning that sample heating
is not desired, and thus, the sample temperature should not exceed 50 ºC. Another
difference is that the application of electric current occurs in short intervals in PEF
treatment, as mentioned earlier, in the form of pulses. On the other hand, OH is a
technique that aims to heat the sample through the Joule effect, which results from the
passage of electric current through the material or conductive medium. During ohmic
5
treatment, there are effects due to sample heating and non-thermal effects related to the
passage of electric current through the medium, such as electroporation, for example. The
sample heating promotes the inactivation of enzymes and deteriorating microorganisms,
elimination of pathogens, and other changes. For the application of PEF and OH, it is
necessary to have some equipment such as electric field generators, electrodes, pumps in
the case of continuous systems, among others. Specifically in the case of PEF, the
presence of an oscilloscope and field controller is extremely necessary to monitor the
application of the electric field in the form of pulses, i.e., discontinuously. On the other
hand, during OH application, the most essential equipment includes the power supply for
electric current, the pair of electrodes, and the treatment chamber. In PEF processing, the
process variables are field strength, frequency, pulse width, total treatment duration. In
contrast, OH presents the electrical conductivity of the sample, frequency and waveform,
heat capacity, and electrode configuration as process variables (Aghajanzadeh, Ziaiifar,
& Verkerk, 2023; Doan, Lai, & Le, 2023; Kaur, Kumar, Samota, & Lalremmawii, 2024;
Vignali et al., 2022).

3. Effects of PEF on microbiological quality

PEF technology has been investigated as an alternative for quality control and
microbiological safety in the juice and plant-based beverage industry. The exploration of
non-thermal processing alternatives aims primarily at preserving the sensory and
nutritional characteristics of the products (Bocker & Silva, 2022a; Cavalcanti et al., 2023;
Naliyadhara et al., 2022; Salehi, 2020a; Vignali et al., 2022). Therefore, PEF technology
has been studied as an alternative to conventional pasteurization and sterilization
treatments.

The mechanism of microbial inactivation associated with PEF is not completely

elucidated, however the phenomenon of electroporation that occurs in membranes is the
most widely accepted cause of cell death in microorganisms. The electropermeabilization
of cell membranes in bacteria, fungi, yeast, among others, caused by high-voltage pulses,
can lead to various effects ranging from mass transfer, such as water exudation and
intracellular compound diffusion, to impairment of cellular functions, cell lysis,
consequently resulting in cell death (Ashrafudoulla et al., 2023; Cavalcanti et al., 2023;
Duvoisin et al., 2022; Ghoshal, 2023; Pravallika & Chakraborty, 2022). Freire, Lattanzio,
Orera, Mañas, and Cebrián (2021) investigated the inactivation of Staphylococcus aureus
and the release of metal ions from the cells after applying PEF (18.0 and 25.0 kV/cm, 0.5
Hz, pulse width 4 µs, 2.59 and 5.00 kJ/kg). The authors observed inactivation ranging
between 1.0 and 3.5 logarithmic cycles of Staphylococcus aureus and the release of 98%
and 15% of potassium and magnesium ions, respectively, which constitute the cells.

Factors intrinsic to microbial cultures determine the effectiveness of PEF treatment,

including the size and geometric shape of the cells, presentation (sporulated or vegetative
form), type of microorganism, characteristics of the cell membrane, cultivation
conditions, phase of microbial growth, presence of toxic substances in the medium,
among other factors. In this context, bacteria are more resistant to PEF than molds and
yeasts due to their smaller cell size. A cell has an irregular distribution of positive and
negative charges on both sides of the membrane, resulting in a difference in electrical
potential across the cell membrane, which is called transmembrane potential. When the
6
electric field is applied, the ions are reorganized along the cell membrane, promoting
changes in the electrical potential differential. This change in electrical potential is called
induced transmembrane voltage (ITV), which, when it reaches a certain level of potential
difference, results in the phenomenon of electroporation. The ITV is directly proportional
to the applied electric field strength and the cell's radius. In this context, two cells with
different radius require different electric field intensities to reach the same ITV.
Consequently, the cell with a larger radius will require a less intense electric field
compared to the smaller cell. Another very important point is that the extracellular and
intracellular media are electrically conductive, while the cell membrane has a region
exposed to conductive media and another insulating region, thus, areas exposed to
environments with electrical conductors are more susceptible to electroporation. In this
context, the larger the area exposed to extracellular and intracellular media, the more
sensitive it is to electroporation. In addition, bacteria have a peptidoglycan layer which
makes the structure of the cell membrane more rigid. Bacterial spores, on the other hand,
are resistant to treatment because the structure of the envelope prevents electroporation.
Regarding shape, non-spherical microorganisms tend to be more resistant to PEF, once
cell orientation during field application influences the effectiveness of the treatment. This
resistance is attributed to some types of bacteria, protozoa, among others. Regarding
growth phase, cultures in the exponential phase are more sensitive to PEF compared to
the stationary phase. During the cell division process, the size of the cell and cell
membrane increases, thus the lipid bilayer becomes more prone to electroporation.
Furthermore, the presence of substances such as ethanol and acids can increase the
effectiveness of the electric field, since electroporation, even if reversible, promotes mass
transfer. Therefore, acids and other toxic substances diffuse into the intracellular
environment and compromise cellular activities (Cebrián, Mañas, & Condón, 2016;
Delso, Martínez, Cebrián, Álvarez, & Raso, 2020; Delso, Martínez, et al., 2022; El-Hag,
Jayaram, Gonzalez, & Griffiths, 2011; Freire et al., 2021; Garner, 2019; Martens et al.,
2020; Nabilah, Sitanggang, Dewanti-Hariyadi, Sugiarto, & Purnomo, 2022; Saldaña,
Monfort, Condón, Raso, & Álvarez, 2012).

Table 1 presents the impacts of PEF on microbial inactivation in fruit and vegetable
juices. In some studies, the reduction in microbial load of molds and yeasts is greater than
that of bacteria (Jin & Aboelhaggag, 2022; Jin, Aboelhaggag, & Guo, 2021; Yildiz et al.,
2019, 2021). The mechanisms behind the previously mentioned phenomenon are not
specified by Yildiz et al. (2021) and Jin and Aboelhaggag (2022). However, Jin et al.
(2021) correlate cell size with resistance to PEF, while Yildiz et al. (2019) justified this
resistance based on the morphological characteristics of different cell types. In this case,
they exemplified that larger cells of molds and yeasts are more easily permeabilized
compared to smaller cells such as bacteria (Yildiz et al., 2019).

In other studies, the reduction in bacterial microbial load was greater than the
reduction in molds and yeasts (Z. Ahmed et al., 2021; L. Li et al., 2021; Manzoor et al.,
2020). The authors of the studies did not point out a justification for the greater reduction
in the microbial load of bacteria compared to molds and yeasts. However, they proposed
techniques and processes to extend the shelf life of beverages using PEF and explained
the effects of PEF on cell structures (Z. Ahmed et al., 2021; Dziadek et al., 2019; L. Li et
al., 2021; Manzoor et al., 2020).

Delso, Berzosa, et al. (2022) discussed the phenomenon of electroporation in

microorganisms, emphasizing the dependence of the local transmembrane voltage. The
7
potential across the membrane generated by an external electric field depends on the size
of the cell and the intensity of the field. The lower reduction in bacterial count is related
to the sporulated form, as these are not affected by the PEF process. The authors found
that PEF treatment (17.5 kV/cm, 16 pulses, 10 µs, 75 Hz, 73.6 kJ/kg) in grape juice
inactivated all the microorganisms evaluated (molds, yeasts and total aerobic bacteria) to
below the detection limit, regardless of whether the grapes were treated with PEF or not.
They attributed this result to the uniform distribution of the electric field, which is related
to the use of electrodes configured in parallel. Furthermore, after PEF treatment, the
samples were initially heated up to 65 ± 1 ºC at 20 ºC. Thus, a synergistic effect of the
increase in temperature contributed to the degree of inactivation observed in the study.

Yildiz et al. (2021) compared the characteristics of strawberry juice processed by

pasteurization (TT - 72 ºC, 15 s), HIUS (55 ºC, 24 kHz, 0.29 W/mL, 120 µm, 3 min),
HPP (300 MPa, 1 min), and PEF (35 kV/cm, 155 Hz, 2 µs, 27 µs). The process conditions
for each technology were established based on the reduction of 5 log CFU/mL of
Escherichia coli O157:H7. Thus, the reduction in E. coli microbial count was the criterion
established for process equivalence. Considering the results of the microbial reduction in
the total count of aerobic and mesophilic bacteria (TMAC) and molds and yeasts (MY),
the thermal treatment applied to the strawberry juice was the most effective for
microbiological control with log reduction of 1.71 and 1.80, respectively. Processing with
HIUS and HPP showed the same reduction for MY (1.70 log reduction); however, HPP
was more efficient in reducing TMAC (1.41 log reduction). Among all observed
treatments, PEF reduced both MY and TMAC the least with 1.60 and 1.71 log reduction,
respectively; however, it is observed that the difference in microbial reduction among
different treatments is very small. Therefore, further studies using these technologies are
necessary for a more thorough evaluation concerning the microbial reduction criterion.

In summary, PEF treatment is a potential alternative for microbial inactivation in

fruit and vegetable juices since this technology has shown substantial reductions in
microbial populations. Indeed, many studies have reported PEF-induced reductions more
than 5.00 log CFU/mL for different microorganisms (Dziadek et al., 2019; Jin &
Aboelhaggag, 2022; Jin et al., 2021; Kantala et al., 2022; Mendes-Oliveira et al., 2022;
Stübler et al., 2020; Yildiz et al., 2023).

4. Effects of PEF on the physicochemical characteristics

Physicochemical parameters such as pH, titratable acidity, total soluble solids
content and color are indicators used to evaluate the quality of fruit and vegetable juices.
These characteristics have an intrinsic relationship with the sensory attributes perceived
by the consumer. Therefore, processed juices that have properties close to fresh juice in
natura are more likely to be accepted by consumers. In this context, PEF technology has
emerged as an alternative in the processing of natural juices, as it has the potential to
preserve the physicochemical characteristics inherent to juices.

