FPP - Pulse Applications I - 7-12-22

Pulse Power Industrial Applications
- Application of Pulsed Electric Fields,

electroporation “Biological effects”
7/12/22 FPP – Pulse applications I - Luís Redondo 1

PP industrial applications – Summary
Biological effects:
Where moderate pulse - food processing (mass transfer & inactivation)
energies, repetitive - plasma decontamination
- medical treatment (e.g. cancer)
pulses, efficient,
- crop growth
portable, Streamer discharges in gases:
reliable and - exhaust gas treatment
cost effective - ozone generation
modulators are Escreva uma inequação
Discharges liquid or aqui.
liquid-mixture:
needed. - water treatment
- engine ignition
Material processing:
Loads: - implantation & deposition (surface change)
R, liquids - magnetic forming, welding & cutting
RC, gases, plasmas - nano particles synthesizing
RL, inductor, transformer - concrete recycling
- laser (material ablation & surface annealing)
In average: up to kHz, µs, Accelerators & physics:
100’s A, 10’s kV. - Thyratron replacement
- Breakdown studies

PP industrial applications – Biological effects
Application of electric fields to biological cells - Cell
Intact cell: selective membrane permeability
• cell plasma membrane tightly

controls the traffic of molecules
between the cell and its
environment
Escreva uma equação aqui.
• through ion channels/pumps
and endo-/exocytosis
• from electrical point of view the

membrane is a poor
conductor (or capacitor)
Kotnik T, Rems L, Tarek M, Miklavčič D.

Membrane Electroporation and Electropermeabilization:
Mechanisms and Models.
Annu. Rev. Biophys. 48: 63-91, 2019.

Application of electric fields to biological cells - Electroporation
The exposition of biological lipid

bilayers to an external electric
field of enough intensity causes
an increase of their permeability,
i.e. the pores formation in cell
membrane, in order to distribute
the electrical charges of the cells
Escreva umainequação
the two aqui.
sides of the
phospholipidic bilayer. This effect
called electroporation
• induced membrane potential difference ≈ 500 mV over membrane

thickness ≈ 5 nm
• establishing huge electric field Electrical breakdown in air
within the membrane occurs at E > 3 MV/m
500 mV
= 100 MV/m
5 nm

Ø Charging of membrane [ t < 1μs];

Ø Stress on the membrane and pore
formation;
Ø Expansion of pore radii and
aggregation of different pores [ t
>100μs];
Ø Pore resealing and memory effects
[seconds to hours]

Aqueous pore formation theory

Aqueous pore formation theory

Ec – Critical electric field

Um
Transmembrane voltage threshold

θ
(0.5V to 1V)
Cell
?
>
𝑈2 = 1.5 𝑟 𝐸 𝑐𝑜𝑠𝜃 (1 − 𝑒 @ )

Application of electric fields to biological cells – Electroporation, PEF
𝐸
U
+
𝑈
𝐸 = [𝑘𝑉 ⁄𝑐𝑚]
𝑑
Electroporation can be achieved, also, by the intermittent delivery of short pulses of

high voltage (kV) to a biological medium located between two electrodes (Pulsed
Electric Fields, PEF).
PEF have the advantage of reducing the thermal load applied to the material,
making it possible to apply intense fields in the order of kV/cm to biological
samples, but without thermal load.

Exposure to
electric field irreversible Cell death Tissue ablation
+ E - Pasteurisation
reversible Introducing small Electrochemotherapy
molecules tumor treatment
Escreva Extraction
uma equaçãoof aqui. Biotechnology
molecules
Introducing large
DNA vaccination
molecules
Gene therapy
Cell fusion
Protein insertion in
*synonyms:
electropermeabilization, the plasma membrane
nucleofection, PEF treatment,
nanoporation, PFA,…

Application of electric fields to biological cells – PEF
Change of scale:
Ø Target: cells (bacteria, vegetable, animal);
Ø Effect: electroporation – field effect;
Ø Treatment time: microseconds; Escreva uma equação aqui.
Ø Processing time: < 1 second;
Ø Energy consumption: low (compressed, defined)
Ø Treatment intensity: high (kilovolts)