4.1. Total soluble solids, titratable acidity and pH

pH, titratable acidity (TA) and total soluble solids (TSS) are physicochemical
properties often used to determine the quality of fruit and vegetable juices. Table 2 shows
the effects of PEF on pH, titratable acidity and soluble solids content. In most studies,
8
PEF did not alter the physicochemical characteristics of fruit and vegetable juices even
under conditions of high-energy application.

In general, total soluble solids refer to substances dissolved in the aqueous phase of
the evaluated sample. These substances include sugars, acids, phenolic compounds,
pectins, vitamins, salts, among others (Chavan et al., 2023; Kusumiyati, Hadiwijaya,
Putri, Mubarok, & Hamdani, 2020). In fruit and vegetable juices, the main solids present
are carbohydrates such as sucrose, glucose, and fructose. Thus, the TSS in juices is
associated with the carbohydrate content. This physicochemical property is related to the
sensory attribute of sweetness in fruit and vegetable juices (Khandpur & Gogate, 2015;
Shaik, Hamdi, & Sarbon, 2023).

Several studies have indicated that PEF did not promote significant changes in TSS
in fruit juices (Akdemir Evrendilek et al., 2021; Akdemir Evrendilek et al., 2020; Faisal
Manzoor et al., 2021; Jin & Aboelhaggag, 2022; Jin et al., 2021; Kantala et al., 2022; L.
Li et al., 2021; Mendes-Oliveira et al., 2022; Rahaman et al., 2020; Yildiz et al., 2021).
Some studies reported an increase in TSS after processing with PEF (Atencio et al., 2022;
Buitimea-Cantúa et al., 2022). Atencio et al. (2022) studied the effects of PEF (1.01
kV/cm, pulse width 80 µs, 9 kJ/kg) on pumpkin juice and observed an increase in TSS
from 2.54 to 2.80 ºBrix. Buitimea-Cantúa et al. (2022) applied PEF (11.3 to 23.3 kV/cm,
pulse width 25 µs, 100 pulses, 19.7 to 168.4 kJ/L) to raspberry juice. The results show an
increase in TSS from 7.5 to 8.5 ºBrix in all PEF treatments. As mentioned by Atencio et
al. (2022) and Buitimea-Cantúa et al. (2022), the increase in total soluble solids occurs as
a result of the release of carbohydrates from inside the cells into the aqueous medium of
the juice, in addition to the action of enzymes responsible for producing these
carbohydrates.

However, some studies observed a reduction in TSS values (T. Ahmed et al., 2022;
Rahaman et al., 2020; Wibowo et al., 2019). T. Ahmed et al. (2022) applied PEF (5
kV/cm, treatment time 20 min) to pineapple juice and observed a reduction in TSS from
12.06 to 11.03 ºBrix. Wibowo et al. (2019) evaluated the effects of PEF treatment (12.5
kV/cm, pulse width 2 µs, 62 and 94 Hz, 76.4 and 132.5 kJ/L) on cloudy apple juice,
resulting in the least intense treatment of 76.4 kJ/L reducing the soluble solids content
from 13.08 to 12.78 ºBrix. Electroporation caused by PEF allows mass transfer across
cell membranes. Thus, during the application of PEF, the soluble solids may have diffused
into the intracellular environment, resulting in a reduction in the substances present in the
aqueous phase. In this sense, before the application of PEF, the inside of the cells had a
lower concentration of solids than the juice, thus resulting in the mass transfer of sugars
from the juice to the inside of the cell structures present in the medium. This explanation
was presented by T. Ahmed et al. (2022) to account for the reduction in TSS in pineapple
juice treated with an electric field.

The organic acids present in fruits and derived products play an essential role in the
perception of sour taste by consumers and in extending the shelf life of these foods
(Asencio, Serrano, García-Martínez, & Pretel, 2018; Batista-Silva et al., 2018;
Chahardoli, Jalilian, Memariani, Farzaei, & Shokoohinia, 2020). Moreover, organic acids
constitute a significant portion of the soluble solids in fruits, accounting for
approximately 1% of the total fruit mass (Salehi, 2020b). Therefore, the content of
organic acids directly influences the stability, flavor, acceptance, and functionality of fruit
and vegetable juices. The main acids present in fruit and vegetable juices are citric acid,
9
tartaric acid and malic acid. The concentration of acids in a product can be determined by
titratable acidity.

In several studies, PEF treatment shown to have no significant effects on TA

(Akdemir Evrendilek et al., 2021; Jin & Aboelhaggag, 2022; Jin et al., 2021; Mendes-
Oliveira et al., 2022; Wibowo et al., 2019; Yildiz et al., 2021).

Oziembłowski et al. (2022) obtained divergent results from the previously

mentioned findings when they applied PEF to chokeberry juice. The fruit used to prepare
the juice was obtained from six different farms in Poland. Of the six juice samples, only
one showed an increase in TA from 0.98 to 1.04 g of malic acid/100 mL of juice.
Considering that the application of the electric field leads to increased molecular mobility
of phytochemical compounds due to electroporation, the rise in TA might result from the
mass transfer of acids from the interior of cellular structures and organelles into the
aqueous medium.

pH is a physicochemical property that applies a logarithmic scale to determine the

degree of acidity of a sample, thus allowing the sample to be classified as acidic, neutral
or alkaline. The pH is determined based on the concentration of hydrogen cations (H+)
present in the medium. In the context of vegetable juices and drinks, pH is affected by the
content of organic acids naturally present in the raw material. The main acids present in
fruit and vegetable juices are citric acid, tartaric acid and malic acid, which are considered
weak acids. As such, the hydrogen cations of these acids do not dissociate completely in
aqueous solution, resulting in a lower contribution of these acids to the pH when
compared to strong acids.

The impact of PEF on the pH of fruit and vegetable juices is not consistently
reported in research on the subject. Some studies have indicated that PEF treatment did
not result in significant changes in pH (Akdemir Evrendilek et al., 2021; Akdemir
Evrendilek et al., 2020; Jin & Aboelhaggag, 2022; Jin et al., 2021; Kantala et al., 2022;
L. Li et al., 2021; Mendes-Oliveira et al., 2022; Yildiz et al., 2021).

Some studies have identified an increase in pH after PEF treatment (T. Ahmed et
al., 2022; Buitimea-Cantúa et al., 2022; Wibowo et al., 2019). However, the increase in
pH implies a reduction in the concentration of ionic hydrogen in the medium. This could
result from the mass transfer of organic acids from the continuous phase into the cellular
structures.

Considering that PEF enhances substance mobility through electroporation, another

possibility is the reduction in pH, which might be related to the release of organic acids
from the intracellular medium into the extracellular environment. The increased content
of organic acids elevates the concentration of hydrogen ions, consequently lowering pH
values. Atencio et al. (2022) applied PEF (1.01 kV/cm, 40 pulses, 80 μs, 9 kJ/kg) to
pumpkin juice and observed a reduction in pH from 7.03 to 6.73. Rahaman et al. (2020)
treated apricot juice with PEF (7 and 14 kV/cm, treatment time 500 µs, 1 kHz) and the
pH decreased from 2.99 to 2.98 and 2.92 for the 7 and 14 kV/cm treatments, respectively.

Buitimea-Cantúa et al. (2022) investigated pH variations in raspberry juice resulting

from PEF treatment (11.3 to 23.3 kV/cm, pulse width 25 μs, 100 pulses, 19.7 to 168.4
kJ/L) at different frequencies (100, 200, and 500 Hz). All juice samples subjected to PEF

10
treatment exhibited a higher pH than the untreated sample. Remarkably, treatments at
frequencies of 100 Hz (19.7 and 79.7 kJ/L) and 500 Hz (113.8 kJ/L) resulted in the highest
pH values. Concurrently with the pH increase, the results showed a reduction in TA.
Therefore, the variations observed in pH and TA may be associated with the trapping of
these substances in the raspberry cell matrix after electroporation caused by PEF. Sensors
used for pH determination, for instance, quantify ions present in the aqueous medium.
Therefore, when compounds migrate into the cell interior, fluctuations in the equipment's
measured readings might be noticed. Similar reasoning applies to TA, the analysis is
based on the amount of basic ions needed to neutralize the acid content present in the
aqueous medium of the sample. In this context, acids trapped within cellular structures
do not react with added base for neutralization.

In this context, the application of PEF induces little or no significant change in the
physicochemical characteristics of vegetable and fruit juices. The TSS, TA, and pH are
quality parameters in the juice industry, and thus, preserving these properties is ideal
when the goal is the development of a new process for the chemical and kinetic
stabilization of products.

4.2. Color

Color has a significant influence on consumer acceptance of beverages. Fruit and

vegetable juices that are closer in color to the natural color of fresh fruit tend to be more
acceptable. This preference is attributed to the association between color and the freshness
of the juice (Jin et al., 2021). The main pigments attributed to the colors of fruit and
vegetable juices are anthocyanins, betalains, carotenoids and chlorophylls, which are
responsible for the red-blue-purple, red, yellow-orange-red and green coloring,
respectively (Rodriguez-Amaya, 2016). The coloring and stability of each pigment are
intrinsically related to the environmental conditions to which they are subjected (Bocker
& Silva, 2022b). Therefore, it is crucial to minimize changes in the color of fruit and
vegetable juices during processing.

The instrumental assessment of juice color is based on the CIELAB system, a color
model representing hues as perceived by the human eye. Ele é amplamente utilizado na
indústria, design gráfico, ciência dos materiais e em muitas outras áreas. It is widely used
in industry, graphic design, materials science and many other areas. The LAB system is
made up of three main components: L* (lightness), a* (red-green component) and b*
(blue-yellow component). The coordination of these values describes a specific color. The
coordination of these values describes a specific color. Thus, the total color difference
(ΔE) is derived from the comparison between the processed sample and the untreated
sample. According to Rahaman et al. (2020), when ΔE is less than 2, changes in color are
not noticeable by consumers.

As mentioned previously, each group of natural pigments contributes to different

color ranges. The color stability of these pigments depends on pH, temperature, exposure
to light and heating (Ngamwonglumlert, Devahastin, & Chiewchan, 2017). Morever,
other factors can contribute to the color of fruit and vegetable juices. Changes in the
concentration of natural pigments in the medium because of degradation by heating,
variations in pH and exposure to light. Phenolic compounds exposed to oxidative
11
enzymes generate dark pigments that darken the products. In processes that involve
heating the raw material, it is possible for the Maillard reaction to occur, resulting in the
formation of melonoidins, which favors the browning of the product. Thus, fruit and
vegetable juices can contain multiple groups of pigments, and the color of the product
depends on the interaction between all the substances present in the medium.

Table 3 shows studies that evaluated the effects of the PEF process on the color
variation of fruit and vegetable juices. As reported by several studies, PEF does not alter
or promotes subtle changes in the color of vegetable juices (Akdemir Evrendilek et al.,
2020; Faisal Manzoor et al., 2021; Jin et al., 2021; Kantala et al., 2022; Rahaman et al.,
2020). PEF treatment is a technology that can be applied in a non-thermal way, reducing
the chances of the Maillard reaction occurring. Additionally, it can promote a reduction
in polyphenol oxidase activity, resulting in few changes due to enzymatic browning. In
another way, PEF can alter the concentration of each group of pigments in the medium
because of the electropermeabilization of cell membranes. These changes in the
concentration of pigments were not very significant, as can be seen in most of the studies
that were reviewed.

T. Ahmed et al. (2022) observed that PEF treatment induced changes in color in
pineapple juice, where the reduction in the "L" value was associated with non-enzymatic
browning reactions, degradation of carotenoids, anthocyanins, vitamins, and other juice
components. Faisal Manzoor et al. (2021) applied PEF (9 kV/cm, pulse width 80 µs,
treatment time 335 µs, 1 kHz), HIUS (180 W, 40 kHz, 21 min, 30 ºC) and the combination
of these two technologies under the conditions described above to spinach juice. The
sample treated with PEF showed the lowest color variation (1.73) followed by HIUS
(2.42), while the combination of technologies (PEF + HIUS) varied the color by 3.31.
According to the authors, the color change seen after PEF treatment was related to the
phenomenon of electroporation, which released oxidative enzymes and other components
into the medium, resulting in a color change in the juice samples.

In the context of applying PEF in the processing of vegetable and fruit juices, color
changes occur at little relevant and imperceptible intensities to the consumer.
Furthermore, among emerging technologies, such as HIUS, HPP, microwave heating, and
thermal treatments, the PEF is the alternative that induces the least color alterations
depending on the employed process conditions.

5. Effects of PEF on enzymatic inactivation

Various enzymes naturally exist in plant tissues, serving vital functions for their
survival. However, enzyme activity can be a problem during the storage of products of
plant origin. Sedimentation of solid particles, browning and the development of off-
flavors are consequences of the action of enzymes in fruit and vegetable juices (Basak &
Chakraborty, 2022; Singh et al., 2022; Szczepańska, Barba, Skąpska, & Marszałek,
2022). Monitoring enzyme activity in fruit and vegetable juices is crucial when the aim
is to preserve the sensory, nutritional and functional quality of the product for as long as
possible.

12
 The main enzymes found in juices are polyphenol oxidase (PPO), peroxidase
(POD) and pectin methylesterase (PME). POD and PPO cause changes in the color of
vegetable and fruit juices because of the oxidation of phenolic compounds, thus
promoting the browning of the product. PME functions by de-esterifying the methoxyl
groups of pectin, leading to the loss of cloud stability. This occurs because the methyl
groups of pectins prevent polymerization and gelatinization reactions. Consequently,
through the reduction of the degree of methoxylation, cationic ions, such as calcium, can
interact with the pectin lacking the methyl group, resulting in the formation of insoluble
pectate. Because of the reduced solubility of these polymers, fruit and vegetable juices
experience a loss of cloud stability and phase separation in the product. Furthermore, the
action of PME can alter other quality parameters of juices, such as viscosity, color, and
flavor. Considering these considerations, the inactivation of PME becomes a decisive
factor for preserving the characteristics of vegetable and fruit juices, resulting in kinetic
and chemical stability for the product (Aghajanzadeh & Ziaiifar, 2018; Qian et al., 2021;
Wang et al., 2022). These enzymes are naturally present in the plant tissues of fruits
commonly used for juice production (Z. Ahmed et al., 2021; Manzoor et al., 2020).

Table 4 presents a compilation of studies investigating enzyme inactivation caused

by PEF in fruit and vegetable juices. Electric field processing is a viable alternative for
quality control in terms of enzyme inactivation. Faisal Manzoor et al. (2021) evaluated
the enzymatic inactivation of PPO and POD in spinach juice treated with PEF (9 kV/cm,
pulse width 80 µs, treatment time 335 µs, 1 kHz). The results indicated a reduction of
44.00% and 43.52% in PPO and POD activity, respectively. Z. Ahmed et al. (2021)
applied PEF (9 kV/cm, pulse width 80 µs, treatment time 335 µs, 1 kHz) to wheat plant
juice and assessed enzymatic activity, showing a reduction of 35.63% and 38.70% in the
activity of POD and PPO enzymes, respectively. Regarding the effects of PEF on PME
inactivation, Buitimea-Cantúa et al. (2022) treated raspberry juice under conditions of
11.3 to 23.3 kV/cm, pulse width 25 µs, 100 pulses, 19.7 to 168.4 kJ/L. The highest PME
inhibition was observed in the treatment of 168.4 kJ/L, with a 19% activity inhibition,
followed by treatments of 79.7 and 113.8 kJ/L, both resulting in 13% inhibitions.

Wibowo et al. (2019) investigated the impacts of PEF (12.5 kV/cm, pulse width 2
µs, 62 and 94 Hz, 76.4 and 132.5 kJ/L) on enzymes in cloudy apple juice. The PEF
treatment with 76.4 kJ/L reduced approximately 36%, 49%, and 50% of PPO, POD, and
PME activity, respectively, while the treatment with 132.5 kJ/L reduced the activity of
both enzymes by over 90%. In this study, the juice temperature after PEF treatments was
60°C and 73°C for energy densities of 76.4 and 132.5 kJ/L, respectively. This study also
compared PEF processing with thermal treatments TP1 (72°C/15 s) and TP2 (85°C/30 s).
The reductions in the activity of enzymes POD, PPO, and PME were similar to the
thermal treatments. Therefore, sample heating may have a synergistic effect when
combined with PEF application concerning the reduction of activity in these enzymes.

The mechanism of enzyme inactivation with PEF treatment has not yet been fully
elucidated. Some studies have proposed/suggested the mechanisms of enzyme
inactivation by the electric field. Faisal Manzoor et al. (2021) proposed that PEF induces
protein polarization, leading to changes in structural conformation and consequently
resulting in protein denaturation. Z. Ahmed et al. (2021) indicated that the approach
attributed to non-thermal technologies generally involves changes in the enzymes' tertiary
structure, which corresponds to their three-dimensional conformation. Zhao and Yang
(2010) investigated the inactivation and conformational changes of lysozymes under
13
stress caused by PEF (3.5 kV/cm, 300 to 1200 µs), which displayed alterations in the
secondary structure. Some segments that originally exhibited an α-helix conformation
shifted to β-sheets, and furthermore, other sections showed a random conformation that
was not observed before PEF treatment. Also in this study, the authors also compared
enzymatic inactivation caused by PEF and thermal treatment (100°C/30 s). The structural
changes in the enzyme treated by heating were associated with alterations in the protein's
tertiary structure without changes in secondary structures. This evidence supports the idea
that the mechanisms of action for distinct processing methods differ in enzymatic
inactivation. In this context, the changes in conformation in both secondary and tertiary
structures lead to alterations in the active sites of enzymes, inhibiting the interaction
between substrate and protein, consequently reducing the activity of these enzymes.

According to Premjit and Sruthi (2023), the energy supplied by the electric field is
absorbed by the polar groups of proteins, leading to the formation of free radicals,
aggregation, and unfolding of the protein structure. Consequently, these free radicals
disrupt the intermolecular interactions of protein (such as electrostatic attraction,
hydrogen bonding, van der Waals forces, among others), ultimately altering the structural
conformation and denaturing the proteins. These authors further suggested that the initial
energy from PEF leads to changes in the protein structure, exposing hydrophobic amino
acids to the aqueous environment, thus altering the conformation of enzymes in three-
dimensional space, ultimately promoting protein denaturation.

Similar to the phenomenon of electroporation in the cell membrane, PEF causes a

redistribution of charges in the enzyme structure. Consequently, regions of electrostatic
attraction and repulsion are altered. This results in modifications in secondary structures
(α-helix and β-sheet) as well as tertiary structures. In this context, after their structure is
altered by the electric field, considering that the function and activity of proteins and
enzymes depend on their structure, these macromolecules lose their original biological
function.

On the other hand, PEF can increase enzymatic activity by facilitating the release
of enzymes from the intracellular to the extracellular environment through the process of
electroporation, as observed by Tian et al. (2018). Freire et al. (2021) applied PEF to
Staphylococcus aureus aiming to obtain intracellular compounds (18.0 and 25.0 kV/cm,
0.5 Hz, pulse width 4 µs, treatment time 20 and 400 µs, 2.59 and 5.00 kJ/kg). Treatments
at 18 kV/cm for 400 µs and 25 kV/cm for 20 µs showed similar levels of electroporation
but different levels of microbial inactivation. Considering this result of inactivation, the
quantity of proteins released from the interior of cells by these treatments was compared,
and the treatment with 18 kV/cm for 400 µs exhibited a significantly higher release
compared to 25 kV/cm for 20 µs. Regarding the molecular weight of these proteins, the
bands with higher occurrence were at 8.5 and 6.7 kDa, respectively. Therefore, PEF
demonstrates potential for releasing proteins and enzymes from cellular matrices.
However, further studies are necessary to comprehend how different process variables,
such as pore and protein sizes, and other factors, influence this phenomenon.