Application of electric fields to biological cells - PEFn
𝐶 − Capacitance R − Resistance
S − Switch 𝐼 − Current [A]

𝑈LI − Voltage [V] 𝑡RS − pulse width [µ𝑠]
1 M
𝐸I = 𝐶 𝑈LI [𝐽] ≫ 𝐸O = 𝑈LI 𝐼 𝑡RS [𝐽]
2
𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑜𝑟 𝐸𝑛𝑒𝑟𝑔𝑦 𝑃𝑢𝑙𝑠𝑒 𝐸𝑛𝑒𝑟𝑔𝑦

Electroporation applications
Medical and Biomedical
Djokic et al. 2018

Reversible and Irreversible
electroporation
Environmental
Irreversible electroporation
Food Industry
Reversible and Irreversible
electroporation

Medical applications
1. Electrochemotherapy
selectivity of ECT10 weeks after ECT

2. Gene electrotransfer for gene therapy and DNA vaccination
3. Soft tissue/tumor ablation with irreversible electroporation
Miklavcic et al, Biomed Eng Online, 2010

Yarmush et al, Annu Rev Biomed Eng, 2014

Environmental applications
1. Biogas production
In many wastewater treatment plants (WWTPs), a

community of anaerobic microorganisms degrades
organic matter in waste biosolids and converts the
energy value to methane gas (CH4).

Higher biomass destruction and conversion to CH4
offers important economic benefits to a WWTP, which
should encourage more facilities to adopt anaerobic
digestion, methane recovery, and use of the energy
value of CH4.
With PEF pretreated solids are made more
bioavailable, they are more completely
hydrolyzed, which leads to increased
solids destruction and lower solids-
disposal costs.

Food applications
Mass Transfer / Extraction Food Pasteurization

Electroporation of eukaryote cells Microbial Inactivation
Electric Field strength <5 kV/cm Electric Field strength >15 kV/cm
Low energy (<10kJ/kg) High energy (>50 kJ/kg)

Food applications
PEF for Mass transfer

Pre-treatment to optimize an extraction process.
Ø Extraction of intracellular compounds

Ø NFC fruit juices (apple, carrot)
Ø Sugar from beets
Ø Pigments and phenolic components
(winemaking, plant extracts)
Ø Removing water
Ø Drying fruits and vegetables
Ø Impregnation
Ø Osmotic dehydration
Ø Salting (ham)
Ø Brining (cooked ham)

Food applications
PEF for Food Pasteurization

Microbial Inactivation
Objectives Thermal Pasteurization
Ø Biologically safe product Ø Compromise between safety and

quality.
Ø High quality – fresh-like product
Ø Decrease of quality (color, flavor,
Ø Reasonable shelf-life degradation of components)
Emerging
Technologies
(industry)

Food applications - formulas
Electric field Specific energy
𝑈 𝐸s 𝑁 𝑈 𝐼 𝜏 𝑁
𝐸 = [𝑘𝑉 ⁄𝑐𝑚] 𝐸m =
𝑚
=
𝑚
[𝑘𝐽/𝑘𝑔]
𝑑
Electric current Temperature increase
𝑈 𝑑
𝐼= 𝑅=𝜌 Ω 𝐸y
𝑅 𝐴m ∆𝑇 = , [℃]
𝐶O
1
𝜌 = [Ω r m]
𝜎
CP, specific heat, J/Kg.K,
where 1 cal = 4,186 J
𝑈𝐴m 𝜎
𝐼= [𝐴]
𝑑

Food applications - Examples
EnergyPulse Systems Equipment – Treatment chambers

EnergyPulse Systems Industrial PEF process in-line
Double co-field
flow chamber

EnergyPulse Systems Industrial PEF applications
Apple juice Potato Chips

Ø Yield increase Ø Reduction of frying time
Ø Reduction of enzymes Ø Smoother cutting
Ø Reduction of pressing time Ø Acrylamide reduction
Ø “Cold” microbial inactivation
Wine Olive oil

Ø Faster release of polyphenols Ø Reduction of malaxation temperature
Ø Colour increase Ø Reduction of pomace fat
Ø Juice yield increase
Microalgae Tomato
Ø Control of contaminants Ø Steam temperature reduction for peeling
Ø Lipids extraction Ø 20% energy savings

PEF for Potatoes

u Chips and French fries
Ø Reduction of turgor pressure
Ø Reduction of acrylamide content on potato
chips.
Ø Improve of crispiness and color attributes.