To conclude, although the mechanisms of enzymatic inactivation caused by PEF

are not evident. Some studies have reported a reduction in the activity of deteriorating
enzymes due to PEF treatment. Different enzymes, such as PPO, POD, and PME, showed
varying levels of inactivation when the same process conditions were applied.
Furthermore, increasing the intensity of the electric field treatment reduces enzymatic
14
activity because higher energy levels induce more changes to the enzyme structure. In
this context, the inactivation of PPO and POD preserves the nutritional and sensory
qualities, preventing browning and flavor alterations in the juices. Simultaneously, the
inactivation of PME by PEF contributes to greater cloud stability and reduced insoluble
pectin, consequently minimizing the formation of precipitates in vegetable and fruit
juices.

6. Effects of PEF on functional and nutritional properties

Vegetables, fruits and derived products, such as juices, exhibit a variety of bioactive
compounds produced due to cellular secondary metabolism, including phenolic
compounds, proteins, lipids, terpenes, organic acids, carotenoids, polysaccharides, among
others. Bioactive compounds can maintain biological activities through different
mechanisms of action, such as antioxidant activity, anti-inflammatory, neuro-, cardio-
and hepato-protective effects, anti-cancer, anti-obesity, anti-diabetics, improvements in
the digestive and immune systems, among others. Vegetable and fruit juices are excellent
carrier systems or vehicles for nutrients in the body. Furthermore, they are also recognized
as products that provide pleasurable sensory experiences, offer ease of consumption,
present a wide variety of fruits and vegetables for obtaining juices, providing a variety of
bioactive compounds, easy transportation and storage, among others factors (Barros et
al., 2023; Gupta et al., 2023; Olías, Delgado-Andrade, Padial, Marín-Manzano, &
Clemente, 2023; Patil, Crosby, Byrne, & Hirschi, 2014; Todaro et al., 2023; Wu et al.,
2023).

6.1. Phenolic compounds

Phenolic compounds are substances derived from the secondary metabolism of

plants, thus forming a group of thousands of components. Chemically, the term "phenolic
compound" applies when the chemical structure contains one or more aromatic rings with
at least one hydroxyl group attached to the ring. The main families of phenolic compounds
are flavonoids, non-flavonoids (tannins, stilbenes, and lignans), and phenolic acids
(Barkaoui, Madureira, Boudhrioua, & Cabo Verde, 2023; Gupta et al., 2023; Mir-Cerdà,
Nuñez, Granados, Sentellas, & Saurina, 2023). Phenolic compounds exhibit various
beneficial nutraceutical effects for the human body, acting as antioxidants, anticancer
agents, anti-inflammatory compounds, antidiabetics, and as prophylaxis for
cardiovascular, neurological, and age-related diseases (Barkaoui et al., 2023; Mir-Cerdà
et al., 2023; Parohan et al., 2019). Additionally, these compounds contribute to the
neutralization and scavenging of free radicals and act against heavy metal toxicity through
chelation capacity (Mir-Cerdà et al., 2023). Other factors, such as enzymatic reactions –
like the action of PPO – polymerization with proteins, and excessively intense
processing, can cause a reduction in the content of phenolic compounds in foods
(Panigrahi, Mishra, & De, 2023).

Vegetable and fruit juices contain the phenolic compounds originally present in
fruits. In addition to their high nutritional value, antioxidant activity, and various health
benefits, phenolic compounds, specifically flavonoids and anthocyanins, can contribute
to the sensory characteristics of juices by enhancing the intensity of the final color of the
product. These substances are pigments found in plants, fruits, and flowers, and they play
a role in improving the color intensity of the final juice product (Buitimea-Cantúa et al.,
15
2022). Therefore, preserving phenolic compounds in juices during processing allows for
the creation of juices that not only offer nutritional value but also contribute to health due
to their nutraceutical effects. In this context, PEF technology emerges as a promising non-
thermal technique for preserving the nutraceutical properties of vegetable and fruit juices
by maintaining phenolic compounds.

Table 5 presents studies published in the literature that investigate the impact of
PEF on the total phenolic content in fruit and vegetable juice beverages. Several studies
have observed that this technology does not modify the concentration of phenolic
compounds in the juice samples treated with PEF (Akdemir Evrendilek et al., 2021;
Akdemir Evrendilek et al., 2020; Dziadek et al., 2019; Jin & Aboelhaggag, 2022; Jin et
al., 2021; Oziembłowski et al., 2022; Yildiz et al., 2021). The results obtained in these
studies support the hypothesis that PEF is a promising alternative for juice processing
without compromising beneficial phytochemical compounds for health.

Some studies indicate an increase in the content of phenolic compounds present in

the medium after applying PEF (Z. Ahmed et al., 2021; Buitimea-Cantúa et al., 2022;
Faisal Manzoor et al., 2021; Rahaman et al., 2020; Yildiz et al., 2021). The results
obtained in these studies may be associated with the phenomenon of electroporation,
which enables the mobility of molecules through the cell membrane. Phenolic compounds
are produced and contained within cellular organelles. During PEF application, these
compounds traverse the membrane due to the formed pores, becoming available in the
liquid medium. Consequently, they are accounted for in the analyses characterizing the
content of phenolic compounds. In this sense, the analysis to determine the content of
phenolic compounds can quantify these compounds that were previously trapped within
the cellular matrix. Additionally, the increase in content of phenolic compounds is
associated with an increase in antioxidant capacity, as observed in several studies (Z.
Ahmed et al., 2021; Faisal Manzoor et al., 2021; Rahaman et al., 2020; Yildiz et al., 2021).
Phenolic compounds exhibit antioxidant properties, thus, the release of these compounds
into the aqueous medium enhances the antioxidant capacity of juice samples.

However, certain studies have reported a decline in the level of phenolic compounds
after PEF application (T. Ahmed et al., 2022; Jin et al., 2021). The reduction in phenolic
compounds might result from their degradation during processing. Sample heating due to
the Joule effect could induce the degradation of these compounds. These substances are
sensitive to temperature and can degrade under conditions of moderate and severe thermal
treatment (Danişman, Arslan, & Toklucu, 2015; El Hosry et al., 2023; Yıkmış, Tokatlı
Demirok, Levent, & Apaydın, 2023). Other reasons for the reduction in the content of
phenolic compounds include their oxidation due to the possible formation of free radicals
during PEF processing and polymerization with proteins (T. Ahmed et al., 2022). Jin et
al. (2021) investigated the impact of PEF treatment (19, 23, and 30 kV/cm, pulse width 2
μs, treatment time 181 μs, 1.25 kHz) on the phenolic content of apple juice. The results
indicated that the PEF treatment at 19 kV/cm caused a decrease in the quantity of phenolic
compounds, whereas the 23 kV/cm treatment showed no significant difference compared
to the untreated sample. This outcome illustrates that process conditions directly influence
the concentration of phenolic compounds in the juice. Furthermore, considering the
effects of PEF on phenolic compounds and cellular membranes, the treatment at 19
kV/cm resulted in the degradation of phenolic compounds and a relatively insignificant
release of intracellular content into the extracellular medium. Conversely, the treatment
at 23 kV/cm degraded these substances and facilitated their diffusion into the extracellular
16
medium through electroporation, thereby not significantly altering the content of phenolic
compounds when compared to the untreated sample. The antioxidant effect of juices is
related to the presence and concentration of phenolic compounds. Some studies evaluated
the antioxidant capacity after PEF application and reported an increase in this property
(T. Ahmed et al., 2022; Z. Ahmed et al., 2021; Akdemir Evrendilek et al., 2021; Akdemir
Evrendilek et al., 2020; Faisal Manzoor et al., 2021; Rahaman et al., 2020; Stübler et al.,
2020; Yildiz et al., 2021). Antioxidants compounds work to reduce free radicals, thus
preventing oxidative stress in the human body. Reactions involving free radicals cause
cellular alterations, damaging cell structures and compromising cellular function,
resulting in oxidative stress. This stress can lead to the development of chronic diseases
such as diabetes, cancer, cardiovascular diseases, among others (El Hosry et al., 2023;
Okwunodulu, Obioma, Okwunodulu, Ndife, & Wabali, 2023).

Dziadek et al. (2019) and Oziembłowski et al. (2022) evaluated phenolic

compounds and antioxidant activity in apple and aronia juice treated with PEF (1st study
– 30 kV/cm, 200 to 400 pulses, 0.033 Hz | 2nd study – 20 kV/cm, 300 pulses, pulse width
0.17 μs, 0.066 Hz), respectively. The observed reduction in antioxidant activity was not
linked to a decrease in content of phenolic compounds, as the concentrations of these
substances showed no significant changes compared to the untreated sample. Therefore,
other substances with antioxidant activity might have degraded during PEF processing.

Given this, studies have reported that PEF technology does not alter the content of
phenolic compounds. Moreover, there are studies that have shown an increase in the
content of phenolic compounds because of the PEF technology application, which is
associated with the electroporation phenomenon. Therefore, this innovative technology
does not possess a severity level that compromises the content of these bioactive
compounds present in vegetable and fruit juices.

6.2. Flavonoids and anthocyanins

Flavonoids constitute the most numerous groups of phenolic compounds. The

chemical structure of these substances consists of two aromatic rings linked by a
heterocyclic oxygen atom. Within this group, there are different families based on the
substitutions of the heterocycle and degree of oxidation (Q. Li et al., 2023; Mir-Cerdà et
al., 2023). The subcategories of flavonoids include catechins, flavones, anthocyanidins,
flavonols, flavanones, and isoflavones (Q. Li et al., 2023).

Anthocyanins are the primary flavonoids found in fruit and vegetable juices. As
water-soluble pigments, they are primarily responsible for the blue, red, and purple
coloration of these products. These pigments belong to the group of polyphenolic
flavonoids and are considered part of the anthocyanidin family. They exhibit antioxidant
capacity and are found in fruits, vegetables, and flowers (Chen et al., 2022; Lepaus et al.,
2023). Anthocyanins are polyphenols capable of protecting the human body from chronic
diseases such as diabetes, cancer, atherosclerosis, and cardiovascular diseases
(Daneshzad, Shab-Bidar, Mohammadpour, & Djafarian, 2019).

In purified form, flavonoids and anthocyanins exhibit limited stability, primarily

due to their susceptibility to nucleophilic attacks. Factors such as temperature, pH, the
presence of metals, enzymes, oxygen, light, among others, affect the stability of
anthocyanins (Chen et al., 2022; Skrt et al., 2022). The degradation of anthocyanins due
17
to thermal treatment generally involves deglycosylation reactions and/or opening of
aromatic rings, along with increased oxidation rates, resulting in the formation of
compounds with a single aromatic ring containing hydroxyls, aldehyde groups, methoxy,
or carboxyls. Thermal processing-induced degradation depends on various variables such
as the type of heating, duration, nature of the anthocyanins, and food matrix (Chen et al.,
2022). The degradation of pomegranate juice anthocyanins due to thermal treatment (80
ºC/10 min) was investigated by Skrt et al. (2022). The authors observed that after the
thermal treatment, the degradation of cyanidin-3,5-O-di-glucoside, delphinidin-3-O-
glucoside, and cyanidin-3-O-glucoside was approximately 30%. After 5 months of
storage, this value reached 90% for delphinidin-3-O-glucoside and cyanidin-3-O-
glucoside, and 65% and 75% for delphinidin-3,5-O-di-glucoside and cyanidin-3,5-O-di-
glucoside, respectively.

Therefore, the development of processes that do not compromise the functional and
sensory properties of fruit and vegetable juices is one of the major challenges in the
beverage manufacturing sector. In this regard, PEF is a non-thermal technology that
appears to be a promising alternative to meet this demand. Several studies have
investigated the impact of PEF treatment on the content of flavonoids and anthocyanins
in fruit and vegetable juices. Several studies have assessed the use of PEF technology in
processing fruit and vegetable juices, observing the preservation of the content of these
substances, resulting in the conservation of the nutraceutical properties in the beverage
(T. Ahmed et al., 2022; Akdemir Evrendilek et al., 2021; Akdemir Evrendilek et al.,
2020). Furthermore, other studies present a beneficial effect of PEF on the concentration
of these substances. Faisal Manzoor et al. (2021) reported an increase of 5.81% and 8.23%
in the content of flavonoids and anthocyanins after the application of PEF (9 kV/cm, pulse
width 80 µs, treatment time 335 µs, 1 kHz) in spinach juice. Rahaman et al. (2020)
investigated the effects of PEF (7 and 14 kV/cm, treatment time 500 µs, 1 kHz) in apricot
juice and observed an increase of 24.21% and 83.32% in the flavonoid content.

Yildiz et al. (2021) applied PEF (35 kV/cm, pulse width 2 µs, 155 Hz, treatment
time 27 µs) to strawberry juice, evaluating the anthocyanin content. The results showed
a 17.12% increase in the content of these substances. The increase in concentration of
these substances represents an enhancement in the functional and nutraceutical properties
of the juices. Additionally, alterations in the anthocyanin content might influence the
color of the beverage compared to the untreated sample. Faisal Manzoor et al. (2021) also
evaluated the color parameters of spinach juice after the PEF treatment. The values of
"L", "a", "hue (saturation)", and "chroma" were altered by the electric field treatment
compared to the untreated sample. The pores generated by PEF enable the diffusion of
substances from the intracellular to the extracellular environment. Consequently,
flavonoids and anthocyanins present within plant cells become available in the juice,
resulting in color changes.

In summary, although anthocyanins and flavonoids are substances sensitive to

oxygen, light, high temperatures, and other factors, the application of PEF did not
compromise the content of these substances in fruit and vegetable juices, as reported in
the studies. Thus, the bioactive properties and sensory characteristics, such as color, are
preserved in juices treated with the electric field.

18
7. Severity level of PEF treatment

Ascorbic acid, also known as vitamin C, is a water-soluble substance commonly

found in vegetables and fruits such as spinach, broccoli, tomatoes, oranges, lemons
among others (Phillips et al., 2018; Sir Elkhatim, Elagib, & Hassan, 2018). In the human
body, this vitamin acts as an enzymatic cofactor responsible for the biosynthesis of
regulatory molecules, preventing scurvy, reducing oxidative stress, among other
functions (Fenech, Amaya, Valpuesta, & Botella, 2019). The human body cannot
synthesize the molecule of ascorbic acid. Therefore, this vitamin needs to be obtained
through diet. The average daily recommended intake of ascorbic acid is 90 to 100 mg.
For scurvy prevention, an approximate dose of 45 mg/day is recommended, while for
health optimization, it is around 200 mg/day (Carr & Lykkesfeldt, 2021).

The ascorbic acid is sensitive to different conditions during food processing and
storage. In the processing stage, moderate to high temperatures, pressure, and shear
conditions can cause the degradation of this vitamin. Meanwhile, during storage, factors
such as exposure to oxygen, light, temperature variations, among others, are crucial for
preserving ascorbic acid. Therefore, the consumption of fresh fruits and fruit juices tends
to present higher levels of ascorbic acid when compared to the processed version of these
products (Patil et al., 2014; Phillips et al., 2018). Considering the sensitivity of this
substance to adverse conditions, the ascorbic acid content has been utilized as a parameter
to evaluate the severity of food processing (Basak, Shaik, & Chakraborty, 2023).

Studies have investigated the effects of PEF treatment on the ascorbic acid content.
Some studies suggest that PEF treatment does not alter the ascorbic acid content in fruit
and vegetable juices, indicating the capacity of technology to preserve this substance
(Akdemir Evrendilek et al., 2020; Dziadek et al., 2019; Jin & Aboelhaggag, 2022;
Mendes-Oliveira et al., 2022). As a result, the electric field processing conditions in these
studies did not compromise the chemical stability of ascorbic acid and likely other
substances sensitive to food processing, thereby preserving the nutritional properties of
the evaluated juices.

In other studies, the ascorbic acid content increased after the PEF treatment
(Buitimea-Cantúa et al., 2022; Faisal Manzoor et al., 2021). This could be a result of mass
transfer induced by electroporation from within the cells into the aqueous medium of the
juice, making ascorbic acid more available in the liquid medium and thus quantifiable by
detection methodologies.

In another hand, other studies indicate a decreasing in this compound as a

consequence of PEF processing (Etzbach et al., 2020; Kantala et al., 2022; Wibowo et al.,
2019). In this cases, the electric field processing parameters used were excessively
intense, leading to the degradation of ascorbic acid molecules. Kantala et al. (2022)
evaluated the quality parameters of orange juice treated with PEF in comparison to
thermal treatment. After the electric field processing at different field intensities (20, 30,
and 40 kV/cm, 10 to 50 pulses, pulse width of 10 µs, 1 Hz, treatment time of 100 to 500
µs), an increase in the sample temperature was observed ranging from 26.1 to 43.6 ºC,
compared to the initial ambient temperature of 25 ºC. The thermally processed sample
(68.2 ºC/30 min) showed an 18.42% reduction in ascorbic acid content, while the PEF-
treated juice showed a 7.89% reduction. Among the PEF treatments, there was no
significant difference in the ascorbic acid content. However, there was a significant
19
difference observed between the untreated sample, thermally treated sample, and PEF-
treated sample. In this case, the reduction in the ascorbic acid content during the PEF
treatment could be attributed to the rise in temperature during the field application.
Notably, the decrease in the concentration of this compound through PEF was less intense
compared to the thermal treatment. In the study conducted by Etzbach et al. (2020), the
characteristics of orange juice treated with non-thermal and thermal technologies were
evaluated. The sample's inlet and outlet temperatures during PEF treatment (12.7 kV/cm)
were 40.4 and 62.9 ºC, respectively. Furthermore, samples were thermally treated with
mild pasteurization (72.5 ºC/30 s) and conventional pasteurization (92.2 ºC/31 s). The
samples processed by PEF, mild pasteurization, and conventional pasteurization showed
reductions of 4.28%, 3.95%, and 2.64% in the ascorbic acid content, respectively.
Consequently, the PEF treatment exhibited the greatest reduction in the substance's
content, showing no significant difference when compared to mild pasteurization.
Meanwhile, conventional pasteurization demonstrated a smaller reduction in the ascorbic
acid content, thus presenting higher levels than the PEF treatment. In both previously
mentioned studies, a decrease in the content of this compound and an increase in the
sample temperature during PEF processing were observed. In the second case
specifically, the heating of the samples was more intense, reaching 62.9 ºC and estimated
to have reached 69.0 ºC after treatment. Temperature is a crucial factor in the degradation
of ascorbic acid, thus, the significant rise in temperature during the application of a non-
thermal technology may contribute to the reduction in the content of this substance.

In the study carried out by Wibowo et al. (2019), PEF was applied to apple juice
under conditions of 12.5 kV/cm, pulse width of 2 µs, frequencies of 62 and 94 Hz, at
energy levels of 76.4 and 132.5 kJ/L. Treatment with 76.4 kJ/L did not alter the ascorbic
acid content of the apple juice compared to the untreated sample. However, treatment
with 132.5 kJ/L resulted in a 10% reduction in the ascorbic acid content. Thus, the energy
density of the PEF treatment influenced the extent of degradation of this compound.

Therefore, the application of PEF can affect the ascorbic acid content in juices both
positively and negatively. The increase and preservation of ascorbic acid content after
PEF treatment indicate the occurrence of the electroporation phenomenon and the
relatively low severity of this technology. On the other hand, the reduction in the content
of this substance due to PEF treatment indicates intense process conditions and/or an
increase in the sample temperature during treatment, resulting in a decrease in the
ascorbic acid content.

8. Impact of PEF on sensory characteristics

The new consumer market demands have driven fruit and vegetable juice industries
to produce microbiologically safe products without toxic substances, aiming for
physicochemical, nutritional, and functional characteristics that closely resemble fresh
juice. The products processed with thermal treatments, pasteurization and sterilization,
have been questioned as they significantly alter the aroma and taste of foods due to the
formation of off-flavors. Therefore, consumer acceptance plays a crucial role in the food's
adoption into the diet and consequently in the commercial success of the product. Thus,
new processing routes and the implementation of emerging non-thermal technologies for
juice industrialization should prioritize maintaining quality and safety standards while
20
meeting consumer expectations regarding sensory characteristics such as taste, aroma,
and appearance. In this context, innovative non-thermal processing technologies have
been investigated with the aim of minimizing the degradation of nutritional compounds
and preserving sensory attributes similar to those of fresh products (Nonglait et al., 2022;
Pandiselvam et al., 2023; Zhu, Zhang, Mujumdar, & Liu, 2023).