PEF for Apple Juice

u Yield increase
• one of the largest producer of
apple juice & juice concentrates.
Ø Not from concentrate, NFC, juice
(using belt presses), average 2.5
% yield increase.
u Enzymes reduction
Ø Concentrate juice (with Bucher
presses), reduction of 50 %
enzymes for the same yield.

PEF for wine
u Work in about 20 wineries in Portugal

and Spain, with red and white grapes
showed:
Red grapes:
• Faster release of polyphenols and color
intensity, which accelerates the
maceration process up to three times
White grapes:
• Higher efficiency of the first pressure up
to 10% more juice

PEF for olive oil
u Olive oil cellar

Ø Increase of olive oil yield, up to 5%
more
Ø Reduction of pomace fat up to 15 , for
the same malaxation time.

PEF for peeling Tomato
Ø Combining technologies –
PEF and Heat
Ø Saving Energy – up to 20%
reduction on vapor pressure at
industrial level
Ø Improving circular economy –
waste reduction

Up Ip E f ton P T
Teste # Obs.
(kV) (A) (kV/cm) (Hz) (µ
µs) (W) (ºC)

0 - - - - - - 22 -
1 4 70 1 100 25 700 23 30 min (início 10h30)
2 4 70 1 100 50 1400 23,5 30 min
3 4 75 1 50 100 1500 24 30 min
Food applications - Examples 4

5
6
6
115
118
1,5
1,5
50
20
50
100
1725
1416
24,5
26
30 min
30 min
6 5,5 110 1,375 50 50 1513 26,5 90 min, hora de almoço (13h às 14h30)
7 6 120 1,5 20 100 1440 26,5 30 min
PEF for microalgae

8 6 120 1,5 50 50 1800 26,7 60 min
9 4 80 1 50 105 1680 25 30 min (fim às 16h30)
• Foram recolhidas amostras a cada teste (a cada 30 min), que foram observadas ao microscópio (OM). Adicionalmente, para
foram recolhidas mais amostras para melhor acompanhar o impacto do campo eléctrico na cultura.
• Posteriormente (12Mai30), as amostras mais relevantes foram sujeitas a análises de sobrenadante, para identificação e qua
pigmentos (representativos do meio intracelular).
• Nos dias após o ensaio, a cultura foi acompanhada com OM diária, para analisar a evolução da população de rotíferos, be
u Control of contaminants Responsáveis microalgas, após a aplicação do campo eléctrico.
Científico – Luís Costa e Vanessa Geraldes
Ø Microalgae, Chlorophyta, contaminated with Operacional – Jaime Páramo e Duarte Rego
Relatório – Duarte Rego
4% of protozoa, namely rotifers.
Resultados t0 (amostra inicial):
A cultura apresentava uma população variada de microalgas, bem como diversos contaminantes, maioritariamente rotíferos,
fungos, bactérias e ciliados, consequência do período de contaminação forçada anterior ao ensaio.
Dos contaminantes, a população de rotíferos representava perto de 4% da cultura, sendo a presença de ovos de rotíferos quase
2.7 m3 volume photobioreactor, 6 hours treatment
time,
1 euro is the estimated cost for complete washing-
out the protozoa after 6 days of treatment
application.
Figura 2 – Cultura do PBR_B1 no tempo 0 (t0).
Efeito do Campo Eléctrico na população de rotíferos:
Teste # Observações Fotografia
A cultura (microalgas e rotíferos) manteve a sua aparência
7/12/22 FPP – Pulse applications I - t1Luís Redondo geral inicial. Em relação à população de rotíferos, esta
apresentava pouca actividade (vídeo P1090868_t1), o que
28
não é frequente.
PEF for Juice preservation
u Cloudy Apple Juice
Feedin
g tank Ø Pre-heating 40 oC
Ø Monopolar pulses of 8 µs
Ø > log5 reduction with 100 kJ/kg