In this case, PEF technology emerges as a promising alternative for the non-thermal
processing of fruit and vegetable juices. The application of PEF is facilitated by its
potential in deactivating pathogenic and spoilage microorganisms, endogenous enzymes,
and in preserving nutrients, physicochemical characteristics, and sensory attributes of the
beverages. By avoiding direct application of heat, PEF minimizes sensory alterations
compared to traditional thermal methods for processing fruit and vegetable juices. Studies
involving consumers dedicated to investigating the effects of PEF on the sensory quality
of treated fruit and vegetable juices are scarce.

Delso, Berzosa, et al. (2022) applied PEF (5 kV/cm, pulse width 40 µs, 120 Hz, 45
pulses, 63.4 kJ/kg) to red grapes to create a juice. This juice was treated with PEF (17.5
kV/cm, 16 pulses, pulse width 10 µs, 75 Hz, 173.6 kJ/kg) for microbiological
decontamination. Subsequently, a triangular test was conducted comparing the untreated
juice with the PEF-treated juice. The panel comprised 16 tasters (7 men and 9 women,
aged between 22 and 52 years). The tasters did not perceive a significant difference in the
flavor and aroma of the grape juice between the presented samples.

Lee et al. (2021) compared the characteristics of orange juice processed by thermal
treatments (95 ºC/30 s and 121 ºC/20 min) and PEF at 20 kV/cm (100 kJ/L and 150 kJ/L).
These treatments underwent a descriptive sensory analysis with seven trained panelists in
three repetitions using a 9 cm unstructured scale. The assessed attributes were aroma
similar to fresh juice, aroma similar to thermally treated juice, acidic taste, sweetness,
bitterness, and vegetal flavor. All attributes were evaluated in accordance with the
standards presented to the panelists. The PEF-treated juice samples showed no significant
differences compared to control samples for any of the seven attributes. The mild thermal
treatment showed no significant differences compared to the control sample; however,
the intense thermal treatment altered the aroma of fresh orange and thermally treated
orange as well as imparted a bitter taste.

Akdemir Evrendilek et al. (2020) evaluated the sensory characteristics of sour

cherry juice processed with two PEF treatments (24.7 kV/cm - 327 or 655 µs). The
analysis involved 25 trained panelists using a nine-point structured hedonic scale.
Evaluated attributes included color, aroma, sweetness, sour taste, and residual taste. Both
PEF treatments showed no significant differences compared to the untreated sample for
all five assessed attributes.

Wibowo et al. (2019) investigated the impact of PEF treatment (12.3 kV/cm - 132.5
kJ/L) on unclarified apple juice. The evaluation panel comprised 37 participants (7 men
and 27 women, aged between 23 and 58 years). The analysis was conducted using a 10-
centimeter scale. Evaluated attributes included sweet, sour, and bitter taste aspects. No
significant differences were observed in the bitter and sour taste attributes among the
different treatments. However, the panelists noted that samples subjected to PEF
treatment and high-pressure processing were perceived as the sweetest.

21
 In studies evaluating the sensory aspects of juices treated with PEF compared to
other technologies and thermal treatments, consumers were unable to differentiate
between PEF-treated samples and the untreated sample. This represents an opportunity
for the application of PEF technology in the processing of vegetable and fruit juices since
PEF is a technology that reduces deteriorative factors such as enzymes and
microorganisms while preserving the sensory characteristics of the products.

9. Application of PEF on vegetable and fruit juice industry

The PEF processing is a non-thermal and innovative technology based on

electroporation, which can be applied for microbial and enzymatic inactivation, recovery,
and preservation of bioactive compounds and pigments, among others. Therefore, PEF-
based processes in vegetable and fruit juices is an alternative to thermal processes
employed for stabilization, as this technology can result in kinetically and chemically
stable products without compromising bioactive compounds and sensory characteristics,
as presented in this review. Pasteurization is the most used thermal process in the
processing of vegetable and fruit juices, which not only alters the physicochemical
characteristics and content of bioactive compounds but also compromises nutritional
quality, promoting changes in the sensory characteristics of the products. In this context,
PEF technology emerges as an alternative to pasteurization in the juice industry.

For the application of the PEF in the juice industry, studies should be conducted to
determine efficient process variables concerning technical and economic viability. For
instance, the set of applied variables, such as field strength, frequency, pulse width, and
total treatment duration, should ensure complete inactivation of the target microorganism
for each specific product, and reduce the activity of deteriorating enzymes. Economically,
the PEF application should offset the costs of installation, maintenance, and energy
consumption of this technology, allowing large-scale production. Additionally, the price
must be affordable for the consumer.

Some studies investigate the feasibility of applying PEF on an industrial scale in

the vegetable and fruit juice sector. The main applications evaluated the PEF effect on
juice extraction yield. Bazhal, Lebovka, and Vorobiev (2001) studied the effects of PEF
on apple juice yield on an industrial scale. An increase in juice extraction yield after PEF
application was observed, and there was no significant difference in some quality aspects
compared to untreated juice, such as TSS, mineral content and phenolic compounds.
Turk, Billaud, Vorobiev, and Baron (2012) applied PEF for obtaining apple cider on a
pilot scale. An increase in extraction yield and improvement in the nutritional quality of
the beverage due to PEF treatment was verified. However, the content of phenolic
compounds was affected, and the product color was altered. Vignali et al. (2022)
conducted a feasibility study of different technologies used in apple juice processing with
the aim of reducing Escherichia coli by 5 log CFU/mL. The results indicated that PEF
showed higher energy efficiency (301.81 kJ/kg) compared to conventional thermal
treatment (397.54 kJ/kg), OH (371.16 kJ/kg), microwave heating (446.57 kJ/kg), and
HPP (339.94 kJ/kg). However, ultraviolet light radiation (26.24 kJ/kg), pulsed light
radiation (25.15 kJ/kg), and pressure change technology (26.28 kJ/kg) were more energy-
efficient when compared to PEF. The authors also highlighted that PEF technology, HPP,

22
 and ultraviolet light radiation already have industrial applications, while the other
technologies face scaling challenges.

Considering the data presented in this review, the innovative PEF technology has
demonstrated potential for application on an industrial scale in the processing of vegetable
and fruit juices. Some of the key factors for the chemical and kinetic stability of a juice
involve controlling enzymatic activity and the development of deteriorating
microorganisms. Thus, thermal treatments are conventional methods capable of
addressing these issues. However, the application of severe heat treatment may lead to
the loss of nutritional and sensory quality in juices. The installation cost and high energy
consumption pose challenges for the implementation of PEF. Despite these challenges,
this technology still has the potential to be scaled up and employed on an industrial scale,
resulting in a non-thermal processing method with lower energy expenditure and
achieving results similar to conventional thermal treatments.

10. Opportunities and challenges

The PEF technology emerges as a non-thermal food processing alternative.

However, this technique has several gaps that need to be studied, explored, and
elucidated. For instance, it is crucial to comprehend mechanisms related to enzymatic and
microbial inactivation, as well as intracellular compound extraction, among other factors.

In the specific context of vegetable and fruit juices, applying PEF to achieve safe
and kinetically and chemically stable products demands a detailed analysis of key process
variables. These variables encompass the field strength (kV/cm), frequency (Hz), pulse
width (µs), number of pulses, total treatment duration, and other factors that significantly
impact the final product's characteristics. Additionally, comprehending the complex
interactions between the intrinsic features of raw materials and process conditions is
essential.

In the context of vegetable and fruit juices, applying PEF to obtain stable products
needs to be studied to determine the main process variables (field strength, frequency,
pulse width, total treatment duration, among others) that impact the final product
characteristics. Furthermore, understanding the interactions between the intrinsic
characteristics of raw materials and process conditions becomes necessary.

The information presented previously is primary for implementing PEF technology

in vegetable and fruit juice processing into an industrial scale, providing not only safety
and chemical/kinetic stability to products but also data for advancing the development of
this non-thermal food processing alternative.

11. Conclusion

The innovative PEF technology is a promising alternative for processing vegetable

and fruit juices. In this review, the potential of PEF in reducing microbial load and
undesirable enzymatic activity without significant impacts on the physicochemical and
sensory characteristics of the products was examined. Additionally, an increase in the
23
availability of phenolic compounds, flavonoids, and anthocyanins, substances with
antioxidant, anti-inflammatory, anticancer properties, preventing chronic diseases, and
contributing to the appearance of the juices, was observed. Some application conditions
of PEF have proven that the technology has a low severity level by degrading little or
preserving ascorbic acid. However, future studies on the application of PEF technology
are necessary. Issues related to microbial and enzymatic inactivation, obtaining
intracellular compounds, and other aspects need to be investigated more deeply. There
are also recommendations for studies on PEF process design with low severity, aiming
for effective chemical, microbiological, and kinetic stabilization of vegetable and fruit
juices.

Acknowledgments
This research was funded by the São Paulo Research Foundation - FAPESP (grant
number # 2020/11255-3). Iuri Procopio Castro Brito thanks FAPESP for his master's
scholarship (grant number # 2023/04223-6). Eric Keven Silva thanks FAPESP (grant
number 2023/01876-9) for the Young Investigator Fellowship.

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34
35
Figure captions

Figure 1. Scheme of the electroporation phenomenon.

Figure 2. Systems of PEF application.

Eletroporation phenomenon

Eukaryotic cell

Cell membrane (lipid bilayer)

Reversible electroporation Irreversible electroporation

Normal membrane

Membrane during
application of PEF
PORE PORE PORE PORE

Membrane after
application of PEF
PORE PORE

36
 Systems of PEF application

Control system Pulse generator Oscilloscope

Untreated juice
Highlights
Continuous

- Pulsed electric field (PEF) is a promising non-thermal technology to vegetable and

fruit juices.
-
- PEF does not compromise the content of substances with functional and sensory
properties. Flow meter +

Pump
- PEF technology exhibited a low severity level by Treatment
degradingchamber
little or preserving
ascorbic acid. Treated juice

- PEF techniqueControl
appliedsystem
to juice processing has several gaps that need to be studied and
Pulse generator
elucidated. -
Batch

Treatment chamber
Oscilloscope

37
 Vegetables and
fruits Electroporation phenomena:
• Reversible
• Irreversible
Vegetable and
Spinach fruit juices

PEF
Strawberry technology

Orange PEF process variables:

• Field strength
• Frequency
• Pulse width
• Total treatment time
Other vegetables • Specific energy
and fruits

Table 1. Impacts of PEF treatment on vegetable and fruit juices concerning microbial
inactivation.

Treatment
Beverage Key findings References
conditions

Field strength:
19 kV/cm

Frequency: 1.25
kHz
PEF reduced the microbial load of YM from 5.5 to 1.2 (Jin e
Pulse width: 2
Orange juice log CFU/mL, and for TAMB, the reduction was less Aboelhaggag,
µs
than 1 log CFU/mL. 2022)
Treatment time:
181 µs

Specific energy:
-

38
 Field strength: 5
e 17.5 kV/cm

Frequency: 120
e 75 Hz

Pulse width: 10 - PEF showed a reduction greater than 3 log CFU/mL for
(Delso et al.,
Grape juice 40 µs molds, greater than 2 log CFU/mL for yeasts, and
2022)
approximately 1.0 log CFU/mL for TAMB.
Treatment time:
0.22 - 0.38 s

Specific energy:
63.4 e 173.6
kJ/L

Field strength:
10 - 21 kV/cm

Frequency: 235 -
588 Hz
Blend of
pineapple juice Pulse width: 2 PEF reduced the load of Escherichia coli and Listeria (Yildiz et al.,
with coconut µs innocua by 5.0 and 3.9 log CFU/mL, respectively. 2022)
milk
Treatment time:
-

Specific energy:
-

Field strength:
11.3 - 23.3
kV/cm

Frequency: 100 -
500 Hz
(Buitimea-
PEF promoted reductions of 0.92 to 2.12 log CFU/mL
Raspberry juice Pulse width: 25 Cantúa et al.,
for molds and 1.38 to 3.19 log CFU/mL for yeasts.
µs 2022)

Treatment time:
-

Specific energy:
19.7 - 168.4 kJ/L

Field strength:
20 - 40 kV/cm

Frequency: 1 Hz
PEF showed a reduction range of 5.891 to 5.924 log
(Kantala et al.,
Orange juice CFU/mL for Staphylococcus aureus and 5.876 to 5.949
Pulse width: 10 2022)
for Escherichia coli.
µs

Treatment time:
100 - 500 µs

39
 Specific energy:
-

Field strength: 5
kV/cm

Frequency: -

Pulse width: - PEF reduced the microbial load of TAMB by 0.27 log (Ahmed et al.,
Pineapple juice
CFU/mL 2022)
Treatment time:
20 min

Specific energy:
-

Field strength:
35 kV/cm

Frequency: 155
Hz

Pulse width: 2 PEF reduced approximately 1.21 log CFU/mL of (Yildiz et al.,
Strawberry juice
µs TAMB and 1.6 log CFU/mL of YM. 2021)

Treatment time:
27 μs

Specific energy:
-

Field strength:
21 - 34 kV/cm

Frequency: 500 -
1500 Hz
Apple, pepper, (Mendes-
Pulse width: 2 PEF reduced more than 5.0 log CFU/mL of
and passion fruit Oliveira et al.,
µs Escherichia coli and Salmonella Typhimurium.
juice 2021)
Treatment time:
72 - 217 μs

Specific energy:
-

Field strength: 9
kV/cm
Reduction of 0.37 log CFU/mL of Escherichia
Frequency: 1 (Ahmed et al.,
Wheatgrass juice coli/coliforms, 1.61 log CFU/mL of YM, and 1.99 log
kHz 2021)
CFU/mL of TAMB.
Pulse width: 80
µs

40
 Treatment time:
335 μs

Specific energy:
-

Field strength:
19 - 30 kV/cm

Frequency: 1.25
kHz
Reduction of 2.01 to 4.01 log CFU/mL for Escherichia
Pulse width: 2
Apple juice coli K-12, approximately 1 log CFU/mL for TAMB, (Jin et al., 2021)
µs
and 5.0 log CFU/mL for YM.
Treatment time:
181 μs

Specific energy:
-

Field strength:
15 - 30 kV/cm

Frequency: 200
Hz
Reduction of approximately 3 log CFU/mL for TAMB,
Cantaloupe Pulse width: 2
2 log CFU/mL for YM, and from 2.5 to 5.0 log (Li et al., 2021)
melon juice µs
CFU/mL for Saccharomyces cerevisiae, respectively.
Treatment time:
200 - 800 μs

Specific energy:
0.73 - 2.93 kJ/L

Field strength:
24.7 kV/cm

Frequency: 500
Hz Reduction of 0.60 to 1.26 log CFU/mL for Bacillus
cereus, 0.96 to 1.14 log CFU/mL for Pseudomonas
Sour cherry (Evrendilek et
Pulse width: 3 μs syringae subs. syringae, 1.04 to 1.19 log CFU/mL for
juice al., 2019)
Penicillium expansum, and 1.20 to 1.36 log CFU/mL
Treatment time: for Escherichia coli O157:H7, respectively.
327 e 655 μs

Specific energy:
-

Field strength:
11.9 kV/cm
Strawberry and Reduction of 5.59 and 3.62 log CFU/mL for the (Stübler et al.,
Frequency: -
kale juice microbial load of TAMB. 2020)
Pulse width: 20
μs

41
 Treatment time:
-

Specific energy:
120 kJ/kg

Field strength:
30 kV/cm

Frequency:
0.033 Hz
Reduction of 7.30 to 7.64 log CFU/mL for TAMB, 6.80
(Dziadek et al.,
Apple juice Pulse width: - to 7.00 log CFU/mL for molds, and 6.24 to 6.60 log
2019)
CFU/mL for yeasts, respectively.
Treatment time:
-

Specific energy:
-

Field strength:
35 kV/cm

Frequency: -

Pulse width: 2
Reduction of 2.90 log CFU/mL for TAMB and 3.69 log (Yildiz et al.,
Strawberry juice µs
CFU/mL for YM. 2019)
Treatment time:
27 μs

Specific energy:
-

MY = Molds and yeasts

TAMB = Total aerobic bacteria / Aerobic and mesophilic bacteria

Table 2. Effects of PEF treatment on vegetable and fruit juices for total acidity (TA), pH,
and total soluble solids (TSS).

Treatment
Beverage Key findings References
conditions

Field strength:
19 kV/cm (Jin e
There was no significant difference between the PEF
Orange juice Aboelhaggag
treatment and the control for these characteristics.
Frequency: 1.25 2022)
kHz

42
 Pulse width: 2
µs

Treatment time:
181 µs

Specific energy:
-

Field strength:
20 - 40 kV/cm

Frequency: 1 Hz

Pulse width: 10
There was no significant difference between the PEF (Kantala et al.,
Orange juice µs
treatment and the control for pH and TSS. 2022)
Treatment time:
100 - 500 µs

Specific energy:
-

Field strength:
11.3 - 23.3
kV/cm

Frequency: 100
- 500 Hz
PEF increased the pH from 3.31 to 3.36 to 3.38, (Buitimea-
Pulse width: 25
Raspberry juice increased the TSS from 7.5 to 8.5 ºBrix, and decreased Cantúa et al.,
µs
the TA from 1.00 to 0.9% citric acid. 2022)
Treatment time:
-

Specific energy:
19.7 - 168.4
kJ/L

Field strength:
1.01 kV/cm

Frequency: 2 Hz

Pulse width: 80
PEF decreased the pH from 7.03 to 6.73 and increased (Atencio et al.,
Pumpkin juice - 81 µs
the TSS from 2.54 to 2.80 ºBrix. 2022)
Treatment time:
-

Specific energy:
9.0 kJ/kg

PEF treatment did not induce changes in TA, except for

Chokeberry Field strength: (Oziembłowski
one juice sample, which exhibited an increase in this
juice 20 kV/cm et al., 2022)
characteristic.

43
 Frequency:
0.066 Hz

Pulse width:
0.17 µs

Treatment time:
-

Specific energy:
-

Field strength: 5
kV/cm

Frequency: -

Pulse width: - PEF increased the pH from 4.14 to 4.22 and reduced the (Ahmed et al.,
Pineapple juice
TSS from 12.06 to 11.03 ºBrix. 2022)
Treatment time:
20 min

Specific energy:
-

Field strength: 0
- 26.7 kV/cm

Frequency: 0 -
500 Hz

Sour cherry Pulse width: 3 There was no significant difference between the PEF (Evrendilek et
juice µs treatment and the control for these characteristics. al., 2021)

Treatment time:
0 - 873.1 μs

Specific energy:
0 - 0.0341 J/L

Field strength:
35 kV/cm

Frequency: 155
Hz

Strawberry Pulse width: 2 There was no significant difference between the PEF (Yildiz et al.,
juice µs treatment and the control for these characteristics. 2021)

Treatment time:
27 μs

Specific energy:
-

44
 Field strength:
21 - 34 kV/cm

Frequency: 500
- 1500 Hz
Apple, pepper, (Mendes-
Pulse width: 2 There was no significant difference between the PEF
and passion Oliveira et al.,
µs treatment and the control for these characteristics.
fruit juice 2021)
Treatment time:
72 - 217 μs

Specific energy:
-

Field strength: 9
kV/cm

Frequency: 1
kHz

Pulse width: 80 There was no significant difference between the PEF (Manzoor et al.,
Spinach juice
µs treatment and the control for TSS. 2021)

Treatment time:
335 μs

Specific energy:
-

Field strength:
19 - 30 kV/cm

Frequency: 1.25
kHz

Pulse width: 2 There was no significant difference between the PEF

Apple juice (Jin et al., 2021)
µs treatment and the control for these characteristics.