PEF for Juice Preservation

Bipolar pulses effect
Ø No pre-heating
Ø Monopolar pulses of 8 µs
Ø Bipolar pulses 2 x 4 µs
Ø Bipolar shows higher inactivation effect for E.
coli
Ø Lower final temperature with bipolar
Ø Bipolar beneficial for electrodes corrosion

FPP - Pulse Applications I - 7-12-22

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Pulse Power Industrial Applications

- Application of Pulsed Electric Fields,

7/12/22 FPP – Pulse applications I - Luís Redondo 1

7/12/22 FPP – Pulse applications I - Luís Redondo 2

Intact cell: selective membrane permeability

• cell plasma membrane tightly

• from electrical point of view the

Kotnik T, Rems L, Tarek M, Miklavčič D.

7/12/22 FPP – Pulse applications I - Luís Redondo 3

The exposition of biological lipid

• induced membrane potential difference ≈ 500 mV over membrane

7/12/22 FPP – Pulse applications I - Luís Redondo 4

Ø Charging of membrane [ t < 1μs];

7/12/22 FPP – Pulse applications I - Luís Redondo 5

Aqueous pore formation theory

Escreva uma equação aqui.

7/12/22 FPP – Pulse applications I - Luís Redondo 6

Aqueous pore formation theory

Escreva uma equação aqui.

7/12/22 FPP – Pulse applications I - Luís Redondo 7

Escreva uma equação aqui.

Ec – Critical electric field

Transmembrane voltage threshold

7/12/22 FPP – Pulse applications I - Luís Redondo 8

Electroporation can be achieved, also, by the intermittent delivery of short pulses of

7/12/22 FPP – Pulse applications I - Luís Redondo 9

7/12/22 FPP – Pulse applications I - Luís Redondo 10

7/12/22 FPP – Pulse applications I - Luís Redondo 11

S − Switch 𝐼 − Current [A]

7/12/22 FPP – Pulse applications I - Luís Redondo 12

Djokic et al. 2018

7/12/22 FPP – Pulse applications I - Luís Redondo 13

selectivity of ECT10 weeks after ECT

Escreva uma equação aqui.

3. Soft tissue/tumor ablation with irreversible electroporation

Miklavcic et al, Biomed Eng Online, 2010

7/12/22 FPP – Pulse applications I - Luís Redondo 14

In many wastewater treatment plants (WWTPs), a

Escreva uma equação aqui.

7/12/22 FPP – Pulse applications I - Luís Redondo 15

Mass Transfer / Extraction Food Pasteurization

7/12/22 FPP – Pulse applications I - Luís Redondo 16

PEF for Mass transfer

Ø Extraction of intracellular compounds

5/01/22 FPP – Pulse applications I - Luís Redondo 17

PEF for Food Pasteurization

Objectives Thermal Pasteurization

Ø Biologically safe product Ø Compromise between safety and

7/12/22 FPP – Pulse applications I - Luís Redondo 18

Electric current Temperature increase

7/12/22 FPP – Pulse applications I - Luís Redondo 19

EnergyPulse Systems Equipment – Treatment chambers

7/12/22 FPP – Pulse applications I - Luís Redondo 20

EnergyPulse Systems Industrial PEF process in-line

7/12/22 FPP – Pulse applications I - Luís Redondo 21

EnergyPulse Systems Industrial PEF applications

Apple juice Potato Chips

Wine Olive oil

7/12/22 FPP – Pulse applications I - Luís Redondo 22

PEF for Potatoes

7/12/22 FPP – Pulse applications I - Luís Redondo 23

PEF for Apple Juice