Treatment time:
181 μs

Specific energy:
-

Field strength:
15 - 30 kV/cm

Frequency: 200
Hz
Cantaloupe There was no significant difference between the PEF
(Li et al., 2021)
melon juice treatment and the control for pH and TSS.
Pulse width: 2
µs

Treatment time:
200 - 800 μs

45
 Specific energy:
0,73 – 2,93 kJ/L

Field strength: 0
- 14 kV/cm

Frequency: 1
kHz
PEF decreased the pH from 2.99 to 2.92 and reduced the
(Rahaman et al.,
Apricot juice Pulse width: - TSS from 4.01 to 3.80 ºBrix with increased treatment
2020)
intensity.
Treatment time:
500 μs

Specific energy:
-

Field strength:
24.7 kV/cm

Frequency: 500
Hz

Sour cherry Pulse width: 3 There was no significant difference between the PEF (Evrendilek et
juice μs treatment and the control for these characteristics. al., 2020)

Treatment time:
327 e 655 μs

Specific energy:
-

Field strength:
12.3 e 12.5
kV/cm

Frequency: 62 e
94 Hz PEF increased the pH from 3.32 to 3.45 and reduced the
TSS from 13.08 to 12.78 ºBrix in the less intense
Cloudy apple Pulse width: 2 treatment. However, the more intense treatment showed (Wibowo et al.,
juice μs no significant difference. Meanwhile, the TA did not 2019)
show a significant difference between the PEF treatment
Treatment time: and the control.
-

Specific energy:
76.4 e 132.5
kJ/L

46
 Treatment
Beverage Key findings References
conditions

Field strength:
20 - 40 kV/cm

Frequency: 1 Hz

Pulse width: 10
There was no significant difference in color variation (Kantala et al.,
µs
between the PEF treatment and the control. 2022)
Orange juice
Treatment time:
100 - 500 µs

Specific energy:
-

Field strength: 5
kV/cm

Frequency: -
Among the evaluated treatments (microwaves, vacuum
Pineapple Pulse width: - (Ahmed et al.
evaporation, pasteurization, PEF, and ultrasonication), PEF
juice 2022)
showed the most significant color alteration (6.19).
Treatment time:
20 min

Specific energy:
-

Field strength: 9
kV/cm

Frequency: 1
kHz
Among the evaluated treatments (ultrasound, PEF, and
Spinach Pulse width: 80 (Manzoor et al.,
ultrasound + PEF), the PEF treatment showed the least color
juice µs 2021)
variation (1.73).
Treatment time:
335 μs

Specific energy:
-

Field strength:
19 - 30 kV/cm
Among the treatments performed with pulsed light and PEF,
the intensity of 19 kV/cm of PEF resulted in less intense color
Frequency: (Jin et al.,
Apple juice changes (0.62) compared to the intensity of 23 kV/cm (1.47).
1.250 kHz 2021)
Thus, 0.62 represented the smallest color variation observed
among the treatments.
Pulse width: 2
µs

47
 Treatment time:
181 μs

Specific energy:
-

Field strength: 0
- 14 kV/cm

Frequency: 1
kHz
The PEF with a field intensity of 7 kV/cm showed a greater
Apricot (Rahaman et
Pulse width: - color change (0.69) than the juice treated with 14 kV/cm
juice al., 2020)
(0.56).
Treatment time:
500 μs

Specific energy:
-

Field strength:
24.7 kV/cm

Frequency: 500
Hz
Among the evaluated treatments (ozone, PEF, and
Sour cherry Pulse width: 3 (Evrendilek et
ultrasonication), the PEF treatments showed the lowest color
juice μs al., 2020)
variation (0.88 and 0.89).
Treatment time:
327 e 655 μs

Specific energy:
-

Table 3. Impact of PEF treatment on the color of vegetable and fruit juices.

Table 4. Effects of PEF on the inactivation of pectin methylesterase (PME), polyphenol

oxidase (PPO), and peroxidase (POD) in vegetable and fruit juices.

Treatment
Beverage Key findings Referências
conditions

Field strength:
11.3 - 23.3
kV/cm
(Buitimea-
Raspberry PEF treatments with 168.4, 79.7, and 113.8 kJ/L led to
Frequency: 100 - Cantúa et al.,
juice inhibitions of 19%, 13%, and 13% in PME activity.
500 Hz 2022)

Pulse width: 25
µs

48
 Treatment time:
-

Specific energy:
19.7 - 168.4 kJ/L

Field strength: 9
kV/cm

Frequency: 1
kHz

Spinach Pulse width: 80 PEF reduced the activity of PPO and POD by 43.25% and (Manzoor et al.,
juice µs 44.00%, respectively. 2021)

Treatment time:
335 μs

Specific energy:
-

Field strength: 9
kV/cm

Frequency: 1
kHz

Wheatgrass Pulse width: 80 PEF reduced the activity of PPO and POD by 38.70% and (Ahmed et al.,
juice µs 35.63%, respectively. 2021)

Treatment time:
335 μs

Specific energy:
-

Field strength:
12.3 e 12.5
kV/cm

Frequency: 62 e
94 Hz Reduction greater than 90% and 36% in PPO activity and
greater than 90% and 49% in POD activity were observed
Cloudy Pulse width: 2 for PEF treatments of 132.5 and 76.4 kJ/L, respectively. (Wibowo et al.,
apple juice μs The PEF treatment at 132.5 kJ/L led to complete 2019)
inactivation of PME, while the 76.4 kJ/L dose reduced
Treatment time: activity by 50%.
-

Specific energy:
76.4 e 132.5
kJ/L

PEF reduced PPO activity from 100% to 72.73%, 94.81%,

Field strength: (Tian et al.,
Apple juice and 92.21% for treatments of 15, 25, and 35 kV/cm,
15 - 30 kV/cm 2018)
respectively.

49
 Frequency: 200
Hz

Pulse width: 2
µs

Treatment time:
200 - 800 μs

Specific energy:
0.73 – 2.93 kJ/L

Table 5. Effects of PEF treatment on the phenolic compounds of vegetable and fruit
juices.

Treatment
Beverage Key findings References
conditions

Field strength: 19
kV/cm

Frequency: 1.25
kHz
There was no significant difference between the PEF (Jin e
Orange juice treatment and the control for the content of these Aboelhaggag,
Pulse width: 2 µs
substances. 2022)
Treatment time:
181 µs

Specific energy: -

Field strength:
11.3 - 23.3 kV/cm

Frequency: 100 -
500 Hz
PEF treatment increased the phenolic compounds
(Buitimea-Cantúa
content by 35 to 74% according to the increase in the
Raspberry Pulse width: 25 µs et al., 2022)
intensity and frequency of the electric field.
juice
Treatment time: -

Specific energy:
19.7 - 168.4 kJ/L

Field strength: 20
kV/cm There was no significant difference between the PEF
Chokeberry (Oziembłowski et
treatment and the control for the content of these
juice al., 2022)
Frequency: 0.066 substances.
Hz

50
 Pulse width: 0.17
µs

Treatment time: -

Specific energy: -

Field strength: 5
kV/cm

Frequency: -
Pineapple PEF reduced the content of phenolic compounds by (Ahmed et al.,
Pulse width: -
juice 5.6% compared to the control sample. 2022)
Treatment time:
20 min

Specific energy: -

Field strength: 0 -
26.7 kV/cm

Frequency: 0 -
500 Hz
There was no significant difference between the PEF
Sour cherry (Evrendilek et al.,
Pulse width: 3 µs treatment and the control for the content of these
juice 2021)
substances.
Treatment time: 0
- 873.1 μs

Specific energy: 0
- 0.0341 J/L

Field strength: 35
kV/cm

Frequency: 155
Hz
There was no significant difference between the PEF
Strawberry (Yildiz et al.,
treatment and the control for the content of these
juice Pulse width: 2 µs 2021)
substances.
Treatment time:
27 μs

Specific energy: -

Field strength: 9
kV/cm

Frequency: 1 kHz
Spinach PEF increased the content of phenolic compounds by (Manzoor et al.,
Pulse width: 80 µs
juice 8.49% compared to the control sample. 2021)
Treatment time:
335 μs

Specific energy: -

51
 Field strength: 9
kV/cm

Frequency: 1 kHz
Wheatgrass PEF increased the content of phenolic compounds by (Ahmed et al.,
Pulse width: 80 µs
juice 5.35% compared to the control sample. 2021)
Treatment time:
335 μs

Specific energy: -

Field strength: 19
- 30 kV/cm

Frequency: 1.250
kHz The less intense PEF treatment (19 kV/cm) reduced the
content of phenolic compounds by 3.39%. However,
Apple juice (Jin et al., 2021)
Pulse width: 2 µs the treatment at 23 kV/cm did not significantly alter the
content of these substances.
Treatment time:
181 μs

Specific energy: -

Field strength: 0 -
14 kV/cm

Frequency: 1 kHz
PEF increased the content of phenolic compounds by (Rahaman et al.,
Apricot juice Pulse width: -
3.40% and 8.25% compared to the control sample. 2020)
Treatment time:
500 μs

Specific energy: -

Field strength:
24.7 kV/cm

Frequency: 500
Hz
There was no significant difference between the PEF
Sour cherry (Evrendilek et al.,
treatment and the control for the content of these
juice Pulse width: 3 μs 2019)
substances.
Treatment time:
327 e 655 μs

Specific energy: -

Field strength: 30
kV/cm There was no significant difference between the PEF
(Dziadek et al.,
Apple juice treatment and the control for the content of these
2019)
Frequency: 0.033 substances.
Hz

52
 Pulse width: -

Treatment time: -

Specific energy: -

Declaration of interests

☒ The authors declare that they have no known competing financial interests or
personal relationships that could have appeared to influence the work reported in this
paper.

☐The authors declare the following financial interests/personal relationships which may be
considered as potential competing interests:

Eric Keven Silva

School of Food Engineering

UNICAMP - University of Campinas

Rua Monteiro Lobato, 80

13083-862 Campinas, SP – Brazil

E-mail address: engerickeven@gmail.com

